US 11,903,974 B2
Methods and compositions relating to chondrisomes from cultured cells
Geoffrey A. von Maltzahn, Somerville, MA (US); John Miles Milwid, Denver, CO (US); Michael Travis Mee, Montreal (CA); Jacob Rosenblum Rubens, Cambridge, MA (US); David Chess, Waltham, MA (US); Kyle Marvin Trudeau, Boston, MA (US); Kiana Mahdaviani, Chestnut Hill, MA (US); Jacob Feala, Franklin, MA (US); James D. McCully, Marblehead, MA (US); and Douglas B. Cowan, Brighton, MA (US)
Assigned to FLAGSHIP PIONEERING INNOVATIONS V, INC., Cambridge, MA (US); and The Children's Medical Center Corporation, Boston, MA (US)
Appl. No. 15/779,736
Filed by FLAGSHIP PIONEERING INNOVATIONS V, INC., Cambridge, MA (US); and The Children's Medical Center Corporation, Boston, MA (US)
PCT Filed Nov. 30, 2016, PCT No. PCT/US2016/064238
§ 371(c)(1), (2) Date May 29, 2018,
PCT Pub. No. WO2017/095940, PCT Pub. Date Jun. 8, 2017.
Claims priority of provisional application 62/261,169, filed on Nov. 30, 2015.
Claims priority of provisional application 62/261,157, filed on Nov. 30, 2015.
Claims priority of provisional application 62/261,170, filed on Nov. 30, 2015.
Prior Publication US 2020/0306315 A1, Oct. 1, 2020
Int. Cl. A61K 35/35 (2015.01); A61K 35/34 (2015.01); A61K 35/14 (2015.01); A61K 35/19 (2015.01); A61K 38/17 (2006.01); A61K 35/33 (2015.01); G01N 33/15 (2006.01); C12N 15/87 (2006.01); A61K 35/12 (2015.01); A61P 3/00 (2006.01); A61K 9/00 (2006.01)
CPC A61K 35/35 (2013.01) [A61K 35/12 (2013.01); A61K 35/14 (2013.01); A61K 35/19 (2013.01); A61K 35/33 (2013.01); A61K 35/34 (2013.01); A61K 38/1709 (2013.01); A61P 3/00 (2018.01); C12N 15/87 (2013.01); G01N 33/15 (2013.01); A61K 9/0029 (2013.01)] 27 Claims
 
1. A method of enhancing metabolic function of a target cell or tissue, comprising delivering to the target cell or tissue a pharmaceutical composition comprising isolated chondrisomes derived from cultured cells, wherein the pharmaceutical composition is produced by a method comprising:
(a) providing a cell culture;
(b) dissociating the cells of the cell culture to produce a subcellular composition, wherein the dissociating comprises applying to the cells of the cell culture a first shear force followed by a second, higher shear force;
(c) separating the subcellular composition into a cellular debris fraction and a chondrisome enriched fraction, wherein the separating comprises comprising carrying out a first centrifugation and a second centrifugation, a fluid fraction produced by the first centrifugation is subjected to the second centrifugation, the cellular debris fraction is a solid or pelleted fraction produced by the second centrifugation, and the chondrisome enriched fraction is a fluid fraction produced by the second centrifugation;
(d) separating the chondrisome enriched fraction produced by the second centrifugation into a fraction containing chondrisomes and a fraction substantially lacking chondrisomes, wherein separating comprises carrying out a third centrifugation and a fourth centrifugation, a solid or pelleted fraction produced by the third centrifugation is subjected to the fourth centrifugation, the fraction containing chondrisomes is a solid or pellet fraction produced by the fourth centrifugation, and the fraction lacking chondrisomes is a fluid fraction produced by the fourth centrifugation; and
(e) suspending the fraction containing chondrisomes in a solution, thereby preparing a chondrisome preparation;
wherein the composition is delivered to the target cell or tissue in-vivo in a human subject; and
wherein the composition: (i) decreases serum cholesterol levels and/or triglycerides in the subject, or (ii) increases cardiac function in the subject.