Cancer antigens

Abstract

The present invention relates to tumor antigens or cancer antigens, particularly amino acid sequences and nucleic acid sequences, which may be used in cancer immunotherapy. In particular, the present invention relates to artificial nucleic acids, preferably RNA, comprising at least one coding sequence encoding at least one tumor antigen or cancer antigen. The tumor antigen or cancer antigen is Peptides or proteins encoded by small open reading frames (smORFs) of long non-coding RNA (IncRNA), or The fragment or variant thereof, At least one antigenic peptide selected from or derived therefrom, and / or Tumor neoantigen peptides or proteins, or The fragment or variant thereof, at least one antigenic peptide selected from or derived therefrom The invention provides a pharmaceutical composition comprising or consisting of the artificial nucleic acid, preferably formulated within a lipid-based carrier. It also provides a method for treating or preventing a disease, disorder, or condition, and medical applications for treating or preventing cancer in particular (such as NSCLC, HNSCC, or melanoma).

Description

Detailed Description of the Invention

[0001] 〔Introduction〕 Cancer is a global health problem and one of the major causes of death worldwide. Conventional cancer treatments (such as surgery, chemotherapy, and radiotherapy) have limited effectiveness and can cause significant side effects. Immunotherapy utilizes the power of the immune system targeting cancer cells and has emerged as a promising new approach in cancer treatment. Cancer immunotherapy is a method of treating cancer using the immune system. Immunotherapy utilizes the fact that there are molecules (called tumor antigens) on the surface of cancer cells that can be recognized by the immune system. Many of these molecules are proteins or other macromolecules. Promising candidates for immunotherapy with high specificity are neoantigens, antigens, or tumor-associated antigens.

[0002] In recent years, nucleic acids such as RNA have attracted attention as promising therapeutic tools in the treatment of cancer and other diseases because they can inhibit the growth or survival of cancer cells or induce the production of proteins that can be decreased by other means. The advantages of RNA utilization include transient expression and non-transforming properties. Since RNA does not need to enter the nucleus for expression and is not integrated into the host genome, the risk of carcinogenesis can be eliminated. Therefore, nucleic acids (such as RNA) can be a promising molecular class that provides information for expressing tumor antigens (such as neoantigens or tumor-associated antigens).

[0003] Therefore, an object of the present invention is to provide neoantigens or tumor-associated antigens for cancer immunotherapy, particularly neoantigens or tumor-associated antigens provided by nucleic acid constructs.

[0004] The above objectives are achieved by the following description and appended claims. In particular, the object of the present invention is achieved by providing novel tumor antigens, nucleic acids encoding said tumor antigens, and the use of said tumor antigens / nucleic acids for cancer treatment in subjects (such as NSCLC, HNSCC, or melanoma). Furthermore, the invention provides unexpected tumor antigens derived from small open reading frames of long non-coding RNAs (which, as their name suggests, have long been considered non-coding).

[0005] [Brief Description of the Invention] The present invention relates, in particular, to novel tumor or cancer antigens, especially amino acid sequences and nucleic acid sequences, which may be used in cancer immunotherapy. In particular, the present invention relates to artificial nucleic acids, preferably RNA, comprising at least one coding sequence encoding at least one tumor or cancer antigen. The antigen is, Peptides or proteins encoded by small open reading frames (smORFs) of lncRNAs, or fragments or variants thereof, At least one antigenic peptide selected from or derived therefrom; and / or a peptide or protein of a tumor neoantigen, or a fragment or variant thereof, at least one antigenic peptide selected from or derived therefrom This includes or consists of. Furthermore, pharmaceutical compositions comprising artificial nucleic acids are provided, preferably formulated within a lipid-based carrier. Also provided are methods for treating or preventing disorders, diseases, or conditions, particularly cancer (such as NSCLC, HNSCC, or melanoma), and medical uses for such treatments.

[0006] As described herein, the present invention is based, in particular, on the remarkable discovery that peptides encoded by small open reading frames (smORFs) of long non-coding RNAs (lncRNAs) can function as effective tumor-associated antigens. Furthermore, neoantigens that can function as effective tumor-specific antigens have been identified. These tumor antigens can be provided by artificial nucleic acids (e.g., mRNA) and used for the treatment of cancers (such as NSCLC, HNSCC, or melanoma).

[0007] Therefore, in a first embodiment, the present invention provides an artificial nucleic acid comprising at least one coding sequence encoding at least one tumor or cancer antigen. The antigen is, (a) at least one antigenic peptide selected from or derived from a peptide or protein encoded by a long non-coding RNA (lncRNA); and / or (b) at least one antigenic peptide selected from or derived from a tumor neoantigen peptide or protein, Includes.

[0008] In a preferred embodiment, at least one antigenic peptide is Peptides or proteins encoded by long non-coding RNAs (lncRNAs) 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 The immunogenic fragment or its variant, Selected from or derived from.

[0009] In another preferred embodiment, at least one antigenic peptide is A peptide or protein of a tumor neoantigen selected from NRAS, TYW1B, ECPAS, MAP2K1, TOMM22, GLB1, MAGE-A3, ATAD2, BRAF, EGFR, or TP53, or The immunogenic fragment or its variant, Selected from.

[0010] In a second embodiment, the present invention provides a pharmaceutical composition comprising at least one artificial nucleic acid as defined in the first embodiment. The artificial nucleic acid (preferably RNA) may be formulated within an LNP.

[0011] In a third embodiment, the present invention provides a tumor antigen or a composition antigen.

[0012] In a fourth aspect, the present invention provides antibodies, T cells, or TCRs prepared against or stimulated by any of the at least one nucleic acids encoding the tumor antigen of the present invention.

[0013] In a fifth embodiment, the present invention provides a combination of at least two or more therapeutic modalities of any of the above embodiments.

[0014] In a sixth embodiment, the present invention is At least one artificial nucleic acid of the first embodiment; at least one pharmaceutical composition of the second embodiment; at least one tumor antigen or composition of the third embodiment; and / or at least one antibody, T cell, or TCR of the fourth embodiment. We provide a kit or partial kit that includes the above.

[0015] In a further embodiment, the present invention provides methods for treating or preventing a disease, disorder, or condition, and medical applications of artificial nucleic acids, pharmaceutical compositions, tumor antigens, or compositions, antibodies, T cells, or TCRs, or kits or component kits. Preferably, the disease, disorder, or condition is cancer, or any cancer-related disease, disorder, or condition (such as NSCLC, HNSCC, or melanoma).

[0016] [Definition] For clarity and readability, the following definitions are provided. The technical features referred to in these definitions may apply to each embodiment of the invention. Additional definitions and explanations may also be provided specifically in the context of these embodiments.

[0017] In a numerical context, percentages should be understood as relative values ​​to the total number of each item. Otherwise, unless otherwise specified in the context, percentages should be understood as weight percentages (wt.-%).

[0018] "Approximately": The term "approximately" is used when the determining factors or values ​​do not need to be identical, i.e., they do not need to be 100% identical (for example, when each value or determining factor differs by a range of 1% to 10%). Preferably, "approximately" means that the determining factors or values ​​may differ by a range of ±1%, ±2%, ±3%, ±4%, ±5%, ±6%, ±7%, ±8%, ±9%, or ±10%.

[0019] "Antigenic peptide": As used herein, the terms "antigenic peptide" or "immunogenic peptide" refer to peptides derived from (antigenic or immunogenic) proteins that stimulate the body's adaptive or cellular immune system to obtain an adaptive or cellular immune response. Therefore, an antigenic / immunogenic peptide comprises at least one epitope or antigen selected from or derived from tumor antigens as defined herein.

[0020] "Cationicity," "Cationizable": The term "cationicity" means that a structure, compound, group, or atom carries a positive charge, which may be permanent or temporary depending on certain conditions, such as pH. The terms "cationicity," "cationisable," and "permanent cationicity" as used herein should be understood as defined in WO2023 / 031394 (p.12, line 32 to p.13, line 16).

[0021] "Cationic compound," "Polycationic compound": When a structure, compound, group, or atom carries multiple positive charges, it may be referred to as polycationic. The terms used herein should be understood to be as defined in WO2021 / 156267 (p.88, line 12 to p.89, line 22).

[0022] "Coding sequence," "coding region," and "cds": As used herein, "coding sequence" or its abbreviation "cds" refers to a sequence of triplicates of multiple nucleotides that can be translated into a peptide or protein. In the context of the present invention, cds may typically be a DNA or RNA sequence consisting of multiple nucleotides in multiples of 3, beginning with a start codon and preferably ending with a stop codon. In the context of the present invention, the cds encode at least one tumor antigen as defined herein.

[0023] "derived from": As used herein in the context of nucleic acids, the term "derived from" means that a nucleic acid derived from (another) nucleic acid has 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, 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 an amino acid sequence derived from (another) amino acid sequence has 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, or is identical to the amino acid sequence from which it is derived.

[0024] "Epitope": As used herein, the term "epitope" refers to T cell epitopes and B cell epitopes. T cell epitopes, or portions of antigenic peptides or proteins, may include fragments, preferably having a length of about 6 to about 20 amino acid residues or more. For example, a fragment processed and presented by an MHC class I molecule, preferably having a length of about 8 to about 10 amino acid residues, e.g., 8, 9, or 10 (or 11 or 12). Alternatively, a fragment processed and presented by an MHC class II molecule, preferably having a length of about 13 to about 20 amino acid residues or more. These fragments are usually recognized by T cells in the form of a complex consisting of a peptide fragment and an MHC molecule; that is, fragments are usually not recognized in their native form. B cell epitopes are usually fragments located on the outer surface of (native) proteins or peptide antigens, preferably having 5 to 15 amino acids, more preferably 5 to 12 amino acids, and even more preferably 6 to 9 amino acids. These epitopes may be recognized by antibodies, i.e., recognized in their native form. Such protein or peptide epitopes may further be selected from any of the variants of the protein or peptide described herein. In this context, an epitope is, A protein or peptide segment as defined herein, which is a conformational epitope or discontinuous epitope that is discontinuous in the amino acid sequence of the protein or peptide as defined herein but close in proximity in the three-dimensional structure, or It may be a continuous epitope or a linear epitope consisting of a single polypeptide chain.

[0025] "Fragment": As used herein in the context of nucleic acid sequences (e.g., RNA or DNA) or amino acid sequences, the term "fragment" usually refers to a shorter portion of a reference sequence (e.g., a nucleic acid sequence or an amino acid sequence). Thus, a fragment usually consists of the same sequence as the corresponding continuous portion within the reference sequence. Preferred sequence fragments in the context of the present invention consist of continuous components (such as nucleotides or amino acids) corresponding to the continuous components in the molecule from which the fragment is derived, and occupy at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% of the entire reference molecule from which the fragment is derived.

[0026] "Identity (sequence identity)": As used herein in the context of nucleic acid sequences or amino acid sequences, the term "identity" refers to the percentage of match between two sequences. To determine the percentage of identity between two sequences, the sequences are aligned (introducing gaps if necessary) and then compared to each other. In the context of the present invention, when a nucleotide is substituted by a modified nucleotide (e.g., when U is substituted by N1-methylpseudouridine (m1ψ)), it is not considered in the calculation of the percentage of identity. The percentage of identity between two sequences can be measured using, for example, an algorithm (such as the algorithm incorporated in the BLAST program).

[0027] "Immunogen", "immunogenicity": As used herein, the term "immunogen" or "immunogenicity" refers to a compound that can stimulate / induce an immune response. Preferably, an immunogen is a peptide, polypeptide, or protein.

[0028] "RNA in vitro transcription": The term "RNA in vitro transcription" or "in vitro transcription" refers to the process by which RNA is synthesized in an in vitro cell-free system. In RNA in vitro transcription (IVT), RNA is obtained by DNA-dependent in vitro transcription using a suitable DNA template in the presence of DNA-dependent RNA polymerase (such as T7, T3, SP6, or Syn5), ribonucleotide triphosphates (NTPs, and modified NTPs as needed); optionally, cap analogs, MgCl2, and buffer (containing TRIS or HEPES, and optionally DTT and / or spermidine).

[0029] Variant (Sequence Variant): In this specification, the term “variant” as used in the context of nucleic acid sequences is intended to be as recognized and understood by those skilled in the art, and is intended to refer, for example, to a variant of a nucleic acid sequence derived from another nucleic acid sequence. For example, a variant of a nucleic acid sequence may show one or more nucleotide deletions, insertions, additions, and / or substitutions compared to the nucleic acid sequence from which the variant is derived. A variant of a nucleic acid sequence may be at least 50%, 60%, 70%, 80%, 90%, or 95% identical to the nucleic acid sequence from which the variant is derived. A variant may be a functional variant in the sense that it retains at least 50%, 60%, 70%, 80%, 90%, or 95%, or more, of the function of the sequence from which the variant 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 region of at least 30, 50, 75, or 100 nucleotides of the nucleic acid sequence.

[0030] In this specification, the term “variant” as used in the context of proteins or peptides is intended to mean a protein or peptide variant having an amino acid sequence different from the original sequence due to one or more mutations / substitutions (one or more substitutions, insertions, and / or deletions of amino acids). Preferably, these fragments and / or variants have the same or equivalent specific properties. In particular, insertions and substitutions are possible at sequence locations that do not cause changes in the three-dimensional structure or affect binding regions. Changes in the three-dimensional structure due to insertions or deletions can be easily determined, for example, using a CD spectrum (circular dichroism spectrum). A protein variant may also be a functional variant of the protein from which it originates, meaning that the variant is essentially identical in function to the protein from which it originates, or exhibits at least 40%, 50%, 60%, 70%, 80%, or 90% of that function. A "variant" of a protein or peptide may have at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% amino acid identity in a region of at least 30, 50, 75, or 100 amino acids of the protein or peptide.

[0031] "NSCLC / Non-Small Cell Lung Cancer / Lung Cancer": Lung cancer, also known as lung carcinoma, is a malignant tumor that originates from lung cells. At diagnosis, lung cancer is classified based on the type of cells from which the tumor originates; tumor progression and response to treatment differ depending on the type of cells the tumor originates from. Approximately 85% of lung cancers are non-small cell lung cancers, including adenocarcinoma, squamous cell carcinoma, and large cell carcinoma. NSCLC can be classified into different subgroups (such as lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) / squamous non-small cell lung cancer (sqNSCLC)).

[0032] "Head and neck cancer": This is cancer that typically originates from the tissues of the lips and mouth, larynx (throat), salivary glands, nose, sinuses, or facial skin. The most common types of head and neck cancer occur in the lips, mouth, and larynx. Approximately 90% are pathologically classified as squamous cell carcinoma (HNSCC).

[0033] "Squamous cell carcinoma": Squamous cell carcinoma is a type of cancer that typically involves several different cancers that originate in squamous epithelial cells. These cells form on the surface of the skin, on the inner walls of cavity organs in the body, and on the inner walls of the respiratory and digestive tracts. Examples of squamous cell carcinomas include head and neck squamous cell carcinoma (cancers of the mouth, nasal cavity, nasopharynx, throat, and related structures), cutaneous squamous cell carcinoma, primary thyroid squamous cell carcinoma, esophageal squamous cell carcinoma, lung squamous cell carcinoma, and squamous cell carcinomas of the penis, vagina, ovaries, bladder, and eyes.

[0034] Melanoma is a type of skin cancer that arises from pigment-producing cells called melanocytes, and is one of the most dangerous types of skin cancer. The prognosis for melanoma depends on the stage at which it is discovered. Types of melanoma include, for example, cutaneous melanoma (SKCM) or uveal melanoma. Typically, SKCM can be classified into different subgroups (such as the BRAF subgroup and the NRAS subgroup).

[0035] [Detailed description of the invention] Where a “Sequence Number” is referred to in another patent application or patent, the sequence (such as an amino acid sequence or nucleic acid sequence) is expressly incorporated by reference herein. For “Sequence Numbers” described herein, the information listed in the “feature key” column in the sequence list according to the WIPO ST.26 standard, namely “source” (for nucleic acids or proteins), “misc_feature” (for nucleic acids), or “REGION” (for proteins), is also expressly included herein. Where a “Sequence Number” is referred to in the context of an RNA sequence, a person skilled in the art can obtain the RNA sequence from the referenced Sequence Number, even if a DNA sequence is provided. Where a “Sequence Number” is referred to in the context of a DNA sequence, a person skilled in the art can obtain the DNA sequence from the referenced Sequence Number, even if an RNA sequence is provided.

[0036] 1: Nucleic acid encoding at least one tumor antigen: In a first embodiment, the present invention provides a nucleic acid encoding at least one tumor antigen.

[0037] In particular, the specific features and embodiments described in the first aspect of the present invention, i.e., in the context of nucleic acids of the present invention, may also be applied to any aspect of the present invention, including the pharmaceutical composition described in the second aspect, the tumor antigen or composition of tumor antigen described in the third aspect, the antibody, T cell, or TCR described in the fourth aspect, the combination described in the fifth aspect, the kit or component kit described in the sixth aspect, or any aspect relating to medical use or therapeutic methods.

[0038] In a preferred embodiment, the nucleic acid encoding at least one tumor antigen is an artificial nucleic acid.

[0039] As used herein, the term “artificial nucleic acid” refers to nucleic acids that do not exist in nature. In other words, artificial nucleic acids may be understood as non-natural nucleic acid molecules. Such nucleic acid molecules may be non-natural by their individual sequences (e.g., codon-modified coding sequences, UTRs) and / or other modifications (e.g., structural changes of nucleotides). Typically, artificial nucleic acids may be designed and / or fabricated by genetic engineering to correspond to a desired artificial nucleotide sequence. In this context, an artificial nucleic acid is a sequence that does not exist in nature, i.e., a sequence that differs from a wild-type sequence or a sequence found in nature by at least one nucleotide. The term “artificial nucleic acid” is not limited to “a single molecule” and is understood to include a collection of essentially identical nucleic acid molecules. As used herein, the term “artificial nucleic acid” may refer to artificial DNA or artificial RNA. Preferably, the artificial nucleic acid is selected from artificial RNA.

[0040] In a preferred embodiment, the artificial nucleic acid includes at least one coding sequence encoding at least one tumor antigen.

[0041] As used herein, the term "tumor antigen" is intended to refer to tumor-associated antigens (TAAs) and tumor-specific antigens (TSAs) (including covalent antigens, neoantigens, and specific antigens) that are expressed in both normal and tumor tissues. Furthermore, tumor antigens can be broadly classified into the following categories: oncofetal (usually expressed only in fetal tissue and cancerous somatic cells), oncoviral (encoded by tumorigenic transforming virus), overexpression / accumulative (expressed in both normal and tumor tissue, but with significantly increased expression levels in tumors), cancer-testicular (expressed only in adult reproductive tissues such as the testes and placenta, and in cancer cells), lineage-restricted (expressed mainly in a single type of cancer tissue), mutant (expressed only in cancer due to gene mutations or transcriptional changes), post-translational modification (such as tumor-associated changes in glycosylation), or idiotype (highly polymorphic genes in which tumor cells express a specific "clonal type," i.e., B-cell or T-cell lymphoma / leukemia caused by clonal abnormalities). It should be emphasized that these classifications are not mutually exclusive, and tumor antigens may fall into multiple categories.

[0042] In a preferred embodiment, tumor antigens (obtained by artificial nucleic acids) are produced in the cytosol upon administration of the artificial nucleic acids to cells, tissues, or a target. The produced tumor antigens may be secreted or further processed.

[0043] Therefore, when an artificial nucleic acid (e.g., RNA) is administered to cells, tissues, or subjects, at least one coding sequence is translated into at least one tumor antigen. When referring to “produced tumor antigen,” this term refers to the protein product generated from the artificial nucleic acid of the present invention by translating the coding sequence of the nucleic acid into a protein. Accordingly, functional and structural features, as well as embodiments, described herein in relation to “tumor antigen” or “(produced) tumor antigen” should be understood to refer to the tumor antigen amino acid sequence produced / translated in the cytosol upon administration of the artificial nucleic acid to cells, tissues, or subjects.

[0044] In one embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is At least one antigenic peptide selected from, or derived from, a peptide or protein encoded by a long non-coding RNA (lncRNA), The immunogenic fragment or variant of the peptide or protein, It includes or consists of.

[0045] In one embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is At least one antigenic peptide selected from or derived from a tumor neoantigen peptide or protein, The immunogenic fragment or variant of the peptide or protein, It includes or consists of.

[0046] The terms “immunogenic fragment” or “immunogenic variant” should be understood as any fragment / variant of a corresponding peptide or protein that has the ability to induce an immune response in a subject, preferably a human subject.

[0047] In one embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is a) at least one amino acid sequence encoded by a long non-coding RNA (lncRNA), or an immunogenic fragment thereof or a variant thereof; and / or b) At least one amino acid sequence of a tumor neoantigen, or an immunogenic fragment or variant thereof, It includes or consists of.

[0048] In one embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is a) A peptide or protein encoded by a long non-coding RNA (lncRNA), or at least one antigenic peptide selected from or derived from an immunogenic fragment or variant thereof; and / or b) A peptide or protein of a tumor neoantigen, or at least one antigenic peptide selected from or derived from an immunogenic fragment or variant thereof, It includes or consists of.

[0049] In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is A peptide or protein encoded by a long non-coding RNA (lncRNA), or at least one antigenic peptide selected from or derived from an immunogenic fragment or variant thereof, It includes or consists of.

[0050] [Tumor antigens encoded by long non-coding RNAs (lncRNAs)] In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is A peptide or protein encoded by a long non-coding RNA (lncRNA), or at least one antigenic peptide selected from or derived from an immunogenic fragment or variant thereof, It includes or consists of.

[0051] The term "long non-coding RNA" and its abbreviation "lncRNA" typically refer to RNA with a length of 200 nucleotides or more (e.g., up to 10.0000 nucleotides). Traditionally, lncRNAs were thought to have no protein-coding function. However, recent research on long non-coding RNAs has revealed that these molecules are involved in diverse regulatory mechanisms of cells. Compared to mRNA, long non-coding RNAs are characterized by lower expression levels, but exhibit strong specificity in tissue and cell expression. Recently, it has been discovered that several lncRNAs encode peptides with biologically important functions (micropeptides or putative small open reading frames (also referred to herein as "mini ORF" or "small ORF" (smORF))). Therefore, based on the nomenclature common in the art, the term "long non-coding RNA" or its abbreviation "lncRNA" will be used consistently within this specification. In particular, the lncRNA species in the context of the present invention encode peptides or proteins that can function as appropriate tumor antigens.

[0052] Remarkably, the inventors identified peptides produced by specific lncRNA species. Even more surprisingly, these peptides encoded by lncRNAs exhibit specificity for certain tumors (melanoma, NSCLC, HNSCC, etc.) and can trigger antigen-specific immune responses. For example, immune cells may recognize and present these peptides as foreign substances, triggering an antigen-specific immune response (e.g., a T-cell response). Therefore, peptides selected from or derived from lncRNA smORFs are suitable for cancer immunotherapy.

[0053] In a preferred embodiment, at least one antigenic peptide is The amino acid sequence of a peptide or protein encoded by long non-coding RNA (lncRNA), or The (immunogenic) fragment or its variant, Selected from the following, where the lncRNA comprises at least one smORF (encoding the peptide or protein), wherein the lncRNA is characterized by increased expression in cancer cells (melanoma, NSCLC, HNSCC, etc.) compared to non-cancer cells. Furthermore, the lncRNA comprises at least one smORF.

[0054] In a preferred embodiment, at least one antigenic peptide is selected from a peptide or protein encoded by a long non-coding RNA (lncRNA), or from an immunogenic fragment thereof or a variant thereof. 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 from any immunogenic fragment or variant thereof.

[0055] In a preferred embodiment, at least one antigenic peptide comprises an amino acid sequence encoded by a long non-coding RNA (lncRNA), or a fragment or variant thereof. 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 selected from any immunogenic fragment or variant thereof.

[0056] In the context of specific lncRNA species (e.g., those encoded by ZC3H8-6:1, WDR72-2:4, KCNMB2-AS1:4, or NTF3-5:5), the terms "amino acid sequence encoded by ~," "peptide or protein encoded by ~," or "antigen encoded by ~" define the origin or source of each amino acid sequence, peptide, protein, or antigen as a long noncoding RNA. In other words, each amino acid sequence, peptide, protein, or antigen originates from its respective lncRNA species. In the context of the present invention, each amino acid sequence, peptide, protein, or antigen encoded by a specific lncRNA species (such as ZC3H8-6:1, WDR72-2:4, KCNMB2-AS1:4, or NTF3-5:5), or in other words, derived from a specific lncRNA species (such as ZC3H8-6:1, WDR72-2:4, KCNMB2-AS1:4, or NTF3-5:5), is provided by the nucleic acid (e.g., mRNA) of the present invention.

[0057] In the context of the present invention, preferred is, Amino acid sequences of peptides or proteins encoded by lnc-NTF3-5:5, lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-ZC3H8-6:1, or lnc-CLEC2D-9:1, or immunogenic fragments or variants thereof, or A nucleic acid sequence encoding an amino acid sequence such as a peptide or protein encoded by lnc-NTF3-5:5, lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-ZC3H8-6:1, or lnc-CLEC2D-9:1, or a fragment or variant thereof.

[0058] In the context of the present invention, particularly preferred is, The amino acid sequence of a peptide or protein encoded by lnc-NTF3-5:5, or its immunogenic fragment or variant, or A nucleic acid sequence that encodes an amino acid sequence such as a peptide or protein encoded by lnc-NTF3-5:5, or a fragment or variant thereof.

[0059] Importantly, 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, as indicated by RNA sequencing (see Examples section). Furthermore, the peptides encoded by these lncRNAs have been identified by mass spectrometry (see Examples section).

[0060] lnc-WDR72-2:4 is a long non-coding RNA transcribed in humans from the following genomic locations: chr15,53513740,53540852,-. The naturally occurring (human) lncRNA sequence of lnc-WDR72-2:4 corresponds to Sequence ID No. 512. The RNA sequence of lnc-WDR72-2:4 comprises at least two open reading frames (smORF lncWDR722416421696 and lncWDR722415641696) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by lnc-WDR72-2:4, or its immunogenic fragment or variant, can function as a suitable tumor antigen in the context of the present invention.

[0061] 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 corresponds to SEQ ID NO: 516. The RNA sequence of lnc-TRPC5-3:1 includes at least one open reading frame (smORF lncTRPC53116031801) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by lnc-TRPC5-3:1, or its immunogenic fragment or variant, can function as a suitable tumor antigen in the context of the present invention.

[0062] 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 corresponds to Sequence ID No. 517. The RNA sequence of lnc-CLEC2D-9:1 includes at least one open reading frame (smORF lncCLEC2D91412616) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by lnc-CLEC2D-9:1, or its immunogenic fragment or variant, can function as a suitable tumor antigen in the context of the present invention.

[0063] KCNMB2-AS1:4 is a long non-coding RNA transcribed in humans from the following genomic location: chr3,178525486,178860397,-. The naturally occurring (human) lncRNA sequence of KCNMB2-AS1:4 corresponds to Sequence ID No. 513. The RNA sequence of KCNMB2-AS1:4 includes at least one open reading frame (smORF KCNMB2AS14633753) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by KCNMB2-AS1:4, or its immunogenic fragment or variant, can function as a suitable tumor antigen in the context of the present invention.

[0064] lnc-ZC3H8-6:1 is a long non-coding RNA transcribed in humans from the following genomic location: chr2,111769354,111776910,-. The naturally occurring (human) lncRNA sequence of lnc-ZC3H8-6:1 corresponds to Sequence ID No. 514. The RNA sequence of lnc-ZC3H8-6:1 contains at least two open reading frames (smORF lncZC3H86163144) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by lnc-ZC3H8-6:1, or its immunogenic fragment or variant, can function as a suitable tumor antigen in the context of the present invention.

[0065] lnc-NTF3-5:5 is a long non-coding RNA transcribed in humans from the following genomic locations: chr12,5367810,5379568,+. The naturally occurring (human) lncRNA sequence of lnc-NTF3-5:5 corresponds to SEQ ID NO: 515. The RNA sequence of lnc-NTF3-5:5 contains at least two open reading frames (smORF lncNTF35511951417) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by lnc-NTF3-5:5, or its immunogenic fragment or variant thereof, can function as a suitable tumor antigen in the context of the present invention.

[0066] LINC00893:25 (encoding smORF LINC00893) is a long non-coding RNA transcribed in humans from the following genomic location: chrX,149483112,149532914,-. The naturally occurring (human) lncRNA sequence of LINC00893:25 corresponds to Sequence ID No. 518. The RNA sequence of LINC00893:25 comprises at least two open reading frames (smORF LINC00893) encoding at least one amino acid sequence (e.g., at least one peptide that can function as a tumor antigen in the context of the present invention). Preferably, any peptide or protein produced by LINC00893:25, or its immunogenic fragment or variant, can function as a suitable tumor antigen in the context of the present invention.

[0067] Preferably, at least one antigenic peptide encoded by a long non-coding RNA (lncRNA), or any immunogenic fragment or variant thereof, comprises at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0068] Preferably, at least one antigenic peptide encoded by the 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 immunogenic fragment or variant thereof, comprises at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0069] In a preferred embodiment, at least one antigenic peptide encoded by a long non-coding RNA (lncRNA), or any immunogenic fragment or variant thereof, induces epitope-specific T cells, preferably epitope-specific CD8+ T cells (particularly in human subjects), in a subject.

[0070] In a particularly preferred embodiment, at least one antigenic peptide encoded by the 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 immunogenic fragment or variant thereof, induces epitope-specific T cells, preferably epitope-specific CD8+ T cells (particularly in human subjects) in a subject.

[0071] 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 (MHC) molecules. It also refers to gene complexes present in all vertebrates, including MHC class I and MHC class II molecules. MHC proteins or molecules are crucial for signal transduction between lymphocytes and antigen-presenting or diseased cells in immune responses, where they bind to peptide epitopes and present them for recognition by T cell receptors on T cells. The MHC-encoded proteins are expressed on the cell surface and present both self-antigens (peptide fragments of the cell itself) and non-self-antigens (fragments of invading microorganisms, etc.) to T cells. In the case of class I MHC / peptide complexes, the binding peptide (MHC class I ligand) is typically about 8 to 10 amino acids long, although longer or shorter peptides may also be effective. In the case of class II MHC / peptide complexes, the binding peptide (MHC class II ligand) is typically about 10 to 25 amino acids long, especially about 13 to 18 amino acids long, but longer or shorter peptides may also be effective.

[0072] Preferably, at least one antigenic peptide encoded by an 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.

[0073] As used herein, the terms "HLA" or "human leukocyte antigen" include HLA variants, isoforms, and interspecies homologs, as well as analogs that share at least one common epitope with HLA. As used herein, the term "human leukocyte antigen (HLA)" should preferably be understood as the gene complex encoding human "major histocompatibility complex (MHC)" proteins. These cell surface proteins are responsible for regulating the immune system in humans. HLA genes are highly polymorphic, meaning they have many different alleles, which allows for precise tuning of the adaptive immune system of a given subject. As used herein, the terms "HLA binding affinity" or "MHC binding affinity" are understood as the binding affinity between a particular antigen and a particular MHC allele. As used herein, the term "HLA type" is understood as a combination (complement) of HLA gene alleles.

[0074] Preferably, at least one antigenic peptide encoded by an lncRNA as defined herein can bind to an antibody or T cell receptor. Preferably, the antigenic peptide comprises at least one MHC I ligand or MHC II ligand.

[0075] In embodiments, at least one antigenic peptide encoded by an lncRNA as defined herein is a covalent tumor-associated antigen, particularly a covalent tumor-associated antigen in NSCLC and / or melanoma.

[0076] In a preferred embodiment, at least one antigenic peptide (obtained by an artificial nucleic acid) is selected from peptides or proteins encoded by long non-coding RNA (lncRNA) as defined herein. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 124-254, 546-650, or 756-763, or at least one that is identical to any one of these by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those immunogenic fragments or any of those variants, It includes or consists of.

[0077] Therefore, at least one antigenic peptide is preferably, At least one amino acid sequence that is identical to any one of sequence numbers 124-254, 546-650, or 756-763, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, It includes or consists of.

[0078] In this context, the immunogenic fragments of SEQ ID NOs. 124-254, 546-650, and 756-763 have shorter amino acid sequences than their respective reference sequences. Therefore, the immunogenic fragments of SEQ ID NOs. 124-254, 546-650, and 756-763 are shortened by (e.g., 1, 5, 10 or more) amino acids at the N-terminus and / or by (e.g., 1, 5, 10 or more) amino acids at the C-terminus. In particular, the immunogenic fragments still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In embodiments, at least one epitope has the size / length of an MHC class I or class II epitope. In preferred embodiments, 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 includes at least one MHC class I epitope, and can be processed during antigen treatment in the loading process of the MHC:peptide complex. In this context, typical immunogenic fragments have a size of 5 to 20 amino acids or 8 to 15 amino acids.

[0079] (lnc-WDR72-2:4) 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 thereof or a variant thereof.

[0080] The full-length polypeptide encoded by lncRNA lnc-WDR72-2:4 (SEQ ID NOs. 124 and 144) contains multiple epitopes predicted based on SEQ ID NOs. 125-143, 145-146, and 546-571. Therefore, the full-length polypeptide (smORF lncWDR722416421696 or lncWDR722415641696 based on SEQ ID NOs. 124 and 144), or any fragment or variant thereof containing the epitopes defined herein, is particularly useful in the context of the present invention.

[0081] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-WDR72-2:4. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 124-146, 546-571, 756, or 757, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0082] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 124-146, 546-571, 756, or 757, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0083] In this context, the fragments may be N-terminal truncations of SEQ ID NOs. 124-146, 546-571, 756, and 757, or C-terminal truncations of SEQ ID NOs. 124-146, 546-571, 756, and 757. In embodiments, the immunogenic fragments of SEQ ID NOs. 124-146, 546-571, 756, and 757 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. 124-146, 546-571, 756, and 757 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0084] In a particularly preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-WDR72-2:4. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 756, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to it, The immunogenic fragment or its immunogenic variant, It includes or consists of.

[0085] (lnc-TRPC5-3:1) In embodiments, at least one antigenic peptide comprises an amino acid sequence (such as a peptide or protein encoded by lnc-TRPC5-3:1 (smORF lncTRPC53116031801)) or an immunogenic fragment thereof or a variant thereof.

[0086] The full-length polypeptide encoded by lncRNA lnc-TRPC5-3:1 (SEQ ID NO: 227) contains multiple epitopes predicted based on SEQ ID NOs: 228-237, 630-642. Therefore, the full-length polypeptide (smORF lncTRPC53116031801 based on SEQ ID NO: 227), or any fragment thereof or variants thereof containing the epitopes defined herein, are particularly useful in the context of the present invention.

[0087] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-TRPC5-3:1. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 227-237, 630-642, or 761, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0088] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 227-237, 630-642, or 761, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0089] In this context, the fragments may be N-terminal abbreviations of SEQ ID NOs. 227-237, 630-642, and 761, or C-terminal abbreviations of SEQ ID NOs. 227-237, 630-642, and 761. In embodiments, the immunogenic fragments of SEQ ID NOs. 227-237, 630-642, and 761 are abbreviated 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. 227-237, 630-642, and 761 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0090] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by lnc-TRPC5-3:1. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 761, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or The immunogenic fragment or its immunogenic variant, It includes or consists of.

[0091] (lnc-CLEC2D-9:1) 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 thereof or an immunogenic variant thereof.

[0092] The full-length polypeptide encoded by lncRNA lnc-CLEC2D-9:1 (SEQ ID NO: 238) contains multiple epitopes predicted based on SEQ ID NOs: 239-243, 643-650. Therefore, the full-length polypeptide (smORF lncCLEC2D91412616 based on SEQ ID NO: 238), or any fragment thereof or variant thereof containing the epitopes defined herein, is particularly useful in the context of the present invention.

[0093] In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by lnc-CLEC2D-9:1. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 238-243, 643-650, or 762, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0094] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 238-243, 643-650, or 762, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0095] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 238-243, 643-650, and 762, or C-terminal shortened versions of SEQ ID NOs. 238-243, 643-650, and 762. In embodiments, the immunogenic fragments of SEQ ID NOs. 238-243, 643-650, and 762 are shortened 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. 238-243, 643-650, and 762 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0096] In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by lnc-CLEC2D-9:1. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 762, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to it, The immunogenic fragment or its immunogenic variant, It includes or consists of.

[0097] (KCNMB2-AS1:4) 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 thereof or an immunogenic variant thereof.

[0098] The full-length polypeptide encoded by lncRNA KCNMB2-AS1:4 (SEQ ID NO: 147) contains multiple epitopes predicted based on SEQ ID NOs: 148-161, 572-589. Therefore, the full-length polypeptide (smORF KCNMB2AS14633753 based on SEQ ID NO: 147), or any fragment thereof or variants thereof containing the epitopes defined herein, are particularly useful in the context of the present invention.

[0099] In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by KCNMB2-AS1:4. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 147-161, 572-589, or 758, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0100] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 147-161, 572-589, or 758, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0101] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs: 147-161, 572-589, and 758, or C-terminal shortened versions of SEQ ID NOs: 147-161, 572-589, and 758. In embodiments, the immunogenic fragments of SEQ ID NOs: 147-161, 572-589, and 758 are shortened 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: 147-161, 572-589, and 758 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0102] In a particularly preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by KCNMB2-AS1:4. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 758, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0103] (lnc-ZC3H8-6:1) 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 thereof or a variant thereof.

[0104] The full-length polypeptide encoded by lncRNA lnc-ZC3H8-6:1 (SEQ ID NO: 162) contains multiple epitopes predicted based on SEQ ID NOs: 163-168, 590-603. Therefore, the full-length polypeptide (smORF lncZC3H86163144 based on SEQ ID NO: 162), or any fragment thereof or variant thereof containing the epitopes defined herein, is particularly useful in the context of the present invention.

[0105] In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by lnc-ZC3H8-6:1. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 162-168, 590-603, or 759, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0106] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 162-168, 590-603, or 759, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0107] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 162-168, 590-603, and 759, or C-terminal shortened versions of SEQ ID NOs. 162-168, 590-603, and 759. In embodiments, the immunogenic fragments of SEQ ID NOs. 162-168, 590-603, and 759 are shortened 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. 162-168, 590-603, and 759 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0108] In a particularly preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by lnc-ZC3H8-6:1. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 759, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to it, The immunogenic fragment or its immunogenic variant, It includes or consists of.

[0109] (lnc-NTF3-5:5) 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 thereof or a variant thereof.

[0110] The full-length polypeptide encoded by lncRNA lnc-NTF3-5:5 (SEQ ID NO: 169) contains multiple epitopes predicted based on SEQ ID NOs: 170-226, 604-629. Therefore, the full-length polypeptide (smORF lncNTF35511951417 based on SEQ ID NO: 169), or any fragment thereof or variant thereof containing the epitopes defined herein, is particularly useful in the context of the present invention.

[0111] In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by lnc-NTF3-5:5. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 169-226, 604-629, or 760, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0112] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 169-226, 604-629, or 760, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0113] In this context, the fragments may be N-terminal abbreviations of SEQ ID NOs. 169-226, 604-629, and 760, or C-terminal abbreviations of SEQ ID NOs. 169-226, 604-629, and 760. In embodiments, the immunogenic fragments of SEQ ID NOs. 169-226, 604-629, and 760 are shortened 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. 169-226, 604-629, and 760 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0114] In a particularly preferred embodiment, at least one antigenic peptide is selected from the peptide or protein encoded by lnc-NTF3-5:5. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 760, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to it, The immunogenic fragment or its immunogenic variant, It includes or consists of.

[0115] (LINC00893:25) In embodiments, 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 thereof or a variant thereof.

[0116] The full-length polypeptide (smORF LINC00893 based on SEQ ID NO: 244), or any fragment thereof containing the epitopes defined herein, or any variant thereof, is particularly useful in the context of the present invention. Accordingly, the full-length polypeptide encoded by lncRNA LINC00893 (SEQ ID NO: 244) contains multiple epitopes predicted based on SEQ ID NOs: 245-254.

[0117] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by LINC00893:25. The antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 244-254 or 763, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0118] Therefore, preferably at least one antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 244-254 or 763, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0119] In this context, the fragments may be N-terminal abbreviations of SEQ ID NOs. 244-254, 763, or C-terminal abbreviations of SEQ ID NOs. 244-254, 763. In embodiments, the immunogenic fragments of SEQ ID NOs. 244-254, 763 are shortened 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. 244-254, 763 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope. In this context, typical immunogenic fragments have a size of 5-20 amino acids or 8-15 amino acids.

[0120] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by LINC00893:25. The antigenic peptide is At least one amino acid sequence that is identical to sequence number 763, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to it, or The immunogenic fragment or its immunogenic variant, It includes or consists of.

[0121] (Preferred nucleic acid sequence encoding an antigen encoded by lncRNA) The following describes nucleic acid coding sequences that encode specific antigenic peptides (encoded by their respective lncRNAs) as used herein. These sequences may be contained within at least one cds of an artificial nucleic acid.

[0122] In a preferred embodiment, at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from a peptide or protein encoded by an lncRNA, wherein the nucleic acid sequence is It is identical to any one of sequence numbers 318-448, 651-755, or 764-771, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments or any of those variants, Preferably, in this context, the fragment has a length of at least 24 nucleotides.

[0123] Preferably, the nucleic acid sequence encoding at least one antigenic peptide selected from the peptides or proteins encoded by the lncRNA is a codon-modified cds. Preferably, the codon-modified cds are selected from C-maximized cds (described below), CAI-maximized cds (described below), cds adapted for human codon use (described below), G / C content modified cds (described below), and G / C optimized cds (described below), or any combination thereof. In the preferred embodiment in this context, at least one codon-modified coding sequence is a G / C optimized coding sequence.

[0124] 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 a peptide or protein encoded by an lncRNA, wherein the nucleic acid sequence is It is identical to any one of sequence numbers 318-448, 651-755, or 764-771, or it is at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or It is one of those fragments. In this context, preferably, the fragment has a length of at least 24 nucleotides.

[0125] In a particularly preferred embodiment, at least one artificial nucleic acid includes: i) at least one cds comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from the peptide or protein encoded by lnc-NTF3-5:5, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 363-420, 709-734, or 768, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds, which are sequence number 768 or a fragment thereof; or, ii) At least one CDS comprising a nucleic acid sequence encoding at least one tumor antigen, which includes at least one antigenic peptide selected from peptides or proteins encoded by lnc-WDR72-2:4, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 318-340, 651-676, 764, or 765, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds are sequence number 765 or a fragment thereof; or, iii) at least one cds comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-TRPC5-3:1, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 421-431, 735-747, or 769, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds are sequence number 769 or a fragment thereof; or, iv) at least one cds comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from the peptide or protein encoded by lnc-CLEC2D-9:1, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 432-437, 748-755, or 770, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds, which are sequence number 770 or a fragment thereof; or v) At least one cds comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from the peptide or protein encoded by KCNMB2-AS1:4, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 341-355, 677-694, or 766, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds, which are sequence number 766 or a fragment thereof; or, vi) At least one cds comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from a peptide or protein encoded by lnc-ZC3H8-6:1, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 356-362, 695-708, or 767, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds, which are sequence number 767 or a fragment thereof; or, vii) At least one cds comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from peptides or proteins encoded by LINC00893:25, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 438-448 or 771, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these, or It is a fragment of any of those or a variant of any of those. Preferably, cds are sequence number 771 or a fragment thereof or a variant thereof.

[0126] Table 1A shows preferred amino acid and nucleic acid sequences in this context. Each row corresponds to a suitable antigenic peptide or antigenic protein encoded by a long non-coding RNA (lncRNA). Column A provides a brief description of each lncRNA encoding a tumor antigen. Column B shows the sequence number of each full-length amino acid sequence encoded by the lncRNA. Column C shows the amino acid sequence number of each fragment or epitope. Column D shows the sequence number of the G / C optimized nucleic acid sequence encoding the full-length amino acid sequence in column B. Column E shows the sequence number of the G / C optimized nucleic acid sequence encoding each fragment or epitope in column C. Column F indicates the main cancer species to which the antigen is applicable (N=non-small cell lung cancer; M=melanoma; H=head and neck squamous cell carcinoma; see also Embodiment 7 (Medical Uses) for more detailed cancer species to which the antigen is applicable, in particular for antigens encoded by lncRNA (listed in rows 1-4 and 6)).

[0127] [Table 1]

[0128] Table 1B shows particularly preferred amino acid and nucleic acid sequences in this context. Each row corresponds to a particularly suitable antigenic peptide or antigenic protein encoded by a long non-coding RNA (lncRNA). Column A provides a brief description of each lncRNA encoding a tumor antigen. Column B shows the sequence number of each full-length amino acid sequence encoded by the lncRNA. Column C shows the amino acid sequence number of a preferred fragment or epitope. Column D shows the sequence number of a G / C optimized nucleic acid sequence encoding the full-length amino acid sequence in column B. Column E shows the sequence number of a G / C optimized nucleic acid sequence encoding a preferred fragment or epitope in column C.

[0129] [Table 2]

[0130] In a particularly preferred embodiment, the antigenic peptide comprises or consists of at least one amino acid sequence selected from column B or C of Table 1A or Table 1B, or an immunogenic fragment thereof.

[0131] In a particularly preferred embodiment, at least one coding sequence of the artificial nucleic acid of the present invention comprises or consists of at least one amino acid sequence selected from column D or E of Table 1A or Table 1B, or an immunogenic fragment or immunogenic variant thereof.

[0132] [Tumor antigens selected from tumor neoantigens] In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence encoding at least one tumor antigen. The tumor antigen is A tumor neoantigen peptide or protein, or at least one antigenic peptide selected from or derived from its immunogenic fragment or variant, It includes or consists of.

[0133] The term "tumor neoantigen" refers to specific polypeptides produced by genetic mutations in tumor cells. These polypeptides are presented on the cell surface by MHC molecules (HLA molecules in mammals) and recognized by T lymphocytes, triggering an immune response. Therefore, neoantigens are suitable as markers for immunotherapy and are also used in the development of tumor neoantigen vaccines. This is because these neoantigens are produced by mutations in tumor cells but are not expressed in normal or healthy cells. Consequently, neoantigens have the potential to be promising targets for immunotherapy.

[0134] In a preferred embodiment, the neoantigen is a covalent neoantigen, preferably a covalent neoantigen common to patients with NSCLC and / or melanoma cancer.

[0135] Surprisingly, peptides present in tumor cells were identified. Even more surprisingly, these peptides exhibit specificity in tumor cells and trigger an immune response, preferably an antigen-specific immune response.

[0136] In this embodiment, at least one antigenic peptide selected from the tumor neoantigen peptide or protein 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 thereof.

[0137] For clarification, 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" are related to their respective mutant proteins present in cancer cells, and not to their respective wild-type or somatic proteins.

[0138] In a preferred embodiment, at least one antigenic peptide is selected from the 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 immunogenic fragments or variants thereof.

[0139] In a preferred embodiment, at least one antigenic peptide selected from the 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, and PATZ1 neoantigen comprises at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0140] In a particularly preferred embodiment, at least one antigenic peptide selected from tumor neoantigen peptides or proteins, or immunogenic fragments or variants thereof, particularly the antigenic peptides 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, and PATZ1 neoantigen, induces epitope-specific T cells, preferably epitope-specific CD8+ T cells (particularly in human subjects) in the subject.

[0141] In a preferred embodiment, at least one antigenic peptide selected from a tumor neoantigen peptide or protein, or an immunogenic fragment thereof or a variant thereof, is an antigen or epitope exhibiting HLA-binding affinity or MHC-binding affinity.

[0142] Preferably, at least one antigenic peptide of the tumor neoantigen as defined herein binds to an antibody or T cell receptor. In a preferred embodiment, the antigenic peptide comprises an MHC I or MHC II ligand / epitope.

[0143] In the embodiment, at least one antigenic peptide (obtained by artificial nucleic acid) is selected from a tumor neoantigen peptide or protein. At least one amino acid sequence that is identical to any one of sequence numbers 255-317, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, It includes or consists of.

[0144] In a preferred embodiment, at least one antigenic peptide (obtained by an artificial nucleic acid) is selected from a tumor neoantigen peptide or protein. At least one amino acid sequence that is identical to any one of sequence numbers 255-286, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, It includes or consists of.

[0145] In this context, the immunogenic fragments of SEQ ID NOs. 255–317 have shorter amino acid sequences than their respective reference sequences. The immunogenic fragments of SEQ ID NOs. 255–317 are shortened by (e.g., 1, 5, 10 or more) amino acids at the N-terminus and / or by (e.g., 1, 5, 10 or more) amino acids at the C-terminus. In particular, the immunogenic fragments still contain at least one epitope capable of inducing 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 class II epitope. In preferred embodiments, 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 that can be processed during antigen processing in the loading process of the MHC:peptide complex. In this context, typical immunogenic fragments have a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0146] (NRAS amino acid and nucleic acid sequences) In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of NRAS tumor neoantigens. At least one amino acid sequence that is identical to any one of sequence numbers 267-272, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptides or proteins of NRAS tumor neoantigens. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 267-272, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0147] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 267-272, or C-terminal shortened versions of SEQ ID NOs. 267-272. In embodiments, the immunogenic fragments of SEQ ID NOs. 267-272 are shortened 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. 267-272 still contain at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, typical immunogenic fragments have a size of 5-20 amino acids, or 8-15 amino acids.

[0148] (TYW1B amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the TYW1B tumor neoantigen. At least one amino acid sequence that is identical to either SEQ ID NOs. 273, 274, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these, or Those immunogenic fragments or their immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptide or protein of the TYW1B tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 273 or 274, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0149] In this context, the fragment may be an N-terminal shortened form of SEQ ID NO: 273,274, or a C-terminal shortened form of SEQ ID NO: 273,274. In embodiments, the immunogenic fragment of SEQ ID NO: 273,274 is shortened 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 fragment of SEQ ID NO: 273,274 still contains at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, a typical immunogenic fragment has a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0150] (ECPAS amino acid and nucleic acid sequences) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the ECPAS tumor neoantigen. At least one amino acid sequence that is identical to any of sequence number 275, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of these sequences, The immunogenic fragment or its immunogenic variant, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptides or proteins of ECPAS tumor neoantigens. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 275, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0151] In this context, the fragment may be an N-terminal abbreviation of SEQ ID NO: 275, or a C-terminal abbreviation of SEQ ID NO: 275. In embodiments, the immunogenic fragment of SEQ ID NO: 275 is abbreviated 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 fragment of SEQ ID NO: 275 still contains at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, a typical immunogenic fragment has a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0152] (MAP2K1 amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the MAP2K1 tumor neoantigen. At least one amino acid sequence that is identical to either SEQ ID NOs. 276 or 277, or at least one amino acid sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these, Those immunogenic fragments or their immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptide or protein of the MAP2K1 tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 276 or 277, or at least 80% identical to it, Those immunogenic fragments or their variants, It includes or consists of.

[0153] In this context, the fragment may be an N-terminal abbreviation of SEQ ID NO: 276,277, or a C-terminal abbreviation of SEQ ID NO: 276,277. In embodiments, the immunogenic fragment of SEQ ID NO: 276,277 is abbreviated 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 fragment of SEQ ID NO: 276,277 still contains at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, a typical immunogenic fragment has a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0154] (TOMM22 amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the TOMM22 tumor neoantigen. At least one amino acid sequence that is identical to either SEQ ID NOs. 278, 279, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these, or Those immunogenic fragments or their immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptide or protein of the TOMM22 tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 278 or 279, or at least 80% identical to it, Those immunogenic fragments or their variants, It includes or consists of.

[0155] In this context, the fragment may be an N-terminal abbreviation of SEQ ID NO: 278,279, or a C-terminal abbreviation of SEQ ID NO: 278,279. In embodiments, the immunogenic fragment of SEQ ID NO: 278,279 is abbreviated 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 fragment of SEQ ID NO: 278,279 still contains at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, a typical immunogenic fragment has a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0156] (GLB1 amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of the GLB1 tumor neoantigen. At least one amino acid sequence that is identical to either SEQ ID NOs. 280 or 281, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptide or protein of the GLB1 tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NO: 280 or 281, or at least 80% identical to it, Those immunogenic fragments or their variants, It includes or consists of.

[0157] In this context, the fragment may be an N-terminal shortened form of SEQ ID NO: 280,281, or a C-terminal shortened form of SEQ ID NO: 280,281. In embodiments, the immunogenic fragment of SEQ ID NO: 280,281 is shortened 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 fragment of SEQ ID NO: 280,281 still contains at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, a typical immunogenic fragment has a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0158] (MAGE-A3 amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the MAGE-A3 tumor neoantigen. At least one amino acid sequence that is identical to either SEQ ID NOs. 282 or 283, or at least one amino acid sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these, or Those immunogenic fragments or their immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptides or proteins of the MAGE-A3 tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 282 or 283, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0159] In this context, the fragment may be an N-terminal shortened form of SEQ ID NO: 282,283, or a C-terminal shortened form of SEQ ID NO: 282,283. In embodiments, the immunogenic fragment of SEQ ID NO: 282,283 is shortened 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 fragment of SEQ ID NO: 282,283 still contains at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, a typical immunogenic fragment has a size of 5 to 20 amino acids, or 8 to 15 amino acids.

[0160] (ATAD2 amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the ATAD2 tumor neoantigen. At least one amino acid sequence that is identical to any one of sequence numbers 284-286, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Those immunogenic fragments or their immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptide or protein of the ATAD2 tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 284-286, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0161] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 284-286, or C-terminal shortened versions of SEQ ID NOs. 284-286. In embodiments, the immunogenic fragments of SEQ ID NOs. 284-286 are shortened 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. 284-286 still contain at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response. In this context, typical immunogenic fragments have a size of 5-20 amino acids, or 8-15 amino acids.

[0162] (BRAF amino acid and nucleic acid sequences) In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins of BRAF tumor neoantigens. At least one amino acid sequence that is identical to any one of sequence numbers 262-266, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptides or proteins of BRAF tumor neoantigens. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 262-266, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0163] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 262-266, or C-terminal shortened versions of SEQ ID NOs. 262-266. In embodiments, the immunogenic fragments of SEQ ID NOs. 262-266 are shortened 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. 262-266 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope.

[0164] (EGFR amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the EGFR tumor neoantigen. At least one amino acid sequence that is identical to any one of sequence numbers 255-257, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptides or proteins of EGFR tumor neoantigens. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 255-257, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0165] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 255-257, or C-terminal shortened versions of SEQ ID NOs. 255-257. In embodiments, the immunogenic fragments of SEQ ID NOs. 255-257 are shortened 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. 255-257 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope.

[0166] (TP53 amino acid and nucleic acid sequence) In a preferred embodiment, at least one antigenic peptide is selected from the peptide or protein of the TP53 tumor neoantigen. At least one amino acid sequence that is identical to any one of sequence numbers 258-261, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of. Therefore, in a preferred embodiment, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptide or protein of the TP53 tumor neoantigen. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 258-261, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0167] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 258-261, or C-terminal shortened versions of SEQ ID NOs. 258-261. In embodiments, the immunogenic fragments of SEQ ID NOs. 258-261 are shortened 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. 258-261 still contain at least one epitope capable of inducing an immune response, preferably at least one T-cell epitope.

[0168] (DIP2B, TRRAP, RAC1, AP4B1, VWA5A, CLTC, RPL9, RPAP1, RPE65, PATZ1, PIK3CA, NFE2L2, CNOT9 amino acid and nucleic acid sequences) In embodiments, at least one antigenic peptide is selected from DIP2B, TRRAP, RAC1, AP4B1, VWA5A, CLTC, RPL9, RPAP1, RPE65, PATZ1, PIK3CA, NFE2L2, or CNOT9 tumor neoantigen peptides or proteins. At least one amino acid sequence that is identical to any one of sequence numbers 287-317, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of. Therefore, in embodiments, the artificial nucleic acid includes at least one cds encoding at least one tumor antigen. The tumor antigen includes or consists of at least one antigenic peptide selected from the peptides or proteins of DIP2B, TRRAP, RAC1, AP4B1, VWA5A, CLTC, RPL9, RPAP1, RPE65, and PATZ1 tumor neoantigens. Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 287-317, or at least 80% identical to it, The immunogenic fragment or its variant, It includes or consists of.

[0169] In this context, the fragments may be N-terminal shortened versions of SEQ ID NOs. 287-317 or C-terminal shortened versions of SEQ ID NOs. 287-317. In embodiments, the immunogenic fragments of SEQ ID NOs. 287-317 are shortened 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. 287-317 still contain at least one epitope, preferably at least one T-cell epitope, capable of inducing an immune response.

[0170] The following describes preferred nucleic acid coding sequences that encode specific antigenic peptides of tumor neoantigens as defined herein. These sequences may be included within at least one coding sequence of the artificial nucleic acid of the present invention.

[0171] In a preferred embodiment, at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from a tumor neoantigen peptide or protein, wherein the nucleic acid sequence is It is identical to any one of sequence numbers 449-511, or is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments or any of those variants, Preferably, in this context, the fragment has a length of at least 24 nucleotides.

[0172] In a preferred embodiment, at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from a tumor neoantigen peptide or protein, wherein the nucleic acid sequence is It is identical to any one of sequence numbers 449-480, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments or any of their variants, Preferably, in this context, the fragment has a length of at least 24 nucleotides.

[0173] Preferably, the nucleic acid sequence encoding at least one antigenic peptide derived from the tumor neoantigen is a codon-modified cds. Preferably, the codon-modified cds are selected from C-maximized cds, CAI-maximized cds, cds adapted for human codon use, G / C content modified cds, and G / C optimized cds (all as defined herein), or any combination thereof. In a preferred embodiment in this context, at least one codon-modified cds is a G / C optimized cds.

[0174] Therefore, in a preferred embodiment, at least one coding sequence comprises a G / C optimized nucleic acid sequence encoding at least one antigenic peptide selected from a tumor neoantigen peptide or protein, wherein the nucleic acid sequence is It is identical to any one of sequence numbers 449-511, or is at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments, Preferably, in this context, the fragment has a length of at least 24 nucleotides.

[0175] Table 2 shows preferred amino acid and nucleic acid sequences in this context. Each row corresponds to the appropriate antigenic peptide of the tumor neoantigen. Column A provides a brief description of each tumor neoantigen. Column B shows the sequence number of the protein (amino acid sequence) in the preferred antigenic peptide of the tumor neoantigen. Column C shows the sequence number of the corresponding G / C optimized nucleic acid sequence encoding the preferred antigenic peptide of the tumor neoantigen. Column F indicates the main cancer species to which the antigen is applicable (N = non-small cell lung cancer; M = melanoma).

[0176] [Table 3]

[0177] In a particularly preferred embodiment, the antigenic peptide comprises or consists of at least one amino acid sequence selected from column B of Table 2, or an immunogenic fragment thereof. In a more preferred embodiment, the antigenic peptide comprises or consists of at least one amino acid sequence selected from column B, rows 1 to 11 of Table 2, or an immunogenic fragment thereof.

[0178] In a particularly preferred embodiment, the at least one coding sequence of the artificial nucleic acid of the present invention includes or consists of at least one nucleic acid sequence, as shown in column C of Table 2. In a further preferred embodiment, in a particularly preferred embodiment, the at least one coding sequence of the artificial nucleic acid of the present invention includes or consists of at least one nucleic acid sequence, as shown in column C, rows 1 to 11 of Table 2.

[0179] [Further characteristics of peptides or proteins] In embodiments, at least one coding sequence codes for 2 to 10 different tumor antigens as described herein, preferably 2 to 5 different tumor antigens as described herein. Thus, the coding sequence may code for multiple tumor antigens obtained in the form of a fusion construct.

[0180] Appropriately, each different tumor antigen is selected to enhance coverage in a specific cancerous disease (NSCLC, HNSCC, or melanoma). Appropriately, the coverage value for all different tumor antigens is 0.5 or higher, preferably 0.6 or higher, and more preferably 0.7 or higher.

[0181] In the context of this invention, "different tumor antigens" refers to differences at the amino acid sequence level and should be understood as differences in at least one amino acid position or amino acid sequence length. Therefore, "different tumor antigens" may originate from the same tumor neoantigen, but they must differ in at least one amino acid position or length.

[0182] Therefore, at least one coding sequence may encode 2 to 10 different tumor antigens, preferably 2 to 5 different tumor antigens. Here, the different tumor antigens are selected from peptides or proteins encoded by long non-coding RNA (lncRNA) as defined herein, and / or peptides or proteins of tumor neoantigens as defined herein, or immunogenic fragments of either thereof or variants thereof. Appropriate amino acid sequences and nucleic acid sequences in this context are defined above and may be selected from, in particular, Tables 1A and B and Table 2.

[0183] In preferred embodiments in this context, different tumor antigens are selected to cover different HLA types. Thus, different tumor antigens are selected from among the at least one tumor antigen as defined herein, including a tumor antigen comprising an epitope that binds to at least one HLA-A type, a tumor antigen comprising an epitope that binds to at least one HLA-B type, and a tumor antigen comprising an epitope that binds to at least one HLA-C type.

[0184] In embodiments, different tumor antigens are separated by linker elements. Therefore, the coding sequence may encode multiple different tumor antigens obtained in the form of a fusion construct, where the different tumor antigens within the fusion construct are separated by linker elements. Preferably, such linkers are non-immunogenic. Suitable linker elements may be selected from sequence numbers 2937, 76400-76418, 77018-77058 described in international publication application WO2019008001.

[0185] In embodiments, at least one cds encodes at least one additional amino acid sequence selected from at least one immune response activation signaling protein, preferably encoding at least one transmembrane domain and / or at least one cytoplasmic domain derived from the at least one immune response activation signaling protein.

[0186] Preferably, immune response-activating signaling proteins are located outside the cell membrane and are readily and repeatedly taken up into the endosomal pathway. This uptake is preferably for targeting the encoded and translated antigen to a desired intracellular pathway. This intracellular pathway preferably crosses with the MHC class I pathway and, in particular, the MHC class II pathway, thereby enabling effective delivery of the antigen sequence to the MHC class I and MHC class II processing compartments (as described in WO2019008001, all of which are incorporated by reference).

[0187] In a preferred embodiment, the immune response activation signaling protein is selected from CTLA4.

[0188] In a preferred embodiment, at least one immune response activation signaling protein is At least one amino acid sequence that is identical to any one of sequence numbers 76636 (described in International Publication Patent Application WO2019008001), or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of them, or Any of those fragments or any of those variants, It includes or consists of.

[0189] In embodiments, at least one coding sequence encodes at least one additional amino acid sequence selected from at least one T helper epitope. A suitable T helper epitope may be selected from sequence numbers 3038-3294 described in international published patent application WO2019008001.

[0190] In embodiments, 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 NOs. 1-156, 76948-76951 of International Publication Application WO2019008001. In preferred embodiments, at least one signal peptide is selected from CTLA4.

[0191] [Characteristics and Embodiments of Nucleic Acid Sequences] Appropriate features and embodiments of the artificial nucleic acid sequences of the present invention will be described in detail below (types of nucleic acids, structures of nucleic acids, elements of nucleic acids, modifications of nucleic acids, etc.). In particular, the features defining the nucleic acid sequences may also be applied to any nucleic acid sequences in any other aspect of the present invention (compositions, combinations, kits, medical applications, etc.).

[0192] Appropriate coding array: In a preferred embodiment, the artificial nucleic acid of the present invention comprises at least one coding sequence encoding at least one tumor antigen as defined herein. In this context, any coding sequence encoding at least one tumor antigen as defined herein, or a fragment thereof or a variant thereof, may be understood as a suitable coding sequence and therefore may be included in the nucleic acid of the present invention.

[0193] In a preferred embodiment, the artificial nucleic acid is a modified nucleic acid and / or a stabilized nucleic acid.

[0194] According to a preferred embodiment, the artificial nucleic acid may be provided as a “stabilized nucleic acid,” i.e., a nucleic acid with improved resistance to degradation in vivo, and / or a nucleic acid with improved stability in vivo, and / or a nucleic acid with improved translational ability in vivo. This is particularly important in embodiments where the nucleic acid is RNA.

[0195] Preferably, the artificial nucleic acid of the present invention may be provided as a "stabilized nucleic acid," preferably as a "stabilized RNA."

[0196] The following describes appropriate modifications / adaptations that can "stabilize" nucleic acids, preferably RNA.

[0197] In a particularly preferred embodiment, the artificial nucleic acid comprises at least one codon-modified cds. Preferably, the amino acid sequence encoded by 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.

[0198] The term "codon-modified coding sequence" refers to a coding sequence in which at least one codon (a chain of three nucleotides encoding one amino acid) differs from the corresponding wild-type or reference coding sequence. Appropriately, codon-modified cds in the context of the present invention may exhibit improved resistance to degradation in vivo, and / or improved stability in vivo, and / or improved translational activity in vivo. In its broadest sense, codon modification utilizes the degeneracy of the genetic code, where multiple codons may encode the same amino acid and may be used interchangeably to optimize / modify the coding sequence for in vivo applications.

[0199] In an embodiment, at least one cds is a codon-modified cds, where the codon-modified cds is a C-maximized cds (further defined in WO2021239880 [p.122, lines 33 to 39], incorporated herein by reference); a CAI-maximized cds (further defined in WO2021239880 [p.123, lines 33 to 44], incorporated herein by reference); a cds adapted to human codon usage (further defined in WO2021239880 [p.123, lines 7 to 17], incorporated herein by reference); a G / C content-modified cds (further defined in WO2021239880 [p.123, lines 19 to 31], incorporated herein by reference); a G / C-optimized cds; or a combination of any of these.

[0200] When transfected into a mammalian host cell, a nucleic acid comprising a codon-modified coding sequence has stability for between 12 and 18 hours, or for 18 hours or more (such as 24, 36, 48, 60, 72 hours, etc.), or for 72 hours or more, and can be expressed by the mammalian host cell.

[0201] When transfected into a mammalian host cell, a nucleic acid comprising a codon-modified coding sequence is translated into a protein, where the amount of protein is at least equivalent, or preferably, at least 10% or more, at least 20% or more, at least 30% or more, at least 40% or more, at least 50% or more, at least 100% or more, or at least 200% or more relative to the amount of protein obtained by transfecting the mammalian host cell with a native or wild-type or reference coding sequence.

[0202] In particularly preferred embodiments, the artificial nucleic acid may be modified, where the G / C content of at least one coding sequence may be optimized compared to the G / C content of the corresponding wild-type or reference coding sequence (“G / C-optimized coding sequence”). “Optimization” in this context refers to a coding sequence in which the G / C content has been increased to the highest possible value substantially. Generation of G / C-content-optimized nucleic acid sequences may be carried out using the methods described in WO2002098443. In this context, the disclosure of WO2002098443 is incorporated in its entirety into the present invention.

[0203] In preferred embodiments, at least one codon-modified coding sequence is a G / C-optimized coding sequence. Such G / C-optimized nucleic acid sequences are shown in Table 1A or 1B, or Table 2.

[0204] In preferred embodiments, at least one cds of the artificial nucleic acid, preferably RNA, has a G / C content of at least about 50%, 55%, or 60%. In certain embodiments, at least one cds 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 particularly preferred embodiments, the artificial nucleic acid may be modified, where the G / C content of at least one cds may be optimized compared to the G / C content of the corresponding wild-type or reference cds (“G / C-optimized coding sequence”). “Optimization” in this context refers to a coding sequence in which the G / C content has been increased to the highest possible value substantially. Generation of G / C-content-optimized nucleic acid sequences may be carried out using the methods described in WO2002098443. In this context, the disclosure of WO2002098443 is incorporated in its entirety into the present invention.

[0205] UTRs: In a preferred embodiment, the artificial nucleic acid (preferably RNA) of the present invention comprises at least one untranslated region (UTR) or UTR element.

[0206] The term "untranslated region," "UTR," or "UTR element" refers to a portion of a nucleic acid molecule typically located at the 5' or 3' end of a coding sequence. UTRs are not translated into proteins. UTRs may be part of a nucleic acid (e.g., RNA). UTRs may contain elements that regulate gene expression (also called regulatory elements). Such regulatory elements may include, for example, ribosome binding sites, miRNA binding sites, or promoter elements.

[0207] In a preferred embodiment, the artificial nucleic acid comprises a coding sequence, as well as a 5'-UTR and / or 3'-UTR. In particular, the UTR may have regulatory sequence elements that determine RNA turnover, stability, and localization. Furthermore, the UTR may have sequence elements that facilitate translation. In medical applications, the translation of the nucleic acid into at least one peptide or protein is critical to therapeutic efficacy. Certain combinations of 3'-UTR and / or 5'-UTR may facilitate the expression of an operablely linked coding sequence encoding a peptide or protein as defined herein. Nucleic acid molecules having the aforementioned combinations of UTRs advantageously enable rapid and transient expression of the encoded tumor antigen after administration to a subject.

[0208] Suitablely, 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 3'-UTR may be derived from a naturally occurring gene or may be synthetically designed. In a preferred embodiment, the artificial nucleic acid comprises at least one coding sequence as defined herein, which is operably linked to at least one 3'-UTR and / or at least one 5'-UTR.

[0209] Preferably, at least one untranslated region (UTR) is selected from at least one heterogeneous 5'-UTR and / or at least one heterogeneous 3'-UTR.

[0210] As used herein, the term “heterogeneous” sequence or UTR is intended to mean a nucleic acid sequence or UTR that does not originate from the same gene, the same genome fusion, or the same naturally occurring transcript (e.g., lncRNA). Therefore, heterogeneous sequences or UTRs may originate from the same organism (e.g., humans) or different organisms. Heterogeneous sequences or UTRs do not naturally occur within the same nucleic acid.

[0211] In a preferred embodiment, the artificial nucleic acid of the present invention comprises at least one 3'-UTR.

[0212] The term "3'-untranslated region" or "3'-UTR" refers to a portion of nucleic acid located at the 3' end (i.e., downstream) of a cds sequence that is not translated into protein. The 3'-UTR may also be a portion of nucleic acid located between the cds sequence and any terminal poly(A) sequence. The 3'-UTR may contain elements (also called regulatory elements) that control gene expression. Such regulatory elements may be, for example, ribosome binding sites or miRNA binding sites.

[0213] Preferably, the artificial nucleic acid comprises at least one 3'-UTR, which may be derived from a gene associated with RNA with an extended half-life (i.e., stable RNA provision).

[0214] In some embodiments, the 3'-UTR includes one or more polyadenylation signals, protein binding sites that affect nucleic acid stability in intracellular localization, or one or more miRNAs or miRNA binding sites.

[0215] In a preferred embodiment, the artificial nucleic acid comprises at least one 3'-UTR, where the at least one 3'-UTR is PSMB3, ALB7, α-globin, β-globin, ANXA4, CASP1, COX6B1, FIG4, GNAS, NDUFA1, RPS9, SLC7A3, or TUBB4B, or Any one homolog, fragment, or variant of these genes, It contains, or consists of, a nucleic acid sequence derived from, or selected from, the 3'-UTR of a gene selected from.

[0216] 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 is Nucleic acid sequences that are identical to any one of sequence numbers 66-95 or 112-123, or that are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Those fragments or their variants, It includes or consists of.

[0217] In other embodiments, at least one 3'-UTR is Nucleic acid sequences that are identical to any one of sequence numbers 96-111, or that are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments or any of those variants, It includes or consists of.

[0218] In a particularly preferred embodiment, the artificial nucleic acid comprises a 3'-UTR derived from or selected from the PSMB3 gene, wherein at least one heterologous 3'-UTR is A nucleic acid sequence that is identical to SEQ ID NOs: 66, 67, 112 - 123 or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to them, preferably the nucleic acid sequence of SEQ ID NO: 67, or any fragment thereof or any variant thereof, comprising or consisting of.

[0219] In a preferred embodiment, the artificial nucleic acid of the present invention comprises at least one 5'-UTR.

[0220] The term "5'-untranslated region" or "5'-UTR" refers to a part of the nucleic acid located on the 5' side (i.e., "upstream") of the cds that is not translated into protein. The 5'-UTR may be a part of the nucleic acid located on the 5' side of the cds. Usually, the 5'-UTR starts from the transcription start site and ends before the start codon of the cds. The 5'-UTR may contain elements (also called regulatory elements) that control gene expression. Such regulatory elements may be, for example, ribosome binding sites, miRNA binding sites, etc. The 5'-UTR may be modified, for example, an enzymatic or co-transcriptional 5'-cap structure may be added (such as in the case of mRNA described later).

[0221] Preferably, the artificial nucleic acid comprises at least one 5'-UTR, and the 5'-UTR may be derived from a gene related to RNA with an extended half-life (i.e., providing stable RNA).

[0222] In some embodiments, the 5'-UTR contains one or more protein binding sites that affect the intracellular localization of nucleic acid stability, or one or more miRNAs or miRNA binding sites (as defined above).

[0223] Therefore, the miRNA or miRNA-binding site defined above may be removed from or introduced into the 5'-UTR. This is to tailor nucleic acid expression to the desired cell type or tissue (such as muscle cells).

[0224] In a preferred embodiment, the artificial nucleic acid comprises at least one 5'-UTR, where the at least one 5'-UTR is HSD17B4, RPL32, AIG1, α-globin, ASAH1, ATP5A1, COX6C, DPYSL2, MDR, MP68, NDUFA4, NOSIP, RPL31, RPL35A, SLC7A3, TUBB4B, or UBQLN, or Any one homolog, fragment, or variant of these genes, It contains nucleic acid sequences derived from, or selected from, the 5'-UTR of a gene selected from.

[0225] 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 UBQLN is Nucleic acid sequences that are identical to any one of sequence numbers 12-45, 64, or 65, or that are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments or any of those variants, It includes or consists of.

[0226] In other embodiments, at least one 5'-UTR is Nucleic acid sequences that are identical to any one of sequence numbers 46-63, or are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those fragments or any of those variants, It includes or consists of.

[0227] In a particularly preferred embodiment, the artificial nucleic acid comprises a 5'-UTR derived from or selected from the HSD17B4 gene, wherein at least one heterologous 5'-UTR is Nucleic acid sequences identical to sequence numbers 12, 13, 64, 65, preferably sequence number 13, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, It includes or consists of.

[0228] In embodiments, at least one nucleic acid (preferably RNA) comprises at least one cds, the cds being operably linked to a 3'-UTR element and / or a 5'-UTR element selected from a 5'-UTR / 3'-UTR combination (5'UTR / 3'UTR) disclosed in WO2021239880 [p.127, line 35 to p.128, line 2] and incorporated herein by reference.

[0229] In a preferred embodiment, at least one heterogeneous 5'-UTR is selected from HSD17B4 and at least one heterogeneous 3'-UTR is selected from PSMB3.

[0230] Accordingly, in a particularly preferred embodiment, the artificial nucleic acid (preferably RNA) comprises at least one cds as defined herein, encoding at least one tumor antigen as defined herein, wherein the cds are operably linked to HSD17B4 5'-UTR and PSMB3 3'-UTR (HSD17B4 / PSMB3(a-1)). The inventors have shown that this embodiment is particularly beneficial for the expression of tumor antigens in human cells.

[0231] In various embodiments, nucleic acids (e.g., RNA) are monocistronic, bicistronic, or multicistronic.

[0232] In a preferred embodiment, the nucleic acid (preferably RNA) is monocistronic.

[0233] Therefore, in a particularly preferred embodiment, the artificial nucleic acid includes a ribosome-binding site (also called a "Kozak sequence"), The ribosome binding site in question is Sequence ID 1 or 2, or any one of the sequences GCCGCCACC(DNA), GCCGCCACC(RNA), GCCACC(DNA), GCCACC(RNA), ACC(DNA), or ACC(RNA), or a sequence that is identical to at least 80%, 85%, 90%, or 95% identical to any one of these, or It is any fragment or any variant thereof. In a preferred embodiment, the “Kozak sequence” comprises or consists of the RNA sequence ACC.

[0234] DNA and RNA constructs: In a particularly preferred embodiment, the artificial nucleic acid of the present invention is an isolated nucleic acid. The term “isolated nucleic acid” refers to an isolated molecule or an aggregate of isolated molecules, but does not include the cell or object containing the nucleic acid. For example, “isolated nucleic acid” may be an artificial nucleic acid isolated or purified from a cell (cell culture, bacterial culture, etc.), or an artificial nucleic acid (e.g., RNA) isolated from RNA in in vitro transcription.

[0235] In a particularly preferred embodiment, the artificial nucleic acid of the present invention is a therapeutic nucleic acid.

[0236] Therefore, artificial nucleic acids are appropriately used in a therapeutic context, and in particular, are used to provide a therapeutic modality that provides at least one tumor antigen according to the present invention.

[0237] In embodiments, the artificial nucleic acid of the present invention is selected from DNA or RNA.

[0238] In embodiments, the artificial nucleic acid of the present invention is selected from DNA. The DNA may be any type of DNA (including single-stranded DNA, double-stranded DNA, linear DNA, and circular DNA) including cds as defined herein. Suitable DNA in the context of the present invention may be selected from bacterial plasmids, adenoviruses, poxviruses, parapoxviruses (Aarf viruses), vaccinia viruses, avianpox viruses, herpesviruses, adeno-associated viruses (AAVs), alphaviruses, lentiviruses, lambda phages, lymphocytic choriomeningitis viruses, Listeria species, and Salmonella species. In preferred embodiments, the DNA is viral DNA, preferably AAV DNA.

[0239] In a particularly preferred embodiment, the artificial nucleic acid of the present invention is RNA. The RNA may be any type of RNA (including single-stranded RNA, double-stranded RNA, linear RNA, and circular RNA) that includes cds as defined herein. In a preferred embodiment, the RNA may be any type of RNA (including single-stranded RNA, double-stranded RNA, linear RNA, and circular RNA) that includes cds as defined herein, wherein the 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, and is preferably selected from mRNA or circular RNA.

[0240] In embodiments, the RNA is not an lncRNA. In particular, the artificial nucleic acid of the present invention is not an lncRNA 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. This means that the RNA of the present invention may, of course, include a coding sequence encoding a peptide 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, but the RNA of the present invention is not identical in sequence and length to 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.

[0241] In embodiments, the RNA is circular RNA. A "circular RNA" (circRNA) is RNA linked in a circular manner and therefore does not contain a 3' or 5' end. The circRNA comprises at least one cds as defined herein. Designs of the circRNA construct can be incorporated herein by reference from claims 1 to 51 of WO2023073228.

[0242] In other embodiments, the RNA is replicon RNA or self-replicating RNA. Such a construct may encode, for example, a replicase element derived from an alphavirus (SFV, SIN, VEE, or RRV, etc.) and cds as defined herein.

[0243] In a particularly preferred embodiment, the RNA is selected from mRNA. Therefore, the artificial nucleic acid of the present invention is mRNA, and appropriately isolated mRNA.

[0244] In the context of the present invention, mRNA is preferred for providing the tumor antigen according to the present invention. This is because mRNA can be administered in controlled doses, exhibits transient expression, is completely degraded after protein synthesis, and does not carry the risk of insertional mutations.

[0245] Preferably, the artificial nucleic acid (preferably RNA) has about 50 to about 20,000 nucleotides, or It contains approximately 500 to approximately 10,000 nucleotides, or approximately 1,000 to approximately 10,000 nucleotides, or preferably approximately 1,000 to approximately 5,000 nucleotides, or approximately 2,000 to approximately 5,000 nucleotides.

[0246] Poly(N) sequence, histone stem loop: In a preferred embodiment, the artificial nucleic acid comprises at least one poly(N) sequence, for example, at least one poly(A) sequence, at least one poly(U) sequence, at least one poly(C) sequence, or a combination thereof.

[0247] In a preferred embodiment, the artificial nucleic acid (e.g., RNA) comprises at least one poly(A) sequence. In some embodiments, the artificial nucleic acid comprises at least two, three, or more poly(A) sequences.

[0248] The term "poly(A) sequence" or "poly(A) tail" refers to an adenosine sequence of up to 1000 nucleotides typically located at the 3' end of linear RNA. Poly(A) sequences are usually homopolymerized. Alternatively, a poly(A) sequence may be interrupted by at least one nucleotide distinct from adenosine.

[0249] In embodiments, at least one poly(A) sequence may contain 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. Appropriately, the length of the poly(A) sequence may be at least about 50, 64, 75, 100, 200, 300, 400, or 500 adenosine nucleotides, or more, and may preferably be consecutive adenosine nucleotides.

[0250] In a particularly preferred embodiment, at least one poly(A) sequence contains about 100 adenosine nucleotides (A100) and about 100 consecutive adenosine nucleotides.

[0251] In another embodiment, at least one nucleic acid comprises at least one interrupted poly(A) sequence, where the poly(A) sequence is interrupted by non-adenosine nucleotides, preferably about 10 non-adenosine (N10) nucleotides. In this context, the poly(A) sequence A30-N10-A70 is preferred.

[0252] The poly(A) sequences as defined herein may 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 the 3' terminal A nucleotide of at least one poly(A) sequence. The term “located directly at the 3' end” means strictly located at the 3' end; in other words, the 3' end of the nucleic acid should be understood to consist of a poly(A) sequence ending in A.

[0253] Advantageously, because the 3' end of the RNA of the present invention terminates with an adenosine nucleotide, when administered to humans, for example, as a pharmaceutical, it reduces the induction of interferon (such as IFNα). This is important because the induction of interferon (such as IFNα) is considered to be one of the main causes of side effects.

[0254] Therefore, in a particularly preferred embodiment, the artificial nucleic acid (e.g., RNA) of the present invention comprises a poly(A) sequence of about 100 consecutive adenosine nucleotides. This poly(A) sequence is located directly at the 3' end of the RNA, and optionally, the 3' terminal nucleotide is adenosine.

[0255] In a preferred embodiment, the artificial nucleic acid (e.g., RNA) comprises at least one histone stem-loop (hSL) or histone stem-loop structure. In the context of the present 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, primarily found in histone mRNA. The histone stem-loop sequence / structure may be appropriately selected from the hSL sequences disclosed in WO2012019780. The hSL sequences that may be used herein may be derived from formula (I) or (II) of WO2012019780. Thus, the artificial nucleic acid may comprise at least one hSL sequence derived from a specific formula (Ia) or (IIa) of WO2012019780.

[0256] In a preferred embodiment, at least one histone stem-loop array is A nucleic acid sequence that is identical to either sequence number 3 or 4, or is at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these sequences, or Any of those fragments or any of those variants, It includes or consists of the following. Preferably, the histone stem-loop sequence includes or consists of the nucleic acid sequence shown in Sequence ID No. 4, or a fragment or variant thereof.

[0257] In a preferred embodiment, the artificial nucleic acid includes a 3' terminal sequence element, which represents the 3' end of RNA. The 3' terminal sequence element may include at least one poly(N) sequence as defined herein, and optionally, at least one hSL as defined herein.

[0258] In a preferred embodiment, the artificial nucleic acid comprises at least one 3' terminal sequence element, the sequence element being RNA sequences that are identical to any one of sequence numbers 5-11, or that are at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Fragments of those sequences or variants of those sequences, It includes or consists of.

[0259] In a preferred embodiment, the artificial nucleic acid comprises a 3' terminal sequence element (including an hSL as defined herein) followed by a poly(A) sequence (containing about 100 consecutive adenosines).

[0260] In a particularly preferred embodiment, the artificial nucleic acid includes a 3' terminal sequence element, and the sequence element is A nucleic acid sequence that is identical to sequence number 5 or 6, or identical to it by at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or The fragment or variant thereof, It includes or consists of.

[0261] In some embodiments, the artificial nucleic acid includes a 5' terminal sequence element, and the sequence element is Nucleic acid sequences that are identical to any one of the sequences AGGAGA, GGGAGA, GGGAAA, AGAAUA, AGAUUA, GAUGGG, or GGGCG, or that are at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Fragments of those sequences or variants of those sequences, preferably AGGAGA, It includes or consists of such a 5' terminal sequence element, for example, a binding site for T7 RNA polymerase. Furthermore, the first nucleotide of the 5' terminal start sequence may preferably include a 2'O methylation (such as 2'O methylated guanosine or 2'O methylated adenosine).

[0262] Modified nucleotides: In various embodiments, the artificial nucleic acid (preferably RNA) is modified, where modification refers to chemical modifications including sugar modification or base modification in addition to skeletal modification.

[0263] Modified nucleic acids or RNA may include nucleotide analogs / modifiers (such as skeletal modifications, sugar modifications, or base modifications). Skeletal modification in the context of the present invention refers to modifications in which the phosphate groups of the RNA nucleotide skeleton are chemically modified. Sugar modification in the context of the present invention refers to chemical modifications of sugars in the nucleotides of RNA. Furthermore, base modification in the context of the present invention refers to chemical modifications of base moieties in the nucleotides of RNA. In this context, the nucleotide analogs or modifiers are preferably selected from nucleotide analogs applicable to transcription and / or translation.

[0264] Therefore, in a preferred embodiment, the nucleic acid (preferably the RNA of the present invention) comprises at least one modified nucleotide.

[0265] In some embodiments, at least one modified nucleotide is selected from pseudouridine, N1-methylpseudridine, N1-ethylpseudridine, 2-thiouridine, 4'-thiouridine, 5-methylcytosine, 5-methyluridine, 2-thio-1-methyl-1-deaza-pseudridine, 2-thio-1-methylpseudridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudridine, 2-thio-dihydrouridine, 2-thiopseudridine, 4-methoxy-2-thiopseudridine, 4-methoxypseudridine, 4-thio-1-methylpseudridine, 4-thiopseudridine, 5-aza-uridine, dihydropseudridine, 5-methoxyuridine, and 2'-O-methyluridine.

[0266] In this context, pseudouridine (ψ) or N1-methylpseudridine (m1ψ) are particularly preferred. Therefore, in a preferred embodiment, the nucleic acid is RNA comprising at least one modified nucleotide, preferably a modified nucleotide selected from pseudouridine (ψ) or N1-methylpseudridine (m1ψ).

[0267] In some embodiments, substantially all (e.g., substantially 100%) of the uracil in the coding sequence (or whole nucleic acid sequence) has a chemical modification, preferably a chemical modification at the 5-position of uracil.

[0268] In a preferred embodiment, 100% of the uracil in the entire nucleic acid sequence (preferably an RNA sequence) is replaced by a modified nucleotide (preferably N1-methylpseudridine (m1ψ)). Alternatively, 100% of the uracil in the entire nucleic acid sequence (preferably an RNA sequence) is replaced by pseudouridine (ψ).

[0269] In preferred embodiments, the artificial nucleic acid (preferably RNA) does not contain chemically modified nucleotides. In particular, the 5'-cap structure described later is not typically considered a chemically modified nucleotide. Therefore, the artificial nucleic acid (preferably RNA) contains a sequence consisting only of G, C, A, and U nucleotides, and thus does not contain modified nucleotides, and optionally contains a 5'-cap structure. RNA constructs that do not contain chemically modified nucleotides may be beneficial in the context of cancer therapy. This is because a stronger T cell response can be induced by expressing cancer antigens via unmodified RNA (compared to RNA modified with m1ψ or ψ).

[0270] In preferred embodiments, the artificial nucleic acid (preferably the RNA of the present invention) does not contain sites substituted with N1-methylpseudridine (m1ψ) or sites substituted with pseudouridine (ψ).

[0271] Cap structure: In a preferred embodiment, the artificial nucleic acid is RNA containing a 5'-cap structure.

[0272] Such 5'-cap structures appropriately stabilize nucleic acids and / or enhance the expression of encoded tumor antigens and / or reduce the stimulation of the innate immune system after administration.

[0273] Therefore, in a preferred embodiment, the artificial nucleic acid (preferably RNA) includes a 5'-cap structure (preferably m7G, cap0, cap1, cap2, modified cap0 structure, or modified cap1 structure).

[0274] As used herein, the term "5'-cap structure" is intended to be as recognized and understood by those skilled in the art, and refers, for example, to a 5'-modified nucleotide located at the 5' end of RNA, particularly a guanine nucleotide. Preferably, the 5'-cap structure is linked to the RNA via a 5'-5'-triphosphate bond.

[0275] Possibly suitable 5'-cap structures in the context of the present invention include cap0 (methylation of the first base, e.g., m7GpppN), cap1 (additional methylation of ribose of the nucleotide adjacent to m7GpppN), cap2 (additional methylation of ribose of the second downstream nucleotide of m7GpppN), cap3 (additional methylation of ribose of the third downstream nucleotide of m7GpppN), cap4 (additional methylation of ribose of the fourth downstream nucleotide of m7GpppN), ARCA (anti-reverse cap analogue), modified ARCA (e.g., phosphothioate modified ARCA), inosine, N1-methyl-guanosine, 2'-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

[0276] Appropriately, a 5'-cap (cap0 or cap1) structure may be formed using a cap analog during chemical RNA synthesis or in vitro RNA transcription (cotranscriptional capping).

[0277] As used herein, the term “cap analog” is intended to be as recognized and understood by those skilled in the art, and refers to a non-polymerizable dinucleotide or trinucleotide that, when incorporated into the 5' end of a nucleic acid molecule, has a capping function that promotes translation or localization and / or prevents the degradation of the RNA molecule. “Non-polymerizable” means that the cap analog is incorporated only into the 5' end. This is because the cap analog does not have a 5' triphosphate and is therefore not extended in the 3' direction by template-dependent polymerases (particularly template-dependent RNA polymerases).

[0278] In embodiments, the cap1 structure is produced 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 may be used as appropriate to co-transfer and produce the cap1 structure. Furthermore, any cap structure defined in claims 1 to 37 of WO2023007019 may be used as appropriate to produce cap1.

[0279] In a preferred embodiment, the 5' cap structure may be appropriately added co-transcribed using a trinucleotide cap analog as defined herein, preferably in an in vitro transcription reaction of RNA as defined herein.

[0280] In a particularly preferred embodiment, the artificial nucleic acid (preferably the RNA of the present invention) includes a cap1 structure or a modified cap1 structure.

[0281] In a preferred embodiment, the cap1 structure is formed via co-transcriptional capping using the trinucleotide cap analog m7G(5')ppp(5')(2'OMeA)pG or m7G(5')ppp(5')(2'OMeG)pG. In this context, the cap1 analog is particularly preferred to be m7G(5')ppp(5')(2'OMeA)pG.

[0282] In another preferred embodiment, the cap1 structure is a modified cap1 structure, formed by co-transcriptional capping using the trinucleotide cap analog 3'OMe-m7G(5')ppp(5')(2'OMeA)pG.

[0283] Alternatively, the 5'-cap structure may be formed via enzymatic capping using a capping enzyme (e.g., vaccinia virus capping enzyme and / or 2'-O methyltransferase) to generate cap0, cap1, or cap2 structures.

[0284] It is preferable that at least 80%, 85%, 90%, and 95% of the RNA species have a cap structure (preferably a cap1 structure), as determined by a capping assay (for example, the assay described in claims 27 to 46 of WO2015101416).

[0285] Further RNA characteristics: In various embodiments, the artificial nucleic acid is an RNA that provides at least one coding sequence encoding at least one tumor antigen as defined herein, which is produced after administration to cells or a subject.

[0286] Suitable elements preferably included in the RNA of the present invention include, for example, a 5' cap structure, a 5'UTR, a 3'UTR, an hSL, a poly(A) sequence (all as defined herein), and any chemical modifications as defined herein.

[0287] In a preferred embodiment, the RNA is preferably in vitro transcribed RNA (e.g., in vitro transcribed mRNA).

[0288] In a preferred embodiment, the nucleotide mixture used in 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. Thus, in a preferred embodiment, the nucleic acid of the present invention is an in vitro transcribed RNA, preferably where the in vitro transcription of RNA is carried out in the presence of a sequence-optimized nucleotide mixture.

[0289] In a preferred embodiment, the artificial nucleic acid of the present invention is purified RNA, preferably purified mRNA. Preferably, the RNA of the present invention is purified by at least one purification step.

[0290] The term "purified RNA" refers to RNA that has a higher purity than the raw material (e.g., crude IVT RNA) after a specific purification process. Typical impurities include peptides, proteins, spermidine, BSA, incomplete RNA fragments, dsRNA, free nucleotides, DNA, and buffer components. In this regard, it is desirable that the "purity of RNA" be as close to 100% as possible. Preferably, the "purified RNA" used herein has a purity of 75%, 80%, 85%, 90% or higher, or 95% or higher. Purity may be measured by analytical HPLC.

[0291] In embodiments, the nucleic acid is purified RNA purified by at least one of the following steps: (RP)-HPLC, AEX, size exclusion chromatography (SEC), hydroxyapatite chromatography, tangential flow filtration (TFF), filtration, precipitation, core bead flow-through chromatography, oligo(dT) purification, cellulosic purification, or any combination thereof. Preferably, the RNA is purified using RP-HPLC (preferably as described in WO2008077592) and / or TFF (preferably as described in WO2016193206) and / or oligo d(T) purification (preferably as described in WO2016180430) to remove, for example, double-stranded RNA, uncapped RNA, and / or RNA fragments.

[0292] In embodiments, the RNA has at least 60%, 70%, 80%, or 90% integrity. The term “RNA integrity” generally refers to the presence of a complete RNA sequence. RNA integrity may be determined by RP-HPLC and may be based on the measurement of the area under the peak of the expected full-length RNA in the chromatogram. In this regard, it is desirable that the “RNA integrity” is as close to 100% as possible.

[0293] In preferred embodiments, nucleic acids (preferably RNA) are suitable for use in the treatment or prevention of disease, disorder, or condition (preferably a disease, disorder, or condition of tumor or cancer, or any disease, disorder, or condition associated with tumor or cancer). The term “cancer” as used in this disclosure includes metastatic cancer.

[0294] Preferred nucleic acid construct: In various embodiments, the artificial nucleic acid includes at least the following elements: A) 5'-cap structure, preferably a 5'-cap structure as defined herein; B) At least one cds encoding at least one tumor antigen as defined herein; C) 5'-UTR and / or 3'-UTR, preferably 5'-UTR and / or 3'-UTR as defined herein; D) At least one poly(A) sequence, preferably as described herein.

[0295] In a preferred embodiment, the artificial nucleic acid (preferably RNA) preferably includes the following sequence elements in the 5' to 3' direction: A) 5'-cap structure, preferably a single-cap structure; B) Preferably a 5'-UTR selected from or derived from the HSD17B4 gene, or a fragment thereof; C) A coding sequence encoding at least one tumor antigen as defined herein; D) Preferably, a 3'-UTR selected from or derived from the PSMB3 gene, or a fragment thereof; E) Optionally, histone stem loops; and, F) A poly(A) sequence preferably containing about 100 A nucleotides.

[0296] In a preferred embodiment, the artificial nucleic acid (preferably RNA) preferably includes the following sequence elements in the 5' to 3' direction: A) 5'-cap structure, preferably a single-cap structure; B) Preferably a 5'-UTR selected from or derived from the HSD17B4 gene, or a fragment thereof; C) CDs encoding at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA), At least one amino acid sequence that is identical to any one of sequence numbers 124-146, 546-571, 756, or 757, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, CDs that include or consist of; D) Preferably, a 3'-UTR selected from or derived from the PSMB3 gene, or a fragment thereof; E) Optionally, histone stem loops; and, F) A poly(A) sequence preferably containing about 100 A nucleotides.

[0297] In a particularly preferred embodiment, the artificial nucleic acid is mRNA containing the following sequence elements in the 5' to 3' direction: A) 5'-cap structure, preferably a single-cap structure; B) A nucleic acid sequence that is identical to sequence number 13, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, A 5'-UTR that includes or consists of; C) CDs comprising a nucleic acid sequence encoding at least one antigenic peptide selected from or derived from a peptide or protein encoded by a long non-coding RNA (lncRNA), wherein the nucleic acid sequence is Nucleic acid sequences that are identical to any one of sequence numbers 318-448, 651-755, or 764-771, or that are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or cds; which are any fragment or any variant thereof. D) A nucleic acid sequence that is identical to sequence number 67, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or The fragment or variant thereof, 3'-UTR containing or consisting of; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides.

[0298] In a particularly preferred embodiment, the artificial nucleic acid is mRNA containing the following sequence elements in the 5' to 3' direction: A) 5'-cap structure, preferably a single-cap structure; B) A nucleic acid sequence that is identical to sequence number 13, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, A 5'-UTR that includes or consists of; C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from or derived from a neoantigen peptide or protein as defined herein, wherein the nucleic acid sequence is Nucleic acid sequences that are identical to any one of sequence numbers 449-511, or are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; D) A nucleic acid sequence that is identical to sequence number 67, or identical to it by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, 3'-UTR containing or consisting of; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides.

[0299] 2: A pharmaceutical composition containing at least one nucleic acid encoding a tumor antigen: In a second embodiment, 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 embodiment.

[0300] In particular, the features and embodiments described in the context of the first embodiment (the nucleic acid of the present invention) should be read and understood as appropriate embodiments of the pharmaceutical composition of the second embodiment, and vice versa.

[0301] In the context of the present invention, “composition” means any type of composition which may incorporate a specific component (e.g., a nucleic acid encoding at least one tumor antigen), and which may optionally incorporate further components, usually at least one pharmaceutically acceptable carrier or excipient. The composition may be a dry composition (such as a powder, granules, or solid freeze-dried form). Alternatively, the composition may be in liquid form, and each component may be incorporated independently in dissolved or dispersed form.

[0302] Preferably, at least one artificial nucleic acid in the pharmaceutical composition is selected from RNA as further defined in the first embodiment. In a particularly preferred embodiment, at least one nucleic acid in the pharmaceutical composition is selected from mRNA as further defined in the first embodiment.

[0303] In a preferred embodiment, the pharmaceutical composition comprises a plurality or at least one nucleic acid species (e.g., RNA species), preferably each nucleic acid species encoding a different tumor antigen.

[0304] Preferably, the pharmaceutical composition as defined herein may contain 2 to 10 nucleic acid species as defined in the first embodiment, and preferably 2 to 5 nucleic acid species as defined in the first embodiment. Here, each of the 2 to 10 nucleic acid species, preferably 2 to 5 nucleic acid species, encodes at least one different tumor antigen.

[0305] Appropriately, each nucleoside encoding a different tumor antigen is selected to enhance coverage in a specific cancerous disease (NSCLC, HNSCC, or melanoma). Appropriately, the coverage value for all different tumor antigens is 0.5 or higher, preferably 0.6 or higher, and more preferably 0.7 or higher.

[0306] In a preferred embodiment in this context, different tumor antigens (obtained by nucleic acid sequences) are selected to cover different HLA types. Thus, different tumor antigens are selected from among the at least one tumor antigen as defined herein, including a tumor antigen comprising an epitope that binds to at least one HLA-A type, a tumor antigen comprising an epitope that binds to at least one HLA-B type, and a tumor antigen comprising an epitope that binds to at least one HLA-C type.

[0307] In various embodiments, at least one artificial nucleic acid, preferably at least one RNA in the pharmaceutical composition, is formulated with a pharmaceutically acceptable carrier or excipient.

[0308] Formulation / compounding: In a preferred embodiment, at least one nucleic acid (preferably at least one RNA) forms a complex with or binds to at least one further compound to obtain a formulated composition. The formulation in this context may function as a transfection agent. The formulation in this context may also have a function of protecting the nucleic acid from degradation to enable storage, transport, etc.

[0309] In the embodiment, at least one nucleic acid (preferably at least one RNA) is formulated with at least one compound (such as a peptide, protein, lipid, polysaccharide, and / or polymer).

[0310] In embodiments, at least one artificial nucleic acid (preferably at least one RNA) is formulated with at least one cationic (cationic or preferably ionizable) compound, or a polycationic (cationic or preferably ionizable) compound. In preferred embodiments, at least one artificial nucleic acid (preferably at least one RNA) forms a complex with or binds to one or more cationic (cationic or preferably ionizable) compounds or polycationic compounds, or at least partially forms a complex with or partially binds to them.

[0311] The term "cationic or polycationic compound" refers to a molecule that is positively charged at pH values ​​in the 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 (e.g., pH values ​​in the range of about 7.2 to 7.5). Therefore, cationic components (cationic peptides, cationic proteins, cationic polymers, cationic polysaccharides, cationic lipids, etc.) may be any positively charged compound or polymer that is positively charged under physiological conditions. "Cationic or polycationic peptides or proteins" may contain at least one positively charged amino acid, or multiple positively charged amino acids (selected from Arg, His, Lys, or Orn, etc.). Therefore, "polycationic" components also fall within a range of multiple positive charges under given conditions.

[0312] In a preferred embodiment, at least one cationic or polycationic compound is selected from cationic or polycationic polymers, cationic or polycationic polysaccharides, cationic or polycationic lipids, cationic or polycationic proteins, cationic or polycationic peptides, or any combination thereof.

[0313] In a particularly preferred embodiment, at least one artificial nucleic acid, preferably at least one RNA in the pharmaceutical composition, is formulated within a lipid-based carrier.

[0314] In the context of the present invention, the term “lipid carrier” encompasses a lipid-based delivery system for nucleic acids (e.g., RNA) that includes lipid components. The lipid carrier may further include other components suitable for encapsulating / integrating / complexing nucleic acids (e.g., RNA). The nucleic acids may include cationic or polycationic polymers, cationic or polycationic polysaccharides, cationic or polycationic proteins, cationic or polycationic peptides, or any combination thereof.

[0315] At least one nucleic acid (preferably RNA) may be fully or partially incorporated into or encapsulated within a lipid carrier. Here, the at least one nucleic acid may be located in relation to the internal space of the lipid carrier, within the lipid layer / membrane of the lipid carrier, or outside the lipid carrier. Incorporating a nucleic acid into a lipid carrier may be referred to as "encapsulation."

[0316] The term “encapsulated” (including incorporated, complexed, encapsulated, partially encapsulated, bound, partially bound, etc.) refers to the intrinsic and stable combination of nucleic acids (preferably RNA) with one or more lipids within a lipid carrier (such as a larger complex or aggregate), preferably without covalent bonding. Nucleic acids (e.g., RNA) encapsulated in a lipid carrier may be entirely or partially present within the lipid carrier (e.g., the lipid portion and / or internal space) and / or within the lipid layer / membrane of the lipid carrier. In this specification, encapsulating nucleic acids (e.g., RNA) in a lipid carrier is also referred to as “incorporated” because the nucleic acids (e.g., RNA) are preferably contained within the lipid carrier. While not intended to be theoretically bound, the purpose of incorporating or encapsulating nucleic acids in a lipid carrier may be to protect the nucleic acids from environments that may contain enzymes, chemicals, or conditions that degrade nucleic acids (e.g., RNA). Furthermore, incorporating nucleic acids in a lipid carrier may facilitate nucleic acid uptake and release from endosomal compartments. Therefore, when administered to cells or subjects, it may enhance the therapeutic effects of nucleic acids (e.g., RNA).

[0317] In embodiments, the lipid-based carrier is selected from liposomes, lipid nanoparticles, lipoplexes, solid lipid nanoparticles, lipopolyplexes, and / or nanoliposomes. In preferred embodiments, the lipid-based carrier is lipid nanoparticles (LNPs).

[0318] In a particularly preferred embodiment, the lipid nanoparticles encapsulate at least one nucleic acid, preferably at least one RNA of the present invention.

[0319] In a preferred embodiment, the lipid-based carrier comprises at least one lipid selected from at least one aggregation-inhibiting lipid, at least one cationic lipid, at least one neutral lipid or phospholipid, or at least one steroid or steroid analog, or any combination thereof.

[0320] In a preferred embodiment, the lipid-based carrier comprises (i) an aggregation-inhibiting lipid, (ii) a cationic or ionizable lipid, and (iii) a neutral lipid / phospholipid or a steroid / steroid analog.

[0321] In a particularly preferred embodiment, the lipid-based carrier comprises (i) an aggregation-inhibiting lipid, (ii) a cationic or ionizable lipid, (iii) a neutral or phospholipid, (iv) a steroid or steroid analog.

[0322] Aggregation-inhibiting lipids / polymer-binding lipids: In a preferred embodiment, the lipid-based carrier comprises at least one aggregation-inhibiting lipid or moiety.

[0323] The term "aggregation-inhibiting moisture" refers to a molecule containing moisture suitable for reducing or preventing aggregation of lipid-based carriers. The term "aggregation-inhibiting lipid" refers to a molecule containing both a lipid portion and moisture suitable for reducing or preventing aggregation of lipid-based carriers. Under storage conditions or during formulation, lipid-based carriers may undergo charge-induced aggregation, which can be an undesirable state for the stability of the lipid-based carrier. Therefore, it may be desirable to have a compound or moisture that can inhibit aggregation, for example, by sterically stabilizing the lipid-based carrier. Such steric stabilization may occur when a compound having a sterically bulky, uncharged moisture shields or screens the charged portion of the lipid-based carrier in the composition, thereby preventing the lipid-based carrier from coming into contact with other lipid-based carriers. In the context of the present invention, stabilization of the lipid-based carrier is achieved by including a lipid which may have a sterically bulky group. The lipid is preferably located on the outside of the lipid-based carrier after its formation. Groups suitable for inhibiting aggregation include hydrophilic groups (such as monosialoganglioside GM1), polyamide oligomers (PAO), or specific polymers (such as poly(oxyalkylene), e.g., poly(ethylene glycol) or poly(propylene glycol)).

[0324] Lipids containing polymers as aggregation inhibitory groups are referred to as "polymer-bound lipids" in this specification.

[0325] The term "polymer-bound lipid" refers to a molecule containing both a lipid and a polymer portion, where the polymer is suitable for reducing or preventing aggregation of lipid-based carriers containing RNA. A polymer should be understood as a very large molecule composed of many repeating units, or a substance or material consisting of a macromolecule. In the context of this invention, a suitable polymer may be a hydrophilic polymer. Examples of polymer-bound lipids include PEGylated or PEG-bound lipids.

[0326] In a preferred embodiment, the lipid-based carrier includes an aggregation-inhibiting lipid selected from polymer-bound lipids.

[0327] In preferred embodiments, the polymer-bound lipid is a PEG-bound lipid (or PEGylated lipid, PEG lipid).

[0328] The average molecular weight of PEG moiety in the PEG-bound lipid is preferably in the range of about 500 to about 8,000 daltons (e.g., about 1,000 to about 4,000 daltons). In one preferred embodiment, the average molecular weight of PEG moiety is about 2,000 daltons. The PEG-bound lipid may be selected from or derived from 1,2-dimiristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG2000 DMG or DMG-PEG2000), C10-PEG2K, or Cer8-PEG2K. Alternatively, the PEG-bound lipid may be selected from or derived from formula (IVa) of WO2018078053. A suitable PEG-bound lipid of formula (IVa) has the chemical formula 2[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide (also known as ALC-0159).

[0329] In a preferred embodiment, the lipid-based carrier includes an aggregation-inhibiting lipid, which is not a PEG-bound lipid. Therefore, the aggregation-inhibiting lipid may be appropriately selected from PEG-free lipids (such as PEG-free polymer-bound lipids).

[0330] In a preferred embodiment, the aggregation-inhibiting lipid (or polymer-bound lipid) is a PEG-free lipid containing a polymer different from PEG.

[0331] In the context of the present invention, PEG-free lipids may be selected from or derived from POZ lipids. In preferred embodiments, the POZ lipids or preferred polymer-bound lipids are described in WO2023031394, and the entire disclosure is incorporated by reference. In particular, the disclosures relating to polymer-bound lipids described in any one of claims 1 to 8 of WO2023031394 are incorporated by reference.

[0332] In this embodiment, the polymer-bound lipid is a PEG-free lipid selected from POZ lipids.

[0333] Therefore, in the embodiment, the polymer-bound lipid is a "POZ lipid," preferably defined as a compound of the formula (POZ):[H]-[linker]-[M], In formula (POZ), [H] is a homopolymer molecule containing at least one polyoxazoline (POZ) monomer unit,

[0334] [ka]

[0335] Here, R is a C1-9 alkyl or C2-9 alkenyl, preferably C1, and the average value of n is in the range of 2-200, preferably 20-100, more preferably 24-26 or 45-50; [linker] is any linker group; and [M] is a lipid moisture.

[0336] In preferred embodiments in the context of POZ lipids, the aggregation-inhibiting lipid is selected from or derived from PMOZ1, PMOZ2, PMOZ3, PMOZ4, or PMOZ5 as described in WO2023031394.

[0337] In a preferred embodiment, the aggregation-inhibiting lipid is given by the following formula:

[0338] [ka]

[0339] Selected from PMOZ4 according to, or derived therefrom, In a preferred embodiment, at least one aggregation-inhibiting lipid is selected from DMG-PEG2000, C10-PEG2K, Cer8-PEG2K, or POZ lipids (such as PMOZ4). In another preferred embodiment, at least one aggregation-inhibiting lipid is selected from ALC-0159.

[0340] Cationic lipids: In a preferred embodiment, the lipid-based carrier comprises at least one cationic or ionizable lipid.

[0341] The cationic or ionizable lipids of the lipid-based carrier may be either cationizable or ionizable. That is, when the pH falls below the pK of the lipid's ionizable group, it becomes protonated, while at higher pH values ​​it gradually becomes neutral. At pH values ​​below the pK, the lipid can bind to negatively charged nucleic acids. In certain embodiments, the cationic lipids include zwitterionic lipids that acquire a positive charge as the pH decreases.

[0342] In a preferred embodiment, the lipid-based carrier comprises a cationic or ionizable lipid, which preferably carries a positive charge at physiological pH. More preferably, the cationic or ionizable lipid comprises a tertiary or quaternary nitrogen group. Therefore, in a preferred embodiment, the lipid-based carrier comprises a cationic or ionizable lipid selected from amino lipids, which preferably comprises a tertiary amine group.

[0343] In a preferred embodiment, at least one cationic or ionizable lipid is a lipid of formula (III-1)

[0344] [ka]

[0345] Selected from or derived from Preferably, in formula (III-1), Either 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 L1 or L2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S S-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, -NRaC(=O)NRa-, -OC(=O)NRa-, or -NRaC(=O)O-, or a direct bond; G1 and G2 are independently unsubstituted C1-C12 alkylene or C1-C12 alkenylene; G3 is C1-C24 alkylene, C1-C24 alkenylene, C3-C8 cycloalkylene n is a C3-C8 cycloalkenylene; Ra is H or a C1-C12 alkyl; R1 and R2 are independently a C6-C24 alkyl or a C6-C24 alkenyl; R3 is H, OR5, CN, C(=O)OR4, OC(=O)R4, or -NR5C(=O)R4, where R4 is a C1-C12 alkyl; R5 is H or a C1-C6 alkyl; and x is 0, 1, or 2.

[0346] In some embodiments, cationic or ionizable lipids may be selected from the lipids disclosed in WO2018078053 (i.e., lipids derived from formulas I, II, and III of WO2018078053, or lipids defined in claims 1 to 12 of WO2018078053). The disclosures of WO2018078053 are incorporated in their entirety by reference. In this context, the lipids disclosed in Table 7 of WO2018078053 (e.g., lipids derived from formulas I-1 to I-41) and the lipids disclosed in Table 8 of WO2018078053 (e.g., lipids derived from formulas II-1 to II-36) may be used as appropriate in the context of the present invention. Accordingly, formulas I-1 to I-41 and II-1 to II-36 of WO2018078053, and certain disclosures related thereto, are incorporated herein by reference.

[0347] In embodiments, the lipid-based carrier of the pharmaceutical composition comprises cationic lipids selected from or derived from structures III-1 to III-36 in Table 9 of WO2018078053. Accordingly, formulas III-1 to III-36 of WO2018078053, and certain related disclosures, are incorporated herein by reference.

[0348] In 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 formula ((4-hydroxybutyl)azandiyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) and is also known as ALC-0315 (CAS number 2036272-55-4).

[0349] Furthermore, suitable cationic lipids may be selected from or derived from the cationic lipids listed in PCT claims 1 to 14 of WO2021123332 or in Table 1 of WO2021123332. The disclosures relating to claims 1 to 14 or Table 1 in WO2021123332 are incorporated herein by reference. Accordingly, suitable cationic lipids may be selected from or derived from the cationic lipids corresponding to compounds 1 to 27 (C1 to C27) listed in Table 1 of WO2021123332.

[0350] In some embodiments, the lipid-based carrier is selected from (COATSOME® SS-EC)SS-33 / 4PE-15 (see C23 in Table 1 of WO2021123332) or comprises a cationic lipid derived therefrom. In other embodiments, the lipid-based carrier is selected from HEXA-C5DE-PipSS (see C2 in Table 1 of WO2021123332) or comprises a cationic lipid derived therefrom. In preferred embodiments, the lipid-based carrier is selected from compound C26 disclosed in Table 1 of WO2021123332 or comprises a cationic lipid derived therefrom.

[0351] In other embodiments, the lipid carrier comprises 9-heptadecanyl 8-{(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]aminooctanoic acid (also known as SM-102) or a cationic lipid derived therefrom.

[0352] Therefore, in a preferred embodiment, the lipid-based carrier (preferably LNP) comprises a cationic lipid selected from or derived from the above-mentioned compounds C26, SM-102, SS-33 / 4PE-15, or HEXA-C5DE-PipSS. In another preferred embodiment, the lipid-based carrier (preferably LNP) comprises a cationic lipid selected from or derived from ALC-0315.

[0353] Neutral lipids: In a preferred embodiment, the lipid-based carrier (e.g., LNP) comprises at least one neutral lipid or phospholipid.

[0354] The term "neutral lipid" refers to one of numerous lipid species that exist in an uncharged or neutral zwitterionic form 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-difitanoyl-sn-glycero-3-phosphoethanolamine (DPhyPE), DPhyPS (1,2-difitanoyl-sn-glycero-3-phospho-L-serine), or mixtures thereof.

[0355] In a preferred embodiment, the lipid-based carrier comprises a neutral lipid selected from or derived from DSPC, DHPC, DPhyPE, or DPhyPS. In this context, DSPC is preferred.

[0356] Steroids, steroid analogs, or sterols: In preferred embodiments, the lipid-based carrier comprises a steroid, a steroid analog, or a sterol.

[0357] In embodiments, the steroid, steroid analogue, or sterol is selected from or derived from cholesterol, cholesteryl hemysuccinate (CHEMS), or derivatives thereof. In preferred embodiments, the lipid carrier comprises cholesterol.

[0358] Lipid-based carrier composition: In the embodiment, a lipid-based carrier (preferably LNP) containing at least one nucleic acid (preferably at least one RNA) is, (i) at least one cationic or ionizable lipid, preferably as defined herein; (ii) at least one or two (e.g., two different types) neutral lipids or phospholipids, preferably as defined herein; (iii) at least one steroid or steroid analog, preferably as defined herein; and, (iv) at least one aggregation-inhibiting lipid, preferably as defined herein, Includes.

[0359] In the embodiment, a lipid-based carrier (preferably LNP) containing at least one nucleic acid (preferably at least one RNA) is, (i) At least one cationic lipid selected from or derived from C26, SS-33 / 4PE-15, HEXA-C5DE-PipSS, SM-102; (ii) at least one or two (e.g., two different) neutral lipids selected from or derived from DSPC, DHPC, DPhyPE, and DPhyPS; (iii) at least one steroid or steroid analog selected from or derived from cholesterol; and, (iv) At least one aggregation-inhibiting lipid selected from or derived from DMG-PEG2000, C10-PEG2K, Cer8-PEG2K, ALC-0159, or "PMOZ4"; This includes the lipid carrier, which encapsulates nucleic acids (e.g., RNA).

[0360] In the embodiment, a lipid-based carrier (preferably LNP) containing at least one nucleic acid (preferably at least one RNA) is, (i) at least one cationic lipid selected from or derived from ALC-0315; (ii) at least one neutral lipid selected from or derived from DSPC; (iii) at least one steroid or steroid analog selected from or derived from cholesterol; and, (iv) At least one aggregation-inhibiting lipid selected from or derived from ALC-0159; This includes the lipid carrier, which encapsulates nucleic acids (e.g., RNA).

[0361] In the embodiment, a lipid-based carrier comprising at least one nucleic acid (preferably at least one RNA) is (i) Cationic lipids selected from C26; (ii) Neutral lipids selected from DPhyPE and neutral lipids selected from DPhyPS; (iii) Steroids or steroid analogues selected from cholesterol; and, (iv) Aggregation-inhibiting lipids selected from POZ lipids (preferably PMOZ4) as defined herein; This includes, and here, the lipid-based carrier encapsulates nucleic acids (e.g., RNA).

[0362] In embodiments, cationic lipids, neutral lipids, steroids or steroid analogs, and / or aggregation-inhibiting lipids (all as defined herein) may be combined in various relative proportions.

[0363] In embodiments, the lipid carrier (preferably an LNP) comprises (i) to (iv) in the following molar ratios: about 20 to 60% cationic or ionizable lipids, about 5 to 25% neutral lipids, about 25 to 55% steroids or steroid analogs, and about 0.5 to 15% aggregation-inhibiting lipids (e.g., polymer-binding lipids). Preferably, the lipid carrier encapsulates at least one nucleic acid (preferably at least one RNA).

[0364] In a preferred embodiment, the lipid carrier (preferably an LNP) comprises (i) to (iv) in the following molar ratios: about 45 to 55% cationic or ionizable lipids, about 5 to 15% neutral lipids, about 35 to 45% steroids or steroid analogs, and about 0.5 to 2.5% aggregation-inhibiting lipids (e.g., polymer-bound lipids). Preferably, the lipid carrier encapsulates at least one nucleic acid (preferably at least one RNA).

[0365] In a more preferred embodiment, the lipid carrier (preferably an LNP) comprises (i) to (iv) in the following molar ratios: about 47 to 51% cationic or ionizable lipids, about 8 to 12% neutral lipids, about 38 to 42% steroids or steroid analogs, and about 0.75 to 1.75% aggregation-inhibiting lipids (e.g., polymer-bound lipids). Preferably, the lipid carrier encapsulates at least one nucleic acid (preferably at least one RNA).

[0366] In the embodiment, a lipid-based carrier (preferably LNP) containing at least one nucleic acid (preferably at least one RNA) is, (i) At least one cationic lipid selected from ALC-0315; (ii) Neutral lipids DPhyPE and DPhyPS; (iii) Steroid or steroid-like cholesterol; and, (iv) At least one aggregation-inhibiting lipid selected from ALC-0159; The lipid carrier contains nucleic acids (e.g., RNA). Preferably, (i) to (iv) consist of about 47.4% cationic lipids, about 10% neutral lipids, about 40.9% steroids or steroid analogs, and about 1.7% aggregation-inhibiting lipids, and preferably, the lipid carrier contains nucleic acids (e.g., RNA).

[0367] In a preferred embodiment, a lipid-based carrier (preferably LNP) comprising at least one nucleic acid molecule (preferably RNA) is: (i) Cationic lipid C26 (VitE-C4DE-Pip-thioether); (ii) at least one neutral lipid selected from DSPC; (iii) at least one steroid or steroid analog selected from cholesterol; and, (iv) aggregation-inhibiting lipid PMOZ4; This includes, preferably, (i) to (iv) about 49% cationic lipids, about 10% neutral lipids, about 40% steroids or steroid analogs, and about 1% aggregation-inhibiting lipids, where preferably, the lipid carrier encapsulates at least one nucleic acid (preferably RNA).

[0368] The amount of lipids contained in the lipid carrier may be selected considering the amount of nucleic acid to be loaded. In one embodiment, these amounts are selected such that the N / P ratio of the lipid carrier encapsulating the nucleic acid is in the range of 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 groups of the lipid to the phosphate groups ("P") of the loaded nucleic acid. The N / P ratio may be calculated, for example, based on the fact that 1 μg of nucleic acid typically contains about 3 nmol of phosphate residues, provided that the nucleic acid exhibits a statistical distribution of bases. The "N" value of the lipid or lipidoid may be calculated based on its molecular weight, as well as the relative content of permanent cationic groups and (if present) cationizable groups.

[0369] In some embodiments, the N / P ratio may be in the range of about 1 to about 50. In preferred embodiments, the N / P ratio range is 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.

[0370] In various embodiments, the pharmaceutical composition comprises a lipid-based carrier having a defined size (particle diameter, homogeneous size distribution) (encompassing nucleic acids, preferably RNA as defined herein).

[0371] In this specification, the size of lipid-based carriers is usually described as the Z-mean size. The term "Z-mean size" refers to the average diameter of particles measured by dynamic light scattering (DLS), and the so-called cumulant algorithm is used for data analysis. This analysis yields the so-called Z-mean, which has the dimension of length, and the dimensionless polydispersity index (PDI). The term "dynamic light scattering" or "DLS" refers to a method for analyzing particles in a liquid. In this method, the liquid is usually irradiated with a monochromatic light source, and the light scattered by the particles in the liquid is detected. DLS protocols and apparatus are known in the art.

[0372] In a preferred embodiment, the lipid-based support (preferably LNP) has a Z-average size of less than 400 nm, preferably less than 300 nm, and more preferably less than 200 nm.

[0373] In the embodiment, the lipid-based carrier (preferably LNP) has a Z-average size in the range of about 50 nm to about 200 nm, preferably in the range of about 50 nm to about 150 nm, and more preferably in the range of about 50 nm to about 120 nm.

[0374] Preferably, the composition contains about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and less than 1% of lipid-based carriers with a particle size greater than about 500 nm. Preferably, the composition contains about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and less than 1% of LNPs with a particle size less than about 20 nm.

[0375] In the embodiment, the lipid-based carrier exhibits a zeta potential in the range of -50mV to +50mV, preferably -25mV to +25mV, more preferably -10mV to +10mV, and most preferably -5mV to +5mV.

[0376] In embodiments, the polydispersity index (PDI) of the lipid-based carrier (preferably LNP) is in the range of 0.1 to 0.5. In preferred embodiments, the PDI value is less than about 0.3, preferably less than about 0.2. Typically, the PDI is measured by dynamic light scattering.

[0377] In embodiments, at least 70%, 80%, 90%, or 95% of the nucleic acid (e.g., RNA) is encapsulated in a lipid-based carrier such as an LNP. The encapsulation rate may be measured by a RiboGreen assay known in the art.

[0378] In the embodiment, the lipid-based carriers have a lamellar structure and / or a bilayer structure. In the embodiment, at least 80%, 85%, 90%, and 95% of the lipid-based carriers have a spherical structure.

[0379] In a preferred embodiment, the surface of the lipid-based carrier (preferably LNP) is not charged at pH 7.

[0380] [Administration of pharmaceutical compositions or nucleic acids] Ideally, when the pharmaceutical composition or nucleic acid is administered to cells, tissues, or a target, at least one tumor antigen is produced in an amount sufficient to induce an antigen-specific immune response in those cells, tissues, or the target.

[0381] In preferred embodiments, administration is intramuscular, intratumoral, or intravenous, preferably intramuscular injection, intratumoral injection, or intravenous injection.

[0382] In a preferred embodiment, an encoded tumor antigen is produced when the pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a target. Preferably, the tumor antigen is produced in an amount sufficient to induce an antigen-specific immune response in the cells, tissues, or the target.

[0383] In a preferred embodiment, when a pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a target, the encoded tumor antigen is produced, thereby enhancing immunogenicity in the target.

[0384] In a preferred embodiment, when a pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a target, an encoded tumor antigen is produced, thereby inducing epitope-specific or antigen-specific CD8+ T cells in the target.

[0385] In a preferred embodiment, when a pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a target, an encoded tumor antigen is produced, thereby inducing epitope-specific or antigen-specific CD4+ T cells in the target.

[0386] In a preferred embodiment, when a pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a subject, the encoded tumor antigen is produced, thereby inducing humoral immunity against the tumor antigen in the subject (such as an increase in antibody titer and / or increased diversity of antibody species).

[0387] In a preferred embodiment, when a pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a subject, the encoded tumor antigen is produced, which increases IFN-γ production by CD8+ T cells exposed to the encoded tumor antigen within the subject.

[0388] In a preferred embodiment, when a pharmaceutical composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a target, the encoded tumor antigen is produced, thereby increasing the presentation of the encoded tumor antigen on MHC molecules within the target.

[0389] The "presentation" of encoded polypeptides or peptides refers to an increase in the quantity and / or diversity of immunogenic and / or stable peptides presented via MHC class I and class II molecules after proteolytic degradation by the proteasome mechanism. Therefore, it is associated with cellular immunity (e.g., T cell activation). Presentation of polypeptides or peptides encoded on MHC class I and class II molecules is increased in cells including immune cells (such as T cells), antigen-presenting cells (such as dendritic cells, macrophages, and designed antigen-presenting cells), MHC class I-expressing cells, MHC class II-expressing cells, or any combination thereof.

[0390] For example, administration of a pharmaceutical composition or nucleic acid increases the immunogenic presentation of the encoded tumor antigen on the MHC molecule by approximately 0.1% to approximately 100% (e.g., approximately 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 approximately 100%) compared to subjects not administered the nucleic acid / pharmaceutical composition.

[0391] In some embodiments, administration of a pharmaceutical composition or nucleic acid results in an immunogenicity or presentation of an encoded tumor antigen on MHC molecules that is approximately 2 to 100 times greater (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 greater) than in subjects that do not receive the nucleic acid / pharmaceutical composition.

[0392] 3: Tumor antigen / composition of tumor antigen: In a third embodiment, the present invention provides a peptide tumor antigen, or a composition comprising at least one peptide tumor antigen.

[0393] In particular, the features and embodiments described in the context of the first embodiment (the nucleic acid of the present invention) or the second embodiment (the pharmaceutical composition of the present invention) should be read and understood as appropriate embodiments of the tumor antigen or composition of this embodiment.

[0394] 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 the tumor neoantigen.

[0395] In preferred embodiments in this context, at least one antigenic peptide encoded by a long non-coding RNA (lncRNA) (in particular 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 an immunogenic fragment or variant thereof.

[0396] Preferred peptide tumor antigens are shown in Tables 1A and 1B (column B or C).

[0397] In a preferred embodiment, at least one antigenic peptide is selected from peptides or proteins encoded by long non-coding RNA (lncRNA), At least one amino acid sequence that is identical to any one of sequence numbers 124-146, 546-571, 756, or 757, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, It includes or consists of.

[0398] In a preferred embodiment, at least one antigenic peptide is selected from a tumor neoantigen peptide or protein, in particular 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 an immunogenic fragment or variant thereof.

[0399] Preferred peptide tumor antigens are shown in Table 2 (column C).

[0400] In this embodiment, at least one antigenic peptide selected from the tumor neoantigen peptide or protein is At least one amino acid sequence that is identical to any one of sequence numbers 255-317, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0401] In a preferred embodiment, at least one antigenic peptide selected from the tumor neoantigen peptide or protein is: At least one amino acid sequence that is identical to any one of sequence numbers 255-286, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, It includes or consists of.

[0402] Furthermore, this specification provides compositions comprising at least one peptide tumor antigen described herein.

[0403] "A composition comprising at least one tumor antigen" means any type of composition that may incorporate a specific tumor antigen and may optionally incorporate further components, typically at least one pharmaceutically acceptable carrier or excipient. The composition may be a dry composition (such as a powder, granules, or solid freeze-dried form). Alternatively, the composition may be in liquid form, and each component may be incorporated independently in a dissolved or dispersed form (e.g., suspension or emulsion form).

[0404] In various embodiments, at least one tumor antigen of the composition is formulated with a pharmaceutically acceptable carrier or excipient.

[0405] In embodiments, the composition comprises a plurality 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, preferably 2 to 5 different peptide tumor antigens (all as defined herein).

[0406] 4: Antibodies, T cells, or TCRs: In a fourth aspect, the present invention provides antibodies, T cells, or T cell receptors (TCRs) prepared in response to or stimulated by a tumor antigen or nucleic acid described in any of the first to third aspects.

[0407] In particular, the features and embodiments described in the context of the first embodiment (nucleic acid of the present invention), the second embodiment (pharmaceutical composition of the present invention), or the third embodiment (tumor antigen) should be read and understood as appropriate embodiments of the tumor antigen or nucleic acid of this embodiment.

[0408] In the embodiments, the antibody is a therapeutic antibody produced against any of the tumor antigens of the present invention. Preferably, the antibody is produced against any of the peptide tumor antigens shown in Tables 1A and 1B (column B or C), or any of the peptide tumor antigens shown in Table 2 (column C).

[0409] In the embodiment, the T cells are therapeutic T cells stimulated by the tumor antigen or nucleic acid described in any one of the first to third embodiments. Preferably, the T cells are stimulated by the tumor antigen or nucleic acid shown in Table 1A or 1B or Table 2.

[0410] In the embodiments, T cells were generated in response to the tumor antigen of the present invention (e.g., an antigenic peptide loaded onto MHC). Preferably, the T cells were generated in response to either the peptide tumor antigen shown in Table 1A or B (column B or C), or the peptide tumor antigen shown in Table 2 (column C).

[0411] 5: Combinations: In a fifth embodiment, the present invention provides a combination comprising at least two or more therapeutic modalities described in any one of the first to fourth embodiments.

[0412] In particular, 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, or TCRs) should be read and understood as appropriate embodiments of the combination of the fifth aspect.

[0413] In a preferred embodiment, the combination is: a) at least one artificial nucleic acid as defined in the first embodiment; and / or b) at least one pharmaceutical composition as defined in the second embodiment; and / or, c) at least one tumor antigen or composition as defined in the third embodiment; and / or d) At least one antibody, T cell, or TCR as defined in the fourth aspect; It includes at least two treatment modalities selected from the following.

[0414] In embodiments, the combination of the present invention comprises at least one or more therapeutic modalities as defined herein, preferably 2, 3, 4, 5, 6, 7, 8, or more therapeutic modalities.

[0415] Therapeutic modalities may be administered spatially separated and / or temporally staggered.

[0416] 6: Kit or parts kit: In a sixth embodiment, the present invention provides a kit or a parts kit.

[0417] In particular, 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, or TCRs), or the fifth aspect (combinations) should be read and understood as appropriate embodiments of the kit or component kit of the sixth aspect.

[0418] In a preferred embodiment, the kit or parts kit is i) at least one artificial nucleic acid as defined in the first embodiment; and / or ii) At least one pharmaceutical composition as defined in the second embodiment; and / or, iii) at least one tumor antigen or composition as defined in the third embodiment; and / or iv) At least one antibody, T cell, or TCR as defined in the fourth aspect; Includes.

[0419] In the embodiment, the kit or parts kit includes a liquid vehicle for dissolution.

[0420] In embodiments, the kit or component kit includes technical instructions providing information regarding the administration and dosage of the components. The technical instructions of the kit may include information regarding the administration and dosage of the components, as well as information about the patient group. Such a kit (preferably a component kit) may be applied to any of the applications or uses described herein, for example, and may be applied to the use of therapeutic agents i) to iv) for the treatment of cancer or cancer-related diseases, disorders, or conditions.

[0421] 7: Medical use: In a further embodiment, the present invention relates to the medical use of a therapeutic agent according to any one of the first to sixth embodiments.

[0422] In particular, the features and embodiments described in the context of the kit or component kit described in the first embodiment (nucleic acid of the present invention), the second embodiment (pharmaceutical composition of the present invention), the third embodiment (tumor antigen), the fourth embodiment (antibody, T cell, or TCR), the fifth embodiment (combination), or the sixth embodiment should be read and understood as appropriate embodiments of the medical use shown in the following embodiments.

[0423] Accordingly, the present invention provides, for use as a pharmaceutical (for example, to treat or prevent a target disease, disorder, or condition), an artificial nucleic acid in a first aspect, a pharmaceutical composition in a second aspect, a tumor antigen or composition of a tumor antigen in a third aspect, an antibody, T cell, or TCR in a fourth aspect, a combination in a fifth aspect, or a kit or component kit in a sixth aspect.

[0424] In a preferred embodiment, the use may be for human medical purposes or veterinary medical purposes, and is preferably for human medical purposes.

[0425] In preferred embodiments, the use may be for human medical purposes, particularly for infants, neonates, immunocompromised recipients, pregnant and lactating women, and the elderly.

[0426] In a preferred embodiment, the use may be for human medical purposes in cancer patients.

[0427] [Medical use in the treatment or prevention of cancer] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid; in a second aspect, a pharmaceutical composition; in a third aspect, a tumor antigen or composition of a tumor antigen; in a fourth aspect, an antibody, T cell or TCR; in a fifth aspect, a combination; or in a sixth aspect, a kit or component kit, for use as a pharmacopoeia for treating or preventing cancer or any cancer-related disease, disorder, or condition in a subject.

[0428] In particular, the features and embodiments described in the context of any one of the aforementioned embodiments are This should be interpreted and understood as an appropriate embodiment of this design.

[0429] As used herein, the term “cancer” refers to a neoplasm characterized by uncontrolled and usually rapid cell growth with a tendency to invade surrounding tissues and metastasize to distant sites. This term encompasses both benign and malignant neoplasms. Malignancy in cancer is typically characterized by dysplasia, invasiveness, and metastasis. Benign neoplasms, on the other hand, typically do not possess these characteristics. In addition to neoplasms characterized by tumor growth, this term includes cancers of the blood and lymphatic systems. Furthermore, the term “cancer” in this disclosure also includes cancer metastases (e.g., HNSCC, NSCLC, or cancer metastases associated with melanoma).

[0430] In some embodiments, the treatment or prevention of cancer in a subject is personalized therapy. Therefore, the patient's cancer is analyzed, for example, by sequencing, before treatment with each therapeutic agent.

[0431] In various embodiments, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations thereof, kits, or component kits are administered intramuscularly, intratumorally, or intravenously, particularly by injection.

[0432] [Medical use in the treatment or prevention of squamous cell carcinoma] In a further embodiment, the present invention provides, for use as a pharmaceutical for treating or preventing squamous cell carcinoma, an artificial nucleic acid in a first embodiment, a pharmaceutical composition in a second embodiment, a tumor antigen or composition of a tumor antigen in a third embodiment, an antibody, T cell or TCR in a fourth embodiment, a combination in a fifth embodiment, or a kit or component kit in a sixth embodiment.

[0433] In particular, the features and embodiments described in the context of any one of the aforementioned embodiments are This should be interpreted and understood as an appropriate embodiment of this design.

[0434] In a preferred embodiment, squamous cell carcinoma is selected from pulmonary squamous cell carcinoma (LUSC) / squamous non-small cell lung cancer (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC).

[0435] In some embodiments, the treatment or prevention of squamous cell carcinoma in a subject is personalized therapy. Therefore, the squamous cell carcinoma of the patient is analyzed, for example, by sequencing, before treatment with each therapeutic agent.

[0436] [Medical use in the treatment or prevention of NSCLC] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit for use as a pharmacopoeia to treat or prevent non-small cell lung cancer (NSCLC) or any disease, disorder or condition associated with NSCLC in a subject.

[0437] In embodiments, this application relates to the treatment of NSCLC subgroups (including lung adenocarcinoma (LUAD) and / or lung squamous cell carcinoma (LUSC)). Preferably, this application relates to the treatment of lung squamous cell carcinoma (LUSC) / sqNSCLC.

[0438] In particular, the features and embodiments described in the context of any one of the aforementioned embodiments are This should be interpreted and understood as an appropriate embodiment of this design.

[0439] In preferred embodiments in this context, an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or at least one tumor antigen provided by a kit or component kit is: a) Peptides or proteins 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 those immunogenic fragments or any of those variants, At least one antigenic peptide selected from; and / or, b) A peptide or protein of a tumor neoantigen selected from BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, PIK3CA neoantigen, or NFE2L2 neoantigen, Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Preferably, at least one antigenic peptide selected from EGFR neoantigen or TP53 neoantigen, or an immunogenic fragment or variant thereof. Includes.

[0440] In a particularly preferred embodiment in this context, an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or at least one tumor antigen provided by a kit or component kit is: Peptides or proteins 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 those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes.

[0441] In various embodiments of this context, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations thereof, kits, or component kits are administered by intramuscular, intratumoral, or intravenous administration, particularly by intramuscular injection.

[0442] In some embodiments, the treatment or prevention of NSCLC in a subject is individualized therapy. Therefore, the patient's NSCLC cancer is analyzed, for example, by sequencing, before treatment with each therapeutic agent.

[0443] [Medical use in the treatment or prevention of HNSCC] In a further embodiment, the present invention provides, in a first embodiment, an artificial nucleic acid; in a second embodiment, a pharmaceutical composition; in a third embodiment, a tumor antigen or composition of a tumor antigen; in a fourth embodiment, an antibody, T cell or TCR; in a fifth embodiment, a combination; or in a sixth embodiment, a kit or component kit, for use as a pharmacopoeia to treat or prevent head and neck squamous cell carcinoma (HNSCC) or any disease, disorder or condition associated with HNSCC in a subject.

[0444] In particular, the features and embodiments described in the context of any one of the aforementioned embodiments are This should be interpreted and understood as an appropriate embodiment of this design.

[0445] In a particularly preferred embodiment in this context, an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or at least one tumor antigen provided by a kit or component kit is: Peptides or proteins encoded by lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes.

[0446] In various embodiments of this context, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations thereof, kits, or component kits are administered by intramuscular, intratumoral, or intravenous administration, particularly by intramuscular injection.

[0447] In some embodiments, the treatment or prevention of HNSCC in a subject is individualized therapy. Therefore, the patient's HNSCC cancer is analyzed, for example, by sequencing, before treatment with each therapeutic agent.

[0448] [Medical use in the treatment or prevention of melanoma] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid; in a second aspect, a pharmaceutical composition; in a third aspect, a tumor antigen or composition of a tumor antigen; in a fourth aspect, an antibody, T cell, or TCR; in a fifth aspect, a combination; or in a sixth aspect, a kit or component kit, for use as a pharmacopoeia for treating or preventing melanoma or any disease, disorder, or condition associated with melanoma in a subject.

[0449] In embodiments, this application relates to the treatment of cutaneous melanoma (SKCM) or uveal melanoma.

[0450] In this embodiment, the application relates to the treatment of the BRAF subgroup and / or NRAS subgroup of melanoma.

[0451] In particular, the features and embodiments described in the context of any one of the aforementioned embodiments are This should be interpreted and understood as an appropriate embodiment of this design.

[0452] In preferred embodiments in this context, an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or at least one tumor antigen provided by a kit or component kit is: a) A peptide or protein encoded by lnc-TRPC5-3:1 or lnc-CLEC2D-9:1, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from; and / or, b) A peptide or protein of a tumor neoantigen 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, NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, or BRAF neoantigen, Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes.

[0453] In a particularly preferred embodiment in this context, an artificial nucleic acid, a pharmaceutical composition, a tumor antigen or a composition of tumor antigens, or at least one tumor antigen provided by a kit or component kit is: A peptide or protein encoded by lnc-TRPC5-3:1 or lnc-CLEC2D-9:1, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes.

[0454] In various embodiments of this context, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations thereof, kits, or component kits are administered by intramuscular, intratumoral, or intravenous administration, particularly by intramuscular injection.

[0455] In some embodiments, the treatment or prevention of melanoma in a subject is individualized therapy. Therefore, the patient's melanoma is analyzed, for example, by sequencing, before treatment with each therapeutic agent.

[0456] [Medical use in the treatment or prevention of further applicable cancer types] In a further embodiment, the present invention relates to a medical use of the therapeutic agent described in any one of the above embodiments for treating or preventing further indications for cancer.

[0457] In particular, the features and embodiments described in the context of the kit or component kit described in the first embodiment (nucleic acid of the present invention), the second embodiment (pharmaceutical composition of the present invention), the third embodiment (tumor antigen), the fourth embodiment (antibody, T cell, or TCR), the fifth embodiment (combination), or the sixth embodiment should be read and understood as appropriate embodiments of the medical use shown in the following embodiments.

[0458] In a further aspect, the present invention provides, in a first aspect, an artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell, or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit, for use as a pharmaceutical to treat or prevent adrenocortical carcinoma, urothelial carcinoma of the bladder, invasive breast cancer, cervical cancer (cervical squamous cell carcinoma and cervical adenocarcinoma), colon adenocarcinoma, renal papillary cell carcinoma, lung adenocarcinoma, lymphoid neoplasm diffuse large B cell lymphoma, rectal adenocarcinoma, sarcoma (especially synovial sarcoma), cutaneous melanoma, thymoma, thyroid cancer, hepatocellular carcinoma, uterine carcinosarcoma, or endometrial cancer of the uterine body, or any disease, disorder, or condition related thereto. Herein, the artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell, or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit. A peptide or protein encoded by ZC3H8-6:1, or The immunogenic fragment or its variant, It contains at least one antigenic peptide selected from the following.

[0459] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell, or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit, for use as a pharmaceutical to treat or prevent bladder urothelial carcinoma, cervical cancer (cervical squamous cell carcinoma and cervical adenocarcinoma), thymoma, or endometrial cancer of the uterine body, or any disease, disorder, or condition related thereto. Herein, at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of a tumor antigen, the combination, or the kit or component kit is A peptide or protein encoded by KCNMB2-AS1:4, or The immunogenic fragment or its variant, It contains at least one antigenic peptide selected from the following.

[0460] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell, or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit for use as a pharmacopoeia to treat or prevent ovarian serous cystic carcinoma, testicular germ cell tumor, thymoma, or uterine carcinosarcoma, or any disease, disorder, or condition related thereto. Herein, at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of a tumor antigen, the combination, or the kit or component kit is A peptide or protein encoded by lnc-NTF3-5:5, or The immunogenic fragment or its variant, It contains at least one antigenic peptide selected from the following.

[0461] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell, or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit for use as a pharmacopoeia for treating or preventing bladder urothelial carcinoma, colon adenocarcinoma, esophageal carcinoma, or gastric adenocarcinoma, or any disease, disorder, or condition related thereto in a subject. Herein, at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of a tumor antigen, the combination, or the kit or component kit is A peptide or protein encoded by lnc-WDR72-2:4, or The immunogenic fragment or its variant, It contains at least one antigenic peptide selected from the following.

[0462] In a further embodiment, the present invention provides, in a first aspect, an artificial nucleic acid, in a second aspect, a pharmaceutical composition, in a third aspect, a tumor antigen or composition of a tumor antigen, in a fourth aspect, an antibody, T cell, or TCR, in a fifth aspect, a combination, or in a sixth aspect, a kit or component kit for use as a pharmacopoeia for treating or preventing cutaneous melanoma, testicular germ cell tumor, or uveal melanoma, or any disease, disorder, or condition related thereto in a subject. Herein, at least one tumor antigen provided by the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of a tumor antigen, the combination, or the kit or component kit is A peptide or protein encoded by lnc-CLEC2D-9:1, or The immunogenic fragment or its variant, It contains at least one antigenic peptide selected from the following.

[0463] 8: Treatment method: In a further embodiment, the present invention relates to a method for treating or preventing a disease, disorder, or condition.

[0464] In particular, embodiments relating to the aforementioned aspects should be read and understood as appropriate embodiments of the therapeutic method of the present invention. In particular, certain features and embodiments of the therapeutic method described herein may also be applied to the medical use of the present invention, and vice versa.

[0465] Disease prevention (suppression) or treatment concerns suppressing the complete onset of a disease or condition in individuals at risk of disease (such as infectious diseases or cancer). “Treatment” refers to therapeutic interventions that improve the signs or symptoms of a disease or condition after it has begun to develop. The term “improvement” in reference to a disease or condition refers to an observable and beneficial effect of treatment. Disease suppression may include preventing or reducing the risk of the disease. Such beneficial effects may be demonstrated, for example, by delaying the onset of clinical symptoms of the disease in susceptible individuals, reducing the severity of some or all clinical symptoms of the disease, slowing the rate of disease progression, improving the overall health or well-being of the individual, or by other parameters specific to the particular disease. “Preventive” treatment is treatment given to individuals who show no signs of disease or only early signs, with the aim of reducing the risk of the condition developing.

[0466] In a preferred embodiment, the present invention relates to a method for treating or preventing a disease, disorder, or condition. The method includes applying or administering an effective amount of an artificial nucleic acid according to the first embodiment, a pharmaceutical composition according to the second embodiment, a tumor antigen or composition of a tumor antigen according to the third embodiment, an antibody, T cells, or TCR according to the fourth embodiment, a combination according to the fifth embodiment, or a kit or component kit according to the sixth embodiment to a subject in need thereof.

[0467] In this specification, when referring to the amount of a therapeutic compound, “effective” means an amount of the compound sufficient to produce the desired therapeutic effect without excessive adverse side effects (such as toxicity, irritation, or allergic reactions) that are commensurate with a reasonable benefit-to-risk ratio, when used in the manner of this disclosure.

[0468] In a preferred embodiment, the disease, disorder, or condition is cancer (e.g., any disease, disorder, or condition related to cancer as defined herein). In a preferred embodiment, the cancer is NSCLC. In a preferred embodiment, the cancer is squamous cell carcinoma, preferably selected from pulmonary squamous cell carcinoma (LUSC) / squamous non-small cell lung cancer (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC). In another preferred embodiment, the cancer is melanoma.

[0469] Preferred embodiments in this context are, in particular, described in the sections “Medical use in the treatment or prevention of squamous cell carcinoma,” “Medical use in the treatment or prevention of NSCLC,” “Medical use in the treatment or prevention of HNSCC,” “Medical use in the treatment or prevention of melanoma,” or “Medical use in the treatment or prevention of further applicable cancer species,” as outlined in the context of “Medical use.”

[0470] In various embodiments, nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations thereof, kits, or component kits are administered intramuscularly, intratumorally, or intravenously, particularly by injection.

[0471] In some embodiments, the treatment or prevention of cancer in a subject is personalized therapy. Therefore, the patient's cancer is analyzed, for example, by sequencing, before treatment with each therapeutic agent.

[0472] Item list Preferred embodiments of the present invention are shown in the following numbered list (items 1 to 116).

[0473] 1. An artificial nucleic acid comprising at least one coding sequence encoding the at least one tumor antigen, wherein the tumor antigen is a) A peptide or protein encoded by a long non-coding RNA (lncRNA), or at least one antigenic peptide selected from an immunogenic fragment or variant thereof; and / or b) A peptide or protein of a tumor neoantigen, or at least one antigenic peptide selected from its immunogenic fragment or variant thereof. Artificial nucleic acids containing or consisting of.

[0474] 2. At least one antigenic peptide, Peptides or proteins encoded by long non-coding RNAs (lncRNAs) 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 those immunogenic fragments or any of those variants, Artificial nucleic acids as described in item 1, selected from the options provided.

[0475] 3. The artificial nucleic acid according to items 1-2, wherein the at least one antigenic peptide encoded by the long non-coding RNA (lncRNA) comprises at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0476] 4. The at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA) is At least one amino acid sequence that is identical to any one of sequence numbers 124-254, 546-650, or 756-763, or at least one that is identical to any one of these by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those immunogenic fragments or any of those variants, Artificial nucleic acids as described in items 1-3, including or consisting of the following.

[0477] 5. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by lnc-WDR72-2:4, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 124-146, 546-571, 756, or 757, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0478] 6. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from a peptide or protein encoded by lnc-TRPC5-3:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 227-237, 630-642, or 761, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0479] 7. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-CLEC2D-9:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 238-243, 643-650, or 762, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0480] 8. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by KCNMB2-AS1:4, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 147-161, 572-589, or 758, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0481] 9. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-ZC3H8-6:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 162-168, 590-603, or 759, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0482] 10. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-NTF3-5:5, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 169-226, 604-629, or 760, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0483] 11. At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by LINC00893:25, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 244-254 or 763, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of Artificial nucleic acids as described in items 1-4, including those listed above.

[0484] 12. The at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from a peptide or protein encoded by an lncRNA, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 318-448, 651-755, or 764-771, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or It is a fragment of any of those or a variant of any of those. Artificial nucleic acids as described in items 1-11.

[0485] 13. The at least one coding sequence comprises a GC-optimized nucleic acid sequence encoding at least one antigenic peptide selected from a peptide or protein encoded by an lncRNA, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 318-448, 651-755, or 764-771, or it is at least 80% identical to any one of these, or It is a fragment of one of those, Artificial nucleic acids as described in item 12.

[0486] 14. The artificial nucleic acid described in item 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, CNOT9 neoantigen, or immunogenic fragments thereof or variants thereof.

[0487] 15. The artificial nucleic acid described in item 14, wherein at least one antigenic peptide selected from the tumor neoantigen peptide or protein comprises at least one T cell epitope, preferably at least one CD8+ T cell epitope.

[0488] 16. At least one antigenic peptide selected from the tumor neoantigen peptide or protein, At least one amino acid sequence that is identical to any one of sequence numbers 255-317, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, Artificial nucleic acids as described in item 14 or 15, including or consisting of the following.

[0489] 17. comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of NRAS tumor neoantigens, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 267-272, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0490] 18. comprising at least one coding sequence encoding at least one antigenic peptide selected from the peptide or protein of the TYW1B tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 273 or 274, or at least 80% identical to either of these, Those immunogenic fragments or their variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0491] 19. comprising at least one coding sequence encoding at least one antigenic peptide selected from ECPAS tumor neoantigen peptides or proteins, Here, the antigenic peptide is At least one amino acid sequence that is identical to, or at least 80% identical to, SEQ ID NO: 275, or The immunogenic fragment or its variant, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0492] 20. Consists of at least one coding sequence encoding at least one antigenic peptide selected from the peptide or protein of the MAP2K1 tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NO: 276 or 277, or at least 80% identical to either of these, Those immunogenic fragments or their variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0493] 21. comprising at least one coding sequence encoding at least one antigenic peptide selected from the TOMM22 tumor neoantigen peptide or protein, Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 278 or 279, or at least 80% identical to either of these, Those immunogenic fragments or their variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0494] 22. comprising at least one coding sequence encoding at least one antigenic peptide selected from the peptide or protein of the GLB1 tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NO: 280 or 281, or at least 80% identical to either of these, Those immunogenic fragments or their variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0495] 23. Consists of at least one coding sequence encoding at least one antigenic peptide selected from the peptides or proteins of the MAGE-A3 tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to either SEQ ID NOs. 282 or 283, or at least 80% identical to either of these, Those immunogenic fragments or their variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0496] 24. comprising at least one coding sequence encoding at least one antigenic peptide selected from the peptide or protein of the ATAD2 tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 284-286, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0497] 25. comprising at least one coding sequence encoding at least one antigenic peptide selected from peptides or proteins of BRAF tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 262-266, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0498] 26. comprising at least one coding sequence encoding at least one antigenic peptide selected from the peptides or proteins of EGFR tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 255-257, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0499] 27. TP53 comprises at least one coding sequence encoding at least one antigenic peptide selected from the peptide or protein of the tumor neoantigen, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 258-261, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, Artificial nucleic acids as described in items 14-16, including or consisting of the following.

[0500] 28. The at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from a tumor neoantigen peptide or protein, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 449-511, or is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or It is a fragment of any of those or a variant of any of those. Artificial nucleic acids as described in items 14-27.

[0501] 29. The at least one coding sequence comprises a GC-optimized nucleic acid sequence encoding at least one antigenic peptide selected from the peptide or protein of a tumor neoantigen, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 449-511, or is at least 80% identical to any one of these, or An artificial nucleic acid, as described in item 28, which is a fragment of one of those.

[0502] 30. The artificial nucleic acid according to any one of items 1 to 29, wherein at least one coding sequence codes for 2 to 10 different tumor antigens, preferably 2 to 5 different tumor antigens.

[0503] 31. Artificial nucleic acids as described in item 30, in which different tumor antigens are separated by linker elements.

[0504] 32. An artificial nucleic acid according to any one of items 1 to 31, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one immune response activation signaling protein.

[0505] 33. An artificial nucleic acid according to any one of items 1 to 32, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one T helper epitope.

[0506] 34. An artificial nucleic acid according to any one of items 1 to 33, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one signal peptide.

[0507] 35. The artificial nucleic acid according to any one of items 1 to 34, wherein the at least one coding sequence is a codon-modified coding sequence, preferably, where the codon-modified coding sequence is selected from a C-maximizing coding sequence, a CAI-maximizing coding sequence, a coding sequence adapted for human codon use, a G / C content-modified coding sequence, and a G / C-optimized coding sequence, or any combination thereof.

[0508] 36. An artificial nucleic acid as described in item 35, wherein at least one codon-modified coding sequence is a G / C optimized coding sequence.

[0509] 37. The artificial nucleic acid according to any one of items 1 to 36, 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.

[0510] 38. The at least one heterogeneous 3'-UTR is 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 selected from any one homolog, fragment, or variant of those genes, including or consisting of Preferably, here, the heterogeneous 3'-UTR is Nucleic acid sequences that are identical to sequence numbers 66-95, 112-123, or are identical to them by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, Artificial nucleic acids as described in item 37, including or consisting of the following.

[0511] 39. The at least one heterologous 3'-UTR comprises or consists of a nucleic acid sequence selected from or derived from the 3'-UTR of the PSMB3 gene, wherein the heterologous 3'-UTR is Nucleic acid sequences identical to sequence numbers 66, 67, 112-123, or identical to them by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, preferably the nucleic acid sequence of sequence number 67, or Any of those fragments or any of those variants, Artificial nucleic acids as described in item 37 or 38, including or consisting of the following.

[0512] 40. The at least one heterogeneous 5'-UTR is 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 selected from any one homolog, fragment, or variant of those genes, including or consisting of Preferably, here, the heterogeneous 5'-UTR is Nucleic acid sequences that are identical to sequence numbers 12-45, 64, and 65, or are identical to them by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, Artificial nucleic acids as described in item 37, including or consisting of the following.

[0513] 41. The at least one heterologous 5'-UTR comprises or consists of a nucleic acid sequence selected from or derived from the 5'-UTR of the HSD17B4 gene, wherein the at least one heterologous 5'-UTR is Nucleic acid sequences identical to sequence numbers 12, 13, 64, and 65, or identical to them by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, preferably the nucleic acid sequence of sequence number 13, or Any of those fragments or any of those variants, Artificial nucleic acids as described in item 37 or 40, including or consisting of the following.

[0514] 42. The artificial nucleic acid according to any one of items 37 to 41, wherein at least one heterogeneous 5'-UTR is selected from HSD17B4 and at least one heterogeneous 3'-UTR is selected from PSMB3.

[0515] 43. The artificial nucleic acid according to any one of items 1 to 42, wherein the artificial nucleic acid is selected from DNA or RNA, preferably from RNA.

[0516] 44. The artificial nucleic acid described in any one of items 1 to 43, wherein the artificial nucleic acid is RNA selected from mRNA, circular RNA, replicon RNA, or viral RNA.

[0517] 45. The artificial nucleic acid described in any one of items 1 to 44, wherein the artificial nucleic acid is mRNA.

[0518] 46. ​​The artificial nucleic acid according to any one of items 1 to 45, wherein the artificial nucleic acid (preferably the RNA) comprises at least one poly(A) sequence, preferably, the at least one poly(A) sequence comprises about 40 to about 500 adenosine nucleotides.

[0519] 47. The artificial nucleic acid according to item 46, wherein the at least one poly(A) sequence comprises about 60 to about 150 adenosine nucleotides, preferably about 100 adenosine nucleotides.

[0520] 48. The artificial nucleic acid described in item 46 or 47, wherein at least one poly(A) sequence is located at the 3' end, where optionally the nucleotide at the 3' end is adenosine.

[0521] 49. The artificial nucleic acid according to any one of items 1 to 48, 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.

[0522] 50. The artificial nucleic acid (preferably the RNA) comprises at least one histone stem-loop sequence, where the histone stem-loop sequence is A nucleic acid sequence that is identical to either sequence number 3 or 4, or is at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to either of these sequences, or Those fragments or their variants, Artificial nucleic acids as described in any one of items 1 to 49, including or consisting of.

[0523] 51. The artificial nucleic acid according to any one of items 1 to 50, wherein the artificial nucleic acid is an RNA comprising at least one modified nucleotide, preferably comprising a modified nucleotide selected from pseudouridine (ψ) or N1-methylpseudridine (m1ψ).

[0524] 52. The artificial nucleic acid described in any one of items 1 to 51, wherein the artificial nucleic acid is a modified RNA, where each uracil is replaced by a modified nucleotide.

[0525] 53. The artificial nucleic acid described in item 51 or 52, wherein the modified nucleotide is N1-methylpseuduridine (m1ψ).

[0526] 54. The artificial nucleic acid described in items 1 to 50, wherein the nucleic acid is RNA that does not contain modified nucleotides.

[0527] 55. The artificial nucleic acid described in any one of items 1 to 54, wherein the artificial nucleic acid is RNA containing a 5'-cap structure.

[0528] 56. The artificial nucleic acid according to item 55, wherein the 5'-cap structure is selected from a cap-1 structure or a modified cap-1 structure.

[0529] 57. An artificial nucleic acid as described in any one of items 1 to 56, wherein the artificial nucleic acid is an in vitro transcribed RNA.

[0530] 58. The artificial nucleic acid according to any one of items 1 to 57, wherein the artificial nucleic acid is purified RNA, preferably by at least one step of RP-HPLC, AEX, SEC, hydroxyapatite chromatography, TFF, filtration, precipitation, core bead flow-through chromatography, oligo(dT) purification, cellulosic purification, or any combination thereof.

[0531] 59. The artificial nucleic acid according to any one of items 1 to 58, wherein the artificial nucleic acid (preferably the RNA) has an integrity of at least 50%, preferably 60%, more preferably 70%, and most preferably 80%.

[0532] 60. The artificial nucleic acid (preferably RNA) according to any one of items 1 to 59, which is suitable for use in the treatment or prevention of a disease, disorder, or condition (preferably a tumor disease, disorder, or condition).

[0533] 61. The artificial nucleic acid (preferably the RNA) preferably has the following sequence elements in the 5' to 3' direction: A) 5'-cap structure; B) Preferably a 5'-UTR selected from the 5'-UTR of the HSD17B4 gene; C) A coding sequence encoding at least one tumor antigen; D) Preferably a 3'-UTR selected from the PSMB3 gene; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides, Artificial nucleic acids as described in any one of items 1 through 60, including those listed in item 1 through 60.

[0534] 62. The artificial nucleic acid is preferably mRNA containing the following sequence elements in the 5' to 3' direction: A) 5'-cap structure; B) A nucleic acid sequence that is identical to sequence number 13, or at least 80% identical to it, The fragment or variant thereof, A 5'-UTR that includes or consists of; C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA), wherein the nucleic acid sequence is identical to, or at least 80% identical to, any one of sequence numbers 318-448, 651-755, or 764-771, or is a fragment or variant of any of them; D) A nucleic acid sequence that is identical to sequence number 67, or at least 80% identical to it, The fragment or variant thereof, 3'-UTR containing or consisting of; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides, Artificial nucleic acids as described in items 1-61.

[0535] 63. The artificial nucleic acid is preferably mRNA containing the following sequence elements in the 5' to 3' direction: A) 5'-cap structure; B) A nucleic acid sequence that is identical to sequence number 13, or at least 80% identical to it, The fragment or variant thereof, A 5'-UTR that includes or consists of; C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from the peptides or proteins of tumor neoantigens, wherein the nucleic acid sequence is identical to any one of sequence numbers 449-511, or at least 80% identical to any one of these, or is a fragment or variant of any of these; D) A nucleic acid sequence that is identical to sequence number 67, or at least 80% identical to it, The fragment or variant thereof, 3'-UTR containing or consisting of; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides, Artificial nucleic acids as described in items 1-61.

[0536] 64. A pharmaceutical composition comprising at least one artificial nucleic acid, The artificial nucleic acid is an artificial nucleic acid as defined in any one of items 1 to 63, comprising at least one coding sequence encoding at least one tumor antigen. Pharmaceutical composition.

[0537] 65. The pharmaceutical composition according to item 64, comprising a plurality of artificial nucleic acids, each encoding at least one different tumor antigen.

[0538] 66. The pharmaceutical composition according to item 64 or 65, wherein the at least one artificial nucleic acid (preferably the RNA) is formulated within at least one cationic or polycationic compound.

[0539] 67. The pharmaceutical composition according to item 66, wherein the at least one cationic or polycationic compound is selected from a cationic or polycationic polymer, a cationic or polycationic polysaccharide, a cationic or polycationic lipid, a cationic or polycationic protein, a cationic or polycationic peptide, or any combination thereof.

[0540] 68. The pharmaceutical composition according to items 64-67, wherein the at least one artificial nucleic acid (preferably the RNA) is formulated within a lipid carrier.

[0541] 69. The pharmaceutical composition according to item 68, wherein the lipid-based carrier is selected from liposomes, lipid nanoparticles, lipoplexes, solid lipid nanoparticles, lipopolyplexes, and / or nanoliposomes.

[0542] 70. The pharmaceutical composition according to item 68 or 69, wherein the lipid carrier is lipid nanoparticles.

[0543] 71. The pharmaceutical composition according to items 68-70, wherein the lipid carrier comprises at least one aggregation-inhibiting lipid, at least one cationic or ionic lipid, at least one neutral or phospholipid, and at least one steroid or steroid analog.

[0544] 72. The pharmaceutical composition according to item 71, wherein the aggregation-inhibiting lipid is a polymer-bound lipid selected from PEG-bound lipids or PEG-free lipids.

[0545] 73. The pharmaceutical composition according to item 72, wherein the polymer-bound lipid is selected from DMG-PEG2000, C10-PEG2K, Cer8-PEG2K, or POZ lipids.

[0546] 74A. The pharmaceutical composition according to items 71 to 73, wherein the cationic lipid or ionic lipid is selected from amino lipids, and preferably the amino lipid contains a tertiary nitrogen group.

[0547] 74B. Cationic lipids or ionic lipids are selected from or derived from formula (III-1, as defined herein), preferably either L1 or L2 being -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 L1 or L2 being -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S S-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, -NRaC(=O)NRa-, -OC(=O)NRa-, or -NRaC(=O)O-, or a direct bond; G1 and G2 are independently unsubstituted C1-C12 alkylene or C1-C12 alkenylene; G3 is C1-C24 alkylene, C1-C24 alkenylene, C3-C8 cycloalkylene R1 is C3-C8 cycloalkenylene; Ra is H or C1-C12 alkyl; R1 and R2 are independently C6-C24 alkyl or C6-C24 alkenyl; R3 is H, OR5, CN, C(=O)OR4, OC(=O)R4, or -NR5C(=O)R4, R4 is C1-C12 alkyl; R5 is H or C1-C6 alkyl; and x is 0, 1, or 2. Pharmaceutical compositions as described in items 71-74A.

[0548] 74C. The cationic lipid is selected from or derived from formula III-3 of WO2018078053, preferably having the chemical formula ((4-hydroxybutyl)azandiyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) (also known as ALC-0315).

[0549] 75. A pharmaceutical composition according to items 71-74A, 74B, or 74C, wherein the cationic lipid or ionic lipid is selected from ALC-0315, SM-102, SS-33 / 4PE-15, HEXA-C5DE-PipSS, or compound C26.

[0550] 76. The pharmaceutical composition according to items 71 to 75, wherein the neutral lipid or phospholipid is selected from DSPC, DHPC, or DPhyPE.

[0551] 77. The pharmaceutical composition according to items 71-76, wherein the steroid or steroid analog is selected from cholesterol and cholesteryl hemysuccinate (CHEMS), preferably cholesterol.

[0552] 78. The lipid-based carrier (preferably LNP) (i) at least one cationic lipid, preferably as defined in item 74 or 75; (ii) at least one neutral lipid, preferably as defined in item 76; (iii) at least one steroid or steroid analog, preferably one as defined in item 77; and, (iv) at least one aggregation-inhibiting lipid, preferably one as defined in item 72 or 73, Pharmaceutical compositions as described in items 68-77, including the above.

[0553] 79. The pharmaceutical composition according to items 68 to 78, wherein the lipid 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% aggregation-inhibiting lipids.

[0554] 80. The pharmaceutical composition according to items 68 to 79, wherein the weight ratio of lipids to nucleic acids (e.g., RNA) in the lipid-based carrier is approximately 10:1 to approximately 60:1.

[0555] 81. The pharmaceutical composition according to items 68 to 80, wherein the N / P ratio of the lipid carrier for encapsulating nucleic acids (preferably RNA) is in the range of about 1 to about 20.

[0556] 82. The pharmaceutical composition according to items 68 to 81, wherein the lipid-based carrier has a Z-average size in the range of about 50 nm to about 200 nm.

[0557] 83. A pharmaceutical composition according to items 64-82, further comprising at least one RNA sensor-type pattern recognition receptor antagonist, selected from Toll-like receptor antagonists, preferably a TLR7 antagonist and / or a TLR8 antagonist, most preferably the RNA of Sequence ID No. 85 described in WO2021028439.

[0558] 84. A pharmaceutical composition or artificial nucleic acid as described in any one of items 1 to 83, wherein the composition or nucleic acid produces an encoded tumor antigen when administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects.

[0559] 85. A pharmaceutical composition or artificial nucleic acid described in any one of items 1 to 84, wherein when the composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects, an encoded tumor antigen is produced, thereby inducing epitope-specific CD8+ T cells in the subject.

[0560] 86. A tumor antigen or a composition of at least one tumor antigen characterized by any one of items 1 to 85, preferably any one of items 2 to 4 or 14 to 16.

[0561] 87. The tumor antigen is At least one amino acid sequence that is identical to any one of sequence numbers 124-254, 546-650, or 756-763, or at least one that is identical to any one of these by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those immunogenic fragments or any of those immunogenic variants, A tumor antigen or composition as described in item 86, comprising or consisting of the following.

[0562] 88. The tumor antigen is At least one amino acid sequence that is identical to any one of sequence numbers 255-317, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, A tumor antigen or composition as described in item 86, comprising or consisting of the following.

[0563] 89. The tumor antigen or composition according to items 86-88, wherein the composition comprises a plurality of tumor antigens, and optionally, the antigens are formulated in a pharmaceutically acceptable carrier.

[0564] 90. Antibodies, T cells, or TCRs produced against or stimulated by any tumor antigen or nucleic acid as defined in any one of items 1 through 89.

[0565] 91. a) At least one artificial nucleic acid as described in items 1-62; and / or, b) At least one pharmaceutical composition described in items 63-85; and / or, c) At least one tumor antigen or composition as described in items 86-89; and / or, d) At least one antibody, T cell, or TCR as described in item 90. A combination of at least two or more treatment modalities selected from the above.

[0566] 92. A compound comprising at least one artificial nucleic acid as described in any one of items 1 to 62, and / or at least one pharmaceutical composition as described in any one of items 63 to 85, and / or at least one tumor antigen or tumor antigen composition as described in items 86 to 89, and / or at least one antibody, T cell, or TCR as described in item 90. It optionally contains a liquid vehicle for dissolution, and, Optionally includes technical instructions providing information on the administration and dosage of the ingredients. A kit or parts kit.

[0567] 93. Artificial nucleic acids as described in any one of items 1 to 62, pharmaceutical compositions as described in any one of items 63 to 85, tumor antigens or compositions of tumor antigens as described in items 86 to 89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92.

[0568] 94. Artificial nucleic acids as described in any one of items 1 to 62, pharmaceutical compositions as described in any one of items 63 to 85, tumor antigens or compositions of tumor antigens as described in items 86 to 89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as a medicine to treat or prevent cancer or any cancer-related disease, disorder or condition in a subject.

[0569] 95. An artificial nucleic acid as described in any one of items 1 to 62, a pharmaceutical composition as described in any one of items 63 to 85, a tumor antigen or composition of tumor antigen as described in items 86 to 89, an antibody, T cell or TCR as described in item 90, a combination as described in item 91, or a kit or component kit as described in item 92, for use as a pharmaceutical to treat or prevent squamous cell carcinoma or any disease, disorder or condition associated with squamous cell carcinoma in a subject, preferably the squamous cell carcinoma is selected from LUSC / squamous non-small cell lung cancer (sqNSCLC) or head and neck squamous cell carcinoma (HNSCC).

[0570] 96. Artificial nucleic acids as described in any one of items 1 to 62, pharmaceutical compositions as described in any one of items 63 to 85, tumor antigens or compositions of tumor antigens as described in items 86 to 89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as a medicine to treat or prevent NSCLC or any disease, disorder or condition related to NSCLC in a subject.

[0571] 97. Use as a pharmaceutical for the treatment or prevention of NSCLC as described in item 96, wherein the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigens, the combination, or at least one tumor antigen provided by a kit or component kit, (a) Peptides or proteins 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 those immunogenic fragments or any of those variants, At least one antigenic peptide selected from; and / or, (b) Peptides or proteins of BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, PIK3CA neoantigen, NFE2L2 neoantigen, preferably peptides or proteins selected from EGFR neoantigen or TP53 neoantigen, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes, use.

[0572] 96B. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as a medicine to treat or prevent HNSCC or any disease, disorder or condition associated with HNSCC in a subject.

[0573] 97B. Use as a pharmaceutical for treating or preventing HNSCC as described in item 96B, wherein the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigens, the combination, or at least one tumor antigen provided by a kit or component kit, (a) Peptides or proteins encoded by lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes, use.

[0574] 98. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as a medicine to treat or prevent melanoma or any disease, disorder or condition related to melanoma in a subject.

[0575] 99. Use as a pharmaceutical for the treatment or prevention of melanoma as described in item 98, wherein the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigens, the combination, or at least one tumor antigen provided by a kit or component kit, (a) A peptide or protein encoded by lnc-CLEC2D-9:1, lnc-TRPC5-3:1, or LINC00893:25, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from; and / or, (b) Peptides or proteins derived 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, and CNOT9 neoantigen. Preferably, a peptide or protein selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, or BRAF neoantigen, Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, Includes, use.

[0576] 99B. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as pharmaceuticals to treat or prevent adrenocortical carcinoma, urothelial carcinoma of the bladder, invasive breast cancer, cervical cancer (cervical squamous cell carcinoma and cervical adenocarcinoma), colon adenocarcinoma, renal papillary cell carcinoma, lung adenocarcinoma, lymphoid neoplasms, diffuse large B cell lymphoma, rectal adenocarcinoma, sarcoma, cutaneous melanoma, thymoma, thyroid cancer, hepatocellular carcinoma, uterine carcinosarcoma, or endometrial cancer of the uterine body, or any disease, disorder or condition related thereto, in the subject, Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-ZC3H8-6:1, or comprising at least one antigenic peptide selected from its immunogenic fragment or variant thereof, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations, or kits or component kits.

[0577] 99C. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as pharmaceuticals to treat or prevent urothelial carcinoma of the bladder, cervical cancer (squamous cell carcinoma of the cervix and adenocarcinoma of the cervix), thymoma, or endometrial cancer of the uterine body, or any disease related thereto, in the subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by KCNMB2-AS1:4, or comprising at least one antigenic peptide selected from its immunogenic fragment or variant thereof, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations, or kits or component kits.

[0578] 99D. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as pharmaceuticals to treat or prevent ovarian serous cystic carcinoma, testicular germ cell tumor, thymoma, or uterine carcinosarcoma, or any disease related thereto, in the subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-NTF3-5:5, or comprising at least one antigenic peptide selected from its immunogenic fragment or variant thereof, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations, or kits or component kits.

[0579] 99E. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as pharmaceuticals to treat or prevent bladder urothelial carcinoma, colon adenocarcinoma, esophageal carcinoma, or gastric adenocarcinoma, or any disease related thereto, in a subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-WDR72-2:4, or comprising at least one antigenic peptide selected from its immunogenic fragment or variant thereof, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells, or TCRs, combinations, or kits or component kits.

[0580] 99F. Artificial nucleic acids as described in items 1-62, pharmaceutical compositions as described in items 63-85, tumor antigens or compositions of tumor antigens as described in items 86-89, antibodies, T cells or TCRs as described in item 90, combinations as described in item 91, or kits or component kits as described in item 92, for use as a medicine to treat or prevent cutaneous melanoma, testicular germ cell tumor, or uveal melanoma, or any disease, disorder, or condition related thereto in a subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-CLEC2D-9:1, or It contains at least one antigenic peptide selected from the immunogenic fragment.

[0581] 100. The pharmaceutical use described in items 93-99F, in which the pharmaceutical is administered to the subject by intramuscular, intratumoral, or intravenous administration.

[0582] 101. A method for treating or preventing a disease, disorder, or condition, the method comprising applying or administering to a subject in need of such treatment or prevention an effective amount of an artificial nucleic acid as described in items 1 to 62, a pharmaceutical composition as described in items 63 to 85, a tumor antigen or composition of a tumor antigen as described in items 86 to 89, an antibody, T cell, or TCR as described in item 90, a combination as described in item 91, or a kit or component kit as described in item 92.

[0583] 102. The method according to item 101, wherein the disease, disorder, or condition is cancer.

[0584] 103. The method according to item 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).

[0585] 104. The cancer is NSCLC, wherein the nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, or at least one tumor antigen provided by the kit or component kit is (a) Peptides or proteins 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 those immunogenic fragments or any of those variants, At least one antigenic peptide selected from; and / or, (b) Peptides or proteins derived from BRAF neoantigen, EGFR neoantigen, TP53 neoantigen, PIK3CA neoantigen, or NFE2L2 neoantigen, preferably peptides or proteins selected from EGFR neoantigen or TP53 neoantigen, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, The method described in item 102 or 103, including the method described in item 102 or 103.

[0586] 105. The cancer is melanoma, wherein the nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, or at least one tumor antigen provided by a kit or component kit is (a) A peptide or protein encoded by lnc-CLEC2D-9:1, lnc-TRPC5-3:1, or LINC00893:25, or Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from; and / or, (b) Peptides or proteins derived 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, and CNOT9 neoantigen. Preferably, a peptide or protein selected from NRAS neoantigen, TYW1B neoantigen, ECPAS neoantigen, MAP2K1 neoantigen, TOMM22 neoantigen, GLB1 neoantigen, MAGE-A3 neoantigen, ATAD2 neoantigen, or BRAF neoantigen, Any of those immunogenic fragments or any of those variants, At least one antigenic peptide selected from, The method described in item 102, including the method described in item 102.

[0587] 106. The method described in item 101, characterized by any one of the characteristics of a medical use as defined in items 93-100.

[0588] [Brief explanation of the drawing] [Figure 1] Figure 1 shows an exemplary workflow for neoantigen identification. *Immunogenicity prediction: neoIM; MHC presentation prediction: MHCnuggets & neoMS. **Considering host gene expression, sample coverage, HLA coverage / distribution, MHC presentation ability, and immunogenicity, etc.

[0589] [Figure 2] Figure 2 shows the high tumor specificity of the lncRNA transcript lnc-CLEC2D-9:1 (corresponding smORF: lncCLEC2D91412616). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high target expression in melanoma and low or almost undetectable expression in healthy tissues (excluding the testes, which are immune-privileged tissues).

[0590] [Figure 3] Figure 3 shows the high tumor specificity of the lncRNA transcript lnc-TRPC5-3:1 (corresponding smORF: lncTRPC53116031801). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high targeted expression in melanoma and low or almost undetectable expression in healthy tissue.

[0591] [Figure 4] Figure 4 shows the high tumor specificity of the lncRNA transcript lnc-TRPC5-3:1 (corresponding smORF: lncTRPC53116031801). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high target expression in NSCLC (adenocarcinoma of the lung (LUAD) and squamous cell carcinoma of the lung (LUSC)) and low or almost undetectable expression in healthy tissue.

[0592] [Figure 5] Figure 5 shows the high tumor specificity of the lncRNA transcript lnc-WDR72-2:4 (corresponding smORF: lncWDR722415641696 and lncWDR722416421696). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high target expression in NSCLC (squamous cell carcinoma of the lung (LUSC)) and low or almost undetectable expression in healthy tissue.

[0593] [Figure 6] Figure 6 shows the high tumor specificity of the lncRNA transcript KCNMB2-AS1:4 (corresponding smORF: KCNMB2AS14633753). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high target expression in NSCLC (squamous cell carcinoma of the lung (LUSC)) and low or almost undetectable expression in healthy tissue.

[0594] [Figure 7] Figure 7 shows the high tumor specificity of the lncRNA transcript lnc-NTF3-5:5 (corresponding smORF: lncNTF35511951417). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high target expression in NSCLC (squamous cell carcinoma of the lung (LUSC)) and low or almost undetectable expression in healthy tissue.

[0595] [Figure 8] Figure 8 shows the high tumor specificity of the lncRNA transcript lnc-ZC3H8-6:1 (corresponding smORF: lncZC3H86163144). RNA-seq data obtained from the TCGA (tumor tissue) and GTEX (healthy tissue) databases show high target expression in NSCLC (adenocarcinoma of the lung (LUAD) and squamous cell carcinoma of the lung (LUSC)) and low or almost undetectable expression in healthy tissue.

[0596] [Figure 9] Figure 9 shows that the translated peptide of smORF lncTRPC53116031801 is almost absent in healthy tissues. Mass spectrometry data shows extremely low or almost undetectable expression levels (asterisks) in various healthy tissues compared to highly expressed controls (e.g., housekeeping genes; dots).

[0597] [Figure 10] Figure 10 shows the in vitro antigen validation of melanoma neoantigen (peptide pool 1) in a DC-T cell assay (see Example 6.1, Tables E3-E5 for details). IFN-γ measurements of all T cells after two stimulations (R1 and R2) were obtained for 10 PBMC donors (D1-D10). Antigen-specific T cell responses to at least one peptide from pool 1 were confirmed in 7 donors. Recall and memory responses were observed in one donor each.

[0598] [Figure 11] Figure 11 shows the in vitro antigen validation of melanoma neoantigen (peptide pool 2) in a DC-T cell assay (see Example 6.1, Tables E3-E5 for details). IFN-γ measurements of all T cells after two stimulations (R1 and R2) were obtained for 10 PBMC donors (D1-D10). Antigen-specific T cell responses to at least one peptide from pool 2 were confirmed in 7 donors. Memory responses were observed in 2 donors.

[0599] [Figure 12] Figure 12 shows the in vitro antigen validation of melanoma neoantigen (peptide pool 3) in a DC-T cell assay (see Example 6.1, Tables E3-E5 for details). IFN-γ measurements of all T cells after two stimulations (R1 and R2) were obtained for 10 PBMC donors (D1-D10). Antigen-specific T cell responses to at least one peptide from pool 3 were confirmed in all donors. Recall and memory responses were observed in four donors, each.

[0600] [Figure 13] Figure 13 shows the in vitro antigen validation of melanoma neoantigen (peptide pool 4) in a DC-T cell assay (see Example 6.1, Tables E3-E5 for details). IFN-γ measurements of all T cells after two stimulations (R1 and R2) were obtained for 10 PBMC donors (D1-D10). Antigen-specific T cell responses to at least one peptide from pool 4 were confirmed in 8 donors. Recall and memory responses were observed in 2 and 1 donors, respectively.

[0601] [Figure 14] Figure 14 shows the in vitro antigen validation of melanoma neoantigen (peptide pool 5) in a DC-T cell assay (see Example 6.1, Tables E3-E5 for details). IFN-γ measurements of all T cells after two stimulations (R1 and R2) were obtained for 10 PBMC donors (D1-D10). Antigen-specific T cell responses to the peptides in pool 5 were confirmed in 4 donors. Recall and memory responses were observed in 1 and 2 donors, respectively.

[0602] [Figure 15] Figure 15 shows an overview of all recall and memory responses observed in Example 6.1 / Figures 10-14.

[0603] [Figure 16] Figure 16 shows the in vitro antigen validation of the smORFs shown in the DC-T cell assay (see Example 6.2, Tables E6-E8 for details). IFN-γ measurements of all T cells after two stimulations (R1 and R2) were obtained for various PBMC donors.

[0604] [Figure 17] Figure 17 shows the in vitro antigen validation of melanoma neoantigen in the DC-T cell assay (see Example 6.3, Tables E9-E10A for details). IFN-γ levels of all T cells after two stimulations (R1 and R2) were obtained for various PBMC donors.

[0605] [Figure 18] Figure 18 shows the in vivo immunogenicity of the LNP-formulated mRNA vaccine R12300, which encodes two smORFs (some of which are expressed in melanoma), in CB6F1 hybrid mice. Mice were intramuscularly vaccinated with 5 μg of LNP-formulated R12300 on days 0, 7, and 14. Vaccination with an unrelated mRNA (PpLuc; R8730) served as a control group. Splenocytes were isolated from sacrificial mice on day 21. Splenocytes were restimulated with a 15-mer peptide library against lncCLEC2D91412616 or DMSO (control) and analyzed by flow cytometry. The magnitude of the CD8+ (top) and CD4+ (bottom) T cell responses to the encoded smORF peptide lncCLEC2D91412616 is shown as the percentage of IFN-γ+TNF+ cells in CD8+ or CD4+ T cells, respectively. Median values ​​are plotted.

[0606] [Figure 19] Figure 19 shows the in vivo immunogenicity of the LNP-formulated mRNA vaccine R12301, which encodes multiple smORFs (some of which are expressed in NSLSCs), in CB6F1 hybrid mice. Mice were intramuscularly vaccinated with 5 μg of LNP-formulated R12301 on days 0, 7, and 14. Vaccination with an unrelated mRNA (PpLuc; R8730) served as a control group. Splenocytes were isolated by sacrificing mice on day 21. Splenocytes were individually restimulated with peptide libraries for each smORF (15-mer libraries covering all antigens) or DMSO (control) and analyzed by flow cytometry. The magnitudes of CD8+ (top panel) and CD4+ (bottom panel) T cell responses to the four encoded smORF peptides KCNMB2AS14633753, lncNTF35511951417, lncWDR722415641696, and lncZC3H86163144 (NKZW-smORFs) are shown as the percentage of IFN-γ+TNF+ cells in CD8+ or CD4+ T cells, respectively. Median values ​​are plotted.

[0607] [Examples] Examples illustrating various embodiments of the present invention are presented below. However, the scope of the present invention should not be limited by the specific embodiments presented herein, but rather should be understood to be applicable to other compositions or uses as defined herein. Accordingly, the following preparation methods and examples are provided so that those skilled in the art may better understand and implement the present invention. In fact, various modifications of the present invention, in addition to those described herein, will be readily apparent to those skilled in the art from the foregoing description, the accompanying drawings, and the following examples.

[0608] [Example 1: Antigen discovery and selection method] The objective of this embodiment was to identify cancer antigens that produce epitopes in patients with NSCLC or melanoma. The analysis focused on two types of targets: tumor-specific antigens (neoepitopes) derived from somatic mutations, and tumor-associated antigens encoded by small open reading frames (smORFs) identified within long non-coding RNAs (lncRNAs). The identified smORFs / lncRNAs showed clear tumor-associated expression using transcriptomics and proteomics, and were detected only at low levels in normal tissues. Candidates for both types of antigens were selected using major histocompatibility complex (MHC) binding prediction and immunogenicity prediction.

[0609] Discovery of neoepitopes: Tumor-specific variants were detected using sequence alignment map (BAM) files from the Cancer Genome Atlas (TCGA) database. The most promising candidates were selected using a combination of parameters. A threshold of 0.55 was used for the immunogenicity score (neoim_score, higher values ​​indicate higher immunogenicity). Furthermore, only genes exhibiting at least 20 TPM (transcripts per million) expression were considered. Finally, neoantigens were prioritized based on expected population coverage (proportion of patients with the mutation and predicted MHC-I binding allele). The neoantigen identification workflow is shown in Figure 1.

[0610] Discovery of smORF in lncRNA: To identify smORFs within lncRNAs, the coding potential within the lncRNA sequence was predicted as an initial step. Subsequently, differential transcriptome expression analysis was performed to compare tumor expression with normal expression; only transcripts with a log2 (fold change) greater than 2 and an adjusted p-value less than 1e-4 were selected for further analysis. In addition, MHC-I binding and immunogenicity were predicted for nmers 8-11 of the smORFs, and only smORFs with nmers that were predicted to bind to MHC-I and had a neoim_score greater than 0.5 were retained. Furthermore, only lncRNAs with evidence of translation confirmed by mass spectrometry (PXD009630: melanoma, PXD002612: lung cancer) were included. As a final selection step, lncRNAs / smORFs showing low expression in normal tissue were retained using transcriptomics (GTEx V7) and mass spectrometry (PXD010154).

[0611] The immunogenicity score (neoim_score) is generated using a machine learning algorithm and reflects the probability that the epitope exhibits immunogenicity. RNA-seq expression is shown in TPM; TPM normalizes RNA-seq reads by the length and sequencing depth of each gene and is widely used for comparing expression levels in different samples. Predicted coverage of neoantigens was calculated by product of the population frequency of the mutation and the frequency of the HLA allele to which epitope binding is predicted. HLA allele prediction was performed using a machine learning algorithm.

[0612] [Example 2: Antigen discovery and selection of melanoma and NSCLC neoantigens] The objective of this experiment was to discover melanoma and NSCLC cancer antigens derived from tumor-specific neoantigens. The top hits obtained are summarized in Table 3.

[0613] Table 3 shows the identified neoantigens for melanoma and NSCLC. For each neoantigen in the table, the RNA-seq expression level (mean expression), predicted immunogenicity score (neoim_score), coverage in each cancer type (mean predicted coverage), the HLA allele to which binding is expected (HLA mupexi allele), and the applicable cancer type and subcohort are shown (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 more specifically refers to cutaneous melanoma (SKCM).

[0614] [Table 4] JPEG2026518273000008.jpg161169

[0615] [Example 3: Discovery and selection of melanoma and NSCLC antigens encoded by lncRNA] The objective of this experiment was to discover melanoma and NSCLC cancer antigens derived from lncRNA-smORF. The top hits obtained are summarized in Table 4.

[0616] Table 4 shows the identified melanoma and NSCLC antigens encoded by lncRNAs. For each smORF in the table, the RNA-seq expression level (mean expression), predicted immunogenicity score (neoim_score), and applicable cancer species and subcohort are shown (M=melanoma; N=NSCLC; LUSC=squamous cell carcinoma of the lung (sqNSCLC); LUAD=adenocarcinoma of the lung). In this context, M / melanoma more specifically refers to cutaneous melanoma (SKCM).

[0617] [Table 5]

[0618] In addition, data from mass spectrometry analysis (including 50 melanoma patients (PXD009630-melanoma) and 57 lung cancer patients (PXD002612-lung adenocarcinoma)) confirmed that preferred smORFs were actually translated. In this context, Table 4A shows the frequency of patients in whom smORF translation peptides were detected from tumor samples by mass spectrometry.

[0619] [Table 6]

[0620] [Example 4: High tumor specificity of preferred lncRNA] The objective of this experiment was to comparatively analyze the expression of preferred lncRNA transcripts in tumor tissue and healthy tissue (Table E1). For this purpose, RNA-seq expression data from the TCGA (tumor tissue) and GTEX (healthy tissue) databases were analyzed. Furthermore, mass spectrometry data were also analyzed.

[0621] [result] 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 low or almost undetectable expression in healthy tissues. As an exception, lnc-CLEC2D-9:1 expression was observed in the testes, but this was considered negligible as the testes are an immune-privileged site. The results are shown in Figures 2-8 and summarized in Table E1.

[0622] [Table 7]

[0623] Furthermore, analysis by mass spectrometry confirmed that the preferred smORF-translated peptide showed low or almost undetectable expression in healthy tissue. Representative data for lncTRPC53116031801 are shown in Figure 9.

[0624] Overall, these data suggest that preferred lncRNAs and preferredly derived smORFs are highly expressed at high frequencies in tumors, while their expression is low or almost undetectable in healthy tissues.

[0625] [Example 5: Expression of lncRNA transcripts in the thymus] The objective of this experiment was to analyze the expression of preferred lncRNA transcripts (Table E1) in the thymus. Generally, antigen expression in the thymus is associated with central tolerance, and therefore, for target antigens used in vaccines, ideally, expression should be low or barely detectable. For this purpose, transcriptome analysis was performed using publicly available datasets.

[0626] [result] Representative data from the three datasets shown in Table E2 indicate low or almost undetectable expression of lncRNA transcripts in the thymus. This suggests that smORFs derived from these lncRNA transcripts, such as the preferred smORFs listed in Table E1, are unlikely to undergo central tolerance.

[0627] [Table 8]

[0628] [Example 6: Antigen Verification - In vitro Immunogenicity] 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 described in Examples 1-5.

[0629] Peptides containing the target epitope were synthesized according to standard procedures. The amino acid sequences of these peptides are described in each example. Peptide pools (Examples 6.1 and 6.2) or single peptides (Examples 6.3 and 6.4) were used.

[0630] The immunogenicity of the test peptide was evaluated using an in vitro DC-T cell assay. In this assay, cryopreserved PBMCs from multiple (healthy) donors with a series of HLA types were removed from the cryopreservation chamber, and monocytes were isolated. The cultured monocytes were differentiated into immature dendritic cells (iDCs) using a cytokine cocktail (GM-CSF and IL-4). The iDCs were loaded with the test peptide and further differentiated into mature dendritic cells (mDCs) using CD40L. Subsequently, the mDCs were co-cultured for 8 days with pre-isolated autologous whole T cells in the presence of the cytokine cocktail. During co-culture, the culture medium and cytokines were periodically changed. After 8 days, the whole T cells were harvested (Round 1 = R1), re-stimulated with peptide-loaded monocytes, and then co-cultured for another 8 days in the presence of the cytokine cocktail (Round 2 = R2). After Rounds 1 and 2, the whole T cells were harvested and re-stimulated with peptide-loaded monocytes on an IFN-γ FluoroSpot plate. After overnight stimulation, the FluoroSpot plates were developed according to the manufacturer's protocol. The number of total IFN-γ-secreting T cells was measured using a Mabtech IRIS® FluoroSpot reader. Furthermore, quality control of moDCs was performed using flow cytometry. In addition to the target peptide, appropriate negative controls (DMSO or a control pool of 29 peptides derived from myelin oligodendrocyte glycoprotein (MOG)) and positive controls (MART, CEF / CEFBA) were used. The materials had to be sterile, with endotoxin levels less than 0.1 EU / mg and protein concentrations greater than 1 mg / ml.

[0631] For analysis, IFNγ-positive spot-forming units (SFUs) were measured for each R1 and R2 using the Fluorospot assay. After normalizing SFU / well to SFU / million, δSFU (dSFU) was calculated as the absolute difference in the number of spots between the tested smORF-derived peptide and each DMSO control condition.

[0632] In Examples 6.1 and 6.2 (pooled peptides), the following criteria apply: i) The dSFU of R1 is at least 30, and ii) DSFU R2 is at least twice as high as dSFU R1. If the following conditions were met, the response was considered a "recall response".

[0633] In Example 6.3 (single peptide), the following criteria apply: i) The dSFU of R1 is at least 5, and ii) DSFU R2 is at least twice as high as dSFU R1. If the following conditions were met, the response was considered a "recall response".

[0634] The following criteria: i) The dSFU of R1 is at least 100, and, ii) DSFU R2 is 0.1 to 2.0 times higher than dSFU R1. If the following conditions were met, the response was considered a "memory response".

[0635] It should be noted that failure to meet the above criteria under specific conditions does not necessarily mean that no recall or memory responses were present at all. Rather, it may mean that the magnitude of the recall or memory response was below the threshold, or that it did not meet the above criteria selected for consistent batch analysis.

[0636] [Example 6.1: Antigen Verification - In vitro immunogenicity of melanoma cancer antigen - Pooled peptide] The objective of this experiment was to analyze the in vitro immunogenicity of multiple melanoma cancer antigens derived from tumor-specific neoantigens and encoded by lncRNA-derived smORFs.

[0637] In this assay, cryopreserved PBMCs from 10 different (healthy) donors with a range of HLA types (see Table E3) were tested using a pool of five peptides derived from melanoma cancer antigens (see Table E4).

[0638] [Table 9]

[0639] [Table 10]

[0640] [result] The in vitro immunogenicity of melanoma cancer antigens using the peptide pool shown in Table E4 is shown in Figures 10-14. The graphs show the IFN-γ measurements of the assays described in this example. A schematic of the recall and memory responses, defined according to the above criteria for consistent batch analysis, is shown in Figure 15 and Table E5.

[0641] [Table 11]

[0642] As shown in Figure 10, seven PBMC donors showed an antigen-specific T cell response to at least one peptide in Pool 1. One PBMC donor showed a recall response to at least one peptide in Pool 1. One PBMC donor showed a memory response to at least one peptide in Pool 1 (see also Figure 15 / Table E5). As shown in Figure 11, seven PBMC donors showed an antigen-specific T cell response to at least one peptide in Pool 2. Two PBMC donors showed a memory response to at least one peptide in Pool 2 (see also Figure 15 / Table E5). As shown in Figure 12, ten PBMC donors showed an antigen-specific T cell response to at least one peptide in Pool 3. Four PBMC donors showed a recall response to at least one peptide in Pool 3. Four PBMC donors showed a memory response to at least one peptide in Pool 3 (see also Figure 15 / Table E5). As shown in Figure 13, eight PBMC donors showed an antigen-specific T cell response to at least one peptide in Pool 4. Two PBMC donors showed a recall response to at least one peptide in Pool 4. One PBMC donor showed a memory response to at least one peptide in Pool 4 (see also Figure 15 / Table E5). As shown in Figure 14, four PBMC donors showed an antigen-specific T cell response to at least one peptide in Pool 5. One PBMC donor showed a recall response to at least one peptide in Pool 5. Two PBMC donors showed a memory response to at least one peptide in Pool 5 (see also Figure 15 / Table E5).

[0643] In summary, each peptide pool contains at least one cancer antigen capable of eliciting an immune response in human T cells. Both recall and memory responses were observed in this experimental setup.

[0644] [Example 6.2: Antigen validation - In vitro immunogenicity of NSCLC and melanoma cancer antigens - Pooled peptides] The objective of this experiment was to analyze the in vitro immunogenicity of multiple NSCLC and melanoma cancer antigens derived from tumor-specific neoantigens and encoded by lncRNA-derived smORFs.

[0645] In this assay, cryopreserved PBMCs derived from 10 different (healthy) donors with a range of HLA types (see Table E6), and eight peptide pools derived from NSCLC and melanoma cancer antigens (see Table E7) were tested.

[0646] [Table 12]

[0647] [Table 13]

[0648] [result] Representative in vitro immunogenicity results for the NSCLC / melanoma cancer antigen and peptide pool shown in Table E7 are presented in Figure 15. The graph shows the IFN-γ measurements for the assays described in this example. The recall and memory responses shown in Figure 15 are further summarized in Table E8.

[0649] [Table 14]

[0650] In the condition pool 8 / donor D9 of Example 6.2, an immune response characterized by a dSFU R1 of 21.25 and a dSFU R2 of 236 was observed.

[0651] In summary, the peptides encoded by lncRNAs (lncCLEC2D91412616, KCNMB2AS14633753, lncZC3H86163144, lncNTF35511951417, and lncWDR722415641696) were immunogenic. In particular, the in vitro immunogenicity of the lncRNA-encoded peptides was observed to be equivalent to or greater than that of the well-known immunogenic TAA MART-1 (lncNTF35511951417 and lncWDR722415641696), further highlighting their potential as cancer targets.

[0652] [Example 6.3: Antigen Verification - In vitro immunogenicity of melanoma cancer antigen - Single peptide] The objective of this experiment was to analyze the in vitro immunogenicity of multiple melanoma cancer antigens derived from tumor-specific neoantigens and encoded by lncRNAs.

[0653] We tested cryopreserved PBMCs derived from five (healthy) donors with a range of HLA types (see Table E9), as well as nine single peptides derived from melanoma cancer antigen (see Table E10).

[0654] [Table 15]

[0655] [Table 16]

[0656] In future experiments, we will test additional single peptides from pools 1 and 2 in Table E7.

[0657] [result] Figure 17 shows representative in vitro immunogenicity results for melanoma cancer antigens using single peptides, as shown in Table E10. The graph shows the IFN-γ measurements for the assay described in this example. The recall responses shown in Figure 17 are further summarized in Table E10A.

[0658] [Table 17]

[0659] [Example 6.4: Antigen Verification - In vitro immunogenicity of NSCLC and melanoma cancer antigens - Single peptide] The objective of this experiment was to analyze the in vitro immunogenicity of multiple NSCLC cancer antigens derived from tumor-specific neoantigens and encoded by lncRNAs. In this assay, cryopreserved PBMCs from multiple (healthy) donors with a range of HLA types were tested using a single NSCLC peptide selected from Table E7.

[0660] [Example 7: Antigen Verification - Immunopeptidomics] To further investigate and confirm epitope presentation on MHC-I / II in tumor cells, immunopeptidogenic analysis will be performed. For this purpose, non-targeted mass spectrometry will be performed using primary samples from 15 melanoma patients (including those with BRAF mutations) and 15 NSCLC patients (10 LUSC + 5 LUAD) according to standard procedures in this art. In addition, whole-genome sequencing, RNA-seq, and Ribo-seq will be performed on each sample.

[0661] [Example 8: Preparation of nucleic acids encoding cancer antigens] This embodiment provides a method for obtaining RNA according to the present invention, and a method for producing a composition according to the present invention (containing nucleic acids (particularly RNA) formulated on a lipid-based carrier).

[0662] [Example 8.1: Preparation of DNA template for in vitro RNA transcription] DNA sequences encoding the cancer antigens of the present invention were prepared and used in subsequent in vitro RNA transcription reactions. Some DNA sequences were prepared by modifying wild-type or reference coding DNA sequences by introducing G / C optimized coding sequences for stabilization and expression optimization. The sequences were introduced into pUC-derived DNA vectors and included a stabilized UTR sequence, a stretch of adenosines, and optional histone stem-loops (hSLs). The resulting plasmid DNA templates were transformed and amplified in bacteria using common protocols known in the art. Finally, the plasmid DNA templates were extracted, purified, and linearized using type II restriction enzymes. The RNA constructs used herein are shown in Table E11.

[0663] [Table 18]

[0664] [Table 19]

[0665] [Example 8.2: In vitro transcription of RNA from plasmid DNA template] DNA-dependent RNA in vitro transcription (IVT) was performed using a linearized DNA template with T7 RNA polymerase. During this process, a sequence-optimized nucleotide mixture (ATP / GTP / CTP / UTP) and a cap analog (for Cap1: m7G(5')ppp(5')(2'OMeA)pG; TriLink) were present under appropriate buffering conditions. Other constructs were produced in the presence of nucleic acid mixtures ((ATP / GTP / CTP / pseuduridine(ψ)) or (ATP / GTP / CTP / N1-methylpseuduridine(m1ψ))) and cap analogs. After RNA in vitro transcription, the resulting RNA IVT reactions were subjected to purification steps including RP-HPLC.

[0666] [Example 8.3: Preparation of lipid-based carrier containing mRNA] The LNPs used in this embodiment were prepared 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 via a custom-designed microfluidic mixing chip that allows for the mixing of nanoparticle components on a nanoliter scale in milliseconds. For the preparation of the lipid nanoparticle composition, the following excipients / lipids were used: (i) ionizable lipids: VitE-C4DE-Pip-thioether (compound C26) as described herein and in Table 1 of published patent application WO2021123332; (ii) cholesterol as described herein (Avanti Polar Lipids; Alabaster, AL); (iii) neutral lipids / phospholipids "DPhyPE" (1,2-difitanoyl-sn-glycero-3-phosphoethanolamine; Avanti Polar Lipids; Alabaster, AL) as described herein; (iv) phosphatidylserine DPhyPS (1,2-difitanoyl-sn-glycero-3-phospho-L-serine; Avanti Polar Lipids; Alabaster, AL, 850408P) as described herein; and (v) polymer-bound lipids, PMOZ-lipids "PMOZ4" as described herein. The LNP composition was further characterized according to the method described in Table E13.

[0667] [Table 20]

[0668] Lipids were dissolved in an alcoholic solution (ethanol) according to a standard procedure. Specifically, LNPs were prepared by mixing appropriate amounts of lipid stock solutions (cholesterol, phospholipids, and polymer-bound lipids) in ethanol buffer (20 mg / ml in ethanol). Cationic lipid C26 was dissolved in 30 mg / ml t-butanol and added to the lipid ethanol premixture. Subsequently, this lipid ethanol mixture was mixed with an aqueous phase (50 mM sodium acetate, pH 4.0) containing mRNA (1 g / L). Briefly, the mRNA shown in this example was diluted to 0.05-0.2 mg / ml in 50 mM acetate buffer (pH 4.0). A syringe pump was placed at the inlet of a NanoAssemblr™ (Precision NanoSystems Inc., Vancouver, BC) and used to mix the ethanol lipid solution and the mRNA aqueous solution in a ratio of approximately 1:5-1:3 (volume ratio). The total flow rate was approximately 14 ml / min to 18 ml / min. Afterward, ethanol was removed, and the external buffer was replaced with PBS / sucrose buffer (pH 7.4, 75 mM NaCl, 10 mM phosphate, 150 mM sucrose) by dialysis (Slide-A-Lyzer® dialysis cassette, ThermoFisher). Finally, the lipid nanoparticles were filtered through a sterile filter with a pore size of 0.2 μm. The particle size of the lipid nanoparticles was approximately 90 nm to 140 nm, as measured by quasi-elastic light scattering using a Malvern Zetasizer Nano (Malvern Instruments Ltd.; UK).

[0669] [Example 9: Translation of mRNA encoding smORF and stability of the translated protein] The translation and expression protein stability of mRNA encoding smORF (mRNA constructs listed in Table E11) were measured using Western blotting and flow cytometry according to standard procedures known in the art. HEK293T cells (400,000 cells / well, 200,000 cells / ml) were transfected with mRNA (1 μg / ml) using lipofectamin. After incubation for 4 hours and 16-18 hours with and without carfilzomib, the amount of expressed protein was measured by FACS using antibodies against the CTLA4 domain or HA tag. In addition, HeLa cells were transfected with mRNA. Two hours after transfection, the cells were divided into groups supplemented with or without carfilzomib and treated accordingly. After 18 hours, the cells were harvested and lysed. Subsequently, the amount of expressed protein was evaluated by Western blotting using antibodies against the HA tag. Using FACS and Western blotting, we were able to confirm the translation of mRNA constructs encoding single or multiple smORFs, and the stability of the fusion proteins encoded by them, via CTLA-4 or HA staining.

[0670] [Example 10: In vitro immunogenicity of mRNA constructs encoding multiple smORFs and / or TSAs] Prepare mRNA constructs encoding multiple smORFs and / or TSAs as described above (see R12300 and R12301 in Example 8). Alternatively, prepare variants in which the HA tag is replaced with a helper epitope, for example, as follows: Variant R12300 (mRNA encoding two smORFs expressed in melanoma): 5'UTR-SP(CTLA4)-linker(GS4)-smORF1-linker(GS4)-smORF2-linker(GS4)-helper epitope-linker(GS4)-TMD / TMCD(CTLA4)-3'UTR-polyA Variant R12301 (mRNA encoding six smORFs expressed in NSCLC): 5'UTR-SP(CTLA4)-linker(GS4)-smORF2-linker(GS4)-smORF3-linker(GS4)-smORF5-linker(GS4)-smORF6-linker(GS4)-smORF7-linker(GS4)-smORF4-linker(GS4)-helper epitope-linker(GS4)-TMD / TMCD(CTLA4)-3'UTR-polyA.

[0671] The in vitro immunogenicity of the mRNA construct is analyzed using an assay similar to that described in Example 6. The first stimulation cycle is performed using mRNA, and the second stimulation cycle uses the corresponding peptide.

[0672] [Example 11: In vivo immunogenicity of mRNA construct encoding smORF] To analyze the in vivo immunogenicity of the mRNA constructs encoding smORF, R12300 and R12301 constructs were prepared according to the method described in Example 8 and tested as follows.

[0673] For vaccination, the construct was applied to female CB6F1 hybrid mice. Administration was performed intramuscularly (in the tibialis muscle) on days 0, 7, and 14, with 5 μg of LNP-formulated mRNA (R12300, R12301, or PpLuc control mRNA R8730).

[0674] Twenty-one days after the initial mRNA administration, mice were sacrificially harvested from their spleens for further analysis. Cancer antigen-specific cellular responses in the splenic cell samples obtained in this process were measured as antigen-specific T cell activation. This was analyzed by intracellular cytokine staining and subsequent flow cytometry according to a standard protocol. The protocol was as follows: Isolated splenic cells were individually restimulated with a peptide library for each smORF (a 15-mer library covering the entire antigen), cultured for 6 hours in the presence of anti-CD28 antibody (BD Biosciences, San Jose, USA) at a final peptide concentration of 1 μg / ml, and then GolgiPlug (BD Biosciences) was added after 1 hour. Unstimulated splenic cells were treated similarly, but supplemented with DMSO instead of the peptide cocktail. As additional controls, splenocytes stimulated with PMA / ionomycin (without anti-CD28; PMA and ionomycin were obtained from Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) (positive control) and splenocytes not stained with fluorescent dye-labeled antibodies (negative control) were used. After stimulation, the surface and intracellular environment of splenocytes were stained with fluorescent dye-labeled antibodies and analyzed by flow cytometry. CD8 and CD4 responses are shown as IFN-γ / TNF-α double-positive CD8 and CD4 T cells.

[0675] [result] Representative data for construct R12300 are shown in Figure 18. These data indicate that mRNA R12300 elicited a strong CD8 (top) and CD4 (bottom) response to the lncCLEC2D91412616 smORF. Representative data for T cell responses to construct R12301 and NKZW-smORFs (KCNMB2AS14633753, lncNTF35511951417, lncWDR722415641696, lncZC3H86163144) are shown in Figure 19. These data indicate that R12301 elicited a strong CD8 (top) and ...

Claims

1. An artificial nucleic acid comprising at least one coding sequence encoding at least one tumor antigen, The tumor antigen comprises or consists of at least one antigenic peptide selected from a peptide or protein encoded by a long non-coding RNA (lncRNA), or an immunogenic fragment thereof or a variant thereof. Artificial nucleic acid.

2. The at least one antigenic peptide is A peptide or protein encoded by a long non-coding RNA (lncRNA) 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, or Any of those immunogenic fragments or any of those variants, An artificial nucleic acid according to claim 1, selected from the following.

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

4. The at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA) is At least one amino acid sequence that is identical to any one of sequence numbers 124-254, 546-650, or 756-763, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, The artificial nucleic acid according to claims 1 to 3, comprising or consisting of the above.

5. i) at least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-NTF3-5:5, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 169-226, 604-629, or 760, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, A coding array containing or consisting of; or, ii) At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-WDR72-2:4, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 124-146, 546-571, 756, or 757, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, A coding array containing or consisting of; or, iii) At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-TRPC5-3:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 227-237, 630-642, or 761, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, A coding array containing or consisting of; or, iv) at least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-CLEC2D-9:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 238-243, 643-650, or 762, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, A coding array containing or consisting of; or, v) At least one antigenic peptide selected from the peptides or proteins encoded by KCNMB2-AS1:4, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 147-161, 572-589, or 758, or at least 80% identical to any one of these sequences, Any of those immunogenic fragments or any of those variants, An antigenic peptide containing or consisting of; or, vi) at least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-ZC3H8-6:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 162-168, 590-603, or 759, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, A coding array containing or consisting of; or, vii) At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from peptides or proteins encoded by INC00893:25, Here, the antigenic peptide is At least one amino acid sequence that is identical to any one of sequence numbers 244-254 or 763, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those variants, A coding array that includes or consists of The artificial nucleic acid according to claims 1 to 4, including the above.

6. i) at least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-NTF3-5:5, Here, the antigenic peptide is At least one amino acid sequence that is identical to, or at least 80% identical to, SEQ ID NO: 760, or The immunogenic fragment or its variant, A coding array containing or consisting of; or, ii) At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-WDR72-2:4, Here, the antigenic peptide is At least one amino acid sequence that is identical to, or at least 80% identical to, SEQ ID NO: 756, or The immunogenic fragment or its variant, A coding array containing or consisting of; or, iii) At least one antigenic peptide selected from the peptide or protein encoded by KCNMB2-AS1:4, Here, the antigenic peptide is At least one amino acid sequence that is identical to, or at least 80% identical to, SEQ ID NO: 758, or The immunogenic fragment or its variant, An antigenic peptide containing or consisting of; or, iv) at least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-ZC3H8-6:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to, or at least 80% identical to, SEQ ID NO: 759, or The immunogenic fragment or its variant, A coding array containing or consisting of; or, v) At least one coding sequence encoding at least one tumor antigen comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-CLEC2D-9:1, Here, the antigenic peptide is At least one amino acid sequence that is identical to, or at least 80% identical to, SEQ ID NO: 762, or The immunogenic fragment or its variant, A coding array that includes or consists of The artificial nucleic acid according to claims 1 to 5, including the above.

7. The at least one coding sequence comprises a nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins encoded by lncRNA, preferably a GC-optimized nucleic acid sequence. Here, the nucleic acid sequence is It is identical to any one of sequence numbers 318-448, 651-755, or 764-771, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or It is a fragment of any of those or a variant of any of those. The artificial nucleic acid according to claims 1 to 6.

8. i) at least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from the peptide or protein encoded by lnc-NTF3-5:5, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 363-420, 709-734, or 768, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; or, ii) at least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from the peptide or protein encoded by lnc-WDR72-2:4, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 318-340, 651-676, 764, or 765, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; or, iii) At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-TRPC5-3:1, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 421-431, 735-747, or 769, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; or, iv) at least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, which comprises at least one antigenic peptide selected from the peptide or protein encoded by lnc-CLEC2D-9:1, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 432-437, 748-755, or 770, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; or, v) At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, comprising at least one antigenic peptide selected from the peptide or protein encoded by KCNMB2-AS1:4, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 341-355, 677-694, or 766, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; or, vi) at least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, comprising at least one antigenic peptide selected from the peptide or protein encoded by lnc-ZC3H8-6:1, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 356-362, 695-708, or 767, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; or, vii) At least one coding sequence comprising a nucleic acid sequence encoding at least one tumor antigen, comprising at least one antigenic peptide selected from peptides or proteins encoded by LINC00893:25, Here, the nucleic acid sequence is It is identical to any one of sequence numbers 438-448 or 771, or it is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or A coding array which is any of those fragments or any of those variants; The artificial nucleic acid according to claims 1 to 7, including the above.

9. The artificial nucleic acid according to any one of claims 1 to 8, wherein the at least one coding sequence codes for 2 to 10 different tumor antigens, preferably 2 to 5 different tumor antigens.

10. The artificial nucleic acid according to any one of claims 1 to 9, wherein the at least one coding sequence encodes at least one additional amino acid sequence selected from at least one immune response activation signaling protein, at least one T helper epitope, and / or at least one signal peptide.

11. The artificial nucleic acid according to any one of claims 1 to 10, wherein the at least one coding sequence is a codon-modified coding sequence, preferably, the codon-modified coding sequence is selected from a C-maximizing coding sequence, a CAI-maximizing coding sequence, a coding sequence adapted for human codon use, 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 claims 1 to 12, wherein the nucleic acid comprises at least one untranslated region (UTR), preferably the untranslated region being selected from at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR.

14. The aforementioned at least one heterogeneous 3'-UTR, PSMB3, ALB7, α-globin, β-globin, ANXA4, CASP1, COX6B1, FIG4, GNAS, NDUFA1, RPS9, SLC7A3, or TUBB4B, or Any one homolog, fragment, or variant of those genes, It contains, or consists of, a nucleic acid sequence derived from the 3'-UTR of a gene selected from, Preferably, here, the heterogeneous 3'-UTR is Nucleic acid sequences that are identical to sequence numbers 66-95, 112-123, or are identical to them by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, The artificial nucleic acid according to claim 13, comprising or consisting of the following.

15. The aforementioned at least one heterogeneous 5'-UTR, HSD17B4, RPL32, AIG1, α-globin, ASAH1, ATP5A1, COX6C, DPYSL2, MDR, MP68, NDUFA4, NOSIP, RPL31, RPL35A, SLC7A3, TUBB4B, or UBQLN2, or Any one homolog, fragment, or variant of those genes, It contains, or consists of, a nucleic acid sequence derived from the 5'-UTR of a gene selected from, Preferably, the heterogeneous 5'-UTR is, Nucleic acid sequences that are identical to sequence numbers 12-45, 64, and 65, or are identical to them by at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, The artificial nucleic acid according to claim 13, comprising or consisting of the following.

16. The artificial nucleic acid according to claims 13 to 15, wherein the at least one heterogeneous 5'-UTR is selected from HSD17B4, and the at least one heterogeneous 3'-UTR is selected from PSMB3.

17. The artificial nucleic acid according to any one of claims 1 to 16, wherein the artificial nucleic acid is selected from DNA or RNA, preferably from RNA.

18. The artificial nucleic acid according to any one of claims 1 to 17, wherein the artificial nucleic acid is RNA selected from mRNA, circular RNA, replicon RNA, or viral RNA, and is preferably mRNA.

19. The artificial nucleic acid according to any one of claims 1 to 18, wherein the artificial nucleic acid preferably comprises at least one poly(A) sequence, and preferably, 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, and optionally the nucleotide at the 3' end is adenosine.

22. The artificial nucleic acid, preferably the RNA, comprises at least one histone stem-loop sequence, where the histone stem-loop sequence is A nucleic acid sequence that is identical to either SEQ ID NO: 3 or 4, or identical to either of these by at least 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or Any of those fragments or any of those variants, An artificial nucleic acid according to any one of claims 1 to 21, comprising or consisting of the following.

23. The aforementioned artificial nucleic acid, RNA containing at least one modified nucleotide, Preferably, the RNA contains a modified nucleotide selected from pseudouridine (ψ) or N1-methylpseudridine (m1ψ). More preferably, the RNA contains N1-methylpseudridine (m1ψ). The artificial nucleic acid according to any one of claims 1 to 22.

24. The artificial nucleic acid according to claims 1 to 22, wherein the artificial nucleic acid is RNA that does not contain modified nucleotides.

25. The aforementioned artificial nucleic acid, RNA containing a 5'-cap structure, Preferably, the RNA includes a cap 1 structure or a modified cap 1 structure. The artificial nucleic acid according to any one of claims 1 to 24.

26. The artificial nucleic acid according to any one of claims 1 to 25, wherein the artificial nucleic acid is in vitro transcribed RNA.

27. The artificial nucleic acid, preferably the RNA, preferably contains the following sequence elements in the 5' to 3' direction: A) 5'-cap structure; B) Preferably a 5'-UTR selected from the 5'-UTR of the HSD17B4 gene; C) A coding sequence encoding at least one tumor antigen; D) Preferably a 3'-UTR selected from the 3'-UTR of the PSMB3 gene; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides, The artificial nucleic acid according to any one of claims 1 to 26.

28. The aforementioned artificial nucleic acid is mRNA containing the following sequence elements in the 5' to 3' direction: A) 5'-cap structure; B) A 5'-UTR comprising, or consisting of, a nucleic acid sequence, fragment thereof, or variant thereof, which is identical to, or at least 80% identical to, SEQ ID NO: 13; C) A coding sequence comprising a nucleic acid sequence encoding at least one antigenic peptide selected from peptides or proteins encoded by long non-coding RNA (lncRNA), wherein the nucleic acid sequence is identical to any one of sequence numbers 318-448, 651-755, or 764-771, or is at least 80% identical to any one of these, or is a fragment or variant of any of these; D) A 3'-UTR comprising or consisting of a nucleic acid sequence, fragment thereof, or variant thereof, which is identical to or at least 80% identical to Sequence ID No. 67; E) Optionally, histone stem loops; and, F) Preferably a poly(A) sequence containing about 100 A nucleotides, The artificial nucleic acid according to any one of claims 1 to 27.

29. A pharmaceutical composition comprising at least one artificial nucleic acid, The artificial nucleic acid is an artificial nucleic acid as defined in any one of claims 1 to 28, comprising at least one coding sequence encoding at least one tumor antigen. Pharmaceutical composition.

30. The pharmaceutical composition according to claim 29, comprising a plurality of artificial nucleic acids, each encoding at least one different tumor antigen.

31. The at least one artificial nucleic acid, preferably the RNA, is formulated within at least one cationic or polycationic compound. Here, the at least one cationic or polycationic compound is optionally selected from cationic or polycationic polymers, cationic or polycationic polysaccharides, cationic or polycationic lipids, cationic or polycationic proteins, cationic or polycationic peptides, or any combination thereof. The pharmaceutical composition according to claim 29 or 30.

32. The pharmaceutical composition according to claims 29 to 31, wherein the at least one artificial nucleic acid, preferably RNA, is formulated within a lipid-based carrier.

33. The pharmaceutical composition according to claim 32, wherein the lipid-based carrier is selected from liposomes, lipid nanoparticles, lipoplexes, solid lipid nanoparticles, lipopolyplexes, and / or nanoliposomes.

34. The pharmaceutical composition according to claim 32 or 33, wherein the lipid-based carrier is lipid nanoparticles.

35. The lipid-based carrier preferably contains the LNP, (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 aggregation-inhibiting lipid, A pharmaceutical composition according to claims 32 to 34, comprising:

36. The pharmaceutical composition according to claims 32 to 37, wherein the lipid-based carrier has a Z-average size in the range of about 50 nm to about 200 nm, preferably in the range of about 50 nm to about 150 nm.

37. A pharmaceutical composition or artificial nucleic acid according to any one of claims 1 to 36, wherein the composition or nucleic acid produces an encoded tumor antigen when administered intramuscularly, intratumorally, or intravenously to cells, tissues, or subjects.

38. A pharmaceutical composition or artificial nucleic acid according to any one of claims 1 to 37, wherein when the composition or nucleic acid is administered intramuscularly, intratumorally, or intravenously to cells, tissues, or a target, an encoded tumor antigen is produced, thereby inducing epitope-specific CD8+ T cells in the target.

39. A tumor antigen, or a composition of at least one tumor antigen, wherein the tumor antigen is characterized by any one of claims 1 to 38, preferably any one of claims 2 to 6. More preferably, the tumor antigen is, At least one amino acid sequence that is identical to any one of sequence numbers 124-254, 546-650, or 756-763, or that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of these sequences, or Any of those immunogenic fragments or any of those immunogenic variants, including or consisting of A tumor antigen, or a composition of at least one tumor antigen.

40. A tumor antigen or nucleic acid as defined in any one of claims 1 to 39, or stimulated by thereof Preferably, an antibody, T cell, or TCR prepared for or stimulated by the tumor antigen of claim 39.

41. 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, A combination of at least two, or more, treatment modalities selected from the above.

42. A formulation comprising at least one artificial nucleic acid according to any one of claims 1 to 28, and / or at least one pharmaceutical composition according to any one of claims 29 to 38, and / or at least one tumor antigen or tumor antigen composition according to claim 39, and / or at least one antibody, T cell, or TCR according to claim 40. It optionally contains a liquid vehicle for dissolution, and, Optionally includes technical instructions providing information on the administration and dosage of the ingredients. A kit or parts kit.

43. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or composition of a tumor antigen according to claim 39, an antibody, T cell, or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical.

44. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical for treating or preventing cancer or any cancer-related disease, disorder or condition in a subject.

45. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical for treating or preventing squamous cell carcinoma or any disease, disorder or condition related to squamous cell carcinoma in a subject.

46. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or composition of a tumor antigen according to claim 39, an antibody, T cell or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical for treating or preventing NSCLC or any disease, disorder or condition (particularly squamous cell carcinoma / squamous non-small cell lung cancer) in a subject.

47. The use as a pharmaceutical for treating or preventing NSCLC as described in claim 46, Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is Peptides or proteins encoded by long non-coding RNAs (lncRNAs) selected from lnc-WDR72-2:4, lnc-TRPC5-3:1, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, Those fragments or their variants, Uses a substance containing at least one antigenic peptide selected from the following.

48. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical for treating or preventing HNSCC or any disease, disorder or condition related to HNSCC in a subject.

49. The use as a pharmaceutical for treating or preventing HNSCC as described in claim 48, Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is Peptides or proteins encoded by long non-coding RNAs (lncRNAs) selected from lnc-WDR72-2:4, KCNMB2-AS1:4, lnc-NTF3-5:5, or lnc-ZC3H8-6:1, Those fragments or their variants, Uses a substance containing at least one antigenic peptide selected from the following.

50. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical for treating or preventing melanoma or any disease, disorder or condition related to melanoma in a subject.

51. The use as a pharmaceutical for treating or preventing melanoma, as described in claim 50, Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by a long non-coding RNA (lncRNA) selected from lnc-CLEC2D-9:1, lnc-TRPC5-3:1, or LINC00893:25, Those fragments or their variants, Uses a substance containing at least one antigenic peptide selected from the following.

52. Artificial nucleic acids according to any one of claims 1 to 28, pharmaceutical compositions according to any one of claims 29 to 38, tumor antigens or compositions of tumor antigens according to claim 39, antibodies, T cells or TCRs according to claim 40, combinations according to claim 41, or kits or component kits according to claim 42, for use as pharmaceuticals to treat or prevent adrenocortical carcinoma, urothelial carcinoma of the bladder, invasive breast cancer, cervical cancer (squamous cell carcinoma of the cervix and adenocarcinoma of the cervix), colon adenocarcinoma, renal papillary cell carcinoma, lung adenocarcinoma, lymphoid tumors, diffuse large B cell lymphoma, rectal adenocarcinoma, sarcoma, cutaneous melanoma, thymoma, thyroid cancer, hepatocellular carcinoma, uterine carcinosarcoma, or endometrial cancer of the uterine body, or any disease, disorder or condition related thereto, in the target area. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-ZC3H8-6:1, or The fragment or variant thereof, comprising at least one antigenic peptide selected from, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, or kits or component kits.

53. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell, or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical to treat or prevent bladder urothelial carcinoma, cervical cancer (cervical squamous cell carcinoma and cervical adenocarcinoma), thymoma, or endometrial cancer of the uterine body, or any disease, disorder, or condition related thereto, in a subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by KCNMB2-AS1:4, or The fragment or variant thereof, comprising at least one antigenic peptide selected from, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, or kits or component kits.

54. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell, or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical to treat or prevent ovarian serous cystadenocarcinoma, testicular germ cell tumor, thymoma, or uterine carcinosarcoma, or any disease, disorder, or condition related thereto, in a subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-NTF3-5:5, or The fragment or variant thereof, comprising at least one antigenic peptide selected from, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, or kits or component kits.

55. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell, or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical to treat or prevent bladder urothelial carcinoma, colon adenocarcinoma, esophageal cancer, or gastric adenocarcinoma, or any disease, disorder, or condition related thereto, in a subject. Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-WDR72-2:4, or The fragment or variant thereof, comprising at least one antigenic peptide selected from, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, or kits or component kits.

56. An artificial nucleic acid according to any one of claims 1 to 28, a pharmaceutical composition according to any one of claims 29 to 38, a tumor antigen or tumor antigen composition according to claim 39, an antibody, T cell, or TCR according to claim 40, a combination according to claim 41, or a kit or component kit according to claim 42, for use as a pharmaceutical to treat or prevent cutaneous melanoma, testicular germ cell tumor, or uveal melanoma or any disease, disorder, or condition related thereto in a subject, Here, the artificial nucleic acid, the pharmaceutical composition, the tumor antigen or composition of tumor antigen, the combination, or at least one tumor antigen provided by the kit or component kit is A peptide or protein encoded by lnc-CLEC2D-9:1, or The fragment or variant thereof, comprising at least one antigenic peptide selected from, Artificial nucleic acids, pharmaceutical compositions, tumor antigens or compositions of tumor antigens, antibodies, T cells or TCRs, combinations, or kits or component kits.

57. A method for treating or preventing a disease, disorder, or condition, The method, in this case, includes applying or administering an effective amount of the artificial nucleic acid described in claims 1 to 28, the pharmaceutical composition described in claims 29 to 38, the tumor antigen or tumor antigen composition described in claim 39, the antibody, T cell, or TCR described in claim 40, the combination described in claim 41, or the kit or component kit described in claim 42 to a target that requires it.

58. The method according to claim 57, wherein the method is further characterized by any one of the characteristics of a medical use as defined in claims 43 to 56.

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