Active vaccination for the treatment of ngf-related disorders

Abstract

The present invention relates to veterinary compositions comprising an NGF antigen attached to a modified Cucumber Mosaic Virus (CMV) virus-like particle (VLP), in particular to a modified CMV VLP comprising a chimeric CMV polypeptide, for use in the treatment of NGF-related conditions in canids, in particular for use in the treatment of pain, such as canine pain associated with osteoarthritis (OA).

Description

[0001] This invention relates to a composition comprising an NGF antigen linked to a cucumber mosaic virus (CMV) virus-like particle (VLP), particularly linked to a modified CMV VLP comprising a chimeric CMV polypeptide, for use in the treatment of NGF-related diseases in canines, particularly for the treatment of pain (such as nociceptive pain, inflammation-related pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint diseases, and / or pain associated with osteoarthritis (OA)). The pain can be acute or chronic. Furthermore, this invention provides a highly immunogenic vaccine composition comprising an NGF antigen linked to a cucumber mosaic virus (CMV) virus-like particle (VLP), particularly linked to a modified CMV VLP comprising a chimeric CMV polypeptide, for active immunization of canines against NGF-related diseases, particularly pain.

[0002] Related technologies Nerve growth factor (NGF) was initially discovered to be a key factor in the development and maintenance of sensory and sympathetic neurons in the developing nervous system. In fact, NGF is essential for the survival of sensory and sympathetic neurons in the prenatal and early postnatal periods. However, in adults, the primary peripheral role of NGF shifts from providing nutritional support to sensory and sympathetic neurons to regulating the activity of nociceptive neurons. Preclinical and clinical studies over the past few decades have clearly demonstrated the important role of NGF in nociceptor sensitization in various acute and chronic pain states, including osteoarthritis (OA).

[0003] Osteoarthritis (OA)-related pain leads to decreased and altered mobility, which in turn causes localized and distal degeneration of the musculoskeletal system. Furthermore, the pathological processes of OA, such as joint capsule thickening and fibrosis, accelerate musculoskeletal changes that affect motor function. Finally, continuous nociceptive input to the CNS leads to changes in the somatosensory system and central sensitization, which also contribute to pain perception. In humans, the combined effects of pain, central sensitization, and activity impairment can negatively impact emotional states, exacerbating anxiety, depression, sleep disturbances, and cognitive impairment. Dogs with OA exhibit very similar consequences.

[0004] Osteoarthritis (OA) is a slowly progressive degenerative joint disease characterized by changes throughout the joint structure, including the articular cartilage, synovium, subchondral bone, and periarticular components, often leading to pain and loss of joint function. In canines (dogs), it is thought to primarily affect the hip, knee, and elbow joints, although a comprehensive prospective study on the prevalence of OA throughout the entire skeleton in canines has not been conducted. OA is often triggered early in life by developmental disorders such as hip dysplasia, but many other factors also play a role in its development, including diet, genetics, environment (including trauma), obesity, and age. OA is associated with clinical signs in a large proportion of the canine population, with an estimated 20% to 30% of dogs experiencing clinical impact. The disease is currently incurable and has negative consequences associated with pain, mobility impairment, and decreased quality of life.

[0005] Currently, pharmacological treatment of pain centers on nonsteroidal anti-inflammatory drugs (NSAIDs). These are used to relieve pain and promote functional improvement. Several NSAIDs are approved for use in dogs worldwide. Although NSAIDs are widely used and have significant benefits in many cases, they are not always effective as a single therapy. Furthermore, continuous treatment often produces side effects, including serious gastrointestinal and renal toxicity.

[0006] Recently, the use of anti-NGF therapeutic mAbs to alleviate pain in canine OA patients has been proposed (WO2022 / 076712). However, despite the benefits of mAb therapy, several challenges remain (e.g., high cost, the risk of anti-drug antibodies (ADA), allergic reactions due to the high protein content per dose, and the risk of local reactions). Furthermore, the duration of efficacy shown by mAbs is limited due to degradation and limited bioavailability.

[0007] Virus-like particles (VLPs) have been proposed and used in vaccine technology, particularly as immunological carriers to induce immune responses against conjugated antigens (Zeltins A, Mol Biotechnol (2013) 53:92-107; Jennings GT and Bachmann MF, Annu Rev Pharmacol Toxicol (2009) 49:303-26; Jennings GT and Bachmann MF, Biol Chem (2008) 389:521-536). Recently, the presentation of antigens, including autoantigens, on the surface of cucumber mosaic virus (CMV, family Bromoviridae, genus Cucumovirus) VLPs has been described using chemical linker conjugation technology (WO2016 / 062720) or fusion technology (WO2020 / 128037), and the induction of neutralizing antibody responses against certain targets.

[0008] However, effective treatment for canine pain, particularly active immunization against endogenous NGF using CMV VLPs, has not yet been demonstrated. Such effective treatment requires ensuring (i) adequate antibody levels and (ii) that the induced antibody response is controllable and irreversible, especially in the case of treatment targeting endogenous NGF proteins. Currently, there is no data indicating that NGF-related diseases, especially pain, such as pain associated with degenerative joint diseases like OA-related pain, can be controlled through active treatment using NGF-containing VLPs, particularly NGF-containing CMV VLPs.

[0009] Nevertheless, drug development still requires consideration of challenges and requirements, particularly in clinical trial testing, product registration, market launch, and commercial supply. Of particular note is the control of product characteristics such as stability, shelf life, solubility, manufacturability (including scalability), safety, potency, bioavailability, and other pharmacological properties, which are critical elements of the chemical, manufacturing, and control (CMC) process. These elements are essential for providing sufficient quantities of the product in a cost-effective manner to meet such end-user demands (Pham NG, Int J Pharm, 2020, 585:119523). Even under the various conditions required for efficient CMC processes, the stability of VLP-based vaccines is relevant. Another unintended event and problem that adversely affects product characteristics is the aggregation of biologics and vaccines (Roberts CJ, Current Opinion in Biotechnology, 2014, 30:211-217). Although aggregated vaccines may still elicit an immune response while retaining their natural structure, and therefore may still be suitable for some laboratory studies, their acceptance is lower for GMP-compliant products manufactured for clinical research and the market.

[0010] Therefore, given the current limitations in controlling NGF-related conditions (such as pain), particularly in controlling OA-related pain (whether acute or chronic), there is an unmet need to provide effective therapies to control emerging NGF-related conditions and to meet the requirements for final product registration to align with market release. Summary of the Invention

[0011] This invention provides compositions for treating NGF-related conditions, particularly for treating pain in canines. Specifically, the invention provides compositions for active immunization of canines against endogenous NGF proteins, producing high levels and durable neutralizing antibodies against these endogenous NGF proteins in such canines for active pain management. Specifically, the pain is selected from the group consisting of nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases, and / or osteoarthritis (OA)-related pain, preferably acute, chronic, and / or refractory-related pain.

[0012] Surprisingly, it has been found that the use of the compositions described herein, particularly in the form of active immunization against endogenous NGF proteins in canines, is advantageous compared to treatments using prior art. For example, the treatment provided herein induces a durable neutralizing anti-NGF antibody titer by temporarily overcoming natural immune tolerance. Furthermore, this long-acting neutralizing anti-NGF antibody eliminates the need for frequent administration of more expensive recombinant monoclonal antibodies (mAbs) and pharmaceutical agents.

[0013] Furthermore, the animal models used in this study not only demonstrated that administration of the cucumber virus (CMV) virus-like particle (VLP) composition containing NGF provided herein could generate high titers of durable neutralizing anti-NGF antibodies in vivo, but also provided evidence that active immunization with the NGF-containing CMV VLP composition unexpectedly alleviated NGF-related symptoms, particularly acute and chronic pain. Even more surprisingly, treatment with the NGF-containing CMV VLP composition was superior in pain management compared to antibody therapy and NSAID therapy. Moreover, even though the treatment targeted an autoprotein (NGF protein), the immune response induced by the autoprotein did not cause any side effects in canines, while effectively managing NGF-related diseases, especially pain.

[0014] Therefore, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0015] Specifically, NGF-related conditions are pain, preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint diseases, and / or pain associated with osteoarthritis (OA). Data show that such treatments can alleviate acute, chronic, and refractory pain.

[0016] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related symptom is pain.

[0017] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, preferably wherein such pain is OA-related pain.

[0018] The animal data provided herein indicate that the CMV VLP composition containing NGF can treat, particularly relieve, acute, chronic, and refractory pain. According to another aspect, acute pain is acute pain associated with degenerative joint disease, preferably acute OA-related pain. According to another aspect, chronic pain is chronic pain associated with degenerative joint disease, preferably chronic OA-related pain. According to another aspect, chronic pain is refractory chronic pain associated with degenerative joint disease, preferably refractory chronic OA-related pain.

[0019] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is pain associated with degenerative joint disease, preferably selected from acute pain associated with degenerative joint disease, chronic pain associated with degenerative joint disease, and refractory chronic pain associated with degenerative joint disease.

[0020] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is OA-related pain, preferably selected from acute OA-related pain, chronic OA-related pain and refractory chronic OA-related pain.

[0021] The CMV VLP composition containing NGF enhances neutralizing anti-NGF antibodies upon repeated or multiple administrations without inducing any immune tolerance, thus avoiding any side effects or attenuation of the adjuvant's effect on endogenous NGF proteins. The lack of immune tolerance against CMV VLP and the linked NGF antigen after repeated administration further improves therapeutic efficacy in managing NGF-related conditions, particularly in pain management.

[0022] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in one or more doses.

[0023] In cases of repeated administration, the time interval between the first and second doses is preferably at least 7 days, more preferably at least 14 days, and even more preferably 7 to 21 days, with 14 to 21 days being particularly advantageous. Further booster administrations are typically provided at longer intervals. For example, if a third dose is provided, the selected time interval between the second and third doses is generally two to six months, such as three months. For any further booster administrations, the selected time interval is at least three months, preferably three to six months, and most preferably four to six months.

[0024] To relieve acute pain, one or two doses of a CMV VLP composition containing NGF are typically administered. If two doses are administered, a time interval of one to three weeks, preferably two to three weeks, is usually chosen to obtain the maximum therapeutic effect.

[0025] To relieve chronic pain, multiple applications can enhance the therapeutic effect. If three to four doses are administered, the interval between the first and second doses is typically one to three weeks, preferably two to three weeks, the interval between the second and third doses is two to six months, for example two to three months, and the interval between any fourth dose and the third dose is selected to be three to six months, preferably four to six months.

[0026] Generally, CMV VLP compositions containing NGF are administered systemically, preferably subcutaneously, intramuscularly, or transdermally. In the selected animal models, subcutaneous administration was chosen.

[0027] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered systemically, preferably subcutaneously, intramuscularly or transdermally, preferably subcutaneously.

[0028] There are different ways in which CMV VLPs and NGF antigens can be linked. NGF antigens can be linked to CMV VLPs via attachment sites, through peptide bonds, preferably through gene fusion. For example, the nucleotide sequence encoding the NGF antigen can be cloned within the inner frame of the nucleotide sequence encoding the CMV VLP. Alternatively, the NGF antigen can be linked to the CMV VLP via chemical coupling between attachment sites, for example, through at least one non-peptide bond.

[0029] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are fused together via at least one covalent peptide bond through at least one first attachment site and at least one second attachment site.

[0030] According to another aspect, the present invention provides a composition for a method of treating NGF-related conditions (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected via at least one covalent nonpeptide bond through at least one first attachment site and at least one second attachment site.

[0031] CMV VLPs are typically large molecular assemblies composed of viral capsid proteins, typically containing 180 protein subunits per VLP. Typically and preferably, the interaction of these subunits leads to the formation of VLPs with inherently repeating organization, thereby allowing the presentation of multiple copies of the NGF antigen. Such capsid proteins are, for example, capsid proteins encoded by the amino acid sequence SEQ ID NO: 39 or having at least 75% sequence identity with SEQ ID NO: 39.

[0032] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A CMV VLP, wherein the CMV VLP contains at least one first attachment site, and wherein the CMV VLP contains at least one CMV polypeptide, wherein the CMV polypeptide contains a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO: 39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the NGF antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site. Preferably, the NGF antigen and the CMV VLP are connected via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site.

[0033] The immunogenicity of the NGF-containing CMV VLP composition described herein can be further enhanced by adding a helper T (Th) cell epitope. Preferably, the helper T cell epitope is a helper T cell epitope derived from tetanus toxin or a PADRE sequence. Such helper T cell epitopes preferably contain or consist of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Such helper T cell epitopes can be introduced, preferably into CMV VLPs, particularly into CMV peptides, and preferably into the coat protein sequence of CMV VLPs. According to a preferred specific example of the CMV VLP containing the helper T cell epitope, the helper T cell epitope, preferably a helper T cell epitope derived from tetanus toxin or a PADRE sequence, replaces the N-terminal region of the CMV peptide. Preferably, a helper T cell epitope, preferably derived from tetanus toxin or a PADRE sequence, replaces the N-terminal region of the CMV polypeptide, and preferably, the N-terminal region of the CMV polypeptide corresponds to amino acid residues 2-12 of SEQ ID NO: 39.

[0034] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen comprises at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (c) Helper T cell epitopes; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0035] According to another aspect, the present invention provides a composition for a method of treating NGF-related conditions (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen comprises at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (c) A helper T cell epitope, wherein the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, preferably wherein the helper T cell epitope comprises or is composed of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0036] According to another aspect, the present invention provides a composition for a method of treating NGF-related conditions (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen comprises at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (c) A helper T cell epitope, preferably derived from tetanus toxin or a PADRE sequence, preferably comprising or consisting of SEQ ID NO:41 or SEQ ID NO:42, replacing the N-terminal region of the CMV polypeptide, preferably corresponding to the N-terminal region of amino acid residues 2-12 of SEQ ID NO:39; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0037] In another aspect, the present invention provides a composition for a method of treating NGF-related conditions (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises a CMV polypeptide, preferably composed of a CMV polypeptide, wherein the CMV polypeptide comprises a CMV coat protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO: 39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the NGF antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked through at least one first attachment site and at least one second attachment site. Preferably, the NGF antigen and the modified CMV VLP are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. In a preferred embodiment, the modified CMV polypeptide comprises, and preferably consists of, the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 48.

[0038] In addition to or as an alternative to immunogenic properties, the physicochemical properties of CMV VLPs can be further improved by inserting segments of consecutive negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid. Such CMV VLPs exhibit improved stability at high temperatures and higher ionic strengths. Furthermore, when conjugated with NGF antigens, these CMV VLP-NGF antigen conjugates do not form aggregates and remain stable in solution after NGF antigen conjugation, whereas prior art CMV VLPs form a certain amount of aggregates and precipitates. These improved (non-aggregating) CMVVLP-NGF antigen conjugates are highly desirable for drug development and product registration.

[0039] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising a continuous segment of negatively charged amino acids, preferably composed therefrom, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, wherein the polypeptide is inserted into the CMV polypeptide of (i). (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site.

[0040] Preferably, the segment of the continuous negatively charged amino acid, independently selected from aspartic acid or glutamic acid, is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO: 39.

[0041] This continuous segment of negatively charged amino acids typically has a length of 3 to 10 amino acids, and, on the other hand, may consist of only glutamic acid.

[0042] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related symptoms (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising a continuous segment of negatively charged amino acids (preferably 3 to 10 amino acid residues), preferably composed thereof, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO: 39. (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected via at least one first attachment site and at least one second attachment site, preferably via at least one covalent non-peptide bond. Preferably, the chimeric CMV polypeptide comprises, and preferably consists of, the amino acid sequence of SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 12.

[0043] According to another aspect, the continuous negatively charged amino acid segment further includes a first amino acid linker and a second amino acid linker, wherein the first amino acid linker is located at the N-terminus of the continuous negatively charged amino acid segment, and the second amino acid linker is located at the C-terminus of the continuous negatively charged amino acid segment, and wherein the first amino acid linker and the second amino acid linker are independently selected from the group consisting of: i) Polyglycine linkers (G-linkers) having an amino acid sequence (Gly) of length n = 2 to 10. n ; ii) A glycine-serine linker (GS-linker) comprising at least one glycine and at least one serine, wherein preferably, the GS-linker has the amino acid sequence (GS)r(GsS)t(GS)u, where r = 0 or 1, s = 1-5, t = 1-5 and u = 0 or 1; and iii) An amino acid linker (GS*-linker) comprising at least one Gly, at least one Ser and at least one amino acid selected from Thr, Ala, Lys and Cys.

[0044] According to another aspect, the polypeptide containing the continuous segment of negatively charged amino acids is composed of SEQ ID NO: 49, SEQ ID NO: 50 or SEQ ID NO: 51.

[0045] According to another aspect, the present invention provides a composition for a method of treating NGF-related conditions (preferably pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particles (VLPs), wherein the CMV VLP contains at least one first attachment site and further contains at least one helper T cell epitope, and wherein such CMV VLP contains, preferably constitutes, the amino acid sequence of SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 12; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected by at least one first attachment site and at least one second attachment site, preferably by at least one non-peptide bond.

[0046] The NGF antigen used in any NGF-containing CMV VLP composition in the method for treating NGF-related diseases in canines as described herein may be any NGF antigen that exhibits the desired technical effect in canines. Preferably, the NGF antigen is canine NGF antigen (cNGF). According to another aspect, the NGF antigen used in the NGF-containing CMV VLP composition in the method for treating NGF-related diseases in canines according to the invention comprises or preferably consists of the following: an amino acid sequence selected from any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58, or an amino acid sequence having at least 90%, preferably at least 95%, sequence identity with any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58.

[0047] According to another aspect, the NGF antigen used in the NGF-containing CMV VLP composition in the method for treating NGF-related diseases in canines according to the present invention comprises or preferably consists of the following: an amino acid sequence selected from any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33 and SEQ ID NO:55, or an amino acid sequence having at least 90%, preferably at least 95%, sequence identity with any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33 and SEQ ID NO:55. Brief description of the attached diagram Figure 1 Description of the pET-CMVB2-Ntt-E8* plasmid map containing a single restriction enzyme site.

[0049] Figure 2ASDS-PAGE gel analysis of purified VLPs expressing CMV-Ntt830-E8*. M - Protein size marker PageRuler (Thermo Fisher Scientific, #26620); S - Soluble protein / pET-CMVB2-Ntt-E8* in E. coli C2566 cell extract; P - Insoluble protein in cell extract; 1 - Insoluble protein after sucrose gradient (bottom of tube); 2 - 6 - Sucrose gradient fraction (60% from bottom to 0% from top of tube). Asterisks (*) in the figure indicate the relative position of the corresponding CMV-Ntt830-E8* chimeric CMV peptide in the SDS / PAGE gel.

[0050] Figure 2B Electron micrograph of purified CMV-Ntt830-E8* VLP. Horizontal bars correspond to 500 nm.

[0051] Figure 3 Description of the pET-CMVB2-Ntt-E4 plasmid map containing a single restriction enzyme site.

[0052] Figure 4 Description of the pET-CMVB2-Ntt-E8 plasmid map containing a single restriction enzyme site.

[0053] Figure 5 Description of the pET-CMVB2-Ntt-E12 plasmid map containing a single restriction enzyme site.

[0054] Figure 6 SDS-PAGE (left) and agarose gel (right) analysis of purified VLPs expressing CMV-Ntt830-E4. M1 - Protein size marker PageRuler (Thermo Fisher Scientific, #26620); M2 - DNA size marker (Thermo Fisher Scientific, #SM0311); T - Total protein in E. coli C2566 cells after 18 hours of culture at 20°C; S - Soluble protein in cell extracts before sucrose gradient (20-60%) after cell lysis; P - Insoluble protein; 1-6 - Sucrose gradient fractions (60% at the bottom of the tube to 0% at the top). The asterisks (*) in the figure indicate the relative positions of the corresponding CMV-Ntt830-E4 chimeric CMV peptides in the SDS / PAGE gel and the relative positions of typical VLP signals in the agarose gel.

[0055] Figure 7SDS-PAGE (left) and agarose gel (right) analysis of purified VLPs expressing CMV-Ntt830-E8. M1 - PageRuler protein size marker (Thermo Fisher Scientific, #26620); M2 - DNA size marker (Thermo Fisher Scientific, #SM0311); T - Total protein in E. coli C2566 cells after 18 hours of culture at 20°C; S - Soluble protein in cell extracts before sucrose gradient (20-60%) after cell lysis; P - Insoluble protein; 1-6 - Sucrose gradient fractions (60% at the bottom of the tube to 0% at the top). The asterisks (*) in the figure indicate the relative positions of the corresponding CMV-Ntt830-E8 chimeric CMV peptides in the SDS / PAGE gel and the relative positions of typical VLP signals in the agarose gel.

[0056] Figure 8 SDS-PAGE (left) and agarose gel (right) analyses of purified VLPs expressing CMV-Ntt830-E12. M1 - Protein size marker PageRuler (Thermo Fisher Scientific, #26620); M2 - DNA size marker (Thermo Fisher Scientific, #SM0311); T - Total protein in *E. coli* C2566 cells after 18 hours of incubation at 20°C; S - Soluble protein in cell extracts before sucrose gradient (20-60%) after cell lysis; P - Insoluble protein; 1-6 - Sucrose gradient fractions (60% at the bottom to 0% at the top). The asterisks (*) in the figure indicate the relative positions of the corresponding CMV-Ntt830-E12 chimeric CMV peptides in the SDS / PAGE gel. No clear bands corresponding to intact VLPs were observed in the agarose gel.

[0057] Figure 9 Electron micrograph of purified CMV-Ntt830-E4 VLP. Horizontal bars correspond to 200 nm.

[0058] Figure 10 Electron micrograph of purified CMV-Ntt830-E8 VLP. Horizontal bars correspond to 200 nm.

[0059] Figure 11Comparison of the thermal stability of CMV-Ntt830 VLP and CMV-Ntt830-E4 VLP. The structural changes of CMV-Ntt830 VLP and CMV-Ntt830-E4 VLP in the presence of Sypro-Orange dye were monitored using a DNA melting temperature assay program and a real-time PCR system. Curve 1 represents CMV-Ntt830-E4 VLP, curve 2 represents CMV-Ntt830 VLP, and curve 3 represents the buffer control (5 mM sodium phosphate, 2 mM EDTA, pH 7.5). The corresponding melting temperatures of 57℃ and 51℃ are indicated by arrows.

[0060] Figure 12 Stability of CMV-Ntt830 VLP and CMV-Ntt830-E4 VLP in solution with different concentrations of NaCl. 0.5 mg / ml CMV-Ntt830 VLP and CMV-Ntt830-E4 VLP samples were incubated at room temperature for up to 2 hours in 5 mM sodium phosphate, 2 mM EDTA (pH 7.5), and different concentrations of NaCl (the molar concentration of NaCl in each sample is indicated at the bottom of the gel). Samples were analyzed by natural agarose gel electrophoresis and ethidium bromide staining. Figures A and B show the NAGE analysis of CMV-Ntt830 VLP and CMV-Ntt830-E4 VLP samples, respectively. M shows the lane loaded with the GeneRuler 1kb DNA gradient (SM0311, TFS). Black arrows indicate the location of the loaded wells within the gel and the positions of the VLPs in each well and within the gel. The presence of CMV-Ntt830 VLPs (Patch A) in the loading wells after electrophoresis is due to the formation of VLP aggregates that are too large to enter the gel. Unaggregated, intact VLPs migrate into the gel.

[0061] Figure 13Analysis of CMV-Ntt830 VLPs after anion exchange chromatography. 5 ml of 5 mM sodium borate buffer (pH 9.0) containing 1 mg / ml CMVtt-VLP was loaded onto a 1.0 ml Macro-Prep DEAE Bio-Rad anion exchange filter cartridge equilibrated with 5 mM sodium borate buffer and eluted stepwise in the presence of increasing NaCl concentrations (0.1, 0.2, 0.3, 0.4, 0.5, 0.8, 1.0, and 2.0 M). Fractions were collected and analyzed by nanodrop 260 nm (for protein concentration) and native agarose gel electrophoresis. Figure A shows the NaCl concentration and absorbance at 260 nm for each fraction (1-25). Figure B shows the NAGE analysis (ethidium bromide staining) of the major fraction containing the highest protein concentration. M shows the lane loaded with a GeneRuler 1kb DNA gradient (SM0311, TFS). The black arrows indicate the location of the loading wells within the gel, as well as the location of VLPs within each well and the gel. The presence of CMV-Ntt830 VLPs in the loading wells after electrophoresis is due to the formation of VLP aggregates that are too large to enter the gel. Unaggregated, intact VLPs migrate into the gel.

[0062] Figure 14Analysis of CMV-Ntt830-E4 VLPs after anion exchange chromatography. Biomass from *E. coli* cells expressing CMV-Ntt830-E4 VLPs was resuspended in 50 mM citrate, 5 mM borate buffer at pH 9.0, and cells were lysed using an LM-20 microfluidic bed. The soluble fraction was clarified by centrifugation and loaded onto a 60 ml Fracto-DEAE (XK26 / 20) column. The bound VLPs were eluted using a continuous gradient of elution buffer containing 50 mM citrate, 5 mM borate, and 1 M NaCl. Figure A shows the protein elution and NaCl concentration gradients measured at A260 nm (mAU) and conductivity (mS / cm), respectively. The X-axis shows the elution volume and fraction number (4-11). Fractions collected from the Fracto-DEAE column were analyzed by NAGE (plot B) and SDS-PAGE (plot C). In panel B, M indicates the lane loaded with the GeneRuler 1kb DNA gradient (SM0311, TFS), L is the E. coli lysate sample before loading onto the Fracto DEAE, and FT represents the flow-through collected from 0 to 150 ml, with 4-10 representing the fraction numbers collected during elution. The black arrows, from top to bottom, indicate the location of the loading well, the position of the intact CMV-Ntt830-E4 VLP within the gel, and the location of contaminating nucleic acids from the clarified bacterial lysate. In panel C, FT represents the flow-through collected from 0 to 150 ml, with 4-10 representing the fraction numbers. The black arrows show the location of the CMV-Ntt830-E4 capsid protein stained with Coomassie blue.

[0063] Figure 15A Purification and purity of recombinant mature canine NGF. SDS-PAGE analysis of the NGF purification process. M - Label, with molecular weight of bands shown in kDa; A - Total cell lysate after expression; B - Mixed fraction containing pro-NGF after refolding and partial purification; C - Mature NGF after trypsin digestion and final purification. Arrows indicate pro-NGF in lanes A and B and mature NGF in lane C.

[0064] Figure 15B PC12 cells were cultured for 5 days with recombinant human mature NGF (R&D systems) produced from mouse myeloma cells (black squares) or with canine mature NGF produced from *E. coli* as described herein (gray circles). Cells were cultured at 100, 50, 25, 12.5, and 6.25 ng / ml of recombinant NGF, and the percentage of cells exhibiting defined neurite overgrowth was determined.

[0065] Figure 16A SDS-PAGE analysis of recombinant mature canine NGF (cNGF) conjugated with CMV-Ntt830 and CMV-Ntt830-E8* VLP in Seq ID NO:31.

[0066] M - PageRuler™ Plus pre-stained protein gradient, 10 to 250 kDa (Thermo Fisher Scientific, # 26620) protein size markers; 1 - Corresponding purified CMV-Ntt830 and CMV-Ntt830-E8* VLPs; 2 - CMV VLPs derivatized with 5 × SMPH and then SMPH removed; 3 - CMV VLPs conjugated with an equimolar amount of cNGF; 4 - A mixture of CMV-Ntt830-E8* and cNGF without SMPH derivatization; 5 - Purified cNGF. An asterisk indicates the location of observable CMV VLP-NGF conjugate bands.

[0067] Figure 16B SDS-PAGE analysis of recombinant mature canine NGF (cNGF) of SEQ ID NO:31 conjugated with CMV-Ntt830-E4 and CMV-Ntt830-E8 VLP.

[0068] M - PageRuler™ Plus pre-stained protein gradient, 10 to 250 kDa (Thermo Fisher Scientific, # 26620) protein size markers; 1 - Corresponding purified CMV-Ntt830-E4 and CMV-Ntt830-E8 VLPs; 2 - CMV VLPs derivatized with 5 × SMPH and then SMPH removed; 3 - CMV VLPs conjugated with an equimolar amount of cNGF; 4 - Mixed samples of CMV-Ntt830-E4 or CMV-Ntt830-E8 with cNGF without SMPH derivatization; 5 - Purified cNGF. An asterisk indicates the location of observable CMV VLP-cNGF conjugate bands.

[0069] Figure 16C Dynamic light scattering analysis of cNGF-CMV-Ntt830 VLP. EM analysis was not possible due to vaccine precipitation.

[0070] Figure 16D Dynamic light scattering analysis of cNGF-CMV-Ntt830-E4 VLP containing the cNGF antigen of SEQ ID NO:31.

[0071] Figure 16E Dynamic light scattering analysis of cNGF-CMV-Ntt830-E4 VLP containing cNGF antigen of SEQ ID NO:33.

[0072] Figure 16F Dynamic light scattering analysis of cNGF-CMV-Ntt830-E8* VLP.

[0073] Figure 16G Electron microscopy of cNGF-CMV-Ntt830-E4 VLP.

[0074] Figure 16H Electron microscopy of cNGF-CMV-Ntt830-E8* VLP.

[0075] Figure 17A : Assess anti-NGF IgG antibodies in the serum of dogs immunized with cNGF-CMV-Ntt830-E8* VLP. Anti-NGF IgG titers in Group 1 dogs that received the vaccine but not the adjuvant. Arrows indicate the vaccination administered on days 0, 21, and 42.

[0076] Figure 17B : To evaluate anti-NGF IgG antibodies in the serum of dogs immunized with cNGF-CMV-Ntt830-E8* VLP. Anti-NGF IgG titers in 3 dogs in Group 2 who received the vaccine and adjuvant QuilA®. Arrows indicate the vaccination administered on days 0, 21, and 42.

[0077] Figure 17C : Assess anti-CMV IgG titers in the serum of dogs immunized with cNGF-CMV-Ntt830-E8*VLP. Anti-CMV IgG titers in Group 1 dogs that received the vaccine but not the adjuvant. Arrows indicate the vaccination administered on days 0, 21, and 42.

[0078] Figure 17D : Assess anti-CMV IgG titers in the serum of dogs immunized with cNGF-CMV-Ntt830-E8* VLP. Anti-CMV IgG titers in Group 2 dogs that received the vaccine and adjuvant QuilA®. Arrows indicate the vaccination administered on days 0, 21, and 42.

[0079] Figure 17ETo evaluate anti-NGF IgG antibodies in the serum of dogs immunized with cNGF-CMV-Ntt830-E4 VLP in the absence of adjuvant. Five dogs were administered cNGF-CMV-Ntt830-E4 VLP on days 0 and 21. NGF-specific antibodies in sera collected on days 0, 21, 42, 71, and 91 were assessed by ELISA.

[0080] Figure 17F To evaluate anti-NGF IgG antibodies in the serum of dogs immunized with cNGF-CMV-Ntt830-E4 VLP in the presence of aluminum hydroxide. Five dogs were administered cNGF-CMV-Ntt830-E4 VLP with aluminum hydroxide on days 0 and 21. NGF-specific antibodies were measured by ELISA on days 0, 21, 42, 71, and 91.

[0081] Figure 18A Dogs were immunized with cNGF-CMV-Ntt830-E8* VLP to induce NGF neutralizing antibodies. Dogs (n=3 per group) were immunized with 250 μg of cNGF-CMV-Ntt830-E8* VLP on days 0, 21, and 42, with or without the adjuvant QuilA. Serum was collected and the presence of neutralizing antibodies was tested using a TF-1-based NGF bioactivity assay. A titration curve for one dog was used to determine the neutralizing capacity and 50% neutralizing titer (NT50) of the canine serum. 5 ng / mL of mature human NGF was pre-incubated with progressively increasing concentrations of purified IgG collected from serum on specified dates following the first administration of the vaccine. The NT50 value, i.e., the IgG concentration that results in 50% inhibition of cell proliferation, was determined using a 4PL sigmoid curve fitting model.

[0082] Figure 18BDogs were immunized with cNGF-CMV-Ntt830-E8* VLP on days 0, 21, and 42, with or without the adjuvant QuilA (3 dogs per group). Serum was collected and the presence of neutralizing antibodies was tested using a TF-1-based NGF bioactivity assay. Total IgG was purified from the dog serum. The ability of 20 μg / mL purified total IgG to neutralize 5 ng / mL mature human NGF was assessed using a bioanalyte. Bars represent the group mean versus standard deviation, and the sign represents the individual dog (mean of two replicate analyses). Two-factor analysis of variance and Tukey's multiple comparisons test were performed using GraphPad Prism to compare group means. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

[0083] Figure 18C Vaccination with cNGF-CMV-Ntt830-E8* VLP induced the production of mature NGF neutralizing antibodies in dogs. Dogs were immunized with 250 μg of cNGF-CMV-Ntt830-E8* VLP on days 0, 21, and 42, with or without the adjuvant QuilA (n=3 per group). Serum was collected and the presence of neutralizing antibodies was tested using a TF-1-based NGF bioactivity assay. NT50 values ​​were plotted against OD50 values ​​of anti-NGF IgG serum titers. Total IgG purified from serum with higher concentrations of NGF-specific antibodies inhibited NGF-mediated TF-1 cell proliferation more effectively than total IgG purified from serum of dogs with lower anti-NGF titers. Symbols represent individual dogs and sampling time points. Different symbols were assigned to different dogs. Closed symbols represent animals vaccinated in the presence of adjuvants, while open symbols represent animals vaccinated in the absence of adjuvants.

[0084] Figure 18D Vaccination with cNGF-CMV-Ntt830-E4 VLP induced NGF neutralizing antibodies in dogs. On days 0 and 21, cNGF-CMV-Ntt830-E4 VLP was administered to five dogs along with aluminum hydroxide. Serum was collected on day 42, and a TF-1-based NGF bioactivity assay was used to test for the presence of neutralizing antibodies. Bars represent the cohort mean and standard deviation, and the symbols represent individual dogs. The dashed line indicates the detection limit of the assay.

[0085] Figure 19AKaolin model of acute inflammatory pain: Dogs were vaccinated with cNGF-CMV-Ntt830-E4 VLP (square) compared to those treated with the prior art monoclonal antibody betinvetmab (diamond) and placebo (triangle). The cNGF-specific IgG titer (GMT) in the study group is plotted as EC50 values. Error bars represent the 95% CI of GMT. Analytical results below LoD were set at 50, corresponding to 0.5 times the lowest dilution used in the analysis (i.e., 1:100) (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0086] Figure 19B Kaolin model of acute inflammatory pain: Dogs were vaccinated with cNGF-CMV-Ntt830-E4 VLP (square) to be treated with the previously used monoclonal antibody betinvelumab (triangle) and placebo (circle)—the cNGF-specific IgG titer (GMT) in the study groups is plotted as a standardized equivalent concentration. Error bars represent the 95% CI of GMT. Titers above LoD but below LoQ were set at 100. A peak antibody titer in serum after betinvelumab administration has been previously reported to be 6.1 μg / mL (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0087] Figure 20A Kaolin model of acute inflammatory pain: Dogs were vaccinated with cNGF-CMV-Ntt830-E4 VLP (square) to be treated with the prior art monoclonal antibody betinvelumab (triangle) and placebo (circle) – NGF neutralizing titers were determined by a TrkA-based NGF binding ELISA. The NGF neutralization GMT in the study group is plotted as the EC50 value. Error bars represent the 95% CI of GMT. Analytical results below LoD were set at 50, corresponding to 0.5 times the lowest dilution used in the analysis (i.e., 1:100) (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0088] Figure 20BKaolin model of acute inflammatory pain: Vaccination of dogs using cNGF-CMV-Ntt830-E4 VLP (square) was compared with treatment using the previously developed monoclonal antibody betinvelumab (triangle) and placebo (circle) – NGF neutralizing titers were determined by a TrkA-based NGF binding ELISA. The NGF neutralizing GMT in the study group is plotted as the standard equivalent concentration. Error bars represent the 95% CI of GMT. Titers above LoD but below LoQ were set at 100. A peak antibody titer in serum after betinvelumab administration was previously reported to be 6.1 μg / mL (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0089] Figure 21A Kaolin model of acute inflammatory pain: Vaccination of dogs with cNGF-CMV-Ntt830-E4 VLP (square) was compared with treatment with the previously used monoclonal antibody betinvelumab (triangle) and placebo (circle) – NGF neutralization GMT was determined by NGF bioassay based on TF-1 cells. NGF neutralization GMT in the study group is plotted as EC50 value. Error bars represent the 95% CI of GMT. Analytical results below LoD were set at 50, corresponding to 0.5 times the lowest dilution factor used in the analysis (i.e., 1:100) (dashed line). GMT is equivalent to geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0090] Figure 21B Kaolin model of acute inflammatory pain: Vaccination of dogs with cNGF-CMV-Ntt830-E4 VLP (square) was compared with treatment with the previously used monoclonal antibody betinvelumab (triangle) and placebo (circle) – NGF neutralization GMT was determined by NGF bioassay based on TF-1 cells. NGF neutralization GMT in the study group is plotted as the standard equivalent concentration. Error bars represent the 95% CI of GMT. Titers above LoD but below LoQ were set at 100. A peak antibody titer in serum after betinvelumab administration was previously reported to be 6.1 μg / mL (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0091] Figure 22Surgical model of acute pain: Seven dogs in four groups were treated with cNGF-CMV-Ntt830-E4 VLP vaccine, monoclonal betinvelumab (Librela™), meloxicam (Inflacam™), and placebo (water for injection), respectively. Treatment was administered 21 and 42 days before, and 7, 21, and 42 days before, initiating the inflammatory / acute pain model, which was induced by injecting a kaolin chemical into the right hind paw of all dogs. Lameness scores were assessed using a validated numerical rating scale based on the typical course of the disease in each dog. The numerical rating scale used to assess lameness in inflamed paws was based on examiner perception and the scores were determined as follows: (0) no lameness; (1) barely detectable lameness for most of the observation period; (2) mild lameness with maximum weight-bearing capacity; (3) moderate lameness with minimum weight-bearing capacity; (4) severe lameness where the animal uses its paw (initiating walking movements and / or lightly touching the ground) but does not bear weight; (5) the most severe lameness where the animal refuses to move and / or avoids any contact between the inflamed paw and the ground. Lameness scores were measured in all dogs over a two-week period following the kaolin attack.

[0092] Figure 23 During the 14-day period following kaolin challenge, the area under the curve (AUC) of lameness scores was calculated for dogs treated with cNGF-CMV-Ntt830-E4 VLP vaccine, monoclonal betinvelumab (Librela™), meloxicam (Inflacam™), and placebo (water for injection).

[0093] Figure 24A Surgical experimental model of chronic pain: Dogs vaccinated with cNGF-CMV-Ntt830-E4 VLP (Group A) - The cNGF-specific IgG titer (GMT) in the study group is plotted as the EC50 value. Error bars represent the 95% CI of GMT. Analytical results below LoD are set at 50, corresponding to 0.5 times the lowest dilution used in the analysis (i.e., 1:100) (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0094] Figure 24B Surgical experimental model of chronic pain: Dogs vaccinated with cNGF-CMV-Ntt830-E4 VLP (Group B) - The cNGF-specific IgG titer (GMT) in the study group is illustrated as the standard equivalent concentration. Error bars represent the 95% CI of GMT. Titers above LoD but below LoQ are set at 100. Previously reported peak antibody titers in serum after betinvelumab administration were 6.1 μg / mL (dashed line). GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0095] Figure 25A Geometric mean NGF neutralization titer determined by TrkA-based NGF binding ELISA. The NGF neutralization GMT for the study group is plotted as the EC50 value. Error bars represent the 95% CI of the GMT. Analytical results below LoD are set at 50, corresponding to 0.5 times the lowest dilution used in the analysis (i.e., 1:100) (dashed line). Titers above LoD but below LoQ are set at 100. GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0096] Figure 25B Geometric mean NGF neutralization titer as determined by TrkA-based NGF binding ELISA. The NGF neutralization GMT in the study group is plotted as the standard equivalent concentration. Error bars represent the 95% CI of GMT. A peak antibody titer in serum after betinvelumab administration was reported to be 6.1 μg / mL (dashed line). Geometric mean titer of GMT; LoD = limit of detection; LoQ = limit of quantitation.

[0097] Figure 26A NGF neutralization GMT was determined by NGF bioanalysis based on TF-1 cells. The NGF neutralization GMT for the study group is plotted as the EC50 value. Error bars represent the 95% CI of GMT. Analytical results below LoD were set at 50, corresponding to 0.5 times the lowest dilution used in the analysis (i.e., 1:100) (dashed line). Titers above LoD but below LoQ were set at 100. GMT is equivalent to the geometric mean titer; LoD = limit of detection; LoQ = limit of quantitation.

[0098] Figure 26B NGF-neutralizing GMT was determined by NGF bioanalysis based on TF-1 cells. The NGF-neutralizing GMT in the study group is plotted as the standard equivalent concentration. Error bars represent the 95% CI of GMT. A peak antibody titer in serum after administration of betinvelumab was reported to be 6.1 μg / mL (dashed line). Geometric mean titer of GMT; LoD = limit of detection; LoQ = limit of quantitation.

[0099] Figure 27A Surgical model of chronic pain: Seven dogs in two groups were treated with cNGF-CMV-Ntt830-E4 VLP vaccine and placebo (water for injection), respectively. Treatment was performed approximately 21 days and 42 days before the implementation of the experimental model of chronic pain caused by right-sided stiff joint instability due to surgery.

[0100] Localized pain in the treated joints was assessed by palpation and scored according to a predefined pain palpation grading scale (range 0 to 4) derived from the Modified Glasgow Composite Pain Scale. Palpation response (touch pain) scores were measured at predetermined time points from study day 84 (SD84) to SD217 in all dogs.

[0101] Figure 27B Surgical experimental model of chronic pain: Dogs were vaccinated with cNGF-CMV-Ntt830-E4 VLP (Group B) - The cNGF-specific IgG titer (GMT) in the study group is plotted as the standard equivalent concentration. Error bars represent the 95% CI of GMT.

[0102] Figure 27C Surgical Model of Chronic Pain: Seven dogs in two groups were treated with cNGF-CMV-Ntt830-E4 VLP vaccine and placebo (water for injection), respectively. Treatment was performed approximately 21 and 42 days prior to the implementation of the experimental model of chronic pain caused by right-sided stiff joint instability due to surgery. Lameness scores were assessed for each dog based on the typical course of the disease using a validated numerical rating scale. The numerical rating scale used to assess lameness was based on examiner perception and the scores were determined as follows: (0) no lameness; (1) barely detectable lameness for most of the observation period; (2) mild lameness with maximum weight-bearing capacity; (3) moderate lameness with minimum weight-bearing capacity; (4) severe lameness, with the animal using its paws (initiating walking movements and / or lightly touching the ground) but not bearing weight; (5) the animal with the most severe lameness, refusing to move and / or avoiding any contact between the inflamed paws and the ground. Lameness scores were measured for all dogs on day 217 of the study, approximately 175 days after surgery.

[0103] Figure 28A Pain assessment comparisons between groups. Group A: Vaccine candidate; Group B: Betinvelumab; Group C: Untreated at SD54-SD59 (at surgery), SD84, SD133, SD182, SD203, and SD217. Kruskal-Wallis statistical analysis and Dunn post-hoc test were used.

[0104] Figure 28B Limping assessment comparisons between groups. Group A: Vaccine candidate; Group B: Betinvelumab; Group C: Untreated at SD54-SD59 (at surgery), SD84, SD133, SD182, SD203, and SD217. Kruskal-Wallis statistical analysis and Dunn post-hoc test were used. Invention Details Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Specific, preferred, and / or highly preferred examples described and disclosed herein should be applied to all aspects and other specific, preferred, and / or highly preferred examples, whether or not specifically mentioned again or whether repetition is avoided for the sake of brevity. As used herein, the articles “a” and “an” refer to one or more (i.e., at least one) of the grammatical objects of the article. Unless the context clearly indicates otherwise, the term “or” as used herein should be understood to mean “and / or”.

[0106] Virus-like particles (VLPs): As used herein, the term "virus-like particle (VLP)" refers to a non-replicating or non-infectious particle, preferably a non-replicating and non-infectious viral particle, or a non-replicating or non-infectious structure, preferably a non-replicating and non-infectious structure similar to a viral particle, preferably a viral capsid. As used herein, the term "non-replicating" means that the genome contained in the VLP cannot be replicated. As used herein, the term "non-infectious" means that it cannot be introduced into a host cell. Virus-like particles according to the invention are non-replicating and non-infectious due to the lack of all or part of the viral genome or genome function. Virus-like particles according to the invention may contain nucleic acids different from their genome. Recombinant virus-like particles typically contain host cell-derived RNA. Typical and preferred specific examples of virus-like particles according to the invention are viral capsids composed of polypeptides of the invention. Virus-like particles are typically macromolecular assemblies composed of viral coat proteins, typically comprising 60, 120, 180, 240, 300, 360, or more than 360 protein subunits per virus-like particle. Typically and preferably, the interaction of these subunits leads to the formation of a viral capsid or virus-like protein shell structure with an inherently repetitive organization. A characteristic of virus-like particles is their high degree of order and the repetitive arrangement of their subunits.

[0107] Cucumber mosaic virus (CMV) virus-like particles: The term "CMV virus-like particle" or "CMV VLP" refers to a virus-like particle that contains at least one CMV polypeptide or is preferably substantially composed of it. Preferably, the CMV virus-like particle contains the CMV polypeptide as the major protein component of the capsid structure, and even more preferably as the sole protein component of the capsid structure. Typically and preferably, the CMV virus-like particle resembles the capsid structure of CMV. CMV virus-like particles are non-replicating and / or non-infectious, and at least lack one or more genes encoding the CMV replication mechanism, and typically also lack one or more genes encoding one or more proteins responsible for viral attachment to or entry into the host. This definition also includes virus-like particles in which the aforementioned one or more genes are still present but inactive. A preferred method for rendering CMV virus-like particles non-replicating and / or non-infectious is by physical or chemical inactivation (e.g., UV irradiation, formaldehyde treatment). Preferably, the CMV VLP lacks one or more genes encoding the CMV replication mechanism, and also lacks one or more genes encoding one or more proteins responsible for viral attachment to or entry into the host. More preferably, the non-replicating and / or non-infectious virus-like particles are obtained through recombinant gene technology. The virus-like particles generated from the recombinant CMV according to the invention typically and preferably do not contain a viral genome. Preferably, the CMV VLP is a macromolecular assembly composed of CMV capsid proteins or CMV polypeptides, typically comprising 180 capsid protein or CMV polypeptide subunits per VLP. Typically and preferably, as used herein, the CMV VLP comprises, is substantially composed of, or is alternatively composed of, at least one CMV polypeptide comprising or preferably comprises: (i) an amino acid sequence of a CMV capsid protein; or (ii) a mutant amino acid sequence, wherein the mutant amino acid sequence has at least 90%, preferably at least 95%, more preferably at least 98%, and even more preferably at least 99% sequence identity with the CMV capsid protein.

[0108] Modified CMV virus-like particles (VLPs): The term "modified CMV virus-like particle" refers to a virus-like particle comprising at least one modified CMV polypeptide, at least one chimeric CMV polypeptide, or at least one antigenic CMV fusion polypeptide as defined and described herein. Typically and preferably, the modified CMV VLP resembles the capsid structure of CMV. The modified CMV VLP is non-replicating and / or non-infectious, and at least lacks one or more genes encoding CMV replication mechanisms, and typically also lacks one or more genes encoding one or more proteins responsible for viral attachment to or entry into the host. This definition also includes modified virus-like particles in which the aforementioned one or more genes are still present but inactive. Preferably, the non-replicating and / or non-infectious modified virus-like particles are obtained through recombinant gene technology and typically and preferably do not contain a viral genome. Preferably, the modified CMV VLP is a macromolecular assembly composed of modified CMV peptides as described herein, and typically and preferably comprises 180 such protein subunits, each VLP comprising a modified CMV peptide, a chimeric peptide, or an antigenic CMV fusion peptide. Thus, in a preferred embodiment, the modified CMV VLP comprises, preferably, 180 modified CMV peptides. In another preferred embodiment, the modified CMV VLP comprises, preferably, 180 chimeric CMV peptides. In yet another preferred embodiment, the modified CMV VLP is a mosaic fusion CMV VLP comprising at least one, preferably multiple, antigenic CMV fusion peptides.

[0109] Mosaic fusion CMV VLP: The term "mosaic fusion CMV VLP" refers to a modified CMV VLP as defined herein and comprising at least one, preferably multiple, antigenic CMV fusion peptides as defined herein. Virus-like particles comprising peptides of more than one species and type are called mosaic VLPs. Thus, in another preferred embodiment, the composition comprises, preferably consists of, such a mosaic fusion CMV VLP, comprising at least one, preferably multiple, antigenic CMV fusion peptides as defined herein, and further comprising peptides selected from all CMV peptides as defined herein, modified CMV peptides, and CMV proteins.

[0110] Polypeptide: As used herein, the term "polypeptide" refers to a polymer composed of amino acid monomers linearly linked by amide bonds (also known as peptide bonds). It refers to an amino acid molecular chain, not a product of a specific length. Therefore, the definition of a polypeptide includes peptides, dipeptides, tripeptides, oligopeptides, and proteins. As used herein, typically and preferably, the term "polypeptide" should also refer to a polypeptide as previously defined and encompass modifications such as post-translational modifications, including but not limited to glycosylation. In a preferred specific example, as used herein, the term "polypeptide" should refer to a polypeptide as previously defined and exclude modifications such as post-translational modifications, such as glycosylation. Specifically, the biologically active peptide may subsequently undergo such modifications, such as glycosylation, even in vivo, for example, through bacteria.

[0111] Cucumber mosaic virus (CMV) polypeptide, CMV polypeptide: As used herein, the term "cucumber mosaic virus (CMV) polypeptide" refers to a polypeptide comprising or preferably consisting of: (i) the amino acid sequence of the coat protein of cucumber mosaic virus (CMV); or (ii) a mutant amino acid sequence wherein the mutant amino acid sequence and the CMV coat protein exhibit at least 90%, preferably at least 91%, 92%, 93% or 94%, more preferably at least 95%, even more preferably at least 98%, and still more preferably at least 99% sequence identity. Typically and preferably, the CMV polypeptide is capable of self-assembling into CMV virus-like particles after expression.

[0112] Cucumber mosaic virus (CMV) coat protein (CP): As used herein, the term "cucumber mosaic virus (CMV) coat protein (CP)" refers to the coat protein of naturally occurring cucumber mosaic viruses. Due to the extremely wide host range of cucumber mosaic viruses, many different strains and isolates of CMV are known. The coat protein sequences of these strains and isolates have been determined and are known to those skilled in the art. The sequences of the CMV coat protein (CP) are described in and can be retrieved from known databases such as Genbank, www.dpvweb.net, or www.ncbi.nlm.nih.gov / protein / . Specific examples of the CMV CP are described on page 12, line 8 to page 13, line 25 of WO 2016 / 062720, the disclosure of which is expressly incorporated herein by reference. Very preferred and specific examples of the CMV coat protein are provided in SEQ ID NO:39. Therefore, preferably, as used herein, the term "coat protein of cucumber mosaic virus (CMV)" refers to the amino acid sequence of the CMV coat protein, wherein the amino acid sequence comprises or preferably consists of the following: SEQ ID NO:39 or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 91%, 92%, 93% or 94%, even more preferably at least 95%, still more preferably at least 96% or 97%, still more preferably at least 98%, and still even more preferably at least 99% sequence identity with SEQ ID NO:39.

[0113] It is noteworthy that this viral strain and its isolates exhibit highly similar capsid protein sequences across different protein domains (including the N-terminus of the capsid protein). Specifically, 98.1% of all fully sequenced CMV isolates share over 85% sequence identity in the first 28 amino acids of their capsid protein sequences, and 79.5% of all fully sequenced CMV isolates still share over 90% sequence identity in the first 28 amino acids of their capsid protein sequences.

[0114] Modified CMV peptides: As used herein, the term "modified CMV peptide" refers to a CMV peptide comprising, or preferably composed of, a CMV peptide and a helper T cell epitope. Typically, modified CMV peptides are capable of self-assembling into CMV virus-like particles after expression. Preferably, the modified CMV peptide is a recombinant modified CMV peptide capable of self-assembling into CMV virus-like particles after expression in *E. coli*.

[0115] Chimeric CMV peptide: As used herein, the term "chimeric CMV peptide" refers to a peptide as defined herein and according to the invention, comprising, preferably composed of, a CMV peptide, wherein the CMV peptide is modified as defined and described herein to comprise a peptide comprising a segment of negatively charged, independently selected, continuous amino acids of aspartic acid or glutamic acid, preferably composed of a segment of such continuous negatively charged amino acids, and the CMV peptide is optionally further modified to include helper T cell epitopes, all components as defined and described herein. Typically and preferably, the chimeric CMV peptide is capable of self-assembling into modified CMV virus-like particles after expression. Thus, in a preferred embodiment, the chimeric CMV peptide is capable of self-assembling into modified CMV virus-like particles, typically and preferably after expression. Preferably, the chimeric CMV peptide is a recombinant modified CMV peptide capable of self-assembling into CMV virus-like particles after expression in *E. coli*. Typically and preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, wherein the substituted N-terminal region of the CMV polypeptide consists of 5 to 15 consecutive amino acids. Preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, and wherein the substituted N-terminal region of the CMV polypeptide consists of 5 to 15 consecutive amino acids, more preferably 9 to 14, 9 to 13, or 10 to 13 consecutive amino acids, more preferably 11 to 13 consecutive amino acids, and most preferably 11, 12, or 13 consecutive amino acids.

[0116] Antigenic CMV Fusion Peptide: As used herein, the term "antigenic CMV fusion peptide" refers to a peptide comprising a CMV peptide, preferably composed of CMV peptides, wherein the CMV peptide is modified to comprise an antigenic peptide as defined and described herein, and optionally further modified to comprise a helper T cell epitope. Typically and preferably, the antigenic CMV peptide is capable of self-assembling into modified CMV virus-like particles after expression. Thus, in a preferred embodiment, the antigenic CMV fusion peptide is capable of self-assembling, typically and preferably, after expression (typically and preferably after expression in *E. coli*) to form modified CMV virus-like particles. In another preferred embodiment, the antigenic CMV fusion peptide is co-expressed with a peptide selected from modified CMV peptides, CMV peptides, and CMV proteins as defined herein, to form a mosaic fusion CMV (CMV) through self-assembly, typically and preferably, after expression (typically and preferably after expression in *E. coli*). Typically and preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, wherein the substituted N-terminal region of the CMV polypeptide consists of 5 to 15 consecutive amino acids. Preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, and wherein the substituted N-terminal region of the CMV polypeptide consists of 5 to 15 consecutive amino acids, more preferably 9 to 14, 9 to 13, or 10 to 13 consecutive amino acids, more preferably 11 to 13 consecutive amino acids, and most preferably 11, 12, or 13 consecutive amino acids.

[0117] N-terminal region of CMV peptide: As used herein, the term "N-terminal region of CMV peptide" refers to the N-terminus of the CMV peptide, and particularly to the N-terminus of the CMV capsid protein, or the N-terminal region of the CMV peptide or the CMV capsid protein. However, if the CMV peptide or the capsid protein contains an N-terminal methionine residue, the N-terminal region begins at the second amino acid at the N-terminus of the CMV peptide or the CMV capsid protein. Preferably, in the case where the CMV peptide or the capsid protein contains an N-terminal methionine residue, from a practical point of view, the start codon encoding the methionine is typically deleted and added to the N-terminus of the helper T (Th) cell epitope. More preferably, one, two, or three other amino acids may be optionally inserted between the methionine and the helper T cell epitope, preferably one amino acid, for cloning purposes.

[0118] Recombinant polypeptide: In the context of this invention, the term "recombinant" as used in the context of a polypeptide refers to a polypeptide obtained by a method comprising at least one step of recombinant DNA technology. Typically and preferably, the recombinant polypeptide is produced in a prokaryotic expression system. It will be apparent to those skilled in the art that the recombinant polypeptide produced by expression in a prokaryotic expression system (such as *E. coli*) may contain an N-terminal methionine residue. The N-terminal methionine residue is typically cleaved from the recombinant polypeptide in the expression host during the maturation of the recombinant polypeptide. However, the cleavage of the N-terminal methionine may be incomplete. Therefore, the preparation of the recombinant polypeptide may comprise a mixture of polypeptides having an N-terminal methionine residue and polypeptides otherwise identical without an N-terminal methionine residue. Typically and preferably, the recombinant polypeptide formulation comprises less than 10%, more preferably less than 5%, and still more preferably less than 1%, of the recombinant polypeptide having an N-terminal methionine residue.

[0119] Recombinant modified virus-like particles: In the context of this invention, the term "recombinant modified virus-like particles" refers to modified virus-like particles (VLPs) obtained by a method comprising at least one step of recombinant DNA technology.

[0120] Mutant amino acid sequence: The term "mutant amino acid sequence" refers to an amino acid sequence obtained by introducing a set of defined mutations into an amino acid sequence to be mutated. In the context of this invention, the amino acid sequence to be mutated is typically and preferably the amino acid sequence of a CMV outer shell protein. Therefore, the mutant amino acid sequence differs from the amino acid sequence of the CMV outer shell protein by at least one amino acid residue, wherein the mutant amino acid sequence shows at least 90% sequence identity with the amino acid sequence to be mutated. Typically and preferably, the mutant amino acid sequence shows at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence to be mutated. Preferably, the mutant amino acid sequence differs from the sequence to be mutated by at most 11, 10, 9, 8, 7, 6, 4, 3, 2, or 1 amino acid residue, more preferably, the difference is selected from insertions, deletions, and amino acid exchanges. Preferably, the mutant amino acid sequence differs from the amino acid sequence of the CMV outer shell protein by at least one amino acid, wherein preferably, the difference is an amino acid exchange.

[0121] The terms "corresponding" or "corresponds" used herein to describe the relationship between specific positions of amino acid residues within peptide and amino acid sequences, respectively, refer to the correspondence between an amino acid residue position in one amino acid sequence and a specified amino acid residue in another amino acid sequence. This can be identified through sequence alignment, typically and preferably using the BLASTP algorithm, and most preferably using standard settings. Typical and preferred standard settings are: expected threshold: 10; word length: 3; maximum match within the query range: 0; matrix: BLOSUM62; empty space cost: 11 for existence, 1 for extension; compositional adjustment: conditional compositional score matrix adjustment.

[0122] Sequence Identification: The sequence identity of the two specified amino acid sequences is determined based on the alignment of the two sequences. Algorithms for determining sequence identity are available to those skilled in the art. Preferably, the sequence identity of the two amino acid sequences is determined using a publicly available computer homology program, such as the "BLAST" program (http: / / blast.ncbi.nlm.nih.gov / Blast.cgi) or "CLUSTALW" (http: / / www.genome.jp / tools / clustalw / ), and hereby preferably using the "BLAST" program provided on the NCBI homepage http: / / blast.ncbi.nlm.nih.gov / Blast.cgi, using the preset settings provided therein. Typical and preferred criterion settings are: expected threshold: 10; word length: 3; maximum match within the query range: 0; matrix: BLOSUM62; empty space cost: 11 for existence, 1 for extension; compositional adjustment: conditional compositional scoring matrix adjustment.

[0123] Amino acid exchange: The term "amino acid exchange" refers to the exchange of a specified amino acid residue in an amino acid sequence with any other amino acid residue having a different chemical structure, preferably with another protein-type amino acid residue. Therefore, amino acid exchange does not change the total number of amino acids in the amino acid sequence compared to the insertion or deletion of an amino acid.

[0124] As used herein, the term "isoelectric point," abbreviated as pI, refers to the pH at which a molecule carries no net charge or is statistically electrically neutral. Specifically, the term "isoelectric point" is used herein to refer to the pH at which an antigen used in this invention and composed of amino acids carries no net charge or is statistically electrically neutral. Such antigens carry a net positive charge at pH below their pI and a net negative charge at pH above their pI. Typically and preferably, when referring to pI values, and particularly when referring to the pI values ​​of the antigens of the present invention and the antigens of this disclosure, the pI values ​​are determined by inputting the primary amino acid sequences of the specific protein and antigen into the ExPASy Compute pI / MW tool described by Gasteiger et al. (Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, MR, Appel, RD, & Bairoch, A., Protein Identification and Analysis Tools on the ExPASy Server, available in John M. Walke (ed.): The Proteomics Protocols Handbook, Humana Press (2005)). Therefore, the ExPASy Compute pI / MW tool mentioned herein refers to the tool described by Gasteiger et al. This tool calculates the theoretical isoelectric point pI and Mw of the specified Swiss-Prot / TrEMBL entry or the user-input amino acid sequence. Protein pI is calculated using the amino acid pK values ​​described by Bjellqvist et al., which is defined by examining the migration of peptides in a fixed pH gradient gel environment containing 9.2M and 9.8M urea, between pH 4.5 and 7.3, at 15°C or 25°C (Bjellqvist, B. et al., 1993, Electrophoresis 14:1023-1031; Bjellqvist, B. et al., 1994, Electrophoresis 15:529-539).

[0125] Epitope: The term "eptope" refers to a continuous or discontinuous portion of a polypeptide or antigen that can be specifically bound by an antibody or by a T-cell receptor within the background of MHC molecules. In the case of antibodies, specific binding excludes non-specific binding, but does not necessarily exclude cross-reactivity. Epitopes typically contain 5 to 20 amino acids in a spatial conformation unique to the antigen site.

[0126] Helper T (Th) cell epitopes: The term “helper T cell epitope or Th cell epitope” as used herein and interchangeably refers to an epitope that can be recognized by helper Th cells. Typically and preferably, as used herein, the term “Th cell epitope” refers to a Th cell epitope that can bind to at least one, preferably more than one, class II MHC molecule. The simplest way to determine whether a peptide sequence is a Th cell epitope is to measure the peptide’s ability to bind to each class II MHC molecule. This can be measured based on the peptide’s ability to compete with known Th cell epitope peptides for binding to class II MHC molecules. Representative selections of HLA-DR molecules are described, for example, in Alexander J et al., Immunity (1994) 1:751-761. The affinity of a Th cell epitope for a class II MHC molecule should be at least 10. -5M. A representative group of class II MHC molecules present in different individuals is provided in Panina-Bordignon P et al., Eur J Immunol (1989) 19:2237-2242. Therefore, as used herein, the term "Th cell epitope" preferably refers to a Th cell epitope that generates a measurable T cell response after immunization and booster immunization. Furthermore, and more preferably, as used herein, the term "Th cell epitope" preferably refers to a Th cell epitope capable of binding with an affinity of at least 500 nM to at least one, preferably at least two, and even more preferably at least three DR alleles selected from DR1, DR2w2b, DR3, DR4w4, DR4w14, DR5, DR7, DR52a, DRw53, and DR2w2a; and preferably selected from DR1, DR2w2b, DR4w4, DR4w14, DR5, DR7, DRw53, and DR2w2a (as described in Alexander J et al., Immunity (1994) 1:751-761 and the references cited therein); the preferred binding analysis for assessing the affinity is the binding analysis described in Sette A et al., J Immunol (1989) 142:35-40. In an even more preferred manner, as used herein, the term "Th cell epitope" refers to a Th cell epitope capable of binding with an affinity of at least 500 nM to at least one, preferably at least two, and even more preferably at least three DR alleles selected from DR1, DR2w2b, DR4w4, DR4w14, DR5, DR7, DRw53, DR2w2a (as described in Alexander J et al., Immunity (1994) 1:751-761 and the references cited therein); the preferred binding analysis for assessing the affinity is the binding analysis described in Sette A et al., J Immunol (1989) 142:35-40. Th cell epitopes have been described and are known to those skilled in the art, such as Alexander J et al., Immunity (1994) 1:751-761; Panina-Bordignon P et al., Eur J Immunol (1989) 19:2237-2242; Calvo-Calle JM et al., J Immunol (1997) 159:1362-1373; and Valmori D et al., J Immunol (1992) 149:717-721.

[0127] Amino acid linker: As used herein, the term "amino acid linker" refers to a linker composed entirely of amino acid residues. The amino acid residues of an amino acid linker are naturally occurring amino acids or non-natural amino acids, or mixtures thereof, known in the art as being entirely L- or entirely D-configured. Preferably, the amino acid residues of an amino acid linker are naturally occurring amino acids or mixtures thereof being entirely L- or entirely D-configured. In a preferred embodiment, the amino acid linker is composed of naturally occurring α-amino acids (all in their L-configuration).

[0128] G-Linker: As used herein, the term "G-linker" refers to an amino acid linker consisting solely of glycine amino acid residues. The G-linkers according to the invention comprise at least two glycine residues and at most ten glycine residues.

[0129] GS-linker: As used herein, the term "GS-linker" refers to an amino acid linker consisting only of glycine and serine amino acid residues. The GS-linker according to the invention comprises at least one glycine and at least one serine residue. Typically and preferably, the GS-linker is up to 30 amino acids in length.

[0130] GS*-linker: As used herein, the term "GS*-linker" refers to an amino acid linker comprising at least one glycine, at least one serine, and at least one amino acid residue selected from Thr, Ala, Lys, and Cys. Typically and preferably, the GS*-linker is up to 30 amino acids in length.

[0131] As used herein, the term "amino acid" refers to an organic compound containing an amino group (-NH2) and a carboxylic acid group (-COOH) and their zwitterions, and typically and preferably, contains the side chain characteristic of each amino acid. The term "amino acid" typically and preferably includes naturally occurring amino acids, such as protein amino acids (produced via RNA translation), non-protein amino acids (produced through other metabolic mechanisms, such as post-translational modifications), standard or typical amino acids (directly encoded by codons in the genetic code), and non-standard or atypical amino acids (not directly encoded by the genetic code). Naturally occurring amino acids include both eukaryotic and eukaryotic amino acids. As used herein, the term "amino acid" also includes chemically synthesized non-natural amino acids; amino acids such as alpha-(α-), beta-(β-), gamma-(γ-), and delta-(δ-), and mixtures thereof in any ratio; and, where applicable, mixtures thereof such as alpha-(α-) amino acids, any isomers of amino acids, i.e., their D-stereoisomers and L-stereoisomers (alternatively described according to (R) and (S) nomenclature), and in any ratio (including a 1:1 racemic ratio). The terms "D-stereoisomer," "L-stereoisomer," "D-amino acid," or "L-amino acid" refer to the chiral α-carbon of an amino acid. In a preferred embodiment, the term amino acid refers to an α-amino acid, preferably a naturally occurring α-amino acid, and more preferably a naturally occurring α-amino acid in the L-configuration.

[0132] Association: As used herein, the term “associated” or “association” refers to all possible ways in which two molecules are linked together, preferably through chemical interactions. Chemical interactions include both covalent and non-covalent interactions. Typical examples of non-covalent interactions are ionic interactions, hydrophobic interactions, or hydrogen bonds, while covalent interactions are based on covalent bonds such as esters, ethers, phosphate esters, carbon-phosphorus bonds, carbon-sulfur bonds such as thioethers, or imide bonds.

[0133] First attachment site: As used herein, the term "first attachment site" refers to an element naturally present in or artificially added to a virus-like particle (VLP) that can connect to a second attachment site. The first attachment site is preferably a protein, polypeptide, amino acid, peptide, sugar, polynucleotide, natural or synthetic polymer, secondary metabolite, or compound such as biotin, fluorescein, retinol, digoxigenin, metal ions, phenylmethylsulfonyl fluoride, or a chemically reactive group such as amino, carboxyl, mercapto, hydroxyl, guanidinyl, histidine, or combinations thereof. A preferred specific example of a chemically reactive group as a first attachment site is an amino group of an amino acid residue, preferably an amino group of a lysine residue's side chain. In another preferred specific example, the first attachment site is a naturally occurring amino acid residue in the VLP. In yet another preferred specific example, the first attachment site is a non-naturally occurring amino acid residue in the VLP. The first attachment site is typically located on a surface, and preferably on the outer surface of the VLP. Multiple first attachment sites are present on the surface, preferably on the outer surface of the VLP, and typically in a repeating configuration. In a preferred embodiment, the first attachment site is linked to the VLP via at least one covalent bond, preferably via at least one peptide bond. In another preferred embodiment, the first attachment site is naturally present in the VLP. Alternatively, in a preferred embodiment, the first attachment site is artificially added to the VLP. In a very preferred embodiment, the first attachment site is the amino group of a lysine residue in the amino acid sequence of the VLP polypeptide. In another very preferred embodiment, the first attachment site is the amino group of a lysine residue in the amino acid sequence of the VLP polypeptide, wherein the lysine residue is naturally present in the VLP polypeptide. In another very preferred embodiment, the first attachment site is the amino group of a lysine residue in the amino acid sequence of the VLP polypeptide, wherein the lysine residue is not naturally present in the VLP polypeptide but is artificially added, for example, by adding it to the VLP polypeptide or by substituting a naturally present amino acid in the VLP polypeptide. In another very preferred embodiment, the first attachment site is a naturally present amino acid residue in the VLP polypeptide. In another highly preferred embodiment, the first attachment site is an amino acid residue in the VLP polypeptide that is not naturally present but artificially added, for example by adding it to the VLP polypeptide or by replacing the naturally present amino acid in the VLP polypeptide.

[0134] Second attachment site: As used herein, the term "second attachment site" refers to an element naturally present in the antigen or artificially added to the antigen and capable of connecting to the first attachment site. The second attachment site of an antigen is preferably a protein, polypeptide, peptide, amino acid, sugar, polynucleotide, natural or synthetic polymer, secondary metabolite, or compound such as biotin, fluorescein, retinol, digoxin, metal ions, phenylmethylsulfonyl fluoride, or a chemically reactive group such as amino, carboxyl, thiol, hydroxyl, guanidinyl, histidine, or combinations thereof. A preferred specific example of a chemically reactive group as a second attachment site is a thiol group, preferably a thiol group in a cysteine ​​residue. In another preferred specific example, the second attachment site is a naturally present amino acid residue in the antigen. In yet another preferred specific example, the second attachment site is a non-naturally present amino acid residue in the antigen. Therefore, the term "antigen having at least one second attachment site" refers to a construct comprising an antigen and at least one second attachment site. However, particularly for second attachment sites not naturally present in the antigen, such constructs typically and preferably further include a "connector." In another preferred embodiment, the second attachment site is linked to the antigen via at least one covalent bond, preferably via at least one peptide bond. In another preferred embodiment, the second attachment site is naturally present in the antigen. In another more preferred embodiment, the second attachment site is artificially added to the antigen via a linker, wherein the linker comprises or is alternatively composed of cysteine. Preferably, the linker is fused to the antigen via a peptide bond. In a very preferred embodiment, the second attachment site is a thiol group, preferably a thiol group of a cysteine ​​residue. Preferably, the linker is fused to the antigen via a peptide bond. In another very preferred embodiment, the second attachment site is an amino acid residue naturally present in the antigen. In another very preferred embodiment, the second attachment site is an amino acid residue not naturally present in the antigen but artificially added, for example by adding it to the antigen or by substituting an amino acid naturally present in the antigen.

[0135] Linked: As used herein, the term "linked" or "linkage" refers to all possible ways in which at least one first attachment site is connected to at least one second attachment site, preferably through chemical interaction. Chemical interaction includes both covalent and non-covalent interactions. Typical examples of non-covalent interactions are ionic interactions, hydrophobic interactions, or hydrogen bonds, while covalent interactions are based on covalent bonds such as esters, ethers, phosphate esters, carbon-phosphorus bonds, carbon-sulfur bonds such as thioethers, or imide bonds. In some preferred embodiments, the first attachment site and the second attachment site are linked by at least one covalent bond, preferably by at least one non-peptide bond, and even more preferably exclusively by a non-peptide covalent bond. In other preferred embodiments, the first attachment site and the second attachment site are linked by at least one covalent bond, preferably by at least one peptide bond, and even more preferably exclusively by a peptide covalent bond. However, as used herein, the term "linkage" should not only refer to a direct link between at least one first attachment site and at least one second attachment site, but also, alternatively and preferably, to an indirect link between at least one first attachment site and at least one second attachment site via an intermediate molecule, typically and preferably via at least one heterobifunctional crosslinker. In other preferred embodiments, the first and second attachment sites are linked by at least one covalent bond, preferably by at least one peptide bond, and even more preferably by peptide bonds only. In one preferred embodiment, the first and second attachment sites are linked by a peptide bond representing a direct peptide bond between the first and second attachment sites. In another preferred embodiment, the first and second attachment sites are linked by peptide bonds representing the peptide bonds between the first and second attachment sites, via amino acid linkers as described herein. If an antigen (preferably an antigenic polypeptide) is fused into a VLP polypeptide, two direct peptide bonds may exist between the first and second attachment sites, or one or two peptide bonds may exist between the first and second attachment sites, linked by one or two identical or different amino acid linkers as described herein.

[0136] Linker: As used herein, a "linker" connects a second attachment site to an antigen, or contains or consists of a second attachment site. Preferably, as used herein, a "linker" contains a second attachment site, typically and preferably as an amino acid residue, more preferably as a cysteine ​​residue. A preferred linker is one containing at least one amino acid residue, or even more preferably, a linker consisting entirely of amino acid residues. The amino acid residues of the linker are preferably composed of naturally occurring all-L or all-D amino acids or non-natural amino acids or mixtures thereof known in the art. More preferred specific examples of linkers according to the invention are molecules containing thiol or cysteine ​​residues, and therefore, such molecules are also covered within the scope of the invention. Other linkers suitable for the invention are molecules containing C1-6 alkyl-, cycloalkyl (such as cyclopentyl or cyclohexyl), cycloalkenyl, aryl, or heteroaryl moieties. Furthermore, linkers preferably containing C1-C6 alkyl-, cycloalkyl- (C5, C6), aryl, or heteroaryl moieties and other amino acids may also be used as linkers of the invention and should be covered within the scope of the invention. The linker and the antigen are preferably linked by at least one covalent bond, more preferably by at least one peptide bond.

[0137] Antigen: The antigen of the present invention is a nerve growth factor (NGF) antigen. Therefore, as used herein, the term "antigen" refers to a nerve growth factor (NGF) antigen that is typically and preferably capable of being bound by an antibody or a T-cell receptor (TCR) upon presentation of an MHC molecule.

[0138] Antigenic polypeptide: As used herein, the term "antigenic polypeptide" refers to a nerve growth factor (NGF) antigen as defined herein, and is a polypeptide contained in an antigenic CMV fusion polypeptide as defined herein. The terms "antigenic polypeptide" and "antigenic NGF polypeptide" may be used interchangeably herein.

[0139] Ordered and repetitive antigen arrays: As used herein, the term "ordered and repetitive antigen array" refers to a repeating pattern of antigens, typically and preferably characterized by a highly homogeneous spatial arrangement of antigens relative to the modified CMV VLP. In a specific embodiment of the invention, the repeating pattern may be a geometric pattern. Certain specific embodiments of the invention (such as antigens linked to modified CMV VLPs) are typical and preferred examples of suitable ordered and repetitive antigen arrays, further having strictly repeating antigen sub-sequences, preferably with a spacing of 1 to 30 nanometers, more preferably 2 to 15 nanometers, even more preferably 2 to 10 nanometers, even more preferably 2 to 8 nanometers, and even more preferably 1.6 to 7 nanometers.

[0140] Coupling efficiency: The coupling efficiency of virus-like particles to a specific antigen is determined by SDS-PAGE of the coupling reaction. The intensity of the Coumarin blue staining bands corresponding to the components of the coupling reaction is determined by densitometric analysis and used to calculate the coupling efficiency. The coupling efficiency is defined as (i) the ratio of the amount of VLP peptide coupled to the antigen to (ii) the total amount of VLP peptides. Typically and preferably, the coupling efficiency is at least 5%, 10%, preferably at least 15%, more preferably at least 20%, 25%, or at least 30%, and even more preferably at least 35% or at least 40%. The coupling efficiency can also be expressed as the total number of antigens linked to the modified CMV VLP. The coupling efficiency may depend on the nature of the antigen, and the total number of antigens linked to the modified CMV VLP is typically and preferably at least 5, at least 7, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, and at least 50 antigens.

[0141] Nerve growth factor (NGF) antigen: As used herein, the term "nerve growth factor (NGF) antigen" refers to a biologically active NGF antigen. As used herein, a biologically active NGF antigen generally refers to the ability to bind to the NGF high-affinity receptor TrkA and / or activate the NGF receptor signaling pathway. When administered to canines in the form of the compositions of the present invention, the NGF antigen typically and preferably induces the canines to produce anti-NGF antibodies, wherein the anti-NGF antibodies are preferably capable of neutralizing the biological activity of nerve growth factor (NGF) in an in vitro assay, preferably as described herein (see Example 6). As used herein and when referring to NGF antigen, the term "biological activity" refers to the activity of the NGF antigen in a cell proliferation assay, preferably based on an NGF-dependent human erythroleukemia TF-1 cell line, and more preferably, under conditions substantially as described herein in Example 6. Furthermore, NGF antigen typically refers to a polypeptide exhibiting biological activity comprising, preferably, the following: an amino acid sequence of canine or feline nerve growth factor or a corresponding heterologous homolog from any other species, preferably from a non-human animal. Specifically, it refers to a polypeptide comprising, preferably, an amino acid sequence of canine or feline nerve growth factor or a corresponding heterologous homolog from any other species, preferably from a non-human animal, or a polypeptide having at least 90%, preferably at least 92%, more preferably at least 95%, and even more preferably at least 98% sequence identity with the amino acid sequence of canine or feline nerve growth factor or a corresponding heterologous homolog from any other species, preferably from a non-human animal. Preferred NGF antigens from various animal species are canine NGF (cNGF), feline NGF (fNGF), equine NGF (eNGF), bovine NGF (bNGF), and suidae NGF (pNGF), preferably canine NGF (cNGF) or feline NGF (fNGF), and the NGF antigen comprises, preferably consists of, the following: polypeptides of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58, or amino acid sequences having at least 90%, preferably at least 92%, more preferably at least 95%, and even more preferably at least 98% sequence identity with any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58.

[0142] Endogenous or autologous NGF protein: As used herein, the term "endogenous or autologous NGF protein" refers to the NGF protein in an animal receiving treatment according to the present invention. In this context, the terms "autologous NGF" and "endogenous NGF" are used interchangeably.

[0143] CMV VLP composition containing NGF: As used herein, the term “CMV VLP composition containing NGF” refers to a composition comprising: (a) a CVM VLP containing at least one attachment site and (b) at least one NGF antigen containing at least one second attachment site, wherein (a) and (b) are connected via the at least one first attachment site and the at least one second attachment site.

[0144] Adjuvants: As used herein, the term "adjuvant" refers to an immune response stimulant and / or a substance that typically allows the formation of a reservoir in the host, which, when combined with the compositions of the present invention, vaccines, or pharmaceutical compositions, enables a stronger immune response. Different types of adjuvants with different mechanisms of action have been described and have been able to enhance antigen-specific antibody responses (Pulendran B et al., 2021, Nature Reviews Drug Discovery 20:454-475). Typical and preferred adjuvants are mineral salts (e.g., aluminum hydroxide, aluminum phosphate), microcrystalline tyrosine, emulsions, microparticles, saponins (Quil A), cytokines, immune enhancers, microbial components / products, liposomes, complexes, and mucosal adjuvants, which are known and described, for example, in Adjuvant Compendium NIAID and VAC (nih.gov) or Aguilar et al. (Aguilar JC et al., 2007, Vaccine 25:3752-3762), Gerdts (Gerdts V, 2015, Berliner und Münchener Tierärztliche Wochenschrift 128:456-463) and Pasquale et al. (Pasquale et al., 2015, Vaccines 3:320-343). As used herein, the term "adjuvant" may also include mixtures of adjuvants. Virus-like particles are sometimes described as adjuvants. However, as used in the context of this application, the term "adjuvant" refers to an adjuvant that is not of the modified virus-like particles of this invention. Rather, "adjuvant" refers to another distinct component in the compositions, vaccines, or pharmaceutical compositions of this invention.

[0145] Immunostimulants: As used herein, the term "immunostimulant" refers to a substance capable of inducing and / or enhancing an immune response. As used herein, immunostimulants include, but are not limited to, Toll-like receptor activators and substances that induce cytokine secretion. Toll-like receptor activators include, but are not limited to, immunostimulatory nucleic acids, peptidoglycans, lipopolysaccharides, lipophosphoric acid, imidazoquinoline compounds, flagellins, and immunostimulatory organic substances such as paclitaxel.

[0146] Immunostimulating Nucleic Acid (ISS-NA): As used herein, the term "immunostimulating nucleic acid" refers to a nucleic acid capable of inducing and / or enhancing an immune response. Immunostimulating nucleic acids comprise ribonucleic acid, particularly deoxyribonucleic acid, wherein both ribonucleic acid and deoxyribonucleic acid can be double-stranded or single-stranded. Preferred ISS-NAs are deoxyribonucleic acid, more preferably single-stranded. Preferably, the immunostimulating nucleic acid contains at least one CpG motif comprising an unmethylated C. Very preferred immunostimulating nucleic acids contain at least one CpG motif, wherein the at least one CpG motif comprises at least one, preferably one, CG dinucleotide or preferably consists thereof, wherein the C is unmethylated. Preferably, but not necessarily, the CG dinucleotide is part of a palindromic sequence. The term immunostimulating nucleic acid also refers to a nucleic acid containing a modified base (preferably 4-bromo-cytosine). In the context of this invention, ISS-NAs capable of stimulating dendritic cells to produce IFN-α are particularly preferred. Immunostimulatory nucleic acids suitable for the purposes of this invention are described, for example, in WO2007 / 068747A1.

[0147] Oligonucleotide: As used herein, the term "oligonucleotide" refers to a nucleic acid sequence comprising two or more nucleotides, preferably about 6 to about 200 nucleotides, more preferably about 20 to about 100 nucleotides, and most preferably 20 to 40 nucleotides. Oligonucleotides are polynucleotides or polydeoxynucleotides and are preferably selected from (a) unmodified RNA or DNA, and (b) modified RNA or DNA. Modifications may include a backbone or nucleotide analogs. Oligonucleotides are preferably selected from the group consisting of: (a) single-stranded and double-stranded DNA; (b) DNA as a mixture of single-stranded and double-stranded regions; (c) single-stranded and double-stranded RNA; (d) RNA as a mixture of single-stranded and double-stranded regions; and (e) hybrid molecules comprising DNA and RNA, wherein the DNA and RNA are single-stranded or more preferably double-stranded or a mixture of single-stranded and double-stranded regions. Preferred nucleotide modifications / analytes are selected from the group consisting of: (a) peptide nucleic acids, (b) inosine, (c) triphenylmethylated bases, (d) thiophosphates, (e) alkyl thiophosphates, (f) 5-nitroindole deoxyribonucleoside, (g) 5-methyldeoxycytosine, and (h) 5,6-dihydro-5,6-dihydroxydeoxythymidine. Phosphophosphate-modified nucleotides are protected against degradation in cells or organisms and are therefore preferred nucleotide modifications. Unmodified oligonucleotides consisting entirely of phosphodiester-bound nucleotides typically have greater activity than modified nucleotides and are therefore generally preferred in the context of this invention. Most preferably, oligonucleotides consist entirely of phosphodiester-bound oligonucleotides, more preferably, said oligonucleotides are single-stranded. More preferably, oligonucleotides capable of stimulating the production of IFN-α by cells, preferably dendritic cells. Very preferred oligonucleotides capable of stimulating the production of IFN-α by cells are selected from type A CpG and type C CpG. More preferably, it is an RNA molecule that does not have a cap.

[0148] CpG motif: As used herein, the term "CpG motif" refers to a pattern of nucleotides comprising an unmethylated central CpG, i.e., an unmethylated CpG dinucleotide, wherein the C atoms are unmethylated; the central CpG is surrounded by at least one or two bases (preferably one or two nucleotides) located on either side of the central CpG (on the 3' and 5' sides). Typically and preferably, as used herein, a CpG motif comprises, or alternatively consists of, an unmethylated CpG dinucleotide and two nucleotides located at its 5' and 3' ends. Without being bound by theory, the bases flanking the CpG impart a significant portion of the activity to the CpG oligonucleotide.

[0149] Oligonucleotides containing unmethylated CpG: As used herein, the term "oligonucleotide containing unmethylated CpG" or "CpG" refers to an oligonucleotide containing at least one CpG motif, preferably an oligodeoxynucleotide. Thus, CpG contains at least one unmethylated cytosine and guanine dinucleotide. Preferred CpGs stimulate / activate vertebrate bone marrow-derived cells, for example, by promoting mitosis or inducing or increasing cytokine expression. For example, CpGs can be used to activate B cells, NK cells, and antigen-presenting cells such as dendritic cells, monocytes, and macrophages. Preferably, CpG refers to an oligodeoxynucleotide containing unmethylated cytosine and a guanosine linked at its 3' end, more preferably a single-stranded oligodeoxynucleotide, wherein the unmethylated cytosine and the guanosine are linked by a phosphate ester bond, preferably a phosphodiester bond or a thiophosphate bond, and more preferably a phosphodiester bond. CpGs may include nucleotide analogs, such as those containing phosphate thioester bonds, and may be double-stranded or single-stranded. Generally, double-stranded molecules are more stable in vivo, while single-stranded molecules have enhanced immune activity. Preferably, as used herein, a CpG is an oligonucleotide of at least about ten nucleotides in length and containing at least one CpG motif, more preferably, the CpG is 10 to 60 nucleotides in length, more preferably 15 to 50 nucleotides, still more preferably 20 to 40 nucleotides, still more preferably about 30 nucleotides, and most preferably exactly 30 nucleotides. A CpG may consist of methylated nucleotides and / or unmethylated nucleotides, wherein the at least one CpG motif contains at least one CG dinucleotide, wherein C is unmethylated. A CpG may also contain methylated and unmethylated sequence segments, wherein the at least one CpG motif contains at least one CG dinucleotide, wherein C is unmethylated. Very preferably, CpG refers to a single-stranded oligodeoxynucleotide containing an unmethylated cytosine and a guanosine linked at its 3' end, wherein the unmethylated cytosine and the guanosine are linked by a phosphodiester bond. CpG may include nucleotide analogs, such as those containing a thiophosphate bond, and may be double-stranded or single-stranded. Generally, phosphodiester CpGs are type A CpGs as shown below, while thiophosphate-stabilized CpGs are type B CpGs. In the context of this invention, type A CpGs are preferred.

[0150] Type A CpG: As used herein, the term "Type A CpG" or "Type D CpG" refers to an oligodeoxynucleotide (ODN) containing at least one CpG motif. Type A CpG preferentially stimulates T cell activation and dendritic cell maturation and can stimulate IFN-α production. In Type A CpG, the nucleotides of at least one CpG motif are linked by at least one phosphodiester bond. Type A CpG contains at least one phosphodiester bonded CpG motif, which may be flanked at its 5' end and / or preferably at its 3' end, and is linked by a phosphate-thioester-bound nucleotide. Preferably, the CpG motif, and herein preferably, the CG dinucleotide containing at least one, preferably two nucleotides, and its immediately adjacent flanking region, is composed of phosphodiester nucleotides. Preferably, Type A CpG consists entirely of phosphodiester (PO) bonded nucleotides. Typically and preferably, the poly-G motif comprises, or alternatively consists of, at least one guanosine (G), preferably at least three, at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 Gs, most preferably at least 10 Gs. Preferably, the type A CpG of the present invention comprises, or alternatively consists of, a palindromic sequence.

[0151] Encapsulation: As used herein, the term "encapsulation" refers to the state of the polyanionic macromolecule or immunostimulant relative to the core particle and the VLP, respectively. As used herein, the term "encapsulation" includes covalent (e.g., via chemical coupling) or non-covalent (e.g., ionic interactions, hydrophobic interactions, hydrogen bonds, etc.) binding. The term also includes the enclosure or partial enclosure of the polyanionic macromolecule. Therefore, the polyanionic macromolecule or immunostimulant can be encapsulated by the VLP in the absence of actual binding, particularly in the absence of covalent binding. In a preferred embodiment, at least one polyanionic macromolecule or immunostimulant is encapsulated within the VLP, most preferably in a non-covalent manner. When the immunostimulant is a nucleic acid (preferably DNA), the term encapsulation means that the nucleic acid cannot be hydrolyzed by nucleases, preferably not by deoxyribonucleases (e.g., deoxyribonuclease I or Benzonase), wherein preferably, the accessibility is analyzed as described in Examples 11-17 of WO2003 / 024481A2.

[0152] Effective Amount: As used herein, the term "effective amount" means the amount necessary to achieve or sufficient to achieve the desired biological effect. The effective amount of a composition (or, alternatively, a pharmaceutical composition) will be the amount that achieves this selected result, and such amount can be routinely determined by those skilled in the art. The effective amount can vary depending on the specific composition applied and the individual's body type. Those skilled in the art can determine the effective amount of a particular composition of the invention based on experience without excessive experimentation. Preferably, the term "effective amount" means an amount that (i) treats or prevents the specific disease or condition described herein; (ii) alleviates, improves, or eliminates one or more symptoms of the specific disease or condition described herein; or (iii) prevents or delays the onset of one or more symptoms of the specific disease or condition described herein.

[0153] Canids: As used in this article, the term "canids" refers to domestic dogs (dogs).

[0154] Veterinary compositions: As used herein, the term “veterinary composition” refers to a composition suitable for use on non-human animals.

[0155] NGF-related conditions: As used herein, the term "NGF-related condition" refers to any condition caused by any dysfunction or dysregulation of the NGF / TrkrA receptor pathway. NGF-related conditions include, but are not limited to, pain such as nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders (such as rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, seronegative (non-rheumatoid) arthropathy, non-articular rheumatoid diseases, and periarticular disorders), and pain associated with degenerative joint diseases (such as arthritis or osteoarthritis). Such pain can be acute, chronic, and / or refractory in nature.

[0156] Pain: As used herein, the term “pain” refers to pain of any etiology, including acute and chronic pain, and any pain with an inflammatory component. Examples of pain include nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders (such as rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, serologically negative (non-rheumatoid) arthropathy, non-articular rheumatism, and periarticular disorders), and pain associated with degenerative joint diseases (such as arthritis or osteoarthritis-related pain).

[0157] Acute pain: As used in this article, the term "acute pain" refers to sudden or acute pain. Acute pain can be caused, for example, by surgery or acute inflammation.

[0158] Chronic pain: As used in this article, the term "chronic pain" refers to pain that is persistent and typically lasts for more than six months. Chronic pain is not simply a matter of the duration of acute pain. In cases of persistent damage, the functional and structural reorganization of neural circuits in the CNS leads to long-term changes in perception and behavior. This type of pain persists after injury or illness, and pain signals remain active in the nervous system for weeks, months, or years.

[0159] Treatment-resistant pain: As used in this article, the term “treatment-resistant pain” refers to pain that cannot be relieved by conventional analgesics, including anti-inflammatory compounds such as NSAIDs, corticosteroids, and opioid analgesics.

[0160] Nociceptive pain: As used herein, the term "nociceptive pain" refers to pain caused by injury, surgery, or disease affecting tissues (such as arthritis) that stimulates pain receptors. Nociceptive pain also includes chronic pain. Preferred types of nociceptive pain are osteoarthritis-related pain and rheumatoid arthritis pain.

[0161] Inflammatory pain or inflammation-related pain: As used in this article, the term "inflammatory pain or inflammation-related pain" refers to spontaneous hypersensitive pain caused by tissue damage and inflammation (e.g., postoperative pain, trauma, arthritis). Inflammatory pain is a type of nociceptive pain, caused by the activation and sensitization of nociceptors by inflammatory mediators. The pain usually improves as the inflammation subsides.

[0162] Postoperative pain: As used herein, the term "postoperative pain" refers to pain caused or resulting from external trauma (such as cuts, punctures, incisions, tears, or wounds to individual tissues), including pain resulting from all surgical procedures, whether invasive or non-invasive. As used herein, postoperative pain does not include pain that occurs (or originates) without external bodily trauma.

[0163] Pain associated with musculoskeletal disorders: As used in this article, the term “pain associated with musculoskeletal disorders” refers to pain associated with musculoskeletal conditions such as rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, serologically negative (non-rheumatoid) arthropathy, and non-articular rheumatoid diseases.

[0164] Pain associated with degenerative joint disease: As used in this article, the term “pain associated with degenerative joint disease” refers to pain associated with arthritis or osteoarthritis.

[0165] Osteoarthritis (OA)-related pain: As used herein, the term “osteoarthritis (OA)-related pain” or “OA-related pain” refers to a chronic joint disease characterized by loss of articular cartilage, thickening of the joint capsule, and periarticular new bone formation (osteophyte formation), ultimately leading to pain and limb dysfunction. In canines, symptoms of OA are often nonspecific and include: i) impaired mobility, reluctance to move, generalized reduced activity, stiffness, lameness, inability to jump, and gait alterations (such as “bunny hops”); ii) pain during manipulation and behavioral changes (such as signs of aggression or discomfort).

[0166] Chronic refractory OA-related pain: As used in this article, the term "chronic refractory OA-related pain" refers to chronic pain that is unresponsive or only mildly responsive to conventional treatments, including NSAIDs, corticosteroids, and opioids.

[0167] Treatment: As used herein, the terms “treatment,” “treat,” “treated,” or “treating” refer to a therapeutic method. Treatment refers to a method of achieving a beneficial or desired clinical outcome. For the purposes of this invention, a beneficial or desired clinical outcome includes, but is not limited to, one or more of the following: improvement or relief of any aspect of NGF-related conditions, particularly pain selected from the group consisting of: acute, chronic, refractory pain, nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders (such as rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, serologically negative (non-rheumatoid) arthropathy, non-articular rheumatism, and periarticular disorders), and pain associated with degenerative joint diseases (such as arthritis or osteoarthritis-related pain). For the purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, one or more of the following: reduction in the severity or incidence of NGF-related symptoms, relief of one or more symptoms associated with the NGF-related symptoms, preferably pain, including any aspect of pain (such as shortening the duration of NGF-related symptoms (preferably pain), reducing NGF-related symptoms, preferably pain sensitivity or pain sensation). For example, one parameter used to measure the severity, incidence, or reduction of pain associated with OA is the reduction in lameness in treated animals compared to untreated animals.

[0168] Treatment methods: This invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0169] According to other specific examples, the composition in a method for treating NGF-related diseases in canines is an immunogenic composition, and such treatment comprises, preferably, active immunization of the canine against endogenous NGF proteins.

[0170] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the treatment comprises active immunization of the canine against the canine's endogenous NGF protein.

[0171] According to another aspect, the present invention provides a method for treating NGF-related diseases in canines, the method comprising administering a composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0172] According to another aspect, the present invention provides a method for active immunization of canines against NGF-related diseases, the method comprising administering a composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0173] In a preferred specific example, the NGF-related condition is pain, preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint diseases, and / or pain associated with osteoarthritis (OA).

[0174] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related symptom is pain.

[0175] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, preferably wherein such pain is OA-related pain.

[0176] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is nociceptive pain.

[0177] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is inflammatory pain.

[0178] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site; and wherein the NGF-related condition is postoperative pain.

[0179] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is pain associated with musculoskeletal disorders.

[0180] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is pain associated with degenerative joint disease.

[0181] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is osteoarthritis (OA)-related pain.

[0182] For individual NGF-related conditions, pain is preferred, and more preferably pain that is selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, preferably such OA-related pain, which is of an acute, chronic or refractory cause.

[0183] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is an acute NGF-related condition.

[0184] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related symptom is acute pain.

[0185] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is acute pain selected from the group consisting of: acute nociceptive pain, acute inflammatory pain, acute postoperative pain, acute pain associated with musculoskeletal diseases, acute pain associated with degenerative joint diseases and / or acute osteoarthritis (OA)-related pain, preferably wherein such pain is acute pain associated with degenerative joint diseases, and even more preferably acute OA-related pain.

[0186] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is a chronic NGF-related condition.

[0187] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site; and wherein the NGF-related condition is chronic pain.

[0188] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is chronic pain selected from the group consisting of: chronic nociceptive pain, chronic inflammatory pain, chronic postoperative pain, chronic pain associated with musculoskeletal diseases, chronic pain associated with degenerative joint diseases and / or chronic osteoarthritis (OA)-related pain, preferably wherein such chronic pain is chronic pain associated with degenerative joint diseases, and even more preferably chronic OA-related pain.

[0189] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related disease is a refractory NGF-related disease.

[0190] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is intractable pain.

[0191] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is refractory pain selected from the group consisting of: refractory nociceptive pain, refractory inflammatory pain, refractory postoperative pain, refractory pain associated with musculoskeletal diseases, refractory pain associated with degenerative joint diseases and / or refractory osteoarthritis (OA)-related pain, preferably wherein such refractory pain is refractory pain associated with degenerative joint diseases, and even more preferably refractory OA-related pain.

[0192] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related disease is a chronic, refractory NGF-related disease.

[0193] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is chronic refractory pain.

[0194] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site; and wherein the NGF-related condition is chronic refractory pain selected from the group consisting of: chronic refractory nociceptive pain, chronic refractory inflammatory pain, chronic refractory postoperative pain, chronic refractory pain associated with musculoskeletal diseases and / or chronic refractory osteoarthritis (OA)-related pain, preferably wherein such chronic refractory pain is chronic refractory pain associated with degenerative joint diseases, and even more preferably chronic refractory OA-related pain.

[0195] Administration: The compositions used according to the present invention are typically administered in an effective dose to treat NGF-related conditions, preferably pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0196] According to a preferred embodiment of the invention, the composition in the method for treating NGF-related diseases in canines is administered to the canine in an amount of at least 50 µg / dose, preferably 50 to 300 µg / dose, preferably 50 to 275 µg / dose, preferably 50 to 250 µg / dose, preferably 50 to 200 µg / dose, preferably 50 to 175 µg / dose, preferably 50 to 150 µg / dose, preferably 62.5 to 300 µg / dose, preferably 62.5 to 275 µg / dose, preferably 62.5 to 250 µg / dose, preferably 62.5 to 200 µg / dose, preferably 62.5 to 175 µg / dose, preferably 62.5 to 150 µg / dose, preferably 75 to 300 µg / dose. The amount is preferably 75 to 275 µg / dose, preferably 75 to 250 µg / dose, preferably 75 to 200 µg / dose, preferably 75 to 175 µg / dose, preferably 75 to 150 µg / dose, preferably 100 to 300 µg / dose, preferably 100 to 275 µg / dose, preferably 100 to 250 µg / dose, preferably 100 to 200 µg / dose, preferably 100 to 175 µg / dose, preferably 100 to 150 µg / dose.

[0197] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site; and wherein the NGF-related condition is pain, preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain; and wherein the composition is administered to the canine in an amount of at least 50 µg / dose, preferably 50 to 300 µg / dose, preferably 50 to 275 µg / dose, preferably 50 to 250 µg / dose, preferably 50 to 200 µg / dose, preferably 50 to 175 µg / dose, preferably 50 to 150 µg / dose. The amount is preferably 62.5 to 300 µg / dose, preferably 62.5 to 275 µg / dose, preferably 62.5 to 250 µg / dose, preferably 62.5 to 200 µg / dose, preferably 62.5 to 175 µg / dose, preferably 62.5 to 150 µg / dose, preferably 75 to 300 µg / dose, preferably 75 to 275 µg / dose, preferably 75 to 250 µg / dose, preferably 75 to 200 µg / dose, preferably 75 to 175 µg / dose, preferably 75 to 150 µg / dose, preferably 100 to 300 µg / dose, preferably 100 to 275 µg / dose, preferably 100 to 250 µg / dose. µg / dose, preferably 100 to 200 µg / dose, preferably 100 to 175 µg / dose, preferably 100 to 150 µg / dose.

[0198] Application plan: NGF-containing CMV VLP compositions can enhance the neutralization of anti-NGF antibodies after repeated or multiple administrations without inducing any long-term immune tolerance (i.e., not permanently preventing immune tolerance), which would otherwise cause serious side effects and reduce the enhancing effect against endogenous NGF proteins. This temporary breakthrough of immune tolerance after repeated administration, without completely destroying immune tolerance against endogenous NGF proteins, is crucial for managing NGF-related diseases, especially NGF-related pain. Furthermore, a further requirement is that the carrier presenting the NGF antigen (i.e., the CMV VLP) itself does not induce a neutralizing immune response against such a carrier, as this would weaken any enhancing effect after repeated administration. Surprisingly, NGF-containing CMV VLPs have been found to be highly suitable for managing NGF-related conditions in canines, particularly for pain management. The NGF-containing CMV VLP unexpectedly mediated a temporary breakthrough in immune tolerance against endogenous NGF protein and activated and enhanced antibody-mediated inhibition of NGF binding to the TrkA high-affinity receptor, thereby preventing subsequent cascade activation without permanently destroying immune tolerance against endogenous NGF protein.

[0199] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in one or more doses.

[0200] In another aspect, the CMV VLP composition containing NGF is administered in at least two doses, preferably with a time interval of 7 to 21 days between the first and second doses, more preferably 14 to 21 days. Such preferred time intervals between the first and second doses may be 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days, preferably 14, 15, 16, 17, 18, 19, 20, or 21 days.

[0201] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine at least two doses, preferably the time interval between the first dose and the second dose is 7 to 21 days, more preferably 14 to 21 days, more preferably the time interval between the first dose and the second dose is 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, preferably 14, 15, 16, 17, 18, 19, 20 or 21 days.

[0202] In cases of repeated administration, the time interval between the first and second doses is preferably at least 7 days, more preferably at least 14 days, even more preferably 7 to 21 days, and even more preferably 14 to 21 days. Further booster administrations are typically spaced further apart from the previous administration. For example, if a third administration is provided, the time interval between the second and third administrations is generally two to six months. For any further booster administration, the time interval following the previous booster administration (e.g., after the third administration) is preferably at least three months, preferably three to six months.

[0203] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site, and wherein the composition is administered to the canine in repeated doses.

[0204] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in repeated doses, and wherein the time interval between the first dose and the second dose is preferably at least 7 days, preferably at least 14 days, more preferably 7 to 21 days, and more preferably 14 to 21 days.

[0205] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected via at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in repeated doses, wherein the time interval between the first dose and the second dose is preferably at least 7 days, preferably at least 14 days, more preferably 7 to 21 days, more preferably 14 to 21 days, and wherein any subsequent administration is at least two to six months after the previous administration. This time interval between any subsequent administration and the previous administration may be, for example, 2, 3, 4, 5, or 6 months, or any other time interval within 2 to 6 months.

[0206] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in repeated doses, wherein the time interval between the first dose and the second dose is preferably two to three weeks (14 to 21 days), and wherein any subsequent administration is approximately six months after the previous administration.

[0207] For the treatment of acute NGF-related symptoms, preferably acute pain, one or two doses of an NGF-containing CMV VLP composition are generally administered. If two doses are administered, a time interval of one to three weeks, preferably two to three weeks, is typically chosen to obtain the maximum enhancement effect.

[0208] According to another aspect, the present invention provides a composition for a method of treating acute NGF-related symptoms (preferably acute pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in at least two doses, preferably in two doses, preferably with a time interval of 7 to 21 days between the first and second doses, preferably 14 to 21 days (two to three weeks), more preferably with a time interval of 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, more preferably 14, 15, 16, 17, 18, 19, 20 or 21 days. Such acute NGF-related symptoms, preferably such acute pain, are selected from the group consisting of: acute nociceptive pain, acute inflammatory pain, acute postoperative pain, acute pain associated with musculoskeletal diseases, acute pain associated with degenerative joint diseases and / or acute osteoarthritis (OA)-related pain, preferably such acute pain is acute pain associated with degenerative joint diseases, more preferably acute OA-related pain. Preferably, the dosage of the CMV VLP containing NGF is 50 to 300 µg / dose, preferably 50 to 275 µg / dose, preferably 50 to 250 µg / dose, preferably 50 to 200 µg / dose, preferably 50 to 175 µg / dose, preferably 50 to 150 µg / dose, preferably 62.5 to 300 µg / dose, preferably 62.5 to 275 µg / dose, preferably 62.5 to 250 µg / dose, preferably 62.5 to 200 µg / dose, preferably 62.5 to 175 µg / dose, preferably 62.5 to 150 µg / dose, preferably 75 to 300 µg / dose, preferably 75 to 275 µg / dose. µg / dose, preferably 75 to 250 µg / dose, preferably 75 to 200 µg / dose, preferably 75 to 175 µg / dose, preferably 75 to 150 µg / dose, preferably 100 to 300 µg / dose, preferably 100 to 275 µg / dose, preferably 100 to 250 µg / dose, preferably 100 to 200 µg / dose, preferably 100 to 175 µg / dose, preferably 100 to 150 µg / dose.

[0209] For chronic or chronically refractory NGF-related conditions, treatment of chronic pain or chronically refractory pain is preferred, and repeated administration can further enhance the therapeutic effect. If three to four doses are administered, the time interval between the first and second doses is typically 7 to 21 days, preferably 14 to 21 days (two to three weeks), more preferably 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days, and even more preferably 14, 15, 16, 17, 18, 19, 20, or 21 days. Then, the time interval between any subsequent administration and the previous administration is two to six months. For example, the time interval between the second and third administrations can be two months, three months, four months, five months, or six months (or any other time interval in between), and the time interval between any fourth administration and the third administration is three months, four months, five months, or six months (or any other time interval in between). Generally, it is highly desirable to have a six-month interval between the initial administration (which preferably consists of one or two doses) and any subsequent administrations, with each subsequent administration preceding the previous one. Furthermore, the precise time interval for any further booster administrations following one or two initial administrations can be determined by those skilled in the art based on the anti-NGF titers detected in pretreated animals (pretreated canines).

[0210] According to another aspect, the present invention provides a composition for a method of treating chronic NGF-related conditions (preferably chronic pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in repeated doses, wherein repeated administration comprises administering one or two initiating doses or consisting thereof, preferably administering two initiating doses, wherein the time interval between the first initiating dose and the second initiating dose is 7 to 21 days, preferably 14 to 21 days (two to three weeks), more preferably the time interval between the first dose and the second dose is 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, more preferably 14, 15, 16, 17, 18, 19, 20 or 21 days; and wherein the repeated administration comprises or consists of subsequent booster administrations, which are preferably two to six months apart from any previous administration, preferably three, four, five or six months, more preferably four to six months. This type of chronic NGF-related condition, preferably, is a chronic pain selected from the group consisting of: chronic nociceptive pain, chronic inflammatory pain, chronic postoperative pain, chronic pain associated with musculoskeletal disorders, chronic pain associated with degenerative joint diseases, and / or chronic osteoarthritis (OA)-related pain. Preferably, this type of chronic pain is chronic pain associated with degenerative joint diseases, and more preferably, chronic OA-related pain. According to another specific example, this type of chronic NGF-related condition can be chronic refractory pain selected from the group consisting of: chronic refractory nociceptive pain, chronic refractory inflammatory pain, chronic refractory postoperative pain, chronic refractory pain associated with musculoskeletal disorders, and / or chronic refractory osteoarthritis (OA)-related pain. Preferably, this type of chronic pain is chronic refractory pain associated with degenerative joint diseases, and more preferably, chronic refractory OA-related pain.Preferably, the dosage of the (CMV VLP containing NGF) composition is 50 to 300 µg / dose, preferably 50 to 275 µg / dose, preferably 50 to 250 µg / dose, preferably 50 to 200 µg / dose, preferably 50 to 175 µg / dose, preferably 50 to 150 µg / dose, preferably 62.5 to 300 µg / dose, preferably 62.5 to 275 µg / dose, preferably 62.5 to 250 µg / dose, preferably 62.5 to 200 µg / dose, preferably 62.5 to 175 µg / dose, preferably 62.5 to 150 µg / dose, preferably 75 to 300 µg / dose, preferably 75 to 275 µg / dose. µg / dose, preferably 75 to 250 µg / dose, preferably 75 to 200 µg / dose, preferably 75 to 175 µg / dose, preferably 75 to 150 µg / dose, preferably 100 to 300 µg / dose, preferably 100 to 275 µg / dose, preferably 100 to 250 µg / dose, preferably 100 to 200 µg / dose, preferably 100 to 175 µg / dose, preferably 100 to 150 µg / dose.

[0211] According to another aspect, the present invention provides a composition for a method of treating chronic NGF-related conditions (preferably chronic pain) in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected via at least one first attachment site and at least one second attachment site, and wherein the composition is administered to the canine in repeated doses, wherein the repeated administration comprises or consists of: administering two initiating doses, wherein the time interval between the first initiating dose and the second initiating dose is two to three weeks; and administering at least two subsequent booster doses, preferably with the time interval between the second initiating dose and the first booster dose being two to three months; and at least one further booster dose, which is four to six months apart from any previous booster dose. Such chronic NGF-related conditions, preferably such chronic pain, are selected from the group consisting of: chronic nociceptive pain, chronic inflammatory pain, chronic postoperative pain, chronic pain associated with musculoskeletal diseases, chronic pain associated with degenerative joint diseases, and / or chronic osteoarthritis (OA)-related pain, preferably such chronic pain is chronic pain associated with degenerative joint diseases, and more preferably chronic OA-related pain. According to another specific example, such chronic NGF-related conditions may be chronic refractory pain selected from the group consisting of: chronic refractory nociceptive pain, chronic refractory inflammatory pain, chronic refractory postoperative pain, chronic refractory pain associated with musculoskeletal diseases and / or chronic refractory osteoarthritis (OA)-related pain, preferably such chronic pain is chronic refractory pain associated with degenerative joint diseases, and more preferably chronic refractory OA-related pain. Preferably, the dosage of the (CMV VLP containing NGF) composition is 50 to 300 µg / dose, preferably 50 to 275 µg / dose, preferably 50 to 250 µg / dose, preferably 50 to 200 µg / dose, preferably 50 to 175 µg / dose, preferably 50 to 150 µg / dose, preferably 62.5 to 300 µg / dose, preferably 62.5 to 275 µg / dose, preferably 62.5 to 250 µg / dose, preferably 62.5 to 200 µg / dose, preferably 62.5 to 175 µg / dose, preferably 62.5 to 150 µg / dose, preferably 75 to 300 µg / dose, preferably 75 to 275 µg / dose. µg / dose, preferably 75 to 250 µg / dose, preferably 75 to 200 µg / dose, preferably 75 to 175 µg / dose, preferably 75 to 150 µg / dose, preferably 100 to 300 µg / dose, preferably 100 to 275 µg / dose, preferably 100 to 250 µg / dose, preferably 100 to 200 µg / dose, preferably 100 to 175 µg / dose, preferably 100 to 150 µg / dose.

[0212] Application method: Generally, CM VLP compositions containing NGF are administered systemically, preferably subcutaneously, intramuscularly, or transdermally. In a preferred embodiment, the CM VLP composition containing NGF is administered subcutaneously.

[0213] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected via at least one first attachment site and at least one second attachment site, and wherein the composition is administered systemically, preferably subcutaneously, intramuscularly, or transdermally. In a preferred embodiment, the NGF-containing CMV VLP composition is administered subcutaneously.

[0214] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site; and wherein the composition is administered subcutaneously. Preferably, the NGF-related condition is pain, preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint disorders and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint disorders, chronic pain associated with degenerative joint disorders, or even chronic refractory pain associated with degenerative joint disorders, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain. According to another preferred embodiment, the dosage of the systemically, preferably subcutaneously, composition is 50 to 300 µg / dose, preferably 50 to 275 µg / dose, preferably 50 to 250 µg / dose, preferably 50 to 200 µg / dose, preferably 50 to 175 µg / dose, preferably 50 to 150 µg / dose, preferably 62.5 to 300 µg / dose, preferably 62.5 to 275 µg / dose, preferably 62.5 to 250 µg / dose, preferably 62.5 to 200 µg / dose, preferably 62.5 to 175 µg / dose, preferably 62.5 to 150 µg / dose, preferably 75 to 300 µg / dose, preferably 75 to 275 µg / dose. µg / dose, preferably 75 to 250 µg / dose, preferably 75 to 200 µg / dose, preferably 75 to 175 µg / dose, preferably 75 to 150 µg / dose, preferably 100 to 300 µg / dose, preferably 100 to 275 µg / dose, preferably 100 to 250 µg / dose, preferably 100 to 200 µg / dose, preferably 100 to 175 µg / dose, preferably 100 to 150 µg / dose. According to another preferred embodiment, the composition is administered to a canine animal systemically in at least two doses, preferably subcutaneously, with the time interval between the first and second doses preferably being 7 to 21 days, more preferably 14 to 21 days, more preferably 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, and even more preferably 14, 15, 16, 17, 18, 19, 20 and 21 days.In the event of any further application of the composition, such third, optional fourth and any further optional applications comprise or consist of: systemic application, preferably subcutaneous application of the third, optional fourth and optional other doses, preferably at an interval of two to six months from the corresponding previous application, preferably at an interval of four to six months from the corresponding previous application.

[0215] The NGF antigen to be used according to the present invention: In another preferred embodiment, the antigen / NGF antigen is nerve growth factor (NGF), selected from human NGF (hNGF), canine NGF (cNGF), feline NGF (fNGF), equine NGF (eNGF), bovine NGF (bNGF), and suidae NGF (pNGF), preferably selected from canine NGF (cNGF) or feline NGF (fNGF), and more preferably, the antigen is canine NGF (cNGF). In a preferred embodiment, the antigen comprises or preferably consists of the following: an amino acid sequence selected from any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58, or an amino acid sequence having at least 90% or at least 91%, preferably at least 92%, at least 93% or at least 94%, more preferably at least 95%, at least 96% or at least 97%, and even more preferably at least 98% or at least 99% sequence identity with any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, and SEQ ID NO:58. In another embodiment, the NGF antigen comprises at least two consecutive and at most 12 consecutive histidine residues of polyhistidine tag, the tag preferably being located at the C-terminus or N-terminus of the NGF antigen. In another preferred embodiment, the NGF antigen comprises a polyhistidine tag of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 consecutive histidine residues, preferably located at the C-terminus or N-terminus of the NGF antigen. In another preferred embodiment, the NGF antigen comprises a polyhistidine tag of 4, 6, 8, or 10 consecutive histidine residues, preferably located at the C-terminus or N-terminus of the NGF antigen. In another preferred embodiment, the NGF antigen comprises a polyhistidine tag of 4 consecutive histidine residues, preferably located at the C-terminus of the NGF antigen. In another preferred embodiment, the NGF antigen comprises a polyhistidine tag of 4 consecutive histidine residues, preferably located at the N-terminus of the NGF antigen. In yet another preferred embodiment, the NGF antigen comprises a polyhistidine tag of 6 consecutive histidine residues, consisting of SEQ ID NO:34, preferably located at the C-terminus of the NGF antigen. In another specific example, the NGF antigen contains a polyhistidine tag consisting of six consecutive histidine residues of SEQ ID NO:34, which is preferably located at the N-terminus of the NGF antigen.In another specific example, the NGF antigen comprises a polyhistidine tag of 8 consecutive histidine residues, preferably located at the C-terminus of the NGF antigen. In another specific example, the NGF antigen comprises a polyhistidine tag of 8 consecutive histidine residues, preferably located at the N-terminus of the NGF antigen. In yet another specific example, the NGF antigen comprises a polyhistidine tag of 10 consecutive histidine residues, preferably located at either the C-terminus or N-terminus of the NGF antigen.

[0216] In one alternative specific example, the antigen is human NGF. In another specific example, the antigen comprises or preferably consists of: SEQ ID NO:54 or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:54. In another specific example, the antigen comprises SEQ ID NO:54. In yet another, the antigen consists of SEQ ID NO:54.

[0217] In another highly preferred embodiment, the antigen is canine NGF. In another highly preferred embodiment, the antigen comprises or preferably consists of: SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95%, and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:30, SEQ ID NO:31, or SEQ ID NO:33. In another preferred embodiment, the antigen comprises SEQ ID NO:30, SEQ ID NO:31, or SEQ ID NO:33. In another highly preferred embodiment, the antigen comprises or preferably consists of: SEQ ID NO:30, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95%, and even more preferably at least 98%, amino acid sequence identity with SEQ ID NO:30. In another highly preferred embodiment, the antigen comprises SEQ ID NO:30. In another highly preferred embodiment, the antigen is composed of SEQ ID NO:30. In another preferred embodiment, the antigen comprises or preferably comprises: SEQ ID NO:31 or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:31. In another highly preferred embodiment, the antigen comprises SEQ ID NO:31. In another highly preferred embodiment, the antigen is composed of SEQ ID NO:31. In another preferred embodiment, the antigen comprises or preferably comprises: SEQ ID NO:33 or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:33. In another highly preferred embodiment, the antigen comprises SEQ ID NO:33. In another highly preferred embodiment, the antigen is composed of SEQ ID NO:33.

[0218] In another specific example, the antigen comprises or preferably consists of: SEQ ID NO:30, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:30, and wherein the NGF antigen further comprises at least two consecutive and at most 12 consecutive histidine residues, preferably 4, 6, 8 or 10 consecutive histidine residues, more preferably 6 consecutive histidine residues, the tag being composed of SEQ ID NO:34 and preferably located at the C-terminus or N-terminus of the NGF antigen, more preferably at the C-terminus of the NGF antigen. In another specific example, the antigen comprises, or preferably comprises, SEQ ID NO:30, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and more preferably at least 98% amino acid sequence identity with SEQ ID NO:30, and wherein the NGF antigen further comprises a polyhistidine tag of 4, 6, 8, or 10 consecutive histidine residues, preferably 6 consecutive histidine residues, the tag being composed of SEQ ID NO:34 and preferably located at the C-terminus or N-terminus of the NGF antigen, more preferably at the C-terminus of the NGF antigen. In another preferred specific example, the antigen comprises, or preferably comprises, SEQ ID NO:30, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:30, and wherein the NGF antigen further comprises a polyhistidine tag of 6 consecutive histidine residues, the tag being composed of SEQ ID NO:34 and preferably located at the C-terminus or N-terminus of the NGF antigen, more preferably at the C-terminus of the NGF antigen. In another preferred embodiment, the antigen comprises or preferably consists of: SEQ ID NO:30, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95% sequence identity and even more preferably at least 98% amino acid sequence identity with SEQ ID NO:30, wherein the NGF antigen further comprises a polyhistidine tag of 6 consecutive histidine residues, the tag being composed of SEQ ID NO:34 and located at the N-terminus of the NGF antigen. In another specific embodiment, the antigen comprises SEQ ID NO:30, wherein the NGF antigen further comprises at least two consecutive and at most 12 consecutive histidine residues, preferably 4, 6, 8 or 10 consecutive histidine residues, more preferably 6 consecutive histidine residues, the tag being composed of SEQ ID NO:34 and preferably located at the C-terminus or N-terminus of the NGF antigen, more preferably at the C-terminus of the NGF antigen.In another specific example, the antigen comprises SEQ ID NO:30, and wherein the NGF antigen further comprises a polyhistidine tag of six consecutive histidine residues, the tag being composed of SEQ ID NO:34 and preferably located at the C-terminus or N-terminus of the NGF antigen, more preferably at the C-terminus of the NGF antigen. In another specific example, the antigen comprises SEQ ID NO:30, and wherein the NGF antigen further comprises a polyhistidine tag of six consecutive histidine residues, the tag being composed of SEQ ID NO:34 and located at the N-terminus of the NGF antigen. In another very preferred specific example, the antigen comprises SEQ ID NO:30.

[0219] In another specific example, the antigen is feline NGF. In another very preferred specific example, the antigen comprises or preferably consists of the following: SEQ ID NO:55 or an amino acid sequence having at least 90% or at least 91%, preferably at least 92%, at least 93% or at least 94%, more preferably at least 95%, at least 96% or at least 97% sequence identity with SEQ ID NO:55, and even more preferably at least 98% or at least 99% amino acid sequence identity. In another preferred specific example, the antigen comprises SEQ ID NO:55. In another preferred specific example, the antigen is composed of SEQ ID NO:55.

[0220] In another specific example, the antigen is equine NGF. In another specific example, the antigen comprises or preferably consists of the following: SEQ ID NO:56 or an amino acid sequence having at least 90% or at least 91%, preferably at least 92%, at least 93% or at least 94%, more preferably at least 95%, at least 96% or at least 97% sequence identity with SEQ ID NO:56, and even more preferably at least 98% or at least 99% amino acid sequence identity. In another preferred specific example, the antigen comprises SEQ ID NO:56. In another very preferred specific example, the antigen consists of SEQ ID NO:56.

[0221] In another specific example, the antigen is bovine NGF. In another preferred specific example, the antigen comprises or preferably consists of the following: SEQ ID NO:57 or an amino acid sequence having at least 90% or at least 91%, preferably at least 92%, at least 93% or at least 94%, more preferably at least 95%, at least 96% or at least 97% sequence identity with SEQ ID NO:57, and even more preferably at least 98% or at least 99% amino acid sequence identity. In another preferred specific example, the antigen comprises SEQ ID NO:57. In another very preferred specific example, the antigen consists of SEQ ID NO:57.

[0222] In another specific example, the antigen is NGF from the suidae family. In another preferred specific example, the antigen comprises or preferably consists of the following: SEQ ID NO:58 or an amino acid sequence having at least 90% or at least 91%, preferably at least 92%, at least 93% or at least 94%, more preferably at least 95%, at least 96% or at least 97% sequence identity with SEQ ID NO:58, and even more preferably at least 98% or at least 99% amino acid sequence identity. In another preferred specific example, the antigen comprises SEQ ID NO:58. In another very preferred specific example, the antigen consists of SEQ ID NO:58.

[0223] The composition to be used according to the present invention: The cucumber mosaic virus (CMV) virus-like particles (VLPs) applicable to this invention include CMV VLPs known to those skilled in the art, for example from WO2016 / 062720 and WO2020 / 128037, and the references cited therein. Because cucumber mosaic virus has an extremely wide host range, many different strains and isolates of CMV are known, and the sequences of the capsid proteins of said strains and isolates have been determined and are therefore known to those skilled in the art. The sequences of said CMV capsid proteins (CPs) are described in and can be retrieved from known databases such as Genbank, www.dpvweb.net, or www.ncbi.nlm.nih.gov / protein / . Examples are described on pages 12 to 14 of WO2016 / 062720, the disclosure of which is incorporated herein by reference. The CMV VLPs of this invention comprise at least one CMV polypeptide, typically 180 copies of which are used to form a capsid structure. Preferably, the VLP of CMV contains the CMV polypeptide as the main protein component of its capsid structure, and even more preferably as the sole protein component of its capsid structure.

[0224] Preferably, the VLP of CMV contains or is composed of at least one (typically and preferably 180 copies) CMV polypeptide, which contains or is preferably composed of: (i) the amino acid sequence of the CMV shell protein; or (ii) an amino acid sequence having at least 90%, preferably at least 95%, more preferably at least 98%, and even more preferably at least 99% sequence identity with the CMV shell protein.

[0225] In a preferred embodiment, the CMV polypeptide comprises an amino acid sequence of the CMV shell protein or a mutated amino acid sequence, preferably composed of the mutated amino acid sequence, wherein the mutated amino acid sequence exhibits at least 90%, preferably at least 91%, 92%, 93, 94% or 95%, more preferably at least 96%, 97% or 98%, and even more preferably at least 99% sequence identity with the CMV shell protein; wherein preferably, the mutated amino acid sequence differs from the amino acid sequence to be mutated by at least one and at most 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 amino acid residues, and wherein more preferably, the difference is selected from (i) insertion, (ii) deletion, (iii) amino acid exchange, and (iv) any combination of (i) to (iii).

[0226] In a preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39.

[0227] Therefore, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A virus-like particle of cucumber mosaic virus (CMV), wherein the CMV VLP contains at least one first attachment site; and wherein the CMV VLP contains at least one CMV polypeptide, wherein the CMV polypeptide contains a CMV coat protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0228] In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 85% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 92% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 93% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 96% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 97% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 98% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 99% sequence identity with SEQ ID NO:39.

[0229] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and wherein the CMV VLP contains at least one CMV polypeptide, wherein the CMV polypeptide contains the CMV coat protein of SEQ ID NO: 39 or an amino acid sequence having at least 75% sequence identity with SEQ ID NO: 39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0230] In one preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 85% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 92% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 93% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 96% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 97% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 98% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV coat protein of SEQ ID NO:39 or an amino acid sequence having at least 99% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises, preferably consists of, the CMV coat protein of SEQ ID NO:39.

[0231] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and wherein the CMV VLP contains at least one CMV polypeptide, wherein the CMV polypeptide contains the CMV coat protein of SEQ ID NO:39 or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0232] The immunogenicity of the NGF-containing CMV VLP composition described herein can be further enhanced by adding a helper T-cell epitope. Preferably, the helper T-cell epitope is a tetanus toxin-derived helper T-cell epitope or a PADRE sequence. Such helper T-cell epitopes preferably comprise or consist of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Such helper T-cell epitopes can be introduced, preferably into CMV VLPs, particularly into CMV peptides, and more preferably into the coat protein sequence of CMV VLPs. According to a preferred specific example of the CMV VLP containing the helper T-cell epitope, the helper T-cell epitope, preferably a tetanus toxin-derived or PADRE sequence, replaces the N-terminal region of the CMV peptide. Preferably, a helper T cell epitope, preferably derived from tetanus toxin or a PADRE sequence, replaces the N-terminal region of the CMV polypeptide, and preferably, the N-terminal region of the CMV polypeptide corresponds to amino acid residues 2-12 of SEQ ID NO:39.

[0233] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (c) Helper T cell epitopes; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0234] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (c) A helper T cell epitope, wherein the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, preferably wherein the helper T cell epitope comprises or is composed of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0235] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and wherein the CMV VLP contains at least one CMV polypeptide; (b) At least one antigen, wherein the antigen comprises at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (c) A helper T cell epitope, preferably derived from tetanus toxin or a PADRE sequence, preferably comprising or consisting of SEQ ID NO:41 or SEQ ID NO:42, wherein the helper T cell epitope substitutes for the N-terminal region of the CMV polypeptide, preferably amino acids corresponding to amino acid residues 2-12 of SEQ ID NO:39; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0236] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified cucumber mosaic virus (CMV) VLP, wherein the modified CMV VLP comprises at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, consists of, an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the CMV coat protein; and (ii) Helper T cell epitope, wherein preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, amino acids corresponding to amino acid residues 2-12 of SEQ ID NO:39; (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0237] In one preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 75% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 39. In yet another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 92% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 93% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 96% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 97% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 98% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV coat protein of SEQ ID NO:39 or an amino acid sequence having at least 99% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises, preferably consists of, the CMV coat protein of SEQ ID NO:39.

[0238] In another specific embodiment of the present invention, the helper T cell epitope is selected from TT 830-843 (SEQ ID NO:41), PADRE (SEQ ID NO:42), HA 307-319 (SEQ ID NO:43), HBVnc 50-69 (SEQ ID NO:44), CS378-398 (SEQ ID NO:45), MT 17-31 (SEQ ID NO:46), and TT 947-967 (SEQ ID NO:47). In a preferred embodiment, the helper T cell epitope is a helper T cell epitope derived from tetanus toxin or is a PADRE sequence. In a preferred embodiment, the helper T cell epitope is derived from a human vaccine. In a preferred embodiment, the helper T cell epitope is a helper T cell epitope derived from tetanus toxin. In a preferred embodiment, the helper T cell epitope is a PADRE sequence. In a preferred embodiment, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In a very preferred embodiment, the helper T cell epitope consists of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In a very preferred embodiment, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41. In a preferred embodiment, the helper T cell epitope consists of the amino acid sequence of SEQ ID NO:41. In a very preferred embodiment, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:42. In a very preferred embodiment, the helper T cell epitope consists of the amino acid sequence of SEQ ID NO:42.

[0239] Therefore, in another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified cucumber mosaic virus (CMV) VLP, wherein the modified CMV VLP comprises at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: the CMV capsid protein of SEQ ID NO:39, or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 92%, even more preferably at least 95%, even more preferably at least 98% sequence identity with the capsid protein; and (ii) A helper T cell epitope, preferably wherein the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39, more preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, even more preferably, the helper T cell epitope contains the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably it is composed of therein; (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0240] In one preferred embodiment, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, wherein the helper T cell epitope comprises, preferably constitutes, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In another preferred embodiment, the CMV polypeptide is a CMV coat protein or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95%, and even more preferably at least 98% sequence identity with SEQ ID NO:39, wherein the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, wherein the helper T cell epitope comprises, preferably constitutes, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0241] Therefore, in another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified cucumber mosaic virus (CMV) VLP, wherein the modified CMV VLP comprises at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises, preferably, the following: i. A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: the CMV coat protein of SEQ ID NO:39, or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95%, and even more preferably at least 98% sequence identity with SEQ ID NO:39; and ii. A helper T cell epitope, wherein the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, and wherein the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39, and wherein preferably, the helper T cell epitope is derived from tetanus toxin or a PADRE sequence, wherein very preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, and even more preferably, is composed of therefrom; (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected by at least one first attachment site and at least one second attachment site.

[0242] In a preferred embodiment, the modified CMV polypeptide comprises, and preferably consists of, the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 48. In a very preferred embodiment, the modified CMV polypeptide comprises, and preferably consists of, the amino acid sequence of SEQ ID NO: 5.

[0243] Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0244] There are different ways in which CMV VLPs and NGF antigens can be linked. NGF antigens can be linked to CMV VLPs via attachment sites, through peptide bonds, preferably through gene fusion. For example, a nucleotide sequence encoding an NGF antigen can be cloned within an inner frame of a nucleotide sequence encoding a CMV VLP. Alternatively, NGF antigens can be linked to CMV VLPs via chemical coupling between attachment sites, for example, through at least one non-peptide bond.

[0245] In a preferred embodiment, the CMV VLP and the at least one NGF antigen are linked via at least one covalent bond through at least one first attachment site and at least one second attachment site. In another preferred embodiment, the CMV VLP and the at least one NGF antigen are linked solely by covalent bonds through at least one first attachment site and at least one second attachment site.

[0246] In another preferred embodiment, the CMV VLP and the at least one NGF antigen are linked via at least one covalent peptide bond through at least one first attachment site and at least one second attachment site. In another preferred embodiment, the CMV VLP and the at least one NGF antigen are linked only by covalent peptide bonds through at least one first attachment site and at least one second attachment site.

[0247] In another preferred embodiment, the CMV VLP and the at least one NGF antigen are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. In another preferred embodiment, the CMV VLP and the at least one NGF antigen are linked only by covalent non-peptide bonds through at least one first attachment site and at least one second attachment site.

[0248] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected via at least one first attachment site and at least one second attachment site, via at least one non-peptide covalent bond, preferably via fusion.

[0249] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises a modified CMV VLP, and wherein the modified CMV VLP comprises at least one antigenic CMV fusion polypeptide, wherein the at least one antigenic CMV fusion polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (ii) An antigenic NGF polypeptide, wherein the antigenic NGF polypeptide is inserted into the CMV polypeptide, wherein preferably, the antigenic NGF polypeptide is inserted between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39; and (iii) Optionally, a helper T-cell epitope, preferably wherein the helper T-cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T-cell epitope comprises, and preferably consists of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39.

[0250] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises a modified CMV VLP, and wherein the modified CMV VLP comprises at least one antigenic CMV fusion polypeptide, wherein the at least one antigenic CMV fusion polypeptide comprises, preferably, the following: (a) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (b) An antigenic NGF polypeptide, wherein the antigenic NGF polypeptide is inserted into the CMV polypeptide, wherein preferably, the antigenic NGF polypeptide is inserted between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39; and (c) A helper T-cell epitope, preferably derived from tetanus toxin or a PADRE sequence, more preferably comprising, and preferably consisting of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39.

[0251] Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0252] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises a modified CMV VLP, and wherein the modified CMV VLP comprises at least one antigenic CMV fusion polypeptide, wherein the at least one antigenic CMV fusion polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (ii) An antigenic NGF polypeptide, wherein the antigenic NGF polypeptide is inserted into the CMV polypeptide, wherein the antigenic NGF polypeptide is inserted between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39; and (iii) Optionally, a helper T-cell epitope, preferably wherein the helper T-cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T-cell epitope comprises, and preferably consists of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39.

[0253] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises a modified CMV VLP, and wherein the modified CMV VLP comprises at least one antigenic CMV fusion polypeptide, wherein the at least one antigenic CMV fusion polypeptide comprises, preferably, the following: (a) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, even more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (b) An antigenic NGF polypeptide, wherein the antigenic NGF polypeptide is inserted into the CMV polypeptide, wherein the insertion of the antigenic NGF polypeptide is between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39; and (c) A helper T-cell epitope, preferably derived from tetanus toxin or a PADRE sequence, more preferably comprising, and preferably consisting of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39.

[0254] Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0255] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particle (VLP), wherein the CMV VLP contains at least one first attachment site; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected via at least one covalent nonpeptide bond through at least one first attachment site and at least one second attachment site.

[0256] CMV VLPs are typically large molecular assemblies composed of viral capsid proteins, typically containing 180 protein subunits per VLP. Typically and preferably, the interaction of these subunits leads to the formation of VLPs with inherently repeating organization, thereby allowing the presentation of multiple copies of the NGF antigen. Such capsid proteins are, for example, those encoded by the amino acid sequence SEQ ID NO:39 or having at least 75% sequence identity with SEQ ID NO:39.

[0257] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one CMV polypeptide, wherein the at least one CMV polypeptide comprises a CMV polypeptide, preferably composed of a CMV polypeptide, wherein the CMV polypeptide contains a CMV coat protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the NGF antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0258] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (ii) Optionally, helper T cell epitopes; and (b) wherein the antigen contains at least one second attachment site, and wherein the NGF antigen is a nerve growth factor (NGF) antigen; and (a) and (b) are connected via at least one covalent nonpeptide bond through at least one first attachment site and at least one second attachment site.

[0259] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (c) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (ii) Helper T cell epitopes; and (d) wherein the antigen contains at least one second attachment site, and wherein the NGF antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0260] In one preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 75% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 39. In yet another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO: 39 or an amino acid sequence having at least 92% sequence identity with SEQ ID NO: 39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 93% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 96% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 97% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV shell protein of SEQ ID NO:39 or an amino acid sequence having at least 98% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises the CMV coat protein of SEQ ID NO:39 or an amino acid sequence having at least 99% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises, preferably consists of, the CMV coat protein of SEQ ID NO:39.In a preferred embodiment, the helper T cell epitope is selected from TT 830-843 (SEQ ID NO:41), PADRE (SEQ ID NO:42), HA 307-319 (SEQ ID NO:43), HBVnc 50-69 (SEQ ID NO:44), CS 378-398 (SEQ ID NO:45), MT 17-31 (SEQ ID NO:46), and TT 947-967 (SEQ ID NO:47). In a preferred embodiment, the helper T cell epitope is a helper T cell epitope derived from tetanus toxin or is a PADRE sequence. In a preferred embodiment, the helper T cell epitope is derived from a human vaccine. In a preferred embodiment, the helper T cell epitope is a helper T cell epitope derived from tetanus toxin. In a preferred embodiment, the helper T cell epitope is a PADRE sequence. In a preferred embodiment, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In a very preferred embodiment, the helper T cell epitope consists of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In a very preferred embodiment, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41. In a preferred embodiment, the helper T cell epitope consists of the amino acid sequence of SEQ ID NO:41. In a very preferred embodiment, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:42. In a very preferred embodiment, the helper T cell epitope consists of the amino acid sequence of SEQ ID NO:42. In a very preferred embodiment, the modified CMV polypeptide comprises the amino acid sequence of SEQ ID NO:5 or SEQ ID NO:48, preferably. In a very preferred embodiment, the modified CMV polypeptide comprises the amino acid sequence of SEQ ID NO:5, preferably.

[0261] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one modified CMV polypeptide, wherein the at least one modified CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably, the following: a CMV capsid protein or an amino acid sequence having at least 75%, preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 92%, more preferably at least 95%, more preferably at least 98% sequence identity with the capsid protein of SEQ ID NO:39; and (ii) A helper T-cell epitope, preferably derived from tetanus toxin or a PADRE sequence, more preferably comprising, and preferably consisting of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39; and (b) wherein the antigen contains at least one second attachment site, and wherein the NGF antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0262] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising a continuous segment of negatively charged amino acids, preferably composed thereof, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, preferably wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39. (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0263] Therefore, in another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39. (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0264] In a preferred embodiment, the chimeric CMV polypeptide further comprises a helper T-cell epitope, preferably derived from tetanus toxin or a PADRE sequence, preferably comprising the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably composed of therein, and preferably wherein the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, preferably the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39. In another very preferred embodiment, the CMV polypeptide is a CMV coat protein or an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 95%, and more preferably at least 98% sequence identity with SEQ ID NO:39.

[0265] Therefore, in another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and (iii) A helper T cell epitope, preferably wherein the helper T cell epitope replaces the N-terminal region of the CMV polypeptide; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence; preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably, is composed of therein. Preferably, the NGF-related symptom is pain; more preferably, it is pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain; more preferably, it is acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases; more preferably, it is acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0266] In another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39. (iii) A helper T cell epitope, preferably wherein the helper T cell epitope replaces the N-terminal region of the CMV polypeptide; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably, it is composed of thereto. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain. According to another aspect, the present invention provides a composition in a method for treating NGF-related symptoms in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and (iii) A helper T-cell epitope, preferably derived from tetanus toxin or a PADRE sequence, more preferably comprising, and preferably consisting of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0267] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39. (iii) A helper T-cell epitope, preferably derived from tetanus toxin or a PADRE sequence, more preferably comprising, and preferably consisting of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. Preferably, the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide, and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0268] In another highly preferred embodiment, the continuous segment of negatively charged amino acids comprises SEQ ID NO:1 or SEQ ID NO:2, preferably consisting of thereto.

[0269] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39, wherein preferably, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the continuous segment of negatively charged amino acids comprises, preferably, SEQ ID NO:1 or SEQ ID NO:2. (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0270] Therefore, according to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39, wherein preferably, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising a continuous segment of negatively charged amino acids, preferably composed therefrom, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39, and wherein the continuous segment of negatively charged amino acids comprises SEQ ID NO:1 or SEQ ID NO:2, preferably composed therefrom; (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0271] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39, wherein preferably, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39; (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the continuous segment of negatively charged amino acids comprises, preferably, SEQ ID NO:1 or SEQ ID NO:2; and (iii) A helper T-cell epitope, preferably wherein the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide and / or preferably wherein the helper T-cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T-cell epitope comprises, and preferably consists of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, which corresponds to amino acids 2-12 of SEQ ID NO: 39. Preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence. Preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO: 41 or SEQ ID NO: 42, more preferably it is composed of. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0272] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39, wherein preferably, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39; (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39, and wherein the continuous segment of negatively charged amino acids comprises, preferably, SEQ ID NO:1 or SEQ ID NO:2; and (iii) A helper T-cell epitope, preferably wherein the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide and / or preferably wherein the helper T-cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T-cell epitope comprises, and preferably consists of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, which corresponds to amino acids 2-12 of SEQ ID NO:39. Preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence. Preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably, it is composed of the above. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0273] In a preferred embodiment, the polypeptide comprising the continuous segment of negatively charged amino acids further comprises a first amino acid linker and a second amino acid linker, wherein the first amino acid linker is located at the N-terminus of the continuous segment of negatively charged amino acids, and the second amino acid linker is located at the C-terminus of the continuous segment of negatively charged amino acids, and wherein the first amino acid linker and the second amino acid linker are independently selected from the group consisting of: (a.) polyglycine linkers (G-linkers) having an amino acid sequence (Gly) of length n=2 to 10. n (b.) A glycine-serine linker (GS-linker) comprising at least one glycine and at least one serine, wherein preferably, the GS-linker has (GS) r (G s S) t (GS) uThe amino acid sequence, wherein r = 0 or 1, s = 1-5, t = 1-5 and u = 0 or 1; and (c.) an amino acid linker (GS*-linker) comprising at least one Gly, at least one Ser, and at least one amino acid selected from Thr, Ala, Lys, and Cys. In a preferred embodiment, the polypeptide comprising the segment of the consecutive negatively charged amino acids further comprises a first amino acid linker and a second amino acid linker, wherein the first amino acid linker is located at the N-terminus of the segment of the consecutive negatively charged amino acids, and the second amino acid linker is located at the C-terminus of the segment of the consecutive negatively charged amino acids, and wherein the first amino acid linker and the second amino acid linker are independently selected from: a glycine-serine linker (GS-linker) comprising at least one glycine and at least one serine, wherein the GS-linker has (GS) r (G s S) t (GS) u The amino acid sequence, wherein r=0 or 1, s=1-5, t=1-5 and u=0 or 1; and an amino acid linker (GS*-linker) comprising at least one Gly, at least one Ser and at least one amino acid selected from Thr, Ala, Lys and Cys. In another highly preferred embodiment, the first amino acid linker comprises SEQ ID NO:8, preferably. In another highly preferred embodiment, the second amino acid linker comprises SEQ ID NO:4 or SEQ ID NO:9, preferably. In another highly preferred embodiment, the polypeptide comprises SEQ ID NO:49, SEQ ID NO:50 or SEQ ID NO:51. In another highly preferred embodiment, the polypeptide is composed of SEQ ID NO:49, SEQ ID NO:50 or SEQ ID NO:51. In another highly preferred embodiment, the polypeptide comprising a continuous segment of negatively charged amino acids, preferably composed of such segments, is inserted between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39.

[0274] Therefore, in another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39, wherein preferably, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39, and wherein the polypeptide comprises, preferably, SEQ ID NO:49, SEQ ID NO:50, or SEQ ID NO:51, and wherein preferably, the polypeptide is inserted between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0275] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39, wherein preferably, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39; (ii) A polypeptide comprising, preferably, a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is preferably inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39, and wherein the polypeptide comprises, preferably comprises, SEQ ID NO:49, SEQ ID NO:50 or SEQ ID NO:51; and wherein preferably, the polypeptide is inserted between amino acid residues in the CMV polypeptide corresponding to amino acid residues at positions 84 and 85 of SEQ ID NO:39; (iii) A helper T-cell epitope, preferably wherein the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide and / or preferably wherein the helper T-cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T-cell epitope comprises, and preferably consists of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, corresponding to amino acids 2-12 of SEQ ID NO:39. Preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence; preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably, it consists of such amino acids.

[0276] In another highly preferred embodiment, the CMV polypeptide comprises, preferably consists of, the amino acid sequence of SEQ ID NO:5, SEQ ID NO:39, or SEQ ID NO:48, wherein the polypeptide comprising the segment containing the consecutive negatively charged amino acids is inserted between amino acid residues at positions 88 and 89 of SEQ ID NO:5, between amino acid residues at positions 84 and 85 of SEQ ID NO:39, or between amino acid residues at positions 86 and 87 of SEQ ID NO:48. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0277] Therefore, in another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; (ii) A polypeptide comprising a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39; and The CMV polypeptide comprises, preferably, the amino acid sequence of SEQ ID NO:5, SEQ ID NO:39, or SEQ ID NO:48, and the polypeptide comprising the continuous negatively charged amino acid segment is inserted between amino acid residues at positions 88 and 89 of SEQ ID NO:5, between amino acid residues at positions 84 and 85 of SEQ ID NO:39, or between amino acid residues at positions 86 and 87 of SEQ ID NO:48; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the NGF-related condition is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0278] According to another aspect, the present invention provides a composition for a method of treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP contains at least one first attachment site, and wherein the modified CMV VLP contains at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) A CMV polypeptide, wherein the CMV polypeptide comprises, preferably constitutes, the amino acid sequence of SEQ ID NO:5, SEQ ID NO:39 or SEQ ID NO:48; (ii) A polypeptide comprising a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, and the polypeptide comprising the continuous segment of negatively charged amino acids is inserted between amino acid residues at positions 88 and 89 of SEQ ID NO: 5, between amino acid residues at positions 84 and 85 of SEQ ID NO: 39, or between amino acid residues at positions 86 and 87 of SEQ ID NO: 48; and (iii) A helper T-cell epitope, preferably wherein the helper T-cell epitope replaces the N-terminal region of the CMV polypeptide and / or preferably wherein the helper T-cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T-cell epitope comprises, and preferably consists of, the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42; and (b) At least one antigen, wherein the antigen contains at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are linked via at least one covalent non-peptide bond through at least one first attachment site and at least one second attachment site. Preferably, the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, which corresponds to amino acids 2-12 of SEQ ID NO:39. Preferably, the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence. Preferably, the helper T cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, more preferably, it is composed of the above. Preferably, the NGF-related symptom is pain, more preferably pain selected from the group consisting of: nociceptive pain, inflammatory pain, postoperative pain, pain associated with musculoskeletal diseases, pain associated with degenerative joint diseases and / or osteoarthritis (OA)-related pain, more preferably acute pain associated with degenerative joint diseases, chronic pain associated with degenerative joint diseases, or even chronic refractory pain associated with degenerative joint diseases, more preferably acute OA-related pain, chronic OA-related pain, or even chronic refractory OA-related pain.

[0279] The specific examples, preferred examples, and highly preferred examples described and disclosed herein shall apply to all aspects and other specific examples, preferred examples, and / or highly preferred examples, whether or not specifically mentioned again or whether or not repetition is avoided for the sake of brevity.

[0280] In a preferred embodiment, the CMV polypeptide comprises an amino acid sequence of the CMV shell protein or a mutant amino acid sequence, preferably composed of the like, wherein the mutant amino acid sequence exhibits at least 90%, preferably at least 91%, 92%, 93, 94% or 95%, more preferably at least 96%, 97% or 98%, and more preferably at least 99% sequence identity with the CMV shell protein; wherein preferably, the mutant amino acid sequence differs from the amino acid sequence to be mutated by at least one and at most 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 amino acid residues, and wherein more preferably, the difference is selected from (i) insertion, (ii) deletion, (iii) amino acid exchange, and (iv) any combination of (i) to (iii).

[0281] In one preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 85% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39.

[0282] In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 92% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 93% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 96% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 97% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 98% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 99% sequence identity with SEQ ID NO:39.

[0283] In one preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 85% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 92% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 93% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 96% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 97% sequence identity with SEQ ID NO:39. In another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 98% sequence identity with SEQ ID NO:39. In yet another preferred embodiment, the CMV polypeptide is composed of a CMV shell protein or an amino acid sequence having at least 99% sequence identity with SEQ ID NO:39.

[0284] In a preferred embodiment, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 75%, preferably 85%, sequence identity with SEQ ID NO:39. In a preferred embodiment, the CMV polypeptide is a CMV shell protein or an amino acid sequence having at least 90%, preferably 95%, sequence identity with SEQ ID NO:39. In a preferred embodiment, the CMV polypeptide is a CMV shell protein having SEQ ID NO:39. In a preferred embodiment, the CMV shell protein comprises SEQ ID NO:39. In a preferred embodiment, the CMV shell protein is composed of SEQ ID NO:39. In a preferred embodiment, the CMV polypeptide comprises a CMV shell protein. In a preferred embodiment, the CMV polypeptide is composed of a CMV shell protein. In a preferred embodiment, the CMV polypeptide comprises a CMV shell protein, wherein the CMV shell protein comprises SEQ ID NO:39. In a preferred embodiment, the CMV polypeptide comprises a CMV shell protein, wherein the CMV shell protein is composed of SEQ ID NO:39. In a preferred embodiment, the CMV polypeptide is composed of a CMV coat protein, wherein the CMV coat protein is composed of SEQ ID NO:39.

[0285] In one preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 75% sequence identity with SEQ ID NO:40. In another preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 80% sequence identity with SEQ ID NO:40. In yet another preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 85% sequence identity with SEQ ID NO:40. In yet another preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 90% sequence identity with SEQ ID NO:40. In yet another preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 95% sequence identity with SEQ ID NO:40. In a preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 98% sequence identity with SEQ ID NO:40. In another preferred embodiment, the CMV polypeptide comprises either SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 99% sequence identity with SEQ ID NO:40.

[0286] In a preferred embodiment, the CMV polypeptide comprises or preferably consists of: (i) an amino acid sequence of a CMV shell protein, wherein the amino acid sequence comprises or preferably consists of SEQ ID NO:39; or (ii) an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39; and wherein the amino sequence as defined in (i) or (ii) comprises SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 90% sequence identity with SEQ ID NO:40. In a preferred embodiment, the CMV polypeptide comprises or preferably consists of: (i) an amino acid sequence of a CMV shell protein, wherein the amino acid sequence comprises or preferably consists of SEQ ID NO:39; or (ii) an amino acid sequence having at least 95% sequence identity with SEQ ID NO:39; and wherein the amino sequence as defined in (i) or (ii) comprises SEQ ID NO:40 or an amino acid sequence region, wherein the amino acid sequence region has at least 95% sequence identity with SEQ ID NO:40. In a preferred embodiment, the CMV polypeptide comprises or preferably consists of: (i) an amino acid sequence of a CMV shell protein, wherein the amino acid sequence comprises or preferably consists of SEQ ID NO:39; or (ii) an amino acid sequence having at least 90% sequence identity with SEQ ID NO:39; and wherein the amino sequence as defined in (i) or (ii) comprises SEQ ID NO:40.

[0287] In a preferred embodiment, the number of amino acid substitutions in the N-terminal region is equal to or less than the number of amino acids constituting the helper T cell epitope. In a preferred embodiment, the substituted N-terminal region of the CMV polypeptide consists of 5 to 15 consecutive amino acids. In a preferred embodiment, the substituted N-terminal region of the CMV polypeptide consists of 9 to 14 consecutive amino acids. In a preferred embodiment, the substituted N-terminal region of the CMV polypeptide consists of 11 to 13 consecutive amino acids. In a preferred embodiment, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39. In a preferred embodiment, the N-terminal region of the CMV polypeptide contains amino acids 2-12 of SEQ ID NO:39. In a preferred embodiment, the N-terminal region of the CMV polypeptide is composed of amino acids 2-12 of SEQ ID NO:39. In a preferred embodiment, the helper T cell epitope consists of up to 20 amino acids.

[0288] In a preferred embodiment of the present invention, the Th cell epitope is selected from TT 830-843 (SEQ ID NO:41), PADRE (SEQ ID NO:42), HA 307-319 (SEQ ID NO:43), HBVnc 50-69 (SEQ ID NO:44), CS378-398 (SEQ ID NO:45), MT 17-31 (SEQ ID NO:46), and TT 947-967 (SEQ ID NO:47). In a preferred embodiment, the Th cell epitope is a Th cell epitope derived from tetanus toxin or is a PADRE sequence. In a preferred embodiment, the helper T cell epitope is derived from a human vaccine. In a preferred embodiment, the Th cell epitope is a Th cell epitope derived from tetanus toxin. In a preferred embodiment, the Th cell epitope is a PADRE sequence. In a preferred embodiment, the Th cell epitope comprises the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In a highly preferred embodiment, the Th cell epitope consists of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42. In a highly preferred embodiment, the Th cell epitope comprises the amino acid sequence of SEQ ID NO:41. In a preferred embodiment, the Th cell epitope consists of the amino acid sequence of SEQ ID NO:41. In a highly preferred embodiment, the Th cell epitope comprises the amino acid sequence of SEQ ID NO:42. In a highly preferred embodiment, the Th cell epitope consists of the amino acid sequence of SEQ ID NO:42.

[0289] In a preferred embodiment, the CMV polypeptide comprises or preferably consists of the amino acid sequence of the CMV coat protein, wherein the amino acid sequence comprises or preferably consists of the amino acid sequence of SEQ ID NO:39 or having at least 95% sequence identity with SEQ ID NO:39; and wherein the amino sequence comprises SEQ ID NO:40; and wherein the helper T cell epitope replaces the N-terminal region of the CMV polypeptide; and wherein the substituted N-terminal region of the CMV polypeptide consists of 11 to 13 consecutive amino acids, preferably 11 consecutive amino acids; and more preferably, the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:39. In a preferred embodiment, the chimeric CMV polypeptide comprises or preferably consists of the amino acid sequence of SEQ ID NO:5, wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39. In another preferred embodiment, the chimeric CMV polypeptide comprises the amino acid sequence of SEQ ID NO:48, wherein the polypeptide is inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between positions 75 and 85 of SEQ ID NO:39.

[0290] In one preferred embodiment, the length of the consecutive negatively charged amino acid segment is 3 and less than 12 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 3 to 10 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 3, 4, 5, 6, 7, 8, 9, or 10 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 3 to 9 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 3 to 8 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 4 to 9 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 4 to 8 amino acids. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 4, 5, 6, 7, or 8. In another preferred embodiment, the length of the consecutive negatively charged amino acid segment is 4 or 8 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 3 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 5 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 6 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 7 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 8 amino acids. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 9 amino acids.

[0291] In another preferred embodiment, the consecutive segments of negatively charged amino acids are independently selected from aspartic acid or glutamic acid, wherein the aspartic acid or glutamic acid is independently selected from its L-configuration or its D-configuration in each instance. In another preferred embodiment, the consecutive segments of negatively charged amino acids comprise at least one aspartic acid in either the L-configuration or D-configuration. In another preferred embodiment, the consecutive segments of negatively charged amino acids comprise at least one aspartic acid in the L-configuration. In another preferred embodiment, the consecutive segments of negatively charged amino acids comprise at least one aspartic acid in the D-configuration. In another preferred embodiment, the consecutive segments of negatively charged amino acids comprise at least one glutamic acid in the L-configuration. In another preferred embodiment, the consecutive segments of negatively charged amino acids comprise at least one glutamic acid in the D-configuration.

[0292] In another preferred embodiment, the continuous segment of negatively charged amino acids comprises at least one L-configured aspartic acid and at least one L-configured glutamic acid. In another preferred embodiment, the continuous segment of negatively charged amino acids consists entirely of L-configured aspartic acid and glutamic acid. In yet another preferred embodiment, the continuous segment of negatively charged amino acids consists entirely of L-configured aspartic acid or glutamic acid.

[0293] In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least one aspartic acid or at least one glutamic acid. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least two aspartic acid segments or at least two glutamic acid segments. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least three aspartic acid segments or at least three glutamic acid segments. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least four aspartic acid segments or at least four glutamic acid segments. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least four glutamic acid segments. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least five glutamic acid segments. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least six glutamic acid segments. In another preferred embodiment, the consecutive negatively charged amino acid segments comprise at least seven glutamic acid segments. In another preferred embodiment, the continuous segment of negatively charged amino acids comprises at least eight glutamic acids. In another preferred embodiment, the continuous segment of negatively charged amino acids consists only of aspartic acid. In another very preferred embodiment, the continuous segment of negatively charged amino acids consists only of glutamic acid.

[0294] In another preferred embodiment, the continuous negatively charged amino acid segment comprises at least one aspartic acid or at least one glutamic acid, wherein the at least one aspartic acid or the at least one glutamic acid is in the L-configuration. In another preferred embodiment, the continuous negatively charged amino acid segment comprises at least two aspartic acids or at least two glutamic acids, wherein the at least two aspartic acids or at least two glutamic acids are in the L-configuration. In another preferred embodiment, the continuous negatively charged amino acid segment comprises at least four aspartic acids or at least four glutamic acids, wherein the at least four aspartic acids or at least four glutamic acids are in the L-configuration. In another preferred embodiment, the continuous negatively charged amino acid segment comprises at least four aspartic acids, wherein the at least four aspartic acids are in the L-configuration. In another preferred embodiment, the continuous segment of negatively charged amino acids comprises at least four glutamic acids, wherein the at least four glutamic acids are in the L-configuration. In another preferred embodiment, the continuous segment of negatively charged amino acids comprises at least five glutamic acids, wherein the at least five glutamic acids are in the L-configuration. In another preferred embodiment, the continuous segment of negatively charged amino acids comprises at least six glutamic acids, wherein the at least six glutamic acids are in the L-configuration. In another preferred embodiment, the continuous segment of negatively charged amino acids comprises at least seven glutamic acids, wherein the at least seven glutamic acids are in the L-configuration. In another preferred embodiment, the continuous segment of negatively charged amino acids comprises only aspartic acid, wherein the aspartic acid is in the L-configuration. In another very preferred embodiment, the continuous segment of negatively charged amino acids comprises only glutamic acid, wherein the glutamic acid is in the L-configuration.

[0295] In one preferred embodiment, the length of the continuous negatively charged amino acid segment is 3 to 10 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 3 to 9 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 3 to 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 to 9 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 to 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4, 5, 6, 7, or 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 to 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4, 5, 6, 7, or 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 or 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 3 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 5 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 6 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 7 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid.In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 9 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid.

[0296] In one preferred embodiment, the length of the consecutive negatively charged amino acid segments is 3 to 10 amino acids, wherein the consecutive negatively charged amino acid segments consist only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the consecutive negatively charged amino acid segments is 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, wherein the consecutive negatively charged amino acid segments consist only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the consecutive negatively charged amino acid segments is 3 to 9 amino acids, wherein the consecutive negatively charged amino acid segments consist only of glutamic acid, wherein the glutamic acid is in the L-configuration. In yet another preferred embodiment, the length of the consecutive negatively charged amino acid segments is 3 to 8 amino acids, wherein the consecutive negatively charged amino acid segments consist only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 to 9 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 to 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4, 5, 6, 7, or 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 to 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4, 5, 6, 7, or 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 or 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 3 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 4 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 5 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration.In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 6 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 7 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 8 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration. In another preferred embodiment, the length of the continuous negatively charged amino acid segment is 9 amino acids, wherein the continuous negatively charged amino acid segment consists only of glutamic acid, wherein the glutamic acid is in the L-configuration.

[0297] In another highly preferred embodiment, the continuous segment of negatively charged amino acids comprises SEQ ID NO:1 or SEQ ID NO:2. In another highly preferred embodiment, the continuous segment of negatively charged amino acids is composed of SEQ ID NO:1 or SEQ ID NO:2. In another highly preferred embodiment, the continuous segment of negatively charged amino acids comprises SEQ ID NO:1. In another highly preferred embodiment, the continuous segment of negatively charged amino acids is composed of SEQ ID NO:1. In another highly preferred embodiment, the continuous segment of negatively charged amino acids comprises SEQ ID NO:2. In another highly preferred embodiment, the continuous segment of negatively charged amino acids is composed of SEQ ID NO:2.

[0298] In a preferred embodiment, the polypeptide comprising the continuous segment of negatively charged amino acids further comprises a first amino acid linker, wherein the first amino acid linker is located at the N-terminus or C-terminus of the continuous segment of negatively charged amino acids. In a preferred embodiment, the polypeptide further comprises a first amino acid linker, wherein the first amino acid linker is located at the N-terminus of the continuous segment of negatively charged amino acids. In a preferred embodiment, the polypeptide comprising the continuous segment of negatively charged amino acids further comprises a first amino acid linker, wherein the first amino acid linker is located at the C-terminus of the continuous segment of negatively charged amino acids. In a preferred embodiment, the polypeptide comprising the continuous segment of negatively charged amino acids further comprises a second amino acid linker. In a preferred embodiment, the polypeptide comprising the continuous segment of negatively charged amino acids further comprises both a first amino acid linker and a second amino acid linker, wherein the first amino acid linker is located at the N-terminus of the continuous segment of negatively charged amino acids, and the second amino acid linker is located at the C-terminus of the continuous segment of negatively charged amino acids.

[0299] In a preferred embodiment, the length of the first amino acid linker is at most 30 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 20, 19, 18, 17, or 16 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 15 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 14 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 13 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 12 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 11 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 10 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 9 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 8 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 7 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 6 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 5 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 4 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 3 amino acids. In a preferred embodiment, the length of the first amino acid linker is at most 2 amino acids. In a preferred embodiment, the first amino acid linker consists of one amino acid. In a preferred embodiment, the length of the second amino acid linker is at most 30 amino acids. In a preferred embodiment, the length of the second amino acid linker is at most 20, 19, 18, 17, or 16 amino acids. In a preferred embodiment, the length of the second amino acid linker is at most 15 amino acids. In a preferred embodiment, the length of the second amino acid linker is at most 14 amino acids. In a preferred embodiment, the length of the second amino acid linker is at most 13 amino acids. In a preferred embodiment, the length of the second amino acid linker is at most 12 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 11 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 10 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 9 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 8 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 7 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 6 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 5 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 4 amino acids. In one preferred embodiment, the length of the second amino acid linker is at most 3 amino acids...

Claims

1. A composition in a method for treating NGF-related diseases in canines, the method comprising administering the composition to the canine, wherein the composition comprises: (a) Cucumber mosaic virus (CMV) virus-like particles (VLPs), wherein the CMV VLPs contain at least one first attachment site; and (b) At least one antigen, wherein the antigen comprises at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site.

2. The composition for the purpose of claim 1, wherein the NGF-related condition is pain.

3. The composition for the use of claim 2, wherein the pain is selected from the group consisting of: nociceptive pain, inflammation-related pain, postoperative pain, pain associated with musculoskeletal disorders, pain associated with degenerative joint diseases, and / or osteoarthritis (OA)-related pain.

4. The composition for the purpose of claim 2 or 3, wherein the pain is pain associated with degenerative joint disease, preferably OA-related pain.

5. The composition for the use of claims 2 to 4, wherein the pain is acute pain associated with degenerative joint disease, preferably acute OA-related pain.

6. The composition for the use of claims 2 to 4, wherein the pain is chronic pain associated with degenerative joint disease, preferably chronic OA-related pain.

7. The composition for the use of claims 2 to 4, wherein the pain is refractory pain associated with degenerative joint disease, preferably refractory OA-related pain.

8. The composition for the use of claim 6 or 7, wherein the chronic OA-related pain is chronic refractory pain associated with degenerative joint disease, preferably chronic refractory OA-related pain.

9. The composition for the stated purpose as claimed in claims 1 to 8, wherein the composition is administered to the canine in one or more doses.

10. The composition for the said use as claimed in claim 9, wherein the composition is administered to the canine in at least two doses, wherein the time interval between the first dose and the second dose is at least 7 days, preferably at least 14 days.

11. The composition for the stated use as claimed in claim 9 or 10, wherein the composition is administered to the canine in at least two doses, preferably two doses, wherein the time interval between the first dose and the second dose is 7 to 21 days, preferably 14 to 21 days.

12. The composition for the said use as claimed in claims 9 to 11, wherein the composition is administered to the canine in at least three doses, preferably four doses, wherein the time interval between the first dose and the second dose is one to three weeks, preferably two to three weeks, the time interval between the second dose and the third dose is two to six months, and the time interval between any other dose and the previous dose is three to six months.

13. The composition for the stated use according to claims 1 to 12, wherein the composition is administered to the canine in an amount of 50 to 300 µg / dose.

14. The composition for the stated purpose as claimed in claims 1 to 13, wherein the composition is administered subcutaneously, intramuscularly, or percutaneously, preferably subcutaneously, to the canine.

15. The composition for the said use according to claims 1 to 14, wherein (a) and (b) are linked via at least one covalent non-peptide bond through the at least one first attachment site and the at least one second attachment site.

16. The composition for the said use according to claims 1 to 14, wherein the VLP comprises an antigenic VLP fusion polypeptide, and wherein (a) and (b) are fused together via at least one covalent peptide bond through the at least one first attachment site and the at least one second attachment site.

17. The composition for the use of claims 1 to 16, wherein the CMV VLP comprising the at least one first attachment site comprises a CMV polypeptide, preferably composed of a CMV polypeptide, wherein the CMV polypeptide comprises a CMV shell protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:

39.

18. The composition for the said use as claimed in claims 1 to 17, wherein the composition comprises: (a) A modified CMV VLP, wherein the modified CMV VLP comprises at least one first attachment site, and wherein the modified CMV VLP comprises at least one chimeric CMV polypeptide, wherein the at least one chimeric CMV polypeptide comprises, preferably, the following: (i) a CMV polypeptide, wherein the CMV polypeptide comprises a CMV coat protein or an amino acid sequence having at least 75% sequence identity with SEQ ID NO:39; and (ii) A polypeptide comprising a continuous segment of negatively charged amino acids, preferably composed of a continuous segment of negatively charged amino acids, wherein the negatively charged amino acids are independently selected from aspartic acid or glutamic acid, wherein the polypeptide is preferably inserted between any amino acid residues in the CMV polypeptide corresponding to any amino acid residue between position 75 and position 85 of SEQ ID NO:

39. (b) At least one antigen, wherein the antigen comprises at least one second attachment site, and wherein the antigen is a nerve growth factor (NGF) antigen; and Wherein (a) and (b) are connected through the at least one first attachment site and the at least one second attachment site.

19. The composition for the said use according to claims 1 to 18, wherein the CMV VLP further comprises a helper T cell epitope.

20. The composition for the use of claim 19, wherein the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence.

21. The composition for the use of claim 19 or 20, wherein the CMV VLP further comprises a helper T cell epitope, preferably wherein the helper T cell epitope is derived from tetanus toxin or is a PADRE sequence, more preferably wherein the helper T cell epitope is tetanus toxin, the tetanus toxin comprising or consisting of the amino acid sequence of SEQ ID NO:41 or SEQ ID NO:42, and wherein the helper T cell epitope replaces the N-terminal region of the CMV polypeptide, wherein the N-terminal region of the CMV polypeptide corresponds to amino acids 2-12 of SEQ ID NO:

39.

22. The composition for the said use according to claims 18 to 21, wherein the polypeptide comprising the continuous negatively charged amino acid segments further comprises a first amino acid linker and a second amino acid linker, wherein the first amino acid linker is located at the N-terminus of the continuous negatively charged amino acid segments, and the second amino acid linker is located at the C-terminus of the continuous negatively charged amino acid segments, and wherein the first amino acid linker and the second amino acid linker are independently selected from the group consisting of: (a.) Polyglycine linkers (G-linkers) having amino acid sequences (Gly) of length n = 2 to 10. n ; (b.) A glycine-serine connector (GS-connector) comprising at least one glycine and at least one serine, wherein preferably, the GS-connector has (GS) r (G s S) t (GS) u The amino acid sequence, wherein r=0 or 1, s=1-5, t=1-5 and u=0 or 1; and (c.) An amino acid linker (GS*-linker) comprising at least one Gly, at least one Ser and at least one amino acid selected from Thr, Ala, Lys and Cys.

23. The composition for the use of claim 22, wherein the continuous negatively charged amino acid segments consist only of glutamic acid.

24. The composition for the said use as claimed in claim 23, wherein the polypeptide comprising the continuous segments of negatively charged amino acids is composed of SEQ ID NO:49, SEQ ID NO:50 or SEQ ID NO:

51.

25. The composition for the said use according to claims 1 to 24, wherein the CMV VLP comprises the amino acid sequence of SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12, preferably consisting of the amino acid sequence of SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:

12.

26. The composition for the said use according to claims 1 to 25, wherein the NGF antigen is selected from the group consisting of: canine NGF (cNGF), feline NGF (fNGF), equine NGF (eNGF), bovine NGF (bNGF), and suidae NGF (pNGF), wherein preferably the antigen is canine NGF (cNGF) or feline NGF (fNGF), and more preferably, the antigen is canine NGF (cNGF).

27. The composition for the said use according to claims 1 to 26, wherein the NGF antigen comprises or preferably consists of the following: The amino acid sequence is selected from any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33 and SEQ ID NO:55, or has at least 90%, preferably at least 95% sequence identity with any one of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33 and SEQ ID NO:55.

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