US 12,252,545 B2
Therapeutic cell compositions and methods of manufacturing and use thereof
Daniel Getts, Stow, MA (US); and Yuxiao Wang, Belmont, MA (US)
Assigned to MYELOID THERAPEUTICS, INC., Cambridge, MA (US)
Filed by Myeloid Therapeutics, Inc., Cambridge, MA (US)
Filed on Mar. 15, 2021, as Appl. No. 17/202,018.
Application 17/202,018 is a continuation of application No. 16/826,708, filed on Mar. 23, 2020, granted, now 10,980,836.
Claims priority of provisional application 62/946,896, filed on Dec. 11, 2019.
Prior Publication US 2021/0361703 A1, Nov. 25, 2021
Int. Cl. C07K 16/28 (2006.01); A61K 39/00 (2006.01); A61K 40/17 (2025.01); A61K 40/24 (2025.01); A61K 40/31 (2025.01); A61K 40/42 (2025.01); A61P 35/00 (2006.01); C07K 14/705 (2006.01); C12N 5/0786 (2010.01); C12N 9/12 (2006.01); C12N 15/87 (2006.01)
CPC C07K 16/2896 (2013.01) [A61K 40/17 (2025.01); A61K 40/24 (2025.01); A61K 40/31 (2025.01); A61K 40/4205 (2025.01); A61K 40/4224 (2025.01); A61P 35/00 (2018.01); C07K 14/70578 (2013.01); C07K 16/2863 (2013.01); C12N 5/0645 (2013.01); C12N 9/1205 (2013.01); C12N 15/87 (2013.01); A61K 2239/38 (2023.05); A61K 2239/48 (2023.05); C07K 2317/52 (2013.01); C07K 2317/622 (2013.01); C07K 2319/03 (2013.01)] 17 Claims
 
1. A pharmaceutical composition comprising:
(a) a population of mRNA-electroporated cells lacking a viral component or a plasmid component, wherein the mRNA comprises a sequence encoding a chimeric fusion protein (CFP) or a sequence encoding an antigenic peptide, wherein:
(i) at least 50% of the cells in the population of mRNA-electroporated cells are CD 14+;
(ii) less than 10% of the cells in the population of mRNA-electroporated cells are dendritic cells; and
(iii)
(A) at least 50% of the cells in the population of mRNA-electroporated cells are CCR2+ (CD192+), and CCR5+ (CD195+) and two or more of: CD63+, CD56 CD120a+ (TNFR1+) or CD120b+ (TNFR2+);
(B) less than 50% of the cells in the population of mRNA-electroporated cells express CD64 CD68, CD80, CD86, CD163, CD206, CD200R, CD31, CD71, CLEC9A, CD1C for AXL/SIGLEC6;
(C) when the mRNA comprises a sequence encoding a CFP and the population of mRNA-electroporated cells are in the presence of cells expressing a cell-surface antigen to which the CFP specifically binds, the population of mRNA-electroporated cells one or more cytokines selected from the group consisting of IL-1beta, TNFalpha, IFN-alpha, MIP-1alpha, IL-8, eotaxin, PIGF-1, CCL3, IL6 and IL10; and
(D) when the mRNA comprises a sequence encoding a CFP and the population of mRNA-electroporated cells are in the presence of GMCSF, IL-4 IL-10, TGFbeta TCM and/or MCSF, the expression of MHCI or MHCII is upregulated in the population of mRNA-electroporated cells to a greater extent compared to a corresponding population of cells that have not been electroporated with the mRNA; and
(b) a pharmaceutically acceptable excipient; and
wherein when the mRNA comprises a sequence encoding a CFP, the population of mRNA-electroporated cells in the pharmaceutical composition do not exhibit tonic signaling through the CFP ex vivo.