Chimeric antigen receptors targeting tim-1

A chimeric antigen receptor, TIM-1 technology, applied in the field of preparation and use of such compositions, can solve problems such as not yet achieved success

Active Publication Date: 2019-11-22
CELDARA MEDICAL +1
View PDF62 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this approach has shown curative potential in many patients with chemotherapy-refractory hematological malignancies (Kalos et al., SciTransl Med. 2011; 3:95ra73; Maus et al., Cancer Immunol Res. 2013; 1: 26-31; Porter et al., N Engl J Med.2011; 365:725-733), but similar success has not been achieved in solid tumors

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
  • Chimeric antigen receptors targeting tim-1
  • Chimeric antigen receptors targeting tim-1
  • Chimeric antigen receptors targeting tim-1

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0289] In some embodiments, the method of preparation includes the step of freezing (eg, cryopreserving) the cells before or after isolation, incubation, and / or engineering. In some embodiments, the step of freezing followed by thawing removes granulocytes, and to some extent monocytes, from the cell population. In some embodiments, cells are suspended in a freezing solution to remove plasma and platelets, eg, after a washing step. In some aspects, any of a variety of known freezing solutions and parameters can be used. One example involves the use of PBS containing 20% ​​DMSO and 8% human serum albumin (HSA), or other suitable cell freezing medium. Then it was diluted 1:1 with culture medium so that the final concentrations of DMSO and HSA were 10% and 4%, respectively. Cells were then frozen to -80°C at a rate of 1°C per minute and stored in the gas phase of liquid nitrogen tanks.

[0290] therapeutic application

[0291] Isolated cells obtained by the methods describe...

Embodiment 1

[0409] Example 1: Design and synthesis of anti-TIM-1 CAR variants

[0410] according to figure 1 General schematic for generation of anti-TIM-1 CAR. For the six variants, Figure 2A A more detailed schematic of the CAR construct is provided. The CAR structure is based on a second generation CAR format (Gacerez et al., J Cell Physiol, 2016 Dec;231(12):2590-8). There are six different TIM-1 -responsive single change variable fragments (scFv) in Hv-linker-Lv or Lv-linker-Hv orientation from anti-TIM1 hybridoma clones 1.29, 2.70.2 or 2.59.2 ( Figure 2B ). These anti-TIM-1 scFvs were formed by fusing the variable domains of the heavy chain (VH) and light chain (VL) domains to the following 15 amino acid glycine (G)-serine (S) linker: (G4S)3 Linker (SEQ ID NO:201), 3 repeats of GGGGS (SEQ ID NO:200). These were cloned individually in frame into the hinge domain containing CD28 (residues 135-152 of CD28 or SEQ ID NO:214), the transmembrane domain of CD28 (residues 153-179 of ...

Embodiment 2

[0417] Example 2: Generation of anti-TIM-1 CAR T cells

[0418] Cell culture and retroviral transduction: The retroviral stocks described above were used using optimized methods (Cubillos-Ruiz et al., Oncotarget 2010; 1:329-33; Huarte et al., Blood 2008; 112:1259-1268 Stephen et al., Immunity 2014;41:427-439), protocols and resources previously developed at Celdara Medical to transduce human T cells from healthy donors.

[0419] For some variants, cell culture, retroviral transduction and purification protocols are summarized in image 3 middle. The resulting transduced cells were analyzed by flow cytometry ( Figure 4 ). For the purification of some variants, the protocol is as follows. Human PBMCs from healthy donors (HemaCare) were the source of T cells for CAR transduction. Donor PBMCs were thawed, reconstituted in complete medium, and pelleted by centrifugation. Cells were then resuspended in complete medium and then activated by incubation with IL-2 and anti-CD3 fo...

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

No PUM Login to view more

Abstract

The invention provides chimeric antigen receptors (CARs) that specifically bind to the T-cell immunoglobulin and mucin domain 1 (TIM-1) protein. The invention further relates to modified immune cells,e.g., T or NK cells, comprising such CARs, CAR-encoding nucleic acids, CAR-encoding vectors, and methods of making such compositions. The invention further relates to methods for therapeutic use of these CARs and modified immune cells for the treatment of a condition, disorder, or disease associated with cells expressing TIM-1 (e.g., cancer).

Description

[0001] Related application open [0002] This application claims the benefit of U.S. Provisional Application Serial No. 62 / 445,976, filed January 13, 2017, which is hereby incorporated by reference in its entirety. [0003] sequence public [0004] This application includes as part of its disclosure the Biological Sequence Listing in a file named "56867o1000.txt" and 193,169 bytes in size, created on January 12, 2018, which is hereby incorporated by reference in its entirety enter. technical field [0005] The invention disclosed herein relates to chimeric antigen receptors (CARs) that bind to antigenic T cell immunoglobulin domain and mucin domain 1 (TIM-1 ) proteins and uses of such CARs. In particular, chimeric antigen receptors are provided comprising an antigen binding domain that binds TIM-1, a transmembrane domain, and one or more intracellular signaling domains. The nucleotide sequence encoding the anti-TIM-1 CAR construct and the amino acid sequence comprising the ...

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): C07K16/30C07K16/28C07K14/705C12N5/0783C12N5/078A61K39/395A61K35/17A61P35/00
CPCA61K35/17A61P35/00C07K14/70521C07K16/2803A61K2039/505A61K2039/5156A61K2039/5158C07K2317/622C07K2317/73C07K2319/03C07K2319/40A61K38/00C07K14/7051C07K16/30C12N15/85
Inventor L-Z·何H·C·小玛什T·凯勒T·A·斯塔德海姆J·M·穆拉德J·雷德尔
Owner CELDARA MEDICAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap