MND promoter chimeric antigen receptors

• US 10,774,343 B2
• Filed: 04/24/2015
• Issued: 09/15/2020
• Est. Priority Date: 04/25/2014
• Status: Active Grant

• Current Assignee: 2seventy Bio, Inc. (bluebird bio, Inc.)
• Original Assignee: bluebird bio, Inc.
• Inventors: Morgan, Richard, Friedman, Kevin, Ryu, Byoung
• Primary Examiner(s): Bristol, Lynn A
• Application Number:
  US15/306,724
• Publication Number:
  US 20170051308A1
• Time in Patent Office: 1,971 Days
• Field of Search: 4242771
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/001104 Epidermal growth factor receptors [EGFR]
• CPC Class Codes: A61K 39/001106 Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ErbB4
• CPC Class Codes: A61K 39/001109 Vascular endothelial growth factor receptors [VEGFR]
• CPC Class Codes: A61K 39/001112 CD19 or B4
• CPC Class Codes: A61K 39/001113 CD22, BL-CAM, siglec-2 or sialic acid- binding Ig-related lectin 2
• CPC Class Codes: A61K 39/001119 Receptors for interleukins [IL]
• CPC Class Codes: A61K 39/001122 Ephrin Receptors [Eph]
• CPC Class Codes: A61K 39/001124 CD20
• CPC Class Codes: A61K 39/001128 CD44 not IgG
• CPC Class Codes: A61K 39/001129 Molecules with a "CD" designation not provided for elsewhere
• CPC Class Codes: A61K 39/001138 Tumor necrosis factors [TNF] or CD70
• CPC Class Codes: A61K 39/00115 Apoptosis related proteins, e.g. survivin or livin
• CPC Class Codes: A61K 39/001166 Adhesion molecules, e.g. NRCAM, EpCAM or cadherins
• CPC Class Codes: A61K 39/001168 Mesothelin [MSLN]
• CPC Class Codes: A61K 39/00117 Mucins, e.g. MUC-1
• CPC Class Codes: A61K 39/001171 Gangliosides, e.g. GM2, GD2 or GD3
• CPC Class Codes: A61K 39/001174 Proteoglycans, e.g. glypican, brevican or CSPG4
• CPC Class Codes: A61K 39/00118 : from embryonic or fetal origin
• CPC Class Codes: A61K 39/001182 : Carcinoembryonic antigen [CEA]
• CPC Class Codes: A61K 39/001184 : Cancer testis antigens, e.g. SSX, BAGE, GAGE or SAGE
• CPC Class Codes: A61K 39/001186 : MAGE
• CPC Class Codes: A61K 39/001188 : NY-ESO
• CPC Class Codes: A61K 39/001189 : PRAME
• CPC Class Codes: A61K 39/001193 : Prostate associated antigens e.g. Prostate stem cell antigen [PSCA]; Prostate carcinoma tumor antigen [PCTA]; PAP or PSGR
• CPC Class Codes: A61K 39/001195 : Prostate specific membrane antigen [PSMA]
• 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 39/464412 : CD19 or B4
• CPC Class Codes: A61K 39/464417 : Receptors for tumor necrosis factors [TNF], e.g. lymphotoxin receptor [LTR], CD30
• 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: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/7051 : T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70517 : CD8
• CPC Class Codes: C07K 14/70578 : NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 16/2803 : against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/30 : from tumour cells
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/02 : containing a signal sequence
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C12N 15/86 : Viral vectors
• CPC Class Codes: C12N 2740/15043 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

Vector compositions comprising a myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted (MND) promoter operably linked to a chimeric antigen receptor (CAR) are provided.

Claims

1. A lentiviral vector comprising a polynucleotide comprising a myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted (MND) promoter operably linked to a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR comprises:

(a) a signal peptide;

(b) an scFv that binds B cell maturation antigen (BCMA);

(c) a CD8α

hinge domain as set forth in SEQ ID NO;

28;

(d) a CD8α

transmembrane domain;

(e) a CD137 co-stimulatory signaling domain; and

a CD3ζ

primary signaling domain.

2. The lentiviral vector of claim 1, wherein lentiviral vector selected from the group consisting of:

human immunodeficiency virus (HIV);

visna-maedi virus (VMV) virus;

caprine arthritis-encephalitis virus (CAEV);

equine infectious anemia virus (EIAV);

feline immunodeficiency virus (FIV);

bovine immune deficiency virus (BIV); and

simian immunodeficiency virus (SIV).

3. The lentiviral vector of claim 1, comprising a left (5′

) retroviral LTR, a Psi (Ψ

) packaging signal, a central polypurine tract/DNA flap (cPPT/FLAP), a retroviral export element;

the MND promoter operably linked to the CAR of claim 1; and

a right (3′

) retroviral LTR.

4. The lentiviral vector of claim 3, further comprising:

a) a heterologous polyadenylation sequence;

b) a heterologous polyadenylation sequence that is a bovine growth hormone polyadenylation or signal rabbit β

-globin polyadenylation sequence;

or

c) a hepatitis B virus posttranscriptional regulatory element (HPRE) or woodchuck post-transcriptional regulatory element (WPRE).

5. The lentiviral vector of claim 3, wherein:

a) the promoter of the 5′

LTR is replaced with a heterologous promoter;

b) the promoter of the 5′

LTR is replaced with a heterologous promoter selected from the group consisting of;

a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or a Simian Virus 40 (SV40) promoter.

6. The lentiviral vector of claim 3, wherein:

a) the 5′

LTR or 3′

LTR is a lentivirus LTR;

b) the 3′

LTR comprises one or more modifications;

c) the 3′

LTR comprises one or more deletions;

d) the 3′

LTR is a self-inactivating (SIN) LTR.

7. An isolated immune effector cell comprising the lentiviral vector of claim 1.

8. The immune effector cell of claim 7, wherein the immune effector cell is a T lymphocyte.

9. A composition comprising the immune effector cell of claim 7 and a physiologically acceptable excipient.

10. A composition comprising the immune effector cell of claim 8 and a physiologically acceptable excipient.

11. A method of making an immune effector cell comprising isolating CD34+ cells from bone marrow, cord blood or mobilized peripheral blood from a subject, and introducing the lentiviral vector of claim 1 into the isolated CD34+ cells.