Granted Patent B2

US 12,496,274 · App. 17/293,830 · Granted Dec 16, 2025

Inventors: Geoffrey A. von Maltzahn (Somerville, MA); John Miles Milwid (Denver, CO); Jacob Rosenblum Rubens (Cambridge, MA); Michael Travis Mee (Montreal, CA); Neal Francis Gordon (Brookline, MA); Jagesh Vijaykumar Shah (Lexington, MA); Kyle Marvin Trudeau (Boston, MA); Brigham Jay Hartley (Long Island City, NY)

Assignee: FLAGSHIP PIONEERING INNOVATIONS V, INC.

A61K9/1272A61K38/164A61K38/45A61K38/465A61K47/549A61K48/00

Abstract

Claims (26)

1 . A fusosome comprising:

(a) a lipid bilayer comprising a plurality of lipids obtained from a source cell;

(b) a lumen surrounded by the lipid bilayer;

(c) a fusogen that is exogenous or overexpressed relative to the source cell, wherein the fusogen is disposed in the lipid bilayer, and wherein the fusogen comprises a paramyxoviral fusogen; and

(d) a nuclear payload agent for delivery to the nucleus of a target cell, wherein the nuclear payload agent comprises or encodes a protein having a nuclear localization signal (NLS).

2 . The fusosome of claim 1 , wherein the nuclear protein payload agent comprises a nuclear localization signal set forth in any one of SEQ ID NOS: 128-507 and 605-626.

3 . The fusosome of claim 1 , wherein:

the nuclear payload agent comprises or encodes: a nuclease, a transcription factor, a recombinase, an epigenetic factor, a post-transcriptional RNA modification factor, a non-coding RNA or ribonucleic protein, or a structural protein;

the nuclear payload agent comprises or encodes one or more of a transcriptional activator, transcriptional repressor, epigenetic modifier, histone acetyltransferase, histone deacetylase, histone methyltransferase, DNA methyltransferase, a DNA nickase, a site-specific DNA editing enzyme, optionally a deaminase, DNA transposase, DNA integrase, an RNA editor, an RNA splicing factor, or a PIWI protein; or

the nuclear payload agent comprises or encodes an antibody molecule, e.g., a Fab, an scFv, an scFab, a sdAb, a duobody, a minibody, a nanobody, a diabody, a zybody, a camelid antibody, a BiTE, a quadroma, or a bsDb.

4 . The fusosome of claim 1 , wherein the source cell is a primary cell, a cultured cell, an immortalized cell, or a cell line.

5 . The fusosome of claim 1 , wherein the source cell is allogeneic.

6 . The fusosome of claim 1 , wherein the source cell is autologous.

7 . The fusosome of claim 1 , wherein the fusosome is from a source cell having a modified genome that exhibits reduced immunogenicity.

8 . The fusosome of claim 1 , wherein the fusosome:

(i) has a diameter that is less than about 0.01% or 1% of that of the source cell;

(ii) is active at a pH of 6-8.

9 . The fusosome of claim 1 , wherein the fusosome comprises a targeting domain which localizes the fusosome to a target cell.

10 . The fusosome of claim 9 , wherein the targeting domain interacts with a target cell moiety on the target cell.

11 . The fusosome of claim 1 , wherein the target cell is in an organism or is a primary cell isolated from an organism.

12 . The fusosome of claim 1 , wherein the target cell is selected from an endothelial cell, a fibroblast, a blood cell, a stem cell, an embryonic stem cell, a myoblast, a parenchymal cell, an alveolar cell, a neuron, a precursor cell, a progenitor cell, or an immortalized cell.

13 . A pharmaceutical composition comprising the fusosome of claim 1 .

14 . The fusosome of claim 1 , wherein when contacted with a target cell population, the fusosome delivers the payload agent via a non-endocytic pathway.

15 . The fusosome of claim 14 , wherein the level of payload delivered via a non-endocytic pathway for a given fusosome composition is 0.1-0.95, 0.1-0.2, 0.2-0.3, 0.3-0.4, 0.4-0.5, 0.5-0.6, 0.6-0.7, 0.7-0.8, 0.8-0.9, 0.9-0.95, or at least 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater than a chloroquine treated reference cell.

16 . The fusosome of claim 1 , wherein the paramyxovirus fusogen comprises a Nipah virus protein F, a measles virus F protein, a tupaia paramyxovirus F protein, a paramyxovirus F protein, a Hendra virus F protein, a Henipavirus F protein, a Morbilivirus F protein, a respirovirus F protein, a Sendai virus F protein, a rubulavirus F protein, or an avulavirus F protein.

17 . The fusosome of claim 9 , wherein the fusogen comprises the targeting domain.

Assignments (4)

Continuity (2)

References Cited (400)