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Modified immune cells co-expressing chimeric antigen receptor and IL-6 antagonist for reducing toxicity and uses thereof in adoptive cell therapy

A technology of immune cells and IL-6, applied in genetically modified cells, blood/immune system cells, for targeting specific cell fusion, etc., can solve problems such as limitations of adoptive cell transfer therapy

Pending Publication Date: 2021-11-19
HUNAN SIWEIKANG THERAPEUTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Promise of adoptive cell transfer therapy is often limited by toxicities such as cytokine-related toxicity

Method used

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  • Modified immune cells co-expressing chimeric antigen receptor and IL-6 antagonist for reducing toxicity and uses thereof in adoptive cell therapy
  • Modified immune cells co-expressing chimeric antigen receptor and IL-6 antagonist for reducing toxicity and uses thereof in adoptive cell therapy
  • Modified immune cells co-expressing chimeric antigen receptor and IL-6 antagonist for reducing toxicity and uses thereof in adoptive cell therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0221] Example 1 : The role of IL-6 antagonistic antibodies expressed in 293T cells in inhibiting IL-6 signaling

[0222] HEK293T cells were transfected with the third-generation self-inactivating (SIN) lentiviral transfer vector of Lipofectamine 2000 (Thermo Scientific), which encodes single-chain variable fragments derived from reference antibodies 1, 2, 3 and 4 disclosed herein ( scFv) antibodies targeting IL-6 or IL-6R. The CD8 leader sequence precedes the anti-IL6 scFv. See description in Example 7 below. scFv antibody fused to the Fc fragment of human IgG1. Supernatants of transfected cells containing scFv antibodies expressed by transfected HEK293T cells were collected, diluted, and added to HEK-Blue IL-6 reporter cells (Invivogen) in the presence of 2 ng / ml human IL-6. HEK-Blue IL-6 reporter cells were used because of their ability to produce secreted embryonic alkaline phosphatase (SEAP) upon stimulation with human IL-6. After overnight incubation, the supernata...

Embodiment 2

[0224] Example 2 : Combination of anti-IL-6 antibody and IL-1RA expressed in 293T cells in blocking IL-6 and IL-1 signaling

[0225] The nucleic acids encoding Construct 1, Construct 2 and Construct 3 were cloned into a third-generation self-inactivating (SIN) lentiviral transfer vector. Construct 1 comprised a T2A linker from N-terminus to C-terminus, a scFv antibody (targeting IL-6) derived from reference antibody 2, a P2A linker and an IL-1 receptor antagonist (IL-1RA) (T2A-Sir-P2A -IL1RA). Construct 2 contains T2A linker, scFv antibody, (G 4 S) 3 Linker and IL1RA(T2A-Sir-(G4S) 3 -IL1RA). Construct 3 contains scFv antibody from N-terminus to C-terminus, (G 4 S) 3 Linker, IL1RA and T2A linker (Sir-(G 4 S) 3 -IL1RA-T2A). In constructs 1, 2 and 3, the CD8 leader sequence preceded the anti-IL6 scFv. See description in Example 7 below. In construct 1, the ahGH leader sequence is located between P2A and IL1RA.

[0226] 293T cells were transfected with the above-ment...

Embodiment 3

[0229] Example 3 : Combined effect of IL-6 and GM-CSF antagonistic antibodies expressed in 293T cells in blocking IL-6 and GM-CSF signaling

[0230] Constructs for expression of the three exemplary bispecific antibodies specific for IL-6 and GM-CSF, as well as IL1RA described in Example 2 above, were generated by conventional recombinant techniques. Each bispecific antibody contained scFv derived from reference antibody 2 (targeting IL-6) and scFv derived from reference antibody 7, reference antibody 8 or reference antibody 9 (all targeting GM-CSF). The two scFv fragments in the bispecific antibody are linked by a GSGGSG linker. Each bispecific antibody is linked to IL1RA via a P2A linker. These constructs were named Ab2 / Ab9-P2A-IL1RA, Ab2 / Ab7-P2A-IL1RA and Ab2 / Ab8-P2A-IL1RA (corresponding to Figure 3A to Figure 3C 1, 2 and 3 in ). In these constructs, the CD8 leader sequence preceded the anti-IL6 scFv, and the ahGH leader sequence was located between P2A and IL1RA. See...

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Abstract

A population of immune cells comprises modified immune cells co-expressing a chimeric antigen receptor and an IL-6 signaling antagonist (e.g., an anti-IL6 or anti-IL-6R antibody) and optionally an IL-1 signaling antagonist. Also provided herein are methods of producing such immune cell populations comprising the modified immune cells and methods of using such in cell therapy (e.g., to treat cancer, infectious diseases, or immune diseases).

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Application No. 62 / 789,311, filed January 7, 2019, U.S. Provisional Application No. 62 / 855,250, filed May 31, 2019, and U.S. Provisional Application No. 62 / 928,720, filed October 31, 2019 The entire content of each application is hereby incorporated by reference in its entirety for the benefit of its filing date. Background technique [0003] Adoptive cell transfer therapy is a type of immunotherapy that involves ex vivo expansion of autologous or allogeneic immune cells followed by infusion into the patient. Immune cells can be modified ex vivo to specifically target malignant cells. The promise of adoptive cell transfer therapy is often limited by toxicity, such as cytokine-related toxicity. For example, adoptive cell transfer immunotherapy may trigger a nonphysiological increase in cytokine levels (cytokine release syndrome), which may lead to death of the recipient (see, e.g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/00A61K39/395C07K16/00C07K16/46C07K19/00C12N5/10
CPCA61K39/395C07K2317/31C07K2317/76C07K2317/622C07K16/2866C07K16/2803C07K16/243C07K16/248C07K2319/03C07K2319/33C07K2319/32C07K2319/02C07K16/2878A61K31/7076A61K38/00A61K45/06A61K31/675C12N2510/00C12N5/0636C12N2501/2306C12N2501/2301C12N2501/599C07K14/7051A61P35/00A61K39/464412A61K2239/48A61K39/4631A61K2239/31A61K39/464417A61K2239/38A61K39/4611A61K2300/00C07K16/468
Inventor 胡璧梁
Owner HUNAN SIWEIKANG THERAPEUTICS CO LTD