COMPOSITIONS AND METHODS TO MODIFY CELLS FOR THERAPEUTIC OBJECTIVES

• US 20190330373A1
• Filed: 07/12/2019
• Published: 10/31/2019
• Est. Priority Date: 03/14/2013
• Status: Active Application

• Current Assignee: Fred Hutchinson Cancer Center
• Original Assignee: Fred Hutchinson Cancer Research Center (Fred Hutchinson Cancer Research Center|Argus Genetics|Mars)
• Inventors: Stephan, Matthias
• Application Number:
  US16/510,646
• Publication Number:
  US 20190330373A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2239/58 Prostate
• CPC Class Codes: A61K 38/177 Receptors; Cell surface antigens; Cell surface determinants
• CPC Class Codes: A61K 38/1774 Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/4632 T-cell receptors [TCR]; antibody T-cell receptor constructs
• CPC Class Codes: A61K 39/464453 Wilms tumor 1 [WT1]
• CPC Class Codes: A61K 39/464495 Prostate specific membrane antigen [PSMA]
• CPC Class Codes: A61K 39/464838 Viral antigens
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 47/6455 Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6913 the liposome being modified on its surface by an antibody
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: A61K 9/5123 Organic compounds, e.g. fats, sugars
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 16/2815 : against CD8
• CPC Class Codes: C07K 16/3069 : Reproductive system, e.g. ovaria, uterus, testes, prostate
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/74 : containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C12N 15/88 : using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2810/859 : from immunoglobulins

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.

Claims

1. A synthetic nanocarrier comprising (i) a lipid-coated porous nanoparticle (ii) a lymphocyte-directing agent extending from the surface of the lipid-coated porous nanoparticle;

and (iii) a polynucleotide encoding a chimeric antigen receptor (CAR) targeting agent within the pores of the lipid-coated porous nanoparticle.

3. A synthetic nanocarrier of claim 1 wherein the CAR is P28z.

4. A synthetic nanocarrier of claim 1 wherein the lipid coating is a liposome, a lipid bilayer, or a polymeric micelle.

5. A synthetic nanocarrier of claim 1 wherein the synthetic nanocarriers comprise liposomes, polymeric particles, metallic particles, polymeric micelles, polyethyleneimine (PEI)/DNA complexes, or a combination thereof.

6. A synthetic nanocarrier of claim 1 wherein the lipid coating encapsulates the lipid-coated porous nanoparticle.

7. A synthetic nanocarrier of claim 1 wherein the lymphocyte-directing agent selectively binds to lymphocytes in vivo.

8. A synthetic nanocarrier of claim 1 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.

9. A synthetic nanocarrier of claim 1 wherein the lymphocyte-directing agent selectively binds T cells, NK cells, monocytes, macrophages, B cells, hematopoietic stem cells, or a combination thereof.

10. A synthetic nanocarrier of claim 1 wherein the lymphocyte-directing agent selectively binds T-cell receptor motifs;

T-cell α

chains;

T-cell β

chains;

T-cell γ

chains;

T-cell Δ

chains;

CCR7;

CD3;

CD4;

CD5;

CD7;

CD8;

CD11b;

CD11c;

CD16;

CD19;

CD20;

CD21;

CD22;

CD25;

CD28;

CD34;

CD35;

CD40;

CD45RA;

CD45RO;

CD52;

CD56;

CD62L;

CD68;

CD80;

CD95;

CD117;

CD127;

CD133;

CD137 (4-1BB);

CD163;

F4/80;

IL-4Rα

;

Sca-1;

CTLA-4;

GITR;

GARP;

LAP;

granzyme B;

LFA-1;

or transferrin receptor.

11. A synthetic nanocarrier of claim 1 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.

12. A synthetic nanocarrier of claim 1 wherein the lymphocyte-directing agent comprises a binding domain selected from a T-cell α

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;

or transferrin receptor antibody.

13. A synthetic nanocarrier of claim 12 wherein the binding domain consists of or consists essentially of an scFv fragment of a T-cell α

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;

or transferrin receptor antibody.

14. A synthetic nanocarrier of claim 13 wherein the binding domain consists of consists of or consists essentially of the scFv fragment (SEQ ID NO. 1) of the PSMA-specific chimeric antigen receptor (CAR), P28z.

15. A synthetic nanocarrier of claim 1 wherein the polynucleotide is a plasmid, a minicircle plasmid, or an mRNA molecule.

16. A synthetic nanocarrier of claim 1 wherein the CAR targeting agent comprises a binding domain for a marker associated with an unwanted cell type.

17. A synthetic nanocarrier of claim 16 wherein the unwanted cell type is a cancer cell.

18. A synthetic nanocarrier of claim 16 wherein the marker is a cancer antigen.

19. A synthetic nanocarrier of claim 16 wherein the marker is a cancer antigen selected from A33;

BAGE;

Bcl-2;

β

-catenin;

CA125;

CA19-9;

CD5;

CD19;

CD20;

CD21;

CD22;

CD33;

CD37;

CD45;

CD123;

CEA;

c-Met;

CS-1;

cyclin B1;

DAGE;

EBNA;

EGFR;

ephrinB2;

estrogen receptor;

FAP;

ferritin;

folate-binding protein;

GAGE;

G250;

GD-2;

GM2;

gp75, gp100 (Pmel

     17);

HER-2/neu;

HPV E6;

HPV E7;

Ki-67;

LRP;

mesothelin, p53, PRAME;

progesterone receptor;

PSA;

PSMA;

MAGE;

MART;

mesothelin;

MUC;

MUM-1-B;

myc;

NYESO-1;

ras;

RORI;

survivin;

tenascin;

TSTA tyrosinase;

VEGF;

or VVT1.

20. A synthetic nanocarrier of claim 16 wherein the marker is PSMA.

21. A synthetic nanocarrier of claim 1 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.

24. A synthetic nanocarrier of claim 1 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.

25. A composition comprising a synthetic nanocarrier of claim 1.

26. A method of treating a subject having a condition associated with a cell type comprising:

administering a therapeutically effective amount of a synthetic nanocarrier of claim 1 to the subject thereby treating the subject.

27. A method of treating a subject having a condition associated with an unwanted cell type comprising:

administering a therapeutically effective amount of a composition of claim 25 to the subject thereby treating the subject.

28. A method of claim 27 wherein the unwanted cell type is an unwanted cancer cell.

29. A method of claim 28 wherein the unwanted cancer cell is selected from an adrenal cancer cell, a bladder cancer cell, a blood cancer cell, a bone cancer cell, a brain cancer cell, a breast cancer cell, a carcinoma cell, a cervical cancer cell, a colon cancer cell, a colorectal cancer cell, a corpus uterine cancer cell, an ear, nose and throat (ENT) cancer cell, an endometrial cancer cell, an esophageal cancer cell, a gastrointestinal cancer cell, a head and neck cancer cell, a Hodgkin'"'"'s disease cell, an intestinal cancer cell, a kidney cancer cell, a larynx cancer cell, a leukemia cell, a liver cancer cell, a lymph node cancer cell, a lymphoma cell, a lung cancer cell, a melanoma cell, a mesothelioma cell, a myeloma cell, a nasopharynx cancer cell, a neuroblastoma cell, a non-Hodgkin'"'"'s lymphoma cell, an oral cancer cell, an ovarian cancer cell, a pancreatic cancer cell, a penile cancer cell, a pharynx cancer cell, a prostate cancer cell, a rectal cancer cell, a sarcoma cell, a seminoma cell, a skin cancer cell, a stomach cancer cell, a teratoma cell, a testicular cancer cell, a thyroid cancer cell, a uterine cancer cell, a vaginal cancer cell, or a vascular tumor cell.

30. A method of claim 26 wherein the administering results in expression of the polynucleotide selectively by lymphocytes within 10 days;

within 9 days;

within 8 days;

within 7 days;

within 6 days;

within 5 days;

within 4 days;

or within 3 days of administration.

2. A synthetic nanocarrier of embodiment 1 further comprising an endosomal release agent extending from the surface of the lipid-coated porous nanoparticle and (ii) a nuclear localization signal (NLS) within the pores of the lipid-coated porous nanoparticle.

22. A synthetic nanocarrier of claim 2 wherein the endosomal release agent is selected from any one of SEQ ID NOs. 29-50 or combinations thereof.

23. A synthetic nanocarrier of claim 2 wherein the NLS is selected from any one of SEQ ID NOS. 51-93 or combinations thereof.

31. A method for treating a disease associated with an antigen, the method comprising:

administering to a subject in need thereof, a composition comprising a therapeutically effective amount of nanocarriers including a polynucleotide having a sequence that encodes a receptor for the antigen, thereby treating the disease.

32. A method of claim 31 wherein after the administering the nanocarriers are selectively incorporated into lymphocytes in the subject such that the lymphocytes express the receptor and subsequently bind to the antigen on cells associated with the disease thereby killing the cells.

33. A method for treating a disease associated with an antigen, the method comprising:

obtaining lymphocytes from a subject in need thereof;

combining the lymphocytes with a composition comprising nanocarriers including a polynucleotide having a sequence that encodes a receptor for the antigen, wherein the nanocarriers are selectively incorporated into the lymphocytes such that the lymphocytes express the receptor; and

administering the lymphocytes expressing the receptor to the subject, thereby treating the disease.

34. A method of claim 33 wherein after the administering, the lymphocytes bind to the antigen on cells associated with the disease thereby killing the cells.

35. A method of selectively transfecting lymphocytes in vivo, the method comprising:

contacting lymphocytes with nanocarriers comprising a polynucleotide having a sequence that encodes a receptor for an antigen associated with a disease, wherein the nanocarriers are selectively incorporated into the lymphocyte to release the polynucleotide such that the lymphocyte expresses the receptor, thereby transfecting the lymphocyte.

36. A method of claim 35 wherein the antigen comprises a tumor antigen.

37. A method of claim 35 wherein the antigen comprises a viral antigen.

38. A method of claim 35 wherein the lymphocytes comprise T-cells, NK cells, macrophages, monocytes, B cells, hematopoietic stem cells, or a combination thereof.

39. A method of claim 38 wherein the lymphocytes comprise T-cells.

40. A method of claim 35 wherein the disease is a cancer.

41. A method of claim 40 wherein the cancer comprises a leukemia, a lymphoma, a carcinoma, a sarcoma, or a melanoma.

42. A method of a claim 40 wherein the disease is prostate cancer.