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Modular antigen transporter molecules (MAT molecules) for modulating immune reactions, associated constructs, methods and uses

A modular, antigen-based technology, applied in chemical instruments and methods, biochemical equipment and methods, antibody medical components, etc., can solve problems such as allergic reactions and desensitization

Inactive Publication Date: 2010-05-12
BOEHRINGER LNGELHEIM VETMEDICA GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Although it is possible and intentional to use antigens as cargo molecules, it has so far not been possible to generate in an individual an immune response sufficient to achieve specific requirements in terms of targeting and dosage, e. Require

Method used

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  • Modular antigen transporter molecules (MAT molecules) for modulating immune reactions, associated constructs, methods and uses
  • Modular antigen transporter molecules (MAT molecules) for modulating immune reactions, associated constructs, methods and uses
  • Modular antigen transporter molecules (MAT molecules) for modulating immune reactions, associated constructs, methods and uses

Examples

Experimental program
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Effect test

example 28

[0093] spacer module

[0094] Spacer modules that can be used in the present invention are all molecules that are suitable for coupling other modules that are constituents of the MAT molecule to each other. Coupling can be done both covalently and non-covalently. The role of the spacer module is to enable The various modules of the MAT molecule are spatially separated from each other such that they do not adversely affect each other on their respective functions. The modules of the MAT molecule of the invention can be coupled via spacer modules which can then be chemically or enzymatically reacted And cleave again, for example by protease. In this way, the modules of the MAT molecule connected by spacer modules can be separated from each other again when required.

[0095] In general, all currently known or future known proteases can be used in the present invention [29, 30]. Currently commonly used proteases are thrombin, factor Xa, enterokinase or the TAGZyme system (Qiagen...

Embodiment 1

[0158] Example 1: Cloning of expression vectors for MAT molecules

[0159] All molecular biology methods described below were performed according to standard methods known to those skilled in the art [43]. Vector pQE-30 (Qiagen, Hilden, Germany) was used to clone the vector for expression of MAT molecule (modular antigen transport molecule).

[0160] In the first step, the nucleic acid sequence encoding the transport module is introduced into a bacterial expression vector. The DNA sequence encoding the amino acid GYGRKKRRQRRR of HIV Tat was introduced into vector pQE-30 in the form of a synthetic oligonucleotide. In addition to the HIV Tat sequence, the oligonucleotide contains a recognition sequence for the restriction enzyme Bgl II at the 5' end and recognition sequences for BamH I, Spe I, Pst I and Hind III at the 3' end. Subsequently, the synthesized HIV Tat sequence was digested with Bgl II and Hind III, and the vector pQE-30 was digested with restriction endonucleases ...

Embodiment 2

[0161] Embodiment 2: obtain the coding sequence of antigen Bet v 1, Asp f 1, Asp f 6 and Der p 1

[0162] The coding sequences for the various antigens in the antigen moiety were isolated by various methods known to those skilled in the art [43]. The Bet v 1 sequence was obtained by synthetic oligonucleotides, the Asp f1 and Asp f6 sequences had been obtained in previous studies and were isolated from the vectors used in these studies [45,46] and subsequently introduced into pQE-30 Vector, Der p 1 sequence was isolated by reverse transcriptase PCR using mRNA from the house dust mite (Dermatophagoides pteronyss inus).

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Abstract

By adding the novel developed modular antigen transporter molecules (MAT molecules) and other instruments related thereto, the immune system of an individual can be effectively modulated in a targetedmanner. The MAT molecules contain a translocation module which transports the MAT molecule inside the cell, an active intracellular targeting molecule resulting in intracellular transport of the MATmolecule to the organelles of the cell which are responsible for antigen processing and an antigen module which determines the specificity of the immune response modulated by the MAT molecule. Optionally, other modules such as tag modules or spacer modules can be contained in the MAT molecule. The invention comprises the inventive MAT molecules, antibodies which are produced using said MAT molecules and therapeutic agents and diagnostic reagents which are produced with MAT molecules. The invention also comprises applications containing said MAT molecules, antibodies, therapeutic agents and diagnostic reagents.

Description

technical field [0001] The present invention relates to stimulating and suppressing the immune system to prevent, treat or diagnose diseases associated with under- or over-activation of the immune system, including infectious diseases, neoplastic diseases, allergies, autoimmune diseases, graft rejection React and so on. At the heart of the invention is a novel method of influencing the immune system through a MAT molecule consisting of at least three components: [0002] 1. A transport module (module), its effect is to make the MAT molecule penetrate into the cell from the outside of the cell, [0003] 2. An intracellular targeting module that acts to cause the MAT molecule to be processed intracellularly in order to produce an altered immune response or altered antigen presentation, and [0004] 3. An antigenic moiety that determines the specificity of the modulated immune response. [0005] The combination of these three elements produces a MAT molecule, whereby the immun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62A61K39/00A61P39/00A61P37/00A61P31/00A61P35/00C07K14/38
CPCC07K2319/40C07K14/38C07K2319/10A61K39/00C12N2810/40C12N15/62C12N2310/111A61P29/00A61P31/00A61P35/00A61P37/00A61P37/04A61P37/06A61P37/08A61P39/00
Inventor 诺贝特·兰平克拉默瑞·雷托扎比内·弗吕克格尔伊莎贝尔·戴格尔
Owner BOEHRINGER LNGELHEIM VETMEDICA GMBH
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