MX2007004854A - Gastric inhibitory polypeptide (gip) antigen arrays and uses thereof - Google Patents

Abstract

The present invention is in the fields of medicine, public health, immunology, molecular biology and virology. The present invention provides,inter alia, a composition comprising a virus-like particle (VLP) and at least one antigen, wherein said antigen is a GIP protein or a GIP fragment linked to the VLP respectively. The invention also provides a method for producing the aforesaid composition. The compositions of this invention are useful in the production of vaccines, in particular, for the prevention and/or treatment of obesity and hereby, in particular, by inducing efficient immune responses, in particular antibody responses. Furthermore, the compositions of the invention are particularly useful to efficiently induce self-specific immune responses within the indicated context.

Description

ANTIGEN ARRANGEMENTS OF GASTRIC INHIBITORY POLIPEPTIDE (GIP) AND USES OF THE SAME FIELD OF THE INVENTION The present invention concerns the fields of medicine, public health, immunology, molecular biology and virology. The present invention provides, inter alia, a composition comprising a virus-like particle (VLP) and at least one antigen, where the antigen is a GIP protein (gastric inhibitory peptide, GIP for its acronym in English ) or a GIP fragment linked to the VLP respectively. The invention also provides a method for producing the aforementioned composition. The compositions of this invention are also useful in the production of vaccines, particularly for the prevention and / or treatment against obesity and thereby, particularly by inducing efficient immune responses, particularly antibody responses. Moreover, the compositions of the invention are particularly useful for efficiently inducing self-specific immune responses within the indicated context. BACKGROUND OF THE INVENTION Glucose-dependent insulinotropic polypeptide (GIP, also known as gastric inhibitory peptide) is Ref .: 180038 a gastrointestinal hormone that is released during food from endocrine K cells that line the intestinal wall. The amount of GIP that is released into the bloodstream is largely dependent on the content of the food and is mainly induced by the absorption of ingested amino acids, glucose or fat (Elliott, RM, et al, (1993), J. Endocrinol 138, 159-166, Lardinois, CK et al, (1988), J Am Coll Nutr 7 (3), 241-7.GIP rapidly acts on pancreatic β cells to stimulate insulin vibration, thereby ensuring a rapid insulin-mediated uptake of glucose into tissues (Dupré J. et al, (1973) J. Clin Endocrinol, Metab.37, 826-828) GIP achieves this effect by binding to transmembrane receptor 7 coupled to protein G expressed in ß cells, once bound by GIP, these receptors activate adenylyl cyclase and other transduction signal pathways, ultimately leading to elevated levels of intracellular Ca 2+ and insulin exocytosis (LU, M. et al, (1993), Endocrinology 133, 2861-2870) In addition to the abs Fat and glucose orientation, the ingestion of carbohydrates also stimulates the release of GIP (Elliott, R.M., et al, (1993), J. Endocrinol. 138, 159-166). GIP is considered one of the main incretin factors of the whole-island axis. It has been described that anti-GIP antibodies block the action of GIP on the insulin secretion induced by glucose (Ebert et al, Endocrinology (1982) 111: 1601). Moreover, GIP has also been postulated for acting directly on adipocytes, which express for the GIP receptor (Yip et al, Endocrinology (1998) 139: 4004). Due to its insulinotropic activity, there is considerable interest in using the hormone as a potential therapy for type 2 diabetes (EP171465, WO03 / 030946). Moreover, it has recently been shown that mice with genes deleted for the GIP receptor (GIPR - / -) have higher blood glucose levels with an insufficient initial response to insulin after oral glucose loading. Although blood glucose levels after ingestion did not increase with a high fat diet in GIPR + / + mice due to a greater compensatory insulin secretion, increasing significantly in GIPR - / - mice due to the lack of this potentialization . Consequently, a defect in this enteroinsular axis may contribute to the pathogenesis of diabetes (Miyawaki K, et al, (1999) PNAS 96:26, 14843-14847). The same group of researchers later demonstrated that wild-type mice that eat a high-fat diet show both GIP hypersecretion and an extreme deposit of visceral and subcutaneous fat with insulin resistance. Unlike, mice that lack the receptor GIP (GIPR - / -) that were fed a high-fat diet were protected both from the development of obesity and from insulin resistance (Miyawaki K. et al (2002) Nature Medicine 8: 7, 738-742) . However, several studies have observed decreased insulin secretion and hyperglycemia in rodents after nutrient ingestion when the GIP action is interrupted in an acute manner with antagonists to the GIP receptor (Lewis JT et al, (2000), Endocrinology, 141, 3710-3716; Tseng, CC et al, (1996) J. Clin.Invest.98, 2440-2445). This suggests that chronic treatment with an antagonist for the GIP receptor can result in glucose intolerance or even diabetes (Kieffer, T.J. (2003), Trains in Pharmacological Sciences Vol. 24 No. 3, 110-112). BRIEF DESCRIPTION OF THE INVENTION Surprisingly, we have now discovered that vaccines and inventive compositions, respectively, comprising a GIP protein or a GIP fragment, are capable of inducing marked immune responses, particularly marked antibody responses, leading to high titers. of antibodies against autoantigenic GIP. Moreover, surprisingly we have discovered that inventive compositions and vaccines, respectively, comprising a GIP protein or a GIP fragment are capable of inducing strong immune responses, particularly antibody responses, in obese mice fed a high fat diet both in prophylactic situations and in therapeutic situations. The weight gain of these obese mice that received the vaccines and inventive compositions, respectively, was significantly reduced compared to mice that did not receive the respective compositions and vaccines, respectively. This indicates that the immune responses, particularly the antibodies generated by the inventive compositions and vaccines, respectively, are capable of specifically recognizing GIP in vivo and interfering with its function. Moreover, since the antibodies are large molecules that only inefficiently penetrate the solid tissue, particularly fatty tissue, it was very surprising that the compositions and vaccines of the invention were effective in inhibiting the accumulation of fat storage. Surprisingly, we further discovered that inventive compositions and vaccines, although effective to interfere with GIP function in vivo, particularly to protect recipient animals and prevent them from gaining weight, do not interfere with blood glucose, fructosamine and plasma triglyceride levels, indicating that the compositions of the invention do not lead to diabetes. Therefore, the inventive compositions and vaccines proved to be safe for use in prevention or treatment against Obesity Therefore, in another aspect, the present invention provides a composition comprising (a) a virus-like particle (VLP), with at least one first binding site; and (b) at least one GIP protein or at least one GIP fragment with at least one second binding site, wherein (a) and (b) are linked through the first and second binding sites preferably to form an alignment ordered and repetitive antigens. In preferred embodiments of the invention, virus-like particles suitable for use in the present invention comprise recombinant protein, preferably recombinant coat protein, mutants or fragments thereof or a virus, preferably a bacteriophage RNA. In a preferred embodiment, the inventive composition comprises a GIP fragment. Although a strong immunoprotective response, in particular an antibody response, is assured, the use of GIP fragments for the present invention can reduce a possible induction of self-specific cytotoxic T cell responses and can reduce the cost of production of inventive compositions and vaccines, respectively. In another aspect, the present invention provides a vaccine composition. Moreover, the present invention provides a method for administering the composition of vaccine in a human or animal, preferably in a mammal. The vaccine of the present invention is capable of inducing a strong immune response, particularly an antibody response without the presence of an adjuvant. Therefore, in a preferred embodiment, the vaccine is devoid of any adjuvant. By avoiding the use of an adjuvant, a possible occurrence of undesired inflammatory T cell responses can be reduced. In another aspect, the present invention provides a pharmaceutical composition comprising the inventive composition and a pharmaceutically acceptable carrier. In yet another aspect, the present invention provides a method for effectively preventing, alleviating or treating, particularly obesity. In another aspect, the invention provides a method for the treatment against obesity in an animal, preferably a cat or domestic dog or in a human comprising administering the vaccine of the invention and the VLP-ghrelin vaccine in the same animal or human . In a preferred embodiment of the invention, the vaccine of the invention and the VLP-ghrelin vaccine are administered contemporaneously Jan. Same animal or human. In still another aspect, the invention provides a method for preventing and treating against obesity, preferably preventing obesity in a human comprising administering the vaccine of the invention and the vaccine. of VLP-nicotine in the same human. This is, preferably, to compensate for the increase in body weight after quitting smoking. Also, preferably, this is to compensate for the increase in food intake during and after smoking cessation. The administration of two vaccines in the same animal or human can, additively or preferably synergistically, increase the efficacy of each vaccine when administered individually. BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows the coupling of the GIP fragment in the?) ß VLP in an SDS-PAGE reducing gel. Path M is the molecular marker; Path 1 is the monomer?) ß derivative; Lane 2 is the fragment GIP1-15 GC coupled to the monomer?) ß; Lane 3 is the fragment GIP 31-42 GC coupled to the monomer?) ß. The coupling bands correspond to one, two, three or four peptides coupled per subunit as indicated by the arrows. Figure 2 shows the efficacy of the GIP vaccine 1-15-GC-? ß or vaccine Q ^ -CG-GIP 31-42. C57BL / 6 mice were vaccinated with either murine GIP 1-15-GC, murine CG-GIP-61-42 coupled to?) ß VLP or? ß VLP only as described in detail in Example 6. Figure 2A shows the Weight gain of these mice over a period of time as indicated by the x axis and Figure 2B shows the fat composition of these mice and 142 days after the first immunization. DETAILED DESCRIPTION OF THE INVENTION Antigen: as used herein, the term "antigen" refers to a molecule capable of being bound by an antibody or a T cell receptor (TCR) if it is presented by complex molecules major histocompatibility (MHC). The term "antigen", as used herein, also encompasses T cell epitopes. An antigen is also capable of being recognized by the immune system and / or capable of inducing a humoral immune response and / or a cellular immune response that entails to the activation of B and / or T lymphocytes. However, this may require that, at least in certain cases, the antigen contains or is bound to a Th cell epitope and is provided in the form of an adjuvant. An antigen can have one or more epitopes (B- and T- epitopes). The specific reaction referred to above means that it is indicated that the antigen reacts preferably, usually very selectively, with its corresponding antibody or TCR and not with a multitude of other antibodies or TCRs that may be required by other antigens. The antigens, as used herein, can also be mixtures of several individual antigens.

Antigenic site: The term "antigenic site" and the term "antigenic epitope", which are used here indistinctly, they refer to continuous or discontinuous portions of a polypeptide, which can be linked immunospecifically by an antibody or a T cell receptor within the context of an MHC molecule (main histocompatibility complex). Immunospecific binding excludes non-specific binding but does not necessarily exclude cross-reactivity. The antigenic site typically comprises 5-10 amino acids of a spatial conformation that is unique to the antigenic site. Associated: The term "associated" (or its adjective association) as used herein, refers to all possible forms, preferably chemical interactions, whereby two molecules are joined together. Chemical interactions include covalent and non-covalent interactions. Typical examples of non-covalent interactions are ionic interactions, hydrophobic interactions or hydrogen bonding, while covalent interactions are based, for example, on covalent bonds such as ester, ether, phosphoester, amide, peptide and carbon-phosphorus bonds , carbon-sulfur bonds such as thioether or imide bonds.

Binding site, primer: As used herein, the phrase "first binding site" refers to an element that naturally occurs with the VLP or that is artificially bound to the VLP, and with which the second binding site can be linked .

The first binding site can be a protein, a polypeptide, an amino acid, a peptide, a sugar, a polynucleotide, a natural or synthetic polymer, a dry metabolite or a secondary compound (biotin, fluorescein, retinol, digoxigenin, metal ions and phenylmethylsulfonyl fluoride) ), a group chemically reactive with an amino group, a carboxyl group, a sulfhydryl group, a hydroxyl group, a guanidinyl group, a histidinyl group or a combination thereof. A preferred embodiment of a chemically reactive group that is the first binding site is the amino group of the amino acid lysine. The first binding site is located, usually on the transfer surface of the external surface of the VLP. Multiple first binding sites are present on the surface, preferably on the external surface of the virus-like particle, usually in a repeated configuration. In a preferred embodiment, the first binding site is associated with the VLP, by at least one covalent bond, preferably during at least one peptide bond. Binding site, second: As used herein the phrase "second binding site" refers to an element that naturally exists with or that is artificially added to the GIP of the invention and to which the first binding site can be linked. The second binding site of the GIP of the invention can be a protein, a polypeptide, a peptide, an amino acid, a sugar, a polynucleotide, a natural or synthetic polymer, a compound or secondary metabolite (biotin, fluorescein, retinol, digoxigenin, metal ions and phenylmethylsulfonyl fluoride) or a chemically reactive group such as a amino group, a carboxyl group, a sulfhydryl group, a hydroxyl group, a guanidinyl group, a histidinyl group, or a combination thereof. A preferred embodiment of a chemically reactive group is the second binding site which is the second binding site is the sulfhydryl group, preferably a cysteine amino acid. The terms "GIP protein with at least one second binding site", "GIP fragment with at least one second binding site" or "GIP of the invention with at least one second binding site" refer, therefore, to a structure comprising the GIP of the invention of at least one second binding site. However, in particular for a second binding site, which does not exist naturally with the GIP protein or the GIP fragment, this structure normally and preferably also comprises a "linker". In another preferred embodiment, the second binding site is associated with the GIP of the invention by at least one covalent bond, preferably through at least one peptide bond. Even in another preferred embodiment, the second binding site is artificially added to the GIP of the invention through a amino acid linker, preferably comprising a cysteine by protein fusion. Link: As used herein, the term "link" refers to a link that can be covalent, for example by chemical coupling or coupling or a non-covalent bond, for example ionic interactions, hydrophobic interactions, hydrogen bonds, etc. The covalent bonds can be, for example, ester, ether, phosphoether, amide, peptide, imide bonds, carbon-sulfur bonds, carbon-phosphorus bonds and the like. The term also includes the enclosure or the partial closure of a substance. The term "linked" is broader than and includes terms such as "coupled", "merged", "enclosed", "wrapped", "joined". For example, the polyanionic macromolecule such as polyglutamic acid can be and normally and preferably is enclosed or enveloped in the VLP, normally and preferably without the existence of an actual covalent bond. Cover protein: The term "cover protein" and the term that is used interchangeably "capsid protein" within this application, refers to a viral protein, which is capable of being incorporated into a virus capsid or a VLP. Typically and preferably the term "coat protein" refers to the coat protein encoded for the genome of a virus, preferably a bacteriophage RNA or the genome of a variant of a virus, preferably a bacteriophage RNA. More preferably and by way of example, the term "AP205 coat protein" refers to SEQ ID NO. 14 or the amino acid sequence, wherein the first methionine is cleaved of SEQ ID NO. 14. More preferably and by way of example, the term "? Cover protein" refers to SEQ ID NO: 1 ("? Β CP") and SEQ ID NO: 2 (Al), with or without methion in the term N. The bacteriophage capsid? ß is mainly composed of? ß CP, with a lower content of Al protein. GIP of the invention: The term "GIP of the invention" as used herein, refers to at least one protein GIP or at least one GIP fragment as described herein GIP protein: The term "GIP protein" as used herein should encompass any polypeptide comprising or alternatively or preferably consisting of human GIP of SEQ ID NO: 22, the GIP of mouse of SEQ ID NO: 23, the rat GIP of SEQ ID NO: 24, the bovine GIP of SEQ ID NO: 25, the pig GIP of SEQ ID NO: 26, the cat or dog GIP of SEQ ID NO: 63 or the corresponding GIP sequence of any ortholog from any other animal.The term "ortholog" denotes a polypeptide obtained from of a species that is the functional counterpart of a polypeptide of a different species. The sequence differences between orthologs are the result are the result of speciation. Moreover, the "GIP protein" as used herein, any polypeptide comprising or alternatively or preferably consisting of any naturally occurring or genetically engineered variant having more than 70%, preferably more than 80%, preferably more than 85%, should also encompass , preferably preferably more than 90% and again preferably more than 95% and preferably superlative more than 97% amino acid sequence identity with the human GIP of SEQ ID NO: 22, the mouse GIP of SEQ ID NO: 23, the rat GIP of SEQ ID NO: 24, the bovine GIP of SEQ ID NO: 25, the pig GIP of SEQ ID NO: 26, the cat or dog GIP of SEQ ID NO: 63 or the orthologs corresponding to any other animal. The term "GIP protein" as used herein should also encompass post-translational modifications that include, but are not limited to glycosylations, acetylations, GIP protein phosphorylations as defined above. Preferably, the GIP protein, as defined herein, comprises at least 200 amino acids in length and even more preferably at least 100 amino acids, even more preferably at most 50 amino acids in length. Typically and preferably, the GIP protein linked to the VLP should be able to induce the production of antibodies in vivo specifically capable of binding to the GIPs as can be verified by for example, ELISA.

GIP Fragment: The term "GIP fragment" as used herein should encompass any polypeptide comprising or alternatively or preferably consisting of at least 4, 5, 6, 7, 8, 9, 10, 11 or 12 contiguous amino acids of a protein GIP as defined herein as well as any polypeptide containing more than 65%, preferably more than 80%, preferably more than 85%, more preferably more than 90% even more preferably more than 95% amino acid sequence identity for East. Preferably, the term "GIP fragment" as used herein, should encompass any polypeptide comprising or alternatively or preferably consisting of at least 6 contiguous amino acids of a GIP protein as defined herein as well as any polypeptide having more than 80% , preferably more than 85%, preferably more than 90% and even more preferably more than 95% amino acid sequence identity for it. Preferred embodiments for the GIP fragment are truncation or forms of internal suppression of the GIP protein. Normally and preferably, a GIP fragment linked to the VLP must be able to induce the production of antibodies in vivo, specifically capable of binding with GIP. The amino acid sequence identity of polypeptides can be determined conventionally using computer programs known as the Bestfit program. When it's used Bestfit or any other sequence alignment program, preferably using Bestfit to determine if a particular sequence is, for example, 95% identical to the amino acid reference sequence, the parameters are determined in such a way that the identity percentage is calculated with respect to the entire length of the reference amino acid sequence and which overlaps with respect to homology up to 5% of the total amount of amino acid residues in the reference sequence. This aforementioned method for determining the percent identity between polypeptides is applicable to all proteins, polypeptides or a fragment thereof which are described in the invention. Linked: The term "linked" (or its adjective "link") as used herein, refers to all possible forms, preferably chemical interactions, whereby at least one first binding site and at least one second site of union are joined together. Chemical interactions include covalent and non-covalent interactions. Chemical examples of covalent interactions are ionic interactions, hydrophobic interactions or hydrogen bonding, while covalent interactions are based, for example, on covalent bonds such as ester, ether, phosphoether, amide, peptide bonds, carbon-phosphorus bonds, carbon-sulfur bonds, thioester or imide bonds. In certain preferred embodiments, the first binding site and the second binding site are linked through at least one covalent bond, preferably through at least one non-peptide bond, even more preferably through exclusively non-peptide bonds. doctors The term "linked" as used herein, however, should not only encompass a direct linkage of at least a first binding site and of at least a second binding site but also, alternatively and preferably, an indirect binding of minus a first binding site and at least one second binding site through intermediate molecules and thereby normally and preferably a heterobifunctional crosslinking agent. Linker: A "linker" as used herein, either associates the second binding site with the GIP of the invention or already comprises, essentially consists of, or consists of the second binding site. Preferably, a "linker" as used herein, already comprises the second binding site, normally and preferably, but not necessarily, as an amino acid residue, preferably as a cysteine residue. A "linker" as used herein is also called an "amino acid linker", particularly when the linker according to the invention contains at least one amino acid residue. Therefore, the terms "linker" and "amino acid linker" are used interchangeably herein. However, this does not it implies that this linker comprises exclusively amino acid residues, even if a linker comprising amino acid residues is a preferred way of the present invention. In the amino acid residues of the linker are preferably naturally occurring amino acid compounds or non-natural amino acids known in the art, all L or all D or mixtures thereof. In addition, preferred embodiments of linker according to the invention are molecules comprising a sulfhydryl group or a cysteine residue and therefore, these molecules are also encompassed within the invention. Other linkers useful for the present invention are molecules comprising Ci-C6 alkyl a cycloalkyl such as cyclopentyl or cyclohexyl, a cycloalkenyl, aryl or heteroaryl entity. Moreover, the linkers that preferably comprise a C 1 -C 6 alkyl, C 5 -C 6 cycloalkyl, aryl or heteroaryl entity and additional amino acids can also be used as linkers of the present invention and should be encompassed within the scope thereof. The linker association with GIP of the invention is preferably carried out by at least one covalent bond, more preferably by at least one peptide bond. Orderly and repetitive array or alignment of the antigen: As used herein, the term "ordered and repeating arrangement of the antigen" generally refers to a repeated pattern of the antigen, characterized by normally and preferably a large order of uniformity in a spatial configuration of the antigens with respect to the virus-like particle, respectively. In one embodiment of the invention, the repeated pattern can be a symmetric pattern. Certain embodiments of the invention as VLPs of RNA phage are typical and preferred examples of ordered and repeating arrays of suitable antigens which, furthermore, have orders for strictly repeating antigen crystals, preferably with separations of 1 to 30 nanometers, preferably 2. to 15 nanometers, even more preferably 2 to 10 nanometers, even more preferably 2 to 8 nanometers and more preferably 1.6 to 7 nanometers. Wrapped: The term "wrapped" as used herein, refers to the state of the polyanionic macromolecule in relation to the VLP. The term "enveloped" as used herein includes a bond that may be covalent, which may be, for example, by chemical coupling, or non-covalent, eg, ionic interactions, hydrophobic interactions, hydrogen bonds, etc. The term also includes the enclosure or partial closure of a polyanionic macromolecule. Therefore, the polyanionic macromolecule can be enclosed by the VLP without the existence of a current link, in particular of a covalent bond. In preferred embodiments, at least one polyanionic macromolecule is wrapped within the VLP, more preferably non-covalently. Polypeptide: The term "polypeptide" as used herein, refers to a molecule composed of monomers (amino acids) linearly linked by amide bonds (also known as peptide bonds). This indicates a molecular chain of amino acids and does not refer to a specific length of the product. Therefore, peptides, dipeptides, tripeptides, oligopeptides and proteins are included within the definition of a polypeptide. Post-translational modifications of the polypeptide, for example, glycosylations, acetylations, phosphorylations and the like are also encompassed. Viral particle: The term "viral particle" as used herein, refers to the morphological form of a virus. In some types of viruses they comprise a genome surrounded by a protein capsid; others have additional structures (for example, wrappers, tails, etc.). Virus-like particle (VLP), as used herein, refers to a non-replicative or non-infectious viral particle, preferably non-replicative and non-infective, or refers to a non-replicative or non-infective structure, preferably non-replicative and non-infectious of a viral particle, preferably the capsid of a virus. He The term "non-replicative", as used herein, refers to the fact that it is incapable of replicating the genome comprised by the VLP. The term "non-infectious", as used herein refers to being unable to enter the host cell. Preferably, a virus-like particle according to the invention is non-replicative and / or non-infectious since it lacks all or part of the viral genome or genomic function. In one embodiment, a virus-like particle is a viral particle, wherein the viral genome has been physically or chemically inactivated. Generally and more preferably, the virus-like particle lacks all or part of the replicative and infectious components of the viral genome. A virus-like particle according to the invention may contain nucleic acids other than its genome. A typical and preferred embodiment of a virus-like particle according to the present invention is a viral capsid as the viral capsid of the corresponding virus, bacteriophage, preferably a phage RNA. The terms "viral capsid" or "capsid" refers to the macromolecular structure composed of viral protein subunits. Normally, there are 60, 120, 180, 240, 300, 360 and more than 360 viral protein subunits. In general and preferably, the interactions of these subunits conform to the formation of the viral capsid or the structure similar to a viral capsid in a reorganization inherently repetitive, where the structure is, in general terms, spherical or tubular in shape. RNA-like phage virus-like particle: As used herein, the term "RNA-like phage-like particle" refers to a virus-like particle that comprises or preferably consists essentially of shell proteins, mutants or their fragments of a phage-RNA. In addition, the virus-like particle of an RNA phage resembling the structure of an RNA phage that is non-replicative and / or non-infectious, and that lacks at least the gene or genes encoding the replication machinery of the RNA. phage RNA and that normally also lack the gene or genes that code for the protein or proteins responsible for viral binding or entry into the host. However, this definition should also encompass RNA-like phage virus-like particles, wherein the above-mentioned gene or genes are still present but in inactive form and therefore, they also carry non-replicating virus-like particles and / or do not Infectious of an RNA phage. Within the present description, the term "subunit" and "monomer" are used interchangeably and in equivalent terms within the context. In this application, the term "RNA phage" and the term "bacteriophage RNA" are used interchangeably. One, one or one: When the terms "one", "one" or "one" are used in this description, mean "at least one" or "one or more" unless otherwise indicated. Within this application, antibodies are defined to be specifically enveloped if they bind to the antigen with a binding affinity (Ka) of 106 M "1 or more, preferably 107 M'1 or more, more preferably 108 M" 1 or more. more and more preferably 106 M "1 or more The affinity of an antibody can be readily determined by one skilled in the art (e.g., by a Scatchard analysis.) This invention provides compositions and methods for binding immune responses against GIP in an animal or human The compositions of the invention comprise: (a) a virus-like particle (VLP) with at least one first binding site, (b) at least one antigen with at least one second binding site, wherein at minus the antigen is a GIP protein or a GIP fragment wherein (a) and (b) are linked by at least one first or at least one second binding site, preferably the GIP protein or the GIP fragment is linked to the VLP, in order to form an ordered and repetitive configuration of VLP antigen. In the preferred embodiments of the invention, at least 30, more preferably at least 60, with even more preference at least 120 and more preferably with at least 180 GIP of the invention are linked to VLP. Any virus known in the art to have a Repetitive and ordered structure can be selected as the VLP of the invention. Illustrative RNA or DNA viruses, the capsid or protein of the capsid which can be used for the preparation of the VLPs have been described in WO 2004/009124 page 25, line 10-21, page 26, line 11-28, page 28, line 4 to page 31, line 4. These descriptions are incorporated herein by reference. The virus or virus-like particle can be produced and purified from a cell culture infected with the virus. The virus or virus-like particle resulting for the vaccine purpose needs to be devoid of virulence. A virus or particle similar to avirulent virus can be generated by chemical and / or physical inactivation, such as by UV radiation, treatment with formaldehyde. Alternatively, the genome of the virus can be genetically manipulated by mutation or deletions to render viral replication incompetent. In a preferred embodiment, the VLP is a recombinant VLP. The recombinant VLP as described herein, refers to a VLP that is prepared by a process comprising at least one step of DNA recombination technology. Almost all recognized viruses have been sequenced and are readily available to the public. The gene that codes for the cover protein can be identified easily by the expert. Normally, the envelope protein gene can be cloned by conventional methods into an expression vector and expressed in a suitable host for the vector. VLP is the result of self-assembly of the expressed coat protein and can be further recovered and purified by methods known in the art. Examples have been described in WO02 / 056905 and incorporated herein by reference: host cells suitable for the production of virus-like particles on page 29, line 37 to page 30 line 12; Methods for introducing polynucleotide vectors into host cells on page 30, lines 13-27 and mammalian cells as recombinant host cells for the production of virus-like particles on page 30, lines 28-35. In a preferred embodiment, the virus-like particle comprises or alternatively consists of recombinant proteins, mutants or their fragments, or a virus selected from the group comprising a) phage RNA; b) bacteriophages; c) hepatitis B virus, preferably its capsid protein (Ulrich, et al., Virus Res., 50: 141-182 (1998)) or its surface protein (W092 / 11291); d) measles virus (Warnes, et al., Gene 160: 173-178 (1995)); e) Sindbis virus; f) rotavirus (US 5,071,651 and US 5,374,426; g) foot and mouth disease virus (Twomey, et al., Vaccine 13: 1603-1610, (1995)); h) Norwalk virus (Jiang, X., et al., Science 250: 1580 1583 (1990); Matsui, S.M., et al., J-Clin, Invest. 87: 1456-1461 (1991)); i) alphaviruses; j) retroviruses, preferably their GAG protein (W096 / 30523); k) Ty retrotransposon, preferably the pl protein; 1) human papilloma virus (WO 98/15631); m) polyoma virus; n) tobacco mosaic virus and o) chicken coop virus. In a preferred embodiment, the VLP comprises or consists of more than one amino acid sequence, preferably two amino acid sequences of recombinant proteins, mutants or fragments thereof. The VLP comprises or consists of more than one amino acid sequence and in this application it is referred to as a VLP mosaic. The term "fragment of a recombinant protein" or the term "fragment of a coat protein" as used herein, is defined as a polypeptide, having at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 95% of the length of the wild type recombinant protein or the coat protein, respectively and which preferably retains the ability to form VLP. Preferably, the fragments are obtained by at least one internal deletion, at least one truncation or at least one combination thereof. The term "fragment of a recombinant protein" or "fragment of a coat protein" must also, encompass the polypeptide, which has at least 80%, preferably 90%, or even more preferably 95% amino acid sequence identity with the "recombinant protein fragment" or "fragment of a coat protein", respectively as defined above and that is preferably capable of being assembled into a virus-like particle. The term "recombinant recombinant protein" or the term "mutant of a recombinant protein" as used interchangeably in this invention, or the term "mutant coat protein" or the term "mutant of a coat protein", as used interchangeably in this invention, they refer to a polypeptide having an amino acid sequence derived from the wild-type recombinant protein or the coat protein, respectively, wherein the amino acid sequence is at least 80%, preferably at least 85%, 90 %, 95%, 97% or 99% identical to the wild type sequence and preferably retains the ability to assemble in a VLP. The assembly of the fragment or mutant of the recombinant protein or the coat protein in a VLP can be analyzed, as one skilled in the art will appreciate in expressing the protein in E. coli, optionally purifying the capsids by gel filtration from a cell lysate and analyzing capsid formation in a test of immunodiffusion (Ouchterlony test) or by Electron Microscopy (EM) (Kozlovska, T.M., et al., Gene 137: 133-37 (1993)). The immunodi fusion and EM assays can be carried out directly on a cell lysate. In a preferred embodiment, the virus-like particle of the invention is of hepatitis B virus. The preparation of hepatitis B virus-like particles have been described, inter alia, in WO 00/32227, WO 01/85208 and in WO 01/056905. All three documents are explicitly incorporated herein by way of reference. Other embodiments of HBcAg suitable for use in the practice of the present invention have been described on pages 34-39 of WO 01/056905. In other preferred embodiments of the invention, a lysine residue is introduced into an HBcAg polypeptide to mediate the binding of the GIP of the invention to the VLP of HBcAg. In preferred embodiments, the VLPs and compositions of the invention are prepared using an HBcAg comprising, or alternatively consisting of amino acids 1-44 or 1-149, 1-185 of SEQ ID NO: 20, which are modified so that the amino acids at positions 79 and 80 are replaced with a peptide having the amino acid sequence of Gly-Gly-Lys-Gly-Gly. This modification changes SEQ ID NO: 20 in SEQ ID NO: 21. In other preferred embodiments, the cysteine residues at positions 48 and 110 of SEQ ID NO: 21 or their corresponding fragments, preferably 1-144 or 1-149, are mutated into serine. The invention further includes compositions comprising mutants of the hepatitis B core protein having the corresponding amino acid alterations mentioned above. The invention further includes compositions and vaccines, respectively, comprising HBcAg polypeptides or alternatively consisting of amino acid sequences that are at least 80%, 85%, 90%, 95%, 97% or 99% identical to SEQ ID NO: 21 In another embodiment of the invention, the virus-like particle is a recombinant alphavirus and more specifically a recombinant Sindbis virus. Alphaviruses are positive-strand RNA viruses that replicate their genomic RNA completely in the cytoplasm of the infiltrated cell without a DNA intermediate (Strauss, J. and Strauss, E., Microbiol, Rev. 58: 491-562 (1994)). Several members of the alphavirus family, Sindbis (Schlesinger, S., Biotechnology Trains 11: 18-22 (1993)), Semliki Forest Virus (SFV) (Liljestrom, P. and Garoff, H., Bio / Technology 9: 1356 -1361 (1991)) and others (Davis, NL et al., Virology 171: 189-204 (1989)), have had considerable attention to be used as virus-based expression vectors for a variety of different proteins (Lundstrom, K., Curr Opin Opin Biotechnol 8: 578-582 (1997)) and as candidates for the development of vaccines. In preferred embodiments of the invention, the particle similar to virus of the invention comprises, consists essentially or alternatively consists of recombinant coat proteins, mutants or their fragments and an AR phage. Preferably, the RNA phage is selected from the group comprising a) bacteriophage < 2ß; b) bacteriophage R17; c) bacteriophage fr; d) bacteriophage GA; e) SP bacteriophage; f) bacteriophage MS2; g) Mil bacteriophage; h) bacteriophage MXl; i) bacteriophage NL95; k) bacteriophage f2; 1) bacteriophage PP7 and m) bacteriophage AP205. In a preferred embodiment of the invention, the composition comprises envelope protein, its mutants or RNA phage fragments, wherein the envelope protein has an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 1 and refers to?) ß CP; (b) a mixture of SEQ ID NO: 1 and SEQ ID NO: 2 (referring to the protein ß Al); (c) SEQ ID NO: 3; (d) SEQ ID NO: 4; (e) SEQ ID NO: 5; (f) SEQ ID NO: 6; (g) a mixture of SEQ ID NO: 6 and SEQ ID NO: 7; (h) SEQ ID NO: 8; (i) SEQ ID NO: 9; (j) SEQ ID NO: 10; (k) SEQ ID NO: 11; (1) SEQ ID NO: 12; (m) SEQ ID NO: 13; and (n) SEQ ID NO: 14. Generally speaking, the above-mentioned coat protein is capable of being assembled in a VLP with or without the presence of N-terminal methionine. In a preferred embodiment of the invention, the VLP is a mosaic VLP comprising or alternatively consisting of more than one amino acid sequence, preference two amino acid sequences of coat proteins, their mutants or fragments of a phage RNA. In a preferred embodiment, the VLP comprises or alternatively consists of two different RNA phage coat proteins, these two coat proteins have an amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 2 or of SEQ ID NO: 6 or SEQ ID NO: 7. In preferred embodiments of the present invention, the virus-like particle comprises or alternatively consists essentially of recombinant coat proteins, their mutants or bacteriophage fragments RNA Q3, fr, AP205 or GA. In a preferred embodiment, the VLP of the invention is a VLP of a phage RNA? Β. The capsid of the β-virus-like particle sha capsid structure similar to an eicosahedron phage with a diameter of 25 nm and a quasi-symmetry T = 3. The capsid contains 180 copies of the coated protein that bind in hexamers and covalent pentamers by disulfide bridges (Golmohammadi, R. et al., Structure 4: 543-5554 (1996), which leads to a marked stability of the capsid?) Capsids or VLP that are made from coat proteins? Recombinant ß can, however, contain subunits not linked by disulfide bonds with other subunits within the capsid or be incompletely bound.The capsid or VLP of?) ß shan unusual resistance to organic solvents and denaturing agents. Surprisingly, we have observed that DMSO and acetonitrile concentrations as high as 30% and guanidinium concentrations as high as 1M do not affect the stability of the capsid. The high stability of the capsid or VLP of £ > β is an advantageous feature, particularly for use in the immunization and vaccination of mammals and humans in accordance with the present invention. Other particles similar to preferred phage viruses RNA, in particular of? ß and fr in accordance with this invention are described in WO 02/056905, of which the description is incorporated herein by reference in its entirety. Particularly, example 18 of WO 02/056905 sha detailed description of the preparation of VLP particles of ß. In another preferred embodiment, the VLP of the invention is a VLP of phage AP205 RNA. Competing mutant forms during AP205 VLP assembly, including AP205 cover protein with the substitution of proline at amino acid 5 in threonine, can also be performed in the practice of the invention and lead to other preferred embodiments thereof. WO 2004/007538 describes, in particular in Example 1 and Example 2, how to obtain VLP comprising AP205 cover proteins and thereby particularly the expression and purification thereof. WO 2004/007538 is incorporated herein by reference. AP205 VLPs are highly immunogenic and can be ligated with GIP of the invention to preferentially generate the vaccine structure which shows the GIP of the invention oriented repetitively. The high titre of antibodies is manifested against the GIP shown in the inventions which demonstrates that the linked GIP of the inventions is accessible to interact with antibody molecules and are immunogenic. In a preferred embodiment, the VLP of the invention comprises or consists of a mutant coat protein of a virus, preferably an RNA phage, wherein the mutant coat protein has been modified by removing at least one lysine residue by a substitution and / or by deletion. In another preferred embodiment, the VLP of the invention comprises or consists of a mutant coat protein of a virus, preferably an RNA phage, wherein the mutant coat protein has been modified by the addition of at least one lysine residue by a substitution and / or by an insertion. In a highly preferred embodiment, the mutant coat protein is a phage RNA?) Β, wherein at least or alternatively at least two, lysine residues have been removed by substitution or by deletion. In an alternative mode and very Preferred, the mutant coat protein is a phage RNA? β, wherein at least one or alternatively at least two lysine residues have been added by substitution or by insertion. In another preferred embodiment, the mutant phage coat protein RNA?) Has an amino acid sequence selected from any of SEQ ID NO: 15-19. The deletion, substitution or addition of at least one lysine residue allows the degree of coupling, ie, the amount of GIP of the invention, to be varied by VLP subunits of a virus, preferably phage RNA, in particular to match and customize the vaccine requirements. In a preferred embodiment, the compositions and vaccines of the invention have an antigen density ranging from 0.5 to 4.0. The term "antigen density" as used herein, refers to the average amount of GIP of the invention that is linked per subunit, preferably by envelope protein, of the VLP and more preferably of the VLP of an RNA phage. . Therefore, this value is calculated as the average with respect to all the subunits or monomers of the VLP, preferably of the VLP of an RNA phage in the composition or vaccines of the invention. In another preferred embodiment of the present invention, the virus-like particle comprises or alternatively consists essentially or alternatively consists of a ß mutant coat protein or mutants or fragments thereof and the corresponding Al protein. In another preferred embodiment, the virus-like particle comprises or alternatively consists essentially or alternatively consists of a mutant coat protein with the amino acid sequence SEQ ID NO. 15, 16, 17, 18 or 19 and the corresponding protein Al. Even in another preferred embodiment of the present invention, the virus-like particle comprises or alternatively consists essentially or alternatively consists of a mixture of recombinant coat proteins or fragments thereof or phage RNA?) ß, AP205, fr or GA and recombinant mutant coat proteins or their phage RNA fragments < 2ß, AP205, fr or GA. Competent mutant forms during assembly of AP205 VLPs, including AP205 cover protein with the substitution of proline at amino acid 5 in threonine, asparagine at amino acid 14 in aspartic acid, can also be used in the practice of the invention and they lead to other preferred embodiments thereof: The cloning of AP205Pro-5-Thr and the purification of VLPs are described in WO 2004/007538 and therein, in particular within Example 1 and Example 2. The description of WO document 2004/007538, particularly in Example 1 and Example 2 thereof are explicitly incorporated herein by way of reference. Other RNA phage coat proteins have also been shown to self-assemble upon expression in a bacterial host (Kastelein, RA et al., Gene 23: 245-254 (1983), Kozlovskaya, TM Et al., Dokl. Akad. Nauk SSSR 287: 452-455 (1986), Adhin, MR. Et al., Virology 170: 238-242 (1989), Priano, C. et al., J. Mol. Biol. 249: 283-297 (1995). )). The biochemical and biological properties of GA have been described in particular (Ni, CZ., Et al., Protein Sci. 5: 2485-2493 (1996), Tars, K et al., J. Mol. Biol. 271: 759-773 (1997)) and de fr (Pushko P. et al., Prot. Eng. 6: 883-891 (1993), Liljas, L. et al., J Mol. Biol. 244: 279-290 ( 1994)). The crystal structure of various RNA bacteriophages has been determined (Golmohammadi, R. et al., Structure 4: 543-554 (1996)). By using this information, residues exposed on the surface can be identified and, therefore, the RNA phage coat proteins can be modified so that one or more reactive amino acid residues can be inserted by insertion or substitution. Another advantage of VLP derived from phage RNA is its high production of expression in bacteria that allows the production of a high quantity of material at an economic cost. In a preferred embodiment, the composition of the invention comprises at least one antigen, wherein at least this antigen is a GIP protein or a GIP fragment. In a preferred embodiment, the GIP protein or the GIP fragment is selected from the group comprising: (a) human GIP protein or GIP fragment; (b) Bovine GIP protein or GIP fragment; (c) sheep GIP protein or GIP fragment; (d) dog GIP protein or GIP fragment; (e) feline GIP protein or GIP fragment; (f) mouse GIP protein or GIP fragment; (g) Pig GIP protein or GIP fragment; (h) chicken GIP protein or GIP fragment; (i) horse GIP protein or GIP fragment and (g) rat GIP protein or GIP fragment. In a preferred embodiment, at least one antigen is a GIP protein. In another preferred embodiment, the GIP protein comprises, consists essentially of or consists of an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 22; (b) SEQ ID NO: 23; (c) SEQ ID NO: 24; (d) SEQ ID NO: 25; (e) SEQ ID NO: 26; (f) SEQ ID NO: 63; (g) the orthologs corresponding to the GIP from any other animal; and (h) an amino acid sequence with at least 80% or preferably at least 85%, more preferably with at least 90%, with at least 95% identical preference with any of SEQ ID NO: 22-26 or with SEQ ID NO: 63 In a preferred embodiment, the GIP protein comprises, consists essentially of or consists of an amino acid sequence wherein at most 7, preferably 6, 5, 4, more preferably at most 3, 2 or 1 amino acids are deleted, insert or replace, preferably through conservative substitutions compared to the amino acid sequence selected from the group of: (a) SEQ ID NO: 22; (b) SEQ ID NO: 23; (c) SEQ ID NO: 24; (d) SEQ ID NO: 25; and (e) SEQ ID NO: 26. In a preferred embodiment, at least one antigen is a GIP fragment, wherein the GIP fragment comprises or alternatively consists of at least one antigenic site. Methods for determining the antigenic sites of a protein are known to the person skilled in the art. Document PCT / EP2005 / 004980 has developed some of these methods from the first paragraph on page 26 through the fourth paragraph on page 27 and these specific descriptions are incorporated herein by reference. It should be noted that these methods are generally applicable to other polypeptide antigens and therefore, are not restricted to IL-23 pl9 as described in PCT / EP2005 / 004980. In a preferred embodiment of the present invention, the GIP fragment comprises or alternatively or preferably consists of at least 5 to 12 contiguous amino acids of a GIP protein as defined herein. In another preferred embodiment, the GIP fragment is selected from the amino part of GIP. The amino part of GIP, as used herein, refers to the first 18, preferably the first 15 amino acid sequences of GIP of SEQ ID NO: 22 or the corresponding orthologs of any other animal. In a preferred embodiment, the GIP fragment comprises, consists essentially of or consists of the 7-10 amino acid sequence (SEQ ID NO: 64), preferably 4-10 (SEQ ID NO: 67), preferably 4-13 (SEQ ID NO. : 32), preferably 1-10 (SEQ ID NO: 29), preferably 4-11 (SEQ ID NO: 45), preferably 7-15 (SEQ ID NO: 65), more preferably 4-15 (SEQ ID NO 66) ), even more preferably 1-15 (SEQ ID NO: 27), of SEQ ID NO: 22 or the corresponding GIP orthologs of any other animal. In a preferred embodiment, the GIP fragment comprises, consists essentially of or consists of the 7-10 amino acid sequence (SEQ ID NO: 64), preferably 4-10 (SEQ ID NO: 67), preferably 4-13 (SEQ ID NO. : 32), preferably 1-10 (SEQ ID NO: 29), preferably 4-11 (SEQ ID NO: 45), preferably 7-15 (SEQ ID NO: 65), more preferably 4-15 (SEQ ID NO 66) ), even more preferably 1-15 (SEQ ID NO: 27), of SEQ ID NO: 22 or the corresponding GIP orthologs of any other animal, wherein an amino acid of sequence 7-10 or where 3, preferably 2 , even more preferably 1 amino acid of 4-10, 4-13, 1-10, 4-11, 4-15, 7-15 and 1-15, amino acid sequences, have been or will be mutated, preferably by deletion, insertion and / or substitution, with greater preference through conservative substitutions. Conservative substitutions, as he understands those skilled in the art include isosteric substitutions, are substitutions where the charged, polar, aromatic, aliphatic or hydrophobic nature of the amino acid is maintained. Typical conservative substitutions are substitutions between amino acids within one of the following groups: Gly, Ala; Val, lie, Leu; Asp, Glu; Asn, Gln; Ser, Thr, Cys; Lys, Arg; and Phe and Tyr. In a preferred embodiment, the GIP fragment comprises, consists essentially of or consists of an amino acid sequence selected from the group comprising (a) SEQ ID NO: 27; (b) SEQ ID NO: 29; (c) SEQ ID NO: 32; (d) SEQ ID NO: 45 and (e) an amino acid sequence with at least 80% or preferably at least 85%, more preferably at least 90%, more preferably at least 95% identity with SEQ ID NO: 27 , 29, 32 or 45. In a preferred embodiment, the GIP fragment comprises, consists essentially of or consists of amino acids 20-23, more preferably 19-25, more preferably 16-30 of SEQ ID NO: 22-26 or of SEQ ID NO: 63 of the GIP sequence of the corresponding orthologs of any animal. In another preferred embodiment, the GIP fragment comprises, consists essentially of or consists of an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 31; (b) SEQ ID NO: 43 and (e) an amino acid sequence with at least 80% or preferably at least 85%, plus preferably at least 90% or more preferably at least 95% identity with SEQ ID NO: 31 or 43. In a further preferred embodiment, the GIP fragment comprises, consists essentially of or consists of amino acid 20-23, preferably 19- 25, more preferably 16-30 of SEQ ID NO: 22-26 or SEQ ID NO: 63 of the GIP sequence of the corresponding orthologs of any other animal, wherein an amino acid of sequence 20-23 or wherein , preferably 2 or even more preferably an amino acid of 19-25, preferably 16-30 imitated amino acid sequences, preferably by deletion, insertion and / or substitution, more preferably by conservative substitutions. In a preferred embodiment, the GIP fragment comprises, consists essentially or consists of amino acid 31-34, preferably 30-34, more preferably 28-34, more preferably 30-37, more preferably 28-37, more preferably 31-42 , more preferably 28-42 of SEQ ID NO: 22-26 or SEQ ID NO: 63 or of the GIP sequence of the corresponding orthologs of any other animal. In another preferred embodiment, the GIP fragment comprises, consists essentially of or consists of an amino acid sequence selected from the group consisting of: (a) SEQ ID NO: 28; (b) SEQ ID NO: 44; (c) SEQ ID NO: 68; and (d) an amino acid sequence having at least 80%, preferably at least n85%, more preferably at least 90% or more preferably at least 95% identity with SEQ ID NO: 28, 44 or 68. In a preferred embodiment, the GIP fragment comprises or consists essentially of amino acid 31-34, preferably 30-34, more preferably 28-34, more preferably 26-34, more preferably 30-37, more preferably 28-37, more preferably 26-37, even more preferably 31-42, more preferably 28-42, more preferably 26-42 of SEQ ID NO: 22-26 or SEQ ID NO: 63 of the GIP sequence of the corresponding orthologs of any other animal, wherein an amino acid of the sequence 31-34 or 30-34 or where 3, preferably 2 and even more preferably 1 amino acid of the sequence 28-34, 30-37, 28-37, 31-42 or 28-42 has been or will be mutated, preferably by deletion, insertion and / or substitution, more preferably by conservative substitutions. In a preferred embodiment, the GIP fragment or the GIP protein further comprises a stretch of hydrophilic amino acids fused to the GIP fragment or the GIP protein as described above. A stretch of hydrophilic amino acids as used herein, refers to a stretch of amino acids with at least 60%, preferably at least 70%, more preferably at least 80% or even more preferably at least 90% amino acids being hydrophilic amino acids . In a preferred embodiment, the stretch of amino acids consists of at least 7, preferably 6, 5, 4, more preferably 3, more preferably 2 or one amino acid. The addition of hydrophilic amino acids increases the solubility of the GIP fragment or the GIP protein. In another preferred embodiment, the hydrophilic amino acids are glycine, serine, threonine and the charged amino acids. In another preferred embodiment, the charged amino acids are lysine, arginine, aspartic acid, glutamic acid. In a preferred embodiment, the amino acid stretch comprises or consists of the amino acid sequence DD, KK, RR, DE, ED, EE, KR, RK. In a preferred embodiment, the GIP fragment comprises or consists essentially of the amino acid sequence selected from the group comprising: (a) amino acids 1-12 of SEQ ID NO: 22 with DD or KK added at the C-terminus; (b) amino acids 1-13 of SEQ ID NO: 22 with DD or KK added at the C-terminus; (c) amino acids 1-14 of SEQ ID NO: 22 with DD or KK added at the C-term (SEQ ID No. 69 and 70); (d) amino acids 1-15 of SEQ ID NO: 22 with DD or KK added in the C-terminus (SEQ ID No. 71 and 72) and (e) amino acids 1-11 of SEQ ID NO: 22 with DD added or KK in the C-term. In one embodiment, at least one antigen of the invention comprises at least two GIP fragments, preferably two GIP fragments. In a preferred embodiment, two GIP fragments are two different GIP fragments. In a modality preferred, the two GIP fragments are fused into a single polypeptide. In one embodiment, the two GIP fragments are directly fused. In another embodiment, the two GIP fragments are fused through a separator sequence. In a preferred embodiment of the invention, the VLP with at least one first binding site is linked to the GIP of the invention with at least one second binding site via at least one peptide bond. The gene encoding the GIP of the invention, preferably a GIP fragment, more preferably a fragment not greater than 50 amino acids, even more preferably less than 30 amino acids, is ligated into the framework, either internally or preferably with the N- or the C-terminus to the gene that codes for the VLP cover protein. The embodiments of fusing antigens of the invention in the coat protein, mutants or fragments thereof in a virus, preferably an RNA phage, have been described in WO 2004/009124 page 62 line 20 to page 68 line 17 and are incorporated here for reference. The fusion protein should preferably retain the ability to assemble into a VLP at the time of expression which can be examined by electron microscopy. Flanking amino acid residues can be added to increase the distance between the coat protein and the foreign epitope. The glycine and serine residues are particularly favorable amino acids for used in the flanking sequences. This flanking sequence confers additional flexibility that may decrease the potential destabilizing effect of fusing a foreign sequence in the sequence of a VLP and subunit. decrease the interference with the assembly by the presence of the foreign epitope. In another preferred embodiment, the GIP of the invention, preferably GIP fragments, even more preferably GIP fragments with amino acids of sequence SEQ ID NO: 27-32, SEQ ID NO: 43-45, SEQ ID NO: 66 or 68 are fused either to the No C-terminus of a coat protein, its mutants or fragments of an AP205 phage RNA. In another preferred embodiment, the fusion protein further comprises a spacer, wherein the spacer is located between the coat protein, its fragments or mutants of the AP205 and the GIP of the invention. In a preferred embodiment of the present invention, the composition comprises or alternatively consists essentially of a virus-like particle with at least one first binding site ligated with at least one GIP of the invention with at least one second binding site by means of at least one covalent bond, preferably the covalent bond is a non-peptide bond. In a preferred embodiment of the present invention, the first binding site comprises or is preferably, an amino group, preferably the amino group of a lysine residue. In Another preferred embodiment of the present invention, the second binding site comprises or preferably is, a sulfhydryl group, preferably a sulfhydryl group of cysteine. In another preferred embodiment of the present invention, the second binding site comprises or preferably is a maleimido group that is associated, preferably, covalently associated with at least one antigen. In a highly preferred embodiment of the invention, at least one first binding site comprises or is preferably an amino group, preferably an amino group of a lysine residue and at least the second binding site comprises or is preferably a sulfhydryl group, preferably a Sulfhydryl group of cysteine. In a preferred embodiment of the invention, the GIP of the invention is linked to VLP by chemical crosslinking, normally and preferably by the use of a heterobifunctional crosslinking agent. In preferred embodiments, the heterobifunctional crosslinking agent contains a functional group that can react with the first preferred binding sites, preferably with the amino group, more preferably with the amino groups of VLP lysine residues and another functional group that can react with the second preferred binding site, i.e., a sulfhydryl group, preferably of cysteine residues inherent thereto or that are artificially added to the site of the invention or optionally also made available to react by reduction. Several heterobifunctional crosslinking agents are known in the art. These include the preferred crosslinking agents SMPH (Pierce), Sulfo-MBS, Sulfo-EMCS, Sulfo-GMBS, Sulfo-SIAB, Sulfo-SMPE-, Sulfo-SMCC, SVSB, SIA and other crosslinking agents available for example, from Pierce Chemical Company and having a functional group reactive towards amino groups and an amino functional group reactive towards sulfhydryl groups. The aforementioned crosslinking agents all lead to the formation of an amide bond after the reaction with the amino group and a thioether bond with the sulfhydryl groups. Another class of crosslinking agents suitable for the practice of the invention are characterized by the introduction of a disulfide bond between the GIP of the invention and the VLP at the time of coupling. Preferred crosslinking agents belonging to this class include, for example, SPDP and Sulfo-LC-SPDP (Pierce). In a preferred embodiment, the composition of the invention further comprises a linker. The genetic manipulation of a second binding site in the GIP of the invention is achieved by the association of a linker, preferably containing at least one suitable amino acid as a second binding site, in accordance with the descriptions of this invention, therefore, in a preferred embodiment of the present invention, a linker is associated with the GIP of the invention by at least one covalent bond, preferably by at least one a peptide bond. Preferably, the linker comprises or alternatively consists of a second binding site. In another preferred embodiment, the linker comprises a sulfhydryl group, preferably a cysteine residue. In another preferred embodiment, the amino acid linker is a cysteine residue. The selection of a linker will depend on the nature of the GIP of the invention or its biochemical properties such as pl, charge distribution and glycosylation. In general terms, flexible amino acid linkers are favored. In another preferred embodiment of the present invention, the linker comprises amino acids, wherein also preferably, the linker comprises as much as possible, preferably as much as 20, more preferably as much as 15 amino acids. Preferred embodiments of the linker are selected from the group comprising: (a) CGG or CG / GC; (b) 1 N-terminal gamma linker (e.g., CGDKTHTSPP, SEQ ID NO: 48); (c) N-terminal gamma 3 linker (e.g., CGGPKPSTPPGSSGGAP, SEQ ID NO: 59); (d) hinge Ig regions; (e) N-terminal glycine linkers (e.g.

GCGGGG, SEQ ID NO: 49), (f) (G) kC (G) n (with n = 0-12 and k = 0-5); (g) N-terminal glycine-serine linkers ((GGGGS) n, n = 1-3 with another cysteine plus (eg, SEQ ID NO: 50, corresponding to a modality where n = 1); (h) ( G) kC (G) m (S) 1 (GGGGS) n with n = 0-3, k = 0-5, m = 0-10, 1 = 0-2 (for example, SEQ ID NO. corresponds to a modality where n = 1, k = 1, 1 = 1 and m = 1); (i) GGC (k) GGC-NH2; (1) C-terminal gamma 1 linker (eg, DKTHTSPPCG, SEQ ID NO : 52); (m) C-terminal gamma 3 linker (eg, PKPSTPPGSSGGAPGGCG, SEQ ID NO: 53); (n) C-terminal glycine linkers (GGGGCG, SEQ ID NO: 54); (o) (G) nC (G) k with n = 0-12 and k = 0-5; (p) C-terminal glycine-serine linkers ((SGGGG) nn = 1-3 with another additional cysteine (eg, SEQ ID NO: 55, which corresponds to a modality where n = 1); (q) (G) m (S) 1 (GGGGS) n (G) oC (G) k with n = 0-3, k = 0-5, m = 0 -10, 1 = 0-2 yo = 0-8 (for example, SEQ ID NO: 56, which corresponds to a modality where n = 1, k = l, 1 = 1, o = 1 and m = 1). another preferred embodiment, the linker is added to the N-terminus of the GIP of the invention. In another preferred embodiment of the invention, the linker is added to the C-terminus of the GIP of the invention. Preferred linkers according to the invention are glycine (G) n linkers which also contain a cysteine residue as a second binding site, such as the N-terminal glycine linker (GCGGGG) and the C-terminal glycine linker (GGGGCG) Other preferred embodiments are the C-terminal glycine-lysine linker (GGKKGC; SEQ ID NO: 57) and the N-terminal glycine-lysine linker (CGKKGG; SEQ ID NO: 58), GGCG to GGC or GGC-NH2 ("NH2"). "means amidation) and are linkers in the C-terminus of the peptide or CGG in the N-terminus. In general terms, the glycine residues are inserted between heavy amino acids and the cysteine is used as the second binding site to avoid a steric hindrance of the heaviest amino acid in the coupling reaction. In a preferred embodiment, the sequence of the linker is GC, preferably GC is associated with the C-terminus of the GIP protein or the GIP fragment, preferably with SEQ ID NO: 22-27k, 29, 31-32, 43 or 45. In another preferred embodiment, the sequence of the linker is CG, preferably CG is fused to the N-terminus of the GIP protein or the GIP fragment, preferably to SEQ ID NO: 28, 44 or 68. The cysteine residues serve as the second binding site, having to be available either inherently or in addition to the GIP of the invention must be in the reduced state to react with the heterobifunctional crosslinking agent in the activated carrier which is a free cysteine or a cysteine residue with a free sulfhydryl group. The binding of the GIP of the invention to the VLP by the use of a heterobifunctional crosslinking agent according to the preferred methods described above, allows the coupling of the GIP of the invention with the VLP in oriented form. Other methods for joining the GIP of the invention to the VLP include methods where the GIP of the invention is cross-linked with the VLP, using carbodiimide EDC and NHS. The GIP of the invention can also be violated by reaction, for example with SATA, SATP or iminothiolane. The GIP of the invention, after being checked out, if required, can be coupled with the VLP in the following manner. After removing excess rape reagent, the GIP of the invention is reacted with the VLP, previously activated with a heterobifunctional crosslinking agent comprising a reactive cysteine entity and therefore showing at least one or several reactive functional groups towards cysteine residues with which the thiolated GIP of the invention may react as described above. Optionally, small amounts of reducing agent are included in the reaction mixture. In other methods, the GIP of the invention binds to VLP, using a homobifunctional crosslinking agent such as glutaraldehyde, DSG, BM [PE0] 4, BS3, (Pierce) or other known homobifunctional crosslinking agents with functional groups reactive toward groups amine or carboxyl groups of the VLP. In a preferred embodiment, the first binding site comprises or is preferably a sulfhydryl group, even more preferably a sulfhydryl group of a cysteine which is added naturally or artificially to the cover protein or alternatively consisting essentially of the VLP. The second binding site is the maleimido group of a linker, such as MBS (m-maleimidobenzoyl-N-hydroxysuccinimide ester) or SMPH (succinimidyl-6- [β-maleimidopropionamido] hexanoate), which is chemically associated with the antigen, preferably covalently associated with the antigen, more preferably covalently associated with an amino group of the antigen, even more preferably covalently associated by an NHS-ester group of the linker with the amino group of the amino acid N-terminus of the antigen. In a preferred embodiment, at least one antigen, preferably no more than 50, even more preferably only 30 amino acids, even more preferably GIP protein or GIP fragment of SEQ ID NO: 22-27, 29, 31-32, 43 or 45 are chemically synthesized and the maleimido group is preferably associated with the amino group of the amino acid N-terminus. The first binding site and the second binding site are linked by a thioether bond. In a preferred and alternative embodiment, the first binding site comprises or is preferably an amino group, preferably an amino group of natural lysine or artificially added to the covering protein comprising or alternatively consisting essentially of VLP. The second binding site is the maleimido group of a linker as elaborated in the previous paragraph. The amino group of the VLP is derivatized by a heterobifunctional crosslinking agent, such as N-succinimidyl-S-acetylthioacetate (SATA) or 2-iminothiolane, in a sulfhydryl group, which is then reactive to the maleimido group of the linker. Preferred linkers comprising at least one maleimido group are, for example, SMPH, Sulfo-MBS. Other preferred linkers are Sulfo-EMCS, Sulfo-GMBS, Sulfo-SIAB, Sulfo-SMPB, Sulfo-SMCC, SVSB, SIA and other cross-linking agents available, for example, from Pierce Chemical Company and having a reactive functional group to amino groups and a functional group reactive towards sulfhydryl groups. In other embodiments of the present invention, the composition comprises or alternatively consists essentially of a virus-like particle bound to the GIP of the invention by means of chemical interactions, wherein at least one of these interactions is not a covalent bond. For example, the binding of the VLP to the GIP of the invention can be effected by biotinylation of the VLP and expression of the GIP of the invention as a fusion protein-streptavidin. Other binding pairs, such as ligand-receptor, antigen-antibody can also be used as a coupling reagent in a similar manner as biotin-avidin.

US 5,698,424 discloses a modified bacteriophage coat protein MS-2 capable of forming a capsid, wherein the coat protein is modified by the insertion of a cysteine residue in the N-terminal hairpin region by the replacement of each of the cysteine residues located externally to the N-terminal hairpin region by an amino acid residue that is not cysteine. The inserted cysteine can be ligated directly into the desired molecular species to be presented as an epitope or an antigenic protein. However, we observed that the presence of a cysteine-free residue exposed in the capsid leads to the oligomerization of capsids in the form of disulfide bridge formation. Moreover, the union between capsids and antigenic proteins by disulfide bonds is labile, in particular the molecules that contain sulfhydryl entities and, moreover, are less stable in the soil than, for example, the thioether bonds (Martin FJ and Papahadjopoulos D (1982). Irreversible Coupling of Immunoglobulin Fragments to Preformed Vesicles, J. Biol. Chem. 257: 286-288). Therefore, in another highly preferred embodiment, the binding of the VLP and at least one antigen does not comprise a disulfide bond. Also preferred herein is at least one second linkage comprising or preferably being a sulfhydryl group. Moreover, in another very preferred embodiment of the invention, the binding of the VLP and at least one antigen does not comprise a sulfur-sulfur bond. In another highly preferred embodiment, at least one first binding site is not or does not comprise a sulfhydryl group of cysteine. Again, in another highly preferable embodiment, at least one first binding site is not or does not comprise a sulfhydryl group. In a highly preferred embodiment of the invention, VLP is produced recombinantly in a host and wherein the VLP is essentially free of host RNA, preferably nucleic acids in the host or where the VLP is essentially free of host DNA, preferably nucleic acids of the host. In a preferred embodiment, the VLP of an RNA phage is produced recombinantly in a host and where the VLP of an RNA phage is essentially free of host RNA, preferably host nucleic acids. In another preferred embodiment, the composition further comprises at least one polyanionic macromolecule linked or preferably enveloped or enclosed in the VLP. In yet another preferred embodiment, the polyanionic macromolecule is polyglutamic acid and / or polyaspartic acid. In a preferred embodiment, the VLP is an RNA phage. The reduction or elimination of the amount of host RNA, preferably nucleic acids from the host, minimizes or reduces unwanted responses of T cells in response to T cells. inflammatory and cytotoxic T cell responses and other unwanted side effects such as fever, while at the same time maintaining a high antibody response, specifically against GIP. Essentially free of host RNA (or host DNA), preferably host nucleic acids: the term "essentially free of host RNA (or host DNA), preferably host nucleic acids" as used herein, refers to the amount of host RNA (or host DNA), preferably host nucleic acids, comprised in the VLP, which normally and preferably is less than 30 g, preferably less than 20 μg, more preferably less than 10 μg, even more preferably less than 8 μg, even more preferably less than 6 g, even more preferably less than 4 g, and more preferably less than 2 μg per mg of VLP. The term host, as used within the aforementioned context, refers to the host where the VLP is produced recombinantly. The person skilled in the art knows the conventional methods for determining the amount of RNA (or DNA), preferably nucleic acids. The typical and preferred method for determining the amount of RNA, preferably nucleic acids, according to the present invention is described in Example 17 of PCT / EP2005 / 055009 filed on October 5, 2005 by the same assignee. Typically and analogously, identical, similar or analogous conditions are used for determining the amount of RNA (or DNA), preferably nucleic acids for inventive compositions comprising VLPs other than?) ß. The modifications of the possibly necessary conditions are within the knowledge of the person skilled in the art.

The term "polyanionic macromolecule" as used herein refers to a molecule of high relative molecular mass which comprises repetitive groups of negative charge, structure essentially comprises multiple repetitions of units derived, actually or conceptually, from molecules of low molecular mass relative. In one aspect, the invention provides a vaccine composition comprising the composition of the invention. In a preferred embodiment, the GIP of the invention linked to the VLP in the vaccine composition is of animal origin, preferably mammalian or human. In other preferred embodiments, the GIP of the invention is of human, bovine, chicken, dog, cat, mouse, rat, pig or horse origin.

In a preferred embodiment, the vaccine composition further comprises at least one adjuvant. The administration of at least one adjuvant can occur before, concomitantly or subsequent to the administration of the inventive composition. The term "adjuvant" as used herein, refers to non-specific stimulators of the immune response or substances that allow generation of a depot in the host which when combined with the vaccine and pharmaceutical composition, respectively, of the present invention, it can even provide a more potent immune response. A variety of adjuvants can be used. Examples include complete and incomplete Freund's adjuvant, aluminum hydroxide and modified muramyldipeptide. Other adjuvants are mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin (limpet hemocyanin), dinitrophenol and potentially useful human adjuvants such as BCG (bacille Calmette Guerin) and Corynebacterium parvum These adjuvants are also well known in the art. Other adjuvants that can be administered with the compositions of the invention include, among others, monophosphoryl lipid immunoregulator Adju Vax 100a, QS-21, QS-18, CRL1005, Aluminum salts (Alum), MF-59, OM-174, OM -197, OM-294 and Virosomal adjuvant technology. The adjuvants may also comprise a mixture of these substances. In another preferred embodiment, the vaccine composition is devoid of adjuvant. An advantageous feature of the present invention is the high immunogenicity of the composition, even in the absence of adjuvants. The absence of an adjuvant, moreover, minimizes the occurrence of undesired responses of inflammatory T cells that represent a concern about safety in vaccination against autoantigens. Therefore, administration of the vaccine of the invention in a patient preferably occurs without the administration of at least one adjuvant in the same patient prior to, concomitantly with or subsequent to the administration of the vaccine. In another aspect, the present invention provides the use of a composition comprising (a) a virus-like particle with at least a first binding site and (b) at least one non-human GIP, preferably a non-human vertebrate of the invention with at least one second binding site, wherein (a) and (b) are joined by at least a first site and a second site for the manufacture of a medicament for the treatment against diseases related to GIP, particularly against obesity in humans. These preferred embodiments comprising at least a non-human GIP of the invention, for example, GIP feline, canine, bovine, rat or mouse of the invention are capable of inducing antibody responses cross recognizing GIP human. The invention further discloses an immunization method which comprises administering the vaccine of the present invention to an animal or a human. Preferably the animal is a mammal such as a cat, sheep, pig, horse, bovine, dog, rat, mouse and particularly a dog or a cat, preferably a domestic cat. The vaccine can be administered in an animal or human by various methods known in the art, but is usually administered by injection, infusion, inhalation, oral administration or other suitable physical methods. The conjugates can alternatively be administered intramuscularly, intravenously, transmucosally, transdermally, intranasally, intraperitoneally or subcutaneously. The conjugate components for administration include suspensions and sterile aqueous solutions (eg, physiological saline) or non-aqueous solutions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil and injectable organic esters such as ethyl oleate. Occlusive carriers or dressings can be used to increase skin permeability and enhance antigen absorption. It is said that the vaccines of the invention are "pharmacologically acceptable" and their administration can be tolerated by a recipient patient. In addition, vaccines of the invention are administered in a "therapeutically effective amount" (ie, an amount that produces the effect physiological desired). The nature or type of immune response is not a limiting factor in its description. Without intending to limit the present invention by the following mechanistic explanation, the inventive vaccine could induce antibodies that bind with the GIP and thereby reduce its concentration and / or interfere with its pathological or physiological function. In another aspect, the invention provides a pharmaceutical composition comprising the composition as shown in the present invention and a pharmaceutically acceptable carrier. When the vaccine of the invention is administered in a patient, it may be in a form that contains salts, buffers, adjuvants or other substances that are desirable to improve the effectiveness of the conjugate. Examples of materials suitable for use in the preparation of pharmaceutical compositions are provided in various sources including REMINGTON'S PHARMACEUTICAL SCIENCES (Osol, A, ed., Mack Publishing Co., (1990)). In another aspect, the invention provides a method for producing the composition of the invention or the vaccine composition of the invention or the pharmaceutical composition of the invention, wherein the method comprises: (a) providing a VLP with at least one first site of Union; (b) providing at least one antigen, wherein the antigen is a GIP protein or a GIP fragment with at least a second site of Union; and (c) linking the VLP to at least one antigen by at least one first binding site and at least one second binding site to produce the composition. In another preferred embodiment, the step of providing a VLP with at least one first binding site further comprises the steps of: (a) disassembling the virus-like particle in the coat proteins, their mutants or their defined fragments, preferably a bacteriophage RNA; (b) purifying these coat proteins, their mutants or fragments; (c) reassembling the purified coat proteins, their mutants or fragments of the virus, preferably a bacteriophage RNA in a virus-like particle, wherein the virus-like particle is essentially free of host RNA (or DNA), preferably nucleic acids of the host. In still another preferred embodiment, the reassembly of the purified coating proteins is carried out in the presence of at least one polyanionic macromolecule, preferably polyglutamic acid and / or polyaspartic acid. Again in one aspect, the invention provides compositions that can be used to prevent, treat and / or attenuate diseases or conditions wherein the GIP exerts an important pathological function, in particular obesity. In another aspect, the invention provides a kit comprising at least one first composition and at least one second composition, wherein the first composition comprises: (a) a first virus-like particle (VLP) with at least one first binding site; (b) at least one first antigen with at least one second binding site, wherein at least one first antigen is a GIP protein or a GIP fragment and wherein (a) and (b) are linked through at least one first and second junction sites; and wherein the second composition comprises: (c) a second virus-like particle (VLP) with at least one first binding site; (d) at least a second antigen with at least one second binding site, wherein at least one second antigen comprises or is a molecule selected from the group comprising: (i) a ghrelin or a ghrelin peptide; (ii) a nicotine, a cotinine or a nornicotine; and (iii) a second GIP protein or a second GIP fragment, wherein the second GIP protein or GIP fragment is different from the first GIP protein or first GIP fragment comprising the first composition and wherein (c) and (d) are link through at least one first and second binding sites. In preferred embodiments, the first composition is the composition of the invention, the first virus-like particle is the virus-like particle of the invention and the first antigen is the GIP of the invention. Also, the second virus-like particle is the virus-like particle of the invention. Therefore, the modalities Preferred with this feature and preferred embodiments, the preparation and use of the first composition, the first virus-like particle and the first antigen have been described and defined in this invention. In a preferred embodiment, the first VLP and the second VLP are different. In a preferred embodiment, the first VLP and the second VLP are equal. In a preferred embodiment of the invention, the first and / or the second virus-like particle is / are an RNA phage, wherein, preferably, the phage RNA is a phage RNA?) ß, fr, GA or AP205. In a preferred embodiment, the virus-like particle and / or the second virus-like particle comprise or consist of recombinant proteins, mutants or fragments thereof of an RNA phage, wherein, preferably the phage RNA is an RNA phage. 2ß, fr, GA or AP205. in another preferred embodiment, the first and second VLPs are both an RNA phage, preferably a phage RNA?) ß. In a preferred embodiment, the first and second VLPs of a virus, preferably an RNA phage, are / are recombinantly produced in a host where the first or the second VLP is essentially free of host RNA, preferably free of the host nucleic acid. In a preferred embodiment, the ghrelin or ghrelin peptide is selected from the group comprising: (a) ghrelin peptide or human ghrelin; (b) ghrelin peptide or dog ghrelin; (c) ghrelin peptide or cat ghrelin; (d) ghrelin peptide or bovine ghrelin; (e) ghrelin peptide or pig ghrelin; (f) peptide ghrelin or sheep ghrelin; and (g) ghrelin peptide or mouse ghrelin. In another preferred embodiment, the ghrelin or ghrelin peptide consists essentially or consists of an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 33; (b) SEQ ID NO: 354; (c) SEQ ID NO: 40; (d) SEQ ID NO: 36; (e) SEQ ID NO: 37; (f) SEQ ID NO: 38; (g9 SEQ ID NO: 46 (GSSFLSPEHQKLQ); and (h) SEQ ID NO: 47 (GSSFLSPEHQKVQ) The preparation and use of a preferred embodiment of the second composition, wherein at least a second antigen is ghrelin or ghrelin peptide, have been described in the patent application WO 2004/009124 and in WO2005 / 068639, both complete descriptions are incorporated herein by reference.Other preferred embodiments of the second composition of the present invention are explicitly defined in the claims of WO 2004/009124 , and in particular in claims 1 to 31 of WO 2004/009124, as well as in the claims of WO 2005/068639, and therefore in particular in claims ia 29, which are explicitly incorporated herein by reference The preparation and use of another preferred embodiment of the second composition, wherein at least one second antigen comprises or is a nicotine, cotinine or nornicotine, has been described in the patent application WO 2004/009116 filed by the current assignee. This description is incorporated herein by reference. Other preferred embodiments of the second composition of the present invention are explicitly defined in the claims of WO 2004/009116 and particularly claims 1 to 50 of WO 2004/009116, which are explicitly incorporated herein by reference. In a preferred embodiment, at least one second antigen is formed from materials selected from the group comprising: (a) 6- (carboxymethylureido- (+) - nicotine (CMUNic); (b) trans-3 '-aminomethyl nicotine succinate (c) O-succinyl-3 '-hydroxymethyl-nicotine; (d) trans-4' -carboxycotinin; (e) N- [l-oxo-6- [(2S) -2- (3-pyridyl) - 1-pyrrolidinyl] hexyl] -β-alanine; (f) 4-oxo-4- [[6- [(5S) -2-oxo-5- (3-pyridinyl) -1-pyrrolidinyl]] hexyl] mino acid ) -butanoic (g) phenylmethyl ester of (2S) -2- (3-pyridinyl) -l-pyrrolidinbutanoic acid; (h) (2R) -2- (3-pyridinyl) -1-pyrrolidinbutanoic acid phenylmethyl ester; (i) cotinin 4'-carboxylic acid, N-succinyl-6-amino (+) - nicotine; (j) 6- (. sigma. -aminocapramido- (+) -nicotine; (k) 6- (. sigma. -aminocapramido- (+) - nicotine; (1) 3'-aminomethylnicotine; (m) 4'-aminomethyl nicotine; (n) 5'-aminomethylnicotine; (o) 5-aminonicotin; (p) 6-aminonicotine; (q) Sl- (b-aminoethyl) nicotinium chloride; (r) S-1- (b-aminoethyl) cotinium chloride; s) N-succinyl-6-amino- (+) nicotine.

In another preferred embodiment, the second composition comprises 0-succinyl-3 '-hydroxymethyl-nicotine. In another preferred embodiment, the second composition comprises 0-succinyl-trans-3 '-hydroxymethyl-nicotine. In another preferred embodiment, the second composition comprises 0-succinyl-3'-hydroxymethyl-nicotine conjugated to a virus-like particle of an RNA phage, preferably of a virus-like particle) and preferably with a virus-like particle. ) ß that comprises or preferably that it is composed of phage coat proteins RNA < 2ß. In a preferred embodiment, at least one second antigen comprises or is 0-succinyl-3 '-hydroxymethyl-nicotine. In another preferred embodiment, the second antigen comprises or is preferably O-succinyl-trans-3 '-hydroxymethyl-nicotine. In a preferred embodiment, the second antigen comprises or is preferably trans-3 '-hydroxymethyl-nicotine. In a preferred embodiment, the second binding site comprised by the second composition contains or is preferably an active group selected from the group comprising: (a) Amine; (b) Amida; (c) Carboxyl; (d) Carbonyl; (e) Sulfhydryl; (f) Hydroxyl; (g) Aldehyde; (h) Diazonium; (i) Allylhalogenide; (j) Hydrazine; (k) Vinyl; (1) Maleimida; (m) Succinimide; and (n) Hydrazide. In a preferred embodiment, the association of the first binding site with the second binding site by less a covalent bond is formed by the reaction of a 0-succinyl 0-succinyl-3 '-hydroxymethyl-nicotine entity with the first binding site. In another preferred embodiment, the association of the first binding site with the second binding site by at least one covalent bond is formed by an amide bond. In another preferred embodiment, the first binding site comprises or is preferably an amino group, preferably an amino group of lysine. In another preferred embodiment, the second binding site comprises or preferably is a carboxyl group. In another preferred embodiment, the association of the first binding site with the second binding site through at least one, preferably a covalent bond that is formed by the reaction of a 0-succinyl-3'-hydroxymethyl O-succinyl entity -nicotine with an amino group of lysine residue which is the first binding site. The second composition is preferably formed from mono- (1-methyl-2-pyridin-3-yl-pyrrolidin-3-ylmethyl) ester and succinic acid by reacting the carboxyl group of succinic acid with an amine group of phage RNA, preferably the amine group of lysine of phage RNA. A preferred way of carrying out the reaction is by activation of the carboxyl group with a carbodiimide, preferably EDC, even more preferably DCC. The activated carboxyl group can be directly linked covalently with phage RNA or can be converted to N-hydroxysuccinimide ester (NHS) by the addition of N-hydroxysuccinimide ester (NHS). The NHS ester is used directly after the reaction with the activated carboxyl group or is isolated and subsequently reacted with the phage RNA. Alternative reagents for active carboxyl groups are uronium salts such as HATU 2- (7-aza-lH-benzotriazol-l-yl) -1, 1,3,3-tetramethyluronium hexafluorophosphate) or HBTU (2- (lH) hexafluorophosphate -benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium). The mono- (l-methyl-2-pyridin-3-yl-pyrrolidin-3-ylmethyl) ester of succinic acid is activated by HATU or HBTA and the corresponding activated carboxyl groups are reacted directly with the amines. In a preferred embodiment, at least one second antigen is a GIP protein or a GIP fragment, wherein the second GIP protein or GIP fragment is different from the first GIP protein or first GIP fragment comprised in the first composition. In a preferred embodiment, the second GIP protein or GIP fragment comprises or consists of an amino acid sequence having different chemical modifications compared to the same amino acid sequence comprised or consisting of the first GIP protein or first GIP fragment. In a preferred embodiment, the second GIP protein or GIP fragment comprises or consists of an amino acid sequence different from the amino acid sequence comprised in or consisting of the first GIP protein or GIP fragment. In a preferred embodiment, the second GIP protein or GIP fragment comprises or consists of at least one different antigenic site compared to the antigenic sites comprised in or consisting of the first GIP protein or GIP fragment. In another preferred embodiment, the second GIP protein or GIP fragment comprises or consists of different antigenic sites compared to the antigenic sites comprised in or consisting of the first GIP protein or GIP fragment. In yet another preferred embodiment, the first GIP fragment comprises or consists of at least one antigenic site of the amino part of the GIP and the second GIP fragment comprises or consists of at least one antigenic site of the carboxyl part of the GIP. The carboxyl part of GIP refers to the last 18, preferably 15 amino acids of SEQ ID NO: 22 -26 or SEQ ID NO: 63. In a preferred embodiment, the first GIP fragment comprises or consists of an amino acid sequence as SEQ ID NO: 27 and the second first GIP fragment comprises or consists of an amino acid sequence of SEQ ID NO: 28. The characteristic and preferred embodiments, the elaboration and use of the different GIP proteins or GIP fragments have been described and defined in this invention.

In a preferred embodiment, the second GIP fragment comprises an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 27; (b) SEQ ID NO: 29; (c) SEQ ID NO: 32; (d) SEQ ID NO: 45; (e) SEQ ID NO: 28; (f) SEQ ID NO: 31; (g) SEQ ID NO: 44; (h) SEQ ID NO: 68 e (i) an amino acid sequence with at least 80%, preferably at least 85%, more preferably 90% and more preferably at least 95% identical with SEQ ID NO: 27-29, 31, 32, 44, 68 or 45. Except for the antigenic difference comprised in the first composition and the second composition, the characteristic and preferred embodiments, the preparation and use of the second composition is substantially the same as described and defined for the first. composition in this invention. In yet another aspect, the present invention provides a method for the treatment and / or prevention of obesity of an animal or human which comprises administering at least a first composition and at least a second composition of the kit in the same animal, preferably a cat or Domestic dog or a human. Administration of the second composition preferably occurs in the animal or human before, concomitantly with or subsequent to administration in the same animal or human of the first composition. In a preferred embodiment, both compositions are preferably administered in the same patient with a no more than two weeks apart, preferably no more than a week apart, more preferably no more than three days apart, even more preferably no more than 24 hours apart. In another preferred embodiment, both compositions are administered concomitantly in the same animal or human. The term concomitant, as used herein, refers to the fact that both compositions are mixed before being administered to an animal or human or that both compositions are administered in the same animal or human in succession. The term "in succession" as used herein, refers to that both compositions are administered in the same animal or human separately but with an interval not greater than 4 hours, preferably not greater than 2 hours, even more preferably not greater than one hour, even more preferably not more than half an hour and even more preferably not more than a 10 minute interval. Again the first preferred embodiment, the first composition and the second composition are administered by the same route, preferably the route is subcutaneous. In a preferred embodiment, the first composition and the second composition are administered concomitantly. In another aspect, the present invention provides a method for preventing and / or treating against obesity in an animal or human, which comprises administering the vaccine of the invention and a VLP-ghrelin vaccine, a VLP-nicotine vaccine or a VLP vaccine-a second GIP protein or a second GIP fragment in the same animal or human. The term "VLP-ghrelin vaccine" comprises the second composition of the invention, wherein at least one antigen is ghrelin or a ghrelin peptide. In a preferred embodiment, the "VLP-ghrelin vaccine" further comprises at least one adjuvant. Moreover, the "VLP-nicotine vaccine" comprises the second composition of the invention, wherein at least one antigen comprises a nicotine, a cotinine or a nornicotine. In a preferred embodiment, the "VLP-second GIP protein or second GIP fragment" vaccine comprises the second composition of the invention, wherein at least the antigen comprises or is a second GIP protein or GIP fragment wherein the second GIP protein or GIP fragment is different for the first GIP protein or GIP fragment. For reasons of simplicity, the vaccine of the invention and the VLP-ghrelin or VLP-nicotine or VLP-second GIP protein or second GIP fragment will hereinafter be referred to as "the two vaccines". Administration of the VLP-ghrelin vaccine or vaccine VLP-nicotine or the VLP-diflerent GIP protein vaccine or GIP fragment occurs in the animal or human before, concomitantly with or subsequent to the administration in the same animal or human of the vaccine of the invention. In a preferred embodiment, at least one antigen is a GIP protein or GIP fragment. The administration of the two vaccines in the same animal or human can trigger in an additive or preferably synergistic manner, the increase in efficacy compared to the administration of only one vaccine. In a preferred embodiment, at least one first composition and at least one second composition are kept separately in the kit. In another embodiment, at least one first composition and at least one second composition are mixed and maintained as a mixture in the kit. Therefore, in another aspect, the present invention provides a composition comprising (a) at least one virus-like first particle (VLP) and at least one second virus-like particle (VLP) each having at least one first site of binding and (b) at least a first antigen and at least a second antigen each having at least one second binding site, wherein at least the first antigen is a GIP protein or a GIP fragment and at least the second antigen comprises or is a molecule selected from the group consisting of: (i) a ghrelin or ghrelin peptide, (ii) a nicotine, a cotinine or nornicotine; (iii) a second GIP protein or second GIP fragment, wherein the second GIP protein or GIP fragment is different from the first GIP protein or GIP fragment and wherein the minus the first virus-like particle (VLP) and at least the first antigen bind by at least the first and at least the second binding site and wherein at least the second virus-like particle (VLP) and at least the second antigen binds by at least the first and by at least the second binding site. Therefore, the invention provides a vaccine composition comprising this composition. The vaccine composition may further comprise at least one adjuvant. Preferably, the vaccine composition is devoid of an adjuvant. The invention further provides an immunization method comprising administering the vaccine composition in an animal, preferably in a dog or a cat, preferably a domestic cat or a human. Moreover, the invention provides a method for the treatment and / or prevention of obesity which comprises administering the vaccine composition in an animal, preferably in a domestic or human dog or cat. Moreover, the invention provides a pharmaceutical composition comprising this composition and a pharmaceutically acceptable carrier. EXAMPLES The terms "prior art VLP" as well as the more specific terms "VLP < 2ß of the prior art", "VLP AP205 of the prior art" and the like, are they use within this section examples and refer to the VLPs obtained by the recombinant expression of E. coli and the subsequent purification as described in documents O 02/056905 and WO 04/007538. EXAMPLE 1 Chemical synthesis of GIP fragments 1-15, 1-10, 4-13, 16-30, 28-42, 31-42 and GIP protein 1-42 Mouse GIP fragments 1-15, 1-10, 4 -13, 16-30 and 31-42, including a GC or GC binding sequence fused to either the N- or the C-terminus of the GIP fragments (SEQ ID NO: 27.1 29, 32, 43, 44) they are synthesized chemically according to conventional procedures. The GIP 1-42 protein, including the GC binding sequence fused to the C-terminus (SEQ ID NO: 30) and the GIP fragment 28-42, including the CG linker fused to the N-terminus, are chemically synthesized according to conventional procedures . In order to improve the solubility of the N-erminal GIP fragments, coupled to the VLP?) ß of the prior art, the hydrophobic amino acid residues within the specific GIP fragment can be substituted. Polar, and hydrophilically charged amino acids such as lysine (lys) and aspartate (asp) are added or used to replace natural GIP residues. The GIP fragment 1-14, including a GIP fragment or DDC 1-14, including a KKC link sequence fused to the C-terminus of the GIP fragments are synthesized chemically according to conventional procedures. EXAMPLE 2 Coupling of GIP 1-15-GC, GIP 1-10-GC, GIP 4-13-GC and CG-GIP-31-42 with the VLP HBcAg, fr or? ß of the prior art A 2 ml solution of 2.0 mg / ml of VLP?) Of the prior art is reacted for 30 minutes in 20 mM Hepes, 150 mM NaCl pH 7.2, 114.4 μ? of a SMPH solution (Pierce) (from a stock solution 50 mM in DMSO) at 25 ° C. The solution of the reaction is subsequently subjected to the analysis 2 times for 2 hours against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C. The derivatized and dialyzed Qp VLP is subsequently used to couple either murine GIP 1-15-GC (SEQ ID NO: 60), GIP 1-10-GC (SEQ ID NO: 61), GIP 4-13- GC 8SEQ ID NO: 62) or murine CG-GIP 31-42 peptide. Briefly, 1 mi of VLP? derivatized (at a concentration of 2 mg / ml) is reacted with 286 μ? of 10 mM peptide solution for 2 hours at 20 ° C in 20 mM Hepes, 150 mM NaCl, pH 7.2. The coupling reactions are centrifuged at 13,000 rpm for 5 minutes and the supernatant is harvested and subjected to dialysis once for 2 hours and then overnight against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C. The covalent chemical coupling of GIP-1 peptides and 31-42 with the VLP? >; β is assessed by SDS-PAGE using 12% Un-PAGE gels (Invitrogen). The gels stained with Coomasie blue for the coupling reaction demonstrate the appearance of bands with molecular weights correspond to those predicted for GIP peptides covalently linked with < 2ß (Figure 1). The coupling bands correspond to 1, 2, 3 or 4 peptides coupled per subunit as indicated by the arrows. The appearance of additional bands compared to only VLP? > ß Derivatized, shows that GIP CG-1-15 and GIP 31-42-GC are covalently coupled with the VLP Qj3. The coupling efficiency [ie, mol C-GIP / mol monomer?) ß (total)] is estimated by densitometric analysis of SDS-PAGE stained with Coomasie blue, with results between 1.8-2.3 GIP fragments per monomer?) ß. Similar coupling efficiencies are observed for GIP CG-1-10 and GIP 4-13-GC that are covalently coupled with VLP Qj3 (no data shown). Coupling of GIP fragments with the VLP fr 1 ml of derivatized ffr VLP (at a concentration of 2 mg / ml) is reacted with 286 μ? of 10 mM GIP 1-15-GC, GIP 1-10-GC, GIP 4-13-GC or CG-GIP-31-42 for 2 hours at 20 ° C in 20 mM Hepes, 150 mM NaCl, pH 7.2. The coupling reactions are centrifuged at 13,000 rpm for 5 minutes and the supernatant is harvested and subjected to dialysis once for 2 hours and then overnight against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C.

Coupling of 6IP fragments with HBcAgl-185-Lys The structuring of HBcAgl-185-Lys, its expression and purification have been described substantially in EXAMPLE 2-5 of WO 03/040164. 1 ml of VLP HBcAgl-185-Lys (at a concentration of 2 mg / ml) is reacted with 286 μ? of 10 mM GIP 1-15-GC, GIP 1-10-GC, GIP 4-13-GC or CG-GIP-31-42 for 2 hours at 20 ° C in 20 mM Hepes, 150 mM NaCl, pH 7.2. The coupling reactions are centrifuged at 13,000 rpm for 5 minutes and the supernatant is collected and dialyzed once for two hours and then overnight against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C. EXAMPLE 3 Coupling of GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC and GIP 1-42-GC with the VLP? Μ of the prior art A solution of 2 ml of 2.0 mg / ml VLP 0_ß in 20 mM Hepes, 150 mM NaCl, pH 7.2 is reacted for 30 minutes with 114.4 μ? of a SMPH solution (Pierce) from 50 mM solution in stock dissolved in DMSO) at 25 ° C. The reaction is then subjected to dialysis twice for two hours against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C. The VLP?) ß subjected to dialysis and derivatization is subsequently used to couple murine GIP protein (SEQ ID NO: 30), murine CG-GIP 28-42, GIP 16-30-murine GC, GIP 1-14- KKC or GIP 1-14-DDC. In short, 1 ml of derivatized VLP? ß (at a concentration of 2 mg / ml) is made react with 286 μ? of 10 mM peptide solution for 2 hours at 20 ° C in 20 mM Hepes, 150 mM NaCl, pH 7.2. The coupling reactions are centrifuged at 13,000 rpm for 5 minutes and the supernatant is collected and subjected to dialysis once for 2 hours and then overnight against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C. The coupled products are then analyzed by SDS-PAGE using 12% Nu-PAGE gels (Invitrogen). Coupled products are visualized by Coomassie Brilliant Blue staining of the gels. EXAMPLE 4 Immunization of mice with GIP 1-15-GC, GIP 1-10-GC, GIP 4- 13-GC or CG-GIP 31-42 coupled to the VLP? Β of the prior art Adult male mice, C57BL are vaccinated / 6 (5 per group) either with murine GIP 1-15-GC, murine GIP 1-10-GC, murine GIP 4-13-GC or murine CG-GIP 31-42 coupled with VLP?) ß of the technique previous (obtained in EXAMPLE 2). 100 g of dialysed vaccine from each sample is diluted in PBS to obtain a volume of 200 μ? and injected subcutaneously (100 μ? on two ventral sides) on days 0, 14, 28 and 42. The vaccine is administered without adjuvant. As a control, a group of mice is injected with PBS or VLP?) ß. Mice are bled retro-biologically on days 0, 15, 28, 43, 56, 71, 91 and 105 and their serum is analyzed by ELISA as described in EXAMPLE 5. TABLE 1 shows the average titers of antibodies specific for GIP 1-15, specific for GIP 1-10, or specific for GIP 31-42. The results shown are the average of 10 mice per group. The ELISA titrations are expressed as serum dilutions that lead to the maximum mean OD in the ELISA assay. In mice immunized with GIP 1-15-GC-Q ^, GIP l-10-GC-Q or < 2ß-CG-GIP 31-42, the average titers of approximately 1: 300 000, 1: 330 000 and 1:82 000, respectively, are reached on day 56 (TABLE 1). Specific titers for GIP of mice immunized with GIP 4-13-GC-? Β are approximately 10 times lower than the titers specific for GIP 1-15 (data not shown). Pre-immunized sera or sera from mice immunized with PBS or VLP < 2ß showed no sensitization against either GIP peptide. The maximum mean DO titration was less than 100, which is considered to be below the assay discriminant value. This clearly demonstrates that the GIP-VLP conjugate is able to induce a high titre of antibodies against GIP fragments although it is a selfprotein. Antibody titers determined against GIP fragments coupled to AR asa are similar to those obtained against the GIP protein (no data shown). This indicates that the antibodies sensitized against GIP fragments are able to recognize the GIP protein.

TABLE 1. Average anti-GIP titre specific for IgG (expressed as dilution factor) in mice immunized on day 0, 14, 28 and 42 with GIP 1-15-GC-QJ3, GIP 1-10 -GC-Qp or Q-GG-GIP 31--42, respectively.

Days after the first immunization Immunization 14 28 42 56 70 Qb-GIP 1-15 75 463 263 598 266 839 306 169 239 527 Qb-GIP 1-10 88 057 331 444 270 214 336 882 307 235 Qb-GIP 31-42 15 480 47 974 70 815 82 066 70 458 PBS or VLP Qb 100 100 100 100 100 EXAMPLE 5 Detection of GIP-Specific and Ghrelin-Specific Antibodies in an ELISA Assay First, murine GIP 1-15-GC or murine GC-GIP 31-42 is coupled with RNase (SIGMA)-SPDP (SIGMA) as shown in FIG. described in the next paragraph. 5 mg / ml R asa and 0.2 mM SPDP (final concentration) are incubated for one hour at room temperature. The RNase-SPDP solution is purified on a PD10 column (Amersham). After purification, 10 mM EDTA and 1 mM peptide are added to the RNase-SDPD solution. The coupling efficiency was determined by measuring OD at 343 nm. ELISA plates are coated (96 wells MAXIsorp) coated with murine GIP 1-15-GC coupled with RNase or murine CG-GIP 31-42 at a concentration of 10 μ / 1 in a coating buffer (0.1 M NaHCO3, pH 9.6), overnight 4 ° C. Alternatively, ELISA plates are coated with 2.5 μg / ml porcine GIP protein (Bachem). After washing the plates in wash buffer (PBS-0.05% Tween), the plates are blocked with wash buffer (2% BSA-PBS-Tween 20 solution) for 2 hours at 37 ° C and then washed again and incubate with mouse serum diluted serially. As a control, a pre-immune serum from the same mouse is also analyzed. The plates are incubated at room temperature for two hours. After another wash, the bound antibodies are detected by a goat anti-mouse IgG antibody specific for Fe and labeled with HRPO (Jackson Immunoresearch) and incubated for one hour at room temperature. After further washing, the plates are developed with an OPD solution (one OPD tablet, 25 ul OPD buffer and 8ul H202) for 6 minutes and the reaction is terminated with a 5% H2SO4 solution. The plates are read at 450 nm in an ELISA reader (Biorad Benchmark). ELISA titrations are expressed as serum dilutions that lead to a mean maximum OD in an ELISA assay. In order to measure ghrelin-specific antibodies from sera from mice immunized with Ghrelin 24-31GC coupled with VLP?) ß, a similar method was used for that described above. The only difference is that the ELISA plates are coat with 20 ug / ml ghrelin protein (Bachem). EXAMPLE 6 Efficacy experiments with GIP 1-15-GC-. { J $ # GIP l-10-GC-Qji or? ß-CG-GIP 31-42 coupled to VLP?, ß of the prior art in an experimental model of obesity in an animal induced by diet Adult male mice C57BL / 6 (5 per group) with comparable initial weights (22.7 - 23.1 g) were vaccinated, as described in EXAMPLE 4 with either GIP 1-15-murine GC, GIP 1-10-GC or CG-GIP 31-42 of murine coupled to the VLP < 2ß, obtained in EXAMPLE 2. As a control, the mice were injected with VLP?) ß. The mice obtained a diet high in fat (35% fat by weight, 60% as energy) after the first injection, in order to facilitate the development of diet-induced obesity. Water and food ad limitum was administered. The body weights of individual animals were monitored at regular intervals for a period of time approximately 4 months after the injection. As shown in FIGURE 2A, mice immunized with GIP 1-15-GC or CG-GIP 31-42 coupled with VLP?) ß obtained less weight gain during the course of the experiment than the control animals, which had been injected with VLP?) ß. In fact, 112 days after the first immunization the control animals had increased their weight around 110% while mice immunized with GIP 1-15-GC-QP and CG-GIP 31-42 -?) ß had only increased their weight by 80% and 82%, respectively. Therefore, both vaccinated groups clearly showed a lower weight gain compared to the control groups. Results comparable to those achieved with GIP 1-15-GC of VLP?) Were obtained with GIP l-lO-GC-? ß (data not shown). To further evaluate the effect of vaccination against GIP, changes in body fat were quantified by dual-energy X-ray absorptiometry (DEXA). The DEXA analyzes were carried out using a high resolution densitometer PIXIMUS series (pixels 0.18 x 0.18 mm, GE Medical Systems). 142 days after the first injection, DEXA analyzes were carried out. Mice immunized with GIP 1-15-GC or CG-GIP 31-42 coupled to VLP? ß showed a reduction in body fat by approximately 26% and 22%, respectively (IGURA 2B) compared to control animals VLP <2ß. Similar results were obtained with GIP 1-IO-GC-Q coupled to VLP?) ß (data not shown). No differences in lean body mass were observed between the control and vaccinated groups, which clearly indicates that the reduction in body weight was due to the reduction in body fat. Taken together, these results clearly demonstrate that the GIP-VLP conjugate is capable of reduce the gain of body weight and the accumulation of body fat. EXAMPLE 7 Experiments on safety with GIP l-15-GC- < $, GIP 1-10-GC-? ß or?, ß-CG-GIP 31-42 coupled with VLP?, ß of the prior art in an experimental model in animal-induced diet obesity To evaluate possible side effects of the vaccination against GIP, blood glucose, fructosamine and triglycerides, the levels were quantified in vaccinated animals. Briefly, adult male C57BL / 6 mice (five per group) were vaccinated as described in EXAMPLE 4 with GIP 1-15-GC or CG-GIP 31-42 coupled to VLP? ß as obtained in EXAMPLE 2. As a control, the lratons were injected with PBS or VLP QJ. Over the course of 4 days, separated by two days of difference between 91 and 102 days after the first immunization, the mice were bled twice 'during the morning (9.00 hours) and twice in the afternoon (15.00 hours) and determined the blood glucose level using the Glucotrend blood glucose meter (Roche). During this period, the mice had free access to water and food. The averages of the respective morning and afternoon readings are shown in TABLE 4. No significant difference was observed in terms of levels of blood glucose between animals injected with PBS and those vaccinated. Similar results were observed in mice vaccinated with GIP 1-10-GC-Q (no data shown). In the same animals, plasma triglyceride levels were determined 120 days after the first injection. In summary, after a period of 12 hours of fasting, blood samples were obtained from mice immunized with GIP 1-15-GC or CG-GIP 31-42 coupled to VLP?) And control mice injected with PBS. Triglyceride levels of plasma samples were determined by enzymatic assays with an automated Olympus AU400 laboratory workstation. The average values for each group and the standard deviations (n = 5) are shown in TABLE 4. No significant differences were observed between the control and vaccinated animals. Furthermore, fructosamine levels were determined in the same animals at various time intervals after immunization. Fructosamine levels reflect the total amount of glycosylated protein in the circulation and if the fructosamine levels are elevated it indicates an increase in glycemia. Since fructosamine in blood circulation has a period of three weeks, the measurement of fructosamine provides a retrospective glycemic picture. In summary, after a fasting period of 12 hours, blood samples are obtained at various intervals of time from of mice immunized with GIP 1-15-GC-C $ or CG-GIP 31-42-? ß and from control mice injected with VLP < 2ß or PBS. Then the fructosamine levels are determined from plasma samples. The average values for each group and the standard deviations (n = 5) are shown in TABLE 5. In general, there were no significant differences observed between the control and vaccinated animals. Despite this, two time points significantly different for the group vaccinated with GIP 1-15-T? 0ß or CG-GIP 31-42-? ß (days 55 and 69, days 27 and 69, respectively), the overall values they were in the normal range for fructosamine, approximately 200-400 μmol / L. Fructosamine levels exceeding 400 | imol / L are considered a risk for the development of diabetes. Therefore, vaccination against GIP does not induce a hyperglycemic state in mice immunized with GIP 1-15-GC or CG-GIP 31-42 coupled with VLP?). In conclusion, no significant differences were observed in blood glucose levels, fasting levels of triglycerides and fructosamine between the vaccinated group (GIP 1-15-GC or CG-GIP 31-42 coupled with VLP?) ß) and the control group (animals injected with VLP?) ß or PBS) despite the presence of anti-GIP antibodies in the vaccinated mouse (TABLES 2 and 3).

TABLE 2 Mean Triglyceride Levels During Fasting and Average Blood Glucose Levels During Morning and Afternoon in Inco Mice Per Group Immunized with GIP 1-15-GC-C $ or CG-GIP 31-42 TABLE 3 Average fructosamine levels during fasting mice per group with GIP l-15-GC-Qp or CG-GIP 31 EXAMPLE 8 Immunization of mice with GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC and GIP 1-42-GC protein coupled to the VLP?, Β of the Prior art C57BL / 6 male or female adult mice were vaccinated with either GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, murine GIP 1-14-DDC or GIP protein 1-42 -GC coupled to VLP? > ß, obtained in EXAMPLE 3. In summary, 100 g of vaccine subjected to dialysis of each sample are diluted in PBS at a volume of 200 μ? and injected subcutaneously (100 μ? on two ventral sides) on days 0, 14, 28 and 42 and subsequently as required. The vaccine is administered with or without adjuvant. As a control, a group of mice are immunized with VLP? ß or injected with PBS with or without adjuvant. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. The GIP-specific antibodies are then quantified by ELISA as described in EXAMPLE 5. EXAMPLE 9 Efficacy experiments with GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14- KKC, GIP 1-14-DDC and GIP protein 1-42-GC coupled to VLP?, ß in an animal model of diet-induced obesity Female adult mice or C57BL / 6 male with compared initial weights are vaccinated as described at EXAMPLE 8 either with GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC of murine or GIP protein 1-42-GC coupled to VLP?) ß, obtained in EXAMPLE 3. as a control, mice are immunized with VLP?) ß or injected with PBS. The mice subsequently receive a booster vaccine with GIP-specific antibody titers that significantly decline during the experiment. All mice are fed a diet high in fat (35% fat by weight, 60% as energy) to facilitate the development of diet-induced obesity. Water and food ad limitum is administered. Body weights are monitored at regular intervals. In addition to body fat mass, blood glucose levels and plasma fructosamine and triglyceride levels are determined at different intervals as described in EXAMPLE 7. EXAMPLE 10 Efficacy experiments with GIP 1-15-GC, GIP 1-10 -GC, GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC of murine and protein GIP 1-42-GC coupled to the VLP? ß of the prior art in a model of obesity in a genetic animal Adult male or female mice, C57BL / 6 ob / ob are vaccinated as described in EXAMPLE 8 with either GIP 1-15-GC, GIP 1 -10-GC, GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, murine GIP 1-14-DDC and GIP protein 1-42-GC coupled to the VLP?) ß of the prior art, obtained in EXAMPLE 2 or 3. As controls, the mice are immunized with VLP? ß or are injected with PBS. The mice subsequently receive a booster vaccine if the GIP-specific antibody titers declined significantly for a period of time in the experiment. Mice are fed a conventional diet (consisting of 4-10% fat by weight), ad limitum and have free access to water. Body weights are monitored at regular intervals. In addition to the body fat mass, fructosamine and plasma triglyceride levels and blood glucose levels are determined at different intervals (as described in EXAMPLE 7). EXAMPLE 11 Efficacy experiments with GIP 1-15-GC, GIP 1-10-GC and CG-GIP 31-42 coupled to VLP oj3 in an animal model of obesity induced by therapeutic diet Adult male C57BL / 6 mice (10 per group) are fed a high-fat diet, ad limitum, for approximately 14 weeks until they become obese (weights> 42g). Then two experimental groups of similar average initial weight are made. The averages of both groups differ only in < 0.1 g at the beginning of the experiment. After grouping, the mice (10 per group) vaccinate as described in EXAMPLE 4, with GIP 1-15-GC, GIP 1-10-GC and CG-GIP 31-42 of murine coupled to the VLP? ß of the prior art, obtained in EXAMPLE 2. As a control , the mice are immunized with VLP? ß. The mice subsequently receive a booster vaccine if the GIP specific antibody titers declined significantly during the experiment. Mice are bled retroorbitally on day 0, 14, 28, 42, 56 and subsequently at monthly intervals. The sera are analyzed for GIP-specific antibodies in an ELISA specific for GIP as described in EXAMPLE 5. The titers of antibodies specific for GIP induced by vaccination with GIP 1-15-GC, GIP 1-10-GC or CG- GIP 31-42 coupled to the VLP?) ß of the prior art are comparable to those shown in TABLE 1. the body weights are monitored at regular intervals. In addition to body fat mass, blood glucose levels and plasma triglyceride levels are determined at different intervals as described in EXAMPLE 7. As shown in TABLE 4, mice immunized with GIP 1-15-GC , GIP 1-10-GC or CG-GIP 31-42 coupled to the VLP?) ß increased less weight during the course of the experiment than the control animals that had been injected with VLP? ß. By day 98 after the first immunization, the control animals had increased their weight approximately 8.6 g (2%) while mice immunized with GIP 1-15-T? 0ß, GIP l-lO-GC-? ß or CG-GIP 31-42 VLP? ß had decreased their weight by 0.5 g ( 3.0%), 0.6 g (-3.0%) and 0.2 g (-0.5%), respectively. Therefore, all accumulated groups clearly demonstrate a lower weight gain compared to control groups, in a therapeutic setting. TABLE 4 Change in average body weight (expressed as a percentage) of ten mice per group immunized with GIP l-15-GC-Q, GIP l-lO-GC-? ßO CG-GIP 31-42 VLP?) ß in a 98 day period EXAMPLE 12 Efficacy experiments with GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC from murine and GIP protein 1-42-GC Coupled to VLP?, ß in an animal model of obesity induced by therapeutic diet Adult male or female C57BL / 6 mice are fed a high-fat diet, ad limitum, for approximately 17-24 weeks or until they have become obese (weights> 45g). The mice are then grouped so that the distribution of the initial weights and the average initial weight is similar for all groups. Mice are vaccinated as described in EXAMPLE 8, either with GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC of murine or GIP protein 1-42-GC coupled to the VLP QJ3, obtained in example 3. As controls the mice are immunized with VLP Qj3 or injected with PBS. The mice also receive a booster vaccine if the GIP-specific antibody titers begin to decline. The mice are bled retroorbitally on day 0, 14, 28, 42, 56 and 70 and then at monthly intervals. The sera are analyzed to determine antibodies specific for GIP by ELISA as described in EXAMPLE 5. The body weights are monitored at regular intervals. In addition to the body fat mass, the levels of plasma amino acids and triglycerides and blood glucose levels are determined at different intervals, as described in EXAMPLE 7.

EXAMPLE 13 Preparation of VLP?, ß of the invention by disassembly / reassembly in the presence of different polyanionic macromolecules resulting in reassembled VLP 0_ß (A) VLP Disassembly < 2β of the prior art 45 mg of VLP?) ß of the prior art (2.5 mg / ml, as determined by Bradford analysis) in PBS (20 mM phosphate, 150 mM NaCl, pH 7.5) purified from lysate of E. coli , it is reduced with 10 mM DTT for 15 minutes at room temperature with stirring. Magnesium chloride is then added at 0.7 M final concentration and incubation is continued for 15 minutes at room temperature with stirring, which leads to the precipitation of encapsulated host cell RNA. The solution is centrifuged for 10 minutes at 4000 rpm at 4 ° C (Eppendorf 5810 R, in a fixed-angle rotor A-4-62 used in the following steps) in order to remove the precipitated RNA from the solution. The supernatant, which contains the cover protein γ) dimer and released, is used for chromatographic purification steps. (B) Purification of the cover protein?) ß by cation exchange chromatography and by size exclusion chromatography. The supernatant of the disassembly reaction, which contains the dimeric coat protein, cell protein host and residual host cell RNA, diluted 1:15 in water to adjust its conductivity below 10mS / cm and loaded on an SP-Sepharose FF column (xkl6 / 20, 6 ml, Amewrsham Bioscience). The column is equilibrated in advance with 20 mM sodium phosphate buffer at pH 7. The elution of the bound coat protein is achieved by a gradient in steps at 20 mM sodium phosphate / 500 mM sodium chloride and the protein is collects in a fractional volume of approximately 25 ml. Chromatography is carried out at room temperature with a flow rate of 5 ml / min and the absorbance is monitored at 260 nm and 280 nm. In the second step, the isolated ββ coat protein (the fraction eluted from the cation exchange column) is loaded (in two series) onto a Sephacryl S-100 HR column (xk26 / 60, 320 ml, Amersham Bioscience), balanced with 20 mM sodium phosphate / 250 mM sodium chloride; pH 6.5. Chromatography is carried out at room temperature with a flow rate of 2.5 ml / min and the absorbance is monitored at 260 nm and 280 nm. Fractions of 5 ml are collected. (Cl) Re-assembly of VLP?) ß by dialysis The cover protein?) ß purified (2.2 mg / ml in 20 mM sodium phosphate pH 6.5), a polyanionic macromolecule (2 mg / ml in water), urea (7.2 M in water) and DTT (0.5 M in water) are mixed at the final concentrations of 1.4 mg / ml cover protein, 0.14 mg / ml of the polyanionic macromolecule respective, 1 M of urea and 2.5 mM of DTT. The mixtures (each of 1 ml) are subjected to dialysis for two days at 5 ° C in 20 mM TrisHCl, 150 mM NaCl pH 8, using membranes excluding 3.5 kDa. The polyanionic macromolecules are: polygalacturonic acid (25000 - 50000, Fluka), dextran sulfate (molecular weights of 5000 and 10000, Sigma), poly-L-aspartic acid (molecular weights 11000 and 33400, Sigma), poly-L-acid glutamic (molecular weights 3000, 13600 and 84600, Sigma) and yeast tRNA for baking and wheat germ. (C2) Re-assembly of VLP?) ß by diafiltration 33 ml of purified? -coating protein (1.5 mg / ml in 20 mM sodium phosphate pH 6.5, 250 mM NaCl) are mixed with water and urea (7.2 M in water) , NaCl (5 M in water) and poly-L-glutamic acid (2 mg / ml in water, molecular weight: 84600). The volume of the mixture is 50 ml and the final concentrations of the components are 1 mg / ml cover protein, 300 mM NaCl, 1.0 M urea and 0.2 mg / ml poly-L-glutamic acid. The mixture is then diafiltered at room temperature, against 500 ml of 20 mM TrisHCl pH 8, 50 mM NaCl, applying a cross flow rate of 10 ml / min and an infiltrate flow rate of 2.5 ml / min, in an apparatus of tangential flow filtration using a Pellicon XL membrane cartridge (Biomax 5K, Millipore).

EXAMPLE 14 In vitro assembly of VLP AP205 (A) Purification of AP205 cover protein Disassembly: 20 ml of a VLP AP205 solution (in 1.6 mg / ml PBS, purified from an extract of E. coli) is mixed with 0.2 ml of 0.5 M DTT and incubated for 30 minutes at room temperature. 5 ml of 5M NaCl are added and the mixture is incubated for 15 minutes at 60 ° C causing the precipitation of the coat proteins reduced with DTT. The cloudy mixture is centrifuged (Sorvall SS34 rotor, 1000 g, 10 min, 20 ° C) and the supernatant is discarded and the pellet is dispersed in 20 ml of 1 M urea / 20 mM sodium treatment pH 3.2. After stirring for 30 minutes at room temperature, the dispersion is adjusted to pH 6.5 by the addition of 1.5 M Na2HP04 and then centrifuged (Sorvall SS34 rotor, 10000 g, 10 min, 20 ° C) to obtain the supernatant containing the Dimeric cover protein. Cation exchange chromatography: The supernatant (see above) is diluted with 20 ml of water to adjust a conductivity of approximately 5 mS / cm. The resulting solution is loaded onto a 6 ml SP Sepharose FF column (Amersham Bioscience) which is pre-equilibrated with 20 mM sodium phosphate buffer pH 6.5. After loading, the column is washed with 48 ml of 20 mM sodium phosphate pH 6.5. After loading, the column is washed with 48 ml of 20 mM Sodium phosphate buffer pH 6.5 followed by elution of the bound coat protein by a linear gradient at 1 M NaCl with respect to 20 column volumes. The main peak fractions are pooled and analyzed by SDS-PAGE and UV spectroscopy. According to SDS-PAGE, the isolated coat protein is essentially pure from other protein contaminants. According to UV spectroscopy, the protein concentration is 0.6 mg / ml (total amount 12 mg), considering that an A280 unit reflects 1.01 mg / ml AP205 cover protein. Moreover, the value of A280 (0.5999) with respect to the value of A260 (0.291) is 2, which indicates that the preparation is essentially free of nucleic acids. (B) Assembly of VLP AP205 The assembly in the absence of any polyanionic macromolecule: The protein fraction eluted in the previous paragraph is diafiltered and concentrated by TFF at a protein concentration of 1 mg / ml in 20 mM sodium phosphate pH 6.5. 500 μ? of that solution are mixed with 50 μ? of a 5 M NaCl solution and incubate for 48 hours at room temperature. The formation of VLPs reassembled in the mixture is demonstrated by non-reducing SDS-PAGE and by HPLC by size exclusion. A G5000 P XL column in TSK gel (Tosoh Bioscience), equilibrated with 20 mM sodium phosphate, 150 mM NaCl pH 7.2, is used for HPLC analysis.

Assembly in the presence of polyglutamic acid: 375 μ? of purified AP205 cover protein (1 mg / ml in 20 mM sodium phosphate at pH 6.5) is mixed with 50 μ? of a solution in the presence of NaCl (5 M in water), 50 μ? of solution in the presence of polyglutamic acid (2 mg / ml in water, molecular weight: 84600, Sigma) and 25 μ? of water. The mixture is incubated glove 48 hours at room temperature. The formation of re-assembled VLP in the mixture is demonstrated by non-reducing SDS-PAGE and by size exclusion HPLC. The coat protein in the mixture was almost completely incorporated in the VLPs, which shows a higher assembly efficiency than the AP205 cover protein assembled in the absence of any polyanionic macromolecule. EXAMPLE 15 GIP coupling fragments GIP 1-10, 4-13, 1-15, 31-42, 16-30, 28-42, 1-14-KKC, GIP 1-14-DDC and GIP protein 1-42 in VLP? ß or re-assembled VLP AP205 To a 2 ml solution of 2.0 mg / ml VLP?) Reassembled ß (obtained in EXAMPLE 13) in 20 mM Hepes, 150 mM NaCl, pH 7.2 is reacted for 30 minutes with 114.4 μ? of a SMPH solution (Pierce) (from a stock solution of 50 mM dissolved in DMSO) at 25 ° C. The reaction solution is subsequently dialyzed twice for 2 hours against 2 liters of 20 mM Hepes, 150 mM NaCl, H 7.2 at 4 ° C. Re-assembled, derivatized and dialyzed VLP?) ß is subsequently used to couple murine GIP 1-15-GC, murine CG-GIP 31-42 peptide, murine GIP 1-10-GC, murine GIP 4-13-GC, GIP 16-30-GC of murine, CG-GIP 28-42 of murine, GIP 1-14-KKC of murine, GIP 1-14-DDC of murine or protein GIP 1-42-GC of murine. In brief, 1 ml of VLP?) ß reassembled and derivatized (at a concentration of 2 mg / ml) is reacted with 286 μ? of 10 mM peptide solution for two hours at 20 ° C in 20 mM Hepes, 150 mM NaCl, pH 7.2. The coupling reactions are then centrifuged at 13,000 rpm for 5 minutes and the supernatant is collected and dialyzed once for two hours and then overnight against 2 liters of 20 mM Hepes, 150 mM NaCl, pH 7.2 at 4 ° C. . The 2 ml solution of 2.0 mg / ml of reassembled VLP AP205 (obtained in EXAMPLE 14) in 20 mM Hepes, 150 mM NaCl, pH 7.2 is first derivatized by SMPH under identical or similar conditions as described for VLP?). reassembled Re-assembled and derivatized AP205 VLP is reacted with GIP fragments as described in the latter under similar and identical conditions.

EXAMPLE 16 Efficacy and immunization experiments with GIP 1-10-GC, GIP 1-15-GC, GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14- KKC, GIP 1-14-DDC, CG-GIP 31-42 and GIP 1-42-GC protein coupled to reassembled ß-VLP or re-assembled AP205 VLP in an experimental animal model of obesity induced by therapeutic diet Adult male or female mice, C57BL / 6 are fed a high-fat diet, ad limitum, for approximately 17-24 weeks or until the mice have become obese (weights >; 45g). The mice are then grouped so that the distribution of the initial weights and the average initial weight is similar for all groups. The mice are vaccinated as described in EXAMPLE 8 with either GIP 1-10-GC, GIP 1-15-GC, GIP 4-13-GC, GIP 16-30-GC, CG-GIP 28-42, GIP 1-14-KKC, GIP 1-14-DDC, murine CG-GIP 31-42 or GIP 1-42-GC coupled to the VLP?) Reassembled ß obtained in EXAMPLE 15. The control mice are immunized with VLP?) ß reassembled (obtained in EXAMPLE 13) or injected with PBS. 100 μg of control protein or dialyzed vaccine is diluted in a volume of 200 ml of PBS and injected subcutaneously (100 μm on two ventral sides) with or without adjuvant on day zero. The mice are given a booster vaccine with the corresponding formulation on day 14, 28 and 42. The mice are given another immunization of reinforcement if the specific antibody titers for GIP begin to decline. The mice are bled retroorbitally on day 0, 14, 28, 42, 56, 70 and then subsequently at regular intervals. The sera are analyzed for antibodies specific for GIP by ELISA as described in EXAMPLE 5. The body weights are monitored at regular intervals. In addition to body fat mass, fructosamine levels and plasma triglycerides and blood glucose levels at different intervals as described in EXAMPLE 7. determined Similar conditions apply, similar or identical to analyze the effectiveness of AP205 VLP reassembled coupled to GIP fragments or GIP protein in the experimental mouse model of obesity induced by therapeutic diet. EXAMPLE 17 Coupling ghrelin 24-31-GC to VLP ß prior art The ghrelin peptide Ghrel24-31-GC (GSSFLSPEGC SEQ ID NO: 39)?, Comprising another residue C-terminal cysteine for coupling to VLP , it is synthesized chemically and used for the chemical coupling in the? ß as described below. A solution of 5 ml of 140? of VLP?) ß of the prior art in 20 mM Hepes, 150 mM NaCl pH 7.4 is made react for 30 minutes with 108 μ? 65 mM solution of SMPH (Pierce) in H20 at 25 ° C on a rng shaker. The reaction solution is subsequently dialyzed twice for 2 hours against 5 liters of 20 mM Hepes, 150 mM NaCl pH 7.2 at 4 ° C. 100 μ? of the dialyzed reaction mixture are reacted with either 28.6 μ? of a solution in existence (in DMSO) of the peptide ghrelin (1:10 peptide /? ß capsid protein ratio). The coupling reaction is carried out for two hours at 15 ° C in a water bath. The reaction mixture is centrifuged at 13,000 rpm for 5 minutes and the supernatant is ccted and dialyzed once for two hours and then overnight against 2 x 5 L of 20 mM Hepes, 150 mM NaCl pH 7.2 at 4 ° C. The coupling reaction is analyzed on 16% SDS-PAGE gels by reducing conditions. The gels are stained with Coomasie Brilliant Blue. EXAMPLE 18 Simultaneous immunization of mice with 6IP 1-15-GC coupled to VLP? ß and Ghrel24-31-GC coupled to VLP? ß Female or male adult mice, C57BL / 6, are simultaneously vaccinated with GIP 1-15-GC from murine coupled to VLP?) ß (obtained from EXAMPLE 2) and murine ghrel24-31-GC coupled to VLP < 2ß (obtained from EXAMPLE 17). 100 μg of each dialysed vaccine are combined and diluted in PBS to a volume of 200 μ? and injected subcutaneously (100 μ? on two ventral sides) on days 0, 14, 28 and 42 and subsequently as required. The combination of two vaccines is administered with or without adjuvant. As a control, a group of mice is immunized with only VLP?) ß or injected with PB, with or without adjuvant. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. Their serum is analyzed for anti-GIP and anti-ghrelin antibodies by ELISA, as described in EXAMPLE 5. Similar, analogous or identical experimental conditions are applied to immunize male or female adult mice, C57BL / 6 simultaneously with GIP 1-15 -GC coupled or VLP? ß reassembled and ghrel24-31-GC coupled to the reassembled VLP Qj3. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals and their serum is analyzed for anti-GIP and anti-ghrelin antibodies using Ghrelin-specific or GIP-specific ELISA, respectively, as described in Example 5. Example 19 Experiments with simultaneous immunization efficacy d® GIP 1-15-GIC coupled to VLP? ß and Ghrel24-31-GC coupled to VLP?, ß in a model of obesity induced by diet adult male or female mice , C57BL / 6, with comparable initial weights are vaccinated with either murine GIP 1-15-GC coupled to the VLP? ß of the prior art or as the murine ghrel24-31-GC reassembled and coupled to VLP <2ß of the prior art, as described in EXAMPLE 18. As a control, the mice are immunized with VLP? ß of the prior art or VLP? ß reassembled only or injected with PBS. The mice are subsequently given a booster immunization if the titration of Ghrelin-specific or GIP-specific antibodies declines significantly during the experiment. All mice are given a high-fat diet (35% fat by weight, 60% as energy) to facilitate the development of diet-induced obesity. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. His serum is analyzed for anti-GIP and anti-ghrelin antibodies by ELISA, as described in EXAMPLE 5. Water and food ad limitum are administered. Body weights are monitored at regular intervals. In addition, body fat mass, blood glucose levels and plasma triglyceride levels are determined at different intervals as described in EXAMPLE 7. EXAMPLE 20 Efficacy experiments with simultaneous immunization of GIP 1-15-GC coupled to VLP ?, ß in a genetic model of animal obesity Adult male or female mice, C57BL / 6, are vaccinated simultaneously with GIP 1-15-GC of murine coupled to the VLP?) ß of the prior art or reassembled VLP? ß and ghrelin 24-31-GC coupled to the VLP?) ß of the prior art or VLP Qj3 reassembled as described in EXAMPLE 18. As controls, the mice are immunized with VLP? ß of the prior art, VLP?) ß is reassembled or injected with PBS. Subsequently, the mice are given a booster immunization if the GIP-specific or ghrelin-specific antibody titers declined significantly during the period of the experiment. The mice are fed a conventional diet. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. The serum is analyzed for anti-GIP and anti-ghrelin by ELISA as described in EXAMPLE 5. The body weights are monitored at regular intervals. In addition to the body fat mass, the plasma triglyceride levels and blood glucose levels are determined at different intervals as described in EXAMPLE 7. EXAMPLE 21 Efficacy experiments with simultaneous immunization of GIP 1-15-GC coupled to the VLP ? ß and Ghrelin 24-31-GC coupled to VLP?, ß in an animal model of obesity induced by therapeutic diet Adult male or female mice, C57BL / 6 they feed on a high-fat diet, ad limitum, for approximately 17-24 weeks or until they have become obese (weight> 45g). The mice are then grouped so that the distribution of the initial weights and the average initial weight is similar for all groups. Mice are simultaneously vaccinated with murine GIP 1-15-GC coupled to the VLP?) Of the prior art or the VLP?) Reassembled as with ghrelin 24-31-GC coupled to the VLP?) ß of the previous technique or the VLP < 2β reassembled as described in EXAMPLE 18. The control mice are immunized with the VLP?) ß of the prior art or the VLP?) ß reassembled or injected with PBS. Subsequently, the mice are given a booster immunization if the GIP-specific or ghrelin-specific antibody titers declined significantly during the experiment. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. The serum is analyzed for anti-GIP and anti-ghrelin antibodies by ELISA as described in EXAMPLE 5. Body weights and body fat composition are monitored in individual mice after immunization. In addition, triglyceride and blood glucose levels are monitored at different intervals as described in EXAMPLE 7.

EXAMPLE 22 Simultaneous Immunization of Mice with GIP 1-15-GC Coupled to VLP? ß and CG-GIP 31-42 Coupled to VLP? ß Adult mice, females or males, C57BL / 6 are simultaneously vaccinated with GIP 1- 15-murine GC coupled to the VLP ß and CG-GIP 31-42 of murine coupled to the VLP?) ß (both obtained from EXAMPLE 2). 100 μg of each dialysed vaccine are combined and diluted in PBS to a volume of 200 μ? and injected subcutaneously (100 μ? on two ventral sides) on days 0, 14, 28 and 42 and subsequently as required. The combination of two vaccines without adjuvant is administered. As a control, the mice are immunized with VLP Qj3 alone or injected with PBS. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. Your serum is analyzed for antibodies specific for anti-GIP by ELISA by coating the full-length mouse GIP protein on the plates, as described in EXAMPLE 5. The GIP-specific antibody titers are comparable to those shown in TABLE 1. EXAMPLE 23 Efficacy experiments with simultaneous immunization of GIP 1-15-GC CG-GIP 31-42 coupled to the VLP?, ß and CG-GIP 31-42 CG-GIP 31-42 coupled to the VLP? , ß in a model of diet-induced obesity Adult mice, females or males, C57BL / 6 with comparable initial weights are vaccinated with both GIP 1-15- Murine GC coupled to the VLP?) ß as with the CG-GIP 31-42 coupled to the VLP?) ß, as described in EXAMPLE 22. As a control, the mice are immunized with the VLP?) ß of the prior art and then injected with PBS. The mice are subsequently given a booster immunization if the GIP-specific antibody titers decline significantly during the experiment. All mice are placed on a high-fat diet (35% fat by weight, 60% as energy) to facilitate the development of diet-induced obesity. Mice are bled retroorbitally on day 0, 14, 28, 42 and subsequently at regular intervals. Your serum is analyzed for anti-GIP antibodies by ELISA, as described in EXAMPLE 5. The specific antibody titers for GIP are comparable to those shown in TABLE 1. Water and food ad limitum is administered. Body weights are monitored at regular intervals. In addition to the body fat mass, plasma fructosamine and triglyceride levels and blood glucose levels are determined at different intervals as described in EXAMPLE 7. The average body weight gain in mice immunized simultaneously is comparable to that observed with a single immunization as shown in TABLE 2.

EXAMPLE 24 Immunization of mice with AP205 fused C-terminally in GIP peptide 1-15 The DNA fragment encoding the GIP peptide (YAEGTFISDYSIAMD, SEQ ID NO: 27), is formed in three consecutive PCR reactions. In the first reaction, the plasmid pAP405 is used as a template, to amplify the AP205 coat protein gene with oligo pl .45 (5'-AATCTAGAATTTTCTGCGCACCCATCCCGG-3 ', SEQ ID NO: 73) containing an Xba I site, and oligo p4.175 (5'-AATGAACGTGCCCTCTGCGTATCCGGAACCGCCTCCTGC-3 ', SEQ ID NO: 74), to add a nucleotide sequence encoding the amino acid sequence YAEGTFI at the 3' end of the nucleotide sequence encoding GTAGGGSG in the 3 'region of the AP205 coat protein gene in plasmid pAP405. Then, oligos pl.45 and? 4.17ß (5'-CATCGCGATCGAGTAATCGGAAATGAACGTGCCCTCTGCGTA-3 ', SEQ ID NO: 75), are used to amplify the PCR product of the first reaction, thereby adding a nucleotide sequence coding for the amino acid sequence SDYSIAM at the 3 'end of the product of the first reaction. In the third reaction, oligos pl.45 and p4.177 (5'-ACATGCATTAATCCATCGCGATCGAGTAATC-3 ', SEQ ID No. 76) are used to amplify the product of the second reaction thereby adding the 3' end of the second PCR product , a nucleotide sequence coding for the remaining Asp, containing a terminator codon and the Mph 11031 restriction site. The fragment obtained is digested with Xbal and Mphll03I and cloned in the same restriction sites in the vector pAp283 (patent AP205), under the control of the tryptophan operon promoter from E. coli. The resulting structure is: 515 AP205 coat protein - GTAGGGSG - YAEGTFISDYSIAMD The purification method of the expressed fusion protein is substantially the same as that described in EXAMPLE 2 of PCT / EP2005 / 054721. The construction and sequencing of PAP405 and pAP283 is described in PCT / EP2005 / 054721 and in WO2004 / 007538A2, respectively. Adult female mice, C57BL / 6 (5 per group) are vaccinated with AP205 fused C-terminally in peptide GIP 1-15. 50 μg of dialyzed vaccine is diluted in PBS to a volume of 200 μ? and injected subcutaneously (100 μ? on both ventral sides) on days 0, 14, 28 and 42. The vaccine is administered without adjuvant. As a control, a group of mice is injected with PBS or only with AP205 VLP. The mice are exsanguinated retro-biologically on day 0, 14, 28, 42, 56 and 70 and their serum is analyzed by ELISA as described in EXAMPLE 5. TABLE 5 shows the average titers of GIP-specific antibodies. The results shown are the averages of five mice per group. The ELISA titrations are expressed as serum dilutions that lead to the average maximum OD in the ELISA assay. In mice immunized with AP205 fused C-terminally in peptide GIP 1-15, average titers of approximately 1: 184000 were reached by day 42 (TABLE 5). The pre-immunized sera or sera from mice injected with PBS or VLP?) ß showed no sensitization against the GIP peptide. The maximum mean OD titration is less than 100, which was considered to be well below the discriminant value of the assay. This clearly demonstrates that the GIP-VLP fusion is able to induce a high titre of antibodies against GIP fragments, even if it is an autogenous protein. TABLE 5 Average titration of IgG antibodies specific for anti-GIP (expressed as a dilution factor) in mice immunized on day 0, 14, 28 and 42 fused with Ap205 C-terminally to peptide GIP 1-15 Days after the first Immunization Immunization 14 28 42 56 70 AP205 --- GIP 7000 + 900 51000 + 16800 184477 + 59000 156107 + 43000 106100 + 22600 PBS or AP205 100 100 100 100 100 EXAMPLE 25 Effect on glucose tolerance with GIP l-15-GC-0j_ > coupled to VLP?, ß in an animal model of diet-induced obesity To assess the possible effects of glucose tolerance, an oral glucose tolerance test (OGTT) was carried out in vaccinated animals. Briefly, adult C57BL / 6 strand mice (five per group) were vaccinated as described in EXAMPLE 4 with GIP 1-15-GC coupled to the VLP?) ß, which is obtained in EXAMPLE 2. As a control mice were injected with VLP?) ß. A booster immunization was administered on day 122, twenty days before the OGTT. All mice were maintained on a high fat diet for the duration of the experimental period. On day 142 and after 16 hours of the fasting period, the mice were administered a solution of 2 g / kg D-glucose by oral gavage. Mice were bled through the caudal vein at 0, 15, 30, 45, 60, 120 and 180 minutes after oral glucose administration. Blood glucose levels were determined using the Accu-check blood glucose meter (Roche). During this period the mice continued fasting. The blood glucose response kinetics is shown in TABLE 6. No significant difference in glucose response kinetics was observed between the control animals and the control animals. vaccinated. Therefore, vaccination against GIP does not induce a deficient glucose tolerance in mice immunized with GIP 1-15-GC coupled to VLP Qp. TABLE 6 Blood glucose levels in OGTT in 5 mice per group immunized with GIP l-15-GC-Q EXAMPLE 26 Effect on insulin sensitivity with GIP 1-15-60-? ß coupled to VLP? Β in an animal model of diet-induced obesity To evaluate the possible effects of insulin sensitivity, a insulin sensitivity test (IST) in vaccinated animals. Briefly, adult C57BL / 6 strand mice (4-5 per group) are vaccinated as described in Example 4 with GIP 1-15-GC coupled to the VLP?) ß, obtained in EXAMPLE 2. As a control, the mice were injected with VLP? ß. Another booster immunization was administered on day 122, twenty-one days before the IST. All mice were maintained on a high fat diet during the duration of the experimental period. On day 143 and after a 16-hour fasting period, the mice were administered 0.5 U / kg of porcine insulin intraperitoneally (IP). The mice were bled by the caudal vein at 0, 15, 30, 45, 60 and 90 minutes after the administration of insulin. Blood glucose levels were determined using the Accu-check blood glucose meter (Roche). During this period the mice continued fasting. The kinetics of the blood glucose response after IPIST is shown in TABLE 9. Mice vaccinated with GIP 1-15-GC-C $ showed better insulin sensitivity compared to control animals vaccinated with VLP?) ß . The differences were significant at 0, 30, 45 and 60 minutes after the administration of insulin. Therefore, vaccination against GIP improves insulin sensitivity in mice immunized with GIP 1-15-GC coupled to the VLP?) ß. TABLE 7 Blood glucose levels in an IPIST in 4-5 mice per group immunized with GIP 1-15-GC It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (30)

1. CLAIMS Having described the invention as above, the content of the following claims is claimed as property: 1. A composition characterized in that it comprises: (a) A virus-like particle (VLP) with at least one first binding site; and (b) at least one antigen with at least one second binding site, wherein at least one antigen is a GIP protein or GIP fragment and wherein (a) and (b) are linked in at least one first site and at least a second binding site.
2. 2. The composition according to claim 1, characterized in that the GIP protein comprises an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 22; (b) SEQ ID NO: 23; (c) SEQ ID NO: 24; (d) SEQ ID NO: 25; (e) SEQ ID NO: 26; (f) SEQ ID NO: 63; (g) the orthologs corresponding to GIP of any other animals; and (h) an amino acid sequence that is at least 80%, preferably at least 85%, more preferably 90%, or more preferably at least 95% identical with any of (a) to (f)
3. 3. The composition according to claim 1, characterized in that the GIP fragment comprises an amino acid sequence identical to the amino acid residues 7-10 of SEQ ID NO. : 22 (SEQ ID NO: 64).
4. 4. The composition according to claim 1 or 3, characterized in that the GIP fragment comprises an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 27; (b) SEQ ID NO: 29; (c) SEQ ID NO: 32; (d) SEQ ID NO: 45; and (e) an amino acid sequence that is at least 80%, preferably at least 85%, more preferably at least 90% or more preferably at least 95% identical with SEQ ID NOS: 27, 29, 32 or 45.
5. 5. The composition according to claim 1, characterized in that the GIP fragment comprises an amino acid sequence selected from the group consisting of: (a) SEQ ID NO: 28; (b) SEQ ID NO: 31; (c) SEQ ID NO: 43; (d) SEQ ID NO: 44; (e) SEQ ID NO: 68; Y (f) an amino acid sequence that is at least 80%, preferably at least 85%, more preferably at least 90% or more preferably at least 95% identical with SEQ ID NOS: 28, 31, 43, or 44, or 68 6.
6. The composition according to any of the preceding claims, characterized in that the VLP comprises recombinant coat proteins, mutants or fragments thereof or an RNA phage.
7. The composition according to claim 6, characterized in that the phage RNA is a phage RNA?, ß, fr, GA or AP205.
8. The composition according to any of the preceding claims, characterized in that the first binding site is linked to the second binding site by at least one covalent bond, where preferably the covalent bond is a non-peptide bond.
9. 9. The composition according to any of the preceding claims, characterized in that the first binding site comprises an amino group, preferably an amino group of lysine.
10. The composition according to any of the preceding claims, characterized in that the second binding site comprises a sulfhydryl group, preferably a sulfhydryl group of cysteine.
11. 11. Composition in accordance with any of the claims 1-8, characterized in that the GIP protein or GIP fragment is fused to the N- or C-terminus of the coat protein, mutants or fragments thereof, of phage RNA AP205.
12. 12. The composition according to any of the preceding claims, characterized in that it also comprises a linker.
13. 13. A composition characterized in that it comprises: (a) at least one virus-like first particle (VLP) and at least one second virus-like particle (VLP) each with at least one first binding site; and (b) at least one first antigen and at least one second antigen each with at least one second binding site, wherein at least one first antigen is a GIP protein or GIP fragment and at least one second antigen comprises or is a molecule selected from the group comprising: (i) a ghrelin or a ghrelin peptide; (ii) a nicotine, a cotinine or a nornicotine; and (iii) a second GIP protein or a second GIP fragment, wherein the second GIP protein or second GIP fragment is different from the first GIP protein or GIP fragment; and wherein at least a first virus-like particle (VLP) and at least one first antigen are linked through at least one first and at least one second binding site and where at least one second virus-like particle (VLP) and at least one second antigen is linked through at least one primer and at least one second binding site.
14. 14. The composition according to claim 13, characterized in that the first virus-like particle and / or the second virus-like particle comprises recombinant proteins, mutants or fragments thereof, or an RNA phage, wherein preferably the phage RNA is < 2ß, fr, GA or AP205.
15. 15. The composition according to claim 13 or 14, characterized in that the second GIP fragment comprises an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 27 (b) SEQ ID NO: 29 (c) SEQ ID NO: 32 (d) SEQ ID NO: 45 (e) SEQ ID NO: 28 (f) SEQ ID NO: 31 (g) SEQ ID NO: 44 (h) SEQ ID NO: 68 (i) a sequence of amino acids which is at least 80%, preferably at least 85%, more preferably at least 90% or more preferably at least 95% identical with SEAQ ID NOS: 27-29, 31, 32, 44, 68, or 45.
16. 16 The composition according to any of claims 13-14, characterized in that the ghrelin or ghrelin peptide comprises an amino acid sequence selected from the group consisting of: (a) SEQ ID NO: 33; (b) SEQ ID NO: 35; (c) SEQ ID NO: 40; (d) SEQ ID NO: 36; (e) SEQ ID NO: 37; (f) SEQ ID NO: 38; (g) SEQ ID NO: 46 (GSSFLSPEHQKLQ); and (h) SEQ ID NO: 47 (GSSFLSPEHQKVQ).
17. 17. The composition according to any of claims 13-14, characterized in that the second antigen comprises O-succinyl-3 '-hydroxymethyl-nicotine.
18. 18. A kit comprising at least one first composition and at least one second composition, characterized in that it comprises: (a) a first virus-like particle (VLP) with at least one first binding site; (b) at least a first antigen with at least a second binding site, wherein at least a first antigen is a GIP protein or a GIP fragment and wherein (a) and (b) are linked through at least one primer and at least a second binding site; and wherein the second composition comprises: (c) a second virus-like particle (VLP) with at least one first binding site; (d) at least one second antigen with at least one second binding site, wherein at least a second antigen comprises or is a molecule selected from the group consisting of: (i) a ghrelin or ghrelin peptide; (ii) a nicotine, a cotinine or a nornicotine; and (iii) a second GIP protein or a second GIP fragment, wherein the second GIP protein or GIP fragment is different from the first GIP protein or GIP fragment comprised by the first composition; and where (c) and (d) are linked through at least one first and at least one second binding site.
19. 19. The kit according to claim 18, characterized in that the first virus-like particle or the second virus-like particle comprises recombinant proteins, mutants or fragments thereof, of an RNA phage, where preferably the phage RNA is?). , f, GA or AP205.
20. 20. The composition according to claim 18 or 19, characterized in that the GIP fragment comprises an amino acid sequence selected from the group comprising: (a) SEQ ID NO: 27; (b) SEQ ID NO: 29; (c) SEQ ID NO: 32; (d) SEQ ID NO: 45; (e) SEQ ID NO: 28; (f) SEQ ID NO: 31; (g) SEQ ID NO: 44; (h) SEQ ID NO: 68; and (i) an amino acid sequence that is at least 80%, preferably at least 85%, more preferably at least 90%, or more preferably at least 95% identical to SEQ ID NO: 27-29, 31, 32, 44 , 68 or 45.
21. The kit according to any of claims 18-19, characterized in that the ghrelin or ghrelin peptide comprises an amino acid sequence selected from the group comprising: (i) SEQ ID NO: 33; (j) SEQ ID NO: 35; (k) SEQ ID NO: 40; (1) SEQ ID NO: 36; (m) SEQ ID NO: 37; (n) SEQ ID NO: 38; (o) SEQ ID NO: 46 (GSSFLSPEHQKLQ); (P) SEQ ID NO: 47 (GSSFLSPEHQKVQ); and (q) an amino acid sequence that preferably at least 85%, more preferably at least 90%, or more preferably at least 95% identical with SEQ ID NO: 33, 35-38, 40, 46 or 47.
22. 22. The kit according to any of claims 18-19, characterized in that the second antigen comprises O-succinyl-3 '-hydroxymethyl-nicotine.
23. 23. A vaccine composition characterized in that it comprises the composition according to any of claims 1-12 or the composition according to any of claims 13-17.
24. 24. An immunization method characterized in that it comprises administering the vaccine composition of claim 23 to an animal, preferably a dog or a cat, preferably a domestic cat or a human.
25. 25. A pharmaceutical composition characterized in that it comprises: (a) the composition according to any of claims 1-12, the composition of any of claims 13-17, and (b) a pharmaceutically acceptable carrier.
26. 26. A method for producing the composition according to any of claims 1-12, characterized in that it comprises: (a) providing a VLP with at least one first binding site; (b) providing at least one antigen, wherein the antigen is a GIP protein, or a GIP fragment, with at least one second binding site; and (c) linking the VLP with at least one antigen through at least one first binding site and at least one second binding site to produce the composition.
27. 27. The use of the composition according to any of claims 1-12 or the composition of any of claims 13-17 for the manufacture of a medicament for the treatment and / or prevention of obesity.
28. 28. A method of treatment and / or prevention of obesity, characterized in that it comprises administering the vaccine according to claim 23, to an animal, preferably a cat or domestic dog, or a human.
29. 29. A method of treatment and / or prevention of obesity in an animal or a human, characterized in that it comprises administering at least a first composition and at least a second composition of the kit according to any of claims 18-22 to the same animal , preferably a cat or domestic dog, or the same human.
30. 30. The method according to claim 29, characterized in that at least one first composition and at least one second composition are administered at the same time.