Targeted chimeric proteins and uses thereof

Abstract

The present invention relates, in part, to chimeric proteins comprising at least one targeting moiety that recognizes and binds SIRP1α and their use as diagnostic and therapeutic agents. The present invention further relates to pharmaceutical compositions comprising the chimeric proteins and their use in the treatment of various diseases.

What is claimed is: 
     
         1 . A chimeric protein comprising:
 (a) a targeting moiety comprising a recognition domain which recognizes and binds to SIRP1α; and   (b) a modified signaling agent, said modified signaling agent having one or more mutations that confer improved safety relative to a wild type signaling agent as compared to a wild type signaling agent, and   wherein the targeting moiety and modified signaling agent are optionally connected with one or more linkers.   
     
     
         2 . The chimeric protein of  claim 1 , further comprising one or more additional targeting moieties. 
     
     
         3 . The chimeric protein of  claim 2 , wherein the one or more additional targeting moieties comprise a recognition domain that recognizes and binds an antigen or receptor on a tumor cell. 
     
     
         4 . The chimeric protein of  claim 2 , wherein the one or more additional targeting moieties comprise a recognition domain that recognizes and binds an antigen or receptor on an immune cell. 
     
     
         5 . The chimeric protein of  claim 4 , wherein the immune cell is selected from a macrophage, a monocyte, and a dendritic cell. 
     
     
         6 . The chimeric protein of any one of  claims 2 - 5 , wherein the one or more additional targeting moieties recognizes one or more of PD-L1, PD-L2, PD-1, and Clec9A. 
     
     
         7 . The chimeric protein of any one of the above claims, wherein the recognition domain comprises a full-length antibody, a single-domain antibody, a recombinant heavy-chain-only antibody (VHH), a single-chain antibody (scFv), a shark heavy-chain-only antibody (VNAR), a microprotein (e.g. cysteine knot protein, knottin), a darpin, an anticalin, an adnectin, an aptamer, a Fv, a Fab, a Fab′, a F(ab′) 2 , a peptide mimetic molecule, a natural ligand for a receptor, or a synthetic molecule. 
     
     
         8 . The chimeric protein of any one of the above claims, wherein the recognition domain functionally modulates an antigen or receptor of interest. 
     
     
         9 . The chimeric protein of any one of the above claims, wherein the recognition domain recognizes and binds but does not functionally modulate an antigen or receptor of interest. 
     
     
         10 . The chimeric protein of any one of the above claims, wherein the modified signaling agent comprises one or more mutations conferring reduced affinity or activity at the signaling agent's receptor relative to a wild type signaling agent. 
     
     
         11 . The chimeric protein of any one of the above claims, wherein the modified signaling agent comprises one or more mutations conferring substantially reduced or ablated affinity or activity for a receptor relative to a wild type signaling agent. 
     
     
         12 . The chimeric protein of any one of the above claims, wherein the modified signaling agent comprises both (a) one or more mutations conferring substantially reduced or ablated affinity for a receptor relative to a wild type signaling agent and (b) one or more mutations conferring reduced affinity or activity for a receptor relative to a wild type signaling agent; and wherein the receptors are different. 
     
     
         13 . The chimeric protein of  claim 10 , wherein the one or more mutations allow for attenuation of activity. 
     
     
         14 . The chimeric protein of  claim 13 , wherein agonistic or antagonistic activity is attenuated. 
     
     
         15 . The chimeric protein of  claim 13  or  14 , wherein the modified signaling agent comprises one or more mutations which convert its activity from agonistic to antagonistic. 
     
     
         16 . The chimeric protein of  claim 10 , wherein the mutation confers reduced affinity or activity that is restorable by attachment to one or more targeting moiety. 
     
     
         17 . The chimeric protein of  claim 11 , wherein the mutation confers substantially reduced or ablated affinity or activity that is not substantially restorable by attachment to one or more targeting moiety. 
     
     
         18 . The chimeric protein of any one of the above claims, wherein the modified signaling agent is selected from one or more of an interferon, an interleukin, and a tumor necrosis factor. 
     
     
         19 . The chimeric protein of any one of the above claims, wherein the signaling agent is an interferon. 
     
     
         20 . The chimeric protein of any one of the above claims, wherein the chimeric protein is suitable for use in a patient having one or more of: cancer, infections, immune disorders, autoimmune diseases, cardiovascular diseases, wound, ischemia-related diseases, neurodegenerative diseases, and/or metabolic diseases. 
     
     
         21 . A chimeric protein comprising:
 (a) a first targeting moiety comprising a recognition domain which recognizes and binds to SIRP1α;   (b) a second targeting moiety comprising a recognition domain which recognizes and binds to PD-1; and   (b) a human interferon alpha 2 having one or more mutations that confer reduced affinity or activity at the signaling agent's receptor as compared to a wild type signaling agent, optionally selected from L153A, R149A, M148A, R144X 1 , A145X 2 , R33A, wherein   X 1  is selected from A, S, T, Y, L, and I,   X 2  is selected from G, H, Y, K, and D, and   wherein the targeting moiety and modified signaling agent are optionally connected with one or more linkers.   
     
     
         22 . A recombinant nucleic acid composition encoding one or chimeric proteins of any one of the above claims. 
     
     
         23 . A host cell comprising the nucleic acid of  claim 22 . 
     
     
         24 . A method for treating cancer, comprising administering an effective amount of the chimeric protein of any of the above claims to a patient in need thereof. 
     
     
         25 . The method of  claim 24 , wherein the cancer is selected form one or more of basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer (e.g., small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); melanoma; myeloma; neuroblastoma; oral cavity cancer (lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulval cancer; lymphoma including Hodgkin's and non-Hodgkin's lymphoma, as well as B-cell lymphoma (including low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; as well as other carcinomas and sarcomas; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (e.g. that associated with brain tumors), and Meigs' syndrome. 
     
     
         26 . The method of any one of  claim 24  or  25 , wherein the cancer overexpresses a Myc protein. 
     
     
         27 . The method of any one of  claims 24 - 26 , wherein the method induces and/or enhances phagocytosis of a cancer cell by macrophages. 
     
     
         28 . The chimeric protein of any one of the above claims for use as a medicament. 
     
     
         29 . The chimeric protein of any one of the above claims for use in the treatment of cancer. 
     
     
         30 . The chimeric protein of any one of the above claims for use in the treatment of hepatitis. 
     
     
         31 . Use of a chimeric protein of any one of the above claims in the manufacture of a medicament.