Multispecific and multifunctional molecules and uses thereof

Abstract

Multispecific molecules that include i) a tumor-targeting moiety; and one, two or all of: (ii) an immune cell engager (e.g., chosen from an NK cell engager, a T cell engager, a B cell engager, a dendritic cell engager, or a macrophage cell engager); (iii) a cytokine molecule; and/or (iv) a stromal modifying moiety are disclosed. Additionally disclosed are nucleic acids encoding the same, methods of producing the aforesaid molecules, and methods of treating a cancer using the aforesaid molecules.

We claim: 
     
         1 .- 192 . (canceled) 
     
     
         193 . A multifunctional polypeptide molecule comprising:
 (i) a first antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof; and   (ii) a second moiety selected from the group consisting of a tumor-targeting moiety, a cytokine molecule, a stromal modifying moiety, and a second antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof;   wherein the multifunctional polypeptide molecule comprises one or more second moieties;   wherein the multifunctional polypeptide molecule further comprises a dimerization module comprising a first immunoglobulin chain constant region and a second immunoglobulin chain constant region;   wherein the first antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof is linked to the first immunoglobulin chain constant region; and   wherein the second moiety is linked to the first immunoglobulin chain constant region, the second immunoglobulin chain constant region, or a combination thereof.   
     
     
         194 . The multifunctional polypeptide molecule of  claim 193 , wherein the first immunoglobulin chain constant region comprises the first fragment crystallizable region (Fc region) and the second first immunoglobulin chain constant region comprises the second fragment crystallizable region (Fc region). 
     
     
         195 . The multifunctional polypeptide molecule of  claim 194 , wherein dimerization of the first Fc region and the second Fc region is enhanced by providing an Fc interface of the first Fc region and the second Fc region with one or more of a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer:homomultimer forms relative to a non-engineered interface. 
     
     
         196 . The multifunctional polypeptide molecule of  claim 193 , wherein the cytokine molecule is selected from the group consisting of interleukin-2 (TL-2) or a functional variant thereof, interleukin-7 (TL-7) or a functional variant thereof, interleukin-12 (IL-12) or a functional variant thereof, interleukin-15 (IL-15) or a functional variant thereof, interleukin-18 (IL-18) or a functional variant thereof, interleukin-21 (IL-21) or a functional variant thereof, and interferon gamma or a functional variant thereof. 
     
     
         197 . The multifunctional polypeptide molecule of  claim 196 , wherein the cytokine molecule comprises a receptor dimerizing domain. 
     
     
         198 . The multifunctional polypeptide molecule of  claim 193 , wherein the stromal modifying moiety is selected from the group consisting of a hyaluronidase, a collagenase, a chondroitinase, and a matrix metalloproteinase. 
     
     
         199 . The multifunctional polypeptide molecule of  claim 193 , wherein the multifunctional polypeptide molecule further comprises:
 a linker between the first antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof and the first immunoglobulin chain constant region,   a linker between the second moiety and the first immunoglobulin chain constant region,   a linker between the second moiety and the second immunoglobulin chain constant region, or   any combination thereof.   
     
     
         200 . The multifunctional polypeptide molecule of  claim 193 , wherein the multifunctional polypeptide molecule further comprises a cytokine molecules, a stromal modifying moiety, or a combination thereof linked to the first antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof, the tumor-targeting moiety, the second antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof, or any combination thereof. 
     
     
         201 . The multifunctional polypeptide molecule of  claim 200 , wherein the cytokine molecule is selected from the group consisting of interleukin-2 (TL-2) or a functional variant thereof, interleukin-7 (TL-7) or a functional variant thereof, interleukin-12 (IL-12) or a functional variant thereof, interleukin-15 (IL-15) or a functional variant thereof, interleukin-18 (IL-18) or a functional variant thereof, interleukin-21 (IL-21) or a functional variant thereof, and interferon gamma or a functional variant thereof. 
     
     
         202 . The multifunctional polypeptide molecule of  claim 201 , wherein the cytokine molecule comprises a receptor dimerizing domain. 
     
     
         203 . The multifunctional polypeptide molecule of  claim 200 , wherein the stromal modifying moiety is selected from the group consisting of a hyaluronidase, a collagenase, a chondroitinase, and a matrix metalloproteinase. 
     
     
         204 . The multifunctional polypeptide molecule of  claim 200 , wherein the multifunctional polypeptide molecule further comprises:
 a linker between the cytokine molecules, the stromal modifying moiety, or the combination thereof and the first antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof,   a linker between the cytokine molecules, the stromal modifying moiety, or the combination thereof and the tumor-targeting moiety,   a linker between the cytokine molecules, the stromal modifying moiety, or the combination thereof and the second antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof, or   any combination thereof.   
     
     
         205 . The multifunctional polypeptide molecule of  claim 193 , wherein the first antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof, the tumor-targeting moiety, the second antibody molecule that binds to NKp30 or NKp46 or antigen binding fragment thereof, or any combination thereof is a full length antibody, a dAb (domain antibody), an Fab, an Fab′, an F(ab′) 2  fragment, a single chain variable fragment (scFv), or a single domain antibody. 
     
     
         206 . The multifunctional polypeptide molecule of  claim 193 , wherein the tumor-targeting moiety comprises an antibody molecule that binds to a cancer antigen or antigen binding fragment thereof, wherein the cancer antigen is:
 (i) present on a hematological cancer, a solid tumor, a metastatic cancer, soft tissue tumor, metastatic lesion, or any combination thereof;   (ii) a tumor antigen, a stromal antigen, or a hematological antigen; or   (iii) a tumor antigen or stromal antigen present on a fibrotic or desmoplastic solid tumor.   
     
     
         207 . The multifunctional polypeptide molecule of  claim 206 , wherein the cancer antigen is:
 (i) selected from the group consisting of PDL1, mesothelin, CD47, prostate stem cell antigen (PSCA), GD2, prostate specific membrane antigen (PSMA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2/neu, Telomerase, SAP-1, Survivin, Melan-A/MART-1, MART-2, Gp100/pmel17, Tyrosinase, MC1R, β-catenin, BRCA1/2, CDK4, CML66, Fibronectin, p53, Ras, TGF-B receptor, MAGE, AFP, ETA, CA-125, NY-ESO-1/LAGE-1, PRAME, SSX-2, BAGE, GAGE, CDC27, α actinin-4, TRP1/gp75, TRP2, a ganglioside, WT1, Epidermal growth factor receptor (EGFR), CD20, MUC1, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TSTA, Folate receptor alpha, BING-4, L1-CAM, CAIX, EGFRvIII, gpA33, GD3, GM2, VEGFR, an Integrin, a carbohydrate, IGF1R, TRAILR1, TRAILR2, and RANKL;   (ii) a stromal antigen selected from the group consisting of fibroblast activating protease (FAP), TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W;   (iii) a hematological antigen selected from the group consisting of CD19, CD33, CD47, CD123, CD20, CD99, CD30, BCMA, CD38, CD22, SLAMF7, and NY-ESO1; or   (iv) a solid tumor antigen selected from the group consisting of PDL1, Mesothelin, HER3, IGF-1R, GD2, PSMA, CEA, Ron Kinase, and c-Met.   
     
     
         208 . The multifunctional polypeptide molecule of  claim 193 , wherein the multifunctional polypeptide molecule further comprises an immune cell engager selected from the group consisting of a T cell engager, a B cell engager, a dendritic cell engager, a macrophage cell engager, and any combination thereof; and
 wherein the T cell engager binds to CD3, TCRα, TCRO, TCRγ, TCRζ, ICOS, CD28, CD27, HVEM, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, or CD226;   wherein the B cell engager is a CD40 ligand, an OX40 ligand, a CD70 ligand, an antibody molecule that binds to OX40, an antibody molecule that binds to CD40, or an antibody molecule that binds to CD70;   wherein the dendritic cell engager is a CD2 agonist, an antibody molecule that binds to OX40, an OX40 ligand, a 41BB agonist, an agonist of a Toll-like receptor, a CD47 agonist, or a STING agonist; or   wherein the macrophage cell engager is a CD2 agonist, a CD40 ligand, an OX40 ligand, an antibody molecule that binds to OX40, an antibody molecule that binds to CD40, an antibody molecule that binds to CD70, an agonist of a Toll-like receptor, CD47, or a STING agonist.   
     
     
         209 . A pharmaceutical composition comprising the multifunctional polypeptide molecule of  claim 193  and a pharmaceutically acceptable carrier, excipient, or stabilizer. 
     
     
         210 . A method of treating cancer in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of  claim 209 , wherein the administering is effective to treat the cancer in the subject. 
     
     
         211 . The method of  claim 210 , wherein the cancer is:
 (i) a solid tumor cancer or a metastatic lesion; or   (ii) a hematological cancer.   
     
     
         212 . The method of  claim 211 , wherein:
 (i) the solid tumor is selected from the group consisting of pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and any combination thereof; or   (ii) the hematological cancer is selected from the group consisting of Hodgkin's lymphoma, Non-Hodgkin's lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, and acute lymphocytic leukemia.