Anti-NGF vaccine composition

Abstract

The present invention relates to a polypeptide comprising, or consisting of, - a first sequence consisting of at least eight consecutive amino acids selected from the sequence of amino acids from position 122 to 136 of the β-subunit of an NGF protein and up to 30 consecutive amino acids selected from the complete sequence of the β-subunit of an NGF protein; and / or - a second sequence consisting of at least eight consecutive amino acids selected from the sequence of amino acids from position 148 to 158 of the β-subunit of an NGF protein and up to 30 consecutive amino acids selected from the complete sequence of the β-subunit of an NGF protein.

Description

Technical Field

[0001] The present invention relates to peptides or polypeptides useful for inducing an immune response directed against nerve growth factor (NGF).

Background Art

[0002] Chronic pain (CRNCP) is a major unmet medical need and there is a pressing need for safer and more effective drugs. Among the most common forms or causes of chronic pain are i) neuropathic pain (NP) associated with nerve damage in the peripheral or central nervous system; and ii) osteoarthritis (OA), a progressive degeneration of articular cartilage characterized by inflammation and pain.

[0003] There is no completely effective treatment for the widespread chronic pain associated with these disorders, and current therapies (NSAIDs and opioids) cause serious unwanted side effects.

[0004] In recent years, the NGF ligand-receptor system has emerged as a novel target with significant therapeutic potential for NP and OA. Indeed, NGF and its receptors (TrkA and p75 NTR ) are thought to be major regulators of both the neuropathic and inflammatory elements of pain. Excessive levels of NGF caused by tissue damage or other factors result in rapid sensitization of nociceptors, decreasing the pain threshold and increasing the intensity of pain experienced (e.g., allodynia and hyperalgesia). Genetic studies in humans have implicated mutations in the genes of the TrkA receptor and NGF itself in causing congenital forms of pain insensitivity (hereditary sensory and autonomic neuropathy type IV, HSAN IV and HSAN V, respectively), validating NGF as a target. Thus, blocking the NGF signaling system is a rational and well-validated approach to pain management.

[0005] In fact, the potent analgesic efficacy of certain anti-NGF monoclonal antibodies (mAbs), such as tanezumab (Eli Lilly and Company) and facinumab (Mirati Therapeutics), has been demonstrated in various preclinical models of CRNCP, from NP models to inflammatory models (cancer pain and osteoarthritis, and interstitial cystitis, respectively), and has also been demonstrated in clinical trials (Gondal et al. (2022) Drugs Today (Barc.) 58:187-200).

[0006] Other pathological conditions are also linked to NGF. In fact, NGF is widely expressed in lung tissue. In various microenvironments, NGF contributes to the onset and development of lung diseases by altering protein expression levels and acting on cellular function. Therefore, NGF may be involved in certain underlying mechanisms in pulmonary fibrosis (PF), coronavirus 2019 disease (COVID-19), pulmonary hypertension (PH), asthma, chronic obstructive pulmonary disease (COPD), and lung cancer. In particular, NGF is an important mediator in the neuroimmunological mechanisms of asthma, exacerbating inflammation and airway remodeling (Li et al. (2009) Respirology 14:60~68). Serum NGF levels are also extremely high in COPD subjects (Stabile et al. (2016) J. Biol. Chem. 397:157~163).

[0007] In tumor tissue, NGF stimulates angiogenesis (including in non-small cell lung cancer (NSCLC)) (Lu et al. (2014) J. Huazhong Univ. Sci. Technol. Med. Sci. 34:359~362). NGF is also present in breast cancer and is produced in tumor cells but not in normal epithelial cells, so it can be used not only for diagnosis but also as a treatment target. Therefore, the use of anti-NGF therapy may form the basis of future treatment strategies for the aforementioned lung diseases.

[0008] However, monoclonal antibodies have several well-known limitations: High cost, the need for frequent re-administration, and the occurrence of an immune response directed towards the administered monoclonal antibody. It has.

[0009] Therefore, the objective of the present invention is to overcome these drawbacks. [Overview of the Initiative]

[0010] This invention arose from the inventors' unexpected discovery that polypeptides derived from the sequences of amino acids 122 to 136 and 148 to 158 of the β-subunit of human NGF protein (hNGF) can produce antibodies directed against hNGF protein, particularly neutralizing antibodies.

[0011] Therefore, the present invention is - In particular, a first sequence consisting of at least eight consecutive amino acids selected from the sequence of amino acids 122 to 136 of the β-subunit of the NGF protein and up to 30 consecutive amino acids selected from the complete sequence of the β-subunit of the NGF protein, or a variant sequence showing at least 75% identity with the first sequence; and / or - A second sequence consisting of at least eight consecutive amino acids selected from the sequence of amino acids 148 to 158 of the β-subunit of the NGF protein and up to 30 consecutive amino acids selected from the complete sequence of the β-subunit of the NGF protein, or a variant sequence having at least 75% identity with the second sequence. A polypeptide comprising or consisting thereof, However, polypeptides are different from the β-subunit of NGF protein, and do not consist of a continuous portion of more than 30 amino acids from the β-subunit of NGF protein, and However, this relates to polypeptides in which the polypeptides, consisting of variant sequences of the first and second sequences, can induce an immune response directed towards NGF proteins.

[0012] In preferred embodiments, the present invention is more particularly, - Sequence IDs 1, 22, 24, 26, 28, or 29, in particular a first sequence consisting of sequence ID 1 or 28, or a variant sequence having at least 75% identity with the first sequence; and / or - Sequence IDs 2, 23, 25, or 27, in particular a second sequence consisting of sequence ID 2 or 27, or a variant sequence having at least 75% identity with the second sequence. This relates to polypeptides containing or consisting of the following.

[0013] The present invention also relates to nucleic acids or their complements that encode the polypeptides defined above.

[0014] The present invention also relates, in particular, to pharmaceuticals for human or veterinary use, especially for use as a vaccine: - At least one polypeptide as defined above, or - Also relating to at least one nucleic acid as defined above. In certain embodiments of the present invention, the pharmaceuticals, in particular vaccines, as defined above, also include at least one other compound intended to prevent or treat diseases linked to or caused by NGF proteins.

[0015] The present invention also includes, optionally, at least one pharmaceutically acceptable vehicle, as an active substance: - At least one polypeptide as defined above, or - The invention also relates to a pharmaceutical composition, in particular a vaccine, comprising at least one nucleic acid as defined above.

[0016] In certain embodiments of the present invention, the pharmaceutical composition defined above, in particular the vaccine composition, also includes at least one other compound intended to prevent or treat diseases linked to or caused by NGF proteins.

[0017] The present invention also relates to the use of polypeptides as defined above for the preparation of antibodies, antibody fragments, or aptamers.

[0018] The present invention also relates to a method for preparing an antibody, antibody fragment, or aptamer, comprising the steps of administering the polypeptide defined above to an antibody-producing organism, or performing affinity selection of an antibody, antibody fragment, or aptamer that binds to the polypeptide defined above.

[0019] The present invention also relates to an anti-NGF antibody, antibody fragment, or aptamer specifically directed to the polypeptide defined above, wherein the polypeptide comprises two or fewer amino acid residues in addition to the first sequence, the second sequence, or each variant sequence thereof.

[0020] The present invention also relates to antibodies, antibody fragments, or aptamers as defined above for use as pharmaceuticals. In certain embodiments of the present invention, the pharmaceuticals as defined above also include at least one other compound for the prevention or treatment of diseases relating to or caused by NGF proteins.

[0021] The present invention also relates to pharmaceutical compositions comprising, optionally, a pharmaceutically acceptable vehicle, the antibody, antibody fragment, or aptamer defined above as an active ingredient. In certain embodiments of the present invention, the pharmaceutical compositions defined above also include at least one other compound intended to prevent or treat diseases linked to or caused by NGF proteins.

[0022] The present invention also relates to the use of the polypeptide defined above, the nucleic acid (RNA or DNA) defined above, or the pharmaceutical composition defined above for use in a method for inducing an immune response directed against NGF protein in an individual. In certain embodiments of the present invention, the polypeptide, nucleic acid or pharmaceutical composition is used in combination with at least one other compound useful for inducing an immune response directed against NGF protein.

[0023] The present invention also relates to a method for inducing an immune response directed against NGF protein in an individual, the method comprising administering to the individual an effective amount of the polypeptide defined above, the nucleic acid defined above, or the pharmaceutical composition defined above. In certain embodiments of the present invention, the polypeptide, nucleic acid or pharmaceutical composition is administered in combination with at least one other compound useful for inducing an immune response directed against NGF protein.

[0024] The present invention also relates to the use of the polypeptide defined above or the nucleic acid defined above for the preparation of a medicament for inducing an immune response directed against NGF protein in an individual. In certain embodiments of the present invention, the medicament also comprises at least one other compound useful for inducing an immune response directed against NGF protein.

[0025] The present invention also relates to the use of the polypeptide defined above, the nucleic acid defined above, the pharmaceutical composition defined above, or the antibody, antibody fragment or aptamer defined above for use in a method for preventing or treating a disease related to or caused by NGF protein in an individual. In certain embodiments of the present invention, the polypeptide, nucleic acid, pharmaceutical composition, or antibody, antibody fragment or aptamer is used in combination with at least one other therapy for preventing or treating a disease related to NGF protein.

[0026] The present invention also relates to a method for preventing or treating a disease relating to or caused by NGF proteins in an individual, comprising administering to the individual an effective amount of the polypeptide, nucleic acid, pharmaceutical composition, or antibody, antibody fragment, or aptamer defined above. In certain embodiments of the present invention, the method comprises at least one further therapeutic method for preventing or treating a disease relating to NGF proteins.

[0027] The present invention also relates to the use of the polypeptides, nucleic acids, or antibodies, antibody fragments, or aptamers defined above for the preparation of pharmaceuticals for preventing or treating diseases related to or caused by NGF proteins in an individual. In certain embodiments of the present invention, the pharmaceutical comprises at least one other compound intended to prevent diseases linked to or caused by NGF proteins.

[0028] The present invention also provides combination products for simultaneous, separate, or sequential use in the prevention or treatment of diseases linked to or caused by NGF proteins, as follows: - The polypeptide or nucleic acid as defined above, and - At least one other compound intended to prevent or treat diseases linked to or caused by NGF proteins This also applies to products that contain [the specified substance].

[0029] Description of the Invention As a preliminary point, the term "comprising" means "including," "containing," or "encompassing," meaning that when an object "comprises" one or more elements, elements other than those stated may also be included in the object. On the other hand, the expression "consisting of" means "constituted by," meaning that when an object "consists of" one or more elements, the object must not contain elements other than those stated.

[0030] Polypeptide Definition of NGF protein NGF protein refers to nerve growth factor. It is also known as NGF, β-HSAN5, or NGFB, and is well known to those skilled in the art. NGF is a multimeric protein containing α, β, and γ subunits, and generally has the stoichiometry α2β2γ2. The β subunit of NGF, or β-NGF, is responsible for catalytic activity. NGF is often used synonymously with β-NGF.

[0031] Preferred species and sequence of the β-subunit of NGF protein Preferably, the β-subunit of the NGF protein according to the present invention is selected from the group consisting of the β-subunit of human NGF protein, the β-subunit of mouse NGF protein, the β-subunit of rat NGF protein, the β-subunit of rabbit NGF protein, the β-subunit of ape NGF protein, the β-subunit of dog NGF protein, the β-subunit of cat NGF protein, the β-subunit of horse NGF protein, the β-subunit of pig NGF protein, the β-subunit of sheep NGF protein, and the β-subunit of dromedary camel NGF protein. Particularly preferably, the NGF protein is the β-subunit of human NGF protein (hNGF) or dog NGF protein.

[0032] Preferably: - The β-subunit of human NGF protein (hNGF) is as described in the UniProt / Swissprot database under reference number P01138, and consists of sequence number 3. - The β-subunit of the mouse NGF protein (mNGF) is as described in the UniProt / Swissprot database under reference number P01139, and consists of sequence number 4. - The β-subunit of the rat NGF protein is as described in the NCBI Genbank database under reference number NP_001263984.1, or as described in the Uniprot / SwissProt database under reference number P25427, and consists of sequence number 5 or 18. - The β-subunit of the rabbit NGF protein is as described in the NCBI Genbank database under reference number XP_051712092.1, and consists of sequence number 6. - The β-subunit of the ape NGF protein is as described in the Genbank database under reference number NP_001012439.1, or as described in the Uniprot / SwissProt database under reference number Q9N2F1, and consists of sequence number 7 or 19. - The β-subunit of the canine NGF protein is as described in the Uniprot / SwissProt database under reference number A0A8I3PYI3, and consists of sequence number 8. - The β-subunit of the feline NGF protein is as described in the Genbank database under reference number XP_004001166.2, and consists of sequence number 9. - The β-subunit of the equine NGF protein is as described in the Genbank database under reference number XP_001496237.2, and consists of sequence number 10. - The β-subunit of the porcine NGF protein is as described in the Genbank database under reference number XP_020945655.1, and consists of sequence number 11. - The β-subunit of the sheep NGF protein is as described in the UniProt / Swissprot database under reference number A0A481MW93, or as described in the Genbank database under reference number XP_060257455.1, and consists of sequence number 12 or 20. - The β-subunit of the dromedary camel NGF protein is as described in the UniProt / Swissprot database under reference number A0A5N4DT36, and consists of sequence number 13.

[0033] Numbering of amino acid residues As understood herein, the amino acid residue numbering of the β-subunit of an NGF protein begins with the first amino acid residue forming the N-terminus of the β-subunit of a full-length NGF protein encoded by the open reading frame of the NGF β-subunit (β-NGF) gene, usually methionine (M), i.e., including its signal peptide and constitutive pro-NGF propeptide. Furthermore, the amino acid residue numbering of the β-subunit of an NGF protein as used herein is defined by referring to the β-subunit of a human NGF protein. Therefore, determining the amino acid residues of the β-subunit of the NGF protein corresponding to the position number referenced according to the present invention is easy for those skilled in the art: this only requires aligning the sequence of the β-subunit of the NGF protein for which the amino acid residues corresponding to the position number are to be determined with the sequence of the β-subunit of the human NGF protein, in particular sequence number 3, thereby optimizing the identity percentage between the two aligned sequences, and then identifying the amino acid residues corresponding to the position number sought to align with the amino acid residues of the sequence of the β-subunit of the human NGF protein having this position number.

[0034] First and second sequences Preferably, the first sequence and the second sequence each consist of at least 9, 10, 11, and 12 consecutive amino acids selected from one of the sequences extending from amino acid positions 122 to 136 or from amino acid positions 148 to 158 of the β-subunit of the NGF protein, or each consists of at least the sequence extending from amino acid positions 122 to 136 or from amino acid positions 148 to 158 of the β-subunit of the NGF protein.

[0035] Preferably, the first and second sequences according to the present invention consist of up to 29, 28, 27, 26, 25, 24, 23, 22, 21, and 20 consecutive amino acids selected from the complete sequence of the β-subunit of the NGF protein, or they consist of up to the sequences from positions 122 to 136 and 148 to 158 of the β-subunit of the NGF protein, respectively.

[0036] Preferably, the first and second sequences according to the present invention consist of (i) Sequence IDs 1, 22, 24, 26, 28, or 29, in particular Sequence ID 1 or 28, and (ii) Sequence IDs 2, 23, 25, or 27, in particular Sequence ID 2 or 27.

[0037] Sequence IDs 1 and 2 represent, in particular, the 122-136 and 148-158 positions of the β-subunit of the human NGF protein represented by Sequence ID 3, respectively.

[0038] [Table 1]

[0039] Sequence ID 22 specifically represents the portion of the β-subunit of the mouse or rat NGF protein represented by Sequence IDs 4, 5, or 18, from position 122 to 136.

[0040] Sequence ID 23 specifically represents the portion of the β-subunit of the mouse or rat NGF protein represented by Sequence IDs 4, 5, or 18, from position 148 to 158.

[0041] Sequence IDs 24 and 25 represent, in particular, the 122-136 and 148-158 portions of the β-subunit of the rabbit NGF protein represented by Sequence ID 6, respectively.

[0042] Sequence ID 26 specifically represents the portion of the β-subunit of the ape (chimpanzee) NGF protein represented by Sequence ID 7 or 19, specifically from position 122 to 136.

[0043] Sequence ID 27 specifically represents the portion of the β-subunit of the ape (chimpanzee), dog, cat, horse, pig, or sheep NGF protein represented by Sequence ID 7 or 19, 8, 9, 10, 11, 12, or 20, from position 148 to 158, or specifically the portion of the β-subunit of the dromedary camel NGF protein represented by Sequence ID 13.

[0044] Sequence ID 28 specifically represents the portion of the β-subunit of the canine, cat, horse, pig, or sheep NGF protein represented by Sequence IDs 8, 9, 10, 11, 12, or 20, from position 122 to 136.

[0045] Sequence ID 29 specifically represents the β-subunit of the dromedary camel NGF protein represented by Sequence ID 13, specifically the portion from positions 122 to 137.

[0046] Variant array The variant sequence according to the present invention has at least 75% identity with the first or second sequence shown above, and preferably at least 80%, 85%, 90%, 95%, or 98% identity with the first or second sequence.

[0047] As understood herein, the percentage of identity between two peptide sequences can be determined by performing optimal alignment over the entire length of the sequences, finding the number of alignment positions where both sequences have identical amino acids, and dividing this number by the total number of amino acids in the longer of the two sequences. Optimal alignment is the one that maximizes the percentage of identity between the two sequences.

[0048] Preferably, the variant sequence according to the present invention has at least 75%, 80%, 85%, 90%, 95%, or 98% identity with sequence numbers 1, 22, 24, 26, 28, or 29 or sequence numbers 2, 23, 25, or 27.

[0049] The variant sequence according to the present invention is such that the polypeptide comprising the variant sequence must be able to induce an immune response directed against the NGF protein; that is, when a peptide (optionally linked to a carrier molecule, in particular a carrier protein, e.g., KLH (keyhole limpet hemocyanin)) is administered to an animal such as a mouse, rat, or rabbit, it may be cyclized by the formation of a disulfide crosslink between at least one cysteine, optionally within the peptide and / or after the addition of one or two cysteines at its N-terminus and / or C-terminus, it induces the production of antibodies directed against NGF, in particular against NGF of the same species to which the variant sequence having the highest percentage of identity belongs. Those skilled in the art are well aware of methods for determining whether an antibody is directed against NGF, particularly by performing an ELISA test. Preferably, the antibody induced by the administration of the conjugate peptide is a blocking or neutralizing antibody, that is, it prevents the NGF protein from exhibiting all or part of its activity, particularly at least 10%, 25%, 50%, or 75%, as measured in vitro.

[0050] Polypeptide length The polypeptides according to the present invention preferably contain up to 200, 150, 100, 90, 80, 70, 60, 50, 40, or 30 amino acid residues. The polypeptides according to the present invention are distinct from the β-subunit of the NGF protein and do not consist of a portion of the β-subunit of the NGF protein with more than 30 consecutive amino acid residues. As will be understood by those skilled in the art, this does not preclude the fact that these portions consist of two or more portions of the β-subunit of the NGF protein with up to 30 consecutive amino acid residues, unless they are arranged to reconstitute a portion of the β-subunit of the NGF protein with more than 30 consecutive amino acids.

[0051] As will be apparent to those skilled in the art, the polypeptide according to the present invention may include several repeat sequences, for example, 2, 3, 4, 5, 10, or 20 repeat sequences, in each of the first sequence, the second sequence, and the variant sequence according to the present invention.

[0052] The first array, the second array, and the array to be added to the variant array. Furthermore, the polypeptide according to the present invention may also include sequences that do not originate from the β-subunit of the NGF protein, in particular sequences that include epitopes belonging to other proteins, thereby making it possible to induce or produce an immune response directed towards these other proteins.

[0053] Furthermore, the immunogenicity of the polypeptide according to the present invention can be enhanced by including an exogenous, preferably universal, T-epitope (one or more) sequence.

[0054] The polypeptide according to the present invention may also include at least one sequence of a carrier protein, for example, a virus-like particle (VLP) described in detail in international application WO05 / 117983 relating to TNF.

[0055] Polypeptide cyclization The polypeptide according to the present invention may be cyclized or not. Preferably, the polypeptide according to the present invention is cyclized. This cyclization may be of any type known to those skilled in the art. The selection of the cyclization strategy according to the present invention may take into account, in particular, the optimal antigen presentation of the epitope contained in the polypeptide according to the present invention.

[0056] Depending on the functional groups present, this cyclization of a polypeptide can occur in several different ways, for example: from its C-terminus to its N-terminus, from its N-terminus to a side chain, from a side chain to its C-terminus, or between two side chains. Among the various ways in which a polypeptide can be cyclized are lactamization, lactonization, and disulfide crosslinking. In particular, when disulfide crosslinking is formed between cysteines, i.e., between the -SH radicals of two cysteines, the cysteine ​​may already be present in the variant sequence according to the present invention or the first and second sequences according to the present invention, or may be added within these sequences as well as at the N-terminus and / or C-terminus.

[0057] Post-translational modification, amino acid analog Furthermore, the polypeptide according to the present invention may include post-translational modifications, such as glycosylation, methylation, particularly acylation with fatty acids, or phosphorylation. In particular, the N-terminus of the polypeptide according to the present invention may be acetylated, and the C-terminus may be modified by amidation.

[0058] The polypeptide according to the present invention may also include one or more analogs or derivatives of amino acids, including non-natural or non-standard amino acids.

[0059] Carrier molecule Preferably, the polypeptide according to the present invention is further attached to or linked to a carrier molecule, particularly a carrier protein, by covalent bonds.

[0060] In particular, the carrier molecules may be keyhole limpet hemocyanin (KLH) protein, hepatitis B surface antigen (HBsAg), bovine serum albumin (BSA), tetanus toxoid (TT), and diphtheria toxoid (DT).

[0061] The diphtheria toxoid (DT) according to the present invention is preferably selected from the group consisting of CRM 197, CRM 176, CRM 228, CRM 45, CRM 9, CRM 102, CRM 103, and CRM 107.

[0062] A particularly preferred carrier molecule is CRM 197.

[0063] The linkage of polypeptides according to the present invention to carrier molecules, particularly carrier proteins, can be achieved using the following: heterobifunctional coupling agents, such as N-γ-maleimidobutyryl-oxysuccinimide esters (GMBS) and sulfo-GMBS derivatives, m-maleimidobenzoyl-n-hydroxysuccinimide esters (MBS) and sulfo-MBS derivatives, succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), carbodiimide, bisdiazonium-benzidine (BDB), or glutaraldehyde.

[0064] When glutaraldehyde is used, it is preferably added to lysine (K), and if it is not present in the first or second sequence or variant sequence according to the present invention, it can be added particularly to the N-terminus or C-terminus. On the other hand, if lysine is present in an undesirable position in the portion according to the present invention, it is preferable to use a different conjugation method.

[0065] When used, BDB is preferably attached to tyrosine (Y), and can be added to the N-terminus or C-terminus, in particular, if it is not present in the first or second sequence or variant sequence according to the present invention. Furthermore, if tyrosine is present in an undesirable position in the portion according to the present invention, a variant sequence in which tyrosine is substituted with another amino acid such as phenylalanine (F) can be used instead.

[0066] GMBS, MBS, or SMCC, when used, are preferably added to cysteine ​​(C), and can be added to the N-terminus or C-terminus of the sequence, particularly when they are not present in the first or second sequence or variant sequence according to the present invention. Furthermore, if cysteine ​​is present in an undesirable position in the first or second sequence according to the present invention, a variant sequence in which cysteine ​​is substituted with another amino acid such as serine can be used instead.

[0067] Particularly preferred is a polypeptide according to the present invention, selected from the group consisting of the following formulas: [Table 2] (In the formula: - CRM197 refers to the carrier protein, - Acetyl+ indicates that the N-terminus is acetylated. - CycloSS() represents disulfide bridging cyclization between the sulfhydryl groups of the C-terminus and N-terminus cysteine. - Ahx represents an amino-hexanoic acid type unnatural amino acid, - Square brackets ([X] n ) indicates that one or more polypeptides are attached to the carrier protein. (The underlined bolded portion represents the first or second arrangement according to the present invention.) It is attached to the CRM197 carrier protein according to the configuration represented by [the symbol].

[0068] Polypeptide preparation The polypeptides according to the present invention can be prepared by any method known in the current state of the art, in particular by chemical synthesis. The polypeptides according to the present invention can also be prepared by the expression of nucleic acids according to the present invention in eukaryotic or prokaryotic cells.

[0069] Polypeptide activity The polypeptides according to the present invention, when linked to a carrier molecule as necessary, are immunogenic, that is, they can induce or trigger humoral immune responses, in other words, the production of antibodies of the type of mammal being administered to an individual. In particular, the polypeptides according to the present invention can induce an immune response directed against NGF proteins, in particular anti-NGF antibodies, preferably blocking or neutralizing anti-NGF antibodies, that is, these antibodies prevent NGF proteins from exhibiting all or part of their activity, for example, measured in vitro, particularly at least 5%, 10%, 25%, 50%, or 75%.

[0070] nucleic acid The nucleic acid according to the present invention is RNA or DNA, preferably DNA. Preferably, the nucleic acid according to the present invention is ligated to act on a promoter sequence of a prokaryote and / or eukaryote, in particular a mammal or a virus. Alternatively, the nucleic acid according to the present invention may be contained in a vector, such as a plasmid or a virus.

[0071] Antibodies, antibody fragments, and aptamers The antibodies, antibody fragments, and aptamers according to the present invention are said to be specifically directed to the polypeptides defined above if, under conditions in which the antibodies, antibody fragments, and aptamers according to the present invention can bind to the polypeptides to which they are specifically directed, they do not substantially bind to other polypeptides that do not contain the polypeptides defined above.

[0072] The antibody according to the present invention may be a polyclonal antibody or a monoclonal antibody, preferably a monoclonal antibody. Furthermore, as understood herein, “antibody fragment” comprises at least one antigen-binding region of the derived antibody, in particular of the types Fab, Fab', F(ab')2, disulfide-stabilized Fv(dsFv), dimerized (diabody), trimerized, tetramerized or pentamerized V region, single-chain Fv(scFv), and complementarity-determining region (CDR).

[0073] Antibodies can be from any species, including humans, mice, rats, rabbits, or camelids. Furthermore, if they are not human, they can be humanized, meaning that the constant portion of these antibodies is partially or completely replaced by the corresponding human constant portion.

[0074] The antibodies according to the present invention can be obtained by immunizing animals with the polypeptide according to the present invention using methods well known to those skilled in the art.

[0075] As understood herein, an aptamer is a nucleic acid, particularly RNA, that can specifically bind to a molecular target, such as a protein. In particular, aptamers can be obtained from polypeptides according to the present invention using the SELEX method, which is well known to those skilled in the art.

[0076] therapeutic use disease Various diseases, both human and veterinary, particularly chronic pain, can be treated by the present invention. Other examples of conditions or disorders include asthma, psoriasis, osteoarthritis, and arthritis.

[0077] The present invention is often useful in treating or preventing chronic pain or recurrent generalized or localized pain related to NGF activity.

[0078] The present invention can be used for the prevention, symptomatic treatment, or treatment of the disease itself. The present invention can be used for the treatment of asthma and other disorders associated with airway hyperresponsiveness, which are typically characterized by episodes of cough and / or respiratory problems.

[0079] The present invention is also useful in the management of other inflammatory conditions, such as chronic inflammatory bowel diseases including multiple sclerosis, colitis, Crohn's disease, or ulcerative colitis, cystitis, eczema, contact dermatitis, arthritis including chronic arthritis and rheumatoid arthritis, and psoriasis.

[0080] The present invention is also useful for treating diseases that may be associated with increased NGF levels, including, for example, osteoarthritis and lupus, as well as chronic obstructive pulmonary disease (COPD), lung cancer, hyperalgesia, chronic pain, and widespread or localized recurrent pain.

[0081] individual The one or more individuals according to the present invention are animals, preferably mammals or marsupials, more preferably humans, horses, cattle, pigs, sheep, goats, dromedary camels, dogs, or cats, most preferably humans or dogs.

[0082] Administration Preferably, the polypeptide, pharmaceutical composition, pharmaceutical, or product according to the present invention is administered in a form that can be administered orally, mucously, particularly sublingually, parenterally, intraperitoneally, percutaneously, intradermally, subcutaneously, intramuscularly, intravenously, or intraarterially.

[0083] dose In the context of the present invention, the polypeptide according to the present invention can be administered in doses ranging from 1 ng to 1 g, preferably from 1 μg to 1 mg.

[0084] Pharmacologically acceptable vehicles As used herein, “pharmaceutically acceptable vehicle” includes all compounds that contain excipients that can be administered to an individual together with the pharmacologically active ingredient.

[0085] Adjuvant Furthermore, particularly when used in vaccines or prophylaxis, the polypeptides according to the present invention can be associated with or combined with adjuvants, or the pharmaceutical compositions, pharmaceuticals, or products according to the present invention may contain adjuvants. The adjuvant can be any type suitable for increasing the immune response of an individual, animal, or human in relation to the administration of the polypeptide. Examples include complete or incomplete Freund's adjuvant, Montanide ISA51 VG, alum, or calcium phosphate, with Montanide ISA51 VG and alum being preferred. The adjuvant can be combined with the polypeptide according to the present invention by preparing a mixture of the adjuvant solution and the polypeptide-containing solution in a 1:1 volume ratio.

[0086] Other treatments As used herein, the expression “other treatments” refers to pharmacological treatments using at least one other compound different from the polypeptide according to the present invention, or non-pharmacological treatments such as the use of anti-IgGE antibodies in the treatment of asthma.

[0087] Other compounds Another compound useful for inducing an immune response directed against the NGF protein according to the present invention may be a polypeptide different from the polypeptide of the present invention, which is also derived from the NGF protein.

[0088] Furthermore, particularly when used in vaccine formulations or included in vaccines or vaccine compositions, the polypeptides according to the present invention can be combined with other antigens intended to induce an immune response to targets different from NGF proteins. This type of combination is useful for preparing polyvalent vaccines.

[0089] As used herein, the expressions “in combination” or “combination product” mean that the polypeptides defined above and the other compounds defined above may be administered together in combination in the same pharmaceutical composition or medicine, or that they may be administered separately, i.e., by separate routes of administration and / or separate administration regimens, provided that when administered separately, the durations of the prophylactic or therapeutic activity of the polypeptides defined above and the other compounds defined above overlap in whole or in part.

[0090] Therefore, the polypeptide defined above, when administered separately, is preferably administered within 24 hours, more preferably within 2 hours, and even more preferably within 1 hour following the administration of the other compound defined above, and the administration may be continued on the following day or later. Conversely, the other compound defined above is preferably administered within 24 hours, more preferably within 2 hours, and even more preferably within 1 hour following the administration of the polypeptide defined above, and the administration may be continued on the following day or later. In another preferred embodiment of the present invention, when the polypeptide defined above and the other compound defined above are administered separately, they are administered substantially simultaneously.

[0091] [Table 3] TIFF2026518416000004.tif248169 TIFF2026518416000005.tif242169

[0092] The present invention is further illustrated by the following drawings and non-limiting embodiments. [Brief explanation of the drawing]

[0093] [Figure 1]This figure shows the anti-hNGF antibody titer of 50 (y axis) measured from the serum of 6 SWISS mice immunized with the conjugate peptide PPV-11-13 (derived from Sequence ID No. 1 of Example 1). The x axis corresponds to the time (unit: days) when serum was collected before and after the first immunization (day 0 (D0)). [Figure 2] This figure shows the anti-hNGF antibody titer of 50 (y axis) measured in serum from six SWISS mice immunized with the conjugate PPV-11-18 peptide (derived from Sequence ID No. 2 in Example 2). The x axis corresponds to the time (in days) when serum was collected before and after the first immunization (Day 0 (D0)). [Figure 3] This figure shows the anti-hNGF antibody titer of 50 (y axis) measured from the serum of five rabbits immunized with the conjugate peptide derived from Sequence ID No. 1 of Example 3, as a function of the time of serum collection (unit: days) (x axis). The red arrows indicate the immunization days (day 0 (D0), day 28 (D28), and day 119 (D119)). [Figure 4] This figure shows the inhibition percentage of 8 ng / mL hNGF against its receptor hTrkA by purified antibodies (1 / 2 dilution) from five rabbits immunized with the conjugate peptide derived from Sequence ID No. 1 of Example 3. These antibodies were measured at 45 days (D45), 80 days (D80), 110 days (D110), 136 days (D136), 180 days (D180), and 209 days (D209). [Figure 5] This figure shows the anti-hNGF antibody titer of 50 (y axis) measured from the serum of five rabbits immunized with the conjugate peptide derived from Sequence ID No. 2 from Example 4, as a function of serum collection time (unit: days) (x axis). The red arrows indicate the immunization days (day 0 (D0), day 28 (D28), and day 119 (D119)). [Figure 6]This figure shows the inhibition percentage of 8 ng / mL hNGF against its receptor hTrkA by purified antibodies (1 / 2 dilution) from five rabbits immunized with the conjugate peptide derived from Sequence ID No. 2 of Example 4. These antibodies were present at day 45 (D45), day 80 (D80), day 110 (D110), day 136 (D136), day 180 (D180), and day 209 (D209). [Figure 7] This figure shows the evaluation of the analgesic effect of β-hNGF-derived peptides in a rat model of monosodium iodoacetate-induced osteoarthritis. Figure 7 shows the weight balance between the ipsilateral leg (MIA) and the contralateral leg as a percentage (y-axis), calculated as [(weight loaded on the ipsilateral leg / total weight loaded on the ipsilateral and contralateral legs) × 100], measured in rats at different time points after intra-articular injection of MIA (1 mg / rat) (unit: days) (x-axis). G1: Vehicle group, rats (n=12, 6 males + 6 females) subcutaneously immunized with 0.1 mL of PBS emulsified with Montanide adjuvant® ISA 51 VG. G2: Test group, rats (n=12, 6 males + 6 females) subcutaneously immunized with 0.1 mL of the equivalent of 25 μg of peptide (SEQ ID NO: 22 from Example 5) emulsified with Montanide adjuvant® ISA 51 VG. G3: Test group, rats (n=12, 6 males + 6 females) subcutaneously immunized with 0.1 mL of a 25 μg equivalent of the peptide emulsified in Montanide adjuvant (trademark) ISA 51 VG (SEQ ID NO: 23 from Example 5). *p<0.05, **p<0.01, ***p<0.001, compared to the control group (PBS emulsion only, G1). Single-measure ANOVA followed by Tukey's post-hoc test. [Examples]

[0094] The following examples demonstrate how all of the peptides of the present invention, derived from NGF sequences of several different animals, induce the production of antibodies capable of neutralizing the activity of homogeneous or heterogeneous β-NGF proteins:

[0095] (Example 1) Recognition of the β-subunit (β-hNGF) of human NGF protein by serum from mice immunized with peptides derived from β-hNGF. A peptide containing positions 122-136 of the β-subunit of the human NGF protein (PPV-11-13, see Table 2) was synthesized by solid-phase chemical synthesis using standard methods well known to those skilled in the art, and then cyclized by forming a disulfide crosslink between two cysteine ​​units of the peptide. The peptide was then coupled to the carrier protein KLH (keyhole limpet hemocyanin) using glutaraldehyde as a coupling agent.

[0096] Swiss mice free of specific pathogens (Janvier Labs, Le Genest-Saint-Isle, France) were subcutaneously immunized with 25 μg equivalent of PPV-11-13 peptide emulsified in Montanide ISA 51 VG adjuvant (n=6). Mice received three subcutaneous injections (day 0 (D0), day 28 (D28), and day 84 (D84)).

[0097] [Table 4]

[0098] The relative amount of anti-β-hNGF antibody is evaluated in mouse serum by ELISA on days -4 (D-4 (pre-immunization)), 56 (D56), 98 (D98), and 126 (D126).

[0099] We observed that all immunized mice produced antibodies that recognized the β-hNGF protein (Figure 1). The PPV-11-13 peptide conjugate induces an immune response to the β-subunit of the human NGF protein in mice.

[0100] (Example 2) Recognition of the β-subunit (β-hNGF) of human NGF protein by serum from mice immunized with peptides derived from β-hNGF. A peptide containing positions 148-158 of the β-subunit of the human NGF protein (PPV-11-18, see Table 3) was synthesized by solid-phase chemical synthesis using standard methods well known to those skilled in the art, and then cyclized by forming a disulfide bridge between two cysteine ​​units of the peptide. The peptide was then coupled to the carrier protein CRM 197 using a maleimide coupling agent.

[0101] Specific pathogen-free SWISS mice (Janvier Labs, Le Genest-Saint-Isle, France) were subcutaneously immunized with 25 μg equivalent of PPV-11-18 peptide emulsified in Montanide ISA 51 VG adjuvant (n=6). Mice received three subcutaneous injections (day 0 (D0), day 28 (D28), and day 84 (D84)).

[0102] [Table 5]

[0103] The relative amount of anti-β-hNGF antibody is evaluated in mouse serum by ELISA on days -4 (D-4 (pre-immunization)), 56 (D56), 98 (D98), and 126 (D126).

[0104] We observed that all immunized mice produced antibodies that recognized the β-hNGF protein (Figure 2). The PPV-11-18 peptide conjugate induces an immune response to the β-subunit of the human NGF protein in mice.

[0105] (Example 3) Production of antibodies that neutralize the β-subunit (β-hNGF) of human NGF protein in immunized rabbits using peptides derived from β-hNGF. A peptide containing positions 122-135 of the β-subunit of the human NGF protein (derived from Sequence ID No. 1, see Table 4) was synthesized by solid-phase chemical synthesis using a standard method well known to those skilled in the art. This peptide was then coupled to the carrier protein CRM 197 using the coupling agent maleimide.

[0106] New Zealand rabbits (Agro-Bio, La Ferte-SFranceubin, France) were subcutaneously immunized with 25 μg of peptide emulsified in Montanide adjuvant (trademark) ISA 51 VG (n=5). The rabbits received three subcutaneous injections (day 0 (D0), day 28 (D28), and day 119 (D119)).

[0107] [Table 6]

[0108] The relative amount of anti-β-hNGF antibody is evaluated in rabbit serum by ELISA on days -10 (D-10 (pre-immunization)), 45 (D45), 80 (D80), 110 (D110), 136 (D136), 180 (D180), and 209 (D209).

[0109] In all immunized rabbits, we observed the production of antibodies that recognize the β-hNGF protein as early as day 45 (D45) (Figure 3). Antibodies that recognize the β-hNGF protein were present in the serum of all five immunized rabbits at all sample collection points. The peak level of antibodies that recognize the β-hNGF protein was observed at day 180 (D180), after the second booster immunization (day 119 (D119)).

[0110] The neutralizing ability of antibodies purified from the serum of each rabbit was evaluated using an ELISA-type test that inhibits the binding of β-hNGF protein to its hTrkA receptor. The produced antibodies neutralized more than 80% of 8 ng / mL of β-hNGF protein in all five immunized rabbits as early as day 45 (D45) (Figure 4). By day 209 (D209), the produced antibodies neutralized 100% of 8 ng / mL of β-hNGF protein in all five immunized rabbits (Figure 4). Conjugate peptides enable the induction of neutralizing antibodies against the β-subunit of human NGF protein in rabbits.

[0111] (Example 4) Production of antibodies that neutralize the β-subunit (β-hNGF) of human NGF protein in immunized rabbits using peptides derived from β-hNGF. A peptide containing positions 148-158 of the β-subunit of the human NGF protein (derived from Sequence ID No. 2, see Table 3) was synthesized by solid-phase chemical synthesis using a standard method well known to those skilled in the art, and then cyclized by forming a disulfide bridge between the two cysteine ​​units of the peptide. The peptide was then coupled to the carrier protein CRM 197 using a maleimide coupling agent.

[0112] New Zealand rabbits (Agro-Bio, La Ferte-St Aubin, France) were subcutaneously immunized with 25 μg of peptide emulsified in Montanide adjuvant (trademark) ISA 51 VG (n=5). The rabbits received three subcutaneous injections (day 0 (D0), day 28 (D28), and day 119 (D119)).

[0113] The relative amount of anti-β-hNGF antibody was evaluated in rabbit serum by ELISA on days -9 (D-9 (pre-immunization)), 46 (D46), 80 (D80), 110 (D110), 136 (D136), 180 (D180), and 209 (D209).

[0114] In all immunized rabbits, we observed the production of antibodies that recognize the β-hNGF protein as early as day 45 (D45) (Figure 5). Antibodies that recognize the β-hNGF protein were present in the serum of all five immunized rabbits at all sample collection points. The peak level of antibodies that recognize the β-hNGF protein was observed at day 180 (D180), after the second booster immunization (day 119 (D119)).

[0115] The neutralizing ability of antibodies purified from each rabbit's serum was evaluated using an ELISA-type test that inhibits the binding of the β-hNGF protein to its hTrkA receptor.

[0116] The produced antibodies neutralized the binding of the β-hNGF protein to its hTrkA receptor in 4 out of 5 immunized rabbits (Figure 6). Therefore, peptide conjugate induces neutralizing antibodies against the β-subunit of the human NGF protein in rabbits.

[0117] (Example 5) Evaluation of the analgesic effect of β-hNGF-derived peptides in a rat model of monosodium iodoacetate-induced osteoarthritis. Several experimental animal models of various species have been developed for human osteoarthritis, and these models have been used to study the preclinical efficacy of drug candidates that modify the disease and symptoms of osteoarthritis.

[0118] One such model involves chemical induction by intra-articular injection of monosodium iodoacetate (MIA) into the knee of rats. Intra-articular MIA disrupts the glycolysis pathway of chondrocytes by inhibiting glyceraldehyde-3-phosphate dehydrogenase. This leads to chondrocyte death and lesions throughout the joint cavity, causing symptoms of osteoarthritis, including joint pain, similar to those seen in human patients.

[0119] 8-10 week old Sprague Dolly rats (Cellvax, Institut Gustave Roussy, Hopital Paul Brousse, Villejuif, France) were randomly divided into three groups of 12 rats each (6 females and 6 males per group), designated as G1, G2, and G3. Rats in group G1 (vehicle group) were subcutaneously immunized with 0.1 mL of PBS emulsified with Montanide adjuvant® ISA 51 VG (n=12). Rats in group G2 were subcutaneously immunized with 0.1 mL of a peptide equivalent to 25 μg (SEQ ID NO: 23 (see Table 5)) emulsified with Montanide adjuvant® ISA 51 VG (n=12). G3 rats were subcutaneously immunized with 0.1 mL of a peptide equivalent to 25 μg emulsified in Montanide adjuvant (trademark) ISA 51 VG (SEQ ID NO: 22 (see Table 5)) (n=12). These peptides were derived from the sequences of two target peptide regions of rat β-NGF. The rats received three subcutaneous injections (day 0 (D0), day 28 (D28), and day 126 (D126)). A 1 mg injection of MIA was administered to the right knee 13 days after the third injection (day 139 (D139)).

[0120] [Table 7]

[0121] Evaluation of weight balance between the ipsilateral foot (MIA) and the contralateral foot.

[0122] The weight balance of each leg (left and right) of MIA-treated rats was analyzed using an incapacitometer. After the rats were allowed to acclimate in the incapacitometer for 5 minutes, both legs were fixed to a cushion and the weight balance was measured for 5 seconds. Three measurements were repeated in the same manner. The change in weight distribution between the two hind limbs was calculated as [(weight supported by the ipsilateral leg (MIA) / total weight supported by the ipsilateral and contralateral legs) × 100]. Percentage values ​​(%) were measured at 14 days (D14), 20 days (D20), 27 days (D27), 34 days (D34), and 42 days (D42) after MIA.

[0123] Weight balance between the ipsilateral leg (MIA) and the contralateral leg significantly improved at days 20 (D20), 34 (D34), and 42 (D42) in the group immunized with the two peptides (Figure 7), confirming analgesic effects linked to the anti-NGF response generated in each of the groups immunized with the two peptides tested, derived from SEQ ID NOs. 22 and 23 (the first and second sequences of rat NGF).

[0124] (Example 6) Production of antibodies in dogs that neutralize the β-subunit (β-hNGF) of the NGF protein by immunization with peptides derived from canine β-NGF. A peptide containing positions 122-135 of the β-subunit of the canine NGF protein (SEQ ID NO: 28, see Table 6) was synthesized by solid-phase chemical synthesis using a standard method well known to those skilled in the art. This peptide was then coupled to the carrier protein CRM 197 using the coupling agent maleimide.

[0125] Two dogs were subcutaneously immunized with a 25 μg equivalent of a peptide emulsified in Montanide adjuvant (trademark) ISA 51. The dogs received two subcutaneous injections on day 0 (D0) and day 28 (D28).

[0126] [Table 8]

[0127] When samples collected on day 2 (D-2) and day 42 (D42) were compared, ELISA revealed that both dogs produced antibodies recognizing human β-hNGF protein and canine peptides on day 42 (D42) (data not shown). The neutralizing ability of the antibodies was measured from canine serum. This evaluation was performed using an ELISA-type assay that inhibits the binding of human β-hNGF protein (added at 8 ng / mL) to its hTrkA receptor (attached to the bottom of the well). In this inhibition assay, human NGF protein can be used because the sequence of the targeted peptide (SEQ ID NO: 28) has only a single amino acid difference between human NGF and canine NGF.

[0128] The percentage of inhibition was calculated using a positive control antibody, whose neutralizing effect is known, as the baseline for 100% neutralization. In the experiment, serum from immunized dogs (collected on day 42 (D42)) neutralized approximately 50% of the β-hNGF protein, while serum from the same dogs before immunization (day -2 (D-2)) showed no neutralizing activity (Table 7). This result is interesting because, in addition to demonstrating that dogs immunized with canine NGF peptide can produce neutralizing antibodies, it also confirms that variant sequences can indeed induce antibodies that neutralize cytokines of other species (in this case, canine peptide induced antibodies that neutralize human NGF).

[0129] [Table 9]

Claims

1. - A first sequence consisting of at least eight consecutive amino acids selected from the sequence of amino acids 122 to 136 of the β-subunit of the NGF protein and up to 30 consecutive amino acids selected from the complete sequence of the β-subunit of the NGF protein, or a variant sequence having at least 75% identity with the first sequence; and / or - A second sequence consisting of at least eight consecutive amino acids selected from the sequence of amino acids 148 to 158 of the β-subunit of the NGF protein and up to 30 consecutive amino acids selected from the complete sequence of the β-subunit of the NGF protein, or a variant sequence having at least 75% identity with the second sequence. A polypeptide comprising or consisting thereof, However, polypeptides are different from the β-subunit of NGF protein, and do not consist of a continuous portion of more than 30 amino acids of the β-subunit of NGF protein, and However, the polypeptide comprising variant sequences of the first and second sequences, respectively, is capable of inducing an immune response directed towards the NGF protein.

2. The polypeptide according to claim 1, wherein the first sequence and the second sequence each consist of at least 12 consecutive amino acids selected from a sequence extending from amino acid positions 122 to 136 and a sequence extending from amino acid positions 148 to 158 of the β-subunit of an NGF protein, respectively.

3. The polypeptide according to claim 1 or 2, wherein the first sequence and the second sequence each consist at most of amino acids 122-136 and 148-158 of the β-subunit of the NGF protein.

4. The polypeptide according to any one of claims 1 to 3, wherein the β-subunit of the NGF protein is selected from the group consisting of the β-subunit of human NGF protein (hNGF), the β-subunit of mouse NGF protein, the β-subunit of rat NGF protein, the β-subunit of rabbit NGF protein, the β-subunit of ape NGF protein, the β-subunit of dog NGF protein, the β-subunit of cat NGF protein, the β-subunit of horse NGF protein, the β-subunit of pig NGF protein, the β-subunit of sheep NGF protein, and the β-subunit of dromedary camel NGF protein.

5. The polypeptide according to any one of claims 1 to 4, wherein the β-subunit of the NGF protein is a β-subunit of human NGF protein (hNGF) or canine NGF protein.

6. The first sequence and the second sequence are, - Sequence SSSHPIFHRGEFSVC (SEQ ID NO: 1) and sequence TATDIKGKEVM (SEQ ID NO: 2) or - Sequences SSSHPVFHRGEFSVC (SEQ ID NO: 28) and TATDIKGKEVM (SEQ ID NO: 27) A polypeptide according to any one of claims 1 to 5.

7. A polypeptide according to any one of claims 1 to 6, which is in a cyclic form.

8. A polypeptide according to any one of claims 1 to 7, linked to a carrier molecule.

9. A nucleic acid encoding a polypeptide according to any one of claims 1 to 8, or a complement thereof.

10. Depending on the case, together with at least one pharmaceutically acceptable carrier, the following: - At least one polypeptide according to any one of claims 1 to 8, or - At least one nucleic acid according to claim 9 A pharmaceutical composition containing as an active substance.

11. Use of a polypeptide according to any one of claims 1 to 8 for the preparation of antibodies, antibody fragments, or aptamers.

12. An anti-NGF antibody, antibody fragment, or aptamer specifically directed to the polypeptide described in any one of claims 1 to 8, However, the polypeptide is an antibody, antibody fragment, or aptamer that includes two or fewer amino acid residues in addition to the first sequence, the second sequence, or each of their variant sequences.

13. The antibody, antibody fragment, or aptamer according to claim 12 for use as a pharmaceutical.

14. A polypeptide according to any one of claims 1 to 8, a nucleic acid according to claim 9, or a pharmaceutical composition according to claim 10, for use in a method of inducing an immune response directed against an NGF protein in an individual.

15. A polypeptide according to any one of claims 1 to 8, a nucleic acid according to claim 9, a pharmaceutical composition according to claim 10, or an antibody, antibody fragment, or aptamer according to claim 13, for use in a method for preventing or treating a disease related to or caused by NGF protein in an individual.

16. Diseases related to or caused by NGF protein, - Allergic disorders, asthma disorders associated with respiratory hypersensitivity, typically characterized by episodes of cough and / or respiratory problems; - Multiple sclerosis, colitis, chronic inflammatory bowel disease, cystitis of the bladder, eczema, contact dermatitis, arthritis including chronic arthritis and rheumatoid arthritis, and psoriasis; - Osteoarthritis, lupus erythematosus, chronic obstructive pulmonary disease (COPD), lung cancer, hyperalgesia, chronic pain, and recurrent generalized or localized pain A polypeptide, nucleic acid, pharmaceutical composition, or antibody, antibody fragment or aptamer for use according to claim 15, selected from the group consisting of the following.

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