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Targeted immunogens

A technology for immunogens and tumor antigens, applied in specific peptides, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problems of ineffective entry of exogenous peptides, limited immunogenicity, etc.

Inactive Publication Date: 2007-02-28
圣诺菲·帕斯图尔公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Despite the many advantages of peptide-based vaccines (safety, ease of production), they show limited immunogenicity
This is partly due to inefficient access of exogenous peptides to the MHC class I presentation pathway

Method used

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  • Targeted immunogens
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0154] Preparation of immunogenic target peptide

[0155] All peptides were synthesized by Bio-Synthesis Incorporated (Lewisville, Texas) using standard techniques.

[0156] To prove the feasibility of the epitope conjugation system, cytotoxic T lymphocyte (CTL) epitopes were conjugated to various transduction sequences. Choose the following transcytosis peptides for attachment to the epitope:

[0157] TAT: GYGRKKRRQRRR

[0158] hPER1-1: SRRHHCRSKAKRSRHH

[0159] hPER1-2: RRHHRRSKAKRSR

[0160] AntPHD: RQIKIWFQNRRMKWKK

[0161] Certain epitope peptides are linked to transcytosis sequences with linker sequences. The linker is selected from a sequence found in nature directly at the N-terminus of the epitope sequence, or is selected based on known immunological parameters. The selected linker sequence is shown below:

[0162] OVA: LEQLE (natural)

[0163] DEVWEL (synthetic)

[0164] NP 366-374: RGVQI (natural)

[0165] gp100(154-162): FVYVW (natural)

[0166] Cho...

Embodiment 2

[0202] Immunological test

[0203] A. The hPER1-CTL epitope conjugate can form a CTL target structure when incubated with cells in vitro.

[0204] In order to determine whether hPER1-CTL conjugates can form a CTL target structure, the 51 Cr-labeled RMA cells with 10 -11 g / ml NP peptide (ASNENMETM) or hPER1-NP peptide (RRHHRRSKAKRSRASNENMETM) pulsed or left untreated (no peptide) and incubated at 37°C for 1 hour. The cells were then washed and tested for CTL recognition in a standard 4-hour chromium release assay using T cells obtained from the spleen of C57BL / 6 mice immunized with influenza virus. Figure 1A demonstrates that RMA target cells can be sensitized to CTL-mediated lysis when incubated with 10 pg / ml hPER1-NP peptide.

[0205] In addition, 51 Cr labeled P815-A2 / K b 10 for cells -6 g / ml 280-9V peptide (YLEPGPVTV) or hPER1-280-9V (RRHHRRSKKAKRSRYLEPGPVTV) pulsed or left untreated (no peptide) and incubated at 37°C for 1 hour. The cells were then washed and tested for CTL re...

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PUM

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Abstract

The present invention provides reagents and methods for producing and utilizing targeted immunogens. In preferred embodiments, an immunogen is conjugated to an amino acid sequence that targets the immunogen to the MHC presentation pathway. Using the reagents and methods provided herein, immunization protocols may be enhanced resulting in increased immunity of the host.

Description

[0001] Related application [0002] This application claims the priority of U.S. Provisional Application No. 60 / 533,728 filed on December 31, 2003. Invention field [0003] The present invention relates to reagents and methods for improving immunization protocols. For example, the amino acid sequence that directs the immunogenic amino acid sequence to the MHC presentation pathway. Background of the invention [0004] Although peptide-based vaccines have many advantages (safe, easy to produce), they show limited immunogenicity. This is partly due to the inability of foreign peptides to effectively enter the MHC class I presentation pathway. Thus, strategies that can enhance the delivery of peptides to MHC have the potential to increase the efficacy of peptide-based vaccines. One strategy is to link immunogenic sequences to "protein transduction domains" (PTD), which have been shown to drive the transport of proteins and peptides across cell membranes. Exemplary PTDs include HIT-Tat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/48C07K19/00C07K14/47
CPCA61K2039/627A61K2039/6031C07K14/47A61P31/00A61P35/00A61P37/02C07K19/00
Inventor A·R·昂格尔D·萨尔哈S·加利钱
Owner 圣诺菲·帕斯图尔公司