ANTI-TCR ANTIBODY MOLECULES AND USES THEREOF
The disclosure provides antibody molecules that bind to TCR Vβ regions and multispecific molecules comprising said antibody molecules. Additionally, disclosed are nucleic acids encoding the same, methods of producing the aforesaid molecules, pharmaceutical compositions comprising aforesaid molecules, and methods of treating a cancer using the aforesaid molecules.
1 . A method of clonal expansion of T cells that express a T cell receptor beta variable region (TCRβV) in a T cell population, the method comprising:
contacting the T cell population with a composition comprising a multispecific molecule,
wherein the multispecific molecule comprises a first domain that binds to a first target molecule and a second domain that binds to a second target molecule,
wherein the first target molecule is a TCRβV,
wherein the multispecific molecule is a TCRβV agonist,
wherein the second domain comprises a tumor-targeting domain, a cytokine molecule, or a stromal modifying domain, and
wherein the first domain contacts the TCRβV of a T cell receptor (TCR) expressed by the T cells in the T cell population, thereby clonally expanding the T cells in the T cell population that express the TCR comprising the TCRβV to form a clonally expanded T cell population, wherein the clonally expanded T cell population produces:
(a) a higher level of IL-2 and/or one or more IL2 receptors (IL-2Rs) compared to the level of the IL-2 and/or the one or more IL-2Rs produced by a non-clonally expanded T cell population produced by contacting the T cells with an anti-CD3 antibody at the same concentration, and/or
(b) a lower level of one or more type 1 pro-inflammatory cytokines according to a cytokine release assay compared to the level of the one or more type 1 pro-inflammatory cytokines produced by a non-clonally expanded T cell population produced by contacting the T cells with an anti-CD3 antibody at the same concentration according to the cytokine release assay.
2 . The method of claim 1 , wherein the clonally expanded T cell population produces a higher level of IL-2 compared to the level of the IL-2 produced by a non-clonally expanded T cell population produced by contacting T cells with the anti-CD3 antibody at the same concentration.
3 . The method of claim 1 , wherein the clonally expanded T cell population produces a higher level of CD25 compared to the level of CD25 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration.
4 . The method of claim 1 , wherein the clonally expanded T cell population produces a higher level of CD122 compared to the level of CD122 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration.
5 . The method of claim 1 , wherein the clonally expanded T cell population produces a lower level of IL-6 according to a cytokine release assay compared to the level of IL-6 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration according to the cytokine release assay.
6 . The method of claim 1 , wherein the clonally expanded T cell population produces a lower level of IL-1 beta according to a cytokine release assay compared to the level of IL-1 beta produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration according to the cytokine release assay.
7 . The method of claim 1 , wherein the clonally expanded T cell population produces a lower level of TNF alpha according to a cytokine release assay compared to the level of TNF alpha produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration according to the cytokine release assay.
8 . The method of claim 1 , wherein the clonally expanded T cell population produces
(a) a higher level of IL-2 compared to the level of the IL-2 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration;
(b) a higher level of CD25 compared to the level of CD25 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration;
(c) a higher level of CD122 compared to the level of CD122 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration;
(d) a lower level of IL-6 according to a cytokine release assay compared to the level of IL-6 produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration according to the cytokine release assay;
(e) a lower level of IL-1 beta according to a cytokine release assay compared to the level of IL-1 beta produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration according to the cytokine release assay; and
(f) a lower level of TNF alpha according to a cytokine release assay compared to the level of TNF alpha produced by a non-clonally expanded T cell population produced by contacting the T cells with the anti-CD3 antibody at the same concentration according to the cytokine release assay.
9 . The method of claim 1 , wherein the second domain binds to a second target molecule expressed by the same T cell expressing the TCRβV to which the first domain of the multispecific molecule binds.
10 . The method of claim 1 , wherein the multispecific molecule comprises at least two non-contiguous polypeptide chains,
wherein a first polypeptide chain of the at least two non-contiguous polypeptide chains comprises a first member of a dimerization module, and a second polypeptide chain of the at least two non-contiguous polypeptide chains comprises a second member of the dimerization module, and wherein the first polypeptide chain and the second polypeptide chain form a complex via the first member of the dimerization module and the second member of the dimerization module.
11 . The method of claim 10 , wherein the first polypeptide chain comprises the first domain and the second polypeptide chain comprises the second domain, and wherein:
(i) the first polypeptide chain comprises the first domain linked to the first member of the dimerization module, and the second polypeptide chain comprises the second domain linked to the second member of the dimerization module;
(ii) the first polypeptide chain comprises a first portion of the first domain linked to the first member of the dimerization module, and the second polypeptide chain comprises a first portion of the second domain linked to the second member of the dimerization module; wherein the at least two non-contiguous polypeptide chains comprises a third polypeptide chain comprising a second portion of the first domain and a fourth polypeptide chain comprising a second portion of the second domain;
(iii) the first polypeptide chain comprises a first portion of the first domain linked to the first member of the dimerization module, and the second polypeptide chain comprises the second domain linked to the second member of the dimerization module; wherein the at least two non-contiguous polypeptide chains comprises a third polypeptide chain comprising a second portion of the first domain; or
(iv) the first polypeptide chain comprises the first domain linked to the first member of the dimerization module, and the second polypeptide chain comprises a first portion of the second domain linked to the second member of the dimerization module; wherein the at least two non-contiguous polypeptide chains comprises a third polypeptide chain comprising a second portion of the second domain.
12 . The method of claim 11 , wherein the multispecific molecule further comprises a linker between the first domain and the first member of the dimerization module, a linker between the second domain and the second member of the dimerization module, a linker between the first portion of the first domain and the first member of the dimerization module, a linker between the first portion of the second domain and the second member of the dimerization module, or a combination thereof; and wherein the linker is selected from an IgG hinge, a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker; a rigid linker, a helical linker, and a non-helical linker.
13 . The method of claim 10 , wherein the first polypeptide chain comprises (a) the first domain or a first portion of the first domain and (b) the second domain or a first portion of the second domain, and wherein the first polypeptide chain comprises:
(i) the first domain linked to the first member of the dimerization module linked to the second domain;
(ii) a first portion of the first domain linked to the first member of the dimerization module linked to a first portion of the second domain, wherein the at least two non-contiguous polypeptide chains comprises a third polypeptide chain comprising a second portion of the first domain and a fourth polypeptide chain comprising a second portion of the second domain;
(iii) a first portion of the first domain linked to the first member of the dimerization module linked to the second domain, wherein the at least two non-contiguous polypeptide chains comprises a third polypeptide chain comprising a second portion of the first domain; or
(iv) the first domain linked to the first member of the dimerization module linked to a first portion of the second domain, wherein the at least two non-contiguous polypeptide chains comprises a third polypeptide chain comprising a second portion of the second domain.
14 . The method of claim 13 , wherein the multispecific molecule further comprises a linker between the first domain and the first member of the dimerization module, a linker between the first portion of the first domain and the first member of the dimerization module, a linker between the first member of the dimerization module and the second domain, a linker between the first member of the dimerization module and the first portion of the second domain, or a combination thereof; and wherein the linker is selected from an IgG hinge, a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, and a non-helical linker.
15 . The method of claim 1 , wherein the multi specific molecule comprises a polypeptide sequence comprising:
(i) the first domain linked to the second domain;
(ii) a first portion of the first domain linked to a first portion of the second domain, wherein the polypeptide sequence further comprises a second portion of the first domain and a second portion of the second domain;
(iii) a first portion of the first domain linked to the second domain, wherein the polypeptide sequence further comprises a second portion of the first domain; or
(iv) the first domain linked to a first portion of the second domain, wherein the polypeptide sequence further comprises a second portion of the second domain.
16 . The method of claim 15 , wherein the polypeptide sequence further comprises a linker between the first domain and the second domain, a linker between the first portion of the first domain and the first portion of the second domain, a linker between the first portion of the first domain and the second domain, a linker between the first domain and the first portion of the second domain, or a combination thereof, and
wherein the linker is selected from an IgG hinge, a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, and a non-helical linker.
17 . The method of claim 1 , wherein the TCRβV is TCRβV1, TCRβV2, TCRβV3, TCRβV4, TCRβV5, TCRβV6, TCRβV7, TCRβV8, TCRβV9, TCRβV10, TCRβV11, TCRβV12, TCRβV19, TCRβV20, TCRβV21, TCRβV23, TCRβV24, TCRβV25, TCRβV26, TCRβV27, TCRβV28, TCRβV29 or TCRβV30.
18 . The method of claim 1 , wherein the TCRβV is TCRβV2, TCRβV4-1, TCRβV4-2, TCRβV5-1, TCRβV5-5, TCRβV5-6, TCRβV6, TCRβ6-5, TCRβV6-6, TCRβV6-9, TCRβV7-2, TCRβV7-3, TCRβV7-8, TCRβV7-9, TCRβV9, TCRβV10-1, TCRβV10-2, TCRβV10-3, TCRβV11-2, TCRβV12-3, TCRβV12-4, TCRβV12-5, TCRβV19, TCRβV20-1, TCRβV21, TCRβV24-1, TCRβV25-1 or TCRβV28.
19 . The method of claim 1 , wherein the TCRβV is TCRβV2, TCRβV3-1, TCRβV4-1, TCRβV4-2, TCRβV5-1, TCRβV5-4, TCRβV5-5, TCRβV5-6, TCRβV6-1, TCRβV6-5, TCRβV6-6, TCRβV7-3, TCRβV7-6, TCRβV7-8, TCRβV9, TCRβV11-2, TCRβV19, TCRβV20-1, TCRβV24-1, TCRβV27, TCRβV28, TCRβV29-1 or TCRβV30.
20 . The method of claim 1 , wherein second target molecule is selected from the group consisting of BCMA, FcRH5, CD19, CD20, CD22, CD30, CD33, CD38, CD47, CD99, CD123, CLEC12, CD179A, SLAMF7, PDL1, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PSMA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2/neu, Telomerase, SAP-1, Survivin, NY-ESO-1/LAGE-1, PRAME, SSX-2, Melan-A/MART-1, gp100/pmell7, Tyrosinase, MC1R, b-catenin, BRCA1/2, CDK4, CML66, Fibronectin, p53, Ras, TGF-B receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, a actinin-4, TRP1/gp75, TRP2, gangliosides, WT1, Epidermal growth factor receptor (EGFR), MART-2, MUC1, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrin, a carbohydrate, IGF1R, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.
21 . The method of claim 1 , wherein the second domain is an NK cell engager, a T cell engager, a B cell engager, a dendritic cell engager, or a macrophage cell engager.
22 . The method of claim 21 , wherein the second domain is a T cell engager and wherein the second target molecule is a TCRβV other than the TCRβV to which the first domain binds.
23 . The method of claim 21 , wherein the second target molecule is not a TCRβV.
24 . The method of claim 21 , wherein the second target molecule is CD19, CD3 or CD123.
25 . The method of claim 1 , wherein the second domain comprises a tumor-targeting domain and the second target molecule is a cancer antigen.
26 . The method of claim 25 , wherein the cancer antigen is a hematological cancer antigen, a solid tumor antigen, a metastatic cancer antigen, a soft tissue tumor antigen, a cancer antigen of a metastatic lesion or a stromal antigen.
27 . The method of claim 26 , wherein the cancer antigen is:
(i) the solid tumor antigen, wherein the solid tumor is pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, or liver cancer; or
(ii) the hematological cancer antigen, wherein the hematological cancer is a B-cell malignancy or a T cell malignancy.
28 . The method of claim 1 , wherein the second domain comprises a cytokine molecule selected from the group consisting of interleukin-2 (IL-2) or a functional fragment or variant thereof, interleukin-7 (IL-7) or a functional fragment or variant thereof, interleukin-12 (IL-12) or a functional fragment or variant thereof, interleukin-15 (IL-15) or functional fragment or variant thereof, interleukin-18 (IL-18) or a functional fragment or variant thereof, interleukin-21 (IL-21) or a functional fragment or variant thereof, and interferon gamma or a functional fragment or variant thereof.
29 . The method of claim 1 , wherein contacting comprises contacting the T cell population with the composition comprising a multispecific molecule at a concentration of at least 0.01 nM.
30 . The method of claim 1 , wherein the T cell population is an ex vivo T cell population.