Artificial T helper cell epitopes as immune stimulators for synthetic peptide immunogens

A peptide immunogen and helper cell technology, applied in the field of peptide immunogen

Inactive Publication Date: 2005-05-18
SHANGHAI SHEN LIAN BIOMEDICAL CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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Method used

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Examples

Experimental program
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Effect test

example

[0088] The peptide immunogens of the invention can be prepared by chemical methods known to those skilled in the art. See, eg, Fields et al., "Synthetic Peptides: A User's Manual," Chapter 3, Grant, W.H. ed., Freeman & Co., New York, NY, 1992, p.77. The peptide can be synthesized using automated Merrifield solid-phase peptide synthesis using t-Boc or Fmoc protection of α-NH2 or side chain amino acids. Equipment for peptide synthesis is commercially available. An example is the Model 430A or 431 Applied Biosystems Peptide Synthesizer.

[0089] After assembly of the desired peptide immunogen is complete, 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. Free peptides are purified by HPLC and biochemically characterized by, for example, amino acid analysis, by sequencing, or by mass spectrometry. Methods of purifying and characterizing peptides are well known to those of o...

Embodiment 1

[0110] Immunization of rats with LHRH-containing peptide immunogen

[0111] The peptides listed in Tables 2a and 2b were synthesized and tested as described below.

[0112] A. Peptide synthesis. The peptides listed in Tables 2a and 2b, respectively, were synthesized with Fmoc chemistry by Merrifield solid-phase synthesis technique on an Applied Biosystems automated peptide synthesizer (Models 430, 431 and 433A). A peptide construct comprising a constructed synthetic antigen library (SSAL), eg, an artificial Th site designated as SEQ ID NO:3, is prepared by providing a mixture of desired amino acids selected for a particular position. After assembly of the desired peptide or combined peptides, the resin should be treated with trifluoroacetic acid to cleave the peptide from the resin and deblock the protecting groups on the amino acid side chains, following standard procedures.

[0113] The excised, extracted and washed peptides were purified by HPLC and characteriz...

Embodiment 2

[0134] LHRH peptide mixtures induce broader immune castration in rats

[0135] Confirming the relative effectiveness of the various artificial TH epitope / LHRH constructs shown in Example 1, a more potent mixture of peptides was selected to combine into enhanced immunogenicity. The Th / LHRH peptide immunogen mixture was more potent than any of the individual peptides in the mixture of US 5795551. Moreover, a mixture of individual constructs carrying promiscuous Th epitopes derived from MVF Th (SEQ ID NO: 1) and HbsAg Th (SEQ ID NOS: 23-25) provided a greater advantage in genetically diverse populations than those derived from only one Broader responses to peptide compositions of Th epitopes intermingling Th epitopes. Therefore, a peptide composition containing a mixture of peptides of the invention derived from MVF Th and HbsAg Th was assembled and the effect of this mixture was tested and compared to a composition containing the individual peptides of the mixture.

[0...

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PUM

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Abstract

The present invention is directed to novel artificial T helper cell epitopes (Th epitopes) designed to provide optimum immunogenicity when used in peptide immunogens comprising B cell epitopes or peptide haptens, a target antigenic site of a target antigen for eliciting antibodies thereto. The artificial Th epitopes are covalently linked to the target antigenic site and optionally an immunostimulatory sequence to provide effective and safe peptide immunogens.

Description

field of invention [0001] The present invention relates to a peptide immunogen comprising a novel artificial T helper cell epitope (Th) covalently linked to a desired target antigenic site, including a B cell epitope, and optionally a Conventional immunostimulatory sequence. Artificial Th epitopes endow peptide immunogens with the ability to induce strong T helper cell-mediated immune responses and generate antibodies directed against the "antigenic site of interest". The present invention also provides favorable substitution of carrier proteins and pathogen-derived T helper epitopes in peptide immunogens developed using novel artificial T helper epitopes to improve immunogenicity. [0002] Several rules have been developed in predicting the amino acid sequence of T cell epitopes. However, because there is no central unified theory of how or how to prepare a specific amino acid sequence that can be used as a T-cell epitope, the established rules are empirical and not general...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/00A61K39/00A61K39/015A61K39/135A61K39/21A61K39/39A61P9/10A61P15/00A61P31/04A61P31/12A61P31/18A61P33/06A61P35/00A61P37/04A61P37/08C07K1/04C07K7/08C07K7/23C07K14/09C07K14/16C07K14/445C07K14/47C07K14/655C07K14/73C07K16/00C07K19/00
CPCA61K39/00C07K7/23C07K14/005C07K14/445C07K14/47C07K14/655C07K14/70514C07K16/00C07K19/00C12N2740/16122C12N2770/32122C07K2317/52A61P15/00A61P31/04A61P31/12A61P31/18A61P33/06A61P35/00A61P37/02A61P37/04A61P37/08A61P9/10Y02A50/30
Inventor 王长怡
Owner SHANGHAI SHEN LIAN BIOMEDICAL CORP
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