EP0811016A1 - Immunogenes a base de peptide synthetique pour le traitement de l'allergie - Google Patents

Immunogenes a base de peptide synthetique pour le traitement de l'allergie

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Publication number
EP0811016A1
EP0811016A1 EP95914876A EP95914876A EP0811016A1 EP 0811016 A1 EP0811016 A1 EP 0811016A1 EP 95914876 A EP95914876 A EP 95914876A EP 95914876 A EP95914876 A EP 95914876A EP 0811016 A1 EP0811016 A1 EP 0811016A1
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EP
European Patent Office
Prior art keywords
gly
lys
phe
val
peptide
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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EP95914876A
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German (de)
English (en)
Other versions
EP0811016A4 (fr
Inventor
Chang Yi Wang
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United Biomedical Inc
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United Biomedical Inc
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Publication of EP0811016A1 publication Critical patent/EP0811016A1/fr
Publication of EP0811016A4 publication Critical patent/EP0811016A4/en
Withdrawn legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4283—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an allotypic or isotypic determinant on Ig
    • C07K16/4291—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an allotypic or isotypic determinant on Ig against IgE
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/64—Medicinal preparations containing antigens or antibodies characterised by the architecture of the carrier-antigen complex, e.g. repetition of carrier-antigen units
    • A61K2039/645—Dendrimers; Multiple antigen peptides

Definitions

  • the present invention relates to the use of a composition of a synthetic peptide, in a linear or radially branching multimeric form, as an immunogen for eliciting the production in healthy mammals, including humans, of high titer antibodies to the effector site on the CH4 domain of the e-chain of the human IgE heavy- chain, and to the use of the composition as a vaccine to provide an immunotherapy for the treatment of allergy.
  • Immunotherapy for the prevention of IgE-mediated allergic responses has been by desensitization or hyposensitization, wherein a gradually increasing amount of an allergen is given to a patient to reduce the effects of subsequent exposure to that allergen' 11 .
  • Limitations to such an allergen-based immunotherapy include difficulties in identifying the allergen involved and, if an allergen is identified, the adverse reactions frequently caused by the use of the identified allergen' 2 ' .
  • drugs to block the cascade of cellular events that is responsible for allergic reactions.
  • These drugs include anti-histamines, decongestants, ⁇ 2 agonists, and corticosteroids.
  • Anti-histamines, decongestants, and ⁇ 2 agonists act on events downstream of IgE in the allergic cascade, making them palliative remedies which address only the allergy symptoms.
  • Preventative treatments must act on cellular events closer to the initiation of IgE- mediated allergic reactions.
  • These palliatives provide relief that is short term and partial.
  • the relief of symptoms is frequently accompanied by adverse side effects, e.g. anti-histamines may cause restlessness or drowsiness, and ⁇ 2 agonists have sometimes been associated with increased morbidity in asthmatic patients.
  • Corticosteroids are powerful immunosuppressants and are highly efficacious for the treatment of allergic symptoms. However, they stimulate adverse hormonal activities and may cause an undesirably broad immunosuppression.
  • IgE immunoglobulin-like kinase
  • This may be accomplished either by suppressing IgE synthesis, such as is achieved by the inconvenient desensitization method; or by blocking the process by which IgE-allergen complexes stimulate the degranulation of mast cells and basophils with the concomitant release of the chemical mediators of hypersensitivity.
  • Anti-IgE CH4 peptide antibodies derived from immunizations with e chain-related "peptide-carrier protein conjugates” were also used for structure action studies on the degranulation of IgE-sensitized cells, by observing inhibitory activities' 5,11,12 ' .
  • a rabbit anti-peptide serum selected on the basis of its better-than-average anti-peptide titer, reduced the decapeptide-induced histamine release from rat peritoneal mast cells in a titer-dependent fashion.
  • This inhibitory activity was further confirmed by in vivo tests in a rat passive cutaneous anaphylaxis (PCA) model system.
  • PCA passive cutaneous anaphylaxis
  • small peptides are poor immunogens. To make small peptides immunogenic, they are usually joined to large carrier proteins by chemical conjugation or by gene fusion. These processes, however, generally produce unpredictable conformational changes in a peptide. Further, the immune response is frequently misdirected to the immunodominant carrier. Consequently, the development of a potent vaccine to provide long-lasting relief from allergies awaits further immunogen design.
  • the structurally active IgE CH4 decapeptide sequence in the human IgE CH4 domain is double underlined (SEQ ID NO:l) .
  • the amino acid code used in the Table is: A, alanine; R, arginine; N, asparagine; D, aspartic acid; C, cysteine; Q, glutamine;
  • E glutamic 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.
  • an objective of the present invention to employ a group of IgE e-chain based peptide immunogens chemically synthesized in either a radially branching form or a linear T helper epitope containing form, to elicit high titer antibodies to the decapeptide effector site of the CH4 domain of the human e-chain, when introduced to mammals, including humans.
  • Another objective is to design optimal peptide immunogens, with specific amino acid sequences taken from the human IgE heavy chain CH4 domain (IgE CH4) attached to peptides containing promiscuous human helper T cell epitopes in a specific orientation which, when introduced into mammals, including humans, will stimulate production of high titers of efficacious antibodies to the effector site on human IgE CH4.
  • These antibodies should inhibit mast cell activation, reduce the release of chemical mediators such as histamines that are responsible for allergy symptoms, depress IgE-mediated passive cutaneous anaphylaxis (PCA) reaction, and suppress allergen-induced IgE production by B lymphocytes.
  • PCA passive cutaneous anaphylaxis
  • a further objective is to develop an effective IgE e-chain peptide-based vaccine, employing compositions containing such branching multimeric or linear immunogens, to provide immunotherapy for the treatment of allergic reactions.
  • peptide immunogens are made by solid phase synthesis.
  • the peptide immunogens comprise a series of radially branched multimeric peptides containing a ten amino acid IgE CH4 peptide (SEQ ID NO:l) , or an immunogenic analog thereof; a series of multimeric branched peptides containing the IgE CH4 peptide (SEQ ID NO:l) or an immunogenic analog thereof together with a helper T-cell epitope (Th epitope) ; and a series of linear monomeric peptides containing the IgE CH4 peptide (SEQ ID N0:1) or an immunogenic analog thereof together with a portion of a helper T-cell epitope (Th epitope) .
  • the IgE CH4 peptide is taken from the Fc region of the IgE heavy chain, i.e. e-chain CH4 domain (IgE CH4) .
  • IgE CH4 e-chain CH4 domain
  • the linear peptides are preferred. Compositions containing these peptides are used to immunize healthy mammals, e.g. guinea pigs, rats, and humans, to elicit the production of high titer antisera specific for the IgE CH4 effector site (SEQ ID N0:1) and free of irrelevant antibodies.
  • vaccines containing the synthetic peptides as the key immunogen may also be prepared with an effective amount of a multimeric- branching peptide or a linear peptide in the presence of a proper adjuvant and/or delivery vehicle. It is expected that such vaccine compositions will elicit a more focused anti-IgE peptide response than those of the peptide- carrier protein conjugates currently used by Stanworth et al. (1 ) , thus providing a better immunotherapy for the treatment of allergy.
  • This invention is directed to the use of a novel group of peptide-based immunogens for the generation of high titer antibodies to an effector site on the CH4 domain of human IgE e heavy chain (SEQ ID NO:l) in healthy mammals, including humans, for the treatment of IgE- mediated allergic diseases.
  • allergy symptoms the immediate result of IgE-dependent hypersensitivities, are caused by chemical mediators released by mast cells and basophils.
  • the release is triggered when a mast cell or basophil that has been sensitized with surface-bound IgE binds to an allergen for which the surface-bound IgE is specific.
  • the triggering is actuated by the binding of the allergen to the Fab' portion of the surf ce-bound IgE in an antigen-antibody type interaction.
  • the allergen/antibody binding crosslinks the bivalent surface- bound IgE and induces a conformational change in the distal Fc region of IgE, the region of IgE in direct contact with a high affinity Fc receptor on the cell surface.
  • the conformational change activates the cell-IgE-allergen complex with the resultant release of mediators, including histamine, by the cell.
  • mediators including histamine
  • Effector site(s) on IgE are believed to participate in the triggering event.
  • the presence of specific anti-IgE antibodies directed against such "effector sites”, through either active or passive immunization, may lead to inhibition of the cell activation process in hosts suffering from allergic reactions by interfering with the interaction between the IgE "effector sites" and the cell surface.
  • Such interventions through the use of specific anti-IgE antibodies i.e. a kind of immunotherapy, can be achieved either passively, through prophylactic treatment with specific "site-directed" antibodies to IgE, or, more preferably, actively, by providing the host with a vaccine comprised of site-directed peptide immunogens, to elicit the production by the host of its own site-directed anti- IgE antibodies. It is believed that active immunization will provide a more effective and longer lasting protection.
  • specific immunogens are provided wherein synthetic immune stimulatory elements are linked to the CH4 decapeptide of IgE (SEQ ID NO:l) in a specific orientation such that potent antibodies directed to this effector site on IgE can be broadly generated in a genetically diverse host population.
  • these antibodies block the stimulatory action of IgE on mast cells and basophils, thus resulting in an effective treatment to prevent IgE-mediated allergic diseases.
  • the peptide immunogens of the present invention are capable of eliciting antibodies with serological cross-reactivity with the target amino acid sequence of the Fc region of IgE (SEQ ID N0:1) while being substantially incapable of mediating non-cytolytic histamine release.
  • the initial dose e.g. 0.2-2.5 mg; preferably 1 mg, of immunogen is to be administered by injection, preferably intramuscular, followed by repeat (booster) doses.
  • Dosage will depend on the age, weight and general health of the patient as is well known in the therapeutic arts. While there is no particular limitation to the species of mammals suitable for the production of antibodies, it is generally preferred to use mice, rabbits, guinea pigs, pigs, goats, rats or sheep, etc. as the hosts.
  • immunogen relates to synthetic peptides which are capable of ⁇ inducing antibodies against the IgE CH4 decapeptide (SEQ ID N0:1), which antibodies lead to the suppression of IgE-mediated basophil and mast cell degranulation.
  • the immunogen of this invention included multimeric peptides or its analogs with a branching lysyl core matrix structure.
  • IgE CH4 decapeptide SEQ ID N0:1
  • the analogs of IgE CH4 decapeptide include the synthetic peptide analogs described by Stanworth et al. (3,4,5) , which are incorporated herein by reference.
  • the molecular weight of the immunogen should be higher than 5,000 and preferably be higher than 10,000.
  • the repeating branch unit for the peptide should be equal to or higher than 4.
  • Bifunctional amino acids such as lysine followed by attachment to an amino acid with a preferably non- charged side chain, such as Gly or Ala, are useful in the making of the core matrix structure.
  • the amino acid acts as a spacer in between the peptide branches to allow maximum freedom to attain the conformation necessary for optimal presentation.
  • the immunogen referred to in the present invention also included linear peptides which contain promiscuous helper T cell epitopes (Th epitopes) . These Th epitopes were covalently attached in a defined fashion to the decapeptide effector sequence (SEQ ID N0:1) , with or without a spacer, so as to be adjacent to the N terminus of the decapeptide, in order to evoke efficient antibody responses.
  • the immunogen may also be comprised of an immune stimulatory sequence corresponding, for example, to a domain of an invasin protein from the bacteria Yersinia spp (19) . The invasin domain may also be attached through a spacer to a Th epitope.
  • the "immunogen" of the present invention minimizes the generation of irrelevant antibodies, thus eliciting a more focused immune response to the "target sequence", i.e., the desired IgE CH4 cross-reactivity (SEQ ID NO:l) , without producing undesirable side effects which may complicate the immunotherapy process for the treatment of allergy.
  • target sequence i.e., the desired IgE CH4 cross-reactivity (SEQ ID NO:l)
  • SEQ ID NO:l the desired IgE CH4 cross-reactivity
  • a short target sequence such as the 10 amino acid IgE CH4 segment Lys-Thr-Lys-Gly-Ser-Gly- Phe-Phe-Val-Phe (SEQ ID N0:1)
  • a short peptide antigen is usually a T cell-dependent antigen, i.e.
  • the short IgE CH4 decapeptide (SEQ ID NO:l) or an immunogenic analog thereof does not contain a T helper cell epitope.
  • the branched multimeric and linear immunogens comprising the short IgE CH4 decapeptide are designed herein to provide for artificially built-in functional helper T-cell epitopes.
  • the peptides immunogens of this invention are represented by the formula (A) n -(Th) ra -(B) 0 -(IgE CH4 peptide) p wherein
  • A is an amino acid, ⁇ -NH 2 , a fatty acid, a derivative of a fatty acid, or an invasin domain;
  • B is an amino acid;
  • Th is a helper T cell epitope or an immune enhancing analog or segment thereof;
  • IgE CH4 peptide is Lys-Thr-Lys-Gly-Ser-Gly-Phe-
  • Phe-Val-Phe (SEQ ID NO:l) or an immunogenic analog thereof; n is from 1 to 10; m is from 1 to 4; o is from 0 to 10; and p is from 1 to 3.
  • the peptide immunogens of the present invention comprise from about 20 to about 100 amino acid residues, preferably from about 20 to about 50 amino acid residues and more preferably from about 20 to about 35 amino acid residues.
  • A is an amino acid, it can be any non- naturally occurring or any naturally occurring amino acid.
  • Non-naturally occurring amino acids include, but are not limited to, ⁇ -alanine, ornithine, norleucine, norvaline, hydroxyproline, thyroxine, ⁇ -amino butyric acid, homoserine, citrulline and the like.
  • Naturally-occurring amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine.
  • n is greater than one, and two or more of the A groups are amino acids, then each amino acid is independently the same or different.
  • A When A is a fatty acid, such as stearic acid or palmitic acid or a fatty acid derivative, such as a tripalmitoyl cysteine (Pam 3 Cys) group, it acts as an adjuvant by enhancing the immunostimulating properties of the Th epitope (20) .
  • A When A is a fatty acid or its derivative it is usually located at the amino terminus of the peptide. Furthermore, when one of A is a fatty acid, there are 2 or 3 additional amino acid A moieties.
  • the fatty acids useful in the invention have a hydrocarbon chain of 8 to 24 carbon atoms which may be saturated or unsaturated.
  • A When A is an invasin domain, it is an immune stimulatory epitope from the invasin protein of a Yersinia species. This immune stimulatory property results from the capability of this invasin domain to interact with the ⁇ l integrin molecules present on T cells, particularly activated immune or memory T cells.
  • the specific sequence for an invasin domain found to interact with the ⁇ l integrins has been described by Brett et al (19> .
  • the invasin domain (Inv) for linkage to a promiscuous Th epitope has the sequence:
  • Thr-Tyr-Gln-Phe (SEQ ID NO: 25) or is an immune stimulatory analog thereof from the corresponding region in another Yersinia species invasin protein.
  • Such analogs may contain substitutions, deletions or insertions to accommodate strain to strain variation, provided that the analogs retain immune stimulatory properties.
  • n is 4 and A is ⁇ .-NH 2 , lysine, lysine and lysine in that order. In another embodiment n is 1 and A is ⁇ -NH 2 . In yet another embodiment, n is 4 and A is o.-NH 2 , an invasin domain (Inv) , glycine and glycine in that order.
  • Inv invasin domain
  • B comprises naturally occurring or the non- naturally occurring amino acids as described above. Each B may be independently the same or different.
  • B is lysine
  • a branched polymer can be formed. For example, if o is 7 and all seven B groups are lysine then a branching K core (K 4 K 2 K) is formed when the peptide synthesis is conducted without protecting the lysyl side chain e-amino group.
  • Peptides with a K core have eight branch arms, with each branch arm being identical and represented as 11 (A) n - (Th) m - M or "(IgE CH4 peptide with built-in-Th) -" .
  • the amino acids of B can form a flexible hinge, or spacer, to enhance the immune response to the Th epitope and IgE CH4 decapeptide or an analog thereof.
  • Examples of sequences encoding flexible hinges can be found in the immunoglobulin heavy chain hinge region. Flexible hinge sequences are often proline rich. One particularly useful flexible hinge is provided by the sequence Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:24) , where Xaa is any amino acid, preferably aspartic acid. Immunogenicity can also be improved through the addition of spacer residues (e.g. Gly-Gly) between the promiscuous Th epitope and the IgE CH4 decapeptide or an analog thereof.
  • spacer residues e.g. Gly-Gly
  • Th is a Th epitope comprising natural or non- natural amino acids.
  • Th epitope may consist of a continuous or discontinuous epitope; not every amino acid of Th is necessarily part of the epitope.
  • Th epitopes, including analogs and segments thereof, to be suitable for the present invention are capable of enhancing or stimulating an immune response to the IgE CH4 decapeptide (SEQ ID NO:l) or an analog thereof.
  • Th epitopes that are immunodominant and promiscuous are highly and broadly reactive in animal and human populations with widely divergent MHC types' 21"23 '.
  • the Th domain suitable for the present invention has from about 10 to about 50 amino acids and preferably from about 10 to about 30 amino acids. When multiple Th epitopes are present (i.e. m ⁇ 2) , then each Th epitope may be independently the same or different.
  • Th epitope analogs include substitutions, additions, deletions and insertions of from one to about 10 amino acid residues in the Th epitope.
  • Th segments are contiguous portions of a Th epitope that are sufficient to enhance or stimulate an immune response to the IgE CH4 decapeptide (SEQ ID NO:l) or an analog thereof.
  • Th epitopes of the present invention include hepatitis B surface and core antigen helper T cell epitopes (HB s Th and HB c Th) , pertussis toxin helper T cell epitopes (PT Th) , tetanus toxin helper T cell epitopes (TT Th) , measles virus F protein helper T cell epitopes (MV F Th) , Chlamydia trachomatis major outer membrane protein helper T cell epitopes.
  • HB s Th and HB c Th hepatitis B surface and core antigen helper T cell epitopes
  • PT Th pertussis toxin helper T cell epitopes
  • TT Th tetanus toxin helper T cell epitopes
  • MV F Th measles virus F protein helper T cell epitopes
  • Chlamydia trachomatis major outer membrane protein helper T cell epitopes
  • Th epitope sequences are provided below:
  • Ser-Leu-Asp SEQ ID NO: 5
  • PTi Th Lys-Lys-Leu-Arg-Arg-Leu-Leu-Tyr-Met-Ile-Tyr-Met-
  • TTi Th Lys-Lys-Gln-Tyr-Ile-Lys-Ala-Asn-Ser-Lys-Phe-Ile-
  • PT 1A Th Tyr-Met-Ser-Gly-Leu-Ala-Val-Arg-Val-His-Val-Ser- Lys-Glu-Glu (SEQ ID NO: 9)
  • MV F1 Th Ser-Glu-Ile-Lys-Gly-Val-Ile-Val-His-Arg-Leu-Glu- Gly (SEQ ID NO: 12) and Leu-Ser-Glu-Ile-Lys-Gly-Val-Ile-Val-His-Arg-Leu- Glu-Gly-Val (SEQ ID NO: 61)
  • CT Th Ala-Leu-Asn-Ile-Trp-Asp-Arg-Phe-Asp-Val-Phe-Cys-
  • TraT 2 Th Gly-Leu-Ala-Ala-Gly-Leu-Val-Gly-Met-Ala-Ala-Asp- Ala-Met-Val-Glu-Asp-Val-Asn (SEQ ID NO: 35)
  • TraT 3 Th Ser-Thr-Glu-Thr-Gly-Asn-Gln-His-His-Tyr-Gln-Thr- Arg-Val-Val-Ser-Asn-Ala-Asn-Lys (SEQ ID NO.-36)
  • the Th epitope is HB S Th, P1 1 Th, PT 2 Th, TT X Th, TT 3 Th, or MV F1 Th.
  • the Th epitope is covalently attached through spacer B to the N terminus of the IgE CH4 decapeptide (SEQ ID NO:l) .
  • the IgE CH4 peptide is Lys- Thr-Lys-Gly-Ser-Gly-Phe-Phe-Val-Phe (SEQ ID NO:l) , a decapeptide.
  • the IgE CH4 peptide may be replaced by an immunogenic analog.
  • the immunogenic analogs thereof may contain a substitution, addition, deletion, or insertion of from one to about four amino acid residues provided that the analog is capable of eliciting an immune response crossreactive with the IgE CH4 decapeptide (SEQ ID NO:l) .
  • the substitutions, additions, and insertions may be made with natural or non-natural amino acids as defined herein.
  • Immunogenic analogs of the IgE CH4 peptide (SEQ NO:l) have been identified by Stanworth et al. (3,4,5) and are incorporated herein by reference.
  • preferred peptide immunogens of this invention are monomeric peptides containing IgE CH4 decapeptide (SEQ ID NO:l) or an immunogenic analog thereof and Th. More specifically, preferred peptide immunogens are those linear constructs containing IgE CH4 (SEQ ID NO:l) or an immunogenic analog thereof; a spacer (e.g Gly- Gly) ; a Th epitope selected from the group consisting HB S Th, PTi Th, PT 2 Th, TT X Th, TT 3 Th, and MV F1 Th (SEQ ID NOS:5, 6,11,7,10, 61, respectively) ; and optionally the Inv domain (SEQ ID NO:25) .
  • Preferred peptide immunogen compositions include, for example, Peptide Nos. 19-23 and 28 (Tables 5 and 6, SEQ ID NOS:51-55, 62) .
  • the peptide immunogens of this invention may be made by chemical synthesis well known to the ordinarily skilled artisan. See, for example, Grant, ed. Synthetic Peptides' 24 ' .
  • peptide immunogens may be synthesized using the automated Merrifield techniques of solid phase synthesis with the o.-NH 2 protected by either t-Boc or F- moc chemistry using side chain protected amino acids on, for example, an Applied Biosystems Peptide Synthesizer Model 430A or 431.
  • Di- ⁇ , e (t-Boc) lysine residues are used in place of t-Boc lysine with a 2,4- dichlorobenzyl protecting e-amino group.
  • A When A is a fatty acid, it may be added easily to the N-terminus of the resin bound peptide by the well known carbodiimide method.
  • the lipoamino acid S- [2,3-Bis (palmitoyloxy) - (2R) -propyl-N-palmitoyl- (R) - cysteine (Pam 3 Cys) is chemically synthesized.
  • Pam 3 Cys may then be coupled to the N terminus of a peptide by solid- phase synthesis using Pam 3 Cys-OH in the final coupling step to link the lipoamino acid to a resin-bound peptide chain.
  • the solid-phase peptide can be elongated with additional serine and lysine residues at the N-terminus.
  • the resin After complete assembly of the desired peptide immunogen, the resin is treated according to standard procedures to cleave the peptide from the resin and deblock the functional groups on the amino acid side chains.
  • the free peptide is purified by HPLC and characterized biochemically, for example, by amino acid analysis or by sequencing. Purification and characterization methods for peptides are well known to one of ordinary skill in the art.
  • Th-IgE CH4 decapeptide constructs of the invention include the ligation of the haloacetylated and the cysteinyl peptide through the formation of a "thioether" linkage.
  • cysteine can be added to the C terminus of a Th- containing peptide and the thiol group of cysteine is used to form a covalent bond to an electrophilic group such as an N ⁇ chloroacetyl-modified or a maleimide-derivatized - or e-NH 2 group of a lysine residue that is attached to the N-terminus of the IgE CH4 decapeptide (ID SEQ N0:1) or an immunogenic analog thereof.
  • the subject peptides can also be polymerized. Polymerization can be accomplished for example by reaction between glutaraldehyde and the -NH 2 groups of the lysine residues using routine methodology.
  • the linear "A-Th- spacer-IgECH4" peptide constructs (e.g., Peptide Nos. 19- 23 and 28, SEQ ID NOS:51-55 and 62) may also be polymerized -or co-polymerized by utilization of an additional cysteine added to the N-terminus of the linear "A-Th-spacer-IgECH4" construct.
  • the thiol group of the N- terminal cys.teine may be used for the formation of a "thioether" bond with a halochloroacetyl-modified or a maleimide-derivatized cn- or e-NH 2 group of a lysine residue that is attached to the N-terminus of a branched poly-lysyl core molecule (e.g., K 2 K, K 4 K 2 K, K 8 K 4 K 2 K) .
  • the longer linear peptide immunogens may be synthesized by well known recombinant DNA techniques. Any standard manual on DNA technology provides detailed protocols to produce the peptides of the invention.
  • a gene encoding a peptide of this invention the amino acid sequence is reverse translated into a nucleic acid sequence, and preferably using optimized codon usage for the organism in which the gene will be expressed.
  • a synthetic gene is made, typically by synthesizing overlapping oligonucleotides which encode the peptide and any regulatory elements, if necessary.
  • the synthetic gene is inserted in a suitable cloning vector and recombinants are obtained and characterized.
  • the peptide is then expressed under suitable conditions appropriate for the selected expression system and host.
  • the peptide is purified and characterized by standard methods.
  • the efficacy of the peptide immunogen of the present invention may be established by injecting the immunogen into an animal, and then monitoring the humoral immune response to IgE CH4 decapeptide (SEQ ID N0:1) or an immunogenic analog thereof, as detailed in the Examples.
  • Suitable animals include mice, rats, rabbits, guinea pigs, pigs, goats, sheep, or the like.
  • Another aspect of this invention provides a vaccine composition comprising an effective amount of one or more of the peptide immunogens of this invention in a pharmaceutically acceptable delivery system.
  • Such vaccine compositions are used for prevention of atopic allergic reactions including allergic rhinitis, those of food allergies, asthma, anaphylaxis, and other IgE-mediated hypersens ' itive reactions such as virally-induced asthma.
  • the subject peptide immunogens can be formulated as a vaccine composition using adjuvants, pharmaceutically-acceptable carriers or other ingredients routinely provided in vaccine compositions.
  • Such formulations are readily determined by one of ordinary skill in the art and include formulations for immediate release and/or for sustained release, and for induction of systemic immunity and/or induction of localized mucosal immunity, which may be accomplished by, for example, immunogen entrapment by microparticles.
  • the formulation may also include adjuvants or emulsifiers such as alum, incomplete Freund's adjuvant, liposyn, saponin, squalene, L121, emulsigen and ISA 720 and the like.
  • the vaccine of the present invention may be administered by any convenient route including subcutaneous, oral, intramuscular, or other parenteral or enteral route. It may be administered as a single dose or in multiple doses. Immunization schedules are readily determined by the ordinarily skilled artisan.
  • the vaccine compositions of the instant invention contain an effective amount of one or more of the synthetic peptide immunogens containing the IgE CH4 decapeptide or its immunogenic analog and a pharmaceutically acceptable carrier.
  • the dosage unit form may be formulated to contain about 0.5 ⁇ g to about 1 mg of each peptide per kg body weight . When delivered in multiple doses, the effective dose may be conveniently divided to contain the appropriate amounts per unit dosage form.
  • the vaccine compositions of the present invention may be formulated to contain a cocktail of two or more of the subject peptide immunogens to enhance immunoefficacy in a broader population and thus provide a better immune response against IgE CH4 decapeptide.
  • a cocktail of Peptide Nos. 19, 20, 21, 23, and 4 is useful.
  • the composition may also be formulated to comprise lipopeptides to provide a built-in adjuvant.
  • the immune response to synthetic IgE CH4 decapeptide- containing immunogens may also be improved by delivery through entrapment in or on biodegradable microparticles of the type described by O'Hagan et al i25) .
  • the immunogens can be encapsulated with or without adjuvant, including covalently attached Pam 3 Cys, and such microparticles may carry an immune stimulatory adjuvant such as Freund's Incomplete Adjuvant or alum.
  • the microparticles function to potentiate immune responses to the immunogen, including localized mucosal immunity. Such localized immunity is especially desirable, for example, for mucosally localized allergic reactions.
  • Vaccine compositions in microparticular form may also be formulated to provide time-controlled release for sustained or periodic responses, for oral administration, and for topical administration' 25"26 ' . Examples of specific peptide immunogens are provided herebelow to illustrate the present invention and are to be used to limit the scope of the invention.
  • the synthesis of the multimeric peptides proceeds by the limited sequential propagation of a trifunctional amino acid to serve as a low molecular weight matrix core is the basis for the formation of a branching multimeric peptide antigen system.
  • the trifunctional amino acid, Boc-Lys (Boc) , or di- (Boc) -Lys is most suitable since both N"- and N e - amino acid groups are available as reactive ends. Thus, sequential propagation of di- (Boc) -Lys will generate 2 n reactive ends.
  • the first coupling of di- (Boc) -Lys onto a solid phase resin will produce two reactive amino ends to bind two peptide chains.
  • Sequential generations of a second, third, and fourth step with di- (Bpc) -Lys will therefore generate respectively tetravalent, octavalent, and hexadecavalent ends for binding multimeric peptide chains antigens.
  • Such multimeric peptides are useful as immunogens.
  • Branched octameric Peptide Nos. 1 and 2 as described above were synthesized for use as immunogens.
  • the branched antigens contain a small heptalysyl core surrounded by a layer of high density of uniform peptide- antigens around the core matrix.
  • This design differs from the conventional peptide-carrier conjugate antigen which contains a large protein carrier such as PPD or KLH and a small peptide antigen randomly distributed on the surface of the protein carrier in many undefined forms.
  • octameric peptide immunogens employs a combination of Boc-amino acid resin- bound benzhydrylamide and tBoc-chemistry.
  • Boc-amino acid resin- bound benzhydrylamide For example, an 8-branched heptalysyl core resin was prepared by coupling di-t-Boc Lys onto an extra low loading of 0.14 mmole/g MBHA (4-methylbenzhydrylamine) resin on a Biosearch 9500 instrument. Two coupling cycles of di- (Boc) -Lys for each was followed by two capping reacticns using 0.3 M acetylimidazole in DMF dimethylformamide.
  • Acid-labile tert-butyloxycarbonyl (t-Boc) was used for the protection of N-oc amino acid.
  • the following functional side-chain.protecting groups were used: O- benzyl for Thr, Ser, Glu and Tyr; N ⁇ -tosyl for Arg; BOM, i.e., BOC-N im -Benzyloxymethyl for His, N e - dichlorobenzyloxycarbonyl for Lys; S-4-methylbenzyl- for Cys; O-cyclohexyl for Asp and CHO for Trp.
  • CFA complete Freund's adjuvant
  • Anti-peptide antibody activity is determined by ELISA (enzyme-linked immunosorbentassay) using 96-well flat bottom microtiter plates which were coated with the corresponding immunogen. Aliquots (100 ⁇ L) of a peptide immunogen solution at a concentration of 5 ⁇ g/mL were incubated for 1 hour at 37°C. The plates were blocked by another incubation at 37°C for 1 hour with a 3% gelatin/PBS solution. The blocked plates were then dried and used for the assay. Aliquots (100 ⁇ L) of the test guinea pig sera, starting with a 1:10 dilution in a sample dilution buffer and ten-fold serial dilutions thereafter, were added to the peptide coated plates. The plates were incubated for 1 hour at 37°C. Normal guinea pig serum was used as a control.
  • ELISA enzyme-linked immunosorbentassay
  • Immuno ⁇ en preparation Peptide immunogens A-H (Table 3) are synthesized by solid phase synthesis using F-moc chemistry on an Applied Biosystems Peptide Synthesizer Model 430A or 431 according to manufacturer's instructions. After complete assembly of the peptide, the resin is treated according to standard procedures to cleave the peptide from the resin and deprotect the functional groups on amino acid side chains.
  • Peptide immunogen A is a linear peptide with three domains: 3 lysine residues (3K) , the hepatitis B surface antigen helper T cell epitope (HB s Th epitope) and IgE CH4 peptide.
  • Peptide immunogen A is thus represented as 3K-HB s Th-IgE CH4 peptide.
  • the actual sequences for Peptide immunogen A and for Peptide immunogens B-H are shown in Table 5 (SEQ ID NOS:15-22) .
  • each peptide immunogen is dissolved and combined with an adjuvant solution (Complete Freund's Adjuvant, Incomplete Freund's Adjuvant, or 0.2% Alum) to result in a final concentration of 0.5 mg/ml.
  • the solution is stored at 4°C until use and vortexed for 3 to 5 min prior to injection.
  • Each rat receives 100 ⁇ g per injection.
  • Booster injections are given s.c. at weeks 2 and 5. Blood is collected at weeks 3, 6, 7 and 11.
  • Blood collection from the middle caudal artery is performed following anesthesia of the rats by intraperitoneal injection of 1 L of sodium pentobarbital
  • Blood is collected immediately into a 5 mL syringe outfitted with a 23 gauge needle. Typically, 2 to 2.5 mL of blood is obtained. The serum is collected by centrifugation for 25 min at 3000 rpm. The serum is aliquoted in 300 ⁇ L volumes and stored frozen until used for ELISA assays.
  • Linear peptide immunogens represented as A-Th- GG-IgE CH4, where A may be either NH 2 -, Lys-Lys (2K) , Lys- Lys-Lys (3K) , or an invasin domain (Inv) (SEQ ID NO:25), Th is a T helper peptide, GG is a Gly-Gly spacer, and IgE CH4 is the target decapeptide (SEQ ID NO:l) , are synthesized as described in Example 3. These peptide immunogens are shown in Table 4 as Peptide Immunogens Nos. 4-17 (SEQ ID NOS:37-50) . The synthesized and cleaved peptides are used to immunize rats to test for efficacy.
  • Immunogen Peptide Nos. 4-17 (1 per trial)
  • Dose 100 ⁇ g per immunization Route: intramuscular
  • Adjuvant Freund's Complete/Incomplete
  • Dose Schedule week 0 (FCA) , 3 and 6 weeks (IFA)
  • ELISA for anti-peptide activity solid-phase immunosorbent is monomeric Peptide No. 3 of the IgE CH4 decapeptide sequence (SEQ ID NO:1) .
  • Blood is collected, processed into serum, and stored prior to titering by ELISA as described in Example 2, with the exception of using horseradish peroxidase- labelled goat anti-rat IgG antibody instead of goat anti- guinea pig IgG as the tracer.
  • Peptide immunogens Nos. 18-23 (ID SEQ ID NOS:51-56) as shown in Table 5, were synthesized as described in Example 3.
  • the formula for peptide immunogens Nos. 18-23 may be represented as A-Th-GG-IgECH4, wherein A is either the N terminus, Lys-Lys (2K) , Lys-Lys-Lys (3K) , or the invasin domain (Inv) (SEQ ID NO:25) separated from the Th sequence by a spacer GG; Th is selected from the group consisting of HB S Th, PT X Th, PT 2 Th, MV F1 Th, or TT 3 Th; GG is a Gly-Gly spacer; and IgECH4 is the IgE CH4 decapeptide (SEQ ID NO:l) .
  • Immunogen Peptide Nos. 18-28 (1 per group) (SEQ ID NOS:51-60 and 62)
  • Blood was collected, processed into serum, and stored prior to titering by ELISA as described in Example 2 with the exception of substituting horseradish peroxidase- labelled goat anti-rat IgG antibody for anti-guinea pig IgG as the tracer. All sera were assayed by anti-peptide ELISA and those samples which gave A 492nm values of ⁇ 0.2 at a 1:100 dilution were recorded as seropositive.
  • the immunopotencies of Peptide immunogens Nos. 18-28 were evaluated by the anti-peptide ELISA and are shown in Table 6 as the number of rats in each group of 4 or 5 that converted to seropositive reactivity for IgE CH4 Peptide No. 3 on weeks 6 and 8 (i.e., A 492ram ⁇ 0.2 at a 1:100 dilution) , in response to the experimental immunizations.
  • the peptide immunogens of this Example of polarity Th-GG-IgECH4 (Peptide Nos. 18-23 and 28, SEQ ID NOS:51-56 and 62) showed significant efficacy for the induction of antibodies to the IgE CH4 decapeptide (Peptide No. 3, SEQ ID NO:l) .
  • All 6 groups of rats immunized with the peptide immunogens of this polarity (Peptide Nos. 18-23, 28) showed significant conversion to seropositivity compared to the control.
  • Prevalences of seroconversion for the groups varied from 1/5 to 5/5 by week 6 and seroconversion prevalences continued to increase between weeks 6 and 8 in response to the third dose of immunogens.
  • Example 3-5 Establishing the relative efficacies of the many different linear constructs containing IgE CH4 decapeptide and Th (Examples 3-5) permits selection of useful peptide immunogens to formulate a cocktail vaccine composition.
  • Individual Th-GG-IgECH4 constructs carrying immunodominant promiscuous Th peptides derived from measles virus F protein, tetanus toxin and pertussis toxin (Peptide Nos. 19-23) were proven by the study of Example 5 to be efficacious in eliciting antibody responses to the IgECH4 decapeptide (SEQ ID N0:1) .
  • a formulation containing a mixture of these linear peptides may provide a desired maximum immunogenicity in a genetically diverse population.
  • Blood is collected, processed into serum, and stored prior to titering by ELISA as described in Example 5.
  • This experiment is designed to demonstrate improved performance of the immunogens of the present invention as compared to the known immunogens of the prior art 114,15 '.
  • the results are useful for the evaluation of two mixtures of efficacious peptide immunogens, each containing three Th peptides, demonstrate the usefulness of the immune stimulatory Inv domain (cocktail 2 contains Inv, cocktail 1 does not) , and the efficacy of the adjuvant, Alum, in a vaccine composition of the invention.
  • Th peptides are derived from children's vaccines, childhood vaccinations are a potential source of Th memory in an immunized human population. Thus, children's vaccines have the potential to afford enhanced immunopotency to anti-allergy vaccines comprised of mixtures of such Th peptides.
  • the clinical protocol below has been designed to demonstrate efficacy for compositions of the invention formulated as a mixture of such linear "A-Th-Spacer-IgE Ch4 decapeptide" peptide immunogens, in a widely acceptable adjuvant, Alum.
  • Hay fever patients Season & Duration: Hay fever seasons, 8 weeks
  • Dose Molar equivalents of each synthetic peptide to equal 500 ⁇ g or 125 ⁇ g of peptide per dose Route: intramuscular Dose Schedule: week 0, and 4 weeks Evaluation schedule: weeks 0, 4, and 6
  • Blood is collected, processed into serum, and stored prior to titering by ELISA as described in Example 5.
  • Efficacy and safety of the vaccine composition "cocktail 1" are evaluated serologically, by skin reaction tests, the rate of patient usage of hay fever medication, physical examination of patients for allergic symptoms and adverse reactions, and interviewing the patients to obtain their subjective assessments of the effect of using the products.
  • Serological evaluations include the aforementioned ELISA for antipeptide titer, and a standard automated spectrofluorimetric assay to determine reduction in histamine levels 115 ' as well as to ascertain that the products do not trigger histamine release.
  • the skin test is an intradermal test in which a standardized solution of allergens is injected into the upper layers of the skin.
  • Reactions to the allergens are quantitated by determining the area of the typical "wheal and flare" produced in response to the allergens.
  • the expected results include significant improvement in allergic symptoms at the endpoint of the study, and no evidence of histamine release triggered by the vaccine composition of the invention.
  • Passively-sensitized human basophils are used in a well- known histamine-release assay for an in vitro evaluation of antibodies induced by immunizations with IgE CH4 decapeptide immunogens.
  • Human basophils are prepared from the venous blood of volunteers and then passively sensitized with IgE specific for benzylpenicilloyl-human serum albumin conjugate (BPO-HSA) that is prepared from the blood of donors hyperimmunoglobulemic for BPO-HSA- specific IgE. Histamine release by the sensitized basophils is affected by the addition of either BPO-HSA or IgE CH4 Peptide No. 3 (SEQ ID NO:l) .
  • the basophils Prior to the addition of the agents to induce histamine release, the basophils are combined with serial dilutions of antiserum to IgE CH4 decapeptide (SEQ ID NO:l) or pre-immune control serum. Samples are analyzed for histamine release by the automated fluorescence technique. The percentage of histamine release is calculated from the ratio of sample to total basophil histamine content after spontaneous release is subtracted from both 127 ' . The capacity of the experimental antiserum to inhibit histamine release is demonstration of in vitro efficacy.
  • MOLECULE TYPE Poylpeptide e-chain of human IgE
  • Val Cys Ser Arg Asp Phe Thr Pro Pro Thr Val Lys lie Leu Gin 5 10 15
  • MOLECULE TYPE Polypeptide .e-chain of rat IgE (x) PUBLICATION INFORMATION:
  • Val Thr Trp Val Arg Glu Arg Lys Lys Ser lie Gly Ser Ala Ser 80 85 90
  • MOLECULE TYPE Polypeptide e-chain of mouse IgE (x) PUBLICATION INFORMATION:
  • 200 205 210 lie Thr Leu Pro Gin Val Ser Gin Arg Ser Ala Pro Glu Val Tyr
  • Trp Leu Gly Asp Gly Lys Leu lie Ser Asn Ser Gin His Ser Thr
  • MOLECULE TYPE peptide
  • SEQUENCE DESCRIPTION SEQ ID NO: 6 :
  • Lys Lys Leu Arg Arg Leu Leu Tyr Met lie Tyr Met Ser Gly Leu 1 5 10 15
  • Lys Lys Gin Tyr lie Lys Ala Asn Ser Lys Phe lie Gly lie Thr 1 5 10 15
  • MOLECULE TYPE peptide
  • SEQUENCE DESCRIPTION SEQ ID NO:10: Tyr Asp Pro Asn Tyr Leu Arg Thr Asp Ser Asp Lys Asp Arg Phe 1 5 10 15
  • MOLECULE TYPE linking group
  • MOLECULE TYPE peptide (xi ) SEQUENCE DESCRIPTION : SEQ ID NO : 34 :

Abstract

L'invention concerne un procédé permettant d'induire la production, chez les mammifères en bonne santé dont les humains, des anticorps à titre élevé dirigés contre un site effecteur dans une chaîne lourde d'IgE humain, tel qu'un site dans le domaine CH4 de la chaîne ⊂, au moyen de compositions d'immunogènes de peptide synthétique sous une forme multimère ramifiée radialement (telle que des peptides octamères ou hexadécamères) ou monomère disposée linéairement, pour inhiber l'activation des mastocytes et réduire la production d'IgE induite par un allergène. L'invention porte également sur l'utilisation de ces immunogènes de conception optimale relatifs à la chaîne ⊂ d'IgE et exempts de protéines porteuses en tant que composants déterminants dans un vaccin synthétique destiné à produire une immunothérapie pour le traitement de l'allergie. Les peptides selon l'invention contiennent des séquences de stimulation immunitaire contenant un déterminant antigénique du lymphocyte T auxiliaire intégré lié en tandem dans un sens spécifique, pour faciliter la stimulation de la réponse immunitaire dirigée contre le domaine CH4 d'IgE.
EP95914876A 1994-03-28 1995-03-24 Immunogenes a base de peptide synthetique pour le traitement de l'allergie Withdrawn EP0811016A1 (fr)

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US21846194A 1994-03-28 1994-03-28
US218461 1994-03-28
US32891294A 1994-10-25 1994-10-25
PCT/US1995/003741 WO1995026365A1 (fr) 1994-03-28 1995-03-24 Immunogenes a base de peptide synthetique pour le traitement de l'allergie
US328912 2002-12-23

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2790442A1 (fr) 2013-04-09 2014-10-15 Alcatel Lucent Système de commande, appareil, procédés et support de stockage lisible par ordinateur stockant des instructions pour un n'ud de réseau et/ou un contrôleur de réseau

Families Citing this family (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5843462A (en) 1995-11-30 1998-12-01 Regents Of The University Of Minnesota Diphtheria toxin epitopes
ATE230417T1 (de) 1996-03-01 2003-01-15 Novartis Erfind Verwalt Gmbh Peptid-immunogene zur schutzimpfung gegen und behandlung von allergien
GB9611673D0 (en) * 1996-06-05 1996-08-07 Peptide Therapeutics Ltd Meningococcal vaccine
CA2315537A1 (fr) 1997-12-16 1999-06-24 Regents Of The University Of Minnesota Procedes permettant de traiter des reponses immunitaires indesirables
TWI227241B (en) 1998-06-20 2005-02-01 United Biomedical Inc IgE-CH3 domain antigen peptide, peptide conjugate containing the same, and pharmaceutical composition for treating allergies containing the peptide conjugate
US6090388A (en) * 1998-06-20 2000-07-18 United Biomedical Inc. Peptide composition for prevention and treatment of HIV infection and immune disorders
US6913749B2 (en) 1998-11-02 2005-07-05 Resistentia Pharmaceuticals Ab Immunogenic polypeptides for inducing anti-self IgE responses
KR20020084841A (ko) 1999-04-23 2002-11-11 파멕사 에이/에스 Ｉｌ5 활성을 다운-조절하는 방법
GB9913327D0 (en) * 1999-06-08 1999-08-11 Smithkline Beecham Biolog Vaccine
US6846486B1 (en) 2000-02-24 2005-01-25 Advanced Biotherapy Concepts, Inc. Method of treating allergy by administering an anti-histamine antibody
WO2001069253A2 (fr) * 2000-03-15 2001-09-20 Heska Corporation MODELE TRIDIMENSIONNEL D'UN COMPLEXE ENTRE UNE CHAINE ALPHA DU RECEPTEUR Fc-EPSILON ET UNE REGION Fc D'UN ANTICORPS IgE ET LEURS UTILISATIONS
AU7658101A (en) * 2000-07-28 2002-02-13 Cytos Biotechnology Ag Compositions for inducing self-specific anti-ige antibodies and uses thereof
CA2421274A1 (fr) * 2000-09-06 2002-03-14 Pharmexa A/S Procede de regulation negative d'ige
US7265208B2 (en) 2001-05-01 2007-09-04 The Regents Of The University Of California Fusion molecules and treatment of IgE-mediated allergic diseases
JP2003047482A (ja) 2001-05-22 2003-02-18 Pfizer Prod Inc 非アナフィラキシー誘発性ＩｇＥワクチン
US7604955B2 (en) 2001-08-13 2009-10-20 Swey-Shen Alex Chen Immunoglobulin E vaccines and methods of use thereof
IL158287A0 (en) 2003-10-07 2004-05-12 Yeda Res & Dev Antibodies to nik, their preparation and use
WO2009023816A1 (fr) * 2007-08-15 2009-02-19 Emory University Procédés de préparation d'anticorps monoclonaux utilisant la technologie du transfert adoptif d'épitope de peptide de fusion (f-peat)
CA2717930A1 (fr) * 2008-03-13 2009-09-17 Yossef Kliger Nouveaux peptides derives de gp96
EP2788021B1 (fr) 2011-12-09 2017-01-18 Bavarian Nordic A/S Vecteur du poxvirus codant des antigènes bactériens lié à le fragment c de la toxine tétanique
US9187553B2 (en) * 2012-01-25 2015-11-17 Swey-Shen Chen Displaying native human IgE neutralizing FcepsilonRIa-contacting IgE B-cell epitopes by constraining super beta(b)-strands and cystine knots on thermostable protein scaffold
US9657071B2 (en) * 2012-06-18 2017-05-23 Nippon Zenyaku Kogyo Co., Ltd. IgE peptide vaccine
CN112237638B (zh) * 2020-10-28 2022-11-29 西安华牧生物科技有限责任公司 降低放射性核素肾浓聚的组合探针及其制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994021676A1 (fr) * 1993-03-24 1994-09-29 The Regents Of The University Of California ISOFORMES IgE ET LEURS METHODES D'UTILISATION
WO1994025060A1 (fr) * 1993-04-27 1994-11-10 Ladd Anna E Peptides immunogenes de la lhrh et stimulators d'immunite universelle de synthese pour vaccins
WO1995010532A1 (fr) * 1993-10-11 1995-04-20 Peptide Therapeutics Limited Composes utilises dans les traitements anti-allergies
WO1996006357A1 (fr) * 1994-08-24 1996-02-29 Peptide Therapeutics Limited Criblage pour des composes a potentiel therapeutique

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8913737D0 (en) * 1989-06-15 1989-08-02 Univ Birmingham A novel anti-allergy treatment

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994021676A1 (fr) * 1993-03-24 1994-09-29 The Regents Of The University Of California ISOFORMES IgE ET LEURS METHODES D'UTILISATION
WO1994025060A1 (fr) * 1993-04-27 1994-11-10 Ladd Anna E Peptides immunogenes de la lhrh et stimulators d'immunite universelle de synthese pour vaccins
WO1995010532A1 (fr) * 1993-10-11 1995-04-20 Peptide Therapeutics Limited Composes utilises dans les traitements anti-allergies
WO1996006357A1 (fr) * 1994-08-24 1996-02-29 Peptide Therapeutics Limited Criblage pour des composes a potentiel therapeutique

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO9526365A1 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2790442A1 (fr) 2013-04-09 2014-10-15 Alcatel Lucent Système de commande, appareil, procédés et support de stockage lisible par ordinateur stockant des instructions pour un n'ud de réseau et/ou un contrôleur de réseau

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EP0811016A4 (fr) 1997-12-10
CA2186595A1 (fr) 1995-10-05

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