Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
  • C07K16/2827—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against B7 molecules, e.g. CD80, CD86
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
  • C07K14/70503—Immunoglobulin superfamily
  • C07K14/70532—B7 molecules, e.g. CD80, CD86
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/30—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies

Definitions

• the present invention provides polypeptides useful for eliciting an immune response directed against the PD-L1 protein.
• the two most important are its targeting by monoclonal antibodies specific for cytotoxic T-cell-associated protein 4 (CTLA-4) and interaction between the programmed cell death protein 1 (PD-1) and the ligand of this programmed cell death protein 1 (PD-L1).
• CTLA-4 cytotoxic T-cell-associated protein 4
• PD-1 programmed cell death protein 1
• PD-L1 programmed cell death protein 1
• the inhibition of PD-L1 signaling has been proposed as a means of improving T cell immunity for the treatment of cancer (anfi-fumoral immunity) but also in the treatment of infections (acute persistent infection and chronic).
• FDA US Federal Drug Administration
• pembrolizumab an anti-PD-1 monoclonal antibody (mAb) approved for people with unresectable metastatic melanoma or advanced non-small-cell metastatic lung carcinoma (NSCLC) whose tumors express PD-1 L1;
• nivolumab an anti-PD-1 monoclonal antibody (mAb) approved for unresectable or metastatic melanoma, advanced metastatic NSCLC progressing with or after platinum-based chemotherapy, and advanced renal cell carcinoma, including metastatic ; e ⁇
• Atezolizumab a recently approved anti-PD-Ll monoclonal antibody (mAb) for the treatment of locally advanced or metastatic urothelial carcinoma that does not respond to platinum derivative chemotherapy; e ⁇
• Avelumab a recently approved anti-PD-Ll monoclonal antibody (mAb) for the treatment of metastatic Merkel cell carcinoma.
• PD-L1 targeting ICBs have been shown to be highly effective in melanoma, NSCLC and renal cell carcinoma.
• the present invention results from the unexpected finding by the inventors that polypeptides derived from sequences extending residues of amino acids 55 to 67, 85 to 101, 11 to 127, 138 to 156, e 208 to 223 of the human PD-L1 protein allowed the production of antibodies neutralizing the PD-L1 protein by mice to which they were administered.
• the present invention thus relates to a polypeptide comprising or consisting of:
• a first sequence consisting of at least 8 contiguous amino acid residues selected from the sequence extending from amino acid residues 55 to 67 of the PD-L1 protein and of at most 30 acid residues contiguous amines 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
• a second sequence consisting of at least 8 contiguous amino acid residues selected from the sequence extending from amino acid residues 85 to 101 of the PD-L1 protein and of at most 30 acid residues contiguous amines 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
• a third sequence consisting of at least 8 contiguous amino acid residues selected from the sequence extending from amino acid residues 11 to 127 of the PD-L1 protein and at most 30 residues of contiguous amino acids 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
• a fourth sequence consisting of at least 8 contiguous amino acid residues selected from the sequence extending from amino acid residues 138 to 156 of the PD-L1 protein and of at most 30 acid residues contiguous amines 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
• a fifth sequence consisting of at least 8 contiguous amino acid residues selected from the sequence extending from amino acid residues 208 to 223 of the PD-L1 protein and of at most 30 acid residues contiguous amines selected from the complete sequence of the PD-L1 protein, or a variant sequence having at least 75% identity with the fifth sequence;
• polypeptide is different from the PD-L1 protein and does not consist of a portion of more than 30 contiguous amino acid residues of the PD-L1 protein, and
• polypeptides respectively consisting of variant sequences of the first, second, third, fourth, and fifth sequences elicit an immune response directed against the PD-L1 protein.
• the invention relates more particularly to a polypeptide comprising or consisting of:
• 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
• 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
• a fourth sequence consisting of SEQ ID NO: 4 or 60, or a variant sequence exhibiting at least 75% identity with the fourth sequence; and or
• a fifth sequence consisting of SEQ ID NO: 5, 61 or 62, or a variant sequence exhibiting at least 75% identity with the fifth sequence;
• polypeptides consisting respectively of variant sequences of the first, second, third, fourth and fifth sequences elicit an immune response directed against the PD-L1 protein.
• the present invention also relates to a nucleic acid encoding a polypeptide as defined above, or the complement thereof.
• the present invention also relates to:
• the medicament, in particular the vaccine, as defined above also comprises at least one other compound intended for the prevention or treatment of a disease linked or due to the expression of PD-L1 protein or PD-1 protein, cancer or infectious disease.
• the present invention also relates to a pharmaceutical composition, in particular a vaccine composition, comprising, with the filler of active substance:
• the pharmaceutical composition, in particular vaccine, as defined above also comprises at least one other compound intended for the prevention or treatment of a disease linked or due to the expression of PD-L1 protein or PD-1 protein, cancer or infectious disease.
• the present invention also relates to the use of a polypeptide as defined above, for the preparation of an antibody, an antibody fragment or an apfamer.
• the present invention also relates to a method for preparing an antibody, an antibody fragment or an apfamer comprising a step of administering a polypeptide as defined above to an antibody-producing organism or an affinity screening step of an antibody, an antibody fragment or an apfamer which binds to the polypeptide as defined above.
• the present invention also relates to an antibody, an antibody fragment, or an anti-PD-L1 apfamer specifically directed against the polypeptide as defined above, provided that the polypeptide does not comprise more than two amino acid residues. in addition to the first, second, third, fourth, or fifth sequences or their respective variant sequences.
• the present invention also relates to an antibody, an antibody fragment, or an apfamer as defined above for use with a fife of medicine.
• the medicament as defined above also comprises at least one other compound intended for the prevention or treatment of a disease linked or due to the expression of the PD-L1 protein or of the PD-1 protein, of a cancer or an infectious disease.
• the present invention also relates to a pharmaceutical composition
• a pharmaceutical composition comprising, with the fuid of active principle, an antibody, an antibody fragment, or an apfamer as defined above, optionally in combination with a pharmaceutically acceptable vehicle.
• the pharmaceutical composition as defined above also comprises at least one other compound intended for the prevention or treatment of a disease linked or due to the expression of the PD-protein. L1 or PD-1 protein, cancer or infectious disease.
• the present invention also relates to a polypeptide as defined above, a nucleic acid as defined above, or a pharmaceutical composition as defined above, for use in a method for eliciferating an immune response against the protein PD-L1 in an individual.
• the polypeptide, nucleic acid or pharmaceutical composition is used in combination with at least one other compound useful for eliciting an immune response directed against the PD-1 protein.
• the present invention also relates to a method for eliciting an immune response directed against the PD-L1 protein in an individual, comprising administering to the individual an effective amount of a polypeptide as defined above, a nucleic acid as defined above, or a pharmaceutical composition as defined above.
• the polypeptide, nucleic acid or pharmaceutical composition is administered in combination with at least one other compound useful for eliciting an immune response directed against the PD-L1 protein.
• the present invention also relates to the use of a polypeptide as defined above or a nucleic acid as defined above for the preparation of a medicament for eliciting an immune response directed against the PD-1 protein in an individual.
• the medicament also comprises at least one other compound useful for eliciting an immune response directed against the PD-L1 protein.
• the present invention also relates to a polypeptide as defined above, a nucleic acid as defined above, a pharmaceutical composition as defined above, or an antibody, an antibody fragment or an apfamer as defined herein. above, for use in a method of preventing or treating a disease related to or due to expression of PD-1 protein or PD-L1 protein in an individual.
• the polypeptide, nucleic acid, pharmaceutical composition, or antibody, antibody fragment or aptamer is used in combination with at least one other therapy for prevention or treatment of a disease related to the expression of PD-1 protein or PD-L1 protein, cancer or infectious disease.
• the present invention also relates to a method of preventing or treating a PD-L1-related or PD-related disease in an individual, comprising administering to the individual an effective amount of a polypeptide as defined herein. above, a nucleic acid as defined above, a pharmaceutical composition as defined above, or an antibody, an antibody fragment or an aptamer as defined hereinabove. above.
• the method comprises at least one other therapy for the prevention or treatment of a disease related to the expression of the PD-1 protein or of the PD-L1 protein, d 'a cancer or an infectious disease.
• the present invention also relates to the use of a polypeptide as defined above, a nucleic acid as defined above, or an antibody, an antibody fragment or an aptamer such as as defined above, for the preparation of a medicament for the prevention or treatment of a PD-L1-related or PD-related disease in an individual.
• the medicament comprises at least one other compound intended for the prevention of a disease linked or due to the expression of the PD-1 protein or of the PD-L1 protein, of cancer or an infectious disease.
• the present invention also relates to products containing:
• the term “comprising” means “including”, “containing” or “encompassing”, that is to say that when an object “includes” one or more elements, other elements that those mentioned can also be included in the object.
• the expression “consisting of” means “consisting of”, that is, when an object “consists of” one or more elements, the object can not include other elements than those mentioned.
• the PD-L1 protein, the protein ligand of the PD-1 protein, also called the differentiation cluster 274 (cluster of differentiation 274, CD274), is well known to those skilled in the art.
• the PD-L1 protein according to the invention is selected from the group consisting of human PD-L1 protein, mouse PD-L1 protein, monkey PD-L1 protein, PD-L1 protein, Horse L1, bovine PD-L1 protein, pig PD-L1 protein, sheep PD-L1 protein, goat PD-L1 protein, camel PD-L1 protein, camel PD-L1 protein, PD-L1 dog protein, e ⁇ of the PD-L1 cat protein.
• the PD-L1 protein is the human PD-L1 protein.
• the monkey PD-L1 protein as described in the Genbank database under the reference NPJD01077358.1 ⁇ consisting of SEQ ID NO: 7
• the mouse PD-L1 protein (mPD-1) is as described in the UniProt / Swissprot database under the reference Q9EP73 e ⁇ es consisting of SEQ ID NO: 8,
• bovine PD-L1 protein as described in the Genbank database under the reference NP_001 156884.1 and consisting of SEQ ID NO: 10,
• camel PD-L1 protein as described in the Genbank database under the reference XP_01441 6021 .1 or XP_010958932.1 and es ⁇ consisting of SEQ ID NO: 14 or 15,
• the dog PD-L1 is as described in the UniProf / SwissProf database under the reference E2RKZ5 and consists of SEQ ID NO: 17,
• chat PD-L1 as described in the Genbank database under the reference XPJD06939101 .1 and es ⁇ consisting of SEQ ID NO: 18.
• the amino acid residue numbering of the PD-L1 protein begins on the first amino acid residue, typically a methionine (M), forming the N-terminus of complete PD-L1. encoded by the open reading frame of the PD-L1 gene, that is, including its signal peptide.
• M methionine
• the amino acid residue numbering of the PD-L1 protein used herein is defined by reference to the human PD-L1 protein.
• the first sequence, the second sequence, the third sequence, the fourth sequence and the fifth sequence consist of at least 8, 9, 10, 1 1, 12 contiguous amino acid residues respectively chosen within of the sequence extending from amino acid residues 55 to 67, 85 to 1 01, 1 1 1 to 1 27, 138 to 1 56, e 208 to 223 of the PD-L1 protein or are constituted respectively at least of the sequence extending from amino acid residues 55 to 67, 85 to 101, 11 to 1 27, 138 to 156, e 208 to 223 of the PD-L1 protein.
• the first sequence, the second sequence, the third sequence, the fourth sequence and the fifth sequence according to the invention are respectively at most 29, 28, 27, 26, 25, 24, 23, 22 , 21, 20 contiguous amino acid residues selected from the complete sequence of the PD-L1 protein, or are respectively at most amino acid residues 55 to 67, 85 to 1 01, 1 1 1 to 127, 1 38 to 1 56, e 208 to 223 of the PD-L1 protein.
• the first sequence according to the invention consists of SEQ ID NO: 1, 51, 52 or 53,
• the second sequence according to the invention consists of SEQ ID NO: 2, 54 or 55,
• the third sequence according to the invention consists of SEQ ID NO: 3, 56, 57, 58 or 59,
• the fourth sequence according to the invention consists of SEQ ID NO: 4 or
• the fifth sequence according to the invention consists of SEQ ID NO: 5, 61 or 62.
• a variant sequence according to the invention which has at least 75% identity with one of the first, second, third, fourth, and fifth sequences above, preferably has at least 80%, 85%, 90% or more. %, 95%, or 98% identity with one of the first, second, third, fourth, and fifth sequences above.
• the percentage identity between two peptide sequences can be determined by performing optimal alignment over the entire length of the sequences, by determining the number of aligned positions for which the amino acids are identical in each sequence and by dividing this number by the total number of amino acids in the longest of the two sequences.
• the optimal alignment is the one that gives the highest percentage of identity between the two sequences.
• a 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.
• the variant sequence according to the invention is such that a polypeptide consisting of the variant sequence must make it possible to elicit an immune response directed against the PD-L1 protein; that is to say that the administration of such a peptide, possibly cyclized by formation of at least one infer-cysteine disulfide bridge, if necessary after adding one or two cysteines within the peptide, and / or at its N-ferminal end and / or at its C-terminal end, the peptide optionally being bound to a carrier molecule, in particular a carrier protein, such as KLH (Keyhole Limpet Hemocyanin), to an animal, such as a mouse , a rat or a rabbit, causes the production of antibodies directed against a PD-L1, in particular a PD-L1 of the same species as that to which belongs the sequence with which the variant sequence has the highest percentage of identity .
• a carrier molecule in particular a carrier protein, such as KLH (Keyhole Limpet Hemocyan
• the antibodies elicited by administration of the peptide are blocking or neutralizing, that is to say, they prevent the PD-L1 protein from exercising all or part, especially at least 10%, 25%, 50%. %, 75%, of its activity, for example measured in vitro.
• the activity of PD-L1 is preferably a binding to the PD-1 protein, which can be measured as in Example 2 which follows.
• a variant sequence according to the invention is selected from the group consisting of:
• VYRSMISYGGADYKRIT (SEQ ID NO: 19) derives from VYRCMISYGGADYKRIT (SEQ ID NO: 3) by the substitution of cysteine (C) in the fourth position by a serine (S).
• YRSMISYGGADYKRI (SEQ ID NO: 63) derives from YRCMISYGGADYKRI (SEQ ID NO: 59) by the substitution of cysteine (C) in third position by a serine (S).
• NQRILVVDPVTSEHELTSQ (SEQ ID NO: 20) derives from NQRILVVDPVTSEHELTCQ (SEQ ID NO: 4) by the substitution of cysteine (C) in penultimate position by a serine (S).
• YSTFRRLDPEENHTAE (SEQ ID NO: 21) is derived from YCTFRRLDPEENHTAE (SEQ ID NO: 5) by the substitution of cysteine (C) in the second position by a serine (S).
• YSTFRRLDPEENHTA (SEQ ID NO: 64) derives from YCTFRRLDPEENHTA (SEQ ID NO: 61) by the substitution of cysteine (C) in the second position by a serine (S).
• the polypeptide according to the invention preferably comprises at most 200, 150, 100, 90, 80, 70, 60, 50, 40 or 30 amino acid residues. It is different from the PD-L1 protein and does not consist of a portion of more than 30 contiguous amino acid residues of the PD-L1 protein. As is well understood by those skilled in the art, this does not exclude that it may consist of two or more portions of the PD-L1 protein of not more than 30 contiguous amino acid residues, since these portions are not arranged to reconstitute a portion of the PD-L1 protein of more than 30 contiguous amino acids.
• polypeptide according to the invention may include several repetitions, for example 2, 3, 4, 5, 10 or 20 repetitions, respectively of the first, second, third, fourth and fifth sequences e ⁇ of the variant sequence according to the invention.
• polypeptide according to the invention may also comprise additional sequences not originating from the PD-L1 protein.
• additional sequences may in particular provide physicochemical characteristics allowing improved structural presentation or improved solubility of the polypeptide according to the invention relative to a similar polypeptide but which will not include these additional sequences.
• the additional sequences may also comprise one or more peptide linker sequences, that is to say peptide linker, which are useful for an particularly binding to a carrier molecule.
• peptide linker sequences typically comprise from 1 to 10, especially from 4 to 6, amino acid residues.
• sequences not coming from the PD-L1 protein may also include epitopes belonging to other proteins, for eliciting or generating an immune response directed against these other proteins.
• polypeptide according to the invention may comprise exogenous (s) T epitope sequences, preferably universal, which makes it possible to enhance the immunogenicity of the polypeptide according to the invention.
• the polypeptide according to the invention may also comprise at least one sequence of a carrier protein, for example a virus-like particle (VLP), as described in particular in International Application WO 05/117983 for TNF.
• a carrier protein for example a virus-like particle (VLP), as described in particular in International Application WO 05/117983 for TNF.
• VLP virus-like particle
• the polypeptide according to the invention may be in linear form or in cyclized form.
• the polypeptide according to the invention is in cyclized form. This cyclization may be of any type known to those skilled in the art.
• the choice of the cyclization strategy according to the invention can in particular take into account the best antigenic presentation of the epitopes contained in the polypeptide according to the invention, and relate only to a part of the polypeptide (cyclization within the sequence). .
• the polypeptide of the invention when the polypeptide of the invention is in a cyclized form, only a portion of the polypeptide may be included in one cycle while the remainder of the polypeptide is in linear form.
• this cyclization can be carried out in several different ways, for example: from its N-terminal end C-terminus, from its N-terminus to a side chain, a side chain at its C-terminal end or between two side chains.
• lactamization lactonization or the formation of a disulfide bridge.
• cysteines may already be present in the variant sequence according to the invention or in the first, second, third, fourth and fifth sequences according to the invention, or else to be added within these sequences, as well as at their N-terminus and / or C-terminus.
• polypeptide according to the invention may comprise post-translational modifications, such as glycosylations, methylations, acylations, in particular by fatty acids, or phosphorylations.
• post-translational modifications such as glycosylations, methylations, acylations, in particular by fatty acids, or phosphorylations.
• the N-terminus of the polypeptide according to the invention may be acetylated at the C-terminus may be modified by amidation.
• polypeptide according to the invention may also comprise one or more analogs or amino acid derivatives, including non-natural or non-standard amino acids, in particular norleucine (Nie).
• Nie norleucine
• polypeptide according to the invention is fixed or bound, in particular by covalent bonding, to a carrier molecule, in particular a carrier protein.
• the carrier molecule may be Keyhole Limpet Hemocyanin (KLH) protein, hepatitis B surface antigen (HBsAg), bovine serum albumin (BSA), tetanus foxoid (TT) e ⁇ the foxoid of diphtheria (DT).
• KLH Keyhole Limpet Hemocyanin
• HBsAg hepatitis B surface antigen
• BSA bovine serum albumin
• TT tetanus foxoid
• DT diphtheria
• the foxoid of diphtheria (DT) is preferably selected from the group consisting of CRM 197, CRM 76, CRM 228, CRM 45, CRM 9, CRM 102, CRM 103, e ⁇ from CRM 107.
• the carrier molecule is ⁇ CRM 197.
• the binding of the polypeptide according to the invention to a carrier molecule, in particular a carrier protein, can be carried out using a heterobifunctional coupling agent, such as the Ny-maleimidobutyryloxysuccinimide ester (GMBS) e. the sulfo-GMBS derivative, the m-maleimidobenzoyl-n-hydroxysuccinimide ester (MBS) and the sulfo-MBS derivative, succinimidyl 4- (N-maleimidome-hyl) cyclohexane-1-carboxylate (SMCC), a carbodiimide, bisdiazonium-benzidine (BDB) or glutaraldehyde
• GMBS Ny-maleimidobutyryloxysuccinimide ester
• MBS m-maleimidobenzoyl-n-hydroxysuccinimide ester
• SCC succinimidyl 4- (N-maleimidome-hyl) cyclo
• GMBS, MBS or SMCC are used, they are preferably attached to a cysteine (C), which if it is not present in a first, second, third, fourth or fifth sequence, or a sequence variant according to the invention may be added, in particular at its N-terminal or C-terminal end.
• C cysteine
• a cysteine is present in a first, a second, a third, a fourth or a fifth sequence, according to the invention at an undesired position
• BDB When BDB is used, it is preferably attached to a tyrosine (Y), which if it is not present in a first, a second, a third, a fourth or a fifth sequence, or a variant sequence according to the invention.
• Y tyrosine
• invention can be added, especially at its N-ferminal or C-terminal end.
• a tyrosine when a tyrosine is present in a first, second, third, fourth or fifth sequence according to the invention at an undesired position, it is possible to implement, instead, a variant sequence wherein the tyrosine is substituted by another amino acid, such as phenylalanine (F).
• F phenylalanine
• the binding of the polypeptide according to the invention to a carrier molecule can also be carried out using a peptide linker or peptide linker, which binds to the polypeptide according to the invention.
• a carrier molecule in particular a carrier protein
• peptide linker or peptide linker which binds to the polypeptide according to the invention.
• one side and the carrier molecule on the other side optionally via a heterobifunctional coupling agent as defined above.
• Such peptide bonds typically comprise from 1 to 10, especially from 4 to 6, amino acid residues.
• polypeptide according to the invention is attached to the CRM 197 carrier protein according to a construction represented by a formula selected from the group consisting of the following formulas:
• CRM 197 denotes the carrier protein
• GMB denotes N-g-maleimidobutyryl
• cyclo () indicates a lacfame-type cyclization between the side chains of amino acid residues at C-terminal and N-terminus
• - cycloS-S () indicates a disulfide bridge cyclization between the sulfhydryl groups of the cysteines present in C-ferminal and in N-terminal,
• brackets ([X] n) indicate that one or more polypeptides are attached to the carrier protein and the underlined part represents the polypeptide according to the invention, of which SEQ ID NO is indicated in the right column.
• polypeptide according to the invention consists of a sequence selected from the group consisting of SEQ ID NO: 22-50.
• the polypeptide according to the invention may be prepared by any method known in the state of the art and in particular by chemical synthesis. It is also possible to prepare it by expression of the nucleic acid according to the invention in eukaryotic or prokaryotic cells.
• the polypeptide according to the invention if necessary linked to a carrier molecule, is immunogenic, that is to say it can elicit or cause an immune reaction, in particular of humoral type, that is to say the production of antibodies by an individual, in particular a mammalian type, to which it is administered.
• the polypeptide according to the invention makes it possible to elicit an immune response directed against the PD-L1 protein, in particular anti-PD-L1 antibodies, preferably anfi-PD-L1 blocking or neutralizing antibodies, that is, that is to say that they prevent the PD-L1 protein from exerting part or all, especially at least 10%, 25%, 50%, 75% of its activity, for example measured in vitro.
• the activity of PD-L1 is preferably a binding to the PD-1 protein, which can be measured as shown in Example 2 which follows.
• the nucleic acid according to the invention is RNA or DNA, preferably DNA. It is preferred that the nucleic acid according to the invention is operatively linked to a prokaryotic and / or eukaryotic promoter sequence, in particular that of a mammal or a virus. Moreover, the nucleic acid according to the invention can be included in a vector, such as a plasmid or a virus. Antibodies, antibody fragments and aptamers
• the antibodies, antibody fragments, and aptamers according to the invention are said to be specifically directed against a polypeptide as defined above when they show essentially no binding to another polypeptide, which does not include the defined polypeptide. above, under conditions allowing the binding of the antibodies, antibody fragment, and aptamers according to the invention to the polypeptides against which they are specifically directed.
• the antibody according to the invention may be polyclonal or monoclonal, preferably monoclonal.
• the "antibody fragments" comprise at least one antigen-binding part of the antibody don ⁇ they are derived ⁇ , and its ⁇ in particular of Fab, Fab 'type, F (ab ') 2, disulfide stabilized Fv (dsFv), dimerized (diabody) V region, trimerized, tetramerized or pentamerized, single chain Fv (scFv), complementarity determining region (CDR).
• the antibodies can be of any species, in particular human, mouse, ra ⁇ , rabbit or camelide. Moreover, when they are not human, they can also be humanized, that is to say that the constant parts of these antibodies are replaced partially or entirely by corresponding human constant parts.
• the antibodies according to the invention can be obtained by immunization of an animal using a polypeptide according to the invention according to techniques well known to those skilled in the art.
• aptamers are nucleic acids, particularly RNAs, capable of binding specifically to a molecular target, such as a protein. Aptamers can in particular be obtained by using the SELEX technique well known to those skilled in the art, from the polypeptides according to the invention.
• the disease linked or due to the expression of the PD-L1 protein or of the PD-1 protein according to the invention is a cancer or an infectious disease.
• the disease linked to or due to PD-L1 protein or PD-1 protein is selected from the group consisting of: - unresectable metastatic melanoma, advanced non-small-cell metastatic lung carcinoma (NSCLC), advanced metastatic NSCLC progressing particularly with or after platinum-based chemotherapy, advanced renal cell carcinoma, including metastatic carcinoma, and urothelial carcinoma locally advanced or metastatic in particular not responding to chemotherapy based on platinum derivatives, prostate cancer, breast cancer, colorectal cancer, or any other cancer where the PD-1 / PD-L1 axis is involved in suppressing an anti-tumor immune response
• bacterial infections such as pneumonia, meningitis, toxic shock syndrome, food poisoning, gastritis, ulcers, gonorrhea, boils, abscesses, impetigo, ear infections, tonsillitis, infections the urinary tract and genital infections broncho-pulmonary,
• viral infections such as influenza, measles, hepatitis B, hepatitis C, infections with the human immunodeficiency virus (HIV), infections with herpes virus such as cytomegalovirus or Epsfein-Barr virus, herpes, e infections with human papillomavirus (HPV), e ⁇
• HAV human immunodeficiency virus
• herpes virus such as cytomegalovirus or Epsfein-Barr virus
• HPV human papillomavirus
• fungal infections such as blasfomycosis, coccidioiodomycosis, rhisfoplamoseja paracoccidioiodomycosis, candidiasis, cryptococcosis, aspergillosis, mucomycosis and pneumocystosis.
• the person or individuals according to the invention are animals, preferably mammals or marsupials, more preferably humans, horses, cattle, pigs, sheep, goats, camels, camels, dogs or dogs. cats, most preferably humans.
• the polypeptide according to the invention is derived from a PD-L1 protein belonging to the same species as the individual in which the polypeptide is to be used or administered.
• the polypeptide, the pharmaceutical composition, the drug or the product according to the invention is administered or in a form that can be administered orally, mucosally, in particular sublingually, parenterally, intraperitoneal, transcutaneous, intradermal, subcutaneous, intramuscular, intravenous or intra-arterial.
• the polypeptide according to the invention may be administered at doses ranging for example from 1 ng to 1 g, preferably from 1 g to 1 mg.
• a "pharmaceutically acceptable vehicle” includes all of the compounds, including excipients, that can be administered to an individual in conjunction with a pharmacological active ingredient.
• the polypeptide according to the invention may be combined or combined with an adjuvant, or the pharmaceutical composition, the drug or the product according to the invention may comprise an adjuvant.
• the adjuvant can be of any type suitable for increasing the immune response of an individual, animal or human, to the administration of a polypeptide. It can thus be complete or incomplete Freund's adjuvant, Monfanide ISA 51 VG, aluminum hydroxide, aluminum phosphate or calcium phosphate, for example; Monfanide ISA 51 VG and aluminum or aluminum phosphate hydroxides being preferred.
• the adjuvant may be combined with the polypeptide according to the invention by carrying out a 1/1 mixture by volume of an adjuvant solution and a solution comprising the polypeptide.
• other therapy designates a pharmacological therapy with at least one other compound different from the polypeptide according to the invention or a non-pharmacological therapy such as, for example, radiotherapy, in particular anti-cancer therapy.
• a non-pharmacological therapy such as, for example, radiotherapy, in particular anti-cancer therapy.
• the other compound that is useful for eliciting an immune response directed against the PD-L1 protein according to the invention can in particular be a polypeptide different from that of the invention, derived from the PD-L1 protein or a polypeptide derived from the PD-L1 protein.
• the other compound intended for the prevention or treatment of a disease linked or due to the expression of the PD-L1 protein or of the PD-1 protein, in particular during a cancer or a infectious disease can be an anticancer chemotherapy compound, an anti-cancer immunotherapy compound, for example a monoclonal antibody, an antibiotic, an antiviral, especially of the interferon type, or an antimycotic agent.
• the polypeptide according to the invention can be combined with other antigens intended to elicit an immune response. against a different target of the PD-L1 protein, for example the PD-1 protein. This type of combination is useful for the preparation of multivalent vaccines.
• the expression "in combination” or “combination product” means that the polypeptide as defined above and the other compound as defined above can be combined within one and the same pharmaceutical composition or the same drug, e ⁇ therefore be administered together, or be administered separately, that is to say, according to separate routes of administration and / or separate administration regimes, subject to when they are administered separately the periods of prophylactic or therapeutic activity of the polypeptide as defined above e ⁇ of the other compound as defined above overlap in whole or in part.
• the polypeptide as defined above when the polypeptide and the other compound are administered separately, the polypeptide as defined above will preferably be administered within 24 hours, more preferably within 2 hours, and still more preferably within 1 hour. Depending on the administration of the other compound as defined above, its administration may be continued in the following days. Conversely, the other compound as defined above will preferably be administered within 24 hours, more preferably within 2 hours, even more preferably within 1 hour, following administration of the polypeptide as defined above. , e ⁇ son administration will eventually be continued on the following days. In another preferred embodiment of the invention, when the polypeptide as defined above and the other compound as defined above are administered separately, they are administered essentially simultaneously.
• the horizontal line at 0.2 OD unit represents the significance threshold.
• Example 1 Recognition of the whole human PD-L1 protein fhPD-LI) by sera of mice immunized with peptides derived from hPD-LI.
• peptides derived from the human PD-L1 protein were synthesized chemically, cyclized by adding cysteines at their ends and then forming disulfide bridges. They were then coupled to a carrier protein, CRM 197 (C-Reactive Material 197), using the GMBS coupling agent.
• CRM 197 C-Reactive Material 197
• amino acid residues are annotated from the sequence of the PD-protein
• Example 2 Neutralization of the biological activity of human PD-L1 with antibodies purified from the serum of rabbits immunized with the peptides derived from
• Neutralizing capacity of purified IgG from rabbit serum (n 4 / group) respectively immunized with peptides PPV-09-01, PPV-09-02, PPV-09-03, PPV-09-04, PPV- 09-05 or PPV-09-06, was evaluated in a PD-1 / PD-L1 interaction neutralizing cell (Promega, J 1250). This is based on the interaction between 2 cell lines:
• CHO-K1 cell line expressing the human PD-L1 gene and a surface protein making it possible to activate the TCRs in an antigen-dependent manner.
• Plating J l: Inoculation of a 96-well flat-bottomed plate treated for cell culture with CHO-K1 cells. Incubate the plate for 20h at 37 ° C.

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