Cancer antigens

Abstract

The present invention relates to tumor or cancer antigens, in particular amino acid sequences and nucleic acid sequences, useful in the immunotherapy of cancer. In particular, the present invention relates to an artificial nucleic acid, preferably RNA, comprising at least one coding sequence encoding at least one tumor antigen or cancer antigen, said antigen comprising or consisting of at least one antigen peptide selected from or derived from a small open reading frame (smORF)-encoded peptide or protein, or a fragment or variant thereof, selected from or derived from long-chain non-coding RNA (lncRNA), and / or at least one antigenic peptide selected from or derived from a peptide or protein of a tumor neoantigen, or a fragment or variant thereof. Also provided is a pharmaceutical composition comprising the artificial nucleic acid, preferably a pharmaceutical composition formulated in a lipid-based carrier. Also provided are methods of treating or preventing a disorder, disease or condition, and medical uses, in particular for treating or preventing cancer, such as NSCLC, HNSCC or melanoma.

Description

[0001] introduction:

[0002] Cancer is a major global health problem and one of the leading causes of death worldwide. Traditional cancer treatments, such as surgery, chemotherapy, and radiation therapy, have limited efficacy and can cause significant side effects. Immunotherapy, which involves harnessing the power of the immune system to target cancer cells, has emerged as a promising new approach to cancer treatment. Cancer immunotherapy utilizes the immune system to treat cancer. Immunotherapy leverages the fact that cancer cells often have molecules called tumor antigens on their surface that can be detected by the immune system; these molecules are typically proteins or other macromolecules. Promising candidates for highly specific immunotherapies are neoantigens or tumor-associated antigens.

[0003] In recent years, nucleic acids, such as RNA, have emerged as promising therapeutic tools for treating cancer and other diseases because they can be used to guide the production of proteins that can inhibit or otherwise reduce the growth or survival of cancer cells. Advantages of using RNA include transient expression and non-conversion properties—RNA does not need to enter the cell nucleus to be expressed and cannot integrate into the host genome, thus eliminating the risk of tumorigenesis. Therefore, nucleic acids, such as RNA, may represent a promising class of molecules for providing information on the expression of tumor antigens such as neoantigens or tumor-associated antigens.

[0004] Therefore, the object of the present invention is to provide novel antigens or tumor-associated antigens for cancer immunotherapy, particularly novel antigens or tumor-associated antigens provided by nucleic acid constructs.

[0005] The aforementioned objectives are achieved through the following description and the appended claims. In particular, the objectives of the invention are achieved by providing novel tumor antigens, nucleic acids encoding such tumor antigens, and the use of these tumor antigens / nucleic acids in cancers of therapeutic subjects such as NSCLC, HNSCC, or melanoma. Specifically, tumor antigens unexpectedly derived from small open reading frames of long non-coding RNAs (as the term suggests, these RNAs have long been considered non-coding). Summary of the Invention

[0006] This invention relates particularly to novel tumor or cancer antigens, especially amino acid and nucleic acid sequences, for use in cancer immunotherapy. Specifically, the invention relates to artificial nucleic acids, preferably RNA, comprising at least one coding sequence encoding at least one tumor or cancer antigen, said antigen comprising or constituting at least one antigenic peptide selected from or derived from peptides or proteins encoded by small open reading frames (smORFs) of lncRNA, or fragments or variants thereof; and / or at least one antigenic peptide selected from or derived from peptides or proteins, or fragments or variants thereof, of tumor neoantigens. Pharmaceutical compositions comprising said artificial nucleic acids are also provided, preferably pharmaceutical compositions formulated in a lipid-based carrier. Methods for treating or preventing disorders, diseases, or conditions, and their medical uses are also provided, particularly for cancers such as NSCLC, HNSCC, or melanoma.

[0007] As illustrated herein, this invention is particularly based on the unexpected discovery that peptides encoded by small open reading frames (smORFs) of long non-coding RNAs (lncRNAs) can serve as effective tumor-associated antigens. Furthermore, novel antigens have been discovered that can serve as effective tumor-specific antigens. These tumor antigens can be provided via artificial nucleic acids, such as mRNA, for the treatment of cancers such as NSCLC, HNSCC, or melanoma.

[0008] Therefore, in a first aspect, the present invention provides an artificial nucleic acid comprising at least one coding sequence encoding at least one tumor antigen or cancer antigen, said antigen comprising (a) at least one antigenic peptide selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA); and / or (b) at least one antigenic peptide selected from or derived from peptides or proteins of tumor neoantigens.

[0009] In a preferred embodiment, at least one antigenic peptide is selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA), or immunogenic fragments or variants thereof, wherein the lncRNA is selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25.

[0010] In other preferred embodiments, at least one antigenic peptide is selected from peptides or proteins of tumor neoantigens, said tumor neoantigen being selected from NRAS, TYW1B, ECPAS, MAP2K1, TOMM22, GLB1, MAGE-A3, ATAD2, BRAF, EGFR, or TP53, or immunogenic fragments or variants thereof.

[0011] In a second aspect, the present invention provides a pharmaceutical composition comprising at least one artificial nucleic acid as defined in the first aspect. The artificial nucleic acid, preferably RNA, can be formulated in an LNP.

[0012] In a third aspect, the present invention provides tumor antigens or compositional antigens.

[0013] In a fourth aspect, the present invention provides antibodies, T cells, or TCRs that have been generated against any of at least one nucleic acid encoding a tumor antigen of the present invention, or stimulated using any of at least one nucleic acid encoding a tumor antigen of the present invention.

[0014] In a fifth aspect, the present invention provides a combination comprising at least two or more treatment modalities of any of the foregoing aspects.

[0015] In a sixth aspect, the present invention provides a kit or kit comprising at least one artificial nucleic acid of the first aspect; at least one pharmaceutical composition of the second aspect; at least one tumor antigen or composition of the third aspect; and / or at least one antibody, T cell or TCR of the fourth aspect.

[0016] In other respects, the present invention provides methods for treating or preventing diseases, disorders, or conditions, as well as medical uses of artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions, antibodies, T cells or TCRs, or kits or kits. Preferably, the disease, disorder, or condition is cancer, or any cancer-related disease, disorder, or condition, such as NSCLC, HNSCC, or melanoma.

[0017] definition

[0018] For clarity and readability, the following definitions are provided. Any technical features mentioned for these definitions can be found in each embodiment of the invention. Further definitions and explanations are provided in the context of these embodiments.

[0019] In the context of numbers, percentages should be understood as totals relative to the corresponding terms. In other cases, unless the context otherwise specifies, percentages should be understood as weight percentages (wt.-%).

[0020] about: The term "about" is used when the determinants or values ​​do not need to be exactly the same, i.e., 100% identical, and when, for example, the corresponding values ​​or determinants may differ by 1% to 10%. Preferably, "about" means that the determinants or values ​​may differ by + / -1%, + / -2%, + / -3%, + / -4%, + / -5%, + / -6%, + / -7%, + / -8%, + / -9%, or + / -10%.

[0021] Antigenic peptide: As used herein, the terms “antigenic peptide” or “immunogenic peptide” refer to a peptide derived from an antigenic or immunogenic protein that stimulates the body’s adaptive or cellular immune system to provide an adaptive or cellular immune response. Therefore, an antigenic peptide / immunogenic peptide contains at least one epitope or antigen selected from or derived from a tumor antigen as defined herein.

[0022] Cationic, cationizable: The term “cationic” means that the corresponding structure, compound, group, or atom carries a positive charge, whether permanent or non-permanent, for example, in response to certain conditions such as pH. The terms “cationic,” “cationizable,” and “permanently cationic” as used herein must be understood as defined in WO2023 / 031394 [page 12, line 32 to page 13, line 16].

[0023] Cationic compounds, polycationic compounds: When the corresponding structure, compound, group, or atom carries multiple positive charges, it may be called a polycation. The terminology used herein must be understood as defined in WO2021 / 156267 [page 88, line 12 to page 89, line 22].

[0024] Encoded sequence, encoded region, CDS: As used herein, the term "coding sequence" or the corresponding abbreviation "cds" refers to a sequence of three nucleotides that can be translated into a peptide or protein. In the context of this invention, a cds can be a DNA or RNA sequence consisting of several nucleotides divisible by 3, typically beginning with a start codon and preferably ending with a stop codon. In the context of this invention, a cds encodes at least one tumor antigen as defined herein.

[0025] Derived from: In the context of nucleic acids, the term "derived from" means, for a nucleic acid derived from (another) nucleic acid, that the nucleic acid derived from (another) nucleic acid shares at least 70%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with or is identical to the nucleic acid from which it is derived. In the context of amino acid sequences, the term "derived from" means that the amino acid sequence derived from (another) amino acid sequence shares at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with or is identical to the amino acid sequence from which it is derived.

[0026] Epitope: As used herein, the term "epitope" refers to T-cell epitopes and B-cell epitopes. A T-cell epitope is a portion of an antigenic peptide or protein and may comprise a fragment preferably of about 6 to about 20 or more amino acids in length, such as fragments processed and presented by MHC class I molecules, preferably of about 8 to about 10 amino acids in length, for example, 8, 9, or 10 (or even 11 or 12 amino acids), or fragments processed and presented by MHC class II molecules, preferably of about 13 to about 20 or more amino acids in length. These fragments are typically recognized by T cells as a complex of the peptide fragment and an MHC molecule, i.e., these fragments are not typically recognized in their native form. A B-cell epitope is typically a fragment located on the outer surface of a (native) protein or peptide antigen, preferably having 5 to 15 amino acids, more preferably 5 to 12 amino acids, and even more preferably 6 to 9 amino acids, which can be recognized by antibodies, i.e., in their native form. Such epitopes of proteins or peptides may also be selected from any of the variants of such proteins or peptides mentioned herein. In this context, an epitope can be a conformational epitope or a discontinuous epitope, consisting of segments of a protein or peptide as defined herein that are discontinuous in the amino acid sequence of a protein or peptide as defined herein, but aggregated in a three-dimensional structure; or an epitope can be a continuous epitope or a linear epitope, consisting of a single polypeptide chain.

[0027] Excerpt: The term "fragment" as used herein in the context of nucleic acid sequences (e.g., RNA or DNA) or amino acid sequences can generally refer to a shorter portion of a reference sequence, such as a nucleic acid sequence or amino acid sequence. Therefore, a fragment typically consists of the same sequence as a corresponding stretch within the reference sequence. In the context of this invention, a preferred fragment of a sequence consists of consecutive small segments of an entity, such as nucleotides or amino acids corresponding to consecutive small segments of an entity in the molecule from which the fragment is derived, representing at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the total reference molecule from which the fragment is derived.

[0028] (Sequence) Identity: In the context of nucleic acid or amino acid sequences, the term "identity" refers to the percentage of identical parts between two sequences. To determine the percentage of identical parts between two sequences, the sequences can be aligned (and, if necessary, by introducing gaps) and then compared. In the context of this invention, nucleotides substituted with modified nucleotides (e.g., U replaced by N1-methylpseuuridine (m1ψ)) should not be included in the percentage identity calculation. The percentage of identical parts between two sequences can be determined, for example, using an algorithm, such as one integrated into the BLAST procedure.

[0029] Immunogens, immunogenic: As used herein, the term "immunogen" or "immunogenic" refers to a compound capable of stimulating / inducing an immune response. Preferably, an immunogen is a peptide, polypeptide, or protein.

[0030] RNA in vitro transcription: The term “RNA in vitro transcription” or “in vitro transcription” refers to the process of synthesizing RNA in vitro in a cell-free system. In RNA in vitro transcription (IVT), RNA is obtained by DNA-dependent in vitro transcription of a suitable DNA template in the presence of a DNA-dependent RNA polymerase (e.g., T7, T3, SP6, or Syn5), ribonucleoside triphosphates (NTPs, and optionally modified NTPs); optionally, cap analogs, MgCl2, and buffer (containing TRIS or HEPES, optionally DTT, and / or spermidine).

[0031] (Sequence) variants: Those skilled in the art will recognize and understand that the term "variant" as used herein in the context of nucleic acid sequences is, for example, intended to refer to a variant of a nucleic acid sequence derived from another nucleic acid sequence. For example, a variant of a nucleic acid sequence may exhibit one or more nucleotide deletions, insertions, additions, and / or substitutions compared to the nucleic acid sequence from which it is derived. A variant of a nucleic acid sequence may have at least 50%, 60%, 70%, 80%, 90%, or 95% identity with the nucleic acid sequence from which it is derived. A variant can be a functional variant, meaning that the variant retains at least 50%, 60%, 70%, 80%, 90%, or 95% or more of the function of the sequence from which it is derived. A "variant" of a nucleic acid sequence may have at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% nucleotide identity in a small fragment of at least 30, 50, 75, or 100 nucleotides of the nucleic acid sequence.

[0032] The term "variant" as used herein in the context of proteins or peptides is intended, for example, to refer to a protein or peptide variant having an amino acid sequence that differs from the original sequence in one or more mutations / substitutions, such as one or more substitutions, insertions, and / or deletions of amino acids. Preferably, these fragments and / or variants have the same or comparable specific properties. Insertions and substitutions are possible, particularly in those sequence positions that do not cause changes in the three-dimensional structure or affect the binding region. Changes in the three-dimensional structure by insertion or deletion can be readily determined, for example, using CD spectroscopy (circular dichroism spectroscopy). A variant of a protein can be a functional variant of that protein, meaning that the variant performs at least 40%, 50%, 60%, 70%, 80%, or 90% of the function of a substantially identical protein or a protein from which it is derived. A "variant" of a protein or peptide may have at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% amino acid identity over a small fragment of at least 30, 50, 75, or 100 amino acids of that protein or peptide.

[0033] NSCLC / Non-small cell lung cancer / Lung cancer: Lung cancer (lung carcinoma) is a malignant tumor originating from lung cells. At diagnosis, lung cancer is classified according to the cell type from which the tumor originates; tumors originating from different cells differ in progression and treatment response. Approximately 85% of lung cancers are non-small cell lung cancers, such as adenocarcinoma, squamous cell carcinoma, and large cell carcinoma. NSCLC can be divided into different subgroups, including lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) / squamous non-small cell lung cancer (sqNSCLC).

[0034] Head and neck cancer: It is a type of cancer that commonly occurs in the lips and oral cavity, larynx, salivary glands, nose, sinuses, or facial tissues. The most common type of head and neck cancer occurs in the lips, oral cavity, and larynx. Approximately 90% are pathologically classified as squamous cell carcinoma (HNSCC).

[0035] Squamous cell carcinoma: Squamous cell carcinoma is a group of cancers that typically includes several different types of cancer that originate in squamous cells. These cells form on the surface of the skin, the lining of hollow organs in the body, and the lining of the respiratory and digestive tracts. Squamous cell carcinomas include, for example, squamous cell carcinoma of the head and neck (such as cancers of the mouth, nasal cavity, nasopharynx, throat, and related structures), squamous cell carcinoma of the skin, primary squamous cell carcinoma of the thyroid gland, squamous cell carcinoma of the esophagus, squamous cell carcinoma of the lung, and squamous cell carcinoma of the penis, vagina, ovary, bladder, and eye.

[0036] Melanoma:Melanoma is a type of skin cancer that develops from pigment-producing cells called melanocytes and is the most dangerous type of skin cancer. The outcome of melanoma depends on the stage at which it is diagnosed. Types of melanoma include, for example, cutaneous melanoma (SKCM) or uveal melanoma. Typically, SKCM can be divided into different subgroups, such as the BRAF subgroup and the NRAS subgroup. Detailed Implementation

[0037] Where a reference is made to a “SEQ ID NO” in another patent application or patent, the sequence described therein, such as an amino acid sequence or a nucleic acid sequence, is expressly incorporated herein by reference. For the “SEQ ID NO” provided herein, the information provided under “feature key” in the sequence listing according to WIPO ST.26 standards, namely “source” (for nucleic acids or proteins), “misc_feature” (for nucleic acids), or “REGION” (for proteins), is also expressly included herein. Where a “SEQ ID NO” is mentioned in the context of an RNA sequence, even when a DNA sequence is provided, a person skilled in the art can deduce the RNA sequence from the mentioned SEQ ID NO. Where a “SEQ ID NO” is mentioned in the context of a DNA sequence, even when an RNA sequence is provided, a person skilled in the art can deduce the corresponding DNA sequence from the mentioned SEQ ID NO.

[0038] 1: Nucleic acid encoding at least one tumor antigen:

[0039] In a first aspect, the present invention provides a nucleic acid encoding at least one tumor antigen.

[0040] It is worth noting that the specific features and embodiments described in the context of the first aspect of the invention (i.e., the nucleic acids of the present invention) are equally applicable to any other aspect of the invention, including the pharmaceutical composition of the second aspect, the tumor antigen or composition of the tumor antigen of the third aspect, the antibody, T cell or TCR of the fourth aspect, the combination of the fifth aspect, the kit or kit kit of the sixth aspect, or any aspect relating to medical use or treatment methods.

[0041] In a preferred embodiment, the nucleic acid encoding the at least one tumor antigen is an artificial nucleic acid.

[0042] As used herein, the term "artificial nucleic acid" refers to nucleic acids that are not naturally occurring. In other words, artificial nucleic acids can be understood as non-natural nucleic acid molecules. Such nucleic acid molecules may be non-natural due to their individual sequences (e.g., codon-modified coding sequences, UTRs) and / or other modifications (e.g., structural modifications of nucleotides). Typically, artificial nucleic acids can be genetically engineered and / or generated to correspond to desired artificial nucleotide sequences. In this context, an artificial nucleic acid is a sequence that may not be naturally occurring, i.e., a sequence that differs from a wild-type or naturally occurring sequence by at least one nucleotide. The term "artificial nucleic acid" is not limited to meaning "a single molecule" but is understood to refer to a collection of nucleic acid molecules that are substantially identical. As used herein, the term "artificial nucleic acid" can refer to artificial DNA or artificial RNA. Preferably, the artificial nucleic acid is selected from artificial RNA.

[0043] In a preferred embodiment, the artificial nucleic acid contains at least one coding sequence that encodes at least one tumor antigen.

[0044] The term "tumor antigen" used in this article refers to tumor-associated antigens (TAAs) expressed by normal and tumor tissues, and tumor-specific antigens, including shared antigens, neoantigens, and unique antigens. Other tumor antigens can be broadly classified as carcinoembryonic antigens (typically expressed in fetal tissues and cancerous somatic cells), oncoviral antigens (encoded by tumorigenic transforming viruses), overexpressed / accumulated antigens (expressed in normal and tumor tissues, but with highly elevated levels in tumors), cancer-testis antigens (expressed only by cancer cells and adult reproductive tissues such as the testes and placenta), lineage-restricted antigens (primarily expressed by a single cancer histotype), mutant antigens (expressed only in cancer due to gene mutations or transcriptional alterations), post-translational altered antigens (tumor-related alterations such as glycosylation), or unique antigens (highly polymorphic genes in which tumor cells express a specific "clonal type," i.e., B-cell or T-cell lymphomas / leukemias caused by clonal abnormalities). It is important to emphasize that these categories are not mutually exclusive, and a tumor antigen may belong to more than one category.

[0045] In a preferred embodiment, the tumor antigen (provided by artificial nucleic acid) is generated in the cytoplasm after the artificial nucleic acid is applied to cells, tissues, or objects. The generated tumor antigen may be secreted or further processed.

[0046] Therefore, the application of artificial nucleic acids (e.g., RNA) to cells, tissues, or objects results in the translation of at least one coding sequence into at least one tumor antigen. Whenever "generated tumor antigen" is mentioned, the term refers to a protein product generated from the artificial nucleic acids of the present invention by translating the coding sequence of the nucleic acid into a protein. Therefore, the functional and structural features and embodiments described herein in relation to "tumor antigen" or "(generated) tumor antigen" should be understood as referring to the tumor antigen amino acid sequence generated / translated in the cytoplasm after the application of the artificial nucleic acid to cells, tissues, or objects.

[0047] In an embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA), or immunogenic fragments or variants of such peptides or proteins.

[0048] In an embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from or derived from peptides or proteins of tumor neoantigens, or immunogenic fragments or variants of such peptides or proteins.

[0049] The terms “immunogenic fragment” or “immunogenic variant” must be understood as any fragment / variant of the corresponding peptide or protein that is capable of evoking an immune response in a subject, preferably a human subject.

[0050] In the implementation scheme, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting:

[0051] a) At least one amino acid sequence encoded by a long non-coding RNA (lncRNA), or an immunogenic fragment or variant thereof; and / or

[0052] b) The amino acid sequence of at least one tumor neoantigen, or its immunogenic fragment or variant.

[0053] In the implementation scheme, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting:

[0054] a) At least one antigenic peptide, said antigenic peptide being selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA), or immunogenic fragments or variants thereof; and / or

[0055] b) At least one antigenic peptide, said antigenic peptide being selected from or derived from peptides or proteins of tumor neoantigens, or immunogenic fragments or variants thereof.

[0056] In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA), or immunogenic fragments or variants thereof.

[0057] Tumor antigens encoded by long non-coding RNAs (lncRNAs)

[0058] In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA), or immunogenic fragments or variants thereof.

[0059] The term "long non-coding RNA" and its abbreviation "lncRNA" refer to RNAs that are typically longer than 200 bases (up to, for example, 100,000 bases). Previously, lncRNAs were thought to lack protein-coding function. Recent studies of long non-coding RNAs have shown that these molecules are involved in various aspects of cellular regulation. Compared to mRNA, long non-coding RNAs are characterized by lower expression abundance but higher tissue and cell expression specificity. Recently, several lncRNAs have been found to encode peptides with biologically significant functions, such as micropeptides or putative small open reading frames (also referred to herein as "mini ORFs" or "small ORFs," smORFs). Therefore, due to the nomenclature commonly used in this field, the term "long non-coding RNA" or its abbreviation "lncRNA" is used throughout this specification. It is worth noting that, in the context of this invention, lncRNA species encode peptides or proteins that can serve as suitable tumor antigens.

[0060] Surprisingly, the inventors have identified peptides generated by certain types of lncRNAs. Even more surprisingly, these lncRNA-encoded peptides exhibit specificity for certain tumors (e.g., melanoma, NSCLC, HNSCC) and can elicit antigen-specific immune responses. For example, these peptides can be recognized as foreign substances by immune cells, presented, and trigger antigen-specific immune responses (e.g., T-cell responses). Therefore, peptides selected from or derived from lncRNA-based smORFs are suitable for cancer immunotherapy.

[0061] In a preferred embodiment, at least one antigenic peptide is selected from amino acid sequences, such as peptides or proteins encoded by long non-coding RNA (lncRNA), or fragments or variants thereof, wherein the lncRNA contains at least one smORF (encoding the peptide or protein), and wherein the lncRNA is characterized by increased expression in cancer cells (e.g., melanoma, NSCLC, HNSCC) compared to non-cancer cells. Furthermore, the lncRNA contains at least one smORF.

[0062] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by long non-coding RNA, or immunogenic fragments or variants thereof, wherein the lncRNA is selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, or any of these fragments or variants, preferably any of these immunogenic fragments or variants.

[0063] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence encoded by a long non-coding RNA, or a fragment or variant thereof, wherein the lncRNA is selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, or any fragment or variant thereof, preferably any of these immunogenic fragments or variants.

[0064] The terms “amino acid sequence encoded by…”, “peptide or protein encoded by…”, or “antigen encoded by…” define, in the context of a specific lncRNA species (e.g., encoded by ZC3H8-6:1, WDR72-2:4, KCNMB2-AS1:4, or NTF3-5:5), that the origin or source of the corresponding amino acid sequence, peptide, protein, or antigen is a long non-coding RNA. Thus, in other words, the corresponding amino acid sequence, peptide, protein, or antigen originates from the corresponding lncRNA species. In the context of this invention, the corresponding amino acid sequence, peptide, protein, or antigen encoded by a specific lncRNA species (e.g., ZC3H8-6:1, WDR72-2:4, KCNMB2-AS1:4, or NTF3-5:5), or in other words, derived from a specific lncRNA species (e.g., ZC3H8-6:1, WDR72-2:4, KCNMB2-AS1:4, or NTF3-5:5), is provided by the nucleic acid (e.g., mRNA) of this invention.

[0065] In the context of this invention, preferred are, for example, amino acid sequences encoding peptides or proteins, or immunogenic fragments or variants thereof, of lnc-NTF3-5:5, lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-ZC3H8-6:1, or lnc-CLEC2D-9:1, or nucleic acid sequences encoding, for example, amino acid sequences of peptides or proteins, or fragments or variants thereof, of lnc-NTF3-5:5, lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-ZC3H8-6:1, or lnc-CLEC2D-9:1.

[0066] In the context of this invention, amino acid sequences, such as peptides or proteins encoded by lnc-NTF3-5:5, or immunogenic fragments or variants thereof, are particularly preferred, or nucleic acid sequences encoding, for example, peptides or proteins encoded by lnc-NTF3-5:5, or fragments or variants thereof.

[0067] Importantly, as shown by RNA sequencing (see Examples section), lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, and LINC00893:25 are primarily expressed in cancer cells, and peptides encoded by these lncRNAs have been identified by mass spectrometry (see Examples section).

[0068] lnc-WDR72-2:4 is a long non-coding RNA transcribed in humans from the following genomic location: chr15,53513740,53540852,-. The naturally occurring (human) lncRNA sequence of lnc-WDR72-2:4 is shown in SEQ ID NO: 512. The RNA sequence of lnc-WDR72-2:4 contains at least two open reading frames (smORF lncWDR722416421696 and smORF lncWDR722415641696) that encode at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from lnc-WDR72-2:4, or an immunogenic fragment or variant thereof, can serve as a suitable tumor antigen in the context of this invention.

[0069] lnc-TRPC5-3:1 is a long non-coding RNA transcribed in humans from the following genomic location: chrX,111619483,111621517,-. The naturally occurring (human) lncRNA sequence of lnc-TRPC5-3:1 is shown in SEQ ID NO:516. The RNA sequence of lnc-TRPC5-3:1 contains at least one open reading frame (smORFlncTRPC53116031801) encoding at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from lnc-TRPC5-3:1, or its immunogenic fragments or variants, can serve as a suitable tumor antigen in the context of this invention.

[0070] lnc-CLEC2D-9:1 is a long non-coding RNA transcribed in humans from the following genomic location: chr12,9570760,9576267,+. The naturally occurring (human) lncRNA sequence of lnc-CLEC2D-9:1 is shown in SEQ ID NO: 517. The RNA sequence of lnc-CLEC2D-9:1 contains at least one open reading frame (smORF lncCLEC2D91412616) encoding at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from lnc-CLEC2D-9:1, or its immunogenic fragments or variants, can serve as a suitable tumor antigen in the context of this invention.

[0071] KCNMB2-AS1:4 is a long non-coding RNA transcribed in humans from the following genomic locations: chr3,178525486,178860397,-. The naturally occurring (human) lncRNA sequence of KCNMB2-AS1:4 is shown in SEQ ID NO:513. The RNA sequence of KCNMB2-AS1:4 contains at least one open reading frame (smORF KCNMB2AS14633753) encoding at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from KCNMB2-AS1:4, or its immunogenic fragments or variants, can serve as a suitable tumor antigen in the context of this invention.

[0072] lnc-ZC3H8-6:1 is a long non-coding RNA transcribed in humans from the following genomic locations: chr2,111769354,111776910,-. The naturally occurring (human) lncRNA sequence of lnc-ZC3H8-6:1 is shown in SEQ ID NO:514. The RNA sequence of lnc-ZC3H8-6:1 contains at least one open reading frame (smORF lncZC3H86163144) encoding at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from lnc-ZC3H8-6:1, or its immunogenic fragments or variants, can serve as a suitable tumor antigen in the context of this invention.

[0073] lnc-NTF3-5:5 is a long non-coding RNA transcribed in humans from the following genomic location: chr12,5367810,5379568,+. The naturally occurring (human) lncRNA sequence of lnc-NTF3-5:5 is shown in SEQ ID NO: 515. The RNA sequence of lnc-NTF3-5:5 contains at least one open reading frame (smORF lncNTF35511951417) encoding at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from lnc-NTF3-5:5, or its immunogenic fragments or variants, can serve as a suitable tumor antigen in the context of this invention.

[0074] LINC00893:25 (encoding smORF LINC00893) is a long non-coding RNA transcribed in humans from the following genomic locations: chrX,149483112,149532914,-. The naturally occurring (human) lncRNA sequence of LINC00893:25 is shown in SEQ ID NO: 518. The RNA sequence of LINC00893:25 contains at least one open reading frame (smORF LINC00893) encoding at least one amino acid sequence, such as at least one peptide that can serve as a tumor antigen in the context of this invention. Suitably, any peptide or protein derived from LINC00893:25, or its immunogenic fragments or variants, can serve as a suitable tumor antigen in the context of this invention.

[0075] Preferably, at least one antigenic peptide encoded by a long non-coding RNA, or any immunogenic fragment or variant thereof, contains at least one T-cell epitope, preferably at least one CD8+ T-cell epitope.

[0076] Preferably, at least one antigenic peptide, or any immunogenic fragment or variant thereof, encoded by long non-coding RNA lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, contains at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0077] In a preferred embodiment, at least one antigenic peptide encoded by a long non-coding RNA, or any immunogenic fragment or variant thereof, induces epitope-specific T cells in the subject, preferably epitope-specific CD8+ T cells in the subject (particularly human subjects).

[0078] In a particularly preferred embodiment, at least one antigenic peptide encoded by long non-coding RNA lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, or any of these immunogenic fragments or variants, induces epitope-specific T cells in the subject, preferably epitope-specific CD8+ T cells in the subject (particularly human subjects).

[0079] The term "T-cell epitope" refers to a portion or fragment of a protein or antigen that is recognized by T cells when presented in the context of major histocompatibility complex molecules. This includes MHC class I and MHC class II molecules and involves gene complexes present in all vertebrates. MHC proteins or molecules are important for signaling in the immune response between lymphocytes and antigen-presenting cells or diseased cells, where MHC proteins or molecules bind peptide epitopes and present them for recognition by T-cell receptors on T cells. MHC-encoded proteins are expressed on the cell surface and present T cells with both self-antigens (peptide fragments from the cell itself) and non-self-antigens (e.g., fragments of invading microorganisms). For class I MHC / peptide complexes, the binding peptide (MHC class I ligand) is typically about 8 to about 10 amino acids long, although longer or shorter peptides may be effective. For class II MHC / peptide complexes, the binding peptide (MHC class II ligand) is typically about 10 to about 25 amino acids long, particularly about 13 to about 18 amino acids, although longer and shorter peptides may also be effective.

[0080] Preferably, at least one antigenic peptide encoded by lncRNA, or an immunogenic fragment or variant thereof, particularly lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, is an antigen or epitope exhibiting HLA binding affinity or MHC binding affinity.

[0081] As used herein, the term "HLA" or "human leukocyte antigen" includes variants, allotypes, and species homologs of HLA, as well as analogs having at least one common epitope with HLA. As used herein, "human leukocyte antigen (HLA)" should preferably be understood as a genetic complex encoding proteins of the major histocompatibility complex (MHC) in humans. These cell surface proteins are responsible for the regulation of the human immune system. HLA genes are highly polymorphic, meaning they have many different alleles, which allows them to fine-tune an individual's adaptive immune system. In the context of this description, the term "HLA binding affinity" or "MHC binding affinity" should be understood as the binding affinity between a specific antigen and a specific MHC allele. In the context of this description, the term "HLA type" should be understood as the complete set of HLA gene alleles.

[0082] Preferably, at least one antigenic peptide encoded by lncRNA, as specified herein, is capable of binding to an antibody or a T-cell receptor. Preferably, the antigenic peptide contains at least one MHC class I or MHC class II ligand.

[0083] In the implementation scheme, at least one antigenic peptide encoded by lncRNA as defined herein is a shared tumor-associated antigen, particularly a shared tumor-associated antigen in NSCLC and / or melanoma.

[0084] In a preferred embodiment, at least one antigenic peptide (provided by an artificial nucleic acid) is selected from peptides or proteins encoded by long non-coding RNA (lncRNA) as defined herein, and comprises or constitutes at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these.

[0085] Therefore, at least one antigenic peptide preferably comprises at least one amino acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, or an immunogenic fragment or variant thereof.

[0086] In this context, the immunogenic fragments of SEQ ID NO: 124 to SEQ ID NO: 254, SEQ ID NO: 546 to SEQ ID NO: 650, and SEQ ID NO: 756 to SEQ ID NO: 763 have shorter amino acid sequences than the corresponding reference sequences. Therefore, the immunogenic fragments of SEQ ID NO: 124 to SEQ ID NO: 254, SEQ ID NO: 546 to SEQ ID NO: 650, and SEQ ID NO: 756 to SEQ ID NO: 763 are truncated at the N-terminus by, for example, 1, 5, 10, or more than 10 amino acids and / or truncated at the C-terminus by, for example, 1, 5, 10, or more than 10 amino acids. It is noteworthy that the immunogenic fragments still contain at least one epitope capable of evoking an immune response, preferably at least one T-cell epitope. In an embodiment, at least one epitope has the size / length of an MHC class I or MHC class II epitope. In a preferred embodiment, at least one epitope has the length of an MHC class I epitope. In some embodiments, at least one epitope has the length of an MHC class II epitope but contains at least one MHC class I epitope and can be processed during antigen processing during MHC:peptide complex loading. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids.

[0087] lnc-WDR72-2:4

[0088] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by lnc-WDR72-2:4 (smORF lncWDR722416421696 or smORF lncWDR722415641696), or an immunogenic fragment or variant thereof.

[0089] The full-length polypeptide encoded by lncRNA lnc-WDR72-2:4 (SEQ ID NO: 124 and SEQ ID NO: 144) contains several predicted epitopes according to SEQ ID NO: 125 to SEQ ID NO: 143, SEQ ID NO: 145 to SEQ ID NO: 146, and SEQ ID NO: 546 to SEQ ID NO: 571. Therefore, full-length polypeptides containing epitopes as defined herein (smORF lncWDR722416421696 or lncWDR722415641696 according to SEQ ID NO: 124 or SEQ ID NO: 144) or any fragment or variant thereof are particularly useful in the context of this invention.

[0090] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-WDR72-2:4, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, and SEQ ID NO: 757.

[0091] Therefore, at least one antigenic peptide preferably comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, or SEQ ID NO: 757.

[0092] In this context, the fragments may be N-terminal truncated from SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, or SEQ ID NO: 757, or C-terminal truncated from SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, or SEQ ID NO: 757. In an embodiment, the immunogenic fragments of SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, and SEQ ID NO: 757 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, and SEQ ID NO: 757 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. In this context, the typical size of the immunogenic fragment is 5 to 20 amino acids, or 8 to 15 amino acids.

[0093] In a particularly preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-WDR72-2:4, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 756.

[0094] lnc-TRPC5-3:1

[0095] In an embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by lnc-TRPC5-3:1 (smORFlncTRPC53116031801), or an immunogenic fragment or variant thereof.

[0096] The full-length polypeptide encoded by lncRNA TRPC5-3:1 (SEQ ID NO: 227) contains several predicted epitopes according to SEQ ID NO: 228 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642. Therefore, full-length polypeptides containing epitopes as defined herein (smORF lncTRPC53116031801 according to SEQ ID NO: 227) or any fragments or variants thereof are particularly useful in the context of this invention.

[0097] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-TRPC5-3:1, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, or SEQ ID NO: 761.

[0098] Therefore, at least one antigenic peptide preferably comprises or consists of at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, or SEQ ID NO: 761.

[0099] In this context, the fragments may be N-terminal truncated from SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, or SEQ ID NO: 761, or C-terminal truncated from SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, or SEQ ID NO: 761. In an embodiment, the immunogenic fragments of SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, or SEQ ID NO: 761 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, and SEQ ID NO: 761 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. In this context, the typical size of the immunogenic fragment is 5 to 20 amino acids, or 8 to 15 amino acids.

[0100] In a preferred embodiment, at least one antigenic peptide is selected from a peptide or protein encoded by lnc-TRPC5-3:1, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to SEQ ID NO: 761 or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity.

[0101] lnc-CLEC2D-9:1

[0102] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by lnc-CLEC2D-9:1 (smORF lncCLEC2D91412616), or an immunogenic fragment or variant thereof.

[0103] The full-length polypeptide encoded by lncRNA lnc-CLEC2D-9:1 (SEQ ID NO: 238) contains several predicted epitopes according to SEQ ID NO: 239 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650. Therefore, full-length polypeptides containing epitopes as defined herein (smORF lncCLEC2D91412616 according to SEQ ID NO: 238) or any fragments or variants thereof are particularly useful in the context of this invention.

[0104] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-CLEC2D-9:1 and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, or SEQ ID NO: 762.

[0105] Therefore, at least one antigenic peptide preferably comprises or consists of at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, or SEQ ID NO: 762.

[0106] In this context, the immunogenic fragment may be an N-terminal truncated portion of SEQ ID NO: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, or SEQ ID NO: 762, or a C-terminal truncated portion of SEQ ID NO: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, or SEQ ID NO: 762. In an embodiment, the immunogenic fragments of SEQ ID NO: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, or SEQ ID NO: 762 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID No: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, and SEQ ID NO: 762 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. In this context, the typical size of the immunogenic fragment is 5 to 20 amino acids, or 8 to 15 amino acids.

[0107] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-CLEC2D-9:1, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to SEQ ID NO: 762 or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity.

[0108] KCNMB2-AS1:4

[0109] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by KCNMB2-AS1:4 (smORF KCNMB2AS14633753), or an immunogenic fragment or variant thereof.

[0110] The full-length polypeptide encoded by lncRNA KCNMB2-AS1:4 (SEQ ID NO: 147) contains several predicted epitopes according to SEQ ID NO: 148 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589. Therefore, full-length polypeptides containing epitopes as defined herein (smORF KCNMB2AS14633753 according to SEQ ID NO: 147) or any fragments or variants thereof are particularly useful in the context of this invention.

[0111] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by KCNMB2-AS1:4, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, or SEQ ID NO: 758.

[0112] Therefore, at least one antigenic peptide preferably comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID No: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, or SEQ ID NO: 758.

[0113] In this context, the fragments may be N-terminal truncated from SEQ ID NO: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, or SEQ ID NO: 758, or C-terminal truncated from SEQ ID NO: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, or SEQ ID NO: 758. In embodiments, the immunogenic fragments of SEQ ID NO: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, and SEQ ID NO: 758 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NOs: 147 to 161, 572 to 589, and 758 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. In this context, the typical size of the immunogenic fragment is 5 to 20 amino acids, or 8 to 15 amino acids.

[0114] In a particularly preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by KCNMB2-AS1:4 and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 758.

[0115] lnc-ZC3H8-6:1

[0116] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by lnc-ZC3H8-6:1 (smORF lncZC3H86163144), or an immunogenic fragment or variant thereof.

[0117] The full-length polypeptide encoded by lncRNA lnc-ZC3H8-6:1 (SEQ ID NO: 162) contains several predicted epitopes according to SEQ ID NO: 163 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603. Therefore, full-length polypeptides containing epitopes as defined herein (smORF lncZC3H86163144 according to SEQ ID NO: 162) or any fragments or variants thereof are particularly useful in the context of this invention.

[0118] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-ZC3H8-6:1, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 162 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603, or SEQ ID NO: 759.

[0119] Therefore, at least one antigenic peptide preferably comprises or consists of at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 162 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603, or SEQ ID NO: 759.

[0120] In this context, the fragments may be N-terminal truncated from SEQ ID NO: 162 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603, or SEQ ID NO: 759, or C-terminal truncated from SEQ ID NO: 162 to SEQ ID NO: 168. In embodiments, the immunogenic fragments of SEQ ID NO: 162 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603, and SEQ ID NO: 759 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 162 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603, and SEQ ID NO: 759 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. In this context, the typical size of the immunogenic fragment is 5 to 20 amino acids, or 8 to 15 amino acids.

[0121] In a particularly preferred embodiment, at least one antigenic peptide is selected from a peptide or protein encoded by lnc-ZC3H8-6:1, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 759.

[0122] lnc-NTF3-5:5

[0123] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by lnc-NTF3-5:5 (smORF lncNTF35511951417), or an immunogenic fragment or variant thereof.

[0124] The full-length polypeptide encoded by lncRNA lnc-NTF3-5:5 (SEQ ID NO: 169) contains several predicted epitopes according to SEQ ID NO: 170 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629. Therefore, full-length polypeptides containing epitopes as defined herein (smORF lncNTF35511951417 according to SEQ ID NO: 169) or any fragments or variants thereof are particularly useful in the context of this invention.

[0125] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-NTF3-5:5, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 169 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629, or SEQ ID NO: 760.

[0126] Therefore, at least one antigenic peptide preferably comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 169 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629, or SEQ ID NO: 760.

[0127] In this context, the fragments may be N-terminal truncated from SEQ ID NO: 169 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629, or SEQ ID NO: 760, or C-terminal truncated from SEQ ID NO: 169 to SEQ ID NO: 226. In embodiments, the immunogenic fragments of SEQ ID NO: 169 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629, and SEQ ID NO: 760 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 169 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629, and SEQ ID NO: 760 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. In this context, the typical size of the immunogenic fragment is 5 to 20 amino acids, or 8 to 15 amino acids.

[0128] In a particularly preferred embodiment, at least one antigenic peptide is selected from a peptide or protein encoded by lnc-NTF3-5:5, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 760.

[0129] LINC00893:25

[0130] In an embodiment, at least one antigenic peptide comprises an amino acid sequence, such as a peptide or protein encoded by LINC00893:25 (LINC00893), or an immunogenic fragment or variant thereof.

[0131] Full-length polypeptides containing epitopes as defined herein (smORF LINC00893 according to SEQ ID NO: 244) or any fragments or variants thereof are particularly useful in the context of this invention. Thus, the full-length polypeptide encoded by lncRNA LINC00893 (SEQ ID NO: 244) contains several predicted epitopes according to SEQ ID NO: 245 to SEQ ID NO: 254.

[0132] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by LINC00893:25, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 244 to SEQ ID NO: 254 and SEQ ID NO: 763.

[0133] Therefore, at least one antigenic peptide preferably comprises or consists of at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 244 to SEQ ID NO: 254 and SEQ ID NO: 763.

[0134] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 244 to 254, SEQ ID NO: 763, or a C-terminal truncation of SEQ ID NO: 244 to SEQ ID NO: 254, SEQ ID NO: 763. In embodiments, the immunogenic fragments of SEQ ID NO: 244 to SEQ ID NO: 254, SEQ ID NO: 763 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 244 to SEQ ID NO: 254, SEQ ID NO: 763 still contain at least one epitope, preferably a T-cell epitope, capable of evoking an immune response. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0135] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by LINC00893:25, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, which is identical to SEQ ID NO: 763 or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence.

[0136] Preferred nucleic acid sequences encoding antigens encoded by lncRNA

[0137] The following describes the nucleic acid coding sequence that encodes the antigenic peptide specified herein (encoded by the corresponding lncRNA), which may be contained in at least one cds of an artificial nucleic acid.

[0138] In a preferred embodiment, at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins encoded by lncRNAs, wherein the nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants. In this context, preferably, the fragment is at least 24 nucleotides in length.

[0139] Suitably, the nucleic acid sequence encoding at least one antigenic peptide selected from lncRNA-encoded peptides or proteins is a codon-modified CDS, preferably wherein the codon-modified CDS is selected from C-maximized CDS (as further defined herein), CAI-maximized CDS (as further defined herein), human codon selection adapted CDS (as further defined herein), G / C content modified CDS (as further defined herein), and G / C optimized CDS (as further defined herein), or any combination thereof. In the preferred embodiment within this context, the coding sequence modified with at least one codon is a G / C optimized coding sequence.

[0140] Therefore, in a particularly preferred embodiment, at least one coding sequence comprises a GC-optimized nucleic acid sequence encoding at least one antigenic peptide selected from lncRNA-encoded peptides or proteins, wherein the nucleic acid sequence is identical to or has at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments. In this context, preferably, the fragment is at least 24 nucleotides in length.

[0141] In a particularly preferred embodiment, at least one artificial nucleic acid comprises

[0142] - At least one CDS comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-NTF3-5:5, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, preferably SEQ ID NO: 768, or a fragment or variant thereof; or

[0143] - At least one CDS comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-WDR72-2:4, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 318 to SEQ ID NO: 340, SEQ ID NO: 651 to SEQ ID NO: 676, SEQ ID NO: 764, SEQ ID NO: 765, or any fragment or variant thereof, preferably SEQ ID NO: 765; or

[0144] - At least one CDS comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-TRPC5-3:1, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, preferably SEQ ID NO: 769, or a fragment or variant thereof; or

[0145] - At least one CDS containing a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen containing at least one antigenic peptide selected from a peptide or protein encoded by lnc-CLEC2D-9:1, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, preferably SEQ ID NO: 770, or a fragment or variant thereof; or

[0146] - At least one CDS containing a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by KCNMB2-AS1:4, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, preferably SEQ ID NO: 766, or a fragment or variant thereof; or

[0147] - At least one CDS comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-ZC3H8-6:1, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, preferably SEQ ID NO: 767, or a fragment or variant thereof; or

[0148] - At least one CDS containing a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen containing at least one antigenic peptide selected from the peptide or protein encoded by LINC00893:25, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 438 to SEQ ID NO: 448, SEQ ID NO: 771, or any fragment or variant thereof, preferably SEQ ID NO: 771, or a fragment or variant thereof.

[0149] Preferred amino acid and nucleic acid sequences in this context are provided in Table 1A. Each row corresponds to a suitable antigenic peptide or protein encoded by a long non-coding RNA (lncRNA). Column A provides a brief description of the corresponding lncRNA encoding the tumor antigen. Column B provides the SEQ ID NO of the corresponding full-length amino acid sequence encoded by the lncRNA. Column C provides the SEQ ID NO of the corresponding fragment or epitope. Column D provides the SEQ ID NO of the G / C-optimized nucleic acid sequence encoding the full-length amino acid sequence in column B. Column E provides the SEQ ID NO of the G / C-optimized nucleic acid sequence encoding the corresponding fragment or epitope in column C. Column F shows the preferred cancer indications for the antigen (N = NSCLC; M = melanoma; H = HNSCC; see also aspect 7 (medical use) of cancer indications further provided, particularly for antigens encoded by lncRNAs in rows 1 through 4 and 6).

[0150] Table 1A: Preferred antigens encoded by lncRNAs (amino acid sequence and coding sequence) )

[0151]

[0152] In this context, particularly preferred amino acid and nucleic acid sequences are provided in Table 1B. Each row corresponds to a particularly suitable antigenic peptide or protein encoded by a long non-coding RNA (lncRNA). Column A provides a brief description of the corresponding lncRNA encoding a tumor antigen. Column B provides the SEQ ID NO of the corresponding full-length amino acid sequence encoded by that lncRNA. Column C provides the SEQ ID NO of the preferred fragment or epitope. Column D provides the SEQ ID NO of the G / C-optimized nucleic acid sequence encoding the full-length amino acid sequence in column B. Column E provides the SEQ ID NO of the G / C-optimized nucleic acid sequence encoding the preferred fragment or epitope in column C.

[0153] Table 1B: Particularly Preferred Antigens Encoded by lncRNAs (Amino Acid Sequences and Coding Sequences)

[0154]

[0155] In a particularly preferred embodiment, at least one antigenic peptide comprises or consists of at least one amino acid sequence selected from columns B or C of Table 1A or Table 1B, or any of these immunogenic fragments.

[0156] In a particularly preferred embodiment, the at least one coding sequence of the artificial nucleic acid of the present invention comprises or constitutes at least one nucleic acid sequence, or any fragment or variant thereof, provided in column D or column E of Table 1A or Table 1B.

[0157] Tumor antigens selected from tumor neoantigens

[0158] In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from or derived from peptides or proteins of tumor neoantigens, or immunogenic fragments or variants thereof.

[0159] The term "tumor neoantigen" refers to specific polypeptides produced by gene mutations in tumor cells. These polypeptides are presented to the cell surface via MHC molecules (HLA molecules in mammals), where they are recognized by T lymphocytes to trigger an immune response. Therefore, neoantigens are suitable as biomarkers for immunotherapy and can be further used to develop tumor neoantigen vaccines, as these neoantigens are produced by mutations in tumor cells but are not expressed in normal or healthy cells. Thus, neoantigens may be promising targets for immunotherapy.

[0160] In a preferred embodiment, the neoantigen is a shared neoantigen, preferably a shared neoantigen for patients with NSCLC and / or melanoma cancer.

[0161] Surprisingly, peptides present in tumor cells have been identified. Even more surprisingly, these peptides exhibit specificity in tumor cells and elicit immune responses, preferably antigen-specific immune responses.

[0162] In the implementation scheme, at least one antigenic peptide selected from tumor neoantigens is selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, PIK3CA neoantigen, NFE2L2 neoantigen, CNOT9 neoantigen, or any immunogenic fragment or variant of these.

[0163] For clarity, the terms “NRAS neoantigen,” “TYW1B neoantigen,” “ECPAS neoantigen,” “MAP2K1 neoantigen,” “TOMM22 neoantigen,” “GLB1 neoantigen,” “MAGE-A3 neoantigen,” “ATAD2 neoantigen,” “BRAF neoantigen,” “EGFR neoantigen,” “TP53 neoantigen,” “DIP2B neoantigen,” “TRRAP neoantigen,” “RAC1 neoantigen,” “AP4B1 neoantigen,” “VWA5A neoantigen,” “CLTC neoantigen,” “RPL9 neoantigen,” “RPAP1 neoantigen,” “RPE65 neoantigen,” “PATZ1 neoantigen,” “PIK3CA neoantigen,” “NFE2L2 neoantigen,” and “CNOT9 neoantigen” refer to the corresponding mutant proteins found in cancer cells, but not the corresponding wild-type or somatic proteins.

[0164] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, or any immunogenic fragments or variants of these.

[0165] In a preferred embodiment, at least one antigenic peptide selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, and PATZ1 neoantigen contains at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0166] In a particularly preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of tumor neoantigens, or immunogenic fragments or variants thereof, particularly peptides or proteins of NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, or immunogenic fragments or variants thereof, to induce epitope-specific T cells in the subject, preferably epitope-specific CD8+ T cells in the subject (particularly human subjects).

[0167] In a preferred embodiment, at least one antigenic peptide, or immunogenic fragment or variant thereof, selected from a peptide or protein of a tumor neoantigen, is an antigen or epitope exhibiting HLA binding affinity or MHC binding affinity.

[0168] Preferably, at least one antigenic peptide of the tumor neoantigen specified herein binds to an antibody or a T-cell receptor. In a preferred embodiment, the antigenic peptide comprises an MHC class I or MHC class II ligand / epitope.

[0169] In the embodiments, at least one antigenic peptide (provided by the artificial nucleic acid) is selected from peptides or proteins of tumor neoantigens and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 255 to SEQ ID NO: 317.

[0170] In a preferred embodiment, at least one antigenic peptide (provided by the artificial nucleic acid) is selected from peptides or proteins of tumor neoantigens and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 255 to SEQ ID NO: 286.

[0171] In this context, the immunogenic fragments of SEQ ID NO: 255 to SEQ ID NO: 317 have a shorter amino acid sequence than the corresponding reference sequence. The immunogenic fragments of SEQ ID NO: 255 to SEQ ID NO: 317 are truncated at the N-terminus by, for example, 1, 5, 10, or more than 10 amino acids and / or truncated at the C-terminus by, for example, 1, 5, 10, or more than 10 amino acids. Notably, the immunogenic fragments still contain at least one epitope capable of evoking an immune response, preferably at least one T-cell epitope. In some embodiments, at least one epitope has the size / length of an MHC class I or MHC class II epitope. In a preferred embodiment, at least one epitope has the length of an MHC class I epitope. In some embodiments, at least one epitope has the length of an MHC class II epitope but contains at least one MHC class I epitope and can be processed during antigen processing during MHC:peptide complex loading. Typical immunogenic fragment sizes in this context are 5 to 20 amino acids, or 8 to 15 amino acids.

[0172] NRAS amino acid and nucleic acid sequences

[0173] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of NRAS tumor neoantigens and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 267 to SEQ ID NO: 272. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of NRAS tumor neoantigens, said antigenic peptide comprising or constituting at least one amino acid sequence, or an immunogenic fragment or variant thereof, said amino acid sequence being identical to or having at least 80% identity with any of SEQ ID NO: 267 to SEQ ID NO: 272.

[0174] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 267 to SEQ ID NO: 272 or a C-terminal truncation of SEQ ID NO: 267 or SEQ ID NO: 272. In embodiments, the immunogenic fragments of SEQ ID NO: 267 to SEQ ID NO: 272 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 267 to SEQ ID NO: 272 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0175] TYW1B amino acid and nucleic acid sequence

[0176] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the TYW1B tumor neoantigen, and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 273 or SEQ ID NO: 274. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of the TYW1B tumor neoantigen, wherein said antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 80% identity with any of SEQ ID NO: 273 or SEQ ID NO: 274.

[0177] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 273 or SEQ ID NO: 274, or a C-terminal truncation of SEQ ID NO: 273 or SEQ ID NO: 274. In embodiments, the immunogenic fragments of SEQ ID NO: 273 or SEQ ID NO: 274 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 273 or SEQ ID NO: 274 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0178] ECPAS amino acid and nucleic acid sequences

[0179] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of ECPAS tumor neoantigens and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 275. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, wherein the tumor antigen comprises or constitutes at least one antigenic peptide selected from peptides or proteins of ECPAS tumor neoantigens, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 80% identity with SEQ ID NO: 275.

[0180] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 275 or a C-terminal truncation of SEQ ID NO: 275. In embodiments, the immunogenic fragment of SEQ ID NO: 275 is truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragment of SEQ ID NO: 275 still contains at least one epitope capable of evoking an immune response, preferably a T-cell epitope. A typical immunogenic fragment in this context is 5 to 20 amino acids in size, or 8 to 15 amino acids.

[0181] MAP2K1 amino acid and nucleic acid sequence

[0182] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the MAP2K1 tumor neoantigen and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 276 or SEQ ID NO: 277. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, wherein the tumor antigen comprises or constitutes at least one antigenic peptide selected from peptides or proteins of the MAP2K1 tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 80% identity with any of SEQ ID NO: 276 or SEQ ID NO: 277.

[0183] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 276 or SEQ ID NO: 277, or a C-terminal truncation of SEQ ID NO: 276 or SEQ ID NO: 277. In embodiments, the immunogenic fragments of SEQ ID NO: 276 or SEQ ID NO: 277 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 276 or SEQ ID NO: 277 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0184] TOMM22 amino acid and nucleic acid sequence

[0185] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the TOMM22 tumor neoantigen and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 278 or SEQ ID NO: 279. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of the TOMM22 tumor neoantigen, wherein said antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 80% identity with any of SEQ ID NO: 278 or SEQ ID NO: 279.

[0186] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 278 or SEQ ID NO: 279, or a C-terminal truncation of SEQ ID NO: 278 or SEQ ID NO: 279. In embodiments, the immunogenic fragments of SEQ ID NO: 278 or SEQ ID NO: 279 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 278 or SEQ ID NO: 279 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0187] GLB1 amino acid and nucleic acid sequence

[0188] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the GLB1 tumor neoantigen and comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 280 or SEQ ID NO: 281. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of the GLB1 tumor neoantigen, or immunogenic fragments or variants thereof, wherein said antigenic peptide comprises or constitutes at least one amino acid sequence, said amino acid sequence being identical to or having at least 80% identity with any of SEQ ID NO: 280 or SEQ ID NO: 281.

[0189] In this context, the fragment may be an N-terminal truncated portion or a C-terminal truncated portion of SEQ ID NO: 280 or SEQ ID NO: 281. In embodiments, the immunogenic fragments of SEQ ID NO: 280 or SEQ ID NO: 281 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 280 or SEQ ID NO: 281 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0190] MAGE-A3 amino acid and nucleic acid sequences

[0191] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of MAGE-A3 tumor neoantigens and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 282 or SEQ ID NO: 283. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of MAGE-A3 tumor neoantigens, or an immunogenic fragment or variant thereof, wherein said antigenic peptide comprises or constitutes at least one amino acid sequence, said amino acid sequence being identical to or having at least 80% identity with any of SEQ ID NO: 282 or SEQ ID NO: 283.

[0192] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 282 or SEQ ID NO: 283, or a C-terminal truncation of SEQ ID NO: 282 or SEQ ID NO: 283. In embodiments, the immunogenic fragments of SEQ ID NO: 282 or SEQ ID NO: 283 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 282 or SEQ ID NO: 283 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids in size.

[0193] ATAD2 amino acid and nucleic acid sequence

[0194] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the ATAD2 tumor neoantigen and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or immunogenic variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 284 to SEQ ID NO: 286. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of the ATAD2 tumor neoantigen, or an immunogenic fragment or variant thereof, wherein said antigenic peptide comprises or constitutes at least one amino acid sequence, said amino acid sequence being identical to or having at least 80% identity with any of SEQ ID NO: 284 to SEQ ID NO: 286.

[0195] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 284 to SEQ ID NO: 286 or a C-terminal truncation of SEQ ID NO: 284 to SEQ ID NO: 286. In embodiments, the immunogenic fragments of SEQ ID NO: 284 to SEQ ID NO: 286 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 284 to SEQ ID NO: 286 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope. Typical immunogenic fragments in this context are 5 to 20 amino acids in size, or 8 to 15 amino acids.

[0196] BRAF amino acid and nucleic acid sequence

[0197] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of BRAF tumor neoantigens and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 262 to SEQ ID NO: 266. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of BRAF tumor neoantigens, said antigenic peptide comprising or constituting at least one amino acid sequence, or an immunogenic fragment or variant thereof, said amino acid sequence being identical to or having at least 80% identity with any of SEQ ID NO: 262 to SEQ ID NO: 266.

[0198] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 262 to SEQ ID NO: 266 or a C-terminal truncation of SEQ ID NO: 266. In embodiments, the immunogenic fragments of SEQ ID NO: 262 to SEQ ID NO: 266 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 262 to SEQ ID NO: 266 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope.

[0199] EGFR amino acid and nucleic acid sequence

[0200] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of EGFR tumor neoantigens and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 255 to SEQ ID NO: 257. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of EGFR tumor neoantigens, said antigenic peptide comprising or constituting at least one amino acid sequence, or an immunogenic fragment or variant thereof, said amino acid sequence being identical to or having at least 80% identity with any of SEQ ID NO: 255 to SEQ ID NO: 257.

[0201] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 255 to SEQ ID NO: 257 or a C-terminal truncation of SEQ ID NO: 257. In embodiments, the immunogenic fragments of SEQ ID NO: 255 to SEQ ID NO: 257 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 255 to SEQ ID NO: 257 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope.

[0202] TP53 amino acid and nucleic acid sequence

[0203] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the TP53 tumor neoantigen and comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 258 to SEQ ID NO: 261. Therefore, in a preferred embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins of the TP53 tumor neoantigen, wherein said antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, wherein the amino acid sequence is identical to or has at least 80% identity with any of SEQ ID NO: 258 to SEQ ID NO: 261.

[0204] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 258 to SEQ ID NO: 261 or a C-terminal truncation of SEQ ID NO: 258 to SEQ ID NO: 261. In embodiments, the immunogenic fragments of SEQ ID NO: 258 to SEQ ID NO: 261 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 258 to SEQ ID NO: 261 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope.

[0205] DIP2B, TRRAP, RAC1, AP4B1, VWA5A, CLTC, RPL9, RPAP1, RPE65, PATZ1, PIK3CA, NFE2L2, CNOT9 amino acid and nucleic acid sequence

[0206] In the embodiments, at least one antigenic peptide is selected from peptides or proteins of DIP2B, TRRAP, RAC1, AP4B1, VWA5A, CLTC, RPL9, RPAP1, RPE65, PATZ1, PIK3CA, NFE2L2, or CNOT9 tumor neoantigens, and comprises or constitutes at least one amino acid sequence, or any of these immunogenic fragments or immunogenic variants, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 287 to SEQ ID NO: 317. Therefore, in an embodiment, the artificial nucleic acid comprises at least one CDS encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from DIP2B, TRRAP, RAC1, AP4B1, VWA5A, CLTC, RPL9, RPAP1, RPE65, PATZ1 tumor neoantigens, or a peptide or protein thereof, said antigenic peptide comprising or constituting at least one amino acid sequence, or an immunogenic fragment or variant thereof thereof, said amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO: 287 to SEQ ID NO: 317.

[0207] In this context, the fragment may be an N-terminal truncation of SEQ ID NO: 287 to SEQ ID NO: 317 or a C-terminal truncation of SEQ ID NO: 287 to SEQ ID NO: 317. In embodiments, the immunogenic fragments of SEQ ID NO: 287 to SEQ ID NO: 317 are truncated by 1, 2, 3, or 4 amino acids at the N-terminus and / or truncated by 1, 2, 3, or 4 amino acids at the C-terminus. The immunogenic fragments of SEQ ID NO: 287 to SEQ ID NO: 317 still contain at least one epitope capable of evoking an immune response, preferably a T-cell epitope.

[0208] Preferred nucleic acid coding sequences are described below, which encode antigenic peptides as defined herein as tumor neoantigens, and these nucleic acid coding sequences may be included in at least one coding sequence of the artificial nucleic acid of the present invention.

[0209] In a preferred embodiment, at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from tumor neoantigens, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants. In this context, preferably, the fragment is at least 24 nucleotides in length.

[0210] In a preferred embodiment, at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins of tumor neoantigens, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants. In this context, preferably, the fragment is at least 24 nucleotides in length.

[0211] Suitably, the nucleic acid sequence encoding at least one antigenic peptide from a tumor neoantigen is a codon-modified CDS, preferably wherein the codon-modified CDS is selected from C-maximized CDS (as defined herein), CAI-maximized CDS (as defined herein), human codon selection adapted CDS (as defined herein), G / C content modified CDS (as defined herein), and G / C optimized CDS (as defined herein), or any combination thereof. In a preferred embodiment within this context, at least one codon-modified CDS is a G / C optimized CDS.

[0212] Therefore, in a particularly preferred embodiment, at least one coding sequence comprises a GC-optimized nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins of tumor neoantigens, wherein the nucleic acid sequence is identical to or has at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 449 to SEQ ID NO: 511. In this context, preferably, the fragment is at least 24 nucleotides in length.

[0213] Preferred amino acid and nucleic acid sequences in this context are provided in Table 2. Each row corresponds to an antigenic peptide of a suitable tumor neoantigen. Column A provides a brief description of the corresponding tumor neoantigen. Column B provides the SEQ ID NO of the protein (amino acid sequence) of the preferred antigenic peptide of the tumor neoantigen. Column C provides the SEQ ID NO of the corresponding G / C-optimized nucleic acid sequence encoding the preferred antigenic peptide of the tumor neoantigen. Column F shows the preferred cancer indications for the antigen (N = NSCLC; M = melanoma).

[0214] Table 2: Preferred antigens for tumor neoantigens (amino acid sequences and coding sequences)

[0215]

[0216] In a particularly preferred embodiment, at least one antigenic peptide comprises or constitutes at least one amino acid sequence selected from column B of Table 2, or any of these immunogenic fragments. In an even more preferred embodiment, at least one antigenic peptide comprises or constitutes at least one amino acid sequence selected from rows 1 to 11 of column B of Table 2, or any of these immunogenic fragments.

[0217] In a particularly preferred embodiment, at least one coding sequence of the artificial nucleic acid of the present invention comprises or constitutes at least one nucleic acid sequence provided in column C of Table 2. In an even more preferred embodiment, at least one coding sequence of the artificial nucleic acid of the present invention comprises or constitutes at least one nucleic acid sequence, or an immunogenic fragment thereof, provided in rows 1 to 11 of column C of Table 2.

[0218] Other peptide or protein characteristics

[0219] In the implementation scheme, at least one coding sequence encodes 2 to 10 different tumor antigens as specified herein, preferably 2 to 5 different tumor antigens as specified herein. Therefore, the coding sequence can encode multiple tumor antigens provided in the form of a fusion construct.

[0220] Appropriately, different tumor antigens are selected to achieve increased coverage in specific cancer diseases, such as NSCLC, HNSCC, or melanoma. Appropriately, the coverage values ​​of all different tumor antigens are greater than 0.5, preferably greater than 0.6, and more preferably greater than 0.7.

[0221] In the context of this invention, "different tumor antigens" refers to differences at the amino acid sequence level and should be understood as differences at at least one amino acid position or different amino acid sequence lengths. Therefore, "different tumor antigens" can originate from the same neotumor tumor, but the "different tumor antigens" should differ at at least one amino acid position or length.

[0222] Therefore, at least one coding sequence may encode 2 to 10 different tumor antigens, preferably 2 to 5 different tumor antigens, wherein said different tumor antigens are selected from peptides or proteins encoded by long non-coding RNA (lncRNA) as defined herein, and / or peptides or proteins selected from neotumor antigens as defined herein, or any immunogenic fragments or variants of these. Suitable biological amino acid and nucleic acid sequences in this context are as defined above, and may in particular be selected from Tables 1A and 1B and Table 2.

[0223] In this preferred embodiment within the context, different tumor antigens are selected to cover different HLA types. Thus, the different tumor antigens are selected from at least one tumor antigen as defined herein that contains at least one epitope binding to HLA-A type, at least one tumor antigen as defined herein that contains at least one epitope binding to HLA-B type, and at least one tumor antigen as defined herein that contains at least one epitope binding to HLA-C type.

[0224] In the implementation, different tumor antigens are separated by adapter elements. Therefore, the coding sequence can encode multiple different tumor antigens provided in the form of a fusion construct, wherein the different tumor antigens in the fusion construct are separated by adapter elements. Suitable adapter elements may be selected from SEQ ID NO: 2937, SEQ ID NO: 76400 to SEQ ID NO: 76418, and SEQ ID NO: 77018 to 77058 of published PCT patent application WO2019008001.

[0225] In an embodiment, at least one CDS encodes at least one additional amino acid sequence selected from at least one immune response activation signal transducer, preferably from at least one transmembrane domain and / or at least one cytoplasmic domain of the at least one immune response activation signal transducer.

[0226] Preferably, the immune response activation signal transduction protein is located on the outer side of the plasma membrane and is readily and repeatedly internalized into the endosome pathway, preferably targeting the encoded and translated antigen to the desired intracellular pathway, which preferably intersects with the MHC class I pathway and, in particular, with the MHC class II pathway, to efficiently deliver the antigen sequence to the MHC class I and MHC class II processing compartments (as described in WO2019008001, the entire contents of which are incorporated herein by reference).

[0227] In a preferred embodiment, the immune response activation signal transduction protein is selected from CTLA4.

[0228] In a preferred embodiment, at least one immune response activation signal transduction protein comprises or consists of at least one amino acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the sequences in published PCT patent application WO2019008001, or any fragments or variants thereof.

[0229] In the embodiments, at least one coding sequence encodes at least one additional amino acid sequence selected from at least one T helper cell epitope. Suitable T helper cell epitopes may be selected from SEQ ID NO: 3083 to SEQ ID NO: 3294 of published PCT patent application WO2019008001.

[0230] In an embodiment, at least one coding sequence encodes at least one additional amino acid sequence selected from at least one signal peptide. Suitable signal peptides may be selected from SEQ ID NO: 1 to SEQ ID NO: 156 and SEQ ID NO: 76948 to SEQ ID NO: 76951 of published PCT patent application WO2019008001. In a preferred embodiment, at least one signal peptide is selected from CTLA4.

[0231] Nucleic acid sequence characteristics and implementation plan

[0232] Suitable features and embodiments of artificial nucleic acid sequences relating to the present invention (e.g., nucleic acid type, nucleic acid structure, nucleic acid element, nucleic acid modification, etc.) are provided and described in detail below. It is worth noting that the features defining the nucleic acid sequence can also be applied to any nucleic acid sequence in any other aspect of the present invention (e.g., compositions, combinations, kits, medical uses).

[0233] Suitable encoding sequence:

[0234] According to a preferred embodiment, the artificial nucleic acid of the present invention comprises at least one coding sequence that encodes at least one tumor antigen as defined herein. In this context, any coding sequence encoding at least one tumor antigen as defined herein, or fragments and variants thereof, can be understood as a suitable coding sequence and therefore can be included in the nucleic acid of the present invention.

[0235] In a preferred embodiment, the artificial nucleic acid is a modified and / or stabilized nucleic acid.

[0236] According to the preferred embodiment, artificial nucleic acids can therefore be provided as “stable nucleic acids,” i.e., nucleic acids exhibiting improved in vivo degradation resistance, and / or nucleic acids exhibiting improved in vivo stability, and / or nucleic acids exhibiting improved in vivo translatability. This is particularly important in embodiments where the nucleic acid is RNA.

[0237] Preferably, the artificial nucleic acid of the present invention can be provided as a "stable nucleic acid", more preferably a "stable RNA".

[0238] The following text describes suitable modifications / adaptations that can "stabilize" nucleic acids and optimize RNA.

[0239] In a particularly preferred embodiment, the artificial nucleic acid comprises at least one codon-modified coding sequence (CDS). Suitably, the amino acid sequence encoded by the at least one codon-modified coding sequence is unmodified compared to the amino acid sequence encoded by the corresponding wild-type or reference coding sequence.

[0240] The term "codon-modified coding sequence" refers to a coding sequence that differs from the corresponding wild-type or reference coding sequence at at least one codon (a nucleotide triplet encoding one amino acid). Appropriately, in the context of this invention, codon-modified coding sequences can exhibit improved in vivo degradation resistance, and / or improved in vivo stability, and / or improved in vivo translatability. Codon modification in its broadest sense utilizes the degeneracy of the genetic code, where multiple codons can encode the same amino acid and can be used interchangeably to optimize / modify the coding sequence for in vivo applications.

[0241] In the implementation, at least one code sequence (CD) is a codon-modified CD, wherein the codon-modified CD is selected from C-maximized coding sequences (as further defined in WO2021239880 [page 122, lines 33-39], which is incorporated herein by reference); CAI-maximized coding sequences (as further defined in WO2021239880 [page 123, lines 33-44], which is incorporated herein by reference); human codon selection adapted CDs (as further defined in WO2021239880 [page 123, lines 7-17], which is incorporated herein by reference); G / C content modified coding sequences (as further defined in WO2021239880 [page 123, lines 19-31], which is incorporated herein by reference); G / C optimized coding sequences; or any combination thereof.

[0242] When transfected into mammalian host cells, the nucleic acid containing the codon-modified coding sequence has stability for 12 to 18 hours, or more than 18 hours, such as 24 hours, 36 hours, 48 ​​hours, 60 hours, 72 hours, or more than 72 hours, and is able to be expressed by the mammalian host cells.

[0243] When transfected into mammalian host cells, an artificial nucleic acid containing a codon-modified coding sequence is translated into a protein, wherein the amount of said protein is at least equal to, or preferably at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 100%, or at least 200%, or more than 200% of the amount of protein obtained by transfecting into a naturally occurring coding sequence, wild-type coding sequence, or reference coding sequence in a mammalian host cell.

[0244] In a particularly preferred embodiment, the artificial nucleic acid may be modified such that the G / C content of at least one coding sequence is optimized compared to the G / C content of a corresponding wild-type or reference coding sequence (“G / C-optimized coding sequence”). In this context, “optimization” means that the G / C content of the coding sequence is preferably increased to substantially the highest possible G / C content. The generation of the G / C-optimized nucleic acid sequence can be performed using the method according to WO2002098443. In this context, the disclosure of WO2002098443 is included in the present invention in its entirety.

[0245] In a preferred embodiment, at least one codon-modified coding sequence is a G / C-optimized coding sequence. Such G / C-optimized nucleic acid sequences are provided in Table 1A, Table 1B, or Table 2.

[0246] In a preferred embodiment, at least one coding sequence of the artificial nucleic acid, preferably RNA, has a G / C content of at least about 50%, 55%, or 60%. In a particular embodiment, at least one coding sequence of the nucleic acid has a G / C content of at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, or 70%. In a particularly preferred embodiment, the artificial nucleic acid may be modified such that the G / C content of at least one CDS is optimized compared to the G / C content of a corresponding wild-type or reference CDS (“G / C-optimized coding sequence”). In this context, “optimization” means that the G / C content of the coding sequence is preferably increased to substantially the highest possible G / C content. The generation of a G / C-optimized nucleic acid sequence can be performed using the method according to WO2002098443. In this context, the disclosure of WO2002098443 is included in the present invention in its entirety.

[0247] UTR:

[0248] In a preferred embodiment, the artificial nucleic acid of the present invention, preferably the RNA, comprises at least one untranslated region (UTR) or UTR element.

[0249] The terms "untranslated region," "UTR," or "UTR element" refer to a portion of a nucleic acid molecule that is typically located at the 5' or 3' end of a coding sequence. The UTR is not translated into a protein. The UTR can be a part of a nucleic acid, such as RNA. The UTR can contain elements used to control gene expression, also known as regulatory elements. Such regulatory elements can be, for example, ribosome binding sites, miRNA binding sites, promoter elements, etc.

[0250] In a preferred embodiment, the artificial nucleic acid comprises a coding sequence and a 5'-UTR and / or a 3'-UTR. Notably, the UTR may contain regulatory sequence elements that determine RNA renewal, stability, and localization. Furthermore, the UTR may contain sequence elements that enhance translation. In medical applications, the translation of nucleic acids into at least one peptide or protein is crucial for therapeutic efficacy. Certain combinations of 3'-UTRs and / or 5'-UTRs can enhance the expression of operablely linked coding sequences encoding peptides or proteins as defined herein. Nucleic acid molecules containing such UTR combinations are advantageously capable of rapidly and transiently expressing encoded tumor antigens upon administration to a subject.

[0251] Suitable, the artificial nucleic acid comprises at least one untranslated region (UTR), preferably selected from at least one 5'-UTR and / or at least one 3'-UTR. The 5'-UTR or the 3'-UTR may be derived from a naturally occurring gene or may be synthetically engineered. In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence as defined herein, operably linked to at least one 3'-UTR and / or at least one 5'-UTR.

[0252] Preferably, at least one untranslated region (UTR) is selected from at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR.

[0253] As used in this article, the term "heterologous" sequence or "heterologous" UTR refers to a nucleic acid sequence or UTR that is not derived from the same gene, the same genome fusion, or the same naturally occurring transcript (e.g., lncRNA). Therefore, heterologous sequences or UTRs can be derived from the same organism (e.g., human) or different organisms. Heterologous sequences or heterologous UTRs do not occur in the same nucleic acid in nature.

[0254] In a preferred embodiment, the artificial nucleic acid of the present invention contains at least one 3'-UTR.

[0255] The term "3'-untranslated region" or "3'-UTR" refers to the nucleic acid portion located at the 3' (downstream) of the CDS that is not translated into protein. The 3'-UTR can be a nucleic acid portion located between the CDS and (optionally) the terminal poly(A) sequence. The 3'-UTR can contain elements used to control gene expression, also known as regulatory elements. Such regulatory elements can be, for example, ribosome binding sites or miRNA binding sites.

[0256] Preferably, the artificial nucleic acid contains at least one 3'-UTR, which may be derived from a gene associated with RNA having an enhanced half-life (i.e., a gene that provides stable RNA).

[0257] In some implementations, the 3'-UTR includes one or more of the following: a polyadenylation signal, a protein binding site that affects the stability of the intracellular nucleic acid position, or one or more miRNAs or miRNA binding sites.

[0258] In a preferred embodiment, the artificial nucleic acid comprises at least one 3'-UTR, wherein the at least one 3'-UTR comprises or constitutes a nucleic acid sequence derived from or selected from the 3'-UTR of a gene, or a homolog, fragment, or variant derived from or selected from any of these genes, said gene being selected from: PSMB3, ALB7, α-globin, β-globin, ANXA4, CASP1, COX6B1, FIG4, GNAS, NDUFA1, RPS9, SLC7A3, or TUBB4B.

[0259] In a preferred embodiment, at least one 3'-UTR derived from or selected from PSMB3, ALB7, α-globin, β-globin, ANXA4, CASP1, COX6B1, FIG4, GNAS, NDUFA1, RPS9, SLC7A3, or TUBB4B comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 66 to SEQ ID NO: 95, SEQ ID NO: 112 to SEQ ID NO: 123, or a fragment or variant thereof.

[0260] In other embodiments, at least one 3'-UTR comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the fragments or variants of SEQ ID NO: 96 to 111.

[0261] In a particularly preferred embodiment, the artificial nucleic acid comprises a 3'-UTR derived from or selected from the PSMB3 gene, wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the fragment or variant of SEQ ID NO: 66, SEQ ID NO: 67, or any of these.

[0262] In a preferred embodiment, the artificial nucleic acid of the present invention comprises at least one 5'-UTR.

[0263] The term "5'-untranslated region" or "5'-UTR" refers to the nucleotide portion located at the 5' (i.e., "upstream") of the cytokine sequence (CDS) that is not translated into protein. The 5'-UTR can be a portion of the nucleotide at the 5' of the CDS. Typically, the 5'-UTR begins at the transcription start site and terminates before the start codon of the CDS. The 5'-UTR can contain elements used to control gene expression, also known as regulatory elements. Such regulatory elements can be, for example, ribosome binding sites, miRNA binding sites, etc. The 5'-UTR can be modified, for example, by adding a 5'-cap structure (e.g., as defined for mRNA below) through enzymatic or co-transcriptional processes.

[0264] Preferably, the artificial nucleic acid contains at least one 5'-UTR, which may be derived from a gene associated with RNA having an enhanced half-life (i.e., a gene that provides stable RNA).

[0265] In some implementations, the 5'-UTR includes one or more of the following: a protein binding site that affects the stability of the nucleic acid position in the cell, or one or more miRNAs or miRNA binding sites (as defined above).

[0266] Therefore, miRNAs or miRNA binding sites can be removed or introduced from the 5'-UTR to adapt nucleic acid expression to the desired cell type or tissue (e.g., muscle cells).

[0267] In a preferred embodiment, the artificial nucleic acid comprises at least one 5'-UTR, wherein the at least one 5'-UTR comprises a nucleic acid sequence derived from or selected from the 5'-UTR of a gene, or a homolog, fragment, or variant derived from or selected from any of these genes, wherein the gene is selected from: HSD17B4, RPL32, AIG1, α-globin, ASAH1, ATP5A1, COX6C, DPYSL2, MDR, MP68, NDUFA4, NOSIP, RPL31, RPL35A, SLC7A3, TUBB4B, or UBQLN2.

[0268] In a preferred embodiment, at least one 5'-UTR derived from or selected from HSD17B4, RPL32, AIG1, α-globin, ASAH1, ATP5A1, COX6C, DPYSL2, MDR, MP68, NDUFA4, NOSIP, RPL31, RPL35A, SLC7A3, TUBB4B, or UBQLN2 contains or constitutes a fragment or variant identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants.

[0269] In other embodiments, at least one 5'-UTR comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the fragments or variants of SEQ ID NO: 46 to SEQ ID NO: 63.

[0270] In a particularly preferred embodiment, the nucleic acid comprises a 5'-UTR derived from or selected from the HSD17B4 gene, wherein at least one heterologous 5'-UTR comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the fragments or variants of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 64, SEQ ID NO: 65, preferably SEQ ID NO: 13, or any of these.

[0271] In the implementation, at least one nucleic acid, preferably RNA, comprises at least one coding sequence operatively linked to a 3'-UTR element and / or a 5'-UTR element selected from the 5'-UTR / 3'-UTR combination provided in WO2021239880 [page 127, line 35 to page 128, line 2], which is incorporated herein by reference.

[0272] In a preferred embodiment, at least one heterogeneous 5'-UTR is selected from HSD17B4 and the at least one heterogeneous 3' UTR is selected from PSMB3.

[0273] Therefore, in a particularly preferred embodiment, the artificial nucleic acid, preferably the RNA, comprises at least one cds as defined herein, encoding at least one tumor antigen as defined herein, wherein the cds is operatively linked to the HSD17B4 5'-UTR and the PSMB3 3'-UTR (HSD17B4 / PSMB3(a-1)). The inventors have demonstrated that this embodiment is particularly advantageous for expressing tumor antigens in human cells.

[0274] In various implementations, the nucleic acid, such as the RNA, is monocistronic, bicistronic, or polycistronic.

[0275] In a preferred embodiment, the nucleic acid, preferably the RNA, is monocistronic.

[0276] Therefore, in a particularly preferred embodiment, the artificial nucleic acid contains a ribosome binding site, also known as a "Kozak sequence," which is identical or has at least 80%, 85%, 90%, or 95% identity with any sequence in SEQ ID NO: 1 or SEQ ID NO: 2, or sequences GCCGCCACC(DNA), GCCGCCACC(RNA), GCCACC(DNA), GCCACC(RNA), ACC(DNA), or ACC(RNA), or any fragments or variants thereof. In a preferred embodiment, the "Kozak sequence" comprises or constitutes an RNA sequence ACC.

[0277] DNA and RNA constructs:

[0278] In a particularly preferred embodiment, the artificial nucleic acid of the present invention is an isolated nucleic acid. The term "isolated nucleic acid" does not include the cell or object containing the nucleic acid, but refers to an artificial nucleic acid as an isolated molecule or a collection of isolated molecules. For example, "isolated nucleic acid" may be an artificial nucleic acid isolated or purified from cells (e.g., cell cultures, bacterial cultures), or it may be an artificial nucleic acid (e.g., RNA) isolated from RNA through in vitro transcription.

[0279] In a particularly preferred embodiment, the artificial nucleic acid of the present invention is a therapeutic nucleic acid.

[0280] Therefore, artificial nucleic acids are appropriately used in a therapeutic context, particularly to provide a therapeutic approach for providing at least one tumor antigen according to the present invention.

[0281] In the implementation scheme, the artificial nucleic acid of the present invention is selected from DNA or RNA.

[0282] In the embodiments, the artificial nucleic acid of the present invention is selected from DNA. The DNA can be any type of DNA containing cds as defined herein, including any type of single-stranded DNA, double-stranded DNA, linear DNA, and circular DNA. Suitable DNA in the context of the present invention can be selected from bacterial plasmids, adenoviruses, poxviruses, parapoxoviruses (ORF viruses), vaccinia virus, fowlpox virus, herpesviruses, adeno-associated virus (AAV), alphaviruses, lentiviruses, λ phages, lymphocytic choriomeningitis virus, and Listeria and Salmonella species. In a preferred embodiment, the DNA is viral DNA, preferably AAV DNA.

[0283] In a particularly preferred embodiment, the artificial nucleic acid of the present invention is RNA. The RNA can be any type of RNA containing cds as defined herein, including any type of single-stranded RNA, double-stranded RNA, linear RNA, and circular RNA. In a preferred embodiment, the RNA can be any type of RNA containing cds as defined herein, including any type of single-stranded RNA, double-stranded RNA, linear RNA, and circular RNA, wherein said RNA is not lncRNA. In a preferred embodiment, the RNA is selected from mRNA, circular RNA, replicon RNA, or self-replicating RNA, or viral RNA, preferably mRNA or circular RNA.

[0284] In the implementation scheme, the RNA is not a lncRNA. Specifically, the artificial nucleic acid of the present invention is not selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, which means that the RNA of the present invention can undoubtedly contain lncRNA encoding lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9, etc. The coding sequences of peptides encoded by 1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25 are different in sequence and length from those encoded by lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25.

[0285] In the implementation, the RNA is circular RNA. "Circular RNAs" (circRNAs) are RNAs linked to form a loop and therefore do not contain a 3' or 5' end. The circRNA contains at least one cds as defined herein. circRNA construct designs can be obtained from claims 1 to 51 of WO2023073228, which are incorporated herein by reference.

[0286] In other embodiments, the RNA is a replicon RNA or a self-replicating RNA. Such constructs may encode replicase elements from, for example, alphaviruses (e.g., SFV, SIN, VEE, or RRV) and coding sequences as defined herein.

[0287] In a particularly preferred embodiment, the RNA is selected from mRNA. Therefore, the artificial nucleic acid of the present invention is mRNA, preferably isolated mRNA.

[0288] In the context of this invention, mRNA is preferred for providing the tumor antigen according to the invention because mRNA allows for dose regulation, transient expression, complete degradation of mRNA after protein synthesis, and does not pose a risk of insertional mutations.

[0289] Preferably, the artificial nucleic acid, more preferably the RNA, comprises about 50 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 about 2,000 to about 5,000 nucleotides.

[0290] Poly(N) sequence, histone stem-loop:

[0291] In a preferred embodiment, the artificial nucleic acid comprises at least one poly(N) sequence, such as at least one poly(A) sequence, at least one poly(U) sequence, at least one poly(C) sequence, or a combination thereof.

[0292] In a preferred embodiment, the artificial nucleic acid, such as the RNA, comprises at least one poly(A) sequence. In some embodiments, the artificial nucleic acid comprises at least two, three, or more than three poly(A) sequences.

[0293] The term "poly(A) sequence" or "poly(A) tail" refers to a sequence of up to 1000 adenosines typically located at the 3' end of a linear RNA. Typically, poly(A) sequences are homopolymeric. Alternatively, a poly(A) sequence can be interrupted by at least one nucleotide different from adenosine.

[0294] In embodiments, at least one poly(A) sequence may comprise about 20 to about 500 adenosine nucleotides, about 40 to about 250 adenosine nucleotides, about 60 to about 250 adenosine nucleotides, preferably about 60 to about 150 adenosine nucleotides. Suitably, the length of the poly(A) sequence may be at least about or even more than about 50, 64, 75, 100, 200, 300, 400 or 500 adenosine nucleotides, preferably continuous adenosine nucleotides.

[0295] In a particularly preferred embodiment, at least one poly(A) sequence comprises about 100 adenosine nucleotides (A100), preferably about 100 consecutive adenosine nucleotides.

[0296] In an alternative embodiment, at least one nucleic acid comprises at least one interrupted poly(A) sequence, wherein the poly(A) sequence is interrupted by non-adenosine nucleotides, preferably by about 10 non-adenosine (N10) nucleotides. In this context, the preferred poly(A) sequence is A30-N10-A70.

[0297] As defined herein, a poly(A) sequence can be located directly at the 3' end of an artificial nucleic acid, preferably RNA. In a preferred embodiment, the 3' terminal nucleotide (i.e., the last 3' terminal nucleotide in the polynucleotide chain) is at least one 3' terminal A nucleotide of a poly(A) sequence. The term "directly at the 3' end" should be understood as exactly at the 3' end—in other words, the 3' end of the nucleic acid consists of an A-terminated poly(A) sequence.

[0298] Advantageously, the presence of a 3' end ending in an adenosine nucleotide reduces the likelihood of RNA-induced interferon, such as IFNα, from the present invention when administered to humans, for example, as a drug. This is important because induction of interferon, such as IFNα, is considered one of the major factors inducing side effects.

[0299] Therefore, in a particularly preferred embodiment, the artificial nucleic acid of the present invention, such as RNA, comprises a poly(A) sequence of about 100 consecutive adenosine nucleotides, wherein the poly(A) sequence is located directly at the 3' end of the RNA, and optionally wherein the 3' terminal nucleotide is adenosine.

[0300] In a preferred embodiment, the artificial nucleic acid, such as the RNA, comprises at least one histone stem-loop (hSL) or histone stem-loop structure. In the context of this invention, the hSL may be located in the 3' region. The term "histone stem-loop" (hSL) refers to a nucleic acid sequence that forms a stem-loop secondary structure and is primarily found in histone mRNA. Histone stem-loop sequences / structures may suitably be selected from the hSL sequences disclosed in WO2012019780. The hSL sequences used herein may be derived from formula (I) or (II) of WO2012019780. Therefore, an artificial nucleic acid may comprise at least one hSL sequence derived from a specific formula (Ia) or formula (IIa) of WO2012019780.

[0301] In a preferred embodiment, at least one histone stem-loop sequence comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the fragments or variants of SEQ ID NO: 3 or SEQ ID NO: 4. Preferably, the histone stem-loop sequence comprises or constitutes a nucleic acid sequence according to SEQ ID NO: 4, or a fragment thereof.

[0302] In a preferred embodiment, the artificial nucleic acid comprises a 3' terminal sequence element. The 3' terminal sequence element represents the 3' end of the RNA. A 3' terminal sequence element may comprise at least one poly(N) sequence as defined herein, and optionally, at least one hSL as defined herein.

[0303] In a preferred embodiment, the artificial nucleic acid comprises at least one 3' end sequence element, said element comprising or constituting an RNA sequence that is identical to or has at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 5 to SEQ ID NO: 11, or fragments or variants thereof.

[0304] In a preferred embodiment, the artificial nucleic acid comprises a 3' end sequence element containing hSL as defined herein, followed by a poly(A) sequence containing approximately 100 consecutive adenosines.

[0305] In a particularly preferred embodiment, the artificial nucleic acid comprises a 3' end sequence element comprising or constituting a nucleic acid sequence that is identical to or has at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 5 or SEQ ID NO: 6, or a fragment or variant thereof.

[0306] In some embodiments, the artificial nucleic acid includes a 5' terminal sequence element that comprises or constitutes a nucleic acid sequence identical to or having at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of the sequences AGGAGA, GGGAGA, GGGAAA, AGAAUA, AGAUUA, GAUGGG, or GGGCG, or a fragment or variant of these sequences, preferably AGGAGA. Such a 5' terminal sequence element may include, for example, a binding site for T7 RNA polymerase. Furthermore, the first nucleotide of the 5' terminal start sequence may preferably contain a 2'O methylation, such as 2'O methylated guanosine or 2'O methylated adenosine.

[0307] Modified nucleotides:

[0308] In various implementation schemes, the artificial nucleic acid, preferably RNA, is modified, wherein the modification refers to chemical modification, including backbone modification and sugar or base modification.

[0309] Modified nucleic acids or RNA may contain nucleoside analogs / modifications, such as backbone modifications, sugar modifications, or base modifications. In the context of this invention, backbone modification refers to the chemical modification of the backbone phosphate group of an RNA nucleotide. Sugar modification in the context of this invention refers to the chemical modification of the sugar content of an RNA nucleotide. Furthermore, base modification in the context of this invention refers to the chemical modification of the base portion of an RNA nucleotide. In this context, nucleoside analogs or modifications are preferably selected from nucleoside analogs suitable for transcription and / or translation.

[0310] Therefore, in a preferred embodiment, the nucleic acid of the present invention, preferably the RNA, comprises at least one modified nucleotide.

[0311] In some embodiments, at least one modified nucleotide is selected from pseudouridine (ψ), N1-methylpseudouridine (m1ψ), N1-ethylpseudouridine, 2-thiouridine, 4'-thiouridine, 5-methylcytidine, 5-methyluridine, 2-thio-1-methyl-1-denitro-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine, and 2'-O-methyluridine.

[0312] In this context, pseudouridine (ψ) and N1-methylpseudouridine (m1ψ) are particularly preferred. Therefore, in a preferred embodiment, the nucleic acid is RNA containing at least one modified nucleotide, preferably a modified nucleotide selected from pseudouridine (ψ) or N1-methylpseudouridine (m1ψ).

[0313] In some implementations, uracil in substantially all, for example substantially 100% of the coding sequence (or complete nucleic acid sequence) is chemically modified, preferably chemically modified at the 5-position of uracil.

[0314] In a preferred embodiment, 100% of the uracil in the intact nucleic acid sequence, preferably the RNA sequence, is replaced by a modified nucleotide, preferably N1-methylpseudouridine (m1ψ). Alternatively, 100% of the uracil in the intact nucleic acid sequence, preferably the RNA sequence, is replaced by pseudouridine (ψ).

[0315] In a preferred embodiment, the artificial nucleic acid, preferably RNA, does not contain chemically modified nucleotides. It is noteworthy that a 5'-cap structure, as defined below, is generally not considered a chemically modified nucleotide. Therefore, the artificial nucleic acid, preferably RNA, comprises a sequence consisting only of G, C, A, and U nucleotides, and thus does not contain modified nucleotides, and optionally includes a 5'-cap structure. RNA constructs that do not contain chemically modified nucleotides may be beneficial in the context of cancer therapy, as providing cancer antigens via unmodified RNA (compared to RNA modified with m1ψ or ψ) can induce a stronger T-cell response.

[0316] In a preferred embodiment, the artificial nucleic acid, preferably the RNA of the present invention, does not contain an N1-methylpseudouridine (m1Ψ) substitution site or a pseudouridine (ψ) substitution site.

[0317] Hat structure:

[0318] In a preferred embodiment, the artificial nucleic acid is RNA containing a 5'-cap structure.

[0319] Such a 5'-cap structure appropriately stabilizes nucleic acids and / or enhances the expression of encoded tumor antigens and / or reduces stimulation of the innate immune system after administration.

[0320] Therefore, in a preferred embodiment, the artificial nucleic acid, preferably RNA, comprises a 5'-cap structure, preferably m7G, cap0, cap1, cap2, modified cap0, or modified cap1 structure.

[0321] The term "5'-cap structure" as used herein will be recognized and understood by those skilled in the art, and is intended, for example, to refer to a 5'-modified nucleotide, particularly a guanine nucleotide, located at the 5' end of RNA. Preferably, the 5'-cap structure is linked to RNA via a 5'-5'-triphosphate linker.

[0322] Suitable 5'-cap structures in the context of this invention include cap0 (methylation of the first nucleobase, e.g., m7GpppN), cap1 (additional methylation of the ribose of the nucleotide adjacent to m7GGpppN), 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 analogs), modified ARCA (e.g., phosphate thioester modified ARCA), inosine, N1-methyl-guanosine, 2'-fluoro-guanosine, 7-denitro-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

[0323] Suitable, 5'-cap (cap0 or cap1) structures can be formed using cap analogs in chemical RNA synthesis or in vitro RNA transcription (co-transcription with capping).

[0324] As used herein, the term "cap analogue" will be recognized and understood by those skilled in the art, and is intended, for example, to refer to a non-polymerizable dinucleotide or trinucleotide that, when incorporated into the 5' end of a nucleic acid molecule, facilitates translation or localization and / or prevents degradation of the RNA molecule. Non-polymerizable means that the cap analogue will only be incorporated into the 5' end because it lacks a 5' triphosphate and therefore cannot be extended in the 3' direction by template-dependent polymerases, particularly template-dependent RNA polymerases.

[0325] In the implementation, cap1 structures are generated using cap analogs disclosed in WO2017053297, WO2017066793, WO2017066781, WO2017066791, WO2017066789, WO2017066782, WO2018075827, WO2017066797, and WO2023007019. Preferably, cap structures derived from the structures disclosed in claims 1 to 5 of WO2017053297 can be suitably used for co-transcriptionalization to generate the cap1 structure. Furthermore, any cap structure defined in claims 1 to 37 of WO2023007019 can be suitably used to generate cap1.

[0326] In a preferred embodiment, the 5'-cap structure may be co-transcribed and added using a trinucleotide cap analog as defined herein, preferably in an in vitro RNA transcription reaction as defined herein.

[0327] In a particularly preferred embodiment, the artificial nucleic acid, preferably the RNA of the present invention, comprises a cap1 structure or a modified cap1 structure.

[0328] In a preferred embodiment, the cap1 structure is formed by co-transcriptional capping using the trinucleotide cap analogs m7G(5')ppp(5')(2'OMeA)pG or m7G(5')ppp(5')(2'OMeG)pG. In this context, the particularly preferred cap1 analog is m7G(5')ppp(5')(2'OMeA)pG.

[0329] In other preferred embodiments, the cap1 structure is a modified cap1 structure and is formed by co-transcriptional capping using the trinucleotide cap analog 3'OMe-m7G(5')ppp(5')(2'OMeA)pG.

[0330] Alternatively, the 5'-cap structure can be enzymatically capped using a capping enzyme (e.g., vaccinia virus capping enzyme and / or 2'-O methyltransferase) to generate cap0, cap1, or cap2 structures.

[0331] Preferably, at least 80%, 85%, 90%, or 95% of the RNA species contain a cap structure, preferably a cap1 structure, as determined by a capping assay (e.g., by the assay described in claims 27 to 46 of WO2015101416).

[0332] Other RNA characteristics:

[0333] In various embodiments, an artificial nucleic acid is an RNA that provides at least one coding sequence that encodes at least one tumor antigen as defined herein, generated upon application to a cell or object.

[0334] The RNA of the present invention preferably includes suitable elements such as, for example, a 5' cap structure as defined herein, a 5' UTR as defined herein, a 3' UTR as defined herein, an hSL as defined herein, a poly(A) sequence as defined herein, and optional chemical modifications as defined herein.

[0335] In a preferred embodiment, the RNA is preferably in vitro transcribed RNA (e.g., in vitro transcribed mRNA).

[0336] In a preferred embodiment, the nucleotide mixture used for the in vitro transcription reaction of RNA (i.e., the proportion of each nucleotide in the mixture) is optimized for a given RNA sequence, preferably as described in WO2015188933. Therefore, in a preferred embodiment, the nucleic acid of the present invention is in vitro transcribed RNA, preferably wherein the in vitro transcription of RNA is performed in the presence of a sequence-optimized nucleotide mixture.

[0337] In a preferred embodiment, the artificial nucleic acid of the present invention is purified RNA, preferably purified mRNA. Suitably, the RNA of the present invention has been purified by at least one purification step.

[0338] The term "purified RNA" refers to RNA that has undergone certain purification steps to achieve a purity higher than that of the starting material (e.g., crude IVTRNA). Typical impurities include peptides, proteins, spermidine, BSA, aborted RNA fragments, dsRNA, free nucleotides, DNA, buffer components, etc. In this regard, it is desirable that the "RNA purity" be as close to 100% as possible. Preferably, the "purified RNA" used herein has a purity exceeding 75%, 80%, 85%, 90%, or 95%. Purity can be determined by analytical HPLC.

[0339] In the implementation scheme, the nucleic acid is purified RNA, which has been purified by at least one (RP)-HPLC, AEX, size exclusion chromatography (SEC), hydroxyapatite chromatography, tangential flow filtration (TFF), filtration, precipitation, core-bead flow through chromatography, oligo(dT) purification, cellulose-based purification, or any combination thereof. Preferably, the RNA has been purified using RP-HPLC (preferably as described in WO2008077592) and / or TFF (preferably as described in WO2016193206) and / or oligod(T) purification (preferably as described in WO2016180430) to, for example, remove dsRNA, uncapped RNA, and / or RNA fragments.

[0340] In the implementation scheme, the RNA has an integrity of at least 60%, 70%, 80%, and 90%. The term "RNA integrity" generally describes the presence of a complete RNA sequence. RNA integrity can be determined by RP-HPLC and can be based on determining the area under the expected full-length RNA peak in the chromatogram. In this regard, it is desirable that the "degree of RNA integrity" be as close to 100% as possible.

[0341] In a preferred embodiment, nucleic acid, preferably RNA, is used to treat or prevent diseases, disorders, or conditions, preferably tumors or cancerous diseases, disorders, or conditions, or any diseases, disorders, or conditions related to tumors or cancer. The term "cancer" according to this disclosure also includes cancer metastasis.

[0342] Preferred nucleic acid constructs:

[0343] In various implementation schemes, artificial nucleic acids comprise at least the following elements:

[0344] A) 5'-cap structure, preferably as specified herein;

[0345] B) Encoding at least one CDS of at least one tumor antigen as defined herein;

[0346] C) 5'-UTR and / or 3'-UTR, preferably as specified herein;

[0347] D) At least one poly(A) sequence, preferably as specified herein.

[0348] In a preferred embodiment, the artificial nucleic acid, preferably the RNA, comprises the following sequence elements, preferably in a 5' to 3' orientation:

[0349] A) 5'-cap structure, preferably cap1 structure;

[0350] B) 5'-UTR, preferably selected from or derived from the 5'-UTR of the HSD17B4 gene, or a fragment thereof;

[0351] C) Encoding a sequence that encodes at least one tumor antigen as defined herein;

[0352] D) 3'-UTR, preferably selected from or derived from the 3'-UTR of the PSMB3 gene, or a fragment thereof;

[0353] E) Optionally, histone stem-loop; and

[0354] F) poly(A) sequence, preferably containing about 100 A nucleotides.

[0355] In a preferred embodiment, the artificial nucleic acid, preferably the RNA, comprises the following sequence elements, preferably in a 5' to 3' orientation:

[0356] A) 5'-cap structure, preferably cap1 structure;

[0357] B) 5'-UTR, preferably selected from or derived from the 5'-UTR of the HSD17B4 gene, or a fragment thereof;

[0358] C) CDs encoding at least one antigenic peptide, said antigenic peptide being selected from peptides or proteins encoded by long non-coding RNA (lncRNA), said antigenic peptide comprising or constituting an amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, or an immunogenic fragment or variant thereof;

[0359] D) 3'-UTR, preferably selected from or derived from the 3'-UTR of the PSMB3 gene, or a fragment thereof;

[0360] E) Optionally, histone stem-loop; and

[0361] F) poly(A) sequence, preferably containing about 100 A nucleotides.

[0362] In a particularly preferred embodiment, the nucleic acid is mRNA, which contains the following sequence elements in the 5' to 3' orientation:

[0363] A) 5'-cap structure, preferably cap1 structure;

[0364] B) 5'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a fragment or variant of SEQ ID NO: 13 or any of these.

[0365] C) cds, comprising a nucleic acid sequence encoding at least one antigenic peptide selected from or derived from peptides or proteins encoded by long non-coding RNA (lncRNA), wherein the nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants;

[0366] D) 3'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 67, or a fragment or variant thereof;

[0367] E) Optionally, histone stem-loop; and

[0368] F) poly(A) sequence, preferably containing about 100 A nucleotides.

[0369] In a particularly preferred embodiment, the nucleic acid is mRNA, which contains the following sequence elements in the 5' to 3' orientation:

[0370] A) 5'-cap structure, preferably cap1 structure;

[0371] B) 5'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a fragment or variant of SEQ ID NO: 13 or any of these.

[0372] C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from or derived from peptides or proteins of neoantigens as defined herein, wherein the nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 449 to SEQ ID NO: 511, or fragments or variants thereof;

[0373] D) 3'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the fragment or variant thereof of SEQ ID NO: 67.

[0374] E) Optionally, histone stem-loop; and

[0375] F) poly(A) sequence, preferably containing about 100 A nucleotides.

[0376] 2: A pharmaceutical composition comprising at least one nucleic acid encoding a tumor antigen:

[0377] In a second aspect, the present invention provides a pharmaceutical composition comprising at least one artificial nucleic acid encoding at least one tumor antigen as defined in the first aspect.

[0378] It is worth noting that the features and implementation schemes described in the context of the first aspect must be read and understood as suitable implementation schemes of the pharmaceutical composition of the second aspect, and vice versa.

[0379] In the context of this invention, "composition" refers to any type of composition in which a specified component (e.g., a nucleic acid encoding at least one tumor antigen) may be incorporated, optionally together with any other component, typically together with at least one pharmaceutically acceptable carrier or excipient. The composition may be a dry composition, such as a powder, granules, or a solid lyophilized form. Alternatively, the composition may be in liquid form, and each component may be incorporated independently in a dissolved or dispersed form.

[0380] Preferably, at least one artificial nucleic acid in the pharmaceutical composition is selected from RNA as further defined in the first aspect. In a particularly preferred embodiment, at least one nucleic acid in the pharmaceutical composition is selected from mRNA as further defined in the first aspect.

[0381] In a preferred embodiment, the pharmaceutical composition comprises a plurality of or at least one type of nucleic acid (e.g., RNA type), preferably wherein each type of nucleic acid encodes a different tumor antigen.

[0382] Preferably, the pharmaceutical composition defined herein may comprise 2 to 10 nucleic acid types, each as defined in the first aspect, preferably 2 to 5 nucleic acid types, each as defined in the first aspect, wherein each of the 2 to 10 nucleic acid types, preferably each of the 2 to 5 nucleic acid types, encodes at least one different tumor antigen.

[0383] Suitablely, nucleic acid types encoding different tumor antigens are selected to obtain increased coverage in specific cancer diseases, such as NSCLC or melanoma. Suitablely, the coverage value of all different tumor antigens is greater than 0.5, preferably greater than 0.6, and more preferably greater than 0.7.

[0384] In this preferred embodiment within the context, different tumor antigens (provided by nucleic acid sequences) are selected to cover different HLA types. Thus, the different tumor antigens are selected from at least one tumor antigen containing at least one epitope binding to HLA-A type as defined herein, at least one tumor antigen containing at least one epitope binding to HLA-B type as defined herein, and at least one tumor antigen containing at least one epitope binding to HLA-C type as defined herein.

[0385] In various embodiments, at least one artificial nucleic acid, preferably at least one RNA of a pharmaceutical composition, is formulated together with a pharmaceutically acceptable carrier or excipient.

[0386] Formulation / Combination:

[0387] In a preferred embodiment, at least one nucleic acid, preferably at least one RNA, is complexed or bound with at least one other compound to obtain the formulated composition. In this context, the formulation may function as a transfection agent. The formulation in this context may also function to protect the nucleic acid from degradation, for example, to allow for storage, transportation, etc.

[0388] In the embodiments, at least one nucleic acid, preferably at least one RNA, is formulated with at least one compound such as peptide, protein, lipid, polysaccharide and / or polymer.

[0389] In an embodiment, at least one artificial nucleic acid, preferably at least one RNA, is formulated with at least one cationic (cationic or preferably ionizable) compound or polycationic compound (cationic or preferably ionizable). In a preferred embodiment, at least one artificial nucleic acid, preferably at least one RNA, is complexed or bound, or at least partially complexed or bound, with one or more cationic (cationic or preferably ionizable) compounds or polycationic compounds.

[0390] The term "cationic compound or polycationic compound" refers to a molecule that carries a positive charge in a pH range of about 1 to 9, about 3 to 8, about 4 to 8, about 5 to 8, more preferably about 6 to 8, even more preferably about 7 to 8, and most preferably at physiological pH, for example, in the range of about 7.2 to about 7.5. Therefore, cationic components, such as cationic peptides, cationic proteins, cationic polymers, cationic polysaccharides, and cationic lipids, can be any compound or polymer that carries a positive charge under physiological conditions. "Cationic peptides or proteins or polycationic peptides or proteins" may contain at least one positively charged amino acid, or more than one positively charged amino acid, such as those selected from Arg, His, Lys, or Orn. ​​Therefore, "polycationic" components exhibiting more than one positive charge under given conditions are also within the scope of this invention.

[0391] In a preferred embodiment, at least one cationic compound or polycationic compound is selected from cationic polymers or polycationic polymers, cationic polysaccharides or polycationic polysaccharides, cationic lipids or polycationic lipids, cationic proteins or polycationic proteins, cationic peptides or polycationic peptides, or any combination thereof.

[0392] In a particularly preferred embodiment, at least one artificial nucleic acid, preferably at least one RNA, of the pharmaceutical composition is formulated in a lipid-based carrier.

[0393] In the context of this invention, the term "lipid-based carrier" includes lipid-based delivery systems for nucleic acids (e.g., RNA) that contain lipid components. Lipid-based carriers may additionally contain other components suitable for encapsulating / incorporating / complexing nucleic acids (e.g., RNA), including cationic polymers or polycationic polymers, cationic polysaccharides or polycationic polysaccharides, cationic proteins or polycationic proteins, cationic peptides or polycationic peptides, or any combination thereof.

[0394] At least one nucleic acid, preferably RNA, may be wholly or partially incorporated into or encapsulated in a lipid-based carrier, wherein the at least one nucleic acid may be located within the interior space of the lipid-based carrier, within the lipid layer / membrane of the lipid-based carrier, or bound to the exterior of the lipid-based carrier. Incorporating a nucleic acid into a lipid-based carrier can be termed "encapsulation".

[0395] The term "encapsulated," such as incorporated, complexed, encapsulated, partially encapsulated, bound, or partially bound, refers to nucleic acids, preferably RNA, that are substantially stably bound to one or more lipids to form a lipid-based carrier (e.g., a larger complex or assembly), preferably without covalent binding of nucleic acids. The lipid-based carrier—encapsulated nucleic acid (e.g., RNA)—may be wholly or partially located within the lipid-based carrier (e.g., the lipid portion and / or internal space) and / or within the lipid layer / membrane of the lipid-based carrier. Encapsulating nucleic acids (e.g., RNA) into a lipid-based carrier is also referred to herein as "incorporation," as the nucleic acid (e.g., RNA) is preferably contained within the lipid-based carrier. Without wishing to be bound by theory, the purpose of incorporating or encapsulating nucleic acids into a lipid-based carrier may be to protect the nucleic acid from environmental influences that may contain enzymes, chemicals, or conditions that degrade the nucleic acid (e.g., RNA). Furthermore, incorporating nucleic acids into a lipid-based carrier can promote the uptake of nucleic acids and their release from endosomal compartments, thus enhancing the therapeutic effects of the nucleic acid (e.g., RNA) when applied to cells or objects.

[0396] In the embodiments, the lipid-based carrier is selected from liposomes, lipid nanoparticles, lipid complexes, solid lipid nanoparticles, lipid-polyplexes, and / or nanoliposomes. In a preferred embodiment, the lipid-based carrier is lipid nanoparticles (LNPs).

[0397] In a particularly preferred embodiment, lipid nanoparticles encapsulate at least one nucleic acid, preferably at least one RNA of the present invention.

[0398] In a preferred embodiment, the lipid-based carrier comprises one or more lipids selected from at least one lipid that reduces aggregation, at least one cationic lipid, at least one neutral lipid or phospholipid, or at least one steroid or steroid analogue, or any combination thereof.

[0399] In a preferred embodiment, the lipid-based carrier comprises (i) lipids that reduce aggregation, (ii) cationic or ionizable lipids, and (iii) neutral lipids / phospholipids or steroids / steroid analogs.

[0400] In a particularly preferred embodiment, the lipid-based carrier comprises (i) lipids that reduce aggregation, (ii) cationic or ionizable lipids, (iii) neutral lipids or phospholipids, and (iv) steroids or steroid analogs.

[0401] Reduced aggregation of lipids / polymer-conjugated lipids:

[0402] In a preferred embodiment, the lipid-based carrier comprises at least one lipid or portion that reduces aggregation.

[0403] The term "agglomeration-reducing portion" refers to a molecule containing a portion suitable for reducing or preventing the aggregation of lipid-based carriers. The term "agglomeration-reducing lipid" refers to a molecule containing both a lipid portion and a portion suitable for reducing or preventing the aggregation of lipid-based carriers. Under storage conditions or during formulation, lipid-based carriers may experience charge-induced aggregation, which can be undesirable for the stability of the lipid-based carrier. Therefore, it may be necessary to include compounds or portions capable of reducing aggregation, for example, by sterically stabilizing the lipid-based carrier. This steric stabilization occurs when a compound having a sterically large but uncharged portion shields or blocks the charged portion of the lipid-based carrier from accessing other lipid-based carriers in the composition. In the context of this invention, the stabilization of lipid-based carriers is achieved by including lipids, which may contain lipids with sterically large groups, preferably located externally to the lipid-based carrier after its formation. Suitable groups that reduce aggregation include hydrophilic groups, such as monosialotetrahexosylganglioside GM1, polyamide oligomers (PAO), or certain polymers, such as poly(oxyalkylene oxide), such as poly(ethylene glycol) or poly(propylene glycol).

[0404] Lipids containing polymers as groups that reduce aggregation are referred to herein as "polymer-conjugated lipids".

[0405] The term "polymer-conjugated lipid" refers to a molecule that comprises both a lipid moiety and a polymer moiety, wherein the polymer is adapted to reduce or prevent the aggregation of lipid-based carriers containing the RNA. A polymer should be understood as a substance or material composed of very large molecules or macromolecules, consisting of a number of repeating subunits. Suitable polymers in the context of this invention may be hydrophilic polymers. Examples of polymer-conjugated lipids are PEGylated lipids or PEG-conjugated lipids.

[0406] In a preferred embodiment, the lipid-based carrier comprises a reduced-aggregation lipid selected from polymer-conjugated lipids.

[0407] In a preferred embodiment, the polymer-conjugated lipid is a PEG-conjugated lipid (or a PEGylated lipid, PEG lipid).

[0408] The average molecular weight of the PEG moiety in the PEG-conjugated lipid preferably ranges from about 500 Daltons to about 8000 Daltons (e.g., about 1000 Daltons to about 4000 Daltons). In a preferred embodiment, the average molecular weight of the PEG moiety is about 2000 Daltons. The PEG-conjugated lipid may be selected from or derived from 1,2-dimyristoyl-rac-glycerol-3-methoxypolyethylene glycol-2000 (PEG2000 DMG or DMG-PEG 2000), C10-PEG2K, or Cer8-PEG2K. The PEG-conjugated lipid may also be selected from or derived from formula (IVa) of WO2018078053. Suitable PEG-conjugated lipids of formula (IVa) have the chemical name 2[(polyethylene glycol)-2000]-N,N-tetracosylacetamide, also known as ALC-0159.

[0409] In a preferred embodiment, the lipid-based carrier comprises lipids that reduce aggregation, wherein the lipids that reduce aggregation are not PEG-conjugated lipids. Therefore, the lipids that reduce aggregation can be suitably selected from PEG-free lipids, such as lipids without PEG polymer conjugations.

[0410] In a preferred embodiment, the lipids that reduce aggregation (or polymer-conjugated lipids) are PEG-free lipids that contain polymers different from PEG.

[0411] In the context of this invention, PEG-free lipids may be selected from or derived from POZ lipids. In a preferred embodiment, POZ lipids or, respectively, polymer-conjugated lipids are described in WO2023031394, the entire disclosure of which is incorporated herein by reference. In particular, the disclosure relating to polymer-conjugated lipids as described in any one of claims 1 to 8 of WO2023031394 is incorporated herein by reference.

[0412] In the implementation scheme, the polymer-conjugated lipid is a PEG-free lipid selected from POZ lipids.

[0413] Therefore, in the embodiments, the polymer-conjugated lipid is a "POZ lipid", which is preferably defined as a compound according to formula (POZ): [H] - [connector] - [M], wherein

[0414] [H] is a homopolymer moiety containing at least one polyoxazoline (POZ) monomer unit.

[0415]

[0416] Wherein R is a C1-9 alkyl or C2-9 alkenyl, preferably C1, and the average value of n is in the range of 2 to 200, preferably 20 to 100, more preferably 24 to 26 or 45 to 50; [connector] is an optional connector group; [M] is a lipid moiety.

[0417] In a preferred embodiment within the context of POZ lipids, the lipids that reduce aggregation are selected from or derived from PMOZ 1, PMOZ 2, PMOZ 3, PMOZ 4 or PMOZ 5 of WO2023031394.

[0418] In a preferred embodiment, the lipids that reduce aggregation are selected from or derived from PMOZ4 according to the following formula:

[0419] .

[0420] In a preferred embodiment, at least one lipid that reduces aggregation is selected from DMG-PEG 2000, C10-PEG2K, Cer8-PEG2K, or POZ lipids such as PMOZ4. In other preferred embodiments, at least one lipid that reduces aggregation is selected from ALC-0159.

[0421] Cationic lipids:

[0422] In a preferred embodiment, the lipid-based carrier comprises at least one cationic lipid or an ionizable lipid.

[0423] The cationic or ionizable lipids in lipid-based carriers can be cationic or ionizable, meaning they protonate when the pH is below the pK of the ionizable group of the lipid, but gradually become more neutral at higher pH values. At pH values ​​below the pK, the lipid can bind to negatively charged nucleic acids. In some embodiments, the cationic lipid comprises zwitterionic lipids that exhibit a positive charge when the pH decreases.

[0424] In a preferred embodiment, the lipid-based carrier comprises cationic lipids or ionizable lipids, which preferably carry a net positive charge at physiological pH, and more preferably, the cationic lipids or ionizable lipids comprise tertiary nitrogen groups or quaternary nitrogen groups. Therefore, in a preferred embodiment, the lipid-based carrier comprises cationic lipids or ionizable lipids selected from amino lipids, preferably wherein the amino lipids comprise tertiary amine groups.

[0425] In a preferred embodiment, at least one cationic lipid or ionizable lipid is selected from or derived from formula (III-1):

[0426] .

[0427] Preferably, one of L1 or L2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -SS-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, -NRaC(=O)NRa-, -OC(=O)NRa-, or -NRaC(=O)O-, and the other of L1 or L2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -SS-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, -NRaC =O)NRa-, -OC(=O)NRa- or -NRaC(=O)O- or directly bonded; G1 and G2 are each independently unsubstituted C1 to C12 alkylene or C1 to C12 alkenyl; G3 is C1 to C24 alkylene, C1 to C24 alkenyl, C3 to C8 cycloalkylene, C3 to C8 cycloalkenyl; Ra is H or C1 to C12 alkyl; R1 and R2 are each independently C6 to C24 alkyl or C6 to C24 alkenyl; R3 is H, OR5, CN, C(=O)OR4, OC(=O)R4 or -NR5C(=O)R4; R4 is C1 to C12 alkyl; R5 is H or C1 to C6 alkyl; and x is 0, 1 or 2.

[0428] In some embodiments, cationic or ionizable lipids may be selected from lipids disclosed in WO2018078053 (i.e., lipids derived from Formulas I, II, and III of WO2018078053, or lipids as specified in claims 1 to 12 of WO2018078053), the disclosure of which is incorporated herein by reference in its entirety. In this context, lipids disclosed in Table 7 of WO2018078053 (e.g., lipids derived from Formulas I-1 to I-41) and lipids disclosed in Table 8 of WO2018078053 (e.g., lipids derived from Formulas II-1 to II-36) may be suitably used in the context of this invention. Therefore, Formulas I-1 to I-41 and Formulas II-1 to II-36 of WO2018078053, and the specific disclosures associated therewith, are incorporated herein by reference.

[0429] In an embodiment, the lipid-based carrier of the pharmaceutical composition comprises a cationic lipid selected from or derived from structures III-1 to III-36 of Table 9 of WO2018078053. Therefore, formulas III-1 to III-36 of WO2018078053, and the specific disclosures therein, are incorporated herein by reference.

[0430] In the embodiments, the lipid-based carrier comprises a cationic lipid selected from or derived from Formula III-3 of WO2018078053. The preferred lipid of Formula III-3 has the chemical name ((4-hydroxybutyl)azadiyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate), also known as ALC-0315, or CAS number 2036272-55-4.

[0431] Other suitable cationic lipids may be selected from or derived from the cationic lipids of PCT claims 1 to 14 of WO2021123332 or Table 1 of WO2021123332, the disclosure of which relates to claims 1 to 14 of WO2021123332 or Table 1 is incorporated herein by reference. Therefore, suitable cationic lipids may be selected from or derived from the cationic lipids of compounds 1 to 27 (C1 to C27) of Table 1 of WO2021123332.

[0432] In one embodiment, the lipid-based carrier comprises a cationic lipid selected from or derived from (COATSOME® SS-EC) SS-33 / 4PE-15 (see C23 in Table 1 of WO2021123332). In other embodiments, the lipid-based carrier comprises a cationic lipid selected from or derived from HEXA-C5DE-PipSS (see C2 in Table 1 of WO2021123332). In a preferred embodiment, the lipid-based carrier comprises a cationic lipid selected from or derived from compound C26 disclosed in Table 1 of WO2021123332.

[0433] In other embodiments, the lipid-based carrier comprises a cationic lipid selected from or derived from 9-heptadecyl 8-{(2-hydroxyethyl)[6-oxo-6-(undecapoxy)hexyl]amino}octanoate, also known as SM-102.

[0434] Therefore, in a preferred embodiment, the lipid-based carrier, preferably the LNP, comprises a cationic lipid selected from or derived from the aforementioned compounds C26, SM-102, SS-33 / 4PE-15, or HEXA-C5DE-PipSS. In other preferred embodiments, the lipid-based carrier, preferably the LNP, comprises a cationic lipid selected from or derived from ALC-0315.

[0435] Neutral lipids:

[0436] In a preferred embodiment, the lipid-based carrier (e.g., LNP) contains at least one neutral lipid or phospholipid.

[0437] The term "neutral lipid" refers to any of a variety of lipids that exist as uncharged or neutral zwitterions at physiological pH. Neutral lipids may be selected from DHPC, DOPC, DPPC, DOPG, DPPG, DOPE, POPC, POPE, DOPE-mal, DPPE, DMPE, DSPE, 16-O-monomethylPE, 16-O-dimethylPE, 18-1-transPE, SOPE, transDOPE, 1,2-diphydanyl-sn-glycerol-3-phosphate ethanolamine (DPhyPE), DPhyPS (1,2-diphydanyl-sn-glycerol-3-phosphate-L-serine), or mixtures thereof.

[0438] In a preferred embodiment, the lipid-based carrier comprises a neutral lipid selected from or derived from DSPC, DHPC, DPhyPE, or DPhyPS. DSPC is preferred in this context.

[0439] Steroids, steroid analogs or sterols:

[0440] In a preferred embodiment, the lipid-based carrier comprises a steroid, a steroid analogue, or a steroid.

[0441] In the embodiments, the steroid, steroid analogue, or steroid is derived from or selected from cholesterol, cholesterol hemisuccinate (CHEMS), or any derivative thereof. In a preferred embodiment, the lipid-based carrier comprises cholesterol.

[0442] Lipid-based carrier compositions:

[0443] In the implementation scheme, a lipid-based carrier comprising at least one nucleic acid, preferably at least one RNA, and preferably an LNP, comprising...

[0444] (i) at least one cationic lipid or ionizable lipid, preferably as defined herein;

[0445] (ii) at least one or two (e.g., two different) neutral lipids or phospholipids, preferably as defined herein;

[0446] (iii) at least one steroid or steroid analogue, preferably as defined herein; and

[0447] (iv) At least one lipid that reduces aggregation, preferably as defined herein.

[0448] In the implementation scheme, a lipid-based carrier comprising at least one nucleic acid, preferably at least one RNA, and preferably an LNP, comprising...

[0449] (i) at least one cationic lipid selected from or derived from C26, SS-33 / 4PE-15, HEXA-C5DE-PipSS, or SM-102;

[0450] (ii) At least one or two (e.g., two different) neutral lipids selected from or derived from DSPC, DHPC, DPhyPE, DPhyPS;

[0451] (iii) at least one steroid or steroid analogue selected from or derived from cholesterol; and

[0452] (iv) At least one lipid selected from or derived from DMG-PEG 2000, C10-PEG2K, Cer8-PEG2K, or ALC-0159 or “PMOZ 4” to reduce aggregation; and wherein the lipid-based carrier encapsulates the nucleic acid (e.g., the RNA).

[0453] In the implementation scheme, a lipid-based carrier comprising at least one nucleic acid, preferably at least one RNA, and preferably the LNP, comprising...

[0454] (i) at least one cationic lipid selected from or derived from ALC-0315;

[0455] (ii) At least one neutral lipid selected from or derived from DSPC;

[0456] (iii) at least one steroid or steroid analogue selected from or derived from cholesterol; and

[0457] (iv) At least one lipid selected from or derived from ALC-0159 that reduces aggregation; and wherein the lipid-based carrier encapsulates the nucleic acid (e.g., the RNA).

[0458] In the implementation scheme, a lipid-based carrier comprising at least one nucleic acid, preferably at least one RNA, contains...

[0459] (i) Cationic lipids selected from C26;

[0460] (ii) Neutral lipids selected from DPhyPE and neutral lipids selected from DPhyPS;

[0461] (iii) Steroids or steroid analogues selected from cholesterol; and

[0462] (iv) A lipid selected from POZ lipids as defined herein, preferably PMOZ 4; and wherein the lipid-based carrier encapsulates the nucleic acid (e.g., the RNA).

[0463] In the implementation scheme, cationic lipids (as defined herein), neutral lipids (as defined herein), steroids or steroid analogs (as defined herein), and / or lipids that reduce aggregation (as defined herein) may be combined in various relative proportions.

[0464] In an embodiment, the lipid-based carrier, preferably the LNP, comprises (i) to (iv) a molar ratio of about 20% to 60% cationic lipids or ionizable lipids, about 5% to 25% neutral lipids, about 25% to 55% steroids or steroid analogs, and about 0.5% to 15% lipids that reduce aggregation, such as polymer-conjugated lipids, wherein the lipid-based carrier encapsulates the at least one nucleic acid, preferably at least one RNA.

[0465] In a preferred embodiment, the lipid-based carrier, preferably an LNP, comprises (i) to (iv) a molar ratio of about 45% to 55% cationic lipids or ionizable lipids, about 5% to 15% neutral lipids, about 35% to 45% steroids or steroid analogs, and about 0.5% to 2.5% lipids that reduce aggregation, such as polymer-conjugated lipids, wherein the lipid-based carrier preferably encapsulates the at least one nucleic acid, preferably at least one RNA.

[0466] In a more preferred embodiment, the lipid-based carrier, preferably the LNP, comprises (i) to (iv) a molar ratio of about 47% to 51% cationic lipids or ionizable lipids, about 8% to 12% neutral lipids, about 38% to 42% steroids or steroid analogs, and about 0.75% to 1.75% lipids that reduce aggregation, such as polymer-conjugated lipids, wherein the lipid-based carrier preferably encapsulates the at least one nucleic acid, preferably at least one RNA.

[0467] In the implementation scheme, the lipid-based carrier preferably comprises at least one nucleic acid, preferably at least one RNA, in an LNP, which contains...

[0468] (i) at least one cationic lipid selected from ALC-0315;

[0469] (ii) At least one neutral lipid selected from DSPC;

[0470] (iii) at least one steroid or steroid analogue selected from cholesterol; and

[0471] (iv) At least one aggregation-reducing lipid selected from ALC-0159; and wherein the lipid-based carrier encapsulates the nucleic acid (e.g., the RNA), preferably wherein the molar ratio of i) to (iv) is about 47.4% cationic lipid, about 10% neutral lipid, about 40.9% steroid or steroid analogue, and about 1.7% aggregation-reducing lipid, preferably wherein the lipid-based carrier encapsulates the nucleic acid (e.g., the RNA).

[0472] In a preferred embodiment, the lipid-based carrier, preferably containing at least one nucleic acid molecule, preferably RNA, comprises...

[0473] (i) Cationic lipid C26 (VitE-C4DE-Pip-thioether);

[0474] (ii) Neutral lipids DPhyPE and DPhyPS;

[0475] (iii) Steroids or steroid analogues (cholesterol); and

[0476] (iv) The aggregation-reducing lipid PMOZ4, preferably wherein the molar ratio of i) to (iv) is about 49% cationic lipid, about 10% neutral lipid, about 40% steroid or steroid analog, and about 1% aggregation-reducing lipid, preferably wherein the lipid-based carrier encapsulates the at least one nucleic acid molecule, preferably the RNA.

[0477] The amount of lipids contained in a lipid-based carrier can be selected taking into account the amount of nucleic acid cargo. In one embodiment, these amounts are selected such that the N / P ratio of the lipid-based carrier encapsulating the nucleic acid ranges from about 0.1 to about 50. The N / P ratio is defined as the molar ratio of nitrogen atoms (“N”) of the basic nitrogen-containing group of the lipid to phosphate groups (“P”) of the nucleic acid used as cargo. The N / P ratio can be calculated based on the following: for example, assuming a statistical distribution of nucleic acid bases, 1 µg of nucleic acid typically contains about 3 nmol of phosphate residues. The “N” value of the lipid or lipid-like compound can be calculated based on its molecular weight and the relative amounts of permanent cationic groups and (if present) cationic groups.

[0478] In some embodiments, the N / P ratio can range from about 1 to about 50. In a preferred embodiment, this range is from about 5 to about 20. In some embodiments, the N / P ratio is about 17. In some embodiments, the N / P ratio is about 14. In some embodiments, the N / P ratio is about 6.

[0479] In various embodiments, the pharmaceutical composition comprises a lipid-based carrier (encapsulating nucleic acid, preferably RNA as defined herein) having a defined size (particle size, uniform particle size distribution).

[0480] The particle size of lipid-based carriers is typically described herein as the Z-mean particle size. The term "Z-mean particle size" refers to the average diameter of the particles measured by dynamic light scattering (DLS), using a so-called cumulant algorithm for data analysis, which provides the so-called Z-mean with length and a dimensionless polydispersity index (PDI) as a result. The terms "dynamic light scattering" or "DLS" refer to a method for analyzing particles in liquids, where the liquid is typically illuminated by a monochromatic light source, and where light scattered by particles in the liquid is detected. DLS schemes and instruments are known in the art.

[0481] In a preferred embodiment, the lipid-based carrier, preferably the LNP, has a Z-average particle size of less than 400 nm, more preferably less than 300 nm, and even more preferably less than 200 nm.

[0482] In the embodiments, the lipid-based carrier, preferably LNP, has a Z-average particle size ranging from about 50 nm to about 200 nm, preferably from about 50 nm to about 150 nm, and more preferably from about 50 nm to about 120 nm.

[0483] Preferably, the composition contains less than about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% of a lipid-based carrier having a particle size greater than about 500 nm. Preferably, the composition contains less than about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% of LNP having a particle size less than about 20 nm.

[0484] In the embodiments, the lipid-based carrier exhibits a zeta potential in the range of -50 mV to +50 mV, preferably -25 mV to +25 mV, more preferably -10 mV to +10 mV, and most preferably -5 mV to +5 mV.

[0485] In the embodiments, the lipid-based carrier, preferably LNP, has a polydispersity index (PDI) in the range of 0.1 to 0.5. In a preferred embodiment, the PDI value is less than about 0.3, preferably less than about 0.2. Typically, the PDI is determined by dynamic light scattering.

[0486] In the implementation scheme, at least 70%, 80%, 90%, and 95% of the nucleic acid (e.g., RNA) is encapsulated in a lipid-based carrier such as LNP. The encapsulation percentage can be determined using the RiboGreen assay known in the art.

[0487] In the embodiments, multiple lipid-based carriers have a layered morphology and / or a bilayer morphology. In the embodiments, at least 80%, 85%, 90%, and 95% of the lipid-based carriers have a spherical morphology.

[0488] In a preferred embodiment, the lipid-based carrier, preferably the LNP, has an uncharged surface at pH 7.

[0489] Administration of pharmaceutical compositions or nucleic acids

[0490] Appropriately, when a pharmaceutical composition or nucleic acid is applied to a cell, tissue, or object, at least one tumor antigen is produced in an amount sufficient to induce an antigen-specific immune response in said cell, tissue, or object.

[0491] In a preferred embodiment, the administration is intramuscular, intratumoral, or intravenous, preferably intramuscular, intratumoral, or intravenous injection.

[0492] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or objects, an encoded tumor antigen is generated, preferably in an amount sufficient to induce an antigen-specific immune response in the cells, tissues, or objects.

[0493] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, it generates encoded tumor antigens and induces increased immunogenicity in the subject.

[0494] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, it generates encoded tumor antigens and induces epitope-specific or antigen-specific CD8+ T cells in the subject.

[0495] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, it generates encoded tumor antigens and induces epitope-specific or antigen-specific CD4+ T cells in the subject.

[0496] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, it generates encoded tumor antigens and induces humoral immunity, such as antibody titers against tumor antigens encoded in the subject and / or increased antibody diversity.

[0497] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, an encoded tumor antigen is generated, and IFN-γ production by CD8+ T cells is increased when the subject is exposed to the encoded tumor antigen.

[0498] In a preferred embodiment, when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, it generates encoded tumor antigens and increases the presentation of encoded tumor antigens on MHC molecules in the subject.

[0499] The term "presentation" of encoded polypeptides or peptides refers to a higher number and / or variety of immunogenic peptides and / or stable peptides presented via MHC class I and MHC class II molecules following protein degradation via the proteasome mechanism, and thus involves cellular immunity, such as T cell activation. Presentation of encoded polypeptides or peptides on MHC class I and MHC class II molecules is increased on cells, including immune cells (e.g., T cells), antigen-presenting cells (e.g., dendritic cells, macrophages, engineered antigen-presenting cells), MHC class I expressing cells, MHC class II expressing cells, or any combination thereof.

[0500] For example, administration of a pharmaceutical composition or nucleic acid results in an increased immunogenicity presentation of the encoded tumor antigen on the MHC molecule, with an increase of about 0.1% to about 100% (e.g., about 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or about 100%) compared to a subject that has not been administered nucleic acid / composition.

[0501] In some embodiments, administration of the pharmaceutical composition or nucleic acid results in an increase of approximately 2 to approximately 100 times (e.g., approximately 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 25, 30, 40, 50, 60, 70, 80, 90, or approximately 100 times) in the immunogenicity of the encoded tumor antigen or in the presentation of the MHC molecule compared to a subject not administered the nucleic acid / composition.

[0502] 3: Combinations of tumor antigens / tumor antigens:

[0503] In a third aspect, the present invention provides a peptide tumor antigen or a composition comprising at least one peptide tumor antigen.

[0504] It is worth noting that the features and embodiments described in the context of the first aspect (nucleic acids of the present invention) or the second aspect (pharmaceutical compositions of the present invention) must be read and understood as suitable embodiments of the tumor antigens or compositions of the present aspect.

[0505] In a preferred embodiment, the tumor antigen comprises at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA), and / or at least one antigenic peptide selected from peptides or proteins of tumor neoantigens.

[0506] In a preferred embodiment within this context, at least one antigenic peptide is encoded by a long non-coding RNA (lncRNA), particularly lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25, or any of these immunogenic fragments or variants.

[0507] Preferred peptide tumor antigens are provided in Tables 1A and 1B (columns B or C).

[0508] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by long non-coding RNA (lncRNA), and comprises or constitutes at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these.

[0509] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of tumor neoantigens, particularly from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, PIK3CA neoantigen, NFE2L2 neoantigen, or CNOT9 neoantigen, or any immunogenic fragments or variants of these.

[0510] Preferred peptide tumor antigens are provided in Table 2 (Column C), particularly in rows 1 to 11 of Column C.

[0511] In the embodiments, at least one antigenic peptide selected from tumor neoantigens comprises or constitutes at least one amino acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, or an immunogenic fragment or immunogenic variant thereof.

[0512] In a preferred embodiment, at least one antigenic peptide selected from tumor neoantigens comprises or constitutes at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, or any immunogenic fragment or immunogenic variant thereof.

[0513] This document also provides compositions comprising at least one peptide tumor antigen as defined herein.

[0514] "A composition comprising at least one tumor antigen" refers to any type of composition in which a specified peptide tumor antigen may be incorporated, optionally together with any other component, typically together with at least one pharmaceutically acceptable carrier or excipient. The composition may be a dry composition, such as a powder, granules, or a solid lyophilized form. Alternatively, the composition may be in liquid form, and each component may be incorporated independently in a dissolved or dispersed (e.g., suspension or emulsion) form.

[0515] In various embodiments, the at least one tumor antigen of the composition is formulated together with a pharmaceutically acceptable carrier or excipient.

[0516] In embodiments, the composition comprises multiple or at least one different peptide tumor antigen as defined herein. Preferably, the composition as defined herein may comprise 2 to 10 different peptide tumor antigens as defined herein, more preferably 2 to 5 different peptide tumor antigens as defined herein.

[0517] 4: Antibodies, T cells, or TCR:

[0518] In a fourth aspect, the present invention provides antibodies, T cells, or T cell receptors (TCRs) that have been generated or stimulated with any tumor antigen or nucleic acid of any of the foregoing aspects.

[0519] It is worth noting that the features and embodiments described in the context of the first aspect (nucleic acid of the present invention), the second aspect (pharmaceutical composition of the present invention) or the third aspect (tumor antigen) must be read and must be understood as suitable embodiments of the tumor antigen or nucleic acid of the present aspect.

[0520] In the embodiments, the antibody is a therapeutic antibody generated against any tumor antigen of the present invention. Preferably, the antibody is generated against any peptide tumor antigen provided in Tables 1A and 1B (columns B or C) or any peptide tumor antigen provided in Table 2 (column C).

[0521] In the implementation scheme, the T cells are therapeutic T cells that have been stimulated with any tumor antigen or any nucleic acid as described above. Preferably, the T cells have been stimulated with any tumor antigen or any nucleic acid provided in Table 1A, Table 1B, or Table 2.

[0522] In the embodiments, the TCR is generated against any tumor antigen of the present invention (e.g., an antigenic peptide loaded on the MHC). Preferably, the TCR is generated against any peptide tumor antigen provided in Table 1A or Table 1B (column B or column C) or any peptide tumor antigen provided in Table 2 (column C).

[0523] 5: Combinations:

[0524] In a fifth aspect, the present invention provides a combination comprising at least two or more of the treatment methods described above.

[0525] It is worth noting that the features and embodiments described in the context of the first aspect (nucleic acids of the present invention), the second aspect (pharmaceutical compositions of the present invention), the third aspect (tumor antigens), or the fourth aspect (antibodies, T cells, TCRs) must be read and understood as suitable embodiments of the combination of the fifth aspect.

[0526] In a preferred embodiment, the combination comprises at least two treatment modalities selected from the following.

[0527] a) at least one or more artificial nucleic acids as defined in the first aspect; and / or

[0528] b) at least one or more pharmaceutical compositions as defined in the second aspect; and / or

[0529] c) at least one or more tumor antigens or compositions as defined in the third aspect; and / or

[0530] d) At least one or more antibodies, T cells or TCRs as defined in the fourth aspect;

[0531] In embodiments, the combination of the present invention comprises at least one or more treatment methods as defined herein, preferably two, three, four, five, six, seven, eight or even more than eight treatment methods as defined herein.

[0532] Treatments can be applied spatially and / or temporally in a staggered manner.

[0533] 6: Reagent kits or complete reagent kits:

[0534] In a sixth aspect, the present invention provides a reagent kit or a complete kit.

[0535] It is worth noting that the features and embodiments described in the context of the first aspect (nucleic acids of the present invention), the second aspect (pharmaceutical compositions of the present invention), the third aspect (tumor antigens), the fourth aspect (antibodies, T cells, TCRs), or the fifth aspect (combinations) must be read and understood as suitable embodiments of the kits or kits of the sixth aspect.

[0536] In a preferred embodiment, the kit or kit includes

[0537] i) at least one or more artificial nucleic acids as defined in the first aspect; and / or

[0538] ii) at least one or more pharmaceutical compositions as defined in the second aspect; and / or

[0539] iii) at least one or more tumor antigens or compositions as defined in the third aspect; and / or

[0540] iv) at least one or more antibodies, T cells or TCRs as defined in the fourth aspect;

[0541] In the implementation scheme, the kit or kit package includes a liquid carrier for dissolution.

[0542] In the implementation scheme, the kit or kit package includes technical instructions providing information on component administration and dosage. The technical instructions for the kit may include information on administration, dosage, and patient population. Such kits, preferably kit packages, can be used for any application or use, such as those mentioned herein, preferably with therapeutic agents i) to iv) for the treatment of cancer or cancer-related diseases, disorders, or conditions.

[0543] 7. Medical uses:

[0544] In another aspect, the present invention relates to the medical use of the therapeutic agents described above.

[0545] It is worth noting that the features and embodiments described in the context of the kits or kits of the first aspect (nucleic acids of the present invention), the second aspect (pharmaceutical compositions of the present invention), the third aspect (tumor antigens), the fourth aspect (antibodies, T cells, TCRs), the fifth aspect (combinations), or the sixth aspect must be read and understood as suitable embodiments for any medical use presented in the following aspects.

[0546] Therefore, the present invention provides artificial nucleic acids in the first aspect, pharmaceutical compositions in the second aspect, tumor antigens or compositions of tumor antigens in the third aspect, antibodies, T cells or TCRs in the fourth aspect, combinations in the fifth aspect, or kits or kits in the sixth aspect, which are used as medicines, for example for treating or preventing diseases, disorders or conditions of a subject.

[0547] In a preferred embodiment, the use can be for human medical purposes and for veterinary medical purposes, with human medical purposes being preferred.

[0548] In a preferred embodiment, the use may be for human medical purposes, particularly for infants, newborns, immunocompromised recipients, pregnant and lactating women, and the elderly.

[0549] In a preferred embodiment, the use is for human medical purposes for a person suffering from cancer.

[0550] Medical uses for treating or preventing cancer

[0551] In another aspect, the present invention provides artificial nucleic acids of the first aspect, pharmaceutical compositions of the second aspect, tumor antigens or compositions of tumor antigens of the third aspect, antibodies, T cells or TCRs of the fourth aspect, combinations of the fifth aspect, or kits or kits of the sixth aspect, which are used as medicines for treating or preventing cancer or any cancer-related disease, disorder or ailment in a subject.

[0552] It is worth noting that the features and implementation schemes described in any of the foregoing contexts can be read and must be understood as appropriate implementation schemes of the present aspects.

[0553] As used herein, the term "cancer" refers to a tumor characterized by uncontrolled and often rapid proliferation of cells that tend to invade surrounding tissues and metastasize to distant sites of the body. The term includes both benign and malignant tumors. Malignancy in cancer is typically characterized by anaplasia, invasiveness, and metastasis; while benign and malignant tumors typically do not possess these characteristics. The term includes tumors characterized by tumor growth as well as cancers of the blood and lymphatic systems. According to this disclosure, the term "cancer" also includes cancer metastasis, such as cancer metastasis associated with HNSCC, NSCLC, or melanoma.

[0554] In some implementations, cancer treatment or prevention in a subject is personalized. Therefore, before treatment with the appropriate therapeutic agent, the patient's cancer is analyzed, for example, by sequencing.

[0555] In various implementation schemes, nucleic acids, pharmaceutical compositions, tumor antigens or tumor antigen compositions, antibodies, T cells or TCRs, combinations, kits or kits are administered intramuscularly, intratumorally or intravenously, particularly by injection.

[0556] Medical uses for the treatment or prevention of squamous cell carcinoma

[0557] In another aspect, the present invention provides artificial nucleic acids of the first aspect, pharmaceutical compositions of the second aspect, tumor antigens or compositions of tumor antigens of the third aspect, antibodies, T cells or TCRs of the fourth aspect, combinations of the fifth aspect, or kits or kits of the sixth aspect, which are used as medicines for treating or preventing squamous cell carcinoma.

[0558] It is worth noting that the features and implementation schemes described in any of the foregoing contexts can be read and must be understood as appropriate implementation schemes of the present aspects.

[0559] In a preferred embodiment, the squamous cell carcinoma is selected from lung squamous cell carcinoma (LUSC) / squamous NSCLC (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC).

[0560] In some implementations, the treatment or prevention of squamous cell carcinoma in a subject is personalized. Therefore, before treatment with the appropriate therapeutic agent, the patient's squamous cell carcinoma is analyzed, for example, by sequencing.

[0561] Medical uses for the treatment or prevention of NSCLC

[0562] In another aspect, the present invention provides artificial nucleic acids of the first aspect, pharmaceutical compositions of the second aspect, tumor antigens or compositions of tumor antigens of the third aspect, antibodies, T cells or TCRs of the fourth aspect, combinations of the fifth aspect, or kits or kits of the sixth aspect, which are used as medicines for treating or preventing non-small cell lung cancer (NSCLC) or any disease, disorder or condition associated with NSCLC in subjects.

[0563] In the implementation plan, the use relates to the treatment of a subgroup of NSCLC, including lung adenocarcinoma (LUAD) and / or lung squamous cell carcinoma (LUSC). Preferably, the use relates to the treatment of squamous cell carcinoma (LUSC) / sqNSCLC.

[0564] It is worth noting that the features and implementation schemes described in any of the foregoing contexts can be read and must be understood as appropriate implementation schemes of the present aspects.

[0565] In the preferred embodiment within this context, at least one tumor antigen provided by artificial nucleic acid, pharmaceutical composition, tumor antigen or composition of tumor antigens, or kit or kit kit contains

[0566] a) Select at least one antigenic peptide of a peptide or protein encoded by lnc-WDR72-2:4, lnc-TRPC5-3:1, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, or any of these immunogenic fragments or variants; and / or

[0567] b) At least one antigenic peptide selected from a tumor neoantigen, which is selected from BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, PIK3CA neoantigen or NFE2L2 neoantigen, or any immunogenic fragment or variant of these, preferably selected from EGFR neoantigen or TP53 neoantigen, or any immunogenic fragment or variant of these.

[0568] In a particularly preferred embodiment within this context, at least one tumor antigen provided by an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or a kit or kit kit comprises at least one antigenic peptide encoded by a peptide or protein selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, or any of these immunogenic fragments or variants.

[0569] In various embodiments within this context, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, kits or kits are administered intramuscularly, intratumorally or intravenously, particularly by intramuscular injection.

[0570] In some implementations, the treatment or prevention of NSCLC in a subject is personalized. Therefore, before treatment with the appropriate therapeutic agent, the patient's NSCLC cancer is analyzed, for example, by sequencing.

[0571] Medical uses for the treatment or prevention of HNSCC

[0572] In another aspect, the present invention provides artificial nucleic acids of the first aspect, pharmaceutical compositions of the second aspect, tumor antigens or compositions of tumor antigens of the third aspect, antibodies, T cells or TCRs of the fourth aspect, combinations of the fifth aspect, or kits or kits of the sixth aspect, which are used as medicines for treating or preventing head and neck squamous cell carcinoma (HNSCC) or any disease, disorder or condition associated with HNSCC in subjects.

[0573] It is worth noting that the features and implementation schemes described in any of the foregoing contexts can be read and must be understood as appropriate implementation schemes of the present aspects.

[0574] In a particularly preferred embodiment within this context, at least one tumor antigen provided by an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or a kit or kit kit comprises at least one antigenic peptide encoded by a peptide or protein selected from lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, or any of these immunogenic fragments or variants.

[0575] In various embodiments within this context, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, kits or kits are administered intramuscularly, intratumorally or intravenously, particularly by intramuscular injection.

[0576] In some implementations, the treatment or prevention of HNSCC in a subject is personalized. Therefore, before treatment with the appropriate therapeutic agent, the patient's HNSCC cancer is analyzed, for example, by sequencing.

[0577] Medical uses for the treatment or prevention of melanoma

[0578] In another aspect, the present invention provides artificial nucleic acids of the first aspect, pharmaceutical compositions of the second aspect, tumor antigens or compositions of tumor antigens of the third aspect, antibodies, T cells or TCRs of the fourth aspect, combinations of the fifth aspect, or kits or kits of the sixth aspect, which are used as medicines for treating or preventing melanoma or any disease, disorder or symptom associated with melanoma in subjects.

[0579] In the implementation plan, the use relates to the treatment of cutaneous melanoma (SKCM) or uveal melanoma.

[0580] In the implementation plan, the use relates to the treatment of the BRAF subgroup and / or NRAS subgroup of melanoma.

[0581] It is worth noting that the features and implementation schemes described in any of the foregoing contexts can be read and must be understood as appropriate implementation schemes of the present aspects.

[0582] In the preferred embodiment within this context, at least one tumor antigen provided by artificial nucleic acid, pharmaceutical composition, tumor antigen or composition of tumor antigens, or kit or kit kit contains

[0583] a) Select at least one antigenic peptide of a peptide or protein encoded by lnc-TRPC5-3:1 or lnc-CLEC2D-9:1, or any of these immunogenic fragments or variants; and / or

[0584] (b) At least one antigenic peptide selected from a peptide or protein of a tumor neoantigen, wherein the tumor neoantigen is selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, CNOT9 neoantigen, preferably selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, or BRAF neoantigen, or any immunogenic fragment or variant of these.

[0585] In a particularly preferred embodiment within this context, at least one tumor antigen provided by an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or a kit or kit kit comprises at least one antigenic peptide of a peptide or protein encoded by lnc-TRPC5-3:1 or lnc-CLEC2D-9:1, or any of these immunogenic fragments or variants.

[0586] In various embodiments within this context, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, kits or kits are administered intramuscularly, intratumorally or intravenously, particularly by intramuscular injection.

[0587] In some implementations, the treatment or prevention of melanoma in a subject is personalized. Therefore, before treatment with the appropriate therapeutic agent, the patient's melanoma is analyzed, for example, by sequencing.

[0588] Medical use for the treatment or prevention of other cancer indications

[0589] In other respects, the present invention relates to the medical use of the therapeutic agents described above in the treatment or prevention of other cancer indications.

[0590] It is worth noting that the features and embodiments described in the context of the kits or kits of the first aspect (nucleic acids of the present invention), the second aspect (pharmaceutical compositions of the present invention), the third aspect (tumor antigens), the fourth aspect (antibodies, T cells, TCRs), the fifth aspect (combinations), or the sixth aspect must be read and understood as suitable embodiments for any medical use presented in the following aspects.

[0591] In another aspect, the present invention provides an artificial nucleic acid of the first aspect, a pharmaceutical composition of the second aspect, a tumor antigen or a composition of tumor antigens of the third aspect, an antibody, T cell or TCR of the fourth aspect, a combination of the fifth aspect, or a kit or kit of the sixth aspect, which are used as medicaments for treating or preventing adrenocortical carcinoma, urothelial carcinoma of the bladder, invasive breast cancer, cervical cancer (cervical squamous cell carcinoma and cervical endometrial adenocarcinoma), colonic adenocarcinoma, renal papillary cell carcinoma, lung adenocarcinoma, lymphoid tumor diffuse large B-cell lymphoma, rectal adenocarcinoma, sarcoma (especially synovial sarcoma), melanoma of the skin, thymoma, thyroid cancer, hepatocellular carcinoma, uterine carcinosarcoma or endometrial cancer, or any related disease, disorder or condition, wherein the at least one tumor antigen provided by said artificial nucleic acid, pharmaceutical composition, tumor antigen or a composition, combination, or kit of tumor antigens comprises at least one antigenic peptide selected from a peptide or protein encoded by ZC3H8-6:1, or an immunogenic fragment or variant thereof.

[0592] In another aspect, the present invention provides an artificial nucleic acid of the first aspect, a pharmaceutical composition of the second aspect, a tumor antigen or a tumor antigen composition of the third aspect, an antibody, T cell or TCR of the fourth aspect, a combination of the fifth aspect, or a kit or kit of the sixth aspect, which are used as medicaments for treating or preventing bladder urothelial carcinoma, cervical cancer (cervical squamous cell carcinoma and cervical endometrial adenocarcinoma), thymoma or endometrial cancer of the uterus, or any related disease, disorder or condition, wherein the at least one tumor antigen provided by said artificial nucleic acid, pharmaceutical composition, tumor antigen or a composition, combination, or kit of said tumor antigen comprises at least one antigenic peptide selected from a peptide or protein encoded by KCNMB2-AS1:4, or an immunogenic fragment or variant thereof.

[0593] In another aspect, the present invention provides an artificial nucleic acid of the first aspect, a pharmaceutical composition of the second aspect, a tumor antigen or a composition of tumor antigens of the third aspect, an antibody, T cell or TCR of the fourth aspect, a combination of the fifth aspect, or a kit or kit of the sixth aspect, which are used as a medicament for treating or preventing ovarian serous cystadenocarcinoma, testicular germ cell tumor, thymoma or uterine carcinosarcoma, or any related disease, disorder or condition in a subject, wherein at least one tumor antigen provided by said artificial nucleic acid, pharmaceutical composition, tumor antigen or a composition, combination of tumor antigens, or kit or kit comprises at least one antigenic peptide selected from lnc-NTF3-5:5 encoded by a peptide or protein, or an immunogenic fragment or variant thereof.

[0594] In another aspect, the present invention provides an artificial nucleic acid of the first aspect, a pharmaceutical composition of the second aspect, a tumor antigen or a composition of tumor antigens of the third aspect, an antibody, T cell or TCR of the fourth aspect, a combination of the fifth aspect, or a kit or kit of the sixth aspect, which are used as medicaments for treating or preventing bladder urothelial carcinoma, colon adenocarcinoma, esophageal cancer or gastric adenocarcinoma, or any related disease, disorder or condition in subjects, wherein at least one tumor antigen provided by said artificial nucleic acid, pharmaceutical composition, tumor antigen or a composition, combination of tumor antigens, or kit or kit comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-WDR72-2:4, or an immunogenic fragment or variant thereof.

[0595] In another aspect, the present invention provides an artificial nucleic acid of the first aspect, a pharmaceutical composition of the second aspect, a tumor antigen or a composition of tumor antigens of the third aspect, an antibody, T cell or TCR of the fourth aspect, a combination of the fifth aspect, or a kit or kit of the sixth aspect, which are used as medicaments for treating or preventing cutaneous melanoma, testicular germ cell tumors or uveal melanoma, or any related disease, disorder or condition in subjects, wherein at least one tumor antigen provided by said artificial nucleic acid, pharmaceutical composition, tumor antigen or a composition, combination of tumor antigens, or kit or kit comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-CLEC2D-9:1, or an immunogenic fragment or variant thereof.

[0596] 8. Treatment methods:

[0597] In another aspect, the present invention relates to methods for treating or preventing diseases, disorders, or ailments.

[0598] It is worth noting that embodiments relating to the preceding aspects can also be read and understood as suitable embodiments of the treatment methods of the present invention. In particular, the specific features and embodiments of the treatment methods provided herein can also be applied to the medical uses of the present invention, and vice versa.

[0599] Prevention (suppression) or treatment of disease involves suppressing the full development of a disease or condition, for example, in subjects at risk of developing a disease (such as an infection or cancer). "Treatment" refers to a therapeutic intervention that improves the signs or symptoms of a disease or pathological condition after it has begun to develop. The term "improvement," in relation to a disease or pathological condition, refers to any observable beneficial effect of treatment. Suppressing a disease can include preventing or reducing the risk of developing the disease. Beneficial effects can be demonstrated, for example, by delayed onset of clinical symptoms of the disease in susceptible subjects, a reduction in the severity of some or all of the clinical symptoms of the disease, a slowing of disease progression, an improvement in the overall health or well-being of the subject, or other parameters specific to that particular disease. "Preventative" treatment is administered to subjects who do not show signs of disease or only show early signs to reduce the risk of developing a pathological condition.

[0600] In a preferred embodiment, the present invention relates to a method for treating or preventing a disease, disorder, or ailment, wherein the method comprises administering to a subject in need an effective amount of a first aspect artificial nucleic acid, a second aspect pharmaceutical composition, a third aspect tumor antigen or a composition of tumor antigens, a fourth aspect antibody, T cells or TCRs, a fifth aspect combination, or a sixth aspect kit or kit.

[0601] As used herein, “effective” when referring to the amount of a therapeutic compound means that the amount of the compound is sufficient to produce the desired therapeutic response without undue adverse side effects (such as toxicity, irritation, or allergic reactions) and is commensurate with the benefit / risk ratio when used in the manner disclosed.

[0602] In a preferred embodiment, the disease, disorder, or condition is cancer, such as any cancer-related disease, disorder, or condition as defined herein. In a preferred embodiment, the cancer is NSCLC. In a preferred embodiment, the cancer is squamous cell carcinoma, preferably selected from squamous cell carcinoma of the lung (LUSC) / squamous non-small cell lung cancer (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC). In other preferred embodiments, the cancer is melanoma.

[0603] Preferred embodiments are provided in this context, particularly in the sections on “Medical Use for the Treatment or Prevention of Squamous Cell Carcinoma,” “Medical Use for the Treatment or Prevention of NSCLC,” “Medical Use for the Treatment or Prevention of HNSCC,” “Medical Use for the Treatment or Prevention of Melanoma,” or “Medical Use for the Treatment or Prevention of Other Cancer Indications,” which are outlined in the context of “Medical Use.”

[0604] In various implementation schemes, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, kits or kits are administered intramuscularly, intratumorally or intravenously, particularly by injection.

[0605] In some implementations, cancer treatment or prevention in a subject is personalized. Therefore, before treatment with the appropriate therapeutic agent, the patient's cancer is analyzed, for example, by sequencing.

[0606] Terms and Conditions

[0607] Preferred embodiments of the present invention are provided in the following list of numbered clauses (clauses 1 to 116):

[0608] 1. An artificial nucleic acid comprising at least one coding sequence of at least one tumor antigen, said tumor antigen comprising or constituting as

[0609] a) Select at least one antigenic peptide encoded by a long non-coding RNA (lncRNA), a peptide or protein thereof, or an immunogenic fragment or variant thereof; and / or

[0610] b) At least one antigenic peptide selected from a tumor neoantigen peptide or protein, or an immunogenic fragment or variant thereof.

[0611] 2. The artificial nucleic acid according to Clause 1, wherein the at least one antigenic peptide is selected from peptides or proteins encoded by long non-coding RNA (lncRNA), or any immunogenic fragments or variants thereof, wherein the long non-coding RNA (lncRNA) is selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, lnc-CLEC2D-9:1, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-NTF3-5:5, or LINC00893:25.

[0612] 3. The artificial nucleic acid according to claims 1 to 2, wherein the at least one antigenic peptide encoded by long non-coding RNA (lncRNA) comprises at least one T-cell epitope, preferably at least one CD8+ T-cell epitope.

[0613] 4. The artificial nucleic acid according to claims 1 to 3, wherein the at least one antigenic peptide selected from a long non-coding RNA (lncRNA) encoding a peptide or protein comprises or constitutes at least one amino acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the immunogenic fragments or variants of SEQ ID NO: 124 to SEQ ID NO: 254, SEQ ID NO: 546 to SEQ ID NO: 650, SEQ ID NO: 756 to 763, or any of these immunogenic fragments or variants.

[0614] 5. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-WDR72-2:4, said antigenic peptide comprising or constituting at least one amino acid sequence, said amino acid sequence being identical or having at least 80% identity with any one of SEQ ID NO: 124 to SEQ ID NO: 146, SEQ ID NO: 546 to SEQ ID NO: 571, SEQ ID NO: 756, SEQ ID NO: 757, or any immunogenic fragment or variant thereof.

[0615] 6. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-TRPC5-3:1, said antigenic peptide comprising or constituting at least one amino acid sequence, said amino acid sequence being identical or having at least 80% identity with any one of SEQ ID NO: 227 to SEQ ID NO: 237, SEQ ID NO: 630 to SEQ ID NO: 642, SEQ ID NO: 761, or any of these immunogenic fragments or variants.

[0616] 7. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-CLEC2D-9:1, said antigenic peptide comprising or constituting at least one amino acid sequence, said amino acid sequence being identical or having at least 80% identity with any one of SEQ ID NO: 238 to SEQ ID NO: 243, SEQ ID NO: 643 to SEQ ID NO: 650, SEQ ID NO: 762, or any immunogenic fragment or variant thereof.

[0617] 8. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by KCNMB2-AS1:4, said antigenic peptide comprising or constituting at least one amino acid sequence that is identical or has at least 80% identity with any one of SEQ ID NO: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, SEQ ID NO: 758, or any of these immunogenic fragments or variants.

[0618] 9. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-ZC3H8-6:1, said antigenic peptide comprising or constituting at least one amino acid sequence that is identical or has at least 80% identity with any one of SEQ ID NO: 162 to SEQ ID NO: 168, SEQ ID NO: 590 to SEQ ID NO: 603, SEQ ID NO: 759, or any immunogenic fragment or variant thereof.

[0619] 10. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-NTF3-5:5, said antigenic peptide comprising or constituting at least one amino acid sequence that is identical or has at least 80% identity with any one of SEQ ID NO: 169 to SEQ ID NO: 226, SEQ ID NO: 604 to SEQ ID NO: 629, SEQ ID NO: 760, or any immunogenic fragment or variant thereof.

[0620] 11. The artificial nucleic acid according to claims 1 to 4, comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by LINC00893:25, said antigenic peptide comprising or constituting at least one amino acid sequence that is identical to or has at least 80% identity with any one of SEQ ID NO: 244 to SEQ ID NO: 254, SEQ ID NO: 763, or any of these immunogenic fragments or variants.

[0621] 12. The artificial nucleic acid according to claims 1 to 11, wherein the at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from lncRNA-encoded peptides or proteins, wherein the nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 318 to SEQ ID NO: 448, SEQ ID NO: 651 to SEQ ID NO: 755, SEQ ID NO: 764 to SEQ ID NO: 771, or any fragments or variants thereof.

[0622] 13. The artificial nucleic acid according to Clause 12, wherein the at least one coding sequence comprises a GC-optimized nucleic acid sequence encoding at least one antigenic peptide selected from lncRNA-encoded peptides or proteins, wherein the nucleic acid sequence is identical or has at least 80% identity with any one of SEQ ID NO: 318 to SEQ ID NO: 448, SEQ ID NO: 651 to SEQ ID NO: 755, SEQ ID NO: 764 to SEQ ID NO: 771, or any fragments or variants thereof.

[0623] 14. The artificial nucleic acid according to Clause 1, wherein the tumor neoantigen is selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, PIK3CA neoantigen, NFE2L2 neoantigen, or CNOT9 neoantigen, or immunogenic fragments or variants thereof.

[0624] 15. The artificial nucleic acid according to Clause 14, wherein at least one antigenic peptide selected from a peptide or protein of a tumor neoantigen contains at least one T-cell epitope, preferably at least one CD8+ T-cell epitope.

[0625] 16. The artificial nucleic acid according to Clause 14 or 15, wherein at least one antigenic peptide selected from a tumor neoantigen peptide or protein comprises or constitutes at least one amino acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 255 to SEQ ID NO: 317 or any of these immunogenic fragments or immunogenic variants.

[0626] 17. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of NRAS tumor neoantigens, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:267 to SEQ ID NO:272.

[0627] 18. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of the TYW1B tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO: 273, SEQ ID NO: 274.

[0628] 19. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of ECPAS tumor neoantigens, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with SEQ ID NO: 275.

[0629] 20. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of the MAP2K1 tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:276 or SEQ ID NO:277.

[0630] 21. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of the TOMM22 tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:278 or SEQ ID NO:279.

[0631] 22. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of the GLB1 tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO: 280, SEQ ID NO: 281.

[0632] 23. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of MAGE-A3 tumor neoantigens, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or an immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:282 or SEQ ID NO:283.

[0633] 24. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of the ATAD2 tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO: 284 to SEQ ID NO: 286.

[0634] 25. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of BRAF tumor neoantigens, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:262 to SEQ ID NO:266.

[0635] 26. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of EGFR tumor neoantigens, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:255 to SEQ ID NO:257.

[0636] 27. The artificial nucleic acid according to claims 14 to 16, comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of TP53 tumor neoantigen, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or variant thereof, the amino acid sequence being identical to or having at least 80% identity with any one of SEQ ID NO:258 to SEQ ID NO:261.

[0637] 28. The artificial nucleic acid according to claims 14 to 27, wherein the at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins of tumor neoantigens, wherein the nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants.

[0638] 29. The artificial nucleic acid according to Clause 28, wherein the at least one coding sequence comprises a GC-optimized nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins of tumor neoantigens, wherein the nucleic acid sequence is identical to or has at least 80% identity with any one of SEQ ID NO: 449 to SEQ ID NO: 511, or any fragment thereof.

[0639] 30. The artificial nucleic acid according to any one of the preceding clauses, wherein the at least one coding sequence encodes 2 to 10 different tumor antigens, preferably 2 to 5 different tumor antigens.

[0640] 31. The artificial nucleic acid according to Clause 30, wherein the different tumor antigens are separated by a connector element.

[0641] 32. The artificial nucleic acid according to any one of the preceding clauses, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one immune response activation signal transduction protein.

[0642] 33. The artificial nucleic acid according to any one of the preceding clauses, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one T helper cell epitope.

[0643] 34. The artificial nucleic acid according to any one of the preceding clauses, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one signal peptide.

[0644] 35. The artificial nucleic acid according to any one of the preceding clauses, wherein the at least one coding sequence is a codon-modified coding sequence, preferably wherein the codon-modified coding sequence is selected from C-maximized coding sequences, CAI-maximized coding sequences, human codon selection-adapted coding sequences, G / C content-modified coding sequences, and G / C optimized coding sequences, or any combination thereof.

[0645] 36. The artificial nucleic acid according to Clause 35, wherein the at least one codon-modified coding sequence is a G / C-optimized coding sequence.

[0646] 37. The artificial nucleic acid according to any one of the preceding clauses, wherein the nucleic acid comprises at least one untranslated region (UTR), preferably selected from at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR.

[0647] 38. The artificial nucleic acid according to Clause 37, wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence selected from or derived from the 3'-UTR of a gene selected from: PSMB3, ALB7, α-globin, β-globin, ANXA4, CASP1, COX6B1, FIG4, GNAS, NDUFA1, RPS9, SLC7A3 or TUBB4B, or a nucleic acid sequence of a homology, fragment or variant of any of these genes, preferably wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with any of these fragments or variants.

[0648] 39. The artificial nucleic acid according to clause 37 or 38, wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence selected from or derived from the 3'-UTR of the PSMB3 gene, wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 112 to SEQ ID NO: 123, preferably SEQ ID NO: 67, or any of these fragments or variants.

[0649] 40. The artificial nucleic acid according to Clause 37, wherein the at least one heterologous 5'-UTR comprises or constitutes a nucleic acid sequence selected from or derived from the 5'-UTR of a gene selected from: HSD17B4, RPL32, AIG1, α-globin, ASAH1, ATP5A1, COX6C, DPYSL2, MDR, MP68, NDUFA4, NOSIP, RPL31, RPL35A, SLC7A3, TUBB4B, or UBQLN2, or a nucleic acid sequence of a homolog, fragment, or variant of any of these genes, preferably wherein the at least one heterologous 5'-UTR comprises or constitutes a nucleic acid sequence with SEQ ID NO: 12 to SEQ ID NO: 45, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67 ... NO:65, or any of these fragments or variants, are identical or have at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity nucleic acid sequences.

[0650] 41. The artificial nucleic acid according to clause 37 or 40, wherein the at least one heterologous 5'-UTR comprises or constitutes a nucleic acid sequence selected from or derived from the 5'-UTR of the HSD17B4 gene, wherein the at least one heterologous 5'-UTR comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a fragment or variant of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 64, SEQ ID NO: 65, preferably SEQ ID NO: 13, or any of these.

[0651] 42. The artificial nucleic acid according to clauses 37 to 41, wherein the at least one heterologous 5'-UTR is selected from HSD17B4 and the at least one heterologous 3' UTR is selected from PSMB3.

[0652] 43. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is selected from DNA or RNA, preferably RNA.

[0653] 44. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is an RNA selected from mRNA, circular RNA, replicon RNA or viral RNA.

[0654] 45. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is mRNA.

[0655] 46. ​​The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid, preferably the RNA, comprises at least one poly(A) sequence, preferably wherein the at least one poly(A) sequence comprises about 40 to about 500 adenosine nucleotides.

[0656] 47. The artificial nucleic acid according to Clause 46, wherein the at least one poly(A) sequence comprises about 60 to about 150 adenosine nucleotides, preferably about 100 adenosine nucleotides.

[0657] 48. The artificial nucleic acid according to clause 46 or 47, wherein the at least one poly(A) sequence is located at the 3' end, optionally wherein the 3' terminal nucleotide is adenosine.

[0658] 49. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid, preferably the RNA, comprises at least one poly(C) sequence and / or at least one miRNA binding site and / or at least one histone stem-loop sequence.

[0659] 50. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid, preferably the RNA, comprises at least one histone stem-loop sequence, wherein the histone stem-loop sequence comprises or constitutes a nucleic acid sequence, or a fragment or variant thereof, that is identical to or has at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 3 or SEQ ID NO: 4.

[0660] 51. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is RNA, the RNA comprising at least one modified nucleotide, preferably a modified nucleotide selected from pseudouridine (ψ) or N1-methylpseudouridine (m1ψ).

[0661] 52. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is a modified RNA, wherein each uracil is replaced by a modified nucleotide.

[0662] 53. The artificial nucleic acid according to clause 51 or 52, wherein the modified nucleotide is N1-methylpseuuridine (m1ψ).

[0663] 54. The artificial nucleic acid as described in clauses 1 to 50, wherein the nucleic acid is RNA that does not contain modified nucleotides.

[0664] 55. The artificial nucleic acid according to any of the preceding claims, wherein the artificial nucleic acid is RNA containing a 5'-cap structure.

[0665] 56. The artificial nucleic acid according to Clause 55, wherein the 5'-cap structure is selected from the cap1 structure or a modified cap1 structure.

[0666] 57. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is RNA transcribed in vitro.

[0667] 58. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid is purified RNA, preferably wherein the RNA has been purified by at least one step of RP-HPLC, AEX, SEC, hydroxyapatite chromatography, TFF, filtration, precipitation, microsphere flow-through chromatography, oligo(dT) purification, cellulose-based purification, or any combination thereof.

[0668] 59. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid, preferably the RNA, has at least 50%, preferably at least 60%, more preferably at least 70%, and most preferably at least 80% integrity.

[0669] 60. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid, preferably the RNA, is suitable for treating or preventing diseases, disorders or conditions, preferably tumor diseases, disorders or conditions.

[0670] 61. The artificial nucleic acid according to any one of the preceding clauses, wherein the artificial nucleic acid, preferably the RNA, comprises the following sequence elements, preferably in the 5' to 3' orientation:

[0671] A) 5'-cap structure;

[0672] B) 5'-UTR, preferably selected from the 5'-UTR of the HSD17B4 gene;

[0673] C) A coding sequence that encodes at least one tumor antigen;

[0674] D) 3'-UTR, preferably selected from the 3'-UTR of the PSMB3 gene;

[0675] E) Optionally, histone stem-loop; and

[0676] F) poly(A) sequence, which preferably contains about 100 A nucleotides.

[0677] 62. The artificial nucleic acid according to claims 1 to 61, wherein said artificial nucleic acid is mRNA and comprises the following sequence elements in the 5' to 3' orientation:

[0678] A) 5'-cap structure;

[0679] B) 5'-UTR, which contains or constitutes a nucleic acid sequence, or a fragment or variant thereof, that is identical to or has at least 80% identity with SEQ ID NO: 13;

[0680] C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from long non-coding RNA (lncRNA) encoded by at least one antigenic peptide or protein, wherein the nucleic acid sequence is identical or has at least 80% identity with any one of SEQ ID NO: 318 to SEQ ID NO: 448, SEQ ID NO: 651 to SEQ ID NO: 755, SEQ ID NO: 764 to SEQ ID NO: 771, or any fragment or variant thereof;

[0681] D) 3'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 80% identity with SEQ ID NO: 67, or a fragment or variant thereof;

[0682] E) Optionally, histone stem-loop; and

[0683] F) poly(A) sequence, which preferably contains about 100 A nucleotides.

[0684] 63. The artificial nucleic acid according to claims 1 to 61, wherein said artificial nucleic acid is mRNA and contains the following sequence elements in the 5' to 3' orientation:

[0685] A) 5'-cap structure;

[0686] B) 5'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 80% identity with SEQ ID NO: 13, or a fragment or variant thereof;

[0687] C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from a tumor neoantigen, wherein the nucleic acid sequence is identical or has at least 80% identity with any one of SEQ ID NO: 449 to SEQ ID NO: 511, or any fragment or variant thereof.

[0688] D) 3'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 80% identity with SEQ ID NO: 67, or a fragment or variant thereof;

[0689] E) Optionally, histone stem-loop; and

[0690] F) poly(A) sequence, which preferably contains about 100 A nucleotides.

[0691] 64. A pharmaceutical composition comprising at least one artificial nucleic acid, said artificial nucleic acid comprising at least one coding sequence encoding at least one tumor antigen as defined in any one of clauses 1 to 63.

[0692] 65. The pharmaceutical composition according to Clause 64, comprising a plurality of artificial nucleic acids, each of which encodes at least one different tumor antigen.

[0693] 66. The pharmaceutical composition according to clause 64 or 65, wherein the at least one artificial nucleic acid, preferably the RNA, is formulated in at least one cationic compound or polycationic compound.

[0694] 67. The pharmaceutical composition according to Clause 66, wherein the at least one cationic compound or polycationic compound is selected from cationic polymers or polycationic polymers, cationic polysaccharides or polycationic polysaccharides, cationic lipids or polycationic lipids, cationic proteins or polycationic proteins, cationic peptides or polycationic peptides, or any combination thereof.

[0695] 68. The pharmaceutical composition according to clauses 64 to 67, wherein the at least one artificial nucleic acid, preferably the RNA, is formulated in a lipid-based carrier.

[0696] 69. The pharmaceutical composition according to Clause 68, wherein the lipid-based carrier is selected from liposomes, lipid nanoparticles, lipid complexes, solid lipid nanoparticles, lipid-polymer complexes, and / or nanoliposomes.

[0697] 70. The pharmaceutical composition according to clause 68 or 69, wherein the lipid-based carrier is a lipid nanoparticle.

[0698] 71. The pharmaceutical composition according to clauses 68 to 70, wherein the lipid-based carrier comprises at least one lipid that reduces aggregation, at least one cationic lipid or ionizable lipid, at least one neutral lipid or phospholipid, and at least one steroid or steroid analogue.

[0699] 72. The pharmaceutical composition according to Clause 71, wherein the lipid that reduces aggregation is a polymer-conjugated lipid selected from PEG-conjugated lipids or lipids without PEG.

[0700] 73. The pharmaceutical composition according to Clause 72, wherein the polymer-conjugated lipid is selected from DMG-PEG2000, C10-PEG2K, Cer8-PEG2K or POZ lipids.

[0701] 74A. The pharmaceutical composition according to clauses 71 to 73, wherein the cationic lipid or ionizable lipid is selected from amino lipids, preferably wherein the amino lipid contains a tertiary amine group.

[0702] 74B. The pharmaceutical composition according to clauses 71 to 74A, wherein the cationic lipid or ionizable lipid is selected from or derived from formula (III-1, as defined herein), preferably wherein one of L1 or L2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -SS-, -C(=O)S-, -SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, -NRaC(=O)NRa-, -OC(=O)NRa- Or -NRaC(=O)O-, and the other of L1 or L2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -SS-, -C(=O)S-, -SC(=O-, -NRaC(=O)-, -C(=O)NRa-, -NRaC(=O)NRa-, -OC(=O)NRa- or -NRaC(=O)O- or directly bonded; G1 and G2 are each independently unsubstituted C1 to C12 alkylene oxides. R1 is a C1 to C12 alkenyl group; R2 is a C1 to C24 alkenyl group, C1 to C24 alkenyl group, C3 to C8 cycloalkyl group, C3 to C8 cycloalkyl group; R3 is H or C1 to C12 alkenyl group; R4 is a C1 to C12 alkenyl group; R5 is H or C1 to C24 alkenyl group; and x is 0, 1 or 2.

[0703] 74C. The pharmaceutical composition according to clauses 71 to 74A or 74B, wherein the cationic lipid is selected from or derived from formula III-3 of WO2018078053, preferably wherein the cationic lipid has the chemical name ((4-hydroxybutyl)azadiyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate), also known as ALC-0315.

[0704] 75. The pharmaceutical composition according to clauses 71 to 74A or 74B or 74C, wherein the cationic lipid or ionizable lipid is selected from ALC-0315, SM-102, SS-33 / 4PE-15, HEXA-C5DE-PipSS or compound C26.

[0705] 76. The pharmaceutical composition according to clauses 71 to 75, wherein the neutral lipid or phospholipid is selected from DSPC, DHPC, or DPhyPE.

[0706] 77. The pharmaceutical composition according to clauses 71 to 76, wherein the steroid or steroid analogue is selected from cholesterol, cholesterol hemisuccinate (CHEMS), preferably cholesterol.

[0707] 78. The pharmaceutical composition according to clauses 68 to 77, wherein the lipid-based carrier, preferably the LNP, comprises

[0708] (i) at least one cationic lipid, preferably as defined in clauses 74 or 75;

[0709] (ii) At least one neutral lipid, preferably as defined in Clause 76;

[0710] (iii) at least one steroid or steroid analogue, preferably as defined in clause 77; and

[0711] (iv) At least one lipid that reduces aggregation, preferably as defined in clauses 72 to 73.

[0712] 79. The pharmaceutical composition according to clauses 68 to 78, wherein the lipid-based carrier comprises about 20% to 60% cationic lipids, about 5% to 25% neutral lipids, about 25% to 55% steroids or steroid analogs, and about 0.5% to 15% lipids that reduce aggregation.

[0713] 80. The pharmaceutical composition according to claims 68 to 79, wherein in the lipid-based carrier, the weight / weight ratio of lipid to nucleic acid (e.g., RNA) is from about 10:1 to about 60:1.

[0714] 81. The pharmaceutical composition according to clauses 68 to 80, wherein the N / P ratio of the lipid-based carrier encapsulating the nucleic acid, preferably the RNA, is from about 1 to about 20.

[0715] 82. The pharmaceutical composition according to clauses 68 to 81, wherein the lipid-based carrier has a Z-average particle size of about 50 nm to about 200 nm.

[0716] 83. The pharmaceutical composition according to claims 64 to 82, further comprising at least one antagonist of at least one RNA-sensing pattern recognition receptor selected from Toll-like receptor antagonists, preferably a TLR7 antagonist and / or a TLR8 antagonist, most preferably RNA according to SEQ ID NO: 85 of WO2021028439.

[0717] 84. The pharmaceutical composition or artificial nucleic acid according to any one of the preceding clauses, wherein the composition or nucleic acid, after being administered intramuscularly, intratumorally or intravenously to a cell, tissue or object, generates an encoded tumor antigen.

[0718] 85. A pharmaceutical composition or artificial nucleic acid according to any one of the preceding clauses, wherein, after administration of the composition or nucleic acid intramuscularly, intratumorally, or intravenously to a cell, tissue, or object, it generates an encoded tumor antigen and induces epitope-specific CD8+ T cells in the object.

[0719] 86. A tumor antigen or a composition of at least one tumor antigen, wherein the tumor antigen is characterized by any of the preceding clauses, preferably by any of clauses 2 to 4 or clauses 14 to 16.

[0720] 87. A tumor antigen or composition according to Clause 86, wherein the tumor antigen comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 124 to SEQ ID NO: 254, SEQ ID NO: 546 to SEQ ID NO: 650, SEQ ID NO: 756 to SEQ ID NO: 763.

[0721] 88. A tumor antigen or composition according to Clause 86, wherein the tumor antigen comprises or constitutes at least one amino acid sequence, or any immunogenic fragment or immunogenic variant thereof, said amino acid sequence being identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 255 to SEQ ID NO: 317.

[0722] 89. A tumor antigen or composition according to clauses 86 to 88, wherein the composition comprises a plurality of tumor antigens, and optionally, wherein the antigen is formulated in a pharmaceutically acceptable carrier.

[0723] 90. An antibody, T cell, or TCR that has been generated or stimulated with any of the tumor antigens or nucleic acids as defined in any of the preceding clauses.

[0724] 91. A combination comprising at least two or more treatment modalities, said treatment modalities being selected from...

[0725] a) at least one artificial nucleic acid according to clauses 1 to 62; and / or

[0726] b) At least one pharmaceutical composition according to clauses 63 to 85; and / or

[0727] c) At least one tumor antigen or composition according to clauses 86 to 89; and / or

[0728] d) At least one antibody, T cell or TCR according to clause 90.

[0729] 92. A kit or kit comprising at least one artificial nucleic acid according to any one of Clauses 1 to 62, and / or at least one pharmaceutical composition according to any one of Clauses 63 to 85, and / or at least one tumor antigen or composition of tumor antigens according to Clauses 86 to 89, and / or at least one antibody, T cell or TCR according to Clause 90, optionally comprising a liquid carrier for dissolution, and optionally comprising technical instructions providing information on the administration and dosage of the components.

[0730] 93. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a pharmaceutical agent.

[0731] 94. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine for treating or preventing cancer or any cancer-related disease, disorder or condition in a subject.

[0732] 95. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine for treating or preventing squamous cell carcinoma or any disease, disorder or condition associated with squamous cell carcinoma in a subject, preferably wherein the squamous cell carcinoma is selected from LUSC / squamous non-small cell lung cancer (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC).

[0733] 96. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine for treating or preventing NSCLC or any disease, disorder or condition associated with NSCLC in a subject.

[0734] 97. A medicament for the treatment or prevention of NSCLC as described in Clause 96, wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises (a) at least one antigenic peptide of a peptide or protein encoded by lnc-WDR72-2:4, lnc-TRPC5-3:1, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, or an immunogenic fragment or variant of any of these; and / or (b) at least one antigenic peptide of a peptide or protein selected from BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, PIK3CA neoantigen, NFE2L2 neoantigen, preferably selected from EGFR neoantigen or TP53 neoantigen, or an immunogenic fragment or variant of any of these.

[0735] 96B. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or tumor antigen composition according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine for the treatment or prevention of HNSCC or any disease, disorder or condition associated with HNSCC in a subject.

[0736] 97B. Use as a medicament for the treatment or prevention of HNSCC as described in Clause 96B, wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises at least one antigenic peptide selected from lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5 or lnc-ZC3H8-6:1, or any immunogenic fragment or variant of these.

[0737] 98. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine for treating or preventing melanoma or any disease, disorder or condition associated with melanoma in a subject.

[0738] 99. Use as a medicament for the treatment or prevention of melanoma as described in Clause 98, wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises (a) at least one antigenic peptide selected from lnc-CLEC2D-9:1, lnc-TRPC5-3:1, or LINC00893:25 encoding a peptide or protein, or any immunogenic fragment or variant thereof; and / or (b) selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A 3. At least one antigenic peptide of a peptide or protein of a neoantigen, ATAD2 neoantigen, BRAF neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, or CNOT9 neoantigen, preferably at least one antigenic peptide selected from at least one antigenic peptide of an immunogenic fragment or variant of NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, or BRAF neoantigen, or any of these.

[0739] 99B. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, for use in the treatment or prevention of adrenocortical carcinoma, bladder urothelial carcinoma, invasive breast cancer, cervical cancer (cervical squamous cell carcinoma and adenocarcinoma), colonic adenocarcinoma, renal papillary cell carcinoma, or lung adenocarcinoma. Drugs for lymphoid tumors, diffuse large B-cell lymphoma, rectal adenocarcinoma, sarcoma, cutaneous melanoma, thymoma, thyroid cancer, hepatocellular carcinoma, uterine carcinosarcoma, or endometrial cancer, or any related diseases, disorders, or conditions, wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-ZC3H8-6:1, or an immunogenic fragment or variant thereof.

[0740] 99C. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicament for the treatment or prevention in a subject of bladder urothelial carcinoma, cervical cancer (cervical squamous cell carcinoma and cervical adenocarcinoma), thymoma, or endometrial cancer or any related disease, wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit comprises at least one antigenic peptide selected from a peptide or protein encoded by KCNMB2-AS1:4, or an immunogenic fragment or variant thereof.

[0741] 99D. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine in a subject to treat or prevent ovarian serous cystadenocarcinoma, testicular germ cell tumor, thymoma or uterine carcinosarcoma, or any related disease, wherein the at least one tumor antigen provided by said artificial nucleic acid, said pharmaceutical composition, said tumor antigen or a composition of tumor antigens, said combination, or said kit or kit comprises at least one antigenic peptide selected from lnc-NTF3-5:5 encoded by lnc-NTF3-5:5, or an immunogenic fragment or variant thereof.

[0742] 99E. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicament for the treatment or prevention in a subject of bladder urothelial carcinoma, colon adenocarcinoma, esophageal cancer or gastric adenocarcinoma or any related disease, wherein the at least one tumor antigen provided by said artificial nucleic acid, said pharmaceutical composition, said tumor antigen or a composition of tumor antigens, said combination, or said kit or kit comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-WDR72-2:4, or an immunogenic fragment or variant thereof.

[0743] 99F. An artificial nucleic acid according to any one of Clauses 1 to 62, a pharmaceutical composition according to any one of Clauses 63 to 85, a tumor antigen or a composition of tumor antigens according to Clauses 86 to 89, an antibody, T cell or TCR according to Clause 90, a combination according to Clause 91, or a kit or kit according to Clause 92, used as a medicine for treating or preventing cutaneous melanoma, testicular germ cell tumor or uveal melanoma, or any related disease, disorder or condition in a subject, wherein the at least one tumor antigen provided by said artificial nucleic acid, said pharmaceutical composition, said tumor antigen or a composition of tumor antigens, said combination, or said kit or kit comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-CLEC2D-9:1, or an immunogenic fragment thereof.

[0744] 100. Use as a medicine as described in clauses 93 to 99F, wherein the medicine is administered to the subject via intramuscular, intratumoral, or intravenous administration.

[0745] 101. A method for treating or preventing a disease, disorder, or symptom, the method comprising applying or administering to a subject in need an effective amount of an artificial nucleic acid as described in Clauses 1 to 62, a pharmaceutical composition as described in Clauses 63 to 85, a tumor antigen or a composition of tumor antigens as described in Clauses 86 to 89, an antibody, T cell, or TCR as described in Clause 90, a combination as described in Clause 91, or a kit or kit as described in Clause 92.

[0746] 102. The method described in Clause 101, wherein the disease, disorder or ailment is cancer.

[0747] 103. The method according to Clause 102, wherein the cancer is squamous cell carcinoma, preferably selected from squamous non-small cell lung cancer (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC).

[0748] 104. The method according to Clause 102 or 103, wherein the cancer is NSCLC, and wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, or the kit or kit kit comprises (a) at least one antigenic peptide encoded by a peptide or protein, or an immunogenic fragment or variant of any of lnc-WDR72-2:4, lnc-TRPC5-3:1, KCNMB2-AS1:4, lnc-NTF3-5:5 or lnc-ZC3H8-6:1; and / or (b) at least one antigenic peptide selected from peptides or proteins of BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, PIK3CA neoantigen or NFE2L2 neoantigen, preferably selected from at least one antigenic peptide of EGFR neoantigen or TP53 neoantigen, or an immunogenic fragment or variant of any of these.

[0749] 105. The method according to Clause 102, wherein the cancer is melanoma, and wherein the at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, or the kit or kit kit comprises (a) at least one antigenic peptide selected from lnc-CLEC2D-9:1 or lnc-TRPC5-3:1 encoding a peptide or protein, or any of these immunogenic fragments or variants; and / or (b) selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD 2. At least one antigenic peptide of a peptide or protein of a neoantigen, BRAF neoantigen, DIP2B neoantigen, TRRAP neoantigen, RAC1 neoantigen, AP4B1 neoantigen, VWA5A neoantigen, CLTC neoantigen, RPL9 neoantigen, RPAP1 neoantigen, RPE65 neoantigen, PATZ1 neoantigen, or CNOT9 neoantigen, preferably selected from at least one antigenic peptide of an immunogenic fragment or variant of NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, or BRAF neoantigen, or any of these.

[0750] 106. The method described in accordance with Clause 101 is characterized by any of the medical use features as defined in Clauses 93 to 100. Attached Figure Description

[0751] Figure 1An exemplary neoantigen identification workflow is illustrated. *Immunogenicity prediction: neoIM; MHC presentation prediction: MHCnuggets & neoMS. **Considerations include host gene expression, sample coverage, HLA coverage / distribution, MHC presentation capacity, and immunogenicity.

[0752] Figure 2 The study demonstrated the high tumor specificity of the lncRNA transcript lnc-CLEC2D-9:1 (corresponding to smORF: lncCLEC2D91412616). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in melanoma, while it was lowly expressed to absent in healthy tissues (except for the immune-exempt tissue testis).

[0753] Figure 3 The study demonstrated the high tumor specificity of the lncRNA transcript lnc-TRPC5-3:1 (corresponding to smORF: lncTRPC53116031801). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in melanoma, while it was lowly expressed to absent in healthy tissues.

[0754] Figure 4 The study demonstrated the high tumor specificity of the lncRNA transcript lnc-TRPC5-3:1 (corresponding to smORF: lncTRPC53116031801). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in NSCLC (lung adenocarcinoma, LUAD, and lung squamous cell carcinoma, LUSC), while it was lowly expressed to absent in healthy tissues.

[0755] Figure 5 The study demonstrated the high tumor specificity of the lncRNA transcript lnc-WDR72-2:4 (corresponding to smORFs: lncWDR722415641696 and lncWDR722416421696). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in NSCLC (lung squamous cell carcinoma, LUSC), while it was lowly expressed to absent in healthy tissues.

[0756] Figure 6The study demonstrated high tumor specificity for the lncRNA transcript KCNMB2-AS1:4 (corresponding to smORF: KCNMB2AS14633753). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in NSCLC (lung squamous cell carcinoma, LUSC), while it was expressed at low levels to absent expression in healthy tissues.

[0757] Figure 7 The study demonstrated high tumor specificity for the lncRNA transcript lnc-NTF3-5:5 (corresponding to smORF: lncNTF35511951417). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in NSCLC (lung squamous cell carcinoma, LUSC), while it was lowly expressed to absent in healthy tissues.

[0758] Figure 8 The study demonstrated the high tumor specificity of the lncRNA transcript lnc-ZC3H8-6:1 (corresponding to smORF: lncZC3H86163144). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases showed that the target was highly expressed in NSCLC (lung adenocarcinoma, LUAD, and lung squamous cell carcinoma, LUSC), while it was lowly expressed to absent in healthy tissues.

[0759] Figure 9 The translation of the peptide of smORF lncTRPC53116031801 is shown to be nearly absent in healthy tissues. Mass spectrometry data show that the level is very low or even absent (asterisk) in various healthy tissues compared to different high-abundance controls (e.g., housekeeping gene; dot).

[0760] Figure 10 In vitro antigen validation of the melanoma neoantigen (peptide pool 1) in a DC-T cell assay is illustrated (see Example 6.1, Tables E3-E5 for further details). Following two rounds of stimulation (R1 and R2), pan-T cell IFN-γ readings were performed on 10 PBMC donors (D1-D10). Seven donors showed antigen-specific T cell responses to at least one peptide from peptide pool 1. Recall and memory responses were observed in one donor and one donor, respectively.

[0761] Figure 11In vitro antigen validation of the melanoma neoantigen (peptide pool 2) in a DC-T cell assay is shown (see Example 6.1, Tables E3-E5 for further details). Following two rounds of stimulation (R1 and R2), pan-T cell IFN-γ readings were performed on 10 PBMC donors (D1-D10). Seven donors showed antigen-specific T cell responses to at least one peptide from peptide pool 2. Memory responses were observed in two donors.

[0762] Figure 12 In vitro antigen validation of the melanoma cancer antigen (peptide pool 3) in a DC-T cell assay is shown (see Example 6.1, Tables E3-E5 for further details). Following two rounds of stimulation (R1 and R2), pan-T cell IFN-γ readings were performed on 10 PBMC donors (D1-D10). All donors showed antigen-specific T cell responses to at least one peptide from peptide pool 3. Recall and memory responses were observed in 4 and 4 donors, respectively.

[0763] Figure 13 In vitro antigen validation of the melanoma neoantigen (peptide pool 4) in a DC-T cell assay is shown (see Example 6.1, Tables E3-E5 for further details). Following two rounds of stimulation (R1 and R2), pan-T cell IFN-γ readings were performed on 10 PBMC donors (D1-D10). Eight donors showed antigen-specific T cell responses to at least one peptide from peptide pool 4. Recall and memory responses were observed in two and one donor, respectively.

[0764] Figure 14 In vitro antigen validation of the melanoma neoantigen (peptide pool 5) in a DC-T cell assay is shown (see Example 6.1, Tables E3-E5 for further details). Following two rounds of stimulation (R1 and R2), pan-T cell IFN-γ readings were performed on 10 PBMC donors (D1-D10). Four donors showed antigen-specific T cell responses to peptide pool 5. Recall and memory responses were observed in one and two donors, respectively.

[0765] Figure 15 Example 6.1 / is shown. Figures 10 to 14 An overview of all recall and memory responses observed.

[0766] Figure 16 The in vitro antigen validation of the specified smORF in the DC-T cell assay is shown (see Example 6.2, Tables E6 through E8 for further details). IFN-γ readings of pan-T cells were performed on each PBMC donor after two rounds of stimulation (R1 and R2).

[0767] Figure 17In vitro antigen validation of the specified melanoma neoantigen in a DC-T cell assay is shown (see Example 6.3, Tables E9 to E10A for further details). IFN-γ readings of pan-T cells were performed on each PBMC donor after two rounds of stimulation (R1 and R2).

[0768] Figure 18 In vivo immunogenicity of the LNP-formulated mRNA vaccine R12300, encoding two smORFs (one of which is expressed in melanoma, etc.), in CB6F1 hybrid mice. Mice were intramuscularly immunized with 5 µg of LNP-formulated R12300 on days 0, 7, and 14. Immunization with an unrelated mRNA (PpLuc; R8730) served as a control. On day 21, mice were sacrificed and spleen cells were isolated. Spleen cells were restimulated with a 15-mer peptide library targeting lncCLEC2D91412616 or DMSO as controls, and analyzed by flow cytometry. The intensity of CD8+ T cell responses (top) and CD4+ T cell responses (bottom) against the encoded smORF peptide lncCLEC2D91412616 is shown as the percentage of IFN-γ+TNF+ cells in either CD8+ T cells or CD4+ T cells. Median values ​​were plotted.

[0769] Figure 19 The in vivo immunogenicity of the LNP-formulated mRNA vaccine R12301, encoding multiple smORFs (including those expressed in NSLSC, etc.), in CB6F1 hybrid mice is shown. Mice were intramuscularly immunized with 5 µg of LNP-formulated R12301 on days 0, 7, and 14. Immunization with unrelated mRNA (PpLuc; R8730) served as a control. On day 21, mice were sacrificed and spleen cells were isolated. Spleen cells were individually restimulated with a peptide library targeting each smORF (a 15-mer library covering the complete antigen) or DMSO as controls, and analyzed by flow cytometry. The intensity of CD8+ T cell responses (top) and CD4+ T cell responses (bottom) to four encoded smORF peptides, KCNMB2AS14633753, lncNTF35511951417, lncWDR722415641696, and lncZC3H86163144 (NKZW-smORF), is shown as the percentage of IFN-γ+TNF+ cells in either CD8+ T cells or CD4+ T cells. Median values ​​are plotted.

[0770] Example:

[0771] The following are examples illustrating various embodiments of the invention. However, the scope of the invention should not be limited to the specific embodiments presented herein, but should be understood to be applicable to other compositions or uses, such as those defined in this specification. Therefore, the following preparations and examples are given to enable those skilled in the art to more clearly understand and practice the invention. In fact, various modifications to the invention, in addition to those described herein, will become readily apparent to those skilled in the art from the foregoing description, the drawings, and the following examples.

[0772] Example 1: Antigen Discovery and Screening Methods

[0773] The aim of this embodiment was to identify cancer antigens that generate epitopes in patients with NSCLC or melanoma. The analysis focused on two classes of targets: tumor-specific antigens (novel epitopes) derived from somatic mutations and tumor-associated antigens encoded by small open reading frames (smORFs) identified in long non-coding RNAs (lncRNAs). Using transcriptomics and proteomics, the identified smORFs / lncRNAs showed clear tumor-associated expression and were detected at low levels only in normal tissues. For both classes of antigens, candidates were selected using major histocompatibility complex (MHC) binding and immunogenicity prediction.

[0774] Discovery of new epitopes:

[0775] Tumor-specific variants were detected using sequence alignment map (BAM) files from the Cancer Genome Atlas (TCGA) database. A combination of parameters was used to select the most promising candidates. An immunogenicity score (neoim_score, with larger values ​​reflecting a higher probability of immunogenicity) was used with a threshold of 0.55. Furthermore, only genes showing an expression level of at least 20 transcripts per million (TPM) were considered. Finally, novel epitopes were prioritized based on their expected population coverage (the proportion of patients with mutations and predicted MHC-I binding alleles). The neoantigen identification workflow is as follows: Figure 1 As shown.

[0776] Discovery of smORF in lncRNAs:

[0777] To identify smORFs within lncRNAs, coding potential within lncRNA sequences was first predicted. Then, differential transcriptomic expression analysis was performed, comparing tumor and normal expression; only transcripts with a log2 (fold change) greater than 2 and an adjusted p-value less than 1e-4 were considered for further analysis. Furthermore, MHC-I binding and immunogenicity of smORFs 8 to 11 nmers were predicted, and only those nmers predicted to bind to MHC-I with a neoim_score greater than 0.5 were retained. Further, only lncRNAs with translational evidence identified by mass spectrometry (PXD009630 - melanoma, and PXD002612 - lung cancer) were considered. As a final filtering step, lncRNAs / smORFs showing low expression in normal tissues were retained using transcriptomic analysis (GTEx V7) and mass spectrometry (PXD010154).

[0778] The immunogenicity score (neoim_score) is generated using a machine learning algorithm and reflects the probability that the epitope is immunogenic. RNA-seq expression is given as TPM; the TPM normalizes the RNA-seq sequence fragments by the length and sequencing depth of each gene and is typically used to compare expression levels across different samples. The expected coverage of the neoantigen is calculated by multiplying the population frequency of the mutation by the predicted frequency of the HLA alleles binding to that epitope. HLA allele prediction is performed using a machine learning algorithm.

[0779] Example 2: Antigen discovery and selection of neoantigens for melanoma and NSCLC

[0780] The aim of this experiment was to discover cancer antigens in melanoma and NSCLC derived from tumor-specific neoantigens. The high-ranking hits are summarized in Table 3.

[0781] Table 3 shows the identified neoantigens for melanoma and NSCLC. For the corresponding neoantigens in the table, the RNAseq expression level (mean expression level), predicted immunogenicity score (neoim_ score), coverage in the corresponding cancer type (mean expected coverage), expected HLA allele binding (HLA mupexi allele), and cancer indication and subgroup (M = melanoma; M(BRAF) = BRAF-mutated melanoma; M(NRAS) = NRAS-mutated melanoma; N = NSCLC; LUSC = squamous cell carcinoma of the lung (sqNSCLC); LUAD = adenocarcinoma of the lung). In this context, M / melanoma also specifically refers to cutaneous melanoma (SKCM).

[0782] Table 3: Identified suitable neoantigens for melanoma and NSCLC

[0783]

[0784]

[0785]

[0786] Example 3: Discovery and screening of melanoma antigens and NSCLC antigens encoded by lncRNAs

[0787] The aim of this experiment was to discover melanoma and NSCLC cancer antigens derived from lncRNA-smORF. The high-ranking hits are summarized in Table 4.

[0788] Table 4 shows the identified melanoma and NSCLC antigens encoded by lncRNA transcripts. For the corresponding smORF in the table, the RNA-seq expression level (mean expression), predicted immunogenicity score (neoim_score), and cancer indication and subgroup are indicated (M = melanoma; N = NSCLC; LUSC = squamous cell carcinoma of the lung (sqNSCLC); LUAD = adenocarcinoma of the lung). In this context, M / melanoma also specifically refers to cutaneous melanoma (SKCM).

[0789] Table 4: Identified suitable melanoma antigens and NSCLC antigens encoded by lncRNA transcripts

[0790]

[0791] Furthermore, data from a mass spectrometry study including 50 melanoma patients (PXD009630 - melanoma) and 57 lung cancer patients (PXD002612 - lung adenocarcinoma) confirmed that the preferred smORF was indeed translated. In this context, Table 4A presents the patient frequencies of peptides whose smORF translation could be detected by mass spectrometry in tumor samples.

[0792] Table 4A: Frequency of lncRNA-smORFs identified in melanoma and NSCLC

[0793]

[0794] *Tumor mass spectrometry sample frequency: based on 57 samples for NSCLC (N) and 50 samples for melanoma (M).

[0795] Example 4: Optimizing the high tumor specificity of lncRNAs

[0796] The aim of this experiment was to compare and analyze the expression of preferred lncRNA transcripts (Table E1) in tumor and healthy tissues. To this end, RNA-seq expression data from the TCGA (tumor tissue) and GTEX (healthy tissue) databases were analyzed. In addition, mass spectrometry data were also analyzed.

[0797] result

[0798] The lncRNA transcripts lnc-CLEC2D-9:1 and lnc-TRPC5-3:1 were highly expressed in melanoma, while lnc-TRPC5-3:1, lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5, and lnc-ZC3H8-6:1 were highly expressed in NSCLC. All lncRNA transcripts showed only low to absent expression in healthy tissues. One exception was the observation of lnc-CLEC2D-9:1 expression in the testes; however, since the testes are an immune-immune site, this expression was considered negligible. Results are as follows... Figures 2 to 8 As shown, and summarized in Table E1.

[0799] Table E1: Expression of preferred lncRNA transcripts in melanoma / NSCLC and healthy tissues

[0800]

[0801] na, unavailable; LUSC, squamous cell carcinoma of the lung; LUAD, adenocarcinoma of the lung.

[0802] Furthermore, mass spectrometry analysis confirmed that the peptides translated from the preferred smORF were expressed at low to absent levels in healthy tissues. Figure 9 Exemplary data for lncTRPC53116031801 is shown.

[0803] Overall, the data indicate that preferred lncRNAs and their derived smORFs are highly and frequently expressed in tumors, while their expression is low or absent in healthy tissues.

[0804] Example 5: Expression of lncRNA transcripts in the thymus

[0805] The aim of this experiment was to analyze the expression of preferred lncRNA transcripts (Table E1) in the thymus. Typically, antigen expression in the thymus is associated with central immune tolerance; therefore, ideally, the expression of target antigens used in vaccines should be low to absent in the thymus. For this purpose, transcriptomic analysis was performed on publicly available datasets.

[0806] result

[0807] Exemplary data from the three datasets depicted in Table E2 show that lncRNA transcripts are expressed only at low levels to absently in the thymus. This suggests that smORFs derived from these lncRNA transcripts, such as the preferred smORFs described in Table E1, are less likely to develop central immune tolerance.

[0808] Table E2: Expression of preferred lncRNA transcripts in the thymus

[0809]

[0810] Example 6: Antigen Validation - In Vitro Immunogenicity

[0811] The purpose of this experiment was to analyze the in vitro immunogenicity of different melanoma and NSCLC cancer antigens derived from tumor-specific neoantigens and lncRNA-derived smORFs as described in Examples 1 to 5.

[0812] Peptides containing the target epitopes were synthesized according to standard procedures. The amino acid sequences of the peptides are given in the respective examples. Peptide pools (Examples 6.1 and 6.2) or single peptides (Examples 6.3 and 6.4) were used.

[0813] The immunogenicity of the test peptide was assessed using an in vitro DC-T cell assay. In this assay, cryopreserved PBMCs from multiple (healthy) donors of various HLA types were retrieved from cryopreservation, and monocytes were isolated. The cultured monocytes were differentiated into immature dendritic cells (iDCs) using a cytokine mixture (GM-CSF and IL-4). The iDCs were loaded with the test peptide and further matured into mature dendritic cells (mDCs) using CD40L. Subsequently, the mDCs were co-cultured with previously isolated autologous pan-T cells for 8 days in the presence of the cytokine mixture. During co-culture, the medium and cytokines were changed periodically. After 8 days, pan-T cells were harvested (round 1 = R1) and restimulated with peptide-loaded monocytes, and co-cultured again for 8 days in the presence of the cytokine mixture (round 2 = R2). After rounds 1 and 2, pan-T cells were harvested and restimulated on IFN-γ FluoroSpot plates with peptide-loaded monocytes. Following overnight stimulation, FluoroSpot plates were developed according to the manufacturer's protocol. The number of pan-T cells secreting IFN-γ was measured using a Mabtech IRIS™ FluoroSpot plate reader. Additionally, flow cytometry was used for moDC quality control. In addition to the target peptide, appropriate negative controls (DMSO or a control pool of 29 peptides derived from myelin oligodendrocyte glycoproteins (MOGs)) and positive controls (MART, CEF / CEFBA) were used. Materials required sterility, with endotoxin levels below 0.1 EU / mg and protein concentrations ≥1 mg / ml.

[0814] For analysis, IFNγ+ spot-forming units (SFU) for each R1 and R2 were measured using the Fluorospot assay, and δSFU (dSFU) was calculated as the absolute difference in spots between the tested smORF-derived peptide and the corresponding DMSO control conditions after normalizing the SFU per well to per million SFU.

[0815] In Examples 6.1 and 6.2 (combined peptides), a response is considered a "recall response" if the following criteria are met: i) the dSFU of R1 is at least 30, and ii) the dSFU of R2 is at least twice as high as the dSFU of R1.

[0816] In Example 6.3 (single peptide), a response is considered a "recall response" if the following criteria are met:

[0817] i) The dSFU of R1 is at least 5, and ii) The dSFU of R2 is at least twice as high as the dSFU of R1.

[0818] A response is considered a "memory response" if the following criteria are met:

[0819] i) The dSFU of R1 is at least 100, and ii) The dSFU of R2 is 0.1 to 2.0 times that of R1.

[0820] It should be noted that failure to meet the above criteria under given conditions does not necessarily mean the complete absence of recall or memory responses. Rather, it may mean that the intensity of the recall or memory response is below the above threshold, or that it is incompatible with the criteria selected for uniform batch analysis.

[0821] Example 6.1: Antigen Validation - In Vitro Immunogenicity of Melanoma Cancer Antigen - Combined Peptides

[0822] The purpose of this experiment was to analyze the in vitro immunogenicity of different melanoma cancer antigens derived from tumor-specific neoantigens and encoded by smORFs derived from lncRNA.

[0823] In this assay, cryopreserved PBMCs from 10 (healthy) donors with a range of different HLA types were tested using five different peptide pools derived from melanoma cancer antigens (see Table E4) (see Table E3).

[0824] Table E3: Overview of PBMC donors and corresponding HLA alleles used in Example 6.1

[0825]

[0826] Table E4: Overview of the melanoma peptide pool used in Example 6.1

[0827]

[0828] result:

[0829] The in vitro immunogenicity of the melanoma cancer antigen using the peptide pool described in Table E4 is as follows: Figures 10 to 14 As shown in the figure. The graph displays the IFN-γ readings measured as described in this embodiment. A summary of recall and memory responses, defined according to the above-described uniform batch analysis standards, is presented in... Figure 15 As shown in Table E5.

[0830] Table E5: Figure 15 The summary of recall and memory responses shown in the figure

[0831]

[0832] like Figure 10 As shown, seven PBMC donors exhibited antigen-specific T cell responses to at least one peptide in pool 1. One PBMC donor exhibited a memory response to at least one peptide in pool 1. (See also...) Figure 15 / Table E5). For example... Figure 11 As shown, seven PBMC donors exhibited antigen-specific T cell responses to at least one peptide in pool 2. Two PBMC donors exhibited memory responses to at least one peptide in pool 2 (see also...). Figure 15 / Table E5). For example... Figure 12 As shown, 10 PBMC donors exhibited antigen-specific T cell responses to at least one peptide in pool 3. Four PBMC donors exhibited recall responses to at least one peptide in pool 3. Four PBMC donors also exhibited memory responses to at least one peptide in pool 3 (see also...). Figure 15 / Table E5). For example... Figure 13 As shown, 8 PBMC donors exhibited antigen-specific T cell responses to at least one peptide in pool 4. 2 PBMC donors exhibited recall responses to at least one peptide in pool 4. 1 PBMC donor exhibited a memory response to at least one peptide in pool 4 (see also...). Figure 15 / Table E5). For example... Figure 14 As shown, four PBMC donors exhibited antigen-specific T cell responses to at least one peptide in pool 5. One PBMC donor exhibited a memory response to at least one peptide in pool 5. Two PBMC donors exhibited a memory response to at least one peptide in pool 5 (see also...). Figure 15 / Table E5).

[0833] In summary, each peptide pool contains at least one cancer antigen that can elicit an immune response from human T cells. Both recall and memory responses were observed in this experimental setup.

[0834] Example 6.2: Antigen Validation - In Vitro Immunogenicity of NSCLC and Melanoma Cancer Antigens - Combined Peptides

[0835] The purpose of this experiment was to analyze the in vitro immunogenicity of different NSCLC and melanoma cancer antigens derived from tumor-specific neoantigens and encoded by lncRNA-derived smORFs.

[0836] In this assay, cryopreserved PBMCs from 10 (healthy) donors with a range of different HLA types were tested (see Table E6), as well as eight different peptide pools derived from NSCLC antigen and melanoma cancer antigen (see Table E7).

[0837] Table E6: Overview of PBMC donors and corresponding HLA alleles used in Example 6.2

[0838]

[0839] Table E7: Overview of the NSCLC / melanoma peptide pool used in Example 6.2

[0840]

[0841] result:

[0842] Exemplary in vitro immunogenicity results of the NSCLC / melanoma cancer antigen and peptide pool described in Table E7 are as follows: Figure 15 As shown in the figure. This figure illustrates the IFN-γ readings measured as described in this embodiment. Figure 15 The recall and memory responses shown are further summarized in Table E8.

[0843] Table E8: Figure 16 The summary of recall and memory responses shown in the figure

[0844]

[0845] The condition pool 8 / donor D9 in Example 6.2 exhibited an immune response, characterized by dSFU R1 and dSFU R2 of 21.25 and 236, respectively.

[0846] In summary, the lncRNA-encoded peptides (lncCLEC2D91412616, KCNMB2AS14633753, lncZC3H86163144, lncNTF35511951417, and lncWDR722415641696) exhibit immunogenicity. Notably, the observed in vitro immunogenicity of these lncRNA-encoded peptides is comparable to or even superior to that of the well-known immunogenic TAA MART-1 (lncNTF35511951417 and lncWDR722415641696), further highlighting their potential as cancer targets.

[0847] Example 6.3: Antigen Validation - In Vitro Immunogenicity of Melanoma Cancer Antigen - Single Peptide

[0848] The purpose of this experiment was to analyze the in vitro immunogenicity of different melanoma cancer antigens derived from tumor-specific neoantigens and encoded by lncRNAs.

[0849] Cryopreserved PBMCs from five (healthy) donors with a range of different HLA types were tested (see Table E9) as well as nine single peptides derived from melanoma cancer antigens (see Table E10).

[0850] Table E9: Overview of PBMC donors and corresponding HLA alleles used in Example 6.3

[0851]

[0852] Table E10: Overview of a single peptide in melanoma

[0853]

[0854] In future experiments, more single peptides from pools 1 and 2 in Table E7 will be tested.

[0855] result:

[0856] Exemplary in vitro immunogenicity results using the single peptide melanoma cancer antigen described in Table E10 are as follows: Figure 17 As shown in the figure. This figure illustrates the IFN-γ readings measured as described in this embodiment. Figure 17 The recall responses shown are further summarized in Table E10A.

[0857] Table E10A: Figure 17 The summary of recall responses shown in the figure

[0858]

[0859] Example 6.4: Antigen Validation - In Vitro Immunogenicity of NSCLC and Melanoma Cancer Antigens - Single Peptide

[0860] The aim of this experiment was to analyze the in vitro immunogenicity of different NSCLC cancer antigens derived from tumor-specific neoantigens and encoded by lncRNAs. In this assay, single NSCLC peptides selected from Table E7 were used to test cryopreserved PBMCs from multiple (healthy) donors with a range of different HLA types.

[0861] Example 7: Antigen Validation - Immunopeptidomics

[0862] To further investigate and confirm epitope presentation on MHC-I / II tumor cells, immunopeptidomics analysis was performed. For this purpose, non-targeted mass spectrometry analysis was performed on primary samples from 15 melanoma (including BRAF mutations) and 15 NSCLC (10 LUSC + 5 LUAD) patients, following standard procedures in the field. In addition, whole-genome sequencing, RNA-seq, and Ribo-seq were performed on the corresponding samples.

[0863] Example 8: Preparation of nucleic acids encoding cancer antigens

[0864] This embodiment provides a method for obtaining the RNA of the present invention, and a method for generating the composition of the present invention comprising nucleic acids, particularly RNA, formulated in a lipid-based carrier.

[0865] Example 8.1. Preparation of DNA template for in vitro RNA transcription

[0866] DNA sequences encoding the cancer antigens of this invention were prepared and used in subsequent in vitro RNA transcription reactions. Several DNA sequences were prepared by modifying wild-type or reference coding DNA sequences with G / C-optimized coding sequences for stabilization and expression optimization. The sequences were introduced into pUC-derived DNA vectors to contain a stabilized UTR sequence, an adenosine segment, and optionally a histone stem-loop (hSL). The obtained plasmid DNA template was transformed and propagated in bacteria using commonly known protocols in the art. Finally, the plasmid DNA template was extracted, purified, and linearized using type II restriction endonucleases. The RNA constructs used herein are provided in Table E11.

[0867] Table E11: RNA constructs used in the examples (smORF nomenclature is shown in Table E12)

[0868]

[0869] Table E12: smORF nomenclature used in Table E11

[0870]

[0871] Example 8.2. In vitro transcription of RNA from a plasmid DNA template

[0872] Under appropriate buffer conditions, linearized DNA templates were used for DNA-dependent RNA in vitro transcription (IVT) using T7 RNA polymerase in the presence of a sequence-optimized nucleotide mixture (ATP / GTP / CTP / UTP) and a cap analogue (for Cap1: m7G(5')ppp(5')(2'OMeA)pG; TriLink). Other constructs were produced in the presence of a nucleotide mixture containing (ATP / GTP / CTP / pseudouridine(ψ)) or (ATP / GTP / CTP / N1-methylpseudouridine(m1ψ)) and a cap analogue. Following RNA in vitro transcription, the resulting RNA IVT reaction was purified using steps including RP-HPLC.

[0873] Example 8.3 Preparation of a lipid-based vector for encapsulating mRNA

[0874] The LNPs used in the working examples were fabricated using the NanoAssemblr™ microfluidic system (Precision NanoSystems Inc., Vancouver, BC) according to a standard protocol. This system enables controlled, bottom-up molecular self-assembly of nanoparticles through a custom-designed microfluidic mixing chip, which allows for millisecond-level mixing of nanoparticle components at the nanoliter scale. To prepare the lipid nanoparticle compositions, the following excipients / lipids were used: (i) ionizable lipids: VitE-C4DE-Pip-thioether as described herein and in Table 1 of published patent application WO2021123332 (compound C26); (ii) cholesterol as described herein (Avanti Polar Lipids; Alabaster, AL); (iii) neutral lipid / phospholipid “DPhyPE” (1,2-diphydanyl-sn-glycerol-3-phosphate ethanolamine; Avanti Polar Lipids; Alabaster, AL) as described herein; (iv) phosphatidylserine DPhyPS (1,2-diphydanyl-sn-glycerol-3-phosphate-L-serine; Avanti Polar Lipids; Alabaster, AL, 850408P) as described herein; and (v) polymer-conjugated lipids as described herein, PMOZ-lipid “PMOZ 4”. Further characterization of the LNP compositions is described in Table E13.

[0875] Table E13: LNP Compositions

[0876]

[0877] The lipids were dissolved in an alcoholic solution (ethanol) according to standard procedures. Specifically, LNPs were prepared by mixing an appropriate volume of a lipid stock solution of cholesterol, phospholipids, and polymer-conjugated lipids in an ethanol buffer (20 mg / ml, dissolved in EtOH); cationic lipid C26 was dissolved in 30 mg / ml tert-butanol and added to the ethanol premix of lipids. This ethanol mixture of lipids was then combined with an aqueous phase (50 mM sodium acetate, pH 4.0) containing 1 g / L mRNA. Briefly, the mRNA indicated in the working example was diluted to 0.05 mg / ml to 0.2 mg / ml in 50 mM acetate buffer (pH 4). A syringe pump was attached to the inlet unit of a NanoAssemblr™ (Precision NanoSystems Inc., Vancouver, BC) to mix the ethanol lipid solution and the aqueous mRNA solution at a ratio of approximately 1:5 to 1:3 (volume / volume), with a total flow rate of approximately 14 ml / min to approximately 18 ml / min. Ethanol was then removed by dialysis (Slide-A-Lyzer™ Dialysis Cassettes, ThermoFisher), and the external buffer was replaced with PBS / sucrose buffer (pH 7.4, 75 mM NaCl, 10 mM phosphate, 150 mM sucrose). Finally, the lipid nanoparticles were filtered through a sterile filter with a 0.2 μm pore size. The particle diameter of the lipid nanoparticles was determined to be approximately 90 nm to approximately 140 nm using quasi-elastic light scattering on a Malvern Zetasizer Nano (Malvern Instruments Ltd.; UK).

[0878] Example 9: Translation of smORF-encoded mRNA and stability of the translated protein

[0879] According to standard procedures known in the art, the translation of smORF-encoded mRNAs (mRNA constructs described in Table E11) and the stability of expressed proteins were measured using Western blotting and flow cytometry. HEK293T cells were transfected with 1 µg / ml Lipofectamin (400,000 cells / well, 200,000 cells / ml), and the amount of expressed protein was measured by FACS using antibodies against the CTLA4 domain or HA-tag after 4 h and 16 to 18 h of incubation with or without carfilzomib. Additionally, HeLa cells were transfected with mRNA, treated for 2 h with or without carfilzomib, and harvested and lysed after 18 h of treatment. Subsequently, the amount of expressed protein was assessed by Western blotting using antibodies against the HA-tag. The translation of mRNA constructs encoding single or multiple smORFs and the stability of their encoded fusion proteins were confirmed by CTLA-4 or HA staining using FACS and Western blotting.

[0880] Example 10: In vitro immunogenicity of mRNA constructs encoding multiple smORFs and / or TSAs

[0881] mRNA constructs encoding multiple smORFs and / or TSAs are produced as previously described (see R12300 and R12301 in Example 8). Alternatively, variants with auxiliary epitopes replacing the HA tag are produced as follows:

[0882] Variant R12300 (encoding mRNAs of two smORFs expressed in melanoma): 5'UTR-SP(CTLA4)-adaptor(GS4)-smORF1-adaptor(GS4)-smORF2-adaptor(GS4)-helper epitope-adaptor(GS4)-TMD / TMCD(CTLA4)-3'UTR-PolyA.

[0883] Variant R12301 (encoding mRNAs of six smORFs expressed in NSCLC): 5'UTR-SP(CTLA4)-adaptor(GS4)-smORF2-adaptor(GS4)-smORF3-adaptor(GS4)-smORF5-adaptor(GS4)-smORF6-adaptor(GS4)-smORF7-adaptor(GS4)-smORF4-adaptor(GS4)-auxiliary epitope-adaptor(GS4)-TMD / TMCD(CTLA4)-3'UTR-PolyA.

[0884] The in vitro immunogenicity of the mRNA construct was analyzed using assays similar to those described in Example 6. The first stimulation cycle was performed using mRNA, and the second stimulation cycle was performed using the corresponding peptide.

[0885] Example 11: In vivo immunogenicity of the mRNA construct encoding smORF

[0886] To analyze the in vivo immunogenicity of the mRNA construct encoding smORF, the R12300 and R12301 constructs were produced and formulated as described in Example 8, and tested as follows.

[0887] To enable immunization, the construct was administered intramuscularly (im; tibialis muscle) to female CB6F1 hybrid mice at doses of 5 µg LNP-prepared mRNA (R12300, R12301 or PpLuc control mRNA R8730) on days 0, 7 and 14.

[0888] Twenty-one days after the first mRNA administration, mice were sacrificed and spleens were collected for further analysis. Cancer antigen-specific cellular responses in the spleen cell samples obtained in this step were measured as antigen-specific T cell activation. This was performed according to a standard protocol using intracellular cytokine staining and subsequent flow cytometry analysis: isolated spleen cells were individually restimulated for 6 hours in the presence of anti-CD28 (BD Biosciences, San Jose, USA) with a peptide library targeting each smORF (a 15-mer library covering the complete antigen) at a final peptide concentration of 1 µg / ml, followed by additional GolgiPlug (BD Biosciences) after 1 hour. Unstimulated spleen cells were treated in the same manner but supplemented with DMSO instead of the peptide mixture. Other controls included spleen cells stimulated with PMA / iomycin (without anti-CD28; PMA and iomycin were from Sigma-Aldrich; MerckKGaA, Darmstadt, Germany) (positive control) and spleen cells stained with unconjugated antibodies (negative control). Following stimulation, spleen cells were stained with antibodies conjugated to surface and intracellular fluorophores and analyzed by flow cytometry. CD8 and CD4 responses were indicated by IFN-γ / TNF-α double-positive CD8 T cells and CD4 T cells.

[0889] result:

[0890] Example data for the R12300 constructor is as follows Figure 18 As shown. Data shows that mRNA R12300 elicited a potent CD8 response (top) and CD4 response (bottom) against the lncCLEC2D91412616 smORF. Exemplary data for the construct R12301 and T cell responses against NKZW-smORF (KCNMB2AS14633753, lncNTF35511951417, lncWDR722415641696, lncZC3H86163144) are shown below. Figure 19 As shown, the data indicate that R12301 elicited effective CD8 responses (top) and CD4 responses (bottom). In summary, these data highlight that mRNAs encoding smORFs can elicit effective immune responses against these encoded targets in vivo.

[0891] Example 12: Broad-based lncRNA indication screening

[0892] The purpose of this experiment was to analyze whether lncRNAs KCNMB2-AS1...

Claims

1. An artificial nucleic acid comprising at least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising or constituting at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA), or immunogenic fragments or variants thereof.

2. The artificial nucleic acid according to claim 1, wherein the at least one antigenic peptide is selected from peptides or proteins encoded by long non-coding RNA (lncRNA), or any immunogenic fragments or variants thereof, wherein the long non-coding RNA is selected from lnc-NTF3-5:5, lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-ZC3H8-6:1, lnc-CLEC2D-9:1, lnc-TRPC5-3:1, or LINC00893:

25.

3. The artificial nucleic acid according to claims 1 to 2, wherein at least one antigenic peptide encoded by a long non-coding RNA (lncRNA) contains at least one T-cell epitope, preferably at least one CD8+ T-cell epitope.

4. The artificial nucleic acid according to claims 1 to 3, wherein at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA) comprises or constitutes at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these.

5. The artificial nucleic acid according to claims 1 to 4, comprising: • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from lnc-NTF3-5:5 encoded by a peptide or protein, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these; or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-WDR72-2:4, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these; or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from lnc-TRPC5-3:1 encoding a peptide or protein, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these; or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising: at least one antigenic peptide selected from the peptide or protein encoded by lnc-CLEC2D-9:1, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these; or an immunogenic fragment or variant thereof; or At least one antigenic peptide selected from the peptide or protein encoded by KCNMB2-AS1:4, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence that is identical to or has at least 80% identity with any one of SEQ ID NO: 147 to SEQ ID NO: 161, SEQ ID NO: 572 to SEQ ID NO: 589, or SEQ ID NO: 758, or any of these immunogenic fragments or variants; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-ZC3H8-6:1, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these; or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by LINC00893:25, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these, or an immunogenic fragment or variant thereof.

6. The artificial nucleic acid according to claims 1 to 5, comprising: • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-NTF3-5:5, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 80% identity with SEQ ID NO:760, or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-WDR72-2:4, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 80% identity with SEQ ID NO:756, or an immunogenic fragment or variant thereof; or • At least one antigenic peptide selected from the peptide or protein encoded by KCNMB2-AS1:4, wherein the antigenic peptide comprises or constitutes at least one amino acid sequence identical to or having at least 80% identity with SEQ ID NO: 758, or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-ZC3H8-6:1, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 80% identity with SEQ ID NO:759, or an immunogenic fragment or variant thereof; or • At least one coding sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-CLEC2D-9:1, said antigenic peptide comprising or constituting at least one amino acid sequence identical to or having at least 80% identity with SEQ ID NO: 762, or an immunogenic fragment or variant thereof.

7. The artificial nucleic acid according to claims 1 to 6, wherein the at least one coding sequence comprises a nucleic acid sequence, preferably a GC-optimized nucleic acid sequence, the nucleic acid sequence encoding at least one antigenic peptide selected from lncRNA-encoded peptides or proteins, wherein the nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NO: 318 to SEQ ID NO: 448, SEQ ID NO: 651 to SEQ ID NO: 755, SEQ ID NO: 764 to SEQ ID NO: 771, or any fragments or variants thereof.

8. The artificial nucleic acid according to claims 1 to 7, comprising: • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-NTF3-5:5, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants; or • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-WDR72-2:4, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants; or • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-TRPC5-3:1, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants; or • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-CLEC2D-9:1, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants; or • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by KCNMB2-AS1:4, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants; or • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-ZC3H8-6:1, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants; or • At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, said tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by LINC00893:25, wherein said nucleic acid sequence is identical to or has at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of SEQ ID NO: 438 to SEQ ID NO: 448, SEQ ID NO: 771, or any of these fragments or variants.

9. The artificial nucleic acid according to any one of the preceding claims, wherein the at least one coding sequence encodes 2 to 10 different tumor antigens, preferably 2 to 5 different tumor antigens.

10. The artificial nucleic acid according to any one of the preceding claims, wherein the at least one coding sequence encodes at least one additional amino acid sequence, said additional amino acid sequence being selected from at least one immune response activation signal transduction protein, at least one T helper cell epitope, and / or at least one signal peptide.

11. The artificial nucleic acid according to any one of the preceding claims, wherein the at least one coding sequence is a codon-modified coding sequence, preferably wherein the codon-modified coding sequence is selected from a C-maximized coding sequence, a CAI-maximized coding sequence, a human codon selection-adapted coding sequence, a G / C content-modified coding sequence, and a G / C-optimized coding sequence, or any combination thereof.

12. The artificial nucleic acid according to claim 11, wherein the at least one codon-modified coding sequence is a G / C optimized coding sequence.

13. The artificial nucleic acid according to any one of the preceding claims, wherein the nucleic acid comprises at least one untranslated region (UTR), the untranslated region preferably selected from at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR.

14. The artificial nucleic acid of claim 13, wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence from a 3'-UTR selected from, or from a homolog, fragment, or variant of any of these genes: PSMB3, ALB7, α-globin, β-globin, ANXA4, CASP1, COX6B1, FIG4, GNAS, NDUFA1, RPS9, SLC7A3, or TUBB4B, preferably wherein the at least one heterologous 3'-UTR comprises or constitutes a nucleic acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these fragments or variants.

15. The artificial nucleic acid of claim 13, wherein the at least one heterologous 5'-UTR comprises or is composed of a nucleic acid sequence derived from a 5'-UTR of a gene selected from, or from a homolog, fragment, or variant of any of these genes: HSD17B4, RPL32, AIG1, α-globin, ASAH1, ATP5A1, COX6C, DPYSL2, MDR, MP68, NDUFA4, NOSIP, RPL31, RPL35A, SLC7A3, TUBB4B, or UBQLN2, preferably wherein the at least one heterologous 5'-UTR comprises or is composed of a sequence from SEQ ID NO: 12 to SEQ ID NO: 45, SEQ ID NO: 64, SEQ ID NO:

65. Or any of these fragments or variants identical or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity nucleic acid sequences.

16. The artificial nucleic acid according to claims 13 to 15, wherein the at least one heterologous 5'-UTR is selected from HSD17B4 and the at least one heterologous 3' UTR is selected from PSMB3.

17. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid is selected from DNA or RNA, preferably RNA.

18. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid is an RNA selected from mRNA, circular RNA, replicon RNA, or viral RNA, preferably mRNA.

19. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid, preferably the RNA, comprises at least one poly(A) sequence, preferably wherein the at least one poly(A) sequence comprises about 40 to about 500 adenosine nucleotides.

20. The artificial nucleic acid according to claim 19, wherein the at least one poly(A) sequence comprises about 60 to about 150 adenosine nucleotides, preferably about 100 adenosine nucleotides.

21. The artificial nucleic acid according to claim 19 or 20, wherein the at least one poly(A) sequence is located at the 3' end, optionally wherein the 3' terminal nucleotide is adenosine.

22. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid, preferably the RNA, comprises at least one histone stem-loop sequence, wherein the histone stem-loop sequence preferably comprises or constitutes a nucleic acid sequence that is identical to or has at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of the fragments or variants of SEQ ID NO: 3 or SEQ ID NO:

4.

23. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid is RNA, the RNA comprising at least one modified nucleotide, preferably a modified nucleotide selected from pseudouridine (ψ) or N1-methylpseudouridine (m1ψ), more preferably N1-methylpseudouridine (m1ψ).

24. The artificial nucleic acid according to claims 1 to 22, wherein the nucleic acid is RNA, and the RNA does not contain modified nucleotides.

25. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid is RNA, and the RNA comprises a 5'-cap structure, preferably a cap1 structure or a modified cap1 structure.

26. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid is RNA transcribed in vitro.

27. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid, preferably the RNA, comprises the following sequence elements, preferably in the 5' to 3' orientation: A) 5'-cap structure; B) 5'-UTR, preferably selected from the 5'-UTR of the HSD17B4 gene; C) A coding sequence that encodes at least one tumor antigen; D) 3'-UTR, preferably selected from the 3'-UTR of the PSMB3 gene; E) Optionally, histone stem-loop; and F) poly(A) sequence, which preferably contains about 100 A nucleotides.

28. The artificial nucleic acid according to any one of the preceding claims, wherein the artificial nucleic acid is mRNA, and the mRNA comprises the following sequence elements in the 5' to 3' orientation: A) 5'-cap structure; B) 5'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 80% identity with SEQ ID NO: 13, or a fragment or variant thereof; C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from long non-coding RNA (lncRNA) encoded by at least one antigenic peptide or protein, wherein the nucleic acid sequence is identical or has at least 80% identity with any one of SEQ ID NO: 318 to SEQ ID NO: 448, SEQ ID NO: 651 to SEQ ID NO: 755, SEQ ID NO: 764 to SEQ ID NO: 771, or any fragment or variant thereof; D) 3'-UTR, which contains or constitutes a nucleic acid sequence that is identical to or has at least 80% identity with SEQ ID NO: 67, or a fragment or variant thereof; E) Optionally, histone stem-loop; and F) poly(A) sequence, which preferably contains about 100 A nucleotides.

29. A pharmaceutical composition comprising at least one artificial nucleic acid, said artificial nucleic acid comprising at least one coding sequence encoding at least one tumor antigen as defined in any one of claims 1 to 28.

30. The pharmaceutical composition of claim 29, comprising a plurality of artificial nucleic acids, each of which encodes at least one different tumor antigen.

31. The pharmaceutical composition according to claim 29 or 30, wherein the at least one artificial nucleic acid, preferably the RNA, is formulated in at least one cationic compound or polycationic compound, wherein the at least one cationic compound or polycationic compound is optionally selected from cationic polymers or polycationic polymers, cationic polysaccharides or polycationic polysaccharides, cationic lipids or polycationic lipids, cationic proteins or polycationic proteins, cationic peptides or polycationic peptides, or any combination thereof.

32. The pharmaceutical composition according to claims 29 to 31, wherein the at least one artificial nucleic acid, preferably the RNA, is formulated in a lipid-based carrier.

33. The pharmaceutical composition of claim 32, wherein the lipid-based carrier is selected from liposomes, lipid nanoparticles, lipid complexes, solid lipid nanoparticles, lipid-polymer complexes, and / or nanoliposomes.

34. The pharmaceutical composition according to claim 32 or 33, wherein the lipid-based carrier is a lipid nanoparticle.

35. The pharmaceutical composition according to claims 32 to 34, wherein the lipid-based carrier, preferably the LNP, comprises... (i) at least one cationic lipid; (ii) at least one neutral lipid; (iii) at least one steroid or steroid analogue; and (iv) At least one lipid that reduces aggregation.

36. The pharmaceutical composition according to claims 32 to 37, wherein the lipid-based carrier has a Z-average particle size of about 50 nm to about 200 nm, preferably about 50 nm to about 150 nm.

37. The pharmaceutical composition or artificial nucleic acid according to any one of the preceding claims, wherein when the composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to a cell, tissue, or object, an encoded tumor antigen is generated.

38. The pharmaceutical composition or artificial nucleic acid according to any one of the preceding claims, wherein when the composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or objects, it generates an encoded tumor antigen and induces epitope-specific CD8+ T cells in the object.

39. A composition of a tumor antigen or at least one tumor antigen, wherein the tumor antigen is characterized by any one of the preceding claims, preferably by any one of claims 2 to 6, and more preferably wherein the tumor antigen comprises or constitutes at least one amino acid sequence identical to or having at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any of these.

40. An antibody, T cell, or TCR that is generated or stimulated with any of the tumor antigens or nucleic acids as defined in any of the preceding claims, preferably generated or stimulated with the tumor antigen as defined in claim 39.

41. A combination comprising at least two or more treatment modalities, said treatment modalities being selected from... a) at least one artificial nucleic acid as defined in claims 1 to 28; and / or b) At least one pharmaceutical composition as defined in claims 29 to 38; and / or c) At least one tumor antigen or composition as defined in claim 39; and / or d) At least one antibody, T cell, or TCR as defined in claim 40.

42. A kit or kit comprising at least one artificial nucleic acid as claimed in any one of claims 1 to 28, and / or at least one pharmaceutical composition as claimed in any one of claims 29 to 38, and / or at least one tumor antigen or composition of tumor antigen as claimed in claim 39, and / or at least one antibody, T cell or TCR as claimed in claim 40, optionally comprising a liquid carrier for dissolution, and optionally comprising technical instructions providing information on the administration and dosage of the components.

43. An artificial nucleic acid as described in any one of claims 1 to 28, a pharmaceutical composition as described in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as described in claim 39, an antibody, T cell or TCR as described in claim 40, a combination as described in claim 41, or a kit or kit as described in claim 42, used as a pharmaceutical agent.

44. An artificial nucleic acid as described in any one of claims 1 to 28, a pharmaceutical composition as described in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as described in claim 39, an antibody, T cell or TCR as described in claim 40, a combination as described in claim 41, or a kit or kit as described in claim 42, used as a medicine for treating or preventing cancer or any cancer-related disease, disorder or condition in a subject.

45. An artificial nucleic acid as described in any one of claims 1 to 28, a pharmaceutical composition as described in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as described in claim 39, an antibody, T cell or TCR as described in claim 40, a combination as described in claim 41, or a kit or kit as described in claim 42, used as a medicine for treating or preventing squamous cell carcinoma or any disease, disorder or condition associated with squamous cell carcinoma in a subject.

46. ​​An artificial nucleic acid as described in any one of claims 1 to 28, a pharmaceutical composition as described in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as described in claim 39, an antibody, T cell or TCR as described in claim 40, a combination as described in claim 41, or a kit or kit as described in claim 42, used as a medicament for treating or preventing NSCLC or any disease, disorder or condition associated with NSCLC, particularly squamous cell carcinoma / squamous non-small cell lung cancer, in subjects.

47. Use as a medicament for the treatment or prevention of NSCLC according to claim 46, wherein at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises at least one antigenic peptide, the antigenic peptide being selected from peptides or proteins encoded by long non-coding RNA (lncRNA), or any fragments or variants thereof, the long non-coding RNA being selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:

1.

48. An artificial nucleic acid as claimed in any one of claims 1 to 28, a pharmaceutical composition as claimed in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as claimed in claim 39, an antibody, T cell or TCR as claimed in claim 40, a combination as claimed in claim 41, or a kit or kit as claimed in claim 42, used as a medicine for treating or preventing HNSCC or any disease, disorder or condition associated with HNSCC in a subject.

49. Use as a medicament for the treatment or prevention of HNSCC according to claim 48, wherein at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises at least one antigenic peptide, the antigenic peptide being selected from peptides or proteins encoded by long non-coding RNA (lncRNA), or any fragments or variants thereof, the long non-coding RNA being selected from lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:

1.

50. An artificial nucleic acid as claimed in any one of claims 1 to 28, a pharmaceutical composition as claimed in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as claimed in claim 39, an antibody, T cell or TCR as claimed in claim 40, a combination as claimed in claim 41, or a kit or kit as claimed in claim 42, used as a medicine for treating or preventing melanoma or any disease, disorder or condition associated with melanoma in a subject.

51. Use as a medicament for treating or preventing melanoma according to claim 50, wherein at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises at least one antigenic peptide, the antigenic peptide being selected from peptides or proteins encoded by long non-coding RNA (lncRNA), or any fragments or variants thereof, the long non-coding RNA being selected from lnc-CLEC2D-9:1, lnc-TRPC5-3:1, or LINC00893:

25.

52. An artificial nucleic acid as described in any one of claims 1 to 28, a pharmaceutical composition as described in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as described in claim 39, an antibody, T cell or TCR as described in claim 40, a combination as described in claim 41, or a kit or kit as described in claim 42, for use in treating or preventing adrenocortical carcinoma, bladder urothelial carcinoma, invasive breast cancer, cervical cancer (cervical squamous cell carcinoma and cervical adenocarcinoma), colonic adenocarcinoma, renal papillary carcinoma, etc. A drug for cancer, lung adenocarcinoma, lymphoid tumor, diffuse large B-cell lymphoma, rectal adenocarcinoma, sarcoma, skin melanoma, thymoma, thyroid cancer, hepatocellular carcinoma, uterine carcinosarcoma, or endometrial cancer, or any related disease, disorder, or condition, wherein at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or a composition of tumor antigens, the combination, or the kit or kit kit comprises at least one antigenic peptide, the antigenic peptide being selected from peptides or proteins encoded by lnc-ZC3H8-6:1, or fragments or variants thereof.

53. A medicament for use in treating or preventing bladder urothelial carcinoma, cervical cancer (squamous cell carcinoma and adenocarcinoma of the cervix), thymoma, or endometrial cancer, or any related disease, disorder, or condition in a subject, wherein at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or the composition of the tumor antigen, the combination, or the kit or kit comprises at least one antigenic peptide selected from peptides or proteins encoded by KCNMB2-AS1:4, or fragments or variants thereof.

54. An artificial nucleic acid as claimed in any one of claims 1 to 28, a pharmaceutical composition as claimed in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as claimed in claim 39, an antibody, T cell, or TCR as claimed in claim 40, a combination as claimed in claim 41, or a kit or kit as claimed in claim 42, used as a medicament for treating or preventing ovarian serous cystadenocarcinoma, testicular germ cell tumor, thymoma, or uterine carcinosarcoma, or any related disease, disorder, or condition in a subject, wherein at least one tumor antigen provided by said artificial nucleic acid, said pharmaceutical composition, said tumor antigen or a composition of tumor antigens, said combination, or said kit or kit comprises at least one antigenic peptide, said antigenic peptide being selected from a peptide or protein encoded by lnc-NTF3-5:5, or a fragment or variant thereof.

55. An artificial nucleic acid as claimed in any one of claims 1 to 28, a pharmaceutical composition as claimed in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as claimed in claim 39, an antibody, T cell, or TCR as claimed in claim 40, a combination as claimed in claim 41, or a kit or kit as claimed in claim 42, used as a medicament for treating or preventing bladder urothelial carcinoma, colon adenocarcinoma, esophageal cancer, or gastric adenocarcinoma, or any related disease, disorder, or condition in a subject, wherein at least one tumor antigen provided by said artificial nucleic acid, said pharmaceutical composition, said tumor antigen or a composition of tumor antigens, said combination, or said kit or kit comprises at least one antigenic peptide, said antigenic peptide being selected from peptides or proteins encoded by lnc-WDR72-2:4, or fragments or variants thereof.

56. An artificial nucleic acid as claimed in any one of claims 1 to 28, a pharmaceutical composition as claimed in any one of claims 29 to 38, a tumor antigen or a composition of tumor antigens as claimed in claim 39, an antibody, T cell, or TCR as claimed in claim 40, a combination as claimed in claim 41, or a kit or kit as claimed in claim 42, used as a medicament for treating or preventing cutaneous melanoma, testicular germ cell tumors, or uveal melanoma, or any related disease, disorder, or condition in a subject, wherein at least one tumor antigen provided by said artificial nucleic acid, said pharmaceutical composition, said tumor antigen or a composition of tumor antigens, said combination, or said kit or kit comprises at least one antigenic peptide, said antigenic peptide being selected from peptides or proteins encoded by lnc-CLEC2D-9:1, or fragments or variants thereof.

57. A method for treating or preventing a disease, disorder, or ailment, wherein the method comprises applying or administering to a subject in need an effective amount of an artificial nucleic acid as claimed in claims 1 to 28, a composition as claimed in claims 29 to 38, a tumor antigen or a composition of tumor antigens as claimed in claim 39, an antibody, T cell, or TCR as claimed in claim 40, a combination as claimed in claim 41, or a kit or kit as claimed in claim 42.

58. The method of claim 57, wherein the method is further characterized by the medical use features as defined in claims 43 to 56.

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