Unlock instant, AI-driven research and patent intelligence for your innovation.

A group of chimeric antigen receptors (CARS)

A technology of chimeric antigen receptors and antigens, applied in the direction of antibodies, antibody medical components, antibody mimics/scaffolds, etc., can solve problems such as the destruction of healthy tissues

Pending Publication Date: 2021-08-20
圣安娜儿童癌症研究中心 +1
View PDF20 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This fact leads to so-called on-target / off tumor toxicity, i.e. destruction of healthy tissue expressing the antigen, since tumor-associated antigens are always expressed on healthy cells

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A group of chimeric antigen receptors (CARS)
  • A group of chimeric antigen receptors (CARS)
  • A group of chimeric antigen receptors (CARS)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0388] Example 1: Generation of rcSso7d-based low-affinity single-domain binding moieties for CAR panels according to the invention

[0389] In a first example it is shown a strategy for generating an antigen-binding moiety with low affinity suitable for use as an antigen-binding moiety of a CAR panel according to the invention. Reduced charge Sso7d (rcSso7d) is a reduced charge version of a small (-7 kDa) DNA-binding protein from the archaea Sulfolobus solfataricus. The reduced charge minimizes non-specific binding, due to reduced electrostatic interactions. rcSso7d is a single-domain protein antigen-binding moiety with high thermal stability and monomeric behavior, and is thus an example suitable for binding to scaffolds. Starting with the well-characterized antigen-binding portion rcSso7d E11.4.1, which binds to human EGFR with a K of 19 nM d Combined with (Traxlmayr et al., JBiol Chem. 2016; 291(43):22496-22508), by performing an alanine scan in which we replaced all ami...

Embodiment 2

[0404] Example 2: Extracellular disulfide bonds formed by cysteines prevent avidity effects that fully utilize the AND gate function of the CAR group

[0405] Extracellular disulfide bonds formed by cysteines in the extracellular hinge region, eg CD8α, prevent the use of the avidity effect according to the invention. This is demonstrated in Example 2, where a low-affinity mutant of the binding moiety "E11.4.1G32A" of Example 1 is fused to the CAR signaling backbone with two extracellular cysteine ​​residues in the hinge region of CD8α (UniProt ID P01732, positions C164 and C181) were substituted or not substituted with serine residues, respectively. While cysteine-containing CAR variants ("Cys") efficiently triggered T cell activation in response to target cells, serine-containing variants ("Ser") did not or only weakly triggered T cells. Thus, this example illustrates the importance of preventing disulfide bond formation when generating CAR molecules suitable for use in CAR ...

Embodiment 3

[0419] Example 3: Single-chain variable fragments (scFv) can trigger clustering of CARs in cell membranes, thereby preventing avidity effects using combinations of specific recognition antigens

[0420] The third example shows that the integration of scFv-based binding moieties in CAR molecules can prevent avidity effects utilizing combinations of specific recognition antigens. Figure 4 Schematic representation of the CAR construct shown in A showing the CAR variants tested (4D5-5-8cys-BB-3z, 4D5-5-8ser-BB-3z, 4D5-5(split)-8ser-BB-FKBP (36V )-3z) design. In the example shown, scFv 4D5-5 directed against HER2 was used as the antigen binding moiety and incorporated into a monomeric ("Ser") or dimeric ("Cys") CAR signaling backbone. Figure 4 B shows CAR expression in primary T cells. The effective binding affinity of scFv 4D5-5 was reported as 1.1 μΜ (Liu et al., Cancer Res. 2015; 75(17):3596-3607), which is comparable to the affinity of E11.4.1-G32A. Jurkat T cells expressi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
affinityaaaaaaaaaa
affinityaaaaaaaaaa
affinityaaaaaaaaaa
Login to View More

Abstract

Disclosed is a group of chimeric antigen receptors (CARs) consisting of two, three or four CAR molecules, wherein each member of the group of CARs is different in its amino acid sequence from one another, and wherein each of the CAR molecules of the group comprise at least a transmembrane domain and an ectodomain, wherein the ectodomain comprises one or two antigen binding moieties and / or one or two binding sites to which other polypeptides each comprising at least an antigen binding moiety are able to bind; wherein the ectodomain of each CAR molecule of the group in its prevalent conformation is free of cysteine amino acid moieties which are able to form intermolecular disulphide bonds with other CAR molecules of the group, respectively, and wherein each CAR molecule of the group comprises at least one heterodimerization domain.

Description

technical field [0001] The present invention relates to panels of chimeric antigen receptors (CARs) consisting of two, three or four CAR molecules. Background technique [0002] Immunotherapy using CAR T cells, in which T cells are modified to express chimeric antigen receptors (CARs), is one of the most promising approaches in cancer treatment. The high potential of this therapeutic strategy has been demonstrated to date by the impressive clinical responses of patients with B-cell malignancies. However, further replication of this success to other tumors is presently prevented by several obstacles (Lim and June, Cell. 2017; 168(4):724; Labanieh et al., Nat Biomed Eng. 2018; 2:377-391 ). For example, current CAR T cells are CARs typically directed against a single defined tumor-associated antigen. This fact leads to so-called on-target / off tumor toxicity, ie destruction of healthy tissue expressing the antigen, since tumor-associated antigens are always expressed on health...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00
CPCA61P35/00C07K2317/31C07K2318/20C07K2317/92C07K14/7051C07K2319/00A61K39/464412C12N5/0646A61K39/4611A61K39/464406A61K39/4631A61K2239/31A61K2239/48A61K39/464404C12N5/0636A61K2239/28A61K2239/38C07K16/2863C07K16/32C07K16/22C07K2317/13C07K2317/622C07K2319/02C07K2319/03C12N2510/00A61K35/17A61K38/00A61K2039/505C07K14/70503C07K14/70517C07K14/70578C07K14/71C07K14/7153C07K16/2803C07K2317/569C07K2317/76C07K2319/30C07K2319/33
Inventor B·扎尔策M·莱纳M·特拉克斯迈尔
Owner 圣安娜儿童癌症研究中心