Twin Immune Cell Engager

Abstract

A Twin Immune Cell Engager (TWICE) is a kit or composition for treating cancer comprising a first component and second component. Each component comprises a targeting moiety that binds a tumor antigen expressed by the cancer or an antigen expressed by a non-cancer cell in the tumor microenvironment. In some embodiments, the first and second components each comprise an immune cell binding domain capable of immune cell binding activity when binding the immune cell binding domain in the other component, and a complementary binding domain capable of binding to a complementary antigen when binding the complementary binding domain in the other component. In some embodiments, the first and/or second components comprise a complementary functional domain with activity when targeted to the cancer cell or its microenvironment. The present TWICE complexes offer a unique ability to combine multiple functions into a two-component complex that becomes activated in the tumor microenvironment. The present complexes, thus, provide meaningful advantages in having a single approach to administering a two-component complex that is localized to the tumor microenvironment and has the ability to effect two different signals to benefit patients. This unique construct offers benefits that were not present in prior art constructs.

1 - 25 . (canceled) 
     
     
         26 . A kit or composition for treating cancer in a patient comprising:
 a. a first component comprising a targeted immune cell binding agent comprising:
 i. a first targeting moiety that binds a tumor antigen expressed by the cancer; 
 ii. a first immune cell binding domain capable of immune cell binding activity when binding a second immune cell binding domain, wherein the second immune cell binding domain is not part of the first component, and wherein the first immune cell binding domain is either a VH domain or VL domain; 
 iii. a first inert binding partner for the first immune cell binding domain, wherein the first inert binding partner binds to the first immune cell binding domain such that the first immune cell binding domain does not bind to the second immune cell binding domain unless the inert binding partner is removed, wherein if the first immune cell binding domain is a VH domain, the inert binding partner is a VL domain and if the first immune cell binding domain is a VL domain, the inert binding partner is a VH domain; 
 iv. a protease cleavage site separating the first immune cell binding domain and the first inert binding partner, wherein the protease cleavage site is capable of releasing the inert binding partner from the immune cell binding domain in the presence of a protease:
 (1) expressed by the cancer or a tumor microenvironment cell; or 
 (2) colocalized to the cancer by a targeting moiety that is an antibody or antigen binding fragment thereof that binds (a) a tumor antigen expressed by the cancer and that is the same or different from the first and/or second targeting moiety in the agent or (b) an antigen expressed by a cell in the tumor microenvironment; and 
 
 v. a first complementary functional domain capable of immune cell binding, and: 
   b. a second component comprising a targeted immune cell binding agent comprising:
 i. a second targeting moiety; 
 ii. a second immune cell binding domain; and 
 iii. optionally a second complementary functional domain capable of immune cell binding. 
   
     
     
         27 . The kit or composition of  claim 26 , wherein the second component comprises a complementary functional domain; optionally wherein the complementary functional domain of the first and/or second component comprises a ligand for a receptor; further optionally wherein the complementary functional domain comprises a latent form of a member of the TGF-beta family or a cytokine. 
     
     
         28 . The kit or composition of  claim 27 , wherein the cytokine is IL-2, IL-7, IL-12, IL-15, GM-CSF, IFN-α, IFN-γ, or a member of the TNF-superfamily. 
     
     
         29 . The kit or composition of  claim 27 , wherein the complementary functional domain of the first and/or second component comprises an attenuated cytokine, optionally wherein the attenuated cytokine is a variant of IL-2, IL-7, IL-12, IL-15, GM-CSF, IFN-α, IFN-γ, or a member of the TNF-superfamily. 
     
     
         30 . The kit or composition of  claim 26 , wherein the second targeting moiety binds a tumor antigen expressed by the cancer. 
     
     
         31 . The kit or composition of  claim 26 , wherein the second component further comprises a second inert binding partner for the second immune cell binding domain, wherein the second inert binding partner binds to the second immune cell binding domain such that the second immune cell binding domain does not bind to the first immune cell binding domain unless the inert binding partner is removed, wherein if the second immune cell binding domain is a VH domain, the inert binding partner is a VL domain and if the second immune cell binding domain is a VL domain, the inert binding partner is a VH domain; and further wherein
 a protease cleavage site separates the second immune cell binding domain and the second inert binding partner, wherein the protease cleavage site is capable of releasing the inert binding partner from the immune cell binding domain in the presence of a protease:
 a. expressed by the cancer; or 
 b. colocalized to the cancer by a targeting moiety that is an antibody or antigen binding fragment thereof that binds (a) a tumor antigen expressed by the cancer and that is the same or different from the first and/or second targeting moiety in the agent or (b) an antigen expressed by a cell in the tumor microenvironment. 
   
     
     
         32 . The kit or composition of  claim 26 , wherein the first immune cell binding domain and the second immune cell binding domain are, when bound to each other, capable of binding a T cell, a macrophage, or a natural killer cell. 
     
     
         33 . The kit or composition of  claim 32 , wherein the first immune cell binding domain and the second immune cell binding domain are, when bound to each other, capable of binding a T cell; optionally wherein the first immune cell binding domain and the second immune cell binding domain are, when bound to each other, capable of binding CD3, the T-cell receptor, programmed cell death protein 1 (PD-1), cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), T-cell immunoglobulin and mucin-domain 3 (TIM-3), lymphocyte-activation gene 3 (LAG-3), killer-cell immunoglobulin-like receptor (KIR), CD28, CD137, OX40, CD27, GITR (TNFRSF18), TIGIT, or inducible T-cell costimulatory (ICOS). 
     
     
         34 . The kit or composition of  claim 32 , wherein the first immune cell binding domain and the second immune cell binding domain are, when bound to each other, capable of binding a macrophage; optionally wherein the first immune cell binding domain and the second immune cell binding domain are, when bound to each other, capable of binding CSF1R. 
     
     
         35 . The kit or composition of  claim 32 , wherein the first immune cell binding domain and the second immune cell binding domain are, when bound to each other, capable of binding a natural killer cell; optionally wherein the first immune cell binding domain and the second immune binding domain are, when bound to each other, capable of binding CD16A. 
     
     
         36 . The kit or composition of  claim 26 , wherein the first and/or second targeting moiety comprises an antibody or antigen binding fragment thereof; optionally wherein the first and/or second targeting moiety comprises an antibody or antigen binding fragment thereof that binds α4-integrin; A33; ACVRL 1/ALK1; ADAM17; ALK; APRIL; BCMA; C242; CA125; Cadherin-19; CAIX; CanAg; Carbonic Anhydrase IX; CCN1; CCR4; CD123; CD133; CD137 (4-1BB); CD138/Syndecan1; CD19; CD2; CD20; CD22; CD30; CD33; CD37; CD38; CD4; CD40; CD44; CD45; CD48; CDS; CD52; CD56; CD59; CD70; CD70b; CD71; CD74; CD79b; CD80; CD86; CD98; CEA; CEACAM; CEACAM1; CK8; c-Kit; CLDN1; CLDN18; CLDN18.2; CLDN6; c-met/HGFR; c-RET; Cripto; CTLA-4; CXCR4; DKK-1; DLL3; DLL4; TRAIL-R2/DR5; DRS; EGFL7; EGFR; EGFRvIII; endoglin; ENPP3; EpCAM; EphA2; Episialin; FAP; FGFR1; FGFR2; FGFR3; FGFR4; fibronectin extra-domain B; FLT-3; flt4; folate receptor 1; GCC; GD2; GD3; Glypican-3; Glypicans; GM3; GPNMB; GPR49; GRP78; Her2/Neu; HER3/ERBB3; HLA-DR; ICAM-1; IGF-1R; IGFR; IL-3Ra; Integrin α5β1; Integrin α6β4; Integrin αV; Integrin αVβ3; Lewis Y; Lewis y/b antigen; LFL2; LIV-1; Ly6E; MCP-1; Mesothelin; MMP-9; MUC1; MUC18; MUC5A; MUC5AC; Myostatin; NaPi2b; Neuropilin 1; NGcGM3; NRP1; P-cadherin; PCLA; PD-1; PDGFRa; PD-L1; PD-L2; Phosphatidylserine; PIVKA-II; PLVAP; PRLR; Progastrin; PSCA; PSMA; RANKL; RG1; Siglec-15; SLAMF6; SLAMF7; SLC44A4, STEAP-1; TACSTD-2; Tenascin C; TPBG; TRAIL-R1/DR4; TROP-2; TWEAKR; TYRP1; VANGL2; VEGF; VEGF-C; VEGFR-2; or VEGF-R2. 
     
     
         37 . The kit or composition of  claim 36 , wherein the first and/or second targeting moiety comprises an antibody or antigen binding fragment thereof is an anti α4-integrin antibody; an anti-CD137 antibody; an anti-CCR4 antibody; an anti-CD123 antibody; an anti-CD133 antibody; an anti-CD138 antibody; an anti-CD19 antibody; an anti-CD20 antibody; an anti-CD22 antibody; an anti-CD33 antibody; an anti-CD38 antibody; an anti-CD40 antibody; an anti-CD49d antibody; an anti-CD52 antibody; an anti-CD70 antibody; an anti-CD74 antibody; an anti-CD79b antibody; an anti-CD80 antibody; an anti-CEA antibody; an anti-cMet antibody; an anti-Cripto antibody; an anti-CTLA-4 antibody; an anti-DLL3 antibody; an anti-TRAIL-2/DR5 antibody; an anti-E-cadherin antibody; an anti-endoglin antibody; an anti-EpCAM antibody; an anti-epidermal growth factor receptor antibody; an anti-FGFR3 antibody; an anti-fibronectin extra-domain B antibody; an anti-folate receptor 1 antibody; an anti-glypican 3 antibody; an anti-gp95/97 antibody; an anti-Her2 antibody; an anti-IGF-1R antibody; an anti-IL-13R antibody; an anti-IL-4 antibody; an anti-IL-6 antibody; an anti-MMP-9 antibody; an anti-MUC1 antibody; an anti-mucin core protein antibody; an anti-NGcGM3 antibody; an anti-P-cadherin antibody; an anti-PD-L1 antibody; an anti-p-glycoprotein antibody; an anti-PSCA antibody; an anti-PSMA antibody; an anti-SLAMF7 antibody; an anti-TRAIL-R1/DR4 antibody; an anti-transferrin antibody; an anti-TROP-2 antibody; or an anti-VEGF antibody. 
     
     
         38 . The kit or composition of  claim 36 , wherein the first and/or second targeting moiety comprises Alemtuzumab, Andecaliximab, Atezolizumab, Avelumab, BCD-100, Bevacizumab, BGB-A317, Blinatumomab, Brentuximab, BU59, Camrelizumab, Carotuximab, Catumaxomab, Cemiplimab, Cetuximab, Daratumumab, Depatuxizumab, Dinutuximab, DS-8201, Durvalumab, Edrecolomab, Elotuzumab, G544, Gemtuzumab, Glembatumumab, GP1.4, hp67.6, IBI308, Ibritumomab, Inotuzumab, Ipilimumab, Isatuximab, L19IL2, L19TNF, Margetuximab, Mirvetuximab, Mogamuizumab, Moxetumomab, Natalizumab, Necitumumab, Nivolumab, Obinutuzumab, Ofatumumab, Olaratumab, Oportuzumab, Panitumumab, PDR001, Pembrolizumab, Pertuzumab, Polatuzumab, Racotumomab, Ramucirumab, Rituximab, Rovalpituzumab, Sacituzumab, SM3, TAK-164, Tositumomab, Trastuzumab, Tremelimumab, Ublituximab, Urelumab, Utomilumab, XMAB-5574, or Zolbetuximab. 
     
     
         39 . The kit or composition of  claim 26 , wherein the first and/or second targeting moiety comprises a DNA aptamer, RNA aptamer, albumin, lipocalin, fibronectin, ankyrin, fynomer, Obody, DARPin, knotin, avimer, atrimer, anti-callin, affilin, affibody, bicyclic peptide, cys-knot, FN3 (adnectins, centryrins, pronectins, or TN3), or Kunitz domain. 
     
     
         40 . The kit or composition of  claim 26 , wherein the first and/or second targeting moiety comprises:
 a. IL-2, IL-4, IL-6, α-MSH, transferrin, folic acid, EGF, TGF, PD-1, IL-13, stem cell factor, insulin-like growth factor (IGF), or CD40;   b. a full-length sequence of IL-2, IL-4, IL-6, α-MSH, transferrin, folic acid, EGF, TGF, PD-1, IL-13, stem cell factor, insulin-like growth factor (IGF), or CD40; or   c. a truncated form, analog, variant, or derivative of IL-2, IL-4, IL-6, α-MSH, transferrin, folic acid, EGF, TGF, PD-1, IL-13, stem cell factor, insulin-like growth factor (IGF), or CD40.   
     
     
         41 . The kit or composition of  claim 26 , wherein the first and/or second targeting moiety binds the IL-2 receptor, IL-4, IL-6, melanocyte stimulating hormone receptor (MSH receptor), transferrin receptor (TR), folate receptor 1 (FOLR), folate hydroxylase (FOLH1), EGF receptor, PD-L1, PD-L2, IL-13R, CXCR4, IGFR, or CD40L. 
     
     
         42 . The kit or composition of  claim 26 , wherein the second targeting moiety binds an antigen expressed by a tumor microenvironment cell; optionally wherein the tumor microenvironment cell is a fibroblast or macrophage; further optionally wherein the tumor microenvironment cell is a fibroblast and the antigen expressed by the fibroblast is fibroblast activation protein. 
     
     
         43 . The kit or composition of  claim 42 , wherein the antigen expressed by a fibroblast is fibroblast activation protein or the antigen expressed by a macrophage is MAC-1/CD11b or sideroflexin 3. 
     
     
         44 . A method of delivering a cytokine to an immune cell of a patient comprising administering the composition of  claim 26  to the patient, wherein the first and/or second complementary functional domain of the composition comprise IL-2, IL-7, IL-12, IL-15, GM-CSF, IFN-α, IFN-γ, or a member of the TNF-superfamily.