US 2020/0369773 A1 · CELLULAR BASED THERAPIES TARGETING DISEASE-ASSOCIATED MOLECULAR MEDIATORS OF FIBROTIC, INFLAMMATORY AND AUTOIMMUNE CONDITIONS

Abstract

The invention provides chimeric antigen receptors (CARs), nucleic acid sequences encoding a CAR, vectors comprising a nucleic acid sequence encoding a CAR, cells expressing a CAR, pharmaceutical compositions comprising a cell expressing a CAR, wherein the CAR binds to a target molecule expressed on disease-associated macrophages or over- or aberrantly-expressed in fibrosis. The invention further provides vectors encoding a CAR and a fibrotic disease-modulatory molecule (FDMM), and cells expressing both a CAR and an FDMM. The invention also provides methods of treating a subject using a CAR, a nucleic acid sequence, a vector or vectors, or a CAR-expressing cell, a cell expressing both a CAR and an FDMM, or a pharmaceutical composition, and to methods of generating a CAR-expressing cell or a cell expressing both a CAR and an FDMM. The invention also provides methods of treating diseases, fibrotic conditions, inflammatory conditions, autoimmune conditions, and conditions associated with disease-associated macrophages (DAMs).

Claims16

Dependent claims

1-49. (canceled)

A chimeric antigen receptor (CAR) comprising:

(a) an antigen-binding (AB) domain that binds to a target molecule which is expressed on disease-associated macrophages (DAMs) in a patient or which is over- or aberrantly-expressed in fibrosis,
(b) a transmembrane (TM) domain,
(c) an intracellular signaling (ICS) domain,
(d) optionally a hinge that joins said AB domain and said TM domain, and
(e) optionally one or more costimulatory (CS) domains;
optionally wherein:
(i) said target molecule is selected from the group consisting of fibroblast growth factor-inducible 14 (Fn14), CD163, CD206, CD209, FIZZ2 CD11b, SR1, F4/80, LY6G, LY6C, CD68, CD115, MAC2, MARCO, CCL2, TNFAIP3, CD11c, CD16, CD14, CD64, CD32, CD36, CD169, CD204, IL-4R α, IL-13RA1, EDNRA, EDNRB, IL6R, PDGFRB, HMGCR, PDGFRA, KDR, FLT1, HLA-DQB1, FGFR3, FGFR1, FLT4, FGFR2, FGFR4, TGFBRI, TGFBRII, PTGIR, CD19, CD109, VDR, IL6, EPHA2, and FGR;
(ii) said AB domain comprises an antibody (Ab) or an antigen-binding fragment thereof that binds to said target molecule, wherein said Ab or antigen-binding fragment thereof is optionally selected from a group consisting of a monoclonal Ab, a monospecific Ab, a polyspecific Ab, a humanized Ab, a tetrameric Ab, a tetravalent Ab, a multispecific Ab, a single chain Ab, a domain-specific Ab, a single-domain Ab (dAb), a domain-deleted Ab, an scFc fusion protein, a chimeric Ab, a synthetic Ab, a recombinant Ab, a hybrid Ab, a mutated Ab, CDR-grafted Ab, a fragment antigen-binding (Fab), an F(ab′)2, an Fab′ fragment, a variable fragment (Fv), a single-chain Fv (scFv) fragment, an Fd fragment, a dAb fragment, a diabody, a nanobody, a bivalent nanobody, a shark variable IgNAR domain, a VHH Ab, a camelid Ab, and a minibody;
(iii) one or more domains of the CAR comprise the ligand TWEAK or an Fn14-binding portion thereof;
(iv) said AB domain comprises a nanobody having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (iv-a) the amino acid sequence of NbMMRm22.84 (SEQ ID NO:
110), (iv-b) the amino acid sequence encoded by SEQ ID NO:
210, (iv-c) the amino acid sequence of NbMMRm5.38 (SEQ ID NO:
114), or (iv-d) the amino acid sequence encoded by SEQ ID NO:
214;
(v) said AB domain competes for binding to CD206 with a nanobody having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (v-a) the amino acid sequence of NbMMRm22.84 (SEQ ID NO:
110), (v-b) the amino acid sequence encoded by SEQ ID NO:
210, (v-c) the amino acid sequence of NbMMRm5.38 (SEQ ID NO:
114), or (v-d) the amino acid sequence encoded by SEQ ID NO:
214;
(vi) said AB domain comprises an Ab or antigen-binding fragment thereof comprising the amino acid sequences of (a) the three CDRs of the nanobody NbMMRm22.84 (SEQ ID NOS:
111-113), or (b) the three CDRs of the nanobody NbMMRm5.38 (SEQ ID NOS:
115-117);
(vii) said AB domain comprises an Ab or antigen-binding fragment thereof comprising amino acid sequences at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to said CDR sequences;
(viii) said AB domain comprises (A) a variable heavy (VH) chain having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (A-a) to the amino acid sequence of the VH chain of AbP4A8 or AbP3G5 (SEQ ID NOS:
118 or 126, respectively), or (A-b) the amino acid sequence encoded by SEQ ID NO:
218 or 226;
(B) a variable light (VL) chain having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (B-a) to the amino acid sequence of the VL chain of AbP4A8 or AbP3G5 (SEQ ID NOS:
122 or 130, respectively), or (B-b) to the amino acid sequence encoded by SEQ ID NO:
222 or 230;
and (C) optionally, a linker that links said VH chain to said VL chain, wherein said linker optionally comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (C-a) SEQ ID NO:
140, or (C-b) the amino acid sequence encoded by SEQ ID NO:
240;
(ix) said AB domain comprises an scFv fragment comprising an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (ix-a) the amino acid sequence of scFvP4A8VHVL, scFvP4A8VLVH, scFvP3G5VHVL, or scFvP3G5VLVH (SEQ ID NOS:
141, 142, 143, or 144, respectively), or (ix-b) the amino acid sequence encoded by SEQ ID NOS:
241, 242, 243, or 244;
(x) said AB domain competes for binding to Fn14 with an scFv fragment comprising an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (x-a) to the amino acid sequence of scFvP4A8VHVL, scFvP4A8VLVH, scFvP3G5VHVL, or scFvP3G5VLVH (SEQ ID NOS:
141, 142, 143, or 144, respectively), or (x-b) the amino acid sequence encoded by SEQ ID NOS:
241, 242, 243, or 244;
(xi) said AB domain comprises an Ab or antigen-binding fragment thereof comprising the amino acid sequences of (xi-a) the three heavy chain CDRs (SEQ ID NOS:
119-121) and the three light chain CDRs (SEQ ID NOS:
123-125) of AbP4A8, or (xi-b) the three heavy chain CDRs (SEQ ID NOS:
127-129) and the three light chain CDRs (SEQ ID NOS:
131-133) of AbP3G5;
(xii) said AB domain comprises an Ab or antigen-binding fragment thereof comprising amino acid sequences at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to said CDR sequences;
(xiii) said AB domain and/or TM domain comprises TWEAK or the AB or TM portion thereof, optionally comprising an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (xiii-a) to the amino acid sequence of human TWEAK or mouse TWEAK (SEQ ID NO:
134, or 135, respectively), or to the AB or TM portion thereof, or (xiii-b) the amino acid sequence encoded by SEQ ID NO:
234, or 235;
(xiv) said TM domain is derived from the TM region, or a membrane-spanning portion thereof, of a protein selected from the group consisting of CD28, CD3 ε, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD45, CD64, CD80, CD86, CD134, CD137, CD154, TCR α, TCR β, and CD3 ζ;
(xv) said TM domain is derived from the TM region of CD28, or a membrane-spanning portion thereof, optionally comprising an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (xv-a) the amino acid sequence of human CD28 TM domain (SEQ ID NO:
146) or mouse CD28 TM domain (SEQ ID NO:
746), or a membrane-spanning portion of either domain, or (xv-b) the amino acid sequence encoded by SEQ ID NO:
246 or SEQ ID NO:
846;
(xvi) said ICS domain is derived from a cytoplasmic signaling sequence, or a functional fragment thereof, of a protein selected from the group consisting of CD3 ζ, a lymphocyte receptor chain, a TCR/CD3 complex protein, an Fc receptor (FcR) subunit, an IL-2 receptor subunit, FcR γ, FcR β, CD3 γ, CD3 δ, CD3 ε, CD5, CD22, CD66d, CD79a, CD79b, CD278 (ICOS), Fc ε RI, DAP10, and DAP12;
(xvii) said ICS domain is derived from a cytoplasmic signaling sequence of CD3 ζ, or a functional fragment thereof, said ICS domain optionally comprising an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (xvii-a) the amino acid sequence of human CD3 ζ ICS domain (SEQ ID NO:
147) or mouse CD3 ζ ICS domain (SEQ ID NO:
747), or a functional fragment of either domain, or (xvii-b) the amino acid sequence encoded by SEQ ID NO:
247 or SEQ ID NO:
847;
(xviii) said hinge is derived from CD28, said hinge optionally comprising an amino sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (xviii-a) the amino acid sequence of human CD28 hinge (SEQ ID NO:
145) or mouse CD28 hinge (SEQ ID:
745), or (xviii-b) the amino acid sequence encoded by SEQ ID NO:
245 or SEQ ID NO:
845;
(xix) at least one of said one or more CS domains is derived from a cytoplasmic signaling sequence, or functional fragment thereof, of a protein selected from the group consisting of CD28, DAP10, 4-1BB (CD137), CD2, CD4, CD5, CD7, CD8 α, CD8β, CD11a, CD11b, CD11c, CD11d, CD18, CD19, CD27, CD29, CD30, CD40, CD49d, CD49f, CD69, CD84, CD96 (Tactile), CD100 (SEMA4D), CD103, OX40 (CD134), SLAM (SLAMF1, CD150, IPO-3), CD160 (BY55), SELPLG (CD162), DNAM1 (CD226), Ly9 (CD229), SLAMF4 (CD244, 2B4), ICOS (CD278), B7-H3, BAFFR, BTLA, BLAME (SLAMF8), CEACAM1, CDS, CRTAM, GADS, GITR, HVEM (LIGHTER), IA4, ICAM-1, IL2R β, IL2R γ, IL7R α, ITGA4, ITGA6, ITGAD, ITGAE, ITGAL, ITGAM, ITGAX, ITGB1, ITGB2, ITGB7, KIRDS2, LAT, LFA-1, LIGHT, LTBR, NKG2C, NKG2D, NKp30, NKp44, NKp46, NKp80 (KLRF1), PAG/Cbp, PD-1, PSGL1, SLAMF6 (NTB-A, Ly108), SLAMF7, SLP-76, TNFR2, TRANCE/RANKL, VLA1, VLA-6, and CD83 ligand;
(xx) said CS domain is derived from a cytoplasmic signaling sequence of CD28, 4-1BB, or DAP10, or functional fragment thereof, said CS domain optionally comprising an amino sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to (xx-a) to the amino acid sequence of human CD28 CS domain, human 4-1BB CS domain, human DAP10 CS domain, or mouse CD28 CS domain (SEQ ID NO:
156, 157, 158, or 756, respectively), or (xx-b) the amino acid sequence encoded by SEQ ID NO:
256, 257, 258, or 856;
(xxi) (a) said AB domain comprises the amino acid sequence of NbMMRm22.84, NbMMRm5.38, scFvP4A8VHVL, scFvP4A8VLVH, scFvP3G5VHVL, or scFvP3G5VLVH (SEQ ID NOS:
110, 114, 141, 142, 143, or 144, respectively), or the antigen-binding portion of TWEAK,
(b) said TM domain is derived from the TM region of CD28 or the TM region of TWEAK, optionally comprising the amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to the amino acid sequence of human CD28 TM domain (SEQ ID NO:
146) or of mouse CD28TM domain (SEQ ID NO:
746), or a membrane-spanning portion of any of the foregoing TM domains, and
(c) said ICS domain is derived from a cytoplasmic signaling sequence of CD3ζ, optionally comprising the amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to the amino acid sequence of human CD3 ζ ICS domain (SEQ ID NO:
147) or of mouse CD3 ζ ICS domain (SEQ ID NO:
747) or a functional fragment of any of the foregoing ICS domains;
(xxii) said CAR comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% at least 99%, or 100% identical to
(a) NbMMRm22.84-CD28H-CD28TM-CD28CS-CD3zICS (SEQ ID NO:
160),
(b) NbMMRm5.38-CD28H-CD28TM-CD28CS-CD3zICS (SEQ ID NO:
161),
(c) scFvP4A8VHVL-CD28H-CD28TM-CD28CS-CD3zICS (SEQ ID NO:
162),
(d) scFvP4A8VLVH-CD28H-CD28TM-CD28CS-CD3zICS (SEQ ID NO:
163),
(e) scFvP3G5VHVL-CD28H-CD28TM-CD28CS-CD3zICS (SEQ ID NO:
164),
(f) scFvP3G5VLVH-CD28H-CD28TM-CD28CS-CD3zICS (SEQ ID NO:
165),
(g) CD3zICS-CD28CS-TWEAK (SEQ ID NO:
136)
(h) NbMMRm22.84-CD28H-CD28TM-41BBCS-CD3zICS (SEQ ID NO:
166),
(i) NbMMRm5.38-CD28H-CD28TM-41BBCS-CD3zICS (SEQ ID NO:
167),
(j) scFvP4A8VHVL-CD28H-CD28TM-41BBCS-CD3zICS (SEQ ID NO:
168),
(k) scFvP4A8VLVH-CD28H-CD28TM-41BBCS-CD3zICS (SEQ ID NO:
169),
(l) scFvP3G5VHVL-CD28H-CD28TM-41BBCS-CD3zICS (SEQ ID NO:
170),
(m) scFvP3G5VLVH-CD28H-CD28TM-41BBCS-CD3zICS (SEQ ID NO:
171),
(n) CD3zICS-41BBCS-TWEAK (SEQ ID NO:
137)
(o) NbMMRm22.84-CD28H-CD28TM-DAP10CS-CD3zICS (SEQ ID NO:
172),
(p) NbMMRm5.38-CD28H-CD28TM-DAP10CS-CD3zICS (SEQ ID NO:
173),
(q) scFvP4A8VHVL-CD28H-CD28TM-DAP10CS-CD3zICS (SEQ ID NO:
174),
(r) scFvP4A8VLVH-CD28H-CD28TM-DAP10CS-CD3zICS (SEQ ID NO:
175),
(s) scFvP3G5VHVL-CD28H-CD28TM-DAP10CS-CD3zICS (SEQ ID NO:
176),
(t) scFvP3G5VLVH-CD28H-CD28TM-DAP10CS-CD3zICS (SEQ ID NO:
177),
(u) CD3zICS-DAP10CS-TWEAK (SEQ ID NO:
138)
(v) NbMMRm22.84-mCD28H-mCD28TM-mCD28CS-mCD3zICS (SEQ ID NO:
760),
(w) NbMMRm5.38-mCD28H-mCD28TM-mCD28CS-mCD3zICS (SEQ ID NO:
761),
(x) scFvP4A8VHVL-mCD28H-mCD28TM-mCD28CS-mCD3zICS (SEQ ID NO:
762),
(y) scFvP4A8VLVH-mCD28H-mCD28TM-mCD28CS-mCD3zICS (SEQ ID NO:
763),
(z) scFvP3G5VHVL-mCD28H-mCD28TM-mCD28CS-mCD3zICS (SEQ ID NO:
764),
(aa) scFvP3G5VLVH-mCD28H-mCD28TM-mCD28CS-mCD3zICS (SEQ ID NO:
765),
(bb) mCD3zICS-mCD28CS-mTWEAK (SEQ ID NO:
766);
or
(cc) the amino acid sequence encoded by SEQ ID NO:
260, 261, 262, 263, 264, 265, 236, 266, 267, 268, 269, 270, 271, 237, 272, 273, 274, 275, 276, 277, 238, or 860, 861, 862, 863, 864, 865, or 866;
or
(xxiii) said CAR further comprises a cytotoxic agent conjugated to said AB domain.

135 more elements

Dependencies are inferred from the claim text.

Facts

Application

16/639,836Filed 20 Aug 2018

Priority

18 Aug 2017

Published

26 Nov 2020

Term ends

Not estimatedThe source records don’t support an estimate.

Classification18 codes

C07K 16/2851
Against receptors, cell surface antigens or cell surface determinants: against the lectin superfamily, e.g. CD23, CD72
A61K 40/11
Characterised by the cell type used: t-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer…
A61K 40/22
Characterised by the effect or the function of the cells: immunosuppressive or immunotolerising
A61K 40/31
Characterised by the recombinant expression of specific molecules in the cells of the immune system: chimeric antigen receptors [CAR]

Cellular based therapies targeting disease-associated molecular mediators of fibrotic, inflammatory and autoimmune conditions

Life of this patent

Cellular based therapies targeting disease-associated molecular mediators of fibrotic, inflammatory and autoimmune conditions

Life of this patent

Abstract

Claims16