Methods of production of autologous t cells for treatment of b-cell malignancies and other cancers and compositions thereof

A cell population and cell technology, applied in the field of T cells, can solve the problems of difficult commercial application and time-consuming

Pending Publication Date: 2022-04-26
凯德药业公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The process for producing autologous engineered T cells for use in cancer therapy is time-consuming (10-24 days), involves two cycles of retroviral transduction, and is poorly suited for commercial application (see Kochenderfer et al., Blood .2012119:2709-2720; Johnson et al., Blood.2009;114(3):535-546)

Method used

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  • Methods of production of autologous t cells for treatment of b-cell malignancies and other cancers and compositions thereof
  • Methods of production of autologous t cells for treatment of b-cell malignancies and other cancers and compositions thereof
  • Methods of production of autologous t cells for treatment of b-cell malignancies and other cancers and compositions thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Preparation of ex vivo genetically modified autologous cells

[0066] figure 1 A schematic diagram of an exemplary T cell production process ("improved" process) according to one embodiment is provided. This improved process also includes improvements to the traditionally used process for producing T cells (the "previous" process) (see the figure 2 ), while maintaining T cell product characteristics. Specifically, the improved process is a closed process that surprisingly avoids the use of serum. Moreover, this improved process uses a single cycle of transduction to produce a population of transduced T cells. Furthermore, cells subjected to a total expansion time of 6 days showed a more naïve immunophenotypic profile when using this procedure compared to cells subjected to an expansion time of 10 days. This process enables reproducible production of products with target numbers of transduced T cells expressing a chimeric antigen receptor (CAR) such as CD...

Embodiment 2

[0080] Example 2: Growth performance of T cells expanded in cell culture bags

[0081]Embodiments described herein provide a method for efficient production of engineered autologous T cell therapy within 6 days. The following improvements have been achieved over the existing technology: the process time has been shortened to 6 days instead of the 24, 14 or 10 days previously used (this reduces the number of tests (including RCR tests) required for product release); T cell products, including a higher proportion of naive T cells, achieve increased potency and effectiveness; larger numbers of cells can be used to initiate culture to compensate for shorter manufacturing times; closed systems are used to perform the methods described in the present invention identification of serum-free culture conditions that support T cell growth; single-cycle retroviral transduction in culture bags; cell culture activation and expansion in culture bags rather than vials; The product. These im...

Embodiment 3

[0101] Example 3: Development of transduction conditions in a closed system

[0102] Previously, transduction of PBMCs was performed in 6-well plates treated with non-tissue culture. These plates were incubated at 2-8°C with 10 μg / mL of Coating was performed overnight, or at room temperature for 2 hours. After incubation, remove The plates were blocked with 2.5% HSA for 30 minutes, followed by washing with HBSS+5mM HEPES. In the plate-based process, retroviral vectors are applied to coated wells and spun down in a centrifuge, followed by removal of approximately 75% of the viral supernatant, followed by addition of cells by spinnoculation for transduction .

[0103]In the present invention, three studies were performed to optimize the method for transducing PBMC in closed cell culture bags concentrations and to determine whether HSA washing and virus supernatant removal affected transduction. The first experiment was in Origen PermaLife TM PL07 culture bag, in which...

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Abstract

The present invention relates to the field of T cells and provides methods of producing autologous T cells and compositions thereof for the treatment of B-cell malignancies and other cancers. A method of making T cells expressing a cell surface receptor that recognizes a specific antigen moiety on the surface of a target cell, the method comprising enriching a population of lymphocytes; stimulating the population of lymphocytes with one or more T cell stimulators to produce a population of activated T cells, the stimulation being performed in a closed system using a serum-free culture medium; transduction of the activated population of T cells with a viral vector comprising a nucleic acid molecule encoding a cell surface receptor, producing a transduction population of T cells using single cycle transduction, the transduction being performed in a closed system using a serum-free culture medium; the transduced population of T cells is expanded for a predetermined time resulting in an engineered population of T cells, the expansion being performed in a closed system using a serum-free culture medium. The methods and processes described herein can be completed in significantly shorter time.

Description

[0001] divisional application [0002] The present invention is a divisional application of a Chinese patent application filed on February 4, 2015, entitled "Production method and composition of autologous T cells for treating B-cell malignancies and other cancers", application number 201680038907.4. technical field [0003] The invention relates to the field of T cells, and provides a method for producing T cells expressing cell surface receptors, an engineered T cell population expressing cell surface receptors, a pharmaceutical composition and a method for producing T cells. Background technique [0004] The process for producing autologous engineered T cells for use in cancer therapy is time-consuming (10-24 days), involves two cycles of retroviral transduction, and is poorly suited for commercial application (see Kochenderfer et al., Blood . 2012 119:2709-2720; Johnson et al., Blood. 2009; 114(3):535-546). Therefore, it is desirable to develop improved methods of the T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/62C12N15/12C12N15/867A61K39/00A61P35/00
CPCC12N2500/90C12N2501/2302C12N2501/515C12N2510/00A61K38/1774C12N5/0636A61P35/00C07K2319/03C07K14/7051A61K39/4611A61K39/4631A61K39/464412A61K35/17A61K39/0011A61K39/39558A61K2039/5156A61K2039/5158C07K16/2896C07K2319/74
Inventor 马克·贝特尔史蒂芬·A·费德曼史蒂芬·A·罗森伯格
Owner 凯德药业公司
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