Compositions and methods to modify cells for therapeutic objectives

Patent metadata

Field	Value
Publication number	US-2016145348-A1
Application number	US-201414776661-A
Country	US
Kind code	A1
Filing date	Mar 14, 2014
Priority date	Mar 14, 2013
Publication date	May 26, 2016
Grant date	—

Abstract

The present disclosure provides compositions and methods that rapidly and selectively modify cells of the immune system to achieve therapeutic objectives. The methods can be practiced in vivo and any cell type that expresses a known marker can be targeted for a therapeutic objective. The present disclosure provides compositions and methods that rapidly and selectively modify cells of the immune system to achieve therapeutic objectives. The methods can be practiced in vivo and any cell type that expresses or is associated with a known marker can be targeted for a therapeutic objective by the modified cell.

First claim

1 - 42 . (canceled)

43 . A synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the nanoparticle; and (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the nanoparticle.

44 . A synthetic nanocarrier of claim 43 further comprising an endosomal release agent extending from the surface of the nanoparticle and (ii) a nuclear localization signal within the pores of the nanoparticle.

45 . A synthetic nanocarrier of claim 43 wherein the synthetic nanocarrier comprises a liposome, polymeric particle, metallic particle, polymeric micelle, polyethyleneimine (PEI)/DNA complex, or a combination thereof.

46 . A synthetic nanocarrier of claim 43 wherein the lymphocyte-directing agent selectively binds to lymphocytes in vivo.

47 . A synthetic nanocarrier of claim 43 wherein the lymphocyte-directing agent selectively binds T cells, NK cells, monocytes, macrophages, B cells, hematopoietic stem cells or a combination thereof.

48 . A synthetic nanocarrier of claim 43 wherein the lymphocyte-directing agent selectively binds CCR7; CD3; CD4; CD5; CD8; CD16; CD19; CD20; CD21; CD22;
CD25; CD28; CD35; CD40; CD45RA; CD45RO; CD52; CD62L; CD80; CD95; CD127;
or CD137.

49 . A synthetic nanocarrier of claim 43 wherein the lymphocyte-directing agent selectively binds CD8.

50 . A synthetic nanocarrier of claim 43 wherein the lymphocyte-directing agent comprises a binding domain selected from a lymphocyte receptor ligand, lymphocyte receptor antibody, lymphocyte receptor peptide aptamer, lymphocyte receptor nucleic acid aptamer, lymphocyte receptor spiegelmer or a combination thereof.

51 . A synthetic nanocarrier of claim 49 wherein the binding domain consists of an ScFv fragment of a T-cell a chain antibody; T-cell β chain antibody; T-cell γ chain antibody; T-cell Δ chain antibody; CCR7 antibody; CD3 antibody; CD4 antibody; CD5 antibody; CD7 antibody; CD8 antibody; CD11b antibody; CD11c antibody; CD16 antibody; CD19 antibody; CD20 antibody; CD21 antibody; CD22 antibody; CD25 antibody; CD28 antibody; CD34 antibody; CD35 antibody; CD40 antibody; CD45RA antibody; CD45RO antibody; CD52 antibody; CD56 antibody; CD62L antibody; CD68 antibody; CD80 antibody; CD95 antibody; CD117 antibody; CD127 antibody; CD133 antibody; CD137 (4-1BB) antibody; CD163 antibody; F4/80 antibody; IL-4Rα antibody;
Sca-1 antibody; CTLA-4 antibody; GITR antibody GARP antibody; LAP antibody;
granzyme B antibody; LFA-1 antibody; transferrin antibody; or perforin antibody.

52 . A synthetic nanocarrier of claim 43 wherein the polynucleotide is a plasmid, a minicircle plasmid, or an mRNA molecule.

53 . A synthetic nanocarrier of claim 52 wherein the polynucleotide is a minicircle plasmid encoding a hyperactive transposase.

54 . A synthetic nanocarrier of claim 43 comprising a S/MAR element, a PiggyBac transposase-containing plasmid, a Sleeping Beauty transposase-containing plasmid; a homo sapiens transposon-derived Buster1 transposase-like protein gene; a human endogenous retrovirus H protease/integrase-derived ORF1; a homo sapiens Cas-Br-M (murine) ecotropic retroviral transforming sequence; a homo sapiens endogenous retroviral sequence K; a homo sapiens endogenous retroviral family W sequence; a homo sapiens LINE-1 type transposase domain; or a homo sapiens pogo transposable element.

55 . A synthetic nanocarrier of claim 43 wherein the CAR targeting agent comprises a binding domain for a marker associated with an unwanted cell type.

56 . A synthetic nanocarrier of claim 55 wherein the unwanted cell type is a cancer cell.

57 . A synthetic nanocarrier of claim 43 wherein the CAR targeting agent is a surface antigen receptor or a receptor for an intracellular antigen presented by a Major Histocompatibility Complex antigen-presenting pathway.

58 . A synthetic nanocarrier of claim 44 wherein the endosomal release agent is selected from any one of SEQ ID NOs.29-50.

59 . A synthetic nanocarrier of claim 44 wherein the NLS is selected from any of SEQ ID NOs. 51-93.

60 . A method for treating a disease associated with an antigen, the method comprising: administering to a subject in need thereof, a synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle with an endosomal release agent extending from the surface of the nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the nanoparticle; (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the nanoparticle; and (iv) a nuclear localization signal within the pores of the nanoparticle thereby treating the disease in the subject.

61 . A method of selectively transfecting lymphocytes in a subject in vivo, the method comprising:
contacting lymphocytes within the subjection with a synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle with an endosomal release agent extending from the surface of the nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the nanoparticle; (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the nanoparticle; and (iv) a nuclear localization signal within the pores of the nanoparticle thereby transfecting lymphocytes in the subject in vivo.

Assignees

• Hutchinson Fred Cancer Res

Inventors

• Stephan Matthias

Classifications

• A61K9/5123

  Organic compounds, e.g. fats, sugars · CPC title

• A61K47/6849

  the antibody targeting a receptor, a cell surface antigen or a cell surface determinant · CPC title

• A61K38/1774

  Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules) · CPC title

• A61K9/1271 Primary

  Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) · CPC title

• C07K14/7051

  T-cell receptor (TcR)-CD3 complex · CPC title