JPH09510975A - Synthetic peptide-based immunogen for the treatment of allergies - Google Patents

Abstract

  (57) [Summary] The present invention provides radial branched chain multimeric forms (eg, branched chain octamer or hexamer peptides) or direct chains for inhibiting mast cell activation and reducing allergen-induced IgE production. Use of a composition of synthetic peptide immunogens in the form of a chain monomer in healthy mammals, including humans, of high titer antibodies to effector sites in human IgE heavy chains, ie, in the CH4 domain of the ε chain. A method for inducing production. It also relates to the use of such “optimally” designed carrier protein-free IgEε chain-related immunogens to provide immunotherapy for the treatment of allergies as a key component in synthetic vaccines. The peptide of interest contains an immunostimulatory sequence containing built-in helper T cell epitopes tandemly linked in a specific orientation to help stimulate an immune response to the IgE CH4 domain.

Description

Detailed Description of the Invention                 Synthetic peptide-based immunogen for the treatment of allergies                             Cross-reference of related applications   This was filed January 4, 1991 and is now abandoned with application number 07 / Filed on March 6, 1992, which was a partial continuation application of 637,364, and is currently This is a continuation-in-part application of pending application number 07 / 847,745 that has been abandoned. Pending application number 08 / 060,798 filed May 10, 1993 Pending application No. 08/2 filed March 28, 1994 It is a partial continuation application of No. 18,461.                                 Field of the invention   The present invention relates to the effector site on the CH4 domain of the ε-chain of human IgE heavy chain. For inducing the production of high titer antibodies against it in healthy mammals, including humans A set of synthetic peptides in linear or radial branched multimeric form as immunogens. Wakuchi to provide immunotherapy for use of the product, and treatment of allergies The use of the composition as an agent.                                 Background of the Invention   Igs such as asthma and hay fever known and practiced since the beginning of this century Immunotherapy for the prevention of E-mediated allergic reactions requires increasing amounts of allergen in patients. Desensitization or desensitization to reduce the effects of subsequent exposure to allergens when administered Was due to⁽¹⁾. The limitations of such allergen-based immunotherapy methods are relevant. If the allergen is identified, it can be difficult to identify Side effects often result from the use of controlled allergens⁽²⁾.   Other treatments for relief of allergies are the cellular things that are responsible for the allergic reaction. Use drugs that block the elephant cascade. These drugs include antihistamine , Decongestants, and β₂Agonists, and corticosteroids. Anti Histamine, decongestants, and beta₂Agonists are only in the allergic cascade To It acts on the downstream events of IgE and becomes a palliative therapy that deals only with allergic symptoms. Prevention Therapy must act on cellular events near the onset of IgE-mediated allergic reactions I have to. These palliative treatments provide short-term and partial relief. further Also, relief of symptoms is often accompanied by adverse side effects, for example, antihistamine Results in restlessness or lethargy, β₂Agonists are often It was associated with increased morbidity in breath patients.   Corticosteroids are useful immunosuppressive drugs and are non-useful for the treatment of allergic symptoms. Always valid. However, they stimulate opposite hormonal activity, which is undesirable. It results in widespread immunosuppression.   In order to avoid the disadvantages of known therapeutic agents, selective inhibition by targeting IgE It is more desirable to prevent allergic reactions. This is due to the inconvenient desensitization method. Inhibition of IgE synthesis as is done by, or chemical mediators of hypersensitivity Interrupts the process of stimulating degranulation of mast cells and basophils by the concomitant release of mast cells It is done by either cutting off.   At a more basic level, Stanworth et al.^(3-7)And others^(8-13)Is Groups for immunological triggering of mast cells, mast cells and basophils In an attempt to elucidate the foundation, cell affinity in cell signaling processes To study the role of peptides, synthetic IgE ε-chain peptides and corresponding antibodies Was used.   Of the many IgE peptides (Table 1) studied over the last 20 years, An effective effector site (Lys in the Fc CH4 domain of the human ε chain)₄₉₇-Phe₅₀₆ , Indicated by double underlining in Table 2) was a decapeptide. It was synthesized and used for structure / activity studies⁽³⁾. This IgE CH4 The main-derived tecapeptide is a non-human that mimics IgE-mediated mast cell-induced processes. Dose-dependent hiss from isolated rat peritoneal mast cells in the cell lysis method. Found to have the ability to activate tamin release^(Four). This peptide effect The exact structural requirements for the tractor site require multiple synthetic analogs of the ε-chain decapeptide. Derived through structure-activity studies using^(3,4,5).   Induction by immunization using ε-chain-related "peptide-carrier protein conjugate" The induced anti-IgE CH4 peptide antibody was also observed by observing its inhibitory activity. , Used for structural action studies on degranulation of IgE-sensitized cells^(5,11,12) .   Peptide-based to provide immunotherapy to patients with IgE-mediated susceptibility The possibility of using a vaccine is described by Stanworth et al.^(14,15)By Was suggested. He has keyhole limpet hemocyanin (KLH) or sperm Conjugated with "carrier protein" such as manufactured tuberculin protein fraction (PPD) Lys-Thr-Lys-Gly-Ser-Gly-Phe-Phe-Val-Phe-NH₂ ^{( 3)} Using the previously identified ε-chain decapeptide having the sequence of (SEQ ID NO: 1), Peptide-carrier protein conjugate induced decapeptide-specific antibodies. It turned out. For example, selected based on better than average anti-peptide titer Rabbit anti-peptide serum is a titer-dependent form of decappe from rat peritoneal mast cells. Reduced histamine-induced histamine release. This inhibitory activity is Confirmed by in vivo testing in the anaphylaxis (PCA) model system. The effect of this rabbit anti-peptide serum on anaphylaxis depends on the allergen. Multiple allergen application before administration of anaphylactic antigen ation) to measure blue area when administered to rats presensitized by And by the evaluation of color intensity.   In the same study, such "decapeptide-protein carrier conjugate" The results obtained in rats with immunogens containing A preliminary indication of its potential use as a therapeutic vaccine was given.   However, this strategy encountered considerable difficulties. This Protota The main drawbacks of the ip "decapeptide-protein carrier conjugate" vaccine Is less than optimal and has poorly defined carrier protein and carrier protein Incompatibility due to poor composition and conjugation reaction heterogeneity. You can The antiserum generated by such peptide-protein conjugate is Often more target-peptide, keyhole limpet hemocyanin ( It contains antibodies directed against epitopes on protein carriers such as KLH).   It is known to those skilled in the art that small peptides are poor immunogens. small Chemical conjugation to render various peptides immunogenic , Or by linking the peptide to a large carrier protein by gene fusion . However, these processes generally lead to unpredictable structural changes in peptides. Will occur. Moreover, immune responses are often misinduced to immunodominant carriers. Conclusion As a result, an effective wax that provides long-lasting relief from allergies. The development of Kuching awaits further immunogen design.   In Table 2, rat IgEε chain⁽¹⁶⁾And mouse ε chain⁽¹⁷⁾CH2-CH4 The amino acid sequence for main is the human ε chain⁽¹⁸⁾Along with the amino acid sequence of (SEQ ID NOS: 2-4), for previously reported IgE-related peptide fragments To provide a guide. In the human IgEε heavy chain, the L next to the Q at the 252 position is It is shown to be absent from the zinal IgE myeloma ND sequence. Indicated by a dash Gaps were introduced to maximize homology. Matching homologous residue positions Is surrounded by a square. In Table 2, the positions on the ε sequence studied for structural activity (see Underline (Table 1). Structurally active IgE in the human IgE CH4 domain The CH4 decapeptide sequence is double underlined (SEQ ID NO: 1). A used in the table The mino acid codes are as follows: A, alanine; R, arginine; N, a Sparagin; D, Aspartic acid; C, Cysteine; Q, Glutamine; E, Glu Tamic acid; G, glycine; H, histidine; I, isoleucine; L, leucine; K, lysine; M, methionine; F, phenylalanine; P, proline; S, serine T; threonine; W, tryptophan; Y, tyrosine; V, valine.                             List of references   1. Noon L., "Vaccination against hay fever" (Prophylactic ino culation against hay fever), lancet (Lancet) ， I: 1572-1573 (1911)   2. World Health Organization and International Union of Immunology Working Group Report: Are Current status of Rugen immunotherapy (World Health Organization and International Union o f Immunological Societies Working Group Report: Current status of allerg en immunotherapy.) Lancet, i: 259-261 (1989)   3. Stanworth, Kings, Roy et al., Biochem Cal Journal (Biochem. J.), 180: 665-668 (1979)   4. Stanworth, Molecular Immunology (Mol. Immunol.) ， 21: 1183-1 190 (1984)   5. Stanworth and Burt, Molecular Immunology, 23: 123. 1-1235 (1986)   6. Burt and Stanworth, European Journal of Immunolo Gee (Eur. J. Immunol.), 17: 437-440 (1987)   7. Stanworth, Molecular Immunology, 25: 1213-1215 (1988)   8. Hamburger, Science (Science) ， 189: 389-390 (1975)   9. Kenten, Helm, Ishizaka et al., Procidey Of The National Academy Of Sciences USS I(Proc. Natl. Acad. Sci., USA), 81: 2955-2956 (1984) 10. Coleman, Helm, Stanworth and Gould, Yo -Ropian Journal of Immunology, 15: 966-969 (1985) 11. Chretien, Helm, Marsh et al., Journal of Bu Immunology (J. Immunol.), 141: 3128-3134 (1988) 12. Robertson and Lin, Molecular Immunology , 25: 103-113 (1988) 13. Helm, Kebo, Vercelli et al., Proceedings Of National Academy of Sciences USA, 86: 9 465-9469 (1989) 14． Stanworth, Jones, Lewin and Nyer ayyar), Allergy treatment with a peptide vaccine ptide vaccine), lancet, 336: 1279-1281 (1990) 15. Stanworth, Lewin, Nyer and Jones, Immunoreactive Pep Tides and antibodies and their use in anti-allergic treatment (Immunoreactiv e peptides and antibodies and their use in anti-allergy treatment), EPO 403 312 A1 (1990) 16. Kindsroger et al.DNA 1: 335-343 (1982) 17． Ishida et al.EMBO 1: 1117-1123 (1982) 18. Dorrington and Bennich, immunological ・ Reviews (Immunol. Rev.), 41: 3-25 (1978) 19. Brett et al., European Journal of Immunology, 2 3: 1608-1614 (1993) 20. Weismuller et al., International Journal Of peptide research (Int. J. Peptide Res.) ， 40: 255-260 (1992) 21. Celis et al., Journal of Immunology, 140: 1808-1815 (198 8) 22. Demotz et al., Journal of Immunology, 142: 394-402 (19 89) 23. Chong et al., Infection and Immunity (Infect. Immu n. ), 60: 4640-4647 (1992) 24. Grant, Synthetic Peptides: User's Guide (Synthetic Peptid es: A User's Guide ), W. H. Freeman and Company (W. H. Freeman & Co.), New York, New York, (1992) p. 382. 25. O'Hagan et al., Baxine (Vaccine), 9: 768-771 (1991) 26. Elridge et al., Molecular Immunology, 28: 287-294 (19 91) 27. MacGlashan et al., Journal of Immunology, 136: 2231-2239 (1986)                                 Purpose of the invention   Chemically combined in a radial branched or linear T helper epitope containing form Include humans with a group of IgEε chain-based peptide immunogens made When introduced into a mammal, the decapeptide effector of the CH4 domain of the human ε chain It is an object of the present invention to elicit high titers of antibodies to the -site.   Another object is the introduction of Fg on human IgE CH4 when introduced into mammals, including humans. To a specific orientation that will stimulate the production of high titered antibodies that are effective against Containing a promiscuous human helper T cell epitope Specific derived from human IgE heavy chain CH4 domain (IgE CH4) bound to To design an optimal peptide immunogen having the amino acid sequence of these Antibodies inhibit the activation of mast cells and prevent histamine, which causes allergic symptoms. Which reduces the release of chemical mediators, IgE-mediated passive cutaneous anaphylaxis ( Suppress PCA) reaction and suppress allergen-induced IgE production by B lymphocytes Should be.   A further object is a composition containing a branched multimeric immunogen or a linear immunogen. To study an effective IgEε chain peptide-based vaccine using It is to provide immunotherapy for the treatment of illness.                                 Summary of the invention   According to the present invention, peptide immunogens are prepared by solid phase synthetic methods. The pep Tide immunogen is a 10 amino acid IgE CH4 peptide (SEQ ID NO: 1) or its exemption. A series of radial branched multimeric peptides containing epidemiological analogs; IgE CH4 peptide Peptide (SEQ ID NO: 1) or an immunogenic analog thereof to a helper T cell epitope ( Th epitopes) together with a series of branched multimeric peptides; and Ig Helper T cells with E CH4 peptide (SEQ ID NO: 1) or an immunogenic analog thereof A series of linear monomeric peptides containing together with part of the epitope (Th epitope) It consists of chid. The IgE CH4 peptide is the Fc region of the IgE heavy chain, that is, the ε chain C Obtained from the H4 domain (IgE CH4). Three series of peptide immunogens Of these, linear peptides are preferred. Use compositions containing these peptides And immunize healthy mammals, such as guinea pigs, rats and humans, Is specific for the IgE CH4 effector site (SEQ ID NO: 1) and is absent Induces the production of high titer antisera free of relevant antibodies.   According to the invention, vaccines containing synthetic peptides as key immunogens are suitable Branched multimeric peptides in the presence of various adjuvants and / or delivery vehicles It is also prepared using an effective amount of tide or linear peptide. Such vaccine composition The thing is Stanworth et al.⁽¹⁴⁾Peptides currently used by Induces a more concentrated anti-IgE peptide response than that of single carrier protein conjugates And thus seems to provide a good immunotherapy for the treatment of allergies You.                             Detailed description of the invention   The present invention provides a healthy mammal, including humans, for the treatment of IgE-mediated allergic diseases. Effectors on the CH4 domain (SEQ ID NO: 1) of human IgEε heavy chain in animals Novel group of peptide-based immunogens for the production of high titer antibodies to a single site Regarding the use of   In general, allergic symptoms that are a direct result of IgE-dependent hypersensitivity are Caused by chemical mediators released by cells and basophils. Surface bond Mast cells or basophils sensitized with IgE are specific for surface-bound IgE When bound to the allergen, release is triggered. Induction of allergen, antigen- This is done by the binding of the surface bound IgE to the Fab 'part of the antibody type interaction. Allergen / antibody binding cross-links divalent surface-bound IgE, resulting in high affinity on the cell surface. Of the distal Fc region of IgE, which is the region of IgE in direct contact with the sex Fc receptor Induce a change. Structural changes due to mechanisms that are still not accurately understood Cells with consequent release of histamine-containing mediators by the cells. -Activates the IgE-allergen complex. Effector site on IgE induces It seems to be related to the event. Specific for such "effector site" The presence of selective anti-IgE antibodies is mediated by IgE "antibodies through either active or passive immunization. Alleles by interfering with the interaction between the "fector" site and the cell surface It leads to the inhibition of the cell activation process in a host suffering from a gee reaction.   Such intervention by the use of specific anti-IgE antibodies, a type of immunotherapy, In order to induce the host's own site-specific anti-IgE antibody production by IgE, Passively by prophylactic treatment with specific "site-specific" antibodies against More preferably, a vaccine consisting of a site-specific peptide immunogen is administered to the host. And can be done actively. Active immunization is more effective and long-lasting Seems to provide protection.   Of the sites from the Fc region of circulating IgE studied for functional activity, IgE Region on the CH4 domain of the molecule (Lys₄₉₇-Phe₅₀₆) Is mast cells and halophilic Identified as a structural effector involved in basophil induction^(3-8,14). Table 1 and 2 See the underlined area in the table. Sequence Lys-Thr-Lys-Gly-Ser-Gly-P Derived from this site with he-Phe-Val-Phe-NH2 (SEQ ID NO: 1) The decapeptide was found to be close to the structure of this effector site. this is, Immunological induction process^(Four)From rat mast cells in a manner similar to Proved by the ability of the decapeptide to induce dependent histamine release.   Stanworth et al.^(14,15)Is a carrier protein, -IgE CH4 decapepti linked to whole limpet hemocyanin (KLH) By using an experimental vaccine, The possibility of giving immunotherapy to patients with responsiveness was shown. By such immunization The resulting animal immune sera were obtained from Stanworth et al.^(14,15)By force Decapeptide-induced histamine release from rat peritoneal mast cells in a titer-dependent manner It turned out to reduce it moderately. The inhibitory activity of the obtained immune serum is , In vivo passive cutaneous anaphylaxis (PCA) under conditions of multi-allergen application Therefore, it was confirmed.   The prototype "IgE CH" studied by Stanworth et al. The main drawback of the "4-peptide" vaccine is essentially associated with almost all self-antigens. Its weak immunogenicity, which is a problem.   In the present invention, the synthetic immunostimulatory element has a specific orientation with the IgE CH4 decapeptide. (SEQ ID NO: 1), resulting in a finger at this effector site on IgE. Effective Antibodies Directed to Occur Widely in Genetically Distinct Host Populations Is to provide a specific immunogen capable. These antibodies are then labeled with mast cells. Block the stimulatory effects of IgE on cells and basophils, thus IgE-mediated allergic -Results in effective treatments to prevent the disease.   The peptide immunogen of the present invention comprises a target amino acid sequence of the Fc region of IgE (SEQ ID NO: 1). ) Has the ability to elicit antibodies with serological cross-reactivity with Has the ability to mediate non-cytotoxic histamine release.   Initial dose, eg, 0.2-2.5 mg immunogen; preferably 1 mg, then repeated The (booster) amount should be administered by injection, preferably intramuscular injection You. The dose depends on the age, weight and general health of the patient and will depend on the therapeutic area. Well known.   For active immunization, the term "immunogen" as referred to herein is IgE C Synthetic pep with the ability to elicit antibodies against H4 decapeptide (SEQ ID NO: 1) This antibody is associated with tide and is associated with IgE-mediated basophil and mast cell degranulation. Leads to suppression. The immunogen of the present invention may be a multimeric peptide or a branched chain lysyl coamate. Included its analog with a lix structure.   These branched multimeric peptides independently induce immune responses in host animals. Has the ability to emit. As an analogue of IgE CH4 decapeptide (SEQ ID NO: 1) Is Stanworth et al.^(3.4,5)Synthetic peptides disclosed by Analogues are included (incorporated herein by reference). To be suitable Should have a molecular weight of the immunogen higher than 5,000, preferably 10, Should be higher than 000. The repeating branch unit for peptides is 4 or more Should be.   Bifunctional amino acids such as lysine, followed by preferred non-tiated amino acids such as Gly or Ala. Linkage to amino acids with side chains is useful for creating core matrix structures. is there. One additional coupling between the two di-Boc-lysine coupling cycles. By inserting an amino acid in the Proposal Spacers between peptide branches to allow maximum freedom to obtain the required structure Act as   The immunogen of the present invention contains a promiscuous helper T cell epitope (Th epitope). It also contained a linear peptide with. These Th epitopes are Decapeptide ether with or without spacers to induce The decapeptide N is covalently bound to the effector sequence (SEQ ID NO: 1) in a predetermined form. Adjacent to the end. The immunogen can also be, for example, the bacterium Yersinia sp. (Yersinia spp) -derived immunostimulatory domain of invasin protein It may consist of an array. The invasin domain is a Th epi group via a spacer. It may be attached to the taupe.   The "immunogen" of the present invention minimizes the generation of irrelevant antibodies, thus allergic Without the undesirable side effects that complicate the immunotherapy process for the treatment of , A more focused immune response to the “target sequence”, ie the desired IgE CH4 Induces cross-reactivity (SEQ ID NO: 1). Short peptide antigens are usually T cell dependent It is a sex antigen. That is, the presence of the T helper epitope results in a short "target" peptide. It is needed to make it immunogenic. Short IgE CH4 decapeptide (SEQ ID NO: No. 1) or its immunogenic analogues do not contain T helper cell epitopes. Linear immunity consisting of a branched chain multimeric immunogen and a short IgE CH4 decapeptide Hara was designed to provide artificial built-in functional helper T cell epitopes. Measured.   The peptide immunogen of the present invention has the following formula:             (A)_n-(Th)_m-(B)_o-(IgE CH4 peptide)_p [Where,   A is an amino acid, α-NH₂, Fatty acids, derivatives of fatty acids, or invasion Domain,   B is an amino acid;   Th is a helper T cell epitope or immunopotentiating analog or segment And   The IgE CH4 peptide was Lys-Thr-Lys-Gly-Ser-Gly-Phe-Phe. -Val-Phe (SEQ ID NO: 1) or an immunogenic analogue thereof;   n is 1 to 10;   m is 1 to 4;   o is 0-10;   p is 1 to 3] Indicated by.   The peptide immunogens of the invention are about 20 to about 100 amino acid residues, preferably about 20. 20 to about 50 amino acid residues, more preferably about 20 to about 35 amino acid residues Become.   When A is an amino acid, it may be a non-natural or natural amino acid. Non Natural amino acids include, but are not limited to, β-alanine, ortinine, norleuine. Syn, norvaline, hydroxyproline, thyroxine, γ-aminobutyric acid, homose Examples thereof include phosphorus and citrulline. Natural amino acids include alanine and argi Nin, asparagine, aspartic acid, cysteine, glutamic acid, glutamine , Glycine, histidine, isoleucine, leucine, lysine, methionine, fe Nylalanine, proline, serine, threonine, tryptophan, tyrosine and And valine. Furthermore, n is greater than 1 and two or more A are When it is a noic acid, each amino acid may independently be the same or different.   A is a fatty acid such as stearic acid or palmitic acid or trypa Lumitoyl cysteine (Pam_ThreeWhen it is a fatty acid derivative such as a Cys group, it is , As an adjuvant by enhancing the immunostimulatory properties of Th epitopes Use⁽²⁰⁾. When A is a fatty acid or its derivative, it is usually the amino acid of the peptide. Located at the end. Furthermore, when one of the A's is a fatty acid, the fat useful in the present invention Fatty acids are hydrocarbons of 8 to 24 carbon atoms which may be saturated or unsaturated. Has a chain.   If A is an invasin domain, it is a Yersinia spp. Is an immunostimulatory epitope derived from the invasin protein of. This immunostimulatory property , This invasin domain is a β1 integrin (integrin) present on T cells. ) It results from the ability to interact with a molecule. interacting with β1 integrin The specific sequence for the invasin domain was found to be Brett et al.^{(1 9)} Is disclosed by. In a preferred embodiment, binding to promiscuous Th epitopes. The invasin domain (Inv) for binding has the following sequence: Or to another Yersinia spp. Invasin protein Is an immunostimulatory analog from the corresponding region. Such analogs are substituted , Deletions or insertions may be included to accommodate strain-strain variations. However, the similarity The body retains its immunostimulatory properties.   In one embodiment, n is 4 and A is α-NH₂, Lysine, lysine and And lysine (in that order). In another embodiment, n is 1 and A is α- NH₂It is. In another embodiment, n is 4 and A is α-NH₂, Invasion Domain (Inv), glycine and glycine (in that order).   B consists of the aforementioned natural or unnatural amino acids. Each B is independently the same It may or may not be. When B is lysine, it forms a branched chain polymer be able to. For example, if o is 7 and all 7 B are lysines, Chain K core (K_FourK₂K) is a peptide synthesis without protecting the lysyl side chain ε-amino group. Is formed when. A peptide with a K core has eight branches , Each branch is the same, "(A)_n-(Th)_m− ”Or“ (built-in -IgE CH4 peptide having Th)-". Furthermore, the amino acid of B is , A flexible hinge, or a spacer to form a Th epitope and an IgE C An immune response to the H4 decapeptide or its analogs can be enhanced. flexible Examples of sequences encoding the sex hinge are found in the immunoglobulin heavy chain hinge region. be able to. Flexible hinge sequences are often rich in proline. One A particularly useful flexible hinge has the sequence Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO: No. 24), wherein Xaa is an amino acid, preferably aspartic acid. so is there). Immunogenicity was determined by the promiscuous Th epitope and IgE CH4 decapeptide Improved through the addition of spacer residues (eg, Gly-Gly) between its analogs You can also. The Th epitope is a B cell epitope (ie, IgE CH 4 decapeptide site or its analogs). The lysine residue is the ThE epitope of the IgE CH4 decapeptide (SEQ ID NO: 1). Or it can disrupt the artificial secondary structure created by its binding to its analogues, This can eliminate interferences between T and / or B cell responses. Thus, a structural separation between the helper cell and antibody-inducing domain is provided. Immunogens and more effective interactions between appropriate Th and B cells You.   Th is a Th epitope consisting of natural or unnatural amino acids. Th is It may consist of contiguous or discontinuous epitopes; any amino acid in Th must be It is not always part of the epitope. Including analogues and segments, An epitope suitable for the present invention is the IgE CH4 decapeptide (SEQ ID NO: 1) or It has the ability to enhance or stimulate an immune response against the analog. Immunodominance and And indiscriminate epitopes are widely divergent in animal and human populations MHC type^(21-23)Highly reacts with a wide range. Th suitable for the present invention is about 10 To about 50 amino acids, preferably about 10 to about 30 amino acids. Multiple Th If the epitope is present (ie m>2), each Th epitope is independent And may be the same or different.   Epitope analogs are substitutions of 1 to about 10 amino acid residues in a Th epitope. , Additions, deletions and insertions. The segment comprises the IgE CH4 decapeptide ( Sufficient to enhance or stimulate an immune response to SEQ ID NO: 1) or analogs thereof Is a contiguous portion of the Th epitope.   Epitope of the invention Include hepatitis B surface and core antigen helper T cell epitopes (HB_sTh And HB_cTh), pertussis toxin helper T cell epitope (PT Th), tetanus toxoid Elementary helper T cell epitope (TT Th), measles virus F protein helper T Fine Cell epitope (MV_f Th), Chlamydia trachomati s) Major outer membrane protein helper T cell epitope (CT Th), diphtheria toxin Helper T cell epitope (DT Th), Plasmodium falciparum ( Plasmodium falciparum) circumsporozoite helper T cell epitope (PF Th), Schistosoma man soni) triose phosphate isomerase helper T cell epitope (SM Th), Escherichia coli TraT helper T cell epitope ( TraT Th) and immunopotentiating analogs of any of these Th epitopes and And segments. The following are examples of Th epitope sequences. I can get it: and   Preferably, the Th epitope is HB_sTh, PT₁Th, PT₂Th, TT₁Th, TT_ThreeTh or MV_F1It is Th.   Formula (A)_n-(Th)_m-(B)_o-(IgE CH4 peptide) of the present invention monomer In the linear peptide, the Th epitope is mediated by IgE CH via spacer B. Covalently binds to the N-terminus of the 4 decapeptide (SEQ ID NO: 1). IgE CH4 peptide Is Lys-Thr-Lys-Gly-Ser-Gly-Phe-Phe-Val-Phe (SEQ ID NO: 1), a decapeptide. Immunogenic analog instead of IgE CH4 peptide You may use. The immunogenic analog comprises substitutions, additions, deletions of 1 to about 4 amino acid residues. It may include loss or insertion. However, the analog is the IgE CH4 decapeptide It has the ability to elicit an immune response that is cross-reactive with (SEQ ID NO: 1). Replace, add And insertions are made with natural or unnatural amino acids as defined herein. You may An immunogenic analog of the IgE CH4 peptide (SEQ ID NO: 1) is a stun Worth (Stanworth) et al.^(3,4,5)It is defined by Part of the detailed text.   Therefore, the preferred peptide immunogens of the invention are IgE CH4 decapeptides. (SEQ ID NO: 1) or immunogenic analogue thereof and a monomeric peptide containing Th It is. More specifically, a preferred peptide immunogen is IgE CH4 (SEQ ID NO: 1) or an immunogenic analog thereof; a spacer (eg Gly-Gly); HB_sTh , PT₁Th, PT₂Th, TT₁Th, TT_ThreeTh and MV_F1Th (sequence number No. 5,6,11,7,10,61) Th epitope selected from the group consisting of: And a linear structure optionally containing an Inv domain (SEQ ID NO: 25) is there. Preferred peptide immunogenic compositions include, for example, peptides No. 19-23 and And 28 (Tables 5 and 6, SEQ ID NOs: 51-55, 62).   The peptide immunogens of the present invention are made by chemical synthesis well known to those skilled in the art. May be done. For example, Grant, Synthetic Peptidey Su (Synthetic Peptides)^{(twenty four)}See Therefore, peptide immunogens Ride Biosystems Peptide Synthesizer Model (Applied Biosystems Peptide Synthesizer Model 430A or 431 side chain protection Α-NH protected by t-Boc or F-moc chemistry with protected amino acids₂To The solid-phase synthesis method used is automated using the Merrifield method. May be. To synthesize the K-core moiety that synthesizes the peptide branch, t-Boc lysine Di-α, ε (t-Boc) lysine residue instead of 2,4-dichlorobenzyl protected ε- Used with amino acids.   When A is a fatty acid, it is resin-bonded by the well-known carbodiimide method. It is easily added to the N-terminus of peptides. Pam_ThreeTo add Cys, lipoami Sano acid S- [2,3-bis (palmitoyloxy)-(2R) -propyl-N-palmito Il- (R) -cysteine (Pam_ThreeCys) is chemically synthesized. Then Pam_ThreeCys In the final coupling step_ThreePeptides by solid-phase synthesis method using Cys-OH Coupling to the N-terminal of tide to attach lipoamino acid to resin-bound peptide chain Let it.   To improve the solubility of the last bound lipopeptide product, solid phase peptide Can be extended at the N-terminus with additional serine and lysine residues.   After completing the assembly of the desired peptide immunogen, treat the resin using standard methods. Release the peptide from the resin and deprotect the functional group on the amino acid chain. Free The peptide is purified by HPLC and biochemically, eg by amino acid analysis or Characterize by sequencing method. Purification and characterization methods for peptides Are well known to those skilled in the art.   Other chemistries for obtaining the linear Th-IgE CH4 decapeptide constructs of the present invention Haloacetylated via formation of a "thioether" linking group Examples include ligation of cysteinyl peptides. For example, cysteine is It can be added to the C-terminus of Th-containing peptides and uses the thiol group of cysteine. And IgE CH4 decapeptide (SEQ ID NO: 1) or its immunogenic analog N N of lysine residue bound to the end^αChloroacetyl modification or maleimide-derived α- Or ε-NH₂A covalent bond can be formed to an electrophilic group such as a group.   The peptides of the invention can also be polymerized. Polymerization is carried out, for example, by a conventional method. Using -NH of glutaraldehyde and lysine residue₂By the reaction between groups be able to. Linear "A-Th-spacer-IgE CH4" peptide construct (For example, Peptide Nos. 19-23 and 28, SEQ ID NOs: 51-55 and 6 2) is added to the N-terminus of the linear "A-Th-spacer-IgE CH4" construct. It may be polymerized or copolymerized by the use of the further cysteines mentioned. No. The thiol group of the 19-23 terminal cysteine has a branched poly-lysyl core molecule (eg, For example, K₂K, K_FourK₂K, K₈K_FourK₂H) of lysine residue bound to the N-terminus of K) Loroacetyl modification or maleimide-derived α- or ε-NH₂With group "Thioe Used for the formation of the "Toll" bond.   Alternatively, the well known recombinant DNA method may be used for long linear peptides. The immunogen may be synthesized. The standard manual for the DNA method is the present invention. Provides a detailed protocol for producing the peptides of The peptide of the present invention In order to construct a gene encoding Reverse amino acid sequence to nucleic acid sequence using organism-optimized codon usage translate. It then typically encodes peptides and modulators, if desired. A synthetic gene is created by synthesizing overlapping oligonucleotides. The sum Insert the gene into a suitable cloning vector to obtain recombinants for characterization. U. The peptide is then expressed under suitable conditions appropriate to the chosen expression system and host. Let it manifest. The peptide is purified and characterized by standard methods.   The potency of the peptide immunogens of the invention was demonstrated by injecting the immunogens into animals as shown in the examples. And then IgE CH4 decapeptide (SEQ ID NO: 1) or immunogenic analogues thereof It may be established by monitoring the humoral immune response to the body. Suitable Animals include mice, rats, rabbits, guinea pigs, pigs, goats, sheep, etc. Is mentioned.   Another aspect of the invention is the use of an effective amount of a peptide of the invention in a pharmaceutically acceptable delivery system. It is intended to provide a vaccine composition comprising one or more types of peptide immunogens. Scarecrow Vaccine composition for preventing atopic reaction including allergic rhinitis, food allele Other IgE media such as ghee, asthma, anaphylaxis, and virus-induced asthma Used for prevention of hypersensitivity reaction.   Thus, the peptide immunogens of the invention are adjuvants, pharmaceutically acceptable Using carrier or other ingredients conventionally provided in vaccine compositions, It can be formulated as a tin composition. Such formulations are easily prepared by those of ordinary skill in the art. The formulation is for immediate release and / or sustained release, and Formulations for induction of systemic immunity and / or induction of local mucosal immunity include This is done, for example, by microparticle immunogen entrapment. You. The preparations include alum, Freund's incomplete adjuvant, liposin, saponi , Squalene, L121, Emulsigen and ISA 720 etc. Agents or emulsifiers may be included.   Vaccines of the invention can be administered subcutaneously, orally, intramuscularly, or by other parenteral or enteral routes. Is administered by any convenient route including. Administered as a single dose or multiple dose Is done. The immunization schedule is easily determined by one of ordinary skill in the art.   The vaccine composition of the present invention comprises an IgE CH4 decapeptide or an immunogenic class thereof. Effective amount and pharmaceutically acceptable amount of one or more synthetic peptide immunogens, including analogues Contains a carrier. The dosage unit form is about 0.5 μg to about 1 mg of each peptide per kg of body weight. To be formulated. When delivered in a multidose drug, The effective dose is conveniently divided so as to contain the appropriate amount per unit dosage form. .   The vaccine composition of the present invention enhances immune efficacy in the more general population, thus In order to provide a good immunogen against IgE CH4 decapeptide It may be formulated so as to contain a cocktail of the above-mentioned subject peptide immunogens. For example, a cocktail of peptides No. 19, 20, 21, 23 and 4 is useful You. The composition comprises a lipopeptide to provide a built-in adjuvant. May be formulated as. Against immunogens containing synthetic IgE CH4 decapeptide Immune reaction is based on O'Hagan et al.^{(twenty five)}Raw of the type disclosed by Delivery via entrapment in or on biodegradable microparticles May be improved by The immunogen is covalently bound to Pam._ThreeAju including Cys Encapsulation may be with or without bunting and such fine particles may be Incomplete adjuvant or immunostimulatory adjuvant such as alum Good. The microparticles are effective in the immune response to immunogens, including local mucosal immunity. Function to increase. Such local immunity can be defined, for example, by mucosal topical allergic -It is particularly desirable because of the reaction. The vaccine composition in microparticulate form should be persistent or Orally and locally to give a time-controlled release for cyclical reactions. May be formulated for local administration^(25-26).   The present invention will be described below with reference to specific peptide immunogens. It is not used to limit the range.                                 Example 1                        Synthesis of octameric peptide immunogen   The following multimeric peptides were synthesized:   Peptide No. 1   Peptide No. 2   The synthesis of multimeric peptides involves the synthesis of low molecular weight matrix cores with branched multimeric peptides. A limited sequence of trifunctional amino acids to supply as they are the basis for the formation of the antigen system. It proceeds by sequential propagation. Trifunctional amino acid, sun That is, Boc-Lys (Boc) or di- (Boc) -Lys is N^α-And N^ε-Ami Most suitable because both acid groups are available as reactive ends. Thus , Continuous di- (Boc) -Lys extension is 2ⁿWill generate one reactive end.   For example, the initial coupling of di- (Boc) -Lys onto a solid phase resin gives two Creates a reactive amino terminus of 2 to join the two peptide chains. Therefore, By the continuous production of the second, third and fourth steps using (Boc) -Lys, a large amount of The four-, eight- and sixteen-valent ends for binding the somatic peptide chain antigen are generated. Would. Such multimeric peptides are useful as immunogens. Use as an immunogen For this purpose, the branched octamer peptides No. 1 and 2 were synthesized. The branched chain The antigen is surrounded by a dense layer of peptide-antigen that is homogeneous around the core matrix. It contains a small heptalysyl core entrapped. This design has many uncertainties. On a large protein carrier such as PPD or KLH and the surface of the protein carrier Conventional peptide-carrier conjugates containing randomly distributed small peptide antigens Digested antigen is different.   Synthesis of octameric peptide immunogens was performed using Boc-amino acid resin-bound benzhydrylamine. And a combination of tBoc-chemistry is used. For example, Biosearch 9500 equipment with a very low loading of 0.14 mmol / g MBHA (4-methylbenz (8) by coupling di-t-Boc Lys to a hydrylamine) resin. -A branched heptalysyl core resin was prepared. For each of the di- (Boc) -Lys Two coupling cycles followed by 0.3M in DMF dimethylformamide Two capping reactions with acetylimidazole were performed.   First di- (NH₂) Lys-resin to two more di- (Boc) -Lys couplings Was added. Then the substitution level of the synthetic octamer resin was determined by the ninhydrin test. Measured and appropriate level of free-NH based on theoretical coupling yield₂Have a group And then the octamer peptide immunization according to the standard t-Boc method. Used for original synthesis.   Acid-labile tert-butyloxycarbonyl (for protection of N-α amino acids ( t-Boc) was used. For the following functional side chain protecting groups: Thr, Ser, Glu and Tyr. For O-benzyl; N for Arg^α-Tosyl; BOM for His, That is, BOC-N^im-Benzyloxymethyl, N for Lys^ε-Zikuro Robenzyloxycarbonyl; for Cys, S-4-methylbenzyl-; Asp O-cyclohexyl for and CHO for Trp.   Directed by the sequences of peptides No. 1 and No. 2 (SEQ ID NOs: 23, 13) Thus, from the C-terminal to the N-terminal, consecutive amino acids of peptides No. 1 and No. Nono acid was added. Octamers obtained for peptide No. 1 and peptide No. 2 Peptidyl resin was treated with anhydrous HF in the presence of 10% v / v anisole at 0 ° C. for 1 hour. Cut by. The released multimeric antigen is extracted with acetic acid and washed twice with ether. And lyophilized. Lyophilized multimeric peptite was used as the immunogen.                                 Example 2   Branched octamer peptide immunization with CFA and IFA as adjuvants Active immunization by Hara   (A) Immunization method   Groups of guinea pigs (N = 3 per group) according to the following protocol ) Were labeled with two IgE CH4 related multimeric peptide immunogens (peptide No. 1 and 2), as well as peptide No. 3 (SEQ ID NO: 1) conjugated to KLH. ) With a peptide-based immunogen or conjugate (100μ) g / mL) emulsified with the same amount of Freund's complete adjuvant (CFA) as a mixture (2 00 μL) was injected subcutaneously into each animal. Then on days 21, 42 and 63 Peptide-based immunity mixed with incomplete Freund's adjuvant (IFA) in the eye Repeated subcutaneous injections of stock were made.   (B) Guinea pig immunization by measuring anti-IgE CH4 related peptide response Serum assay   Using 96 well flat bottom microtiter plate coated with corresponding immunogen And measure the anti-peptide antibody activity by ELISA (enzyme-linked immunosorbent assay) I do. Aliquat (100 μL) of 5 μg / mL peptide immunogen solution was added to 37 Incubated at 0 ° C for 1 hour. The plate was mixed with 3% gelatin / PBS solution. Blocked by further incubation for 1 hour at 37 ° C. Next The blocked plate was dried at and used for the assay. Test guinea pig Aliquots of serum (100 μL) were diluted 1:10 in sample dilution buffer. Therefore, 10-fold serial dilutions were then added to the peptide-coated plates. The plate Were incubated at 37 ° C for 1 hour. Use normal guinea pig serum as a control Was. Was. Horseradish peroxidase-labeled goat anti-guinea pig antibody 100 μL Containing conjugate dilution buffer (0.5M NaCl and normal goat serum) Phosphate buffer) at a dilution of 1: 1,000. Plate at 37 ° C for 1 hour After incubation for a period of time, it was washed as described above. Then o-phenylenedia Aliquote (100 μL) of Min substrate solution was added. Color development for 5 to 15 minutes 2NH₂SO_FourThe enzymatic coloring reaction was stopped by the addition of 50 μL. A of the contents of each well with a plate measuring instrument_492nmWas measured.   Peptide No. 1 and its highly related, both branched chain multimeric immunogens Derivative peptide No. 2 was treated with IgE CH4 through an ELISA inhibition assay. Be effective in inducing antibodies specific for the target sequence (SEQ ID NO: 1) found. Monomer peptide No. 3 (IgE CH4 decapeptide) as a control immunogen When compared to the KLH conjugate of SEQ ID NO: 1), the results show that these two Multimeric peptide antigen produced higher levels of antibody titer than KLH conjugate That was shown.   The successful result of these immunization experiments was the Gly-Lys-Met at the C-terminus of the peptide sequence. (See SEQ ID NOS: 23 and 13, peptide Nos. 1 and 2) And showed the occurrence of Th epitopes in the multimeric system, and provided effective provision to the immune system. It showed the importance of tedious orientation. In other experiments, simply in other orientations IgE 8-or containing a CH4 decapeptide (SEQ ID NO: 1) or multiple repeats thereof Only produces the 16-branched IgE peptide immunogen, It was shown that it was not effective in inducing response. In fact, a total of 19 branched chains Of the body constructs, peptides No. 1 and 2 are the only ones showing enhanced immunogenicity Things. In this regard, it was observed with the multimeric peptides No. 1 and 2. High immunogenicity requires experimentation and was not expected by one of ordinary skill in the art.   In addition, the results obtained show that the spacers incorporated between the short IgE CH4 segments. Sequence, that is, Gly-Pro-Gly, is provided by the subunit sequence. It is shown that the free provision of the selected epitope is required. Branched chain lysine core resin Insertion of a spacer at the C-terminus, namely Gly-Lys-Met, before ligation to Immunogenicity of the synthetic branched chain IgE CH4 decapeptide (SEQ ID NO: 1) synthetic construct It also turned out to be necessary.                                 Example 3          Immunization of rats with a linear immunogen (SEQ ID NOs: 15-22)   A.Preparation of immunogen: Applied Biosystems Peptide Synthesizer Iser Model (Applied Biosystems Peptide Synthesizer Model) 43 Solid phase synthesis using F-moc chemistry according to manufacturer's instructions at 0A or 431. To synthesize peptide immunogens A to H (Table 3). Complete assembly of the peptide The resin is then treated according to standard methods to cleave the peptide from the resin, Deprotects the functional group on the amino acid side chain. From the amino terminus to the carboxyl terminus The structure of the peptide immunogen is as follows: Peptide Immunogen A is Main: 3 lysine residues (3K), hepatitis B surface antigen helper T cell epitope (HB_sTh epitope) and a linear peptide having an IgE CH4 peptide It is. Thus, peptide immunogen A is 3K-HB_sTh-IgE CH4 pepti Indicated as Do. For peptide immunogen A and for peptide immunogens BH The actual sequences are shown in Table 5 (SEQ ID NOs: 15-22).   For immunizations at 0, 2, and 5 weeks, each peptide immunogen was lysed and Tubant solution (Freund's complete adjuvant, Freund's incomplete adjuvant, Or 0.2% alum) to give a final concentration of 0.5 mg / ml. Use the solution Store at 4 ° C until use and vortex 3-5 minutes prior to injection. Injection into each rat Or 100 μg is administered.   B.Immunization schedule and serum collection:   Sprague Dawley rats (n = 5) are immunized subcutaneously (sc). Booster injections are given sc at 2 and 5 weeks. 3rd week, 6th week, Blood is collected at 7 and 11 weeks.   Sodium pentobarbital (64.8) diluted 1:10 in 0.9% NaCl. mg / mL; Anthony Products Co., Ltd. After anesthetizing the rat by intraperitoneal injection of 1 mL, Akkadia, Li. Blood is collected from the middle caudal artery. The tail is 48 ° C ± 0. Keep in water at 5 ° C for 2 minutes and massage quickly with a paper towel (ie Exploit). Immediately collect blood in a 5 mL syringe fitted with a 23 gauge needle. You. Typically, 2-2.5 mL blood is obtained. Centrifuge for 25 minutes at 3000 rpm The serum is collected. Aliquot the serum into 300 μL volumes and Store frozen until used for LISA assay.                                 Example 4          Immunization of rats with a linear immunogen (SEQ ID NOs: 37-50)   A-Th-GG-IgE CH4 [where A is NH₂-, Lys-Lys (2K), Lys-Lys-Lys (3K) or invasin domain (Inv) (SEQ ID NO: 25 ), Th is a T helper peptide, and GG is Gly. -Gly spacer, IgE CH4 is a target decapeptide (SEQ ID NO: 1) The linear peptide immunogens shown in Figure 3 are synthesized as described in Example 3. This These peptide immunogens are shown in Table 4 as peptide immunogens No. 4-17 (SEQ ID NO: 37-50). Immunize rats with synthesized and cleaved peptides And test for efficacy.   By the experimental immunization protocol outlined below, 5 rat groups were tested. Efficacy is evaluated for the group.   Experimental design:             Immunogen: Peptide No. 4-17 (1 per trial)             Dosage: 100 μg per immunization               Pathway: intramuscular       Adjuvant: Freund's complete / incomplete           Dosing schedule: Week 0 (FCA), Week 3 and 6 (IFA)           Collection plan: Weeks 0, 3, 6, 8, 10                 Species: Sprague Dawley rat     Group size: 5           Assay: ELISA for anti-peptide activity, solid phase immunosorbent                     , The monomer of the IgE CH4 decapeptide sequence (SEQ ID NO: 1)                     Peptide No. 3.   Blood was collected, processed into serum and stored, then goat anti-guinea pig as a tracer. Horseradish peroxidase labeled goat anti-rat IgG instead of IgG Titer by ELISA as described in Example 2 except that antibody is used.                                 Example 5   Linear immunogens (SEQ ID NOs: 51-56, 62) and reverse polarity linear immunogens (SEQ ID NOs: 51-56, 62). Immunization of rats according to columns No. 57-60)   Peptide immunogens No. 18-23 (Sequence shown in Table 5 as described in Example 3 No. 51-56) was synthesized. The formula for the peptide immunogen No. 18-23 is: A-Th-GG-IgE CH4 [wherein A is N-terminal, Lys-Lys (2K), Lys- Lys-Lys (3K), or an spacer separated from the Th sequence by a spacer GG. -Thin domain (Inv) (SEQ ID NO: 25); Th is HB_sTh, PT₁Th, PT₂Th, MV_F1Th or TT_ThreeSelected from the group consisting of Th; GG is Gly -Gly spacer; IgECH4 is an IgE CH4 decapeptide (SEQ ID NO: No. 1)]].   In the same manner as peptide No. 18-23, peptide No. 24- in Table 5 was used. A peptide immunogen of SEQ ID NOs: 57-60, also shown as 27, was synthesized. these Peptide is designated as IgECH4-GG-Th. These peptides are Peptide / Th polarity reversed, ie, IgE CH4 decapeptide (SEQ ID NO: No. 1) is at the N-terminus and Th is at the C-terminus, except for the IgE CH4 decape Regarding peptides, peptides Nos. 19, 20, 21, 23 (Table 5), spacers and And Th array.   Using these peptide immunogens, for comparison and demonstration of efficacy, Rats were immunized as described in the experimental protocol.   Experimental design:             Immunogen: Peptide No. 18-28 (1 per croup)                     (SEQ ID NOS: 51-60 and 62)             Dosage: 100 μg per immunization               Pathway: intramuscular       Adjuvant: Freund's complete / incomplete for peptide No. 18-27,                     0.4% alum for peptide No. 28           Dosing schedule: Week 0 (FCA) 3 and 3 for peptide No. 8-27.                     And 6th week (IFA), 0 and 3 for peptide No. 28                     And 6th week alum           Collection plan: 0, 3, 6, 8, 10 weeks                 Species: Sprague Dawley rat     Group size: 5 for peptide No.27-28, to peptide No.28                     About 4           Assay: ELISA for anti-peptide activity, solid phase substrate                     Cide No. 3 (SEQ ID NO: 1) Blood collected, processed into serum and stored, then goat anti-guinea pig as tracer Horseradish peroxidase labeled goat anti-rat IgG anti-IgG instead of IgG The titer was determined by ELISA as described in Example 2 except that the body was used. all All sera were assayed by anti-peptide ELISA and diluted 1: 100>0. A of 2_492nmThose samples that gave a value were recorded as seropositive.   Immune efficacy of peptide immunogen No. 18-28 (SEQ ID NOs: 51-60 and 62) Were evaluated by anti-peptide ELISA and were 6 and 8 weeks depending on experimental immunization. Seropositive reactivity for IgE CH4 peptide No. 3 by eye (ie, 1:10 A at 0 dilution _492nm >Of rats in each group of 4 or 5 converted to 0.2) The numbers are shown in Table 6.   The peptide immunogen of this example of polar Th-GG-IgECH4 (Peptide No. 1 8-23 and 28, SEQ ID NOs: 51-56 and 62) are IgE CH4 decappe. Significant potency for the induction of antibodies against peptide (peptide No. 3, SEQ ID NO: 1) showed that. Immunize with this polar peptide immunogen (Peptide No. 18-23, 28) All 6 groups of rats showed significant conversion to seropositivity compared to controls did. Seroconversion prevalence for the group was reduced from 1/5 to 5/5 by 6 weeks The seroconversion morbidity varies at weeks 6 and 8 depending on the third dose of immunogen. In the meantime it continued to increase. HB_sA peptide immunogen No. 18, containing a Th peptide sequence, MV_F1Peptide immunogens No. 19 and PT with Th peptide₁Th peptide distribution Peptide No. 28 containing rows was 4/5, 5/5 and by week 8, respectively. It was the most effective with a seroconversion rate of 4/4. Peptide immunogen No. A comparison of the immunogenicity of 21 and 22 (SEQ ID NOs: 54, 55) is shown by the Inv domain peptide. Cido PT until the 8th week₂Provides a significant improvement in the immunostimulatory capacity of Th-containing peptides This is shown (Table 6).   In contrast, a similar peptide immunogen with reverse Th polarity (Peptide Immunogen No. 2 4-27, SEQ ID NOs: 57-60) have significant immunopotency on seroconversion in rats. Is not shown. This low immunopotency was derived from Th-GG-IgECH4 amino. The polarity to the syl terminus is crucial for the immunogenicity of the linear peptide immunogens of the invention. Indicates that there is. Target peptide and Th in one direction over another The determination of efficacy was unexpected and unexpected for those skilled in the art.                                 Example 6           Linear immunogen cocktails further expand responsive population   Many different linear constructs containing IgE CH4 decapeptide and Th ( Cocktail vaccine compositions by establishing the relative potency of Examples 3-5) Allows the selection of useful peptide immunogens for formulating Measles virus Immunodominant promiscuous derived from F protein, tetanus toxin and pertussis toxin ) Individual Th-GG-IgECH4 constructs with Th peptite (Peptite No. 1 9-23) was added to the IgECH4 decapeptide (SEQ ID NO: It was proved to be effective in inducing an antibody response to 1). these A composition containing a mixture of linear peptides of It may provide the desired maximum immunogenicity.   Preferred "A-Th-GG-IgECH4" polar synthetic peptides, pepti Immunogen No. 19, 20, 21, 23 (Table 5) and peptide immunogen No. 4 ( Immunization of such compositions formulated to contain the mixture of Table 4, Example 4) Efficacy was evaluated in rats by the protocol described in Example 5. Luck Each animal in a group of 5 animals was treated with 10 equimolar formulations of 5 peptides. Immunizations were performed with a dose of 0 μg, ie 20 μg of each peptide. 5th and 8th week The number of rats converted to seropositive reactivity by 5 was 5 out of 5 (at both time intervals). That is, it was 100%).   The results show that the vaccine comprising the cocktail of peptide immunogens of the present invention was improved It is shown to provide immunogenicity. Such as a cocktail of individual active peptides The potency for this mixture has been shown to be immunotherapeutic in a genetically diverse human population. It is also shown to induce a body reaction.                                 Example 7                 Immunization with a cocktail of effective linear immunogens   Many different linear constructs containing IgE CH4 decapeptide and Th ( By establishing the relative potency of Examples 3-5), for the cocktail of immunogens It is possible to select a peptide useful for. Gly-Gly Spec for Immunogen Cocktail And measles virus F protein, hepatitis B surface antigen, tetanus toxin and Individual constructs with promiscuous Th peptides derived from pertussis toxin should be effective Is shown (Table 6). These straight chains with specific polarities with proven potency A mixture of peptide-like immunogens is thus maximally immune to genetically diverse populations. Can provide epidemiology. The following protocol describes the preferred "A-Th-GG- Formulated as a mixture of synthetic peptide immunogens containing the "IgECH4" construct It was designed to show efficacy for the compositions of the present invention.   Experimental design:             Immunogen: (1) Cocktail 1: Peptide No. 18, 19, 20                     (2) Cocktail 2: Peptito No. 18, 19, 22                     (3) Positive control-peptide 3 KLH conjugate                           (1 immunogen per group of rats)             Dose: Equivalent to 100 μg or 33.3 μg peptide per immunization                     Molar equivalents of each synthetic peptide or IgE CH4 equivalent of               Pathway: intramuscular       Adjuvant: (1) Freund's complete / incomplete                     (2) 0.4% alum (aluminum hydroxide)                     (Any one adjuvan per immunogen per group                     G)           Dosing schedule: Week 0, 2 and 4                     (The CFA / IFA group received CFA at week 0                     IFA is administered at weeks 2 and 4. Alum                     The alum preparation is administered to all three doses)           Collection plan: Weeks 0, 3, 6 and 8                 Species: Sprague Dawley rat / group     Group size: 5, 6 groups           Assay: ELISA for anti-peptide activity, solid phase immunosorbent                     , Peptide No. 3 (SEQ ID NO: 1).   Blood was collected, treated with serum and stored before ELI as described in Example 5. Titered by SA.   This experiment^(14,15)Of the immunogens of the invention compared to known immunogens of Planned to show good performance. The results include three Th peptides each. Useful for the evaluation of two mixtures of effective peptide immunogens having Utility of the fragile Inv domain (Cocktail 2 contains Inv, Cocktail 1 contains Not) and alum as an adjuvant in the vaccine composition of the present invention Shows the efficacy of.                                 Example 8                     Clinical trials using immunogen cocktails   Many different linear constructs containing IgE CH4 decapeptide and Th ( By establishing the relative potency of Examples 3-5), for the cocktail of immunogens It is possible to select a representative peptide of Gly-Gly in immunogen cocktail Pacer and measles virus F, hepatitis B surface antigen, tetanus toxin and pertussis The individual constructs containing the toxin-derived Th peptide sequence showed efficacy ( Table 6), indiscriminately for multiple human HLA DR antigens, resulting in genetic To provide maximum immunogenicity in a diverse human population. Furthermore, these T Since h-peptide is derived from a child's vaccine, vaccination at an early age It is an effective source of Th memory in the immunized human population. Thus, the child's work Kutin against an anti-allergic vaccine consisting of a mixture of such Th peptides Has potency to provide enhanced immune efficacy. The clinical protocol below is Such a linear "A-Th" in alum, a widely accepted adjuvant -Spacer-IgECH4 decapeptide "formulated as a mixture of peptide immunogens It was designed to show efficacy for formulated compositions of the present invention.   Experimental design:               Target: Patients with hay fever     Season and period: Hay fever season, 8 weeks           Group: 4 groups, 1 group / immunogen / administration                     N = 15, 12 per group received immunogen and 3                     Administer placebo.             Immunogen: Cocktail 1: Peptide No. 18, 19, 20, 23       Adjuvant: 0.2% alum             Dose: each equivalent to 500 μg or 125 μg peptide per dose                     Synthetic peptide molar equivalent               Pathway: intramuscular           Dosing schedule: Week 0, and Week 4           Evaluation plan: Weeks 0, 4 and 6 Blood was collected, treated with serum, stored and then subjected to ELIS as described in Example 5. Titered by A.   For skin reaction tests, the rate of patient use of hay fever medication, allergic symptoms and side effects Physical examination of the patient as well as the subjectivity of the effect of using the product in an interview with the patient. And safety of vaccine composition "Cocktail 1" by obtaining positive evaluation Sex is assessed serologically. For serological evaluation, the previous ELISA, and to measure reduction in histamine levels and the product Standard automated spectrofluorimetry to confirm that thyroid does not induce histamine release Routine assays. Skin tests consist of injecting a standard solution of allergen into the upper layers of the skin. It is a percutaneous test for irradiation. A typical "generated in the reaction to an allergen The response to the allergen was determined by measuring the area of wheals and flushes. Quantify. Expected results include a significant improvement in allergic symptoms at the end of the study , Without evidence of histamine release induced by the vaccine composition of the present invention .   This experiment is designed to demonstrate the clinical efficacy of the present invention. The result is medically acceptable 4 Th peptide sequences formulated with alum, an included adjuvant Of "A-Th-spacer-IgE CH4 decapeptide" immunogen containing Provides a rating of things.                                 Example 9   In vitro assay shows potency of IgE CH4 decapeptide specific antibody   Antibody inv elicited by immunization with IgE CH4 decapeptide immunogen Passive sensitization in the well-known histamine release assay for itro assessment Human basophils are used. Preparation of human basophils from venous blood of volunteers , And then hyperimmunoglobulin blood against BPO-HSA specific IgE (Hyperimmunoglobulemic) benzylpenicilloyl-prepared from donor blood- IgE specific for human serum albumin conjugate (BPO-HSA) Sensitize yourself with. Histamine release by sensitized basophils was found in BPO-HSA Alternatively, it is performed by administration of IgE CH4 peptide No. 3 (SEQ ID NO: 1). Hi Basophils were treated with IgE CH4 decapep before addition of drugs to induce stamin release. Combine with serial dilutions of antiserum or preimmune control serum against Tide (SEQ ID NO: 1) . Samples are analyzed for histamine release by automated fluorescence. Spontaneous release Histamine release from the percentage of total basophil histamine content of the sample after subtraction from both Calculate the percentage of⁽²⁷⁾. The ability of experimental antisera to inhibit histamine release is  In vitro potency performance.   This assay demonstrates that the concentration of IgE CH4 peptide No. 3 (sequence The histamine release-inducing ability of No. 1) was shown. The results shown in Table 7 are IgE C H4 peptide No. 3 (SEQ ID NO: 1) induces human release by human basophils and To confirm the relationship of antibody to column number 1 and the corresponding human allergic reaction. It was offered for the purpose.

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Claims (1)

[Claims]   1. formula:             (A)_n-(Th)_m-(B)_o-(IgE CH4 peptide)_p [Where,   A is an amino acid, α-NH₂, Fatty acids or their derivatives, or invasin Is;   B is an amino acid;   Th is a helper T cell epitope, analog or segment thereof;   The IgE CH4 peptide is SEQ ID NO: 1 or an immunogenic analog thereof;   n is 1 to 10;   m is 1 to 4;   o is 0-10;   p is 1 to 3] The peptide immunogen represented by.   2. The peptide immunogen according to claim 1, wherein p is 1.   3. Th is SEQ ID NOs: 5-12, 14, 26-36, 61 and immunogens thereof The peptide immunity according to claim 1, which is selected from the group consisting of sex analogues or segments. Epidemics.   4. Th is SEQ ID NOs: 5-12, 14, 26-36, 61 and immunogens thereof A peptide immunity according to claim 2 selected from the group consisting of sex analogues or segments. Epidemics.   5. Consisting of SEQ ID NOs: 13, 15-23, 37-50, 51-56 and 62 The peptide immunogen according to claim 1, which is selected from the group.   6. The method of claim 3 selected from the group consisting of SEQ ID NOs: 51-56 and 62. Peptide immunogen.   7. The peptide immunogen according to claim 1, wherein A is a fatty acid.   8. The peptide immunogen according to claim 2, wherein A is a fatty acid.   9. The peptide immunogen according to claim 1, wherein A is a fatty acid derivative.   10. The peptide immunogen according to claim 2, wherein A is a fatty acid derivative.   11. Fatty acid derivative is Pam_ThreeThe peptide immunogen according to claim 9, which is Cys.   12. Fatty acid derivative is Pam_ThreeThe peptide immunogen according to claim 10, which is Cys.   13. An effective amount of the peptide of claim 1 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   14． An effective amount of the peptide according to claim 2 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   15. An effective amount of the peptide of claim 3 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   16. An effective amount of the peptide according to claim 4 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   17． An effective amount of the peptide according to claim 5 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   18. 7. A peptide according to claim 6 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   19. An effective amount of the peptide according to claim 7 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   20. A peptide according to claim 8 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   21. An effective amount of the peptide according to claim 9 in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   22. 11. A pepti according to claim 10 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   23. The pepti according to claim 11 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   24. 13. A pepti according to claim 12 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising an immunogen.   25. An effective amount of the peptide according to claim 9 in a pharmaceutically acceptable delivery system. A vaccine composition comprising a mixture of immunogens.   26. 11. A pepti according to claim 10 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising a mixture of immunogens.   27. The pepti according to claim 11 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising a mixture of immunogens.   28. 13. A pepti according to claim 12 in an effective amount in a pharmaceutically acceptable delivery system. A vaccine composition comprising a mixture of immunogens.   29. Allele by administering an effective amount of the vaccine composition of claim 13. How to treat ghee reactions.   30. Allele by administering an effective amount of the vaccine composition of claim 14. How to treat ghee reactions.   31. Allele by administering an effective amount of the vaccine composition of claim 15. How to treat ghee reactions.   32. Allele by administering an effective amount of the vaccine composition of claim 16. How to treat ghee reactions.   33. Allele by administering an effective amount of the vaccine composition of claim 17. How to treat ghee reactions.   34. Allele by administering an effective amount of the vaccine composition of claim 18. How to treat ghee reactions.   35. Allele by administering an effective amount of the vaccine composition of claim 19. How to treat ghee reactions.   36. An allele by administering an effective amount of the vaccine composition of claim 20. How to treat ghee reactions.   37. Allele by administering an effective amount of the vaccine composition of claim 21. How to treat ghee reactions.   38. Allele by administering an effective amount of the vaccine composition of claim 22 How to treat ghee reaction.   39. Allele by administering an effective amount of the vaccine composition of claim 23. How to treat ghee reactions.   40. An allele by administering an effective amount of the vaccine composition of claim 24. How to treat ghee reactions.   41. An allele by administering an effective amount of the vaccine composition of claim 25. How to treat ghee reactions.   42. An allele by administering an effective amount of the vaccine composition of claim 26. How to treat ghee reactions.   43. An allele by administering an effective amount of the vaccine composition of claim 27. How to treat ghee reactions.   44. An allele by administering an effective amount of the vaccine composition of claim 28. How to treat ghee reactions.