Anti-pd-l1 vaccine compositions

Abstract

The present invention relates to a polypeptide comprising or consisting of: - a first sequence consisting of at least 8 contiguous amino acid residues and at most 30 contiguous amino acid residues, said at least 8 contiguous amino acid residues being selected from the sequence extending at amino acid residues 55-67 of the PD-L1 protein and said at most 30 contiguous amino acid residues being selected from the complete sequence of the PD-L1 protein; and / or - a second sequence consisting of at least 8 contiguous amino acid residues and at most 30 contiguous amino acid residues, said at least 8 contiguous amino acid residues being selected from the sequence extending at amino acid residues 85 to 101 of the PD-L1 protein and said at most 30 contiguous amino acid residues being selected from the complete sequence of the PD-L1 protein; and / or - a third sequence consisting of at least 8 contiguous amino acid residues and at most 30 contiguous amino acid residues, said at least 8 contiguous amino acid residues being selected from the sequence extending at amino acid residues 111 to 127 of the PD-L1 protein and said at most 30 contiguous amino acid residues being selected from the complete sequence of the PD-L1 protein; and / or - a fourth sequence consisting of at least 8 contiguous amino acid residues and at most 30 contiguous amino acid residues, said at least 8 contiguous amino acid residues being selected from the sequence extending at amino acid residues 138 to 156 of the PD-L1 protein and said at most 30 contiguous amino acid residues being selected from the complete sequence of the PD-L1 protein; and / or - a fifth sequence consisting of at least 8 contiguous amino acid residues and at most 30 contiguous amino acid residues, said at least 8 contiguous amino acid residues being selected from the sequence extending at amino acid residues 208 to 223 of the PD-L1 protein and said at most 30 contiguous amino acid residues being selected from the complete sequence of the PD-L1 protein.

Description

TECHNICAL FIELD

[0001] The present invention relates to polypeptides useful for eliciting an immune response against PD-L1 protein. BACKGROUND

[0002] It has now been established that the strength of the natural immune response against cancer antigens correlates with a better prognosis of the patient and of various types of neoplasia. Clinical observations supported by a large body of experimental evidence allowed the definition of the concept of cancer immunosurveillance, according to which newly arising tumors are usually eliminated by the immune system (unless the cancer cells have evolved to escape immune detection).

[0003] Anti-cancer immunotherapy - a direct application of the immunosurveillance concept - has experienced an amazing growth and success in the last decade, revolutionizing the clinical management of a wide range of malignancies with a previously dismal prognosis.

[0004] The most advanced development of immunotherapy is represented by monoclonal antibodies, i.e. immune response checkpoint blockers (ICB), which have been very successful in oncology due to their broad activity on multiple types of tumors, their responses in metastatic chemoresistant tumors and the durability of their therapeutic capacity.

[0005] Among the checkpoint blockade strategies, two are the most important (in terms of clinical success so far) are the targeting of monoclonal antibodies specific for Cytotoxic T Lymphocyte-Associated Protein 4 (CTLA-4) and the interaction between Programmed Cell Death Protein 1 (PD-1) and the Programmed Cell Death Protein 1 Ligand (PD-L1). In particular, the inhibition of PD-L1 signaling has been proposed as a means to increase the immunocompetence of T cells for the treatment of cancer (anti-tumor immunity) and also for the treatment of infections (chronic and acute persistent infections).

[0006] So far, four ICBs targeting the PD-1 / PD-L1 axis have been approved by the Food and Drug Administration (FDA) (for a review see, for example, Abdin et al. (2018) Cancers 1032):

[0007] (1) pembrolizumab, an anti-PD-1 monoclonal antibody (mAb), has been approved for patients with unresectable metastatic melanoma or advanced metastatic non-small cell lung cancer (NSCLC) whose tumors express PD-L1;

[0008] (2) nivolumab, an anti-PD-1 monoclonal antibody (mAb), has been approved for unresectable or metastatic melanoma, advanced metastatic NSCLC that has progressed during or after platinum-based chemotherapy, and, in particular, metastatic advanced renal carcinoma; and

[0009] (3) Atezolizumab, an anti-PD-L1 monoclonal antibody (mAb), was recently approved for the treatment of locally advanced or metastatic urothelial carcinoma that is unresponsive to platinum-derived chemotherapy; and

[0010] (4) Avelumab, an anti-PD-L1 monoclonal antibody (mAb), was recently approved for the treatment of metastatic Merkel cell carcinoma.

[0011] In addition, these ICBs, which have already been approved for many indications, may also be considered for the treatment of other forms of cancer in the future.

[0012] In addition, ICBs targeting PD-L1 have been shown to be very effective in melanoma, NSCLC, and renal cell carcinoma.

[0013] However, all products sold or developed by ICBs targeting the PD-1 / PD-L1 axis are monoclonal antibodies, and therefore suffer from the same limitations as other monoclonal antibody therapies: high cost, the need for frequent reapplication, and the development of an immune response to the applied monoclonal antibody.

[0014] Therefore, the purpose of this invention is to overcome these disadvantages. Summary of the Invention

[0015] This invention stems from the inventors’ unexpected demonstration that a polypeptide derived from the extended sequence of amino acid residues 55 to 67, 85 to 101, 111 to 127, 138 to 156 and 208 to 223 of the human PD-L1 protein allows mice to produce antibodies that neutralize the PD-L1 protein.

[0016] Therefore, the present invention relates to polypeptides comprising or composed of the following:

[0017] -

[0018] A first sequence comprising at least 8 consecutive amino acid residues and at most 30 consecutive amino acid residues, wherein the at least 8 consecutive amino acid residues are selected from the sequence extending from amino acid residues 55-67 of the PD-L1 protein, and the at most 30 consecutive amino acid residues are selected from the complete sequence of the PD-L1 protein, or a variant sequence having at least 75% identity with the first sequence; and / or

[0019] - A second sequence comprising at least 8 consecutive amino acid residues and at most 30 consecutive amino acid residues, wherein the at least 8 consecutive amino acid residues are selected from a sequence extending from amino acid residues 85 to 101 of the PD-L1 protein, and the at most 30 consecutive amino acid residues are selected from the complete sequence of the PD-L1 protein, or a variant sequence having at least 75% identity with the second sequence; and / or

[0020] - A third sequence comprising at least 8 consecutive amino acid residues and at most 30 consecutive amino acid residues, wherein the at least 8 consecutive amino acid residues are selected from the sequence extending from amino acid residues 111 to 127 of the PD-L1 protein, and the at most 30 consecutive amino acid residues are selected from the complete sequence of the PD-L1 protein, or a variant sequence having at least 75% identity with the third sequence; and / or

[0021] - A fourth sequence comprising at least 8 consecutive amino acid residues and at most 30 consecutive amino acid residues, wherein the at least 8 consecutive amino acid residues are selected from the sequence extending from amino acid residues 138 to 156 of the PD-L1 protein, and the at most 30 consecutive amino acid residues are selected from the complete sequence of the PD-L1 protein, or a variant sequence having at least 75% identity with the fourth sequence; and / or

[0022] - A fifth sequence comprising at least 8 consecutive amino acid residues and at most 30 consecutive amino acid residues, wherein the at least 8 consecutive amino acid residues are selected from the sequence extending from amino acid residues 208 to 223 of the PD-L1 protein, and the at most 30 consecutive amino acid residues are selected from the complete sequence of the PD-L1 protein, or a variant sequence having at least 75% identity with the fifth sequence; and / or

[0023] The premise is that the polypeptide is different from the PD-L1 protein and it is not composed of a portion of the PD-L1 protein consisting of more than 30 consecutive amino acid residues, and

[0024] The polypeptide, composed of variant sequences of the first, second, third, fourth, and fifth sequences, allows for the initiation of an immune response against the PD-L1 protein.

[0025] In a preferred embodiment, the present invention more specifically relates to a polypeptide comprising or composed of the following:

[0026] - A first sequence consisting of SEQ ID NO: 1, 51, 52 or 53, or a variant sequence having at least 75% identity with the first sequence; and / or

[0027] - A second sequence consisting of SEQ ID NO: 2, 54 or 55, or a variant sequence having at least 75% identity with the second sequence; and / or

[0028] - A third sequence consisting of SEQ ID NO: 3, 56, 57, 58 or 59, or a variant sequence having at least 75% identity with the third sequence; and / or

[0029] - A fourth sequence consisting of SEQ ID NO: 4 or 60, or a variant sequence having at least 75% identity with the fourth sequence; and / or

[0030] - A fifth sequence consisting of SEQ ID NO: 5, 61 or 62, or a variant sequence having at least 75% identity with the fifth sequence;

[0031] The premise is that the polypeptide composed of variant sequences of the first, second, third, fourth, and fifth sequences, respectively, allows the initiation of an immune response against the PD-L1 protein.

[0032] The present invention also relates to nucleic acids encoding polypeptides as defined above or nucleic acids thereof with complementary sequences.

[0033] This invention also relates to:

[0034] - at least one polypeptide as defined above, or

[0035] -At least one nucleic acid as defined above,

[0036] It is used as a medicine, particularly a vaccine. In one specific embodiment of the invention, the medicine, particularly the vaccine, as defined above, further comprises at least one other compound intended to prevent or treat diseases, cancers, or infectious diseases associated with or attributable to the expression of the PD-L1 protein or the PD-1 protein.

[0037] This invention also relates to pharmaceutical compositions, particularly vaccines, comprising an active substance:

[0038] - at least one polypeptide as defined above, or

[0039] -At least one nucleic acid as defined above,

[0040] Optionally, it may be combined with at least one pharmaceutically acceptable carrier. In one specific embodiment of the invention, the pharmaceutical composition, particularly a vaccine, as defined above, further comprises at least one other compound intended to prevent or treat diseases, cancers, or infectious diseases associated with or attributable to the expression of the PD-L1 protein.

[0041] This invention also relates to the use of polypeptides as defined above in the preparation of antibodies, antibody fragments or aptamers.

[0042] The present invention also relates to a method for preparing antibodies, antibody fragments or aptamers, the method comprising the step of administering a polypeptide as defined above to an antibody-producing organism or the step of selecting an antibody, antibody fragment or aptamer that binds to the polypeptide based on affinity.

[0043] The present invention also relates to anti-PD-L1 antibodies, antibody fragments or aptamers specifically targeting the above-mentioned polypeptides, provided that the polypeptide does not contain more than two amino acid residues other than the first, second, third, fourth or fifth sequences or their respective variant sequences.

[0044] The present invention also relates to antibodies, antibody fragments, or aptamers as defined above, used as pharmaceuticals. In one specific embodiment of the invention, the pharmaceutical as defined above further comprises at least one other compound intended to prevent or treat diseases, cancers, or infectious diseases associated with or attributable to the expression of PD-L1 protein or PD-1 protein.

[0045] The present invention also relates to pharmaceutical compositions comprising, as an active ingredient, an antibody, antibody fragment, or aptamer as defined above, optionally combined with a pharmaceutically acceptable carrier. In one specific embodiment of the invention, the pharmaceutical composition as defined above further comprises at least one other compound intended to prevent or treat diseases, cancers, or infectious diseases associated with or attributable to the expression of PD-L1 protein or PD-1 protein.

[0046] The present invention also relates to a method for inducing an immune response against the PD-L1 protein in an individual using a polypeptide, nucleic acid, or pharmaceutical composition as defined above. In one specific embodiment of the invention, the polypeptide, nucleic acid, or pharmaceutical composition is used in combination with at least one other compound that can be used to induce an immune response against the PD-L1 protein.

[0047] The present invention also relates to a method for inducing an immune response against the PD-L1 protein in an individual, comprising administering to the individual an effective amount of a polypeptide, a nucleic acid, or a pharmaceutical composition as defined above. In one specific embodiment of the invention, the polypeptide, nucleic acid, or pharmaceutical composition is administered in combination with at least one other compound that can be used to induce an immune response against the PD-L1 protein.

[0048] This invention also relates to the use of polypeptides or nucleic acids as defined above in the preparation of medicaments intended to elicit an immune response against the PD-L1 protein in an individual. In one specific embodiment of the invention, the medicament further comprises at least one other compound that can be used to elicit an immune response against the PD-L1 protein.

[0049] This invention also relates to peptides, nucleic acids, pharmaceutical compositions, or antibodies, antibody fragments, or aptamers as defined above, for use in an individual to prevent or treat diseases associated with or attributable to the expression of PD-1 or PD-L1 proteins. In one specific embodiment of the invention, the peptide, nucleic acid, pharmaceutical composition, or antibody, antibody fragment, or aptamer is combined with at least one other therapy intended to prevent or treat diseases, cancers, or infectious diseases associated with or attributable to the expression of PD-1 or PD-L1 proteins.

[0050] The present invention also relates to methods for preventing or treating diseases in an individual that are related to or attributable to the PD-L1 protein, comprising administering to the individual an effective amount of a polypeptide, nucleic acid, pharmaceutical composition, or antibody, antibody fragment, or aptamer as defined above. In one specific embodiment of the invention, the method includes at least one other therapy aimed at preventing or treating diseases, cancers, or infectious diseases associated with the expression of the PD-1 protein or PD-L1 protein.

[0051] This invention also relates to the use of polypeptides, nucleic acids, or antibodies, antibody fragments, or aptamers as defined above for the preparation of medicaments intended to prevent or treat diseases in an individual that are associated with or attributable to the PD-L1 protein. In one specific embodiment of the invention, the medicament comprises at least one other compound intended to prevent diseases, cancers, or infectious diseases associated with or attributable to the expression of the PD-L1 protein.

[0052] This invention also relates to products containing the following:

[0053] - As defined above, polypeptides or nucleic acids, and

[0054] - At least one other compound, said compound being intended to prevent or treat diseases, cancers, or infectious diseases related to or attributable to the PD-L1 protein.

[0055] As a combination product, it is intended to be used simultaneously, separately, or alternately in time to prevent or treat diseases, cancers, or infectious diseases in an individual that are associated with or attributable to the expression of PD-1 or PD-L1 proteins. Detailed Implementation

[0056] First, the term "contains" means "includes," "includes," or "contains," meaning that when an object "includes" one or more elements, other elements besides those mentioned can also be included in the object. Conversely, the expression "composed of" means "consisting of," meaning that when an object "composes of" one or more elements, the object cannot contain other elements besides those mentioned.

[0057] Polypeptide

[0058] Definition of PD-L1 protein

[0059] PD-L1 protein is a ligand protein of PD-1 protein, also known as differentiation cluster 274 (CD274), which is well known to those skilled in the art.

[0060] Preferred types and sequences of PD-L1 proteins

[0061] Preferably, the PD-L1 protein according to the present invention is selected from human PD-L1 protein, mouse PD-L1 protein, monkey PD-L1 protein, horse PD-L1 protein, bovine PD-L1 protein, porcine PD-L1 protein, sheep PD-L1 protein, goat PD-L1 protein, camel PD-L1 protein, dromedary camel PD-L1 protein, dog PD-L1 protein, and cat PD-L1 protein. Particularly preferably, the PD-L1 protein is human PD-L1 protein.

[0062] Preferably:

[0063] - Human PD-L1 protein (hPD-L1), as described in UniProt / Swisspro database number Q9NZQ7, and composed of SEQ ID NO: 6,

[0064] -Monkey PD-L1 protein, as described in Genbank database accession NP_001077358.1, and composed of SEQ ID NO: 7,

[0065] - Mouse PD-L1 protein (mPD-1), as described in UniProt / Swisspro database number Q9EP73, and composed of SEQ ID NO: 8,

[0066] - Horse PD-L1 protein, as described in Genbank database accession XP_001492892.1, and composed of SEQ ID NO: 9,

[0067] - Bovine PD-L1 protein, as described in Genbank database accession NP_001156884.1, and composed of SEQ ID NO: 10,

[0068] - Porcine PD-L1, as described in UniProt / Swisspro database number Q4QTK1, and composed of SEQ ID NO: 11,

[0069] - Sheep PD-L1 protein, as described in Genbank database accession XP_011980010.1, and composed of SEQ ID NO: 12,

[0070] - Goat PD-L1 protein, as described in Genbank database number XP_005683750.1, and composed of SEQ ID NO: 13.

[0071] - Camel PD-L1 protein, as described in Genbank database numbers XP_014416021.1 or XP_010958932.1, and consisting of SEQ ID NO: 14 or 15.

[0072] - The PD-L1 protein from dromedary camels, as described in Genbank database number XP_010991731.1, and consisting of SEQ ID NO: 16.

[0073] -Canine PD-L1 protein, as described in UniProt / Swisspro database under number E2RKZ5, and composed of SEQ ID NO: 17.

[0074] - The feline PD-L1 protein, as described in Genbank database number XP_006939101.1, and composed of SEQ ID NO: 18.

[0075] Amino acid residue numbering

[0076] As understood herein, the amino acid residues of the PD-L1 protein are numbered starting from the first amino acid residue, typically methionine (M), forming the N-terminus of the complete PD-L1 encoded by the open reading frame of the PD-L1 gene, i.e., including its signal peptide. Furthermore, the amino acid residue numbering of the PD-L1 protein used herein is defined with reference to the human PD-L1 protein. Therefore, it is readily apparent to those skilled in the art that the amino acid residues of the PD-L1 protein corresponding to the position number referenced according to the invention can be determined by comparing the sequence of the PD-L1 protein for which the position number is desired to be determined with the sequence of the human PD-L1 protein, particularly SEQ ID NO: 6, to optimize the percentage of identity between the two aligned sequences, and then identifying the amino acid residue corresponding to the sought position number as the residue aligned with the amino acid residue of the human PD-L1 protein sequence having that position number.

[0077] First, second, third, fourth and fifth sequences

[0078] Preferably, the first, second, third, fourth, and fifth sequences are each composed of at least 8, 9, 10, 11, and 12 consecutive amino acid residues selected from sequences extended from amino acid residues 55 to 67, 85 to 101, 111 to 127, 138 to 156, and 208 to 223 of the PD-L1 protein, or are each composed of sequences extended from amino acid residues 55 to 67, 85 to 111, 101 to 127, 138 to 156, and 208 to 223 of the PD-L1 protein.

[0079] Preferably, the first, second, third, fourth, and fifth sequences according to the present invention are each composed of up to 29, 28, 27, 26, 25, 24, 23, 22, 21, and 20 consecutive amino acid residues selected from the complete sequence of the PD-L1 protein, or are each composed of up to 55 to 67, 85 to 101, 111 to 127, 138 to 156, and 208 to 223 amino acid residues of the PD-L1 protein.

[0080] Preferably:

[0081] The first sequence according to the present invention consists of SEQ ID NO: 1, 51, 52 or 53.

[0082] The second sequence according to the present invention consists of SEQ ID NO: 2, 54 or 55.

[0083] According to the present invention, the third sequence consists of SEQ ID NO: 3, 56, 57, 58 or 59.

[0084] The fourth sequence according to the invention consists of SEQ ID NO: 4 or 60, and

[0085] The fifth sequence according to the present invention consists of SEQ ID NO: 5, 61 or 62.

[0086] The table below shows the amino acid residues of the human PD-L1 protein corresponding to these sequences:

[0087] Sequence SEQ ID NO: hPD-L1 amino acid residues VYWEMEDKNIIQF 1 55-67 YWEMEDKNIIQF 51 56-67 VYWEMEDKNIIQ 52 55-66 YWEMEDKNIIQ 53 56-66 ARLLKDQLSLGNAALQI 2 85-101 RLLKDQLSLGNAALQI 54 86-101 ARLLKDQLSLGNAALQ 55 85-100 VYRCMISYGGADYKRIT 3 111-127 YRCMISYGGADYKRIT 56 112-127 VYRCMISYGGADYKRI 57 111-126 RCMISYGGADY 58 113-123 YRCMISYGGADYKRI 59 112-126 NQRILVVDPVTSEHELTCQ 4 138-156 QRILVVDPVTSEHELTC 60 139-155 YCTFRRLDPEENHTAE 5 208-223 YCTFRRLDPEENHTA 61 208-222 CTFRRLDPEENHTA 62 209-222

[0088] Variant Sequence

[0089] The variant sequences according to the invention that have at least 75% identity with one of the first, second, third, fourth and fifth sequences described above, preferably have at least 80%, 85%, 90%, 95% or 98% identity with one of the first, second, third, fourth and fifth sequences described above.

[0090] As understood in this paper, the percentage of identity between two peptide sequences can be determined by performing an optimal alignment across the entire length of the sequences. This is done by identifying the number of aligned amino acids that are identical in each sequence and dividing that number by the total number of amino acids in the longer of the two sequences. The optimal alignment is the one that yields the highest percentage of identity between the two sequences.

[0091] Preferably, the variant sequence according to the invention has at least 75%, 80%, 85%, 90%, 95% or 98% identity with SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 or 62.

[0092] The variant sequences according to the invention cause the polypeptides composed of the variant sequences to allow the initiation of an immune response against the PD-L1 protein; that is, the administration of such a peptide, optionally cyclized by forming at least one intercysteine ​​disulfide bond, and if necessary, by adding one or two cysteine ​​residues to the peptide, and / or at its N-terminus and / or its C-terminus, optionally binds to a carrier molecule of an animal (e.g., mouse, rat, or rabbit), particularly a carrier protein such as KLH (hemocyanin), to induce the production of antibodies against PD-L1, particularly against the PD-L1 protein of the same species to which the sequence having the highest percentage of identity with the variant sequence belongs. Those skilled in the art will readily know how to determine whether an antibody is against PD-L1, particularly by performing an ELISA test. Preferably, the antibodies induced by the administration of the peptide are blocking or neutralizing agents, that is, they prevent the PD-L1 protein from exerting all or part of its activity, particularly at least 10%, 25%, 50%, or 75%, as measured, for example, in vitro. As understood herein, PD-L1 activity is preferably associated with the binding of the PD-1 protein, which can be measured as in Example 2.

[0093] Preferably, the variant sequence according to the present invention is selected from:

[0094] -VYRSMISYGGADYKRIT(SEQ ID NO:19),

[0095] -YRSMISYGGADYKRI(SEQ ID NO:63)

[0096] -NQRILVVDPVTSEHELTSQ(SEQ ID NO:20),

[0097] -YSTFRLDPEENHTAE(SEQ ID NO:21),

[0098] - YSTFRRLDPEENHTA (SEQ ID NO: 64).

[0099] VYRSMISYGGADYKRIT (SEQ ID NO:19) is derived from VYRCMISYGGADYKRIT (SEQ ID NO:3) by replacing the cysteine ​​(C) at the fourth position with serine (S).

[0100] YRSMISYGGADYKRI (SEQ ID NO:63) is derived from YRCMISYGGADYKRI (SEQ ID NO:59) by replacing the cysteine ​​(C) at the third position with serine (S).

[0101] NQRILVVDPVTSEHELTSQ (SEQ ID NO:20) is derived from NQRILVVDPVTSEHELTCQ (SEQ ID NO:4) by replacing the cysteine ​​(C) at the penultimate position with serine (S).

[0102] YSTFRRLDPEENHTAE (SEQ ID NO:21) is derived from YCTFRRLDPEENHTAE (SEQ ID NO:5) by replacing the cysteine ​​(C) at the second position with serine (S).

[0103] YSTFRRLDPEENHTA (SEQ ID NO:64) is derived from YCTFRRLDPEENHTA (SEQ ID NO:61) by replacing the cysteine ​​(C) at the second position with serine (S).

[0104] length of peptide

[0105] The polypeptide according to the invention preferably comprises up to 200, 150, 100, 90, 80, 70, 60, 50, 40, or 30 amino acid residues. It differs from the PD-L1 protein and is not composed of a portion of the PD-L1 protein containing more than 30 consecutive amino acid residues. As those skilled in the art will understand, this does not preclude it from being composed of two or more portions of the PD-L1 protein containing up to 30 consecutive amino acid residues, provided that these portions are not arranged by recombining portions of the PD-L1 protein containing more than 30 consecutive amino acid residues.

[0106] It will be apparent to those skilled in the art that the polypeptide according to the invention may include multiple repeats, for example, 2, 3, 4, 5, 10 or 20 repeats of the first, second, third, fourth and fifth sequences and variant sequences according to the invention.

[0107] Sequences other than the first, second, third, fourth, and fifth sequences and their variants.

[0108] Furthermore, the polypeptides according to the invention may also include additional sequences not derived from the PD-L1 protein.

[0109] These additional sequences can specifically bring about physicochemical properties, allowing the polypeptides according to the invention to have improved structural performance or improved solubility compared to similar polypeptides that do not include these additional sequences.

[0110] The additional sequence may also include one or more peptide linker sequences, i.e., adaptor peptides, specifically for binding to carrier molecules. Such peptide linker sequences typically contain 1 to 10, particularly 4 to 6, amino acid residues.

[0111] Furthermore, these sequences are not derived from the PD-L1 protein and may include epitopes belonging to other proteins, thereby allowing the initiation or generation of immune responses against these other proteins.

[0112] In addition, the polypeptides according to the invention may contain sequences of exogenous T epitopes, preferably universal, which allows for enhanced immunogenicity of the polypeptides according to the invention.

[0113] As specifically described in international application WO 05 / 117983, the polypeptide according to the invention may also contain at least one sequence of a carrier protein, such as a virus-type particle (VLP).

[0114] Peptide cyclization

[0115] The polypeptide according to the invention can be in linear or cyclized form. Preferably, the polypeptide according to the invention is in cyclized form. The cyclization can be of any type known to those skilled in the art.

[0116] The choice of cyclization strategy according to the invention can specifically take into account the optimal antigenic expression of the epitopes contained in the polypeptide according to the invention, and involves only a portion of the polypeptide (cyclization with respect to the sequence). Thus, as understood herein, when the polypeptide according to the invention is in cyclized form, only a portion of the polypeptide can be contained in the ring, while the remainder of the polypeptide is in linear form.

[0117] Depending on the functional groups present in the polypeptide, this cyclization can proceed in several different ways, for example: from its C-terminus to the N-terminus, from its N-terminus to the side chain, from the side chain to its C-terminus, or even between two side chains. Among the various methods of polypeptide cyclization, lactamation, lactone formation, or disulfide bond formation can be listed. In particular, in the formation of disulfide bonds between cysteine ​​residues, i.e., between the -SH radicals of two cysteine ​​residues, the cysteine ​​residues may already be present in the variant sequences according to the invention or in the first, second, third, fourth, and fifth sequences according to the invention, or may be added to these sequences and to their N-termini and / or C-termini.

[0118] Post-translational modification, amino acid analogs

[0119] Furthermore, the polypeptides according to the invention may include post-translational modifications, such as glycosylation, methylation, acylation, particularly via fatty acids, or phosphorylation. Specifically, the N-terminus of the polypeptides according to the invention may be acetylated, and the C-terminus may be modified by amidation.

[0120] The polypeptides according to the invention may also contain one or more amino acid analogs or derivatives, non-natural or non-standard amino acids, particularly ortholeucine (Nle).

[0121] carrier molecules

[0122] Preferably, the polypeptide according to the invention is covalently linked or bound to a carrier molecule, particularly a carrier protein.

[0123] Specifically, the carrier molecule can be keyhole hemocyanin (KLH) protein, hepatitis B surface antigen (HBsAg), bovine serum albumin (BSA), tetanus toxin (TT), and diphtheria toxin (DT).

[0124] The diphtheria toxin (DT) according to the present invention is preferably selected from CRM 197, CRM 176, CRM 228, CRM 45, CRM 9, CRM 102, CRM 103, and CRM 107.

[0125] Particularly preferred is the carrier molecule CRM 197.

[0126] The polypeptides according to the invention can be bound to carrier molecules, particularly carrier proteins, using heterobifunctional coupling agents, such as N-γ-maleimide butyryl-N-hydroxysuccinimide ester (GMBS) and sulfonated GMBS derivatives, m-maleimide benzoyl-n-hydroxysuccinimide ester (MBS) and sulfonated MBS derivatives, succinimide 4-(N-maleimide methyl)cyclohexane-1-carboxylic acid ester (SMCC), carbodiimide, diazonium benzidine (BDB), or glutaraldehyde.

[0127] When using GMBS, MBS, or SMCC, they are preferably linked to cysteine ​​(C). If cysteine ​​is absent in the first, second, third, fourth, or fifth sequence, a variant sequence according to the invention can be added, particularly at its N-terminus or C-terminus. Furthermore, according to the invention, when cysteine ​​is present at an undesirable position in the first, second, third, fourth, or fifth sequence, a variant sequence in which cysteine ​​is substituted by another amino acid, such as serine, can be implemented instead, as described for SEQ ID NO:19, 20, 21, 63, and 64 relative to SEQ ID NO:3, 4, 5, 59, and 61, respectively.

[0128] When using BDB, it is preferably linked to tyrosine (Y). If tyrosine is absent in the first, second, third, fourth, or fifth sequence, a variant sequence according to the invention can be added, particularly at its N-terminus or C-terminus. Furthermore, according to the invention, when tyrosine is present at an undesirable position in the first, second, third, fourth, or fifth sequence, a variant sequence in which tyrosine is substituted by another amino acid, such as phenylalanine (F), can be implemented instead.

[0129] Furthermore, the binding of the polypeptide of the present invention to a carrier molecule, particularly a carrier protein, can also be achieved via a peptide linker or adapter peptide, wherein one side of the peptide linker or adapter peptide is bound to the polypeptide according to the present invention and the other side is optionally bound to the carrier molecule via a heterobifunctional coupling agent as defined above. Such peptide links typically contain 1 to 10, particularly 4 to 6, amino acid residues.

[0130] Very particularly preferred, the polypeptide according to the invention is linked to the CRM 197 carrier protein according to a construct represented by the formula selected from:

[0131]

[0132]

[0133]

[0134] in:

[0135] -CRM 197 indicates a carrier protein.

[0136] -GMB represents N-γ-maleimide butyryl.

[0137] -Acetyl+ indicates that the N-terminus is acetylated.

[0138] -amide+ indicates that the C-terminus is amidated.

[0139] - cyclo() indicates a lactam-type cyclization between the side chains of the C-terminal and N-terminal amino acid residues.

[0140] - cycloSS() indicates disulfide cyclization between the thiol groups of cysteine ​​residues at the C-terminus and N-terminus.

[0141] - Square brackets ([X]n) indicate that one or more polypeptides are linked to a carrier protein, and

[0142] - The underlined portion represents a polypeptide according to the invention, whose SEQ ID NO is indicated in the right column.

[0143] Therefore, very particularly preferably, the polypeptide according to the invention consists of a sequence selected from SEQ ID NO: 22 to 50.

[0144] Preparation of peptides

[0145] The polypeptide according to the invention can be prepared by any method known in the prior art, particularly by chemical synthesis. It can also be prepared by expressing the nucleic acid according to the invention in eukaryotic or prokaryotic cells.

[0146] Peptide activity

[0147] If necessary, the polypeptide according to the invention binds to a carrier molecule. This polypeptide is immunogenic, meaning it can trigger or induce an immune response, particularly of a humoral type, i.e., the individual to which it is administered, particularly a mammalian individual, produces antibodies. Specifically, the polypeptide according to the invention allows the induction of an immune response against the PD-L1 protein, particularly anti-PD-L1 antibodies, preferably anti-PD-L1 antibody blockers or neutralizers, i.e., those that prevent the PD-L1 protein from exerting all or part of its activity, particularly, for example, at least 10%, 25%, 50%, or 75% of its activity as measured in vitro. As understood herein, the activity of PD-L1 is preferably bound to the PD-1 protein, which can be measured as shown in Example 2 below.

[0148] Nucleic acid

[0149] The nucleic acid according to the invention is RNA or DNA, preferably DNA. Preferably, the nucleic acid according to the invention is operatively bound to prokaryotic and / or eukaryotic promoter sequences, particularly those of mammals or viruses. Furthermore, the nucleic acid according to the invention can be contained in a vector, such as a plasmid or virus.

[0150] Antibodies, antibody fragments, and aptamers

[0151] When the antibodies, antibody fragments, and aptamers according to the invention substantially do not bind to another polypeptide (not including the polypeptides defined above), the antibodies, antibody fragments, and aptamers are considered to specifically target the polypeptides as defined above, thereby allowing the antibodies, antibody fragments, and aptamers according to the invention to bind to the polypeptides they specifically target.

[0152] The antibodies according to the invention can be polyclonal or monoclonal, preferably monoclonal. Furthermore, as understood herein, an "antibody fragment" comprises at least one antigen-binding portion of the antibody, derived, in particular, of the Fab type, Fab' type, F(ab')2 type, an Fv stabilized by a disulfide bond (dsFv), a dimerized (biantibody), trimerized, tetramerized, or pentamerized V region, a single-chain Fv (scFv), and a complementary region determining the complementary region (CDR).

[0153] Antibodies can be from any species, particularly humans, mice, rats, rabbits, or camels. Furthermore, when they are non-human antibodies, they can be humanized, meaning that the constant portions of these antibodies are partially or completely replaced by the corresponding human constant portions.

[0154] Antibodies according to the invention can be obtained by immunizing animals with the polypeptides according to the invention, using techniques known to those skilled in the art.

[0155] As used herein, an aptamer is a nucleic acid, particularly RNA, capable of specifically binding to molecular targets such as proteins. Aptamers can be obtained, in particular, from the peptides of this invention using the SELEX technique known to those skilled in the art.

[0156] Therapeutic uses

[0157] disease

[0158] Preferably, the disease associated with or attributable to the expression of the PD-L1 protein according to the invention is cancer or an infectious disease.

[0159] More preferably, diseases related to or attributable to the PD-L1 protein are selected from:

[0160] - Unresectable metastatic melanoma, advanced metastatic non-small cell lung cancer (NSCLC), particularly advanced metastatic NSCLC, especially those that have progressed during or after platinum-based chemotherapy, advanced renal cell carcinoma, especially metastatic, locally advanced or metastatic urothelial carcinoma, especially those unresponsive to platinum-derived chemotherapy, prostate cancer, breast cancer, colorectal cancer, or any other cancer in which the PD-1 / PD-L1 axis is involved in the suppression of the antitumor immune response.

[0161] - Bacterial infections, such as pneumonia, meningitis, toxic shock syndrome, food poisoning, gastritis, ulcers, gonorrhea, boils, abscesses, impetigo, otitis media, pharyngitis, urinary tract infections and genital tract infections, and bronchopulmonary infections.

[0162] - Viral infections, such as influenza, measles, hepatitis B, hepatitis C, human immunodeficiency virus (HIV) infection, herpes-like virus infections, such as cytomegalovirus or Epstein-Barr virus, herpes, human papillomavirus (HPV) infection, and

[0163] - Fungal infections, such as blastomycosis, coccidioidomycosis, histoplasmosis, paracoccidioidomycosis, candidiasis, cryptococcosis, aspergillosis, mucormycosis, and Pneumocystis infection.

[0164] individual

[0165] The individual according to the invention is an animal, preferably a mammal or marsupial, more preferably a human, horse, cattle, pig, sheep, goat, camel, dromedary camel, dog or cat, and most preferably a human. According to the invention, preferably, the polypeptide of the invention is derived from the PD-L1 protein, which belongs to the same species as the individual using or administering the polypeptide.

[0166] application

[0167] Preferably, the polypeptides, pharmaceutical compositions, drugs, or products according to the invention are administered by means of oral, mucosal, especially sublingual, parenteral, intraperitoneal, percutaneous, intradermal, subcutaneous, intramuscular, intravenous, or intra-arterial administration.

[0168] dose

[0169] In the case of the present invention, the polypeptide according to the invention can be administered in a dosage range of, for example, 1 ng to 1 g, preferably 1 μg to 1 mg.

[0170] Pharmaceutically acceptable carriers

[0171] As understood in this article, "pharmaceutically acceptable carriers" include all compounds, particularly excipients, that can be applied to an individual in combination with a pharmacologically active ingredient.

[0172] adjuvant

[0173] Furthermore, particularly when used in vaccines or prophylactic settings, the polypeptide according to the invention can be combined with or conjugated with an adjuvant, or a pharmaceutical composition, drug, or product according to the invention can contain an adjuvant. The adjuvant can be of any type suitable for increasing the immune response to the polypeptide in an individual, animal, or human. Therefore, it can be a complete or incomplete adjuvant derived from Freund, Montanide ISA 51VG, aluminum hydroxide, or aluminum phosphate or calcium phosphate, with Montanide ISA 51VG and aluminum hydroxide or aluminum phosphate being preferred. The adjuvant can be combined with the polypeptide according to the invention by producing a 1 / 1 mixture of a volumetric adjuvant solution and a solution containing the polypeptide.

[0174] Other therapies

[0175] As understood herein, the expression “other therapies” means pharmacological or non-pharmacological therapies using at least one other compound different from the polypeptide according to the invention, such as radiotherapy, and in particular, anticancer therapy.

[0176] Other compounds

[0177] Other compounds that can be used to elicit an immune response against the PD-L1 protein according to the present invention may in particular be peptides derived from the PD-L1 protein that are different from the peptides of the present invention, or peptides derived from the PD-L protein.

[0178] In addition, other compounds intended to prevent or treat diseases, particularly cancer or infectious diseases, that are associated with or attributable to the expression of the PD-L1 protein or PD-1 protein can be anticancer chemotherapy compounds, anticancer immunotherapy compounds such as monoclonal antibodies, antibiotics, antiviral agents, especially interferon types, or antifungal agents.

[0179] Furthermore, particularly when used in a vaccine setting or contained in a vaccine or vaccine composition, the polypeptide according to the invention can be combined with other antigens designed to elicit an immune response against a target different from the PD-L1 protein (e.g., the PD-1 protein). This type of combination can be used to prepare multivalent vaccines.

[0180] As understood herein, the expression “combination” or “combination product” means that a polypeptide as defined above and another compound as defined above can be combined in the same pharmaceutical composition or the same drug and thus administered together or separately, i.e., according to different routes of administration and / or different administration regimens, provided that when they are administered separately, the prophylactic or therapeutic periods of the polypeptide as defined above and the other compound as defined above overlap entirely or partially.

[0181] Therefore, when the peptide and other compounds are administered separately, after the administration of the other compound as defined above, the peptide as defined above will preferably be administered within 24 hours, more preferably within 2 hours, and even more preferably within 1 hour, and its administration will optionally continue for the following days. Conversely, after the administration of the peptide as defined above, the other compound as defined above will preferably be administered within 24 hours, more preferably within 2 hours, and even more preferably within 1 hour, and its administration will optionally continue for the following days. In another preferred embodiment of the invention, when the peptide as defined above and the other compound as defined above are administered separately, they are administered substantially simultaneously.

[0182] The invention is further illustrated by the following non-limiting drawings and embodiments. Attached Figure Description

[0183] Figure 1

[0184] Figure 1 The relative amount of anti-hPD-L1 antibody present in serum (optical density measured by ELISA) represents the concentration of antibodies in the serum of SWISS mice immunized with peptides PPV-09-01, PPV-09-02, PPV-09-03, PPV-09-04, PPV-09-05, or PPV-09-06 diluted to 1 / 500 (n = 8 / group). The effective threshold is indicated at the 0.2 OD level.

[0185] Figure 2

[0186] Figure 2 This represents the percentage of PD1 / PD-L1 interaction neutralized by antibodies purified from rabbits (n=4 / group) immunized with peptides PPV-09-01, PPV-09-02, PPV-09-03, PPV-09-04, PPV-09-05, or PPV-09-06.

[0187] Example

[0188] Example 1: Recognition of intact human PD-L1 protein (hPD-L1) by serum from mice immunized with a peptide derived from hPD-L1.

[0189] Six peptides derived from the human PD-L1 protein were chemically synthesized and then cyclized by adding cysteine ​​residues to their ends to form disulfide bonds. These peptides were then coupled to the carrier protein CRM 197 (C-Reactive Material 197) using a GMBS coupling agent.

[0190] For each conjugate, specific pathogen-free SWISS mice (Janvier Labs, Le Genest-Saint-Isle, France) were immunized subcutaneously with 100 μg of peptide equivalents derived from human PD-L1 (PPV-09-01, PPV-09-02, PPV-09-03, PPV-09-04, PPV-09-05, or PPV-09-06; see Table 1), said peptide equivalents emulsified in adjuvant MontanideISA 51VG (n = 8 / conjugate). Mice received four subcutaneous injections at 15-day intervals (D0, D15, D30, and D45).

[0191] Table 1: Peptides derived from human PD-L1 used for immunity

[0192]

[0193] Sequence labeling of amino acid residues from human PD-L1 protein (Swissprot Q9NZQ7)

[0194] The relative amount of anti-PD-L1 antibody in the serum of D54 mice was assessed by ELISA (diluted to 1 / 500).

[0195] It can be observed that all tested conjugates produce antibodies that recognize the human PD-L1 protein. Figure 1 However, the immunogenicity of the conjugate PPV-09-03 was lower than that of the others.

[0196] Example 2: Neutralization of the biological activity of human PD-L1 by an antibody purified from rabbit serum, which was then immunized with a peptide derived from human PD-L1.

[0197] In a PD-L1 / PD-L1 interaction neutralizing cell assay, the neutralizing capacity of purified IgG from rabbit serum (n=4 / group) immunized with PPV-09-01, PPV-09-02, PPV-09-03, PPV-09-04, PPV-09-05, or PPV-09-06 peptides was evaluated (Promega, J1250). This assay is based on the interaction between two cell lines:

[0198] - Jurkat effector cell lines expressing the human PD-L1 gene and a luciferase gene controlled by the NFAT-RE response element.

[0199] -CHO-K1 cell line expressing the human PD-L1 gene and surface proteins for antigen-dependent activation of the TCR.

[0200] When the two lines are co-cultured, the interaction between the PD-L1 and PDL-1 proteins inhibits signal transduction via the TCR and the expression of the luciferase (no luminescence). Conversely, the addition of an anti-PD-L1 antibody to neutralize the PD-L1-PD-L1 interaction removes the inhibitory signal, leading to TCR activation and luminescence.

[0201] Experimental description:

[0202] Inoculation (D1): Inoculate 96-well flat-bottom plates, which have been treated for CHO-K1 cell culture. Incubate the plates at 37°C for 20 hours.

[0203] Sample incubation and display (D2): The antibody sample and Jurkat effector cells were distributed on a plate containing CHO-K1 cells. The plate was then incubated at 37°C for 6 hours.

[0204] Bio-Gio TM Add the reagent to each well. Incubate at room temperature for 30 minutes. Measure the luminescence using a luminometer.

[0205] It was observed that IgG from rabbits immunized with peptides PPV-09-01, PPV-09-02, PPV-09-03, PPV-09-04, PPV-09-05, or PPV-09-06 neutralized the PD-L1 protein-PD-L1 protein interaction to varying degrees, ranging from approximately 10% to 50%. Figure 2 ). sequence list <110> Peptinov Company National Institute of Technology <120> Anti-PD-L1 vaccine composition <130> B000082FR <160> 64 <170> PatentIn version 3.5 <210> 1 <211> 13 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 1 Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe 1 5 10 <210> 2 <211> 17 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 2 Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu Gln 1 5 10 15 Ile <210> 3 <211> 17 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 3 Val Tyr Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 Thr <210> 4 <211> 19 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 4 Asn Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu Leu 1 5 10 15 Thr Cys Gln <210> 5 <211> 16 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 5 Tyr Cys Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Glu 1 5 10 15 <210> 6 <211> 290 <212> PRT <213> Human <400> 6 Met Arg Ile Phe Ala Val Phe Ile Phe Met Thr Tyr Trp His Leu Leu 1 5 10 15 Asn Ala Phe Thr Val Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Met Thr Ile Glu Cys Lys Phe Pro Val Glu Lys Gln Leu 35 40 45 Asp Leu Ala Ala Leu Ile Val Tyr Trp Glu Met Glu Asp Lys Asn Ile 50 55 60 Ile Gln Phe Val His Gly Glu Glu Asp Leu Lys Val Gln His Ser Ser 65 70 75 80 Tyr Arg Gln Arg Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Val 115 120 125 Lys Val Asn Ala Pro Tyr Asn Lys Ile Asn Gln Arg Ile Leu Val Val 130 135 140 Asp Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr 145 150 155 160 Pro Lys Ala Glu Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser 165 170 175 Gly Lys Thr Thr Thr Thr Asn Ser Lys Arg Glu Glu Lys Leu Phe Asn 180 185 190 Val Thr Ser Thr Leu Arg Ile Asn Thr Thr Thr Asn Glu Ile Phe Tyr 195 200 205 Cys Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Glu Leu 210 215 220 Val Ile Pro Glu Leu Pro Leu Ala His Pro Pro Asn Glu Arg Thr His 225 230 235 240 Leu Val Ile Leu Gly Ala Ile Leu Leu Cys Leu Gly Val Ala Leu Thr 245 250 255 Phe Ile Phe Arg Leu Arg Lys Gly Arg Met Met Asp Val Lys Lys Cys 260 265 270 Gly Ile Gln Asp Thr Asn Ser Lys Lys Gln Ser Asp Thr His Leu Glu 275 280 285 Glu Thr 290 <210> 7 <211> 290 <212> PRT <213> Monkey <400> 7 Met Arg Ile Phe Ala Val Phe Ile Phe Thr Ile Tyr Trp His Leu Leu 1 5 10 15 Asn Ala Phe Thr Val Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Met Thr Ile Glu Cys Arg Phe Pro Val Glu Lys Gln Leu 35 40 45 Gly Leu Thr Ser Leu Ile Val Tyr Trp Glu Met Glu Asp Lys Asn Ile 50 55 60 Ile Gln Phe Val His Gly Glu Glu Asp Leu Lys Val Gln His Ser Asn 65 70 75 80 Tyr Arg Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn 85 90 95 Ala Ala Leu Arg Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Val 115 120 125 Lys Val Asn Ala Pro Tyr Asn Lys Ile Asn Gln Arg Ile Leu Val Val 130 135 140 Asp Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr 145 150 155 160 Pro Lys Ala Glu Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser 165 170 175 Gly Lys Thr Thr Thr Thr Asn Ser Lys Arg Glu Glu Lys Leu Leu Asn 180 185 190 Val Thr Ser Thr Leu Arg Ile Asn Thr Thr Ala Asn Glu Ile Phe Tyr 195 200 205 Cys Ile Phe Arg Arg Leu Gly Pro Glu Glu Asn His Thr Ala Glu Leu 210 215 220 Val Ile Pro Glu Leu Pro Leu Ala Leu Pro Pro Asn Glu Arg Thr His 225 230 235 240 Leu Val Ile Leu Gly Ala Ile Phe Leu Leu Leu Gly Val Ala Leu Thr 245 250 255 Phe Ile Phe Tyr Leu Arg Lys Gly Arg Met Met Asp Met Lys Lys Ser 260 265 270 Gly Ile Arg Val Thr Asn Ser Lys Lys Gln Arg Asp Thr Gln Leu Glu 275 280 285 Glu Thr 290 <210> 8 <211> 290 <212> PRT <213> Mouse <400> 8 Met Arg Ile Phe Ala Gly Ile Ile Phe Thr Ala Cys Cys His Leu Leu 1 5 10 15 Arg Ala Phe Thr Ile Thr Ala Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Met Glu Cys Arg Phe Pro Val Glu Arg Glu Leu 35 40 45 Asp Leu Leu Ala Leu Val Val Tyr Trp Glu Lys Glu Asp Glu Gln Val 50 55 60 Ile Gln Phe Val Ala Gly Glu Glu Asp Leu Lys Pro Gln His Ser Asn 65 70 75 80 Phe Arg Gly Arg Ala Ser Leu Pro Lys Asp Gln Leu Leu Lys Gly Asn 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Ile Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Val Asp 130 135 140 Pro Ala Thr Ser Glu His Glu Leu Ile Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Asn Ser Asp His Gln Pro Val Ser Gly 165 170 175 Lys Arg Ser Val Thr Thr Ser Arg Thr Glu Gly Met Leu Leu Asn Val 180 185 190 Thr Ser Ser Leu Arg Val Asn Ala Thr Ala Asn Asp Val Phe Tyr Cys 195 200 205 Thr Phe Trp Arg Ser Gln Pro Gly Gln Asn His Thr Ala Glu Leu Ile 210 215 220 Ile Pro Glu Leu Pro Ala Thr His Pro Pro Gln Asn Arg Thr His Trp 225 230 235 240 Val Leu Leu Gly Ser Ile Leu Leu Phe Leu Ile Val Val Ser Thr Val 245 250 255 Leu Leu Phe Leu Arg Lys Gln Val Arg Met Leu Asp Val Glu Lys Cys 260 265 270 Gly Val Glu Asp Thr Ser Ser Lys Asn Arg Asn Asp Thr Gln Phe Glu 275 280 285 Glu Thr 290 <210> 9 <211> 288 <212> PRT <213> Horse <400> 9 Met Arg Ile Val Ser Val Phe Thr Phe Met Ala Tyr Cys His Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Thr Lys Asp Leu Tyr Val Val Asp Tyr 20 25 30 Gly Ser Asn Val Thr Ile Glu Cys Lys Phe Pro Val Glu Glu Pro Leu 35 40 45 Asn Leu Ala Ala Leu Ile Val Tyr Trp Glu Met Glu Asn Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Glu Glu Asp Pro Lys Val Gln His Ser Ser 65 70 75 80 Tyr Ser Gln Arg Ala Arg Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Val Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Ser Ser Asp His Arg Val His Ser Gly 165 170 175 Lys Thr Thr Ile Thr Asn Ser Glu Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Arg Ile Asn Ala Thr Ala Asn Glu Ile Phe Tyr Cys 195 200 205 Thr Phe Arg Arg Ser Gly Leu Glu Glu Asn Ser Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Leu Ile Val Pro Ala Asn Lys Arg Thr His Leu Ala 225 230 235 240 Ile Leu Gly Val Ile Pro Leu Leu Leu Val Ala Leu Thr Ile Ile Ile 245 250 255 Cys Leu Lys Arg His Val Arg Met Met Asp Val Glu Lys Cys Ile Thr 260 265 270 Arg Asp Thr Asn Ser Lys Lys Gln Asn Asp Thr Gln Phe Glu Glu Thr 275 280 285 <210> 10 <211> 289 <212> PRT <213> Cattle <400> 10 Met Arg Ile Tyr Ser Val Leu Thr Phe Met Ala Tyr Cys Cys Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Ser Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Leu Glu Cys Arg Phe Pro Val Asp Lys Gln Leu 35 40 45 Asn Leu Leu Val Leu Val Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Lys Glu Asp Pro Asn Val Gln His Ser Ser 65 70 75 80 Tyr His Gly Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Tyr His Thr Ile Ser Val Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Asp Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser Gly 165 170 175 Lys Thr Ser Ile Thr Ser Ser Lys Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Arg Ile Asn Thr Thr Ala Asp Lys Ile Phe Tyr Cys 195 200 205 Thr Phe Arg Arg Leu Gly His Glu Glu Asn Asn Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Tyr Leu Asp Pro Ala Lys Lys Arg Asn His Leu Val 225 230 235 240 Thr Leu Gly Ala Leu Phe Leu Cys Leu Ser Val Thr Leu Ala Val Ile 245 250 255 Phe Cys Leu Lys Arg Asp Val Arg Met Met Asp Val Glu Lys Cys Asp 260 265 270 Thr Arg Asp Met Asn Ser Lys Gln Gln Asn Ala Thr Gln Phe Glu Glu 275 280 285 Thr <210> 11 <211> 287 <212> PRT <213> Pig <400> 11 Met Arg Ile Cys Ser Ile Phe Thr Phe Met Ala Tyr Cys Cys Leu Leu 1 5 10 15 Glu Ala Phe Thr Ile Thr Val Pro Lys Asp Met Tyr Glu Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Leu Glu Cys Arg Phe Pro Val Asp Lys Gln Leu 35 40 45 Asn Leu Leu Ala Leu Val Val Tyr Trp Glu Met Lys Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Glu Glu Asp Leu Asn Val Gln His Ser Ser 65 70 75 80 Tyr Ser Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ser Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Met Ser Leu Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Ser Ser Asp Tyr Gln Ile Leu Ser Gly 165 170 175 Lys Thr Thr Ile Thr Ser Ser Gln Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Arg Val Asn Ala Thr Thr Asn Glu Ile Phe Tyr Cys 195 200 205 Thr Phe Arg Arg Leu Gly Pro Glu Glu Asn Ser Thr Ala Val Leu Val 210 215 220 Ile Pro Glu Pro Tyr Val Asp Pro Ala Arg Lys Arg Thr His Trp Val 225 230 235 240 Ile Leu Gly Ala Leu Leu Leu Leu Ile Ala Val Thr Ala Ile Phe Cys 245 250 255 Leu Lys Arg Asn Val Arg Met Met Asp Val Glu Lys Cys Gly Ser Arg 260 265 270 Asp Met Lys Ser Glu Lys Gln Asn Asp Thr Gln Phe Glu Glu Thr 275 280 285 <210> 12 <211> 316 <212> PRT <213> Sheep <400> 12 Met Arg Ile Tyr Ser Val Leu Thr Phe Met Ala Tyr Cys Cys Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Leu Glu Cys Arg Phe Pro Val Asp Gln Gln Leu It should be noted that there seems to be a misspelling in "绵羊" which should be "绵羊(sheep)" in Chinese. And in the translation, I've corrected it to "Sheep" for better understanding. Also, the "35" in the original text should be "35" which I've corrected in the translation for consistency.35 40 45 Asn Leu Leu Val Leu Val Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Lys Glu Asp Leu Asn Val Gln His Ser Ser 65 70 75 80 Tyr His Gly Arg Ala Gln Leu Leu Lys Asp Gln Leu Ser Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Tyr Pro Thr Val Ser Val Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Asp Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser Gly 165 170 175 Lys Thr Ser Ile Thr His Ser Lys Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Arg Ile Asn Thr Thr Ala Asp Lys Ile Phe Tyr Cys 195 200 205 Thr Phe Arg Arg Leu Gly His Glu Glu Asn Asn Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Tyr Pro Asp Pro Ala Lys Thr Arg Asn His Leu Val 225 230 235 240 Ile Leu Gly Ala Leu Phe Leu Phe Leu His Val Thr Leu Ala Val Ile 245 250 255 Phe Cys Leu Lys Arg Asn Val Arg Lys Met Asp Val Glu Lys Cys Gly 260 265 270 Thr Gln Asp Met Asn Ser Lys Gln Gln Asn Gly Lys Asn Phe Ser Arg 275 280 285 Asp Trp Lys Leu Lys Lys Gly Asn Lys Lys Leu Lys Asn Lys Gly Lys 290 295 300 Ala Ile Ile Ile Ile Ser Pro Tyr Phe Thr Glu Cys 305 310 315 <210> 13 <211> 289 <212> PRT <213> Goat <400> 13 Met Arg Ile Tyr Ser Val Leu Thr Phe Met Ala Tyr Cys Cys Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Leu Glu Cys Arg Phe Pro Val Asp Gln Gln Leu 35 40 45 Asn Leu Leu Val Leu Val Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Lys Glu Asp Leu Asn Val Gln His Ser Ser 65 70 75 80 Tyr His Gly Arg Ala Gln Leu Leu Lys Asp Gln Leu Ser Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Tyr His Thr Ile Ser Val Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Asp Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser Gly 165 170 175 Lys Thr Ser Ile Thr Asn Ser Lys Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Arg Ile Asn Thr Thr Ala Asp Lys Ile Phe Tyr Cys 195 200 205 Thr Phe Arg Arg Leu Gly His Glu Glu Asn Asn Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Tyr Pro Asp Pro Ala Lys Thr Arg Asn His Leu Val 225 230 235 240 Ile Leu Gly Ala Leu Phe Leu Phe Leu His Val Thr Leu Ala Val Ile 245 250 255 Phe Cys Leu Lys Arg Asn Val Arg Lys Met Asp Val Glu Lys Cys Gly 260 265 270 [[ID=2,0]] Thr Gln Asp Met Asn Ser Lys Gln Gln Asn Ala Thr His Phe Glu Glu 275 280 285 Thr <210> 14 <211> 289 <212> PRT <213> Bactrian camel[[ID=,34]] <400> 14 Met Arg Ile Cys Ser Val Phe Thr Phe Val Ala Tyr Cys Cys Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Met Glu Cys Lys Phe Pro Val Asp Lys Gln Leu 35 40 45 Asn Leu Leu Ala Leu Val Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Glu Glu Asp Leu Asn Val Gln His Ser Ser 65 70 75 80 Tyr Ser Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Met Asp 130 135 140 Pro Val Thr Ala Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Ser Ser Asp His Arg Val Leu Ser Gly 165 170 175 Lys Thr Thr Val Thr Ser Ser Gln Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Ser Leu Arg Ile Asn Thr Thr Ala Asn Glu Ile Phe Tyr Cys 195 200 205 Ile Phe Arg Arg Leu Gly His Glu Glu Asn Ser Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Tyr Val Asp Pro Ala Lys Lys Arg Thr His Leu Val 225 230 235 240 Val Leu Gly Ala Ala Leu Leu Val Leu Gly Val Ile Leu Thr Val Ile 245 250 255 Phe Cys Leu Lys Arg Asp Val Arg Met Met Asp Val Glu Lys Cys Val 260 265 270 Thr Arg Asp Thr Asn Ser Arg Lys Gln Asn Asp Pro Gln Phe Glu Glu 275 280 285 Thr <210> 15 <211> 289 <212> PRT <213> Bactrian camel <400> 15 Met Arg Ile Cys Ser Val Phe Thr Phe Val Ala Tyr Cys Cys Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Met Glu Cys Lys Phe Pro Val Asp Lys Gln Leu 35 40 45 Asn Leu Leu Ala Leu Val Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Glu Glu Asp Leu Asn Val Gln His Ser Ser 65 70 75 80 Tyr Ser Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Met Asp 130 135 140 Pro Val Thr Ala Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Ser Ser Asp His Arg Val Leu Ser Gly 165 170 175 Lys Thr Thr Val Thr Ser Ser Gln Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Ser Leu Arg Ile Asn Thr Thr Ala Asn Glu Ile Phe Tyr Cys 195 200 205 Ile Phe Arg Arg Leu Gly His Glu Glu Asn Ser Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Tyr Val Asp Pro Ala Lys Lys Arg Thr His Leu Val 225 230 235 240 Val Leu Gly Ala Ala Leu Leu Val Leu Gly Val Ile Leu Thr Val Ile 245 250 255 Phe Cys Leu Lys Arg Asp Val Arg Met Met Asp Val Glu Lys Cys Val 260 265 270 Thr Arg Asp Thr Asn Ser Arg Lys Gln Asn Asp Pro Gln Phe Glu Glu 275 280 285 Thr <210> 16 <211> 289 <212> PRT <213> Dromedary <400> 16 Met Arg Ile Cys Ser Val Phe Thr Phe Val Ala Tyr Cys Cys Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Ser Asn Val Thr Met Glu Cys Lys Phe Pro Val Asp Lys Gln Leu 35 40 45 Asn Leu Leu Ala Leu Val Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Glu Glu Asp Leu Asn Val Gln His Ser Ser 65 70 75 80 Tyr Ser Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Met Asp 130 135 140 Pro Val Thr Ala Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Ser Ser Asp His Arg Val Leu Ser Gly 165 170 175 Lys Thr Thr Val Thr Ser Ser Gln Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Ser Leu Arg Ile Asn Thr Thr Ala Asn Glu Ile Phe Tyr Cys 195 200 205 Ile Phe Arg Arg Leu Gly His Glu Glu Asn Ser Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Pro Tyr Val Asp Pro Ala Lys Lys Arg Thr His Leu Val 225 230 235 240 Val Leu Gly Ala Ala Leu Leu Val Leu Gly Ile Ile Leu Thr Val Ile 245 250 255 Phe Cys Leu Lys Arg Asp Val Arg Met Met Asp Val Glu Lys Cys Val 260 265 270 Thr Arg Asp Thr Asn Ser Arg Lys Gln Asn Asp Pro Gln Phe Glu Glu 275 280 285 Thr <210> 17 <211> 289 <212> PRT <213> dog <400> 17 Met Arg Met Phe Ser Val Phe Thr Phe Met Ala Tyr Cys His Leu Leu 1 5 10 15 Lys Ala Phe Thr Ile Thr Val Ser Lys Asp Leu Tyr Val Val Glu Tyr 20 25 30 Gly Gly Asn Val Thr Met Glu Cys Lys Phe Pro Val Glu Lys Gln Leu 35 40 45 Asn Leu Phe Ala Leu Ile Val Tyr Trp Glu Met Glu Asp Lys Lys Ile 50 55 60 Ile Gln Phe Val Asn Gly Lys Glu Asp Leu Lys Val Gln His Ser Ser 65 70 75 80 Tyr Ser Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asp Val Arg Leu Gln Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Gly Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val His Ala Pro Tyr Arg Asn Ile Ser Gln Arg Ile Ser Val Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Met Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Glu Ala Glu Val Ile Trp Thr Ser Ser Asp His Arg Val Leu Ser Gly 165 170 175 Lys Thr Thr Ile Thr Asn Ser Asn Arg Glu Glu Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Asn Ile Asn Ala Thr Ala Asn Glu Ile Phe Tyr Cys 195 200 205 Thr Phe Gln Arg Ser Gly Pro Glu Glu Asn Asn Thr Ala Glu Leu Val 210 215 220 Ile Pro Glu Arg Leu Pro Val Pro Ala Ser Glu Arg Thr His Phe Met 225 230 235 240 Ile Leu Gly Pro Phe Leu Leu Leu Leu Gly Val Val Leu Ala Val Thr 245 250 255 Phe Cys Leu Lys Lys His Gly Arg Met Met Asp Val Glu Lys Cys Cys 260 265 270 Thr Arg Asp Arg Asn Ser Lys Lys Arg Asn Asp Ile Gln Phe Glu Glu 275 280 285 Thr <210> 18 <211> 291 <212> PRT <213> Cat <400> 18 ​​​​​​​​​​​​​​​​​​65 70 75 80 Tyr Ser Gln Arg Ala Gln Leu Leu Lys Asp Gln Leu Phe Leu Gly Lys 85 90 95 Ala Ala Leu Gln Ile Thr Asn Val Thr Leu Glu Asp Ala Gly Val Tyr 100 105 110 Cys Cys Leu Ile Gly Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu 115 120 125 Lys Val His Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Val Asp 130 135 140 Pro Val Thr Ser Glu His Glu Leu Met Cys Gln Ala Glu Gly Tyr Pro 145 150 155 160 Thr Ala Glu Val Ile Trp Thr Asn Ser Ala His Gln Val Leu Asn Gly 165 170 175 Lys Thr Ile Ile Ser Val Ser Asn Met Glu Thr Lys Leu Phe Asn Val 180 185 190 Thr Ser Thr Leu Arg Ile Asn Thr Thr Ala Asn Glu Ile Phe Tyr Cys 195 200 205 Thr Phe Leu Gln Arg Ser Ser Pro Glu Gly Asn Ser Thr Ala Glu Leu 210 215 220 Val Ile Pro Glu Pro Phe Leu Val Pro Ala Asn Glu Arg Thr His Phe 225 230 235 240 Met Ile Leu Gly Ala Ile Leu Leu Phe Leu Val Val Val Pro Ala Val 245 250 255 Thr Phe Cys Leu Lys Lys Arg Asp Val Arg Thr Met Asp Val Glu Lys 260 265 270 Cys Asp Thr Ala Asp Met Asn Ser Lys Lys Gln Asn Asp Leu Gln Phe 275 280 285 Glu Glu Thr 290 <210> 19 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Variants of PD-L1 fragments <400> 19 Val Tyr Arg Ser Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 Thr <210> 20 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Variants of PD-L1 fragments <400> 20 Asn Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu Leu 1 5 10 15 Thr Ser Gln <210> twenty one <211> 16 <212> PRT <213> Artificial sequence <220> <223> Variants of PD-L1 fragments <400> twenty one Tyr Ser Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Glu 1 5 10 15 <210> twenty two <211> 14 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> twenty two Cys Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe 1 5 10 <210> twenty three <211> 14 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> twenty three Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe Cys 1 5 10 <210> twenty four <211> 18 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> twenty four Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu Gln 1 5 10 15 Ile Cys <210> 25 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 25 Cys Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu 1 5 10 15 Gln Ile <210> 26 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 26 Cys Val Tyr Arg Ser Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg 1 5 10 15 Ile Thr <210> 27 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 27 Val Tyr Arg Ser Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 Thr Cys <210> 28 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 28 Cys Asn Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu 1 5 10 15 Leu Thr Ser Gln 20 <210> 29 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 29 Asn Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu Leu 1 5 10 15 Thr Ser Gln Cys 20 <210> 30 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 30 Cys Tyr Ser Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala 1 5 10 15 Glu <210> 31 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 31 Tyr Ser Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Glu 1 5 10 15 Cys <210> 32 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 32 Cys Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe 1 5 10 <210> 33 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 33 Cys Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu Gln 1 5 10 15 Ile <210> 34 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 34 Cys Tyr Arg Ser Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 Thr <210> 35 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 35 Cys Gln Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe Lys 1 5 10 15 <210> 36 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 36 Gln Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe Lys Cys 1 5 10 15 <210> 37 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 37 Cys Gln Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala 1 5 10 15 Leu Gln Ile Lys 20 <210> 38 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 38 Gln Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu 1 5 10 15 Gln Ile Lys Cys 20 <210> 39 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 39 Cys Gln Val Tyr Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys 1 5 10 15 Arg Ile Thr Lys 20 <210> 40 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 40 Gln Val Tyr Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg 1 5 10 15 Ile Thr Lys Cys 20 <210> 41 <211> twenty one <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 41 Cys Lys Asn Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His 1 5 10 15 Glu Leu Thr Cys Gln 20 <210> 42 <211> twenty one <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 42 Lys Asn Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu 1 5 10 15 Leu Thr Cys Gln Cys 20 <210> 43 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 43 Cys Lys Tyr Ser Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr 1 5 10 15 Ala Glu Gln <210> 44 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <400> 44 Lys Tyr Ser Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala 1 5 10 15 Glu Gln Cys <210> 45 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <220> <221> MISC_FEATURE <222> (15)..(15) <223> Leucine (Nle) <400> 45 Cys Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Cys Gly Xaa Glu 1 5 10 15 Cys <210> 46 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <220> <221> MISC_FEATURE <222> (21)..(21) <223> N-leucine (Nle) <400> 46 Cys Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu Gln 1 5 10 15 Ile Cys Gly Glu Xaa Glu Cys 20 <210> 47 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <220> <221> MISC_FEATURE <222> (14)..(14) <223> N-leucine (Nle) <400> 47 Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Cys Arg Xaa Arg Cys 1 5 10 15 <210> 48 <211> twenty one <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <220> <221> MISC_FEATURE <222> (19)..(19) <223> N-leucine (Nle) <400> 48 Cys Tyr Arg Ser Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 Cys Gly Xaa Arg Cys 20 <210> 49 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <220> <221> MISC_FEATURE <222> (21)..(21) <223> N-leucine (Nle) <400> 49 Cys Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu Leu 1 5 10 15 Thr Cys Gly Glu Xaa Glu Cys 20 <210> 50 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Immunogenic peptides <220> <221> MISC_FEATURE <222> (17)..(17) <223> N-leucine (Nle) <400> 50 Cys Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Cys Gly 1 5 10 15 Xaa Ser Cys <210> 51 <211> 12 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 51 Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln Phe 1 5 10 <210> 52 <211> 12 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 52 Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln 1 5 10 <210> 53 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 53 Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln 1 5 10 <210> 54 <211> 16 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 54 Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu Gln Ile 1 5 10 15 <210> 55 <211> 16 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 55 Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala Leu Gln 1 5 10 15 <210> 56 <211> 16 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 56 Tyr Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr 1 5 10 15 <210> 57 <211> 16 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 57 Val Tyr Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 <210> 58 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 58 Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr 1 5 10 <210> 59 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 59 Tyr Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 <210> 60 <211> 17 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 60 Gln Arg Ile Leu Val Val Asp Pro Val Thr Ser Glu His Glu Leu Thr 1 5 10 15 Cys <210> 61 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 61 Tyr Cys Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala 1 5 10 15 <210> 62 <211> 14 <212> PRT <213> Artificial sequence <220> <223> PD-L1 fragment <400> 62 Cys Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala 1 5 10 <210> 63 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Variants of PD-L1 fragments <400> 63 Tyr Arg Ser Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile 1 5 10 15 <210> 64 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Variants of PD-L1 fragments <400> 64 Tyr Ser Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala 1 5 10 15

Claims

1. A polypeptide, which is composed of the following: - The first sequence, which consists of SEQ ID NO:45; or - The second sequence, which consists of SEQ ID NO:46; or - A third sequence, consisting of SEQ ID NO:47 or 48; or - The fourth sequence, which consists of SEQ ID NO:49; or - The fifth sequence, which consists of SEQ ID NO:50; The polypeptide described therein is in a cyclized form.

2. The polypeptide according to claim 1, wherein the polypeptide is linked to a carrier molecule.

3. A nucleic acid encoding a polypeptide as defined in claim 1 or 2.

4. A pharmaceutical composition comprising the following active substances: - at least one polypeptide as defined in claim 1 or 2, or - At least one nucleic acid as defined in claim 3, Optionally combined with at least one pharmaceutically acceptable carrier.

Images