Fragments of p97 and uses thereof

Abstract

Provided are fragments of human p97 (melanotransferrin) polypeptides having blood-brain barrier (BBB) transport activity, including variants and combinations thereof, conjugates comprising the p97 fragments, and related methods of use thereof, for instance, to facilitate the delivery of therapeutic or diagnostic agents across the BBB and into the central nervous system.

1 . An isolated p97 polypeptide of up to about 300 amino acids in length, where the polypeptide comprises an amino acid sequence at least 70% identical to DSSHAFTLDELR (SEQ ID NO:13) or any one or more of SEQ ID NO:2-18. 
     
     
         2 . The isolated p97 polypeptide of  claim 1 , where the polypeptide comprises DSSHAFTLDELR (SEQ ID NO:13) or one or more of SEQ ID NO:2-18, optionally including adjacent C-terminal and/or N-terminal sequences as defined by SEQ ID NO:1. 
     
     
         3 . The isolated p97 polypeptide of  claim 1 , where the polypeptide comprises 2, 3, 4, or 5 of DSSHAFTLDELR (SEQ ID NO:13) or SEQ ID NOS:2-18, optionally including any intervening sequences as defined by SEQ ID NO:1. 
     
     
         4 . The isolated p97 polypeptide of  claim 1 , where the polypeptide comprises one or both of SEQ ID NO:13 and/or 14, optionally including intervening sequences as defined by SEQ ID NO:1. 
     
     
         5 . The isolated p97 polypeptide of  claim 1 , where the polypeptide is up to about 250, 200, 150, 100, 50, 20, or 10 amino acids in length. 
     
     
         6 . The isolated p97 polypeptide of  claim 1 , where the polypeptide is fused to a heterologous protein. 
     
     
         7 . A conjugate, comprising the p97 polypeptide of  claim 1 , where the p97 polypeptide is covalently or operatively linked to an agent, to form a p97-agent conjugate. 
     
     
         8 . The conjugate of  claim 7 , where the agent is a small molecule, a polypeptide, a peptide mimetic, a peptoid, an aptamer, or a detectable entity. 
     
     
         9 . The conjugate of  claim 8 , where the small molecule is a cytotoxic or chemotherapeutic or anti-angiogenic agent selected from one or more of alkylating agents, anti-metabolites, anthracyclines, anti-tumor antibiotics, platinums, type I topoisomerase inhibitors, type II topoisomerase inhibitors, vinca alkaloids, and taxanes. 
     
     
         10 . The conjugate of  claim 8 , where the small molecule is selected from one or more of chlorambucil, cyclophosphamide, cilengitide, lomustine (CCNU), melphalan, procarbazine, thiotepa, carmustine (BCNU), enzastaurin, busulfan, daunorubicin, doxorubicin, gefitinib, erlotinib idarubicin, temozolomide, epirubicin, mitoxantrone, bleomycin, cisplatin, carboplatin, oxaliplatin, camptothecins, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, teniposide, temsirolimus, everolimus, vincristine, vinblastine, vinorelbine, vindesine, CT52923, paclitaxel, imatinib, dasatinib, sorafenib, pazopanib, sunitnib, vatalanib, geftinib, erlotinib, AEE-788, dichoroacetate, tamoxifen, fasudil, SB-681323, semaxanib, donepizil, galantamine, memantine, rivastigmine, tacrine, rasigiline, naltrexone, lubiprostone, safinamide, istradefylline, pimavanserin, pitolisant, isradipine, pridopidine (ACR16), tetrabenazine, bexarotene, glatirimer acetate, fingolimod, and mitoxantrone, including pharmaceutically acceptable salts and acids thereof. 
     
     
         11 . The conjugate of  claim 8 , where the polypeptide is an antibody or antigen-binding fragment thereof. 
     
     
         12 . The conjugate of  claim 11 , where the antibody or antigen-binding fragment thereof specifically binds to a cancer-associated antigen. 
     
     
         13 . The method of  claim 12 , where the cancer-associated antigen is one or more of human Her2/neu, Her1/EGF receptor (EGFR), Her3, A33 antigen, B7H3, CD5, CD19, CD20, CD22, CD23 (IgE Receptor), C242 antigen, 5T4, IL-6, IL-13, vascular endothelial growth factor VEGF (e.g., VEGF-A) VEGFR-1, VEGFR-2, CD30, CD33, CD37, CD40, CD44, CD51, CD52, CD56, CD74, CD80, CD152, CD200, CD221, CCR4, HLA-DR, CTLA-4, NPC-1C, tenascin, vimentin, insulin-like growth factor 1 receptor (IGF-1R), alpha-fetoprotein, insulin-like growth factor 1 (IGF-1), carbonic anhydrase 9 (CA-IX), carcinoembryonic antigen (CEA), integrin α v β 3 , integrin α 5 β 1 , folate receptor 1, transmembrane glycoprotein NMB, fibroblast activation protein alpha (FAP), glycoprotein 75, TAG-72, MUC1, MUC16 (or CA-125), phosphatidylserine, prostate-specific membrane antigen (PMSA), NR-LU-13 antigen, TRAIL-R1, tumor necrosis factor receptor superfamily member 10b (TNFRSF10B or TRAIL-R2), SLAM family member 7 (SLAMF7), EGP40 pancarcinoma antigen, B-cell activating factor (BAFF), platelet-derived growth factor receptor, glycoprotein EpCAM (17-1A), Programmed Death-1, protein disulfide isomerase (PDI), Phosphatase of Regenerating Liver 3 (PRL-3), prostatic acid phosphatase, Lewis-Y antigen, GD2 (a disialoganglioside expressed on tumors of neuroectodermal origin), glypican-3 (GPC3), or mesothelin. 
     
     
         14 . The conjugate of  claim 11 , where the antibody or antigen-binding fragment thereof specifically binds to a pain-associated antigen. 
     
     
         15 . The method of  claim 14 , where the pain associated-antigen is one or more of nerve growth factor (NGF) or tropomyosin-related kinase A (TrkA). 
     
     
         16 . The conjugate of  claim 11 , where the antibody or antigen-binding fragment thereof specifically binds to a pro-inflammatory molecule, optionally a pro-inflammatory cytokine or chemokine. 
     
     
         17 . The conjugate of  claim 16 , where the pro-inflammatory molecule is one or more of TNF-α, TNF-β, FasL, CD27L, CD30L, CD40L, Ox40L, 4-1BBL, TRAIL, TWEAK, and Apo3L, IL-1α, IL-1β, IL-2, interferon-γ (IFN-γ), IFN-α, IFN-β, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-21, LIF, CCL5, GROα, MCP-1, MIP-1α, MIP-1β, macrophage colony stimulating factor (MCSF), or granulocyte macrophage colony stimulating factor (GM-CSF). 
     
     
         18 . The conjugate of  claim 17 , where the pro-inflammatory molecule is TNF-α, and where the antibody is adalimumab, certolizumab pegol, etanercept, golimumab, infliximab, D2E7, CDP 571, or CDP 870, or an antigen-binding fragment or variant thereof. 
     
     
         19 . The conjugate of  claim 11 , wherein the antibody or antigen-binding fragment thereof specifically binds to one or more of human Her2/neu, Her1/EGFR, TNF-α, B7H3 antigen, CD20, VEGF, CD52, CD33, CTLA-4, tenascin, alpha-4 (α4) integrin, IL-23, amyloid-β, Huntingtin, CD25, nerve growth factor (NGF), TrkA, or α-synuclein. 
     
     
         20 . The conjugate of  claim 11 , where the antibody is selected from one or more of trastuzumab, cetuximab, daclizumab, tanezumab, 3F8, 8H9, abagovomab, adecatumumab, afutuzumab, alemtuzumab, alacizumab (pegol), amatuximab, apolizumab, bavituximab, bectumomab, belimumab, bevacizumab, bivatuzumab (mertansine), brentuximab vedotin, cantuzumab (mertansine), cantuzumab (ravtansine), capromab (pendetide), catumaxomab, citatuzumab (bogatox), cixutumumab, clivatuzumab (tetraxetan), conatumumab, dacetuzumab, dalotuzumab, detumomab, drozitumab, ecromeximab, edrecolomab, elotuzumab, enavatuzumab, ensituximab, epratuzumab, ertumaxomab, etaracizumab, farletuzumab, FBTA05, figitumumab, flanvotumab, galiximab, gemtuzumab, ganitumab, gemtuzumab (ozogamicin), girentuximab, glembatumumab (vedotin), ibritumomab tiuxetan, icrucumab, igovomab, indatuximab ravtansine, intetumumab, inotuzumab ozogamicin, ipilimumab (MDX-101), iratumumab, labetuzumab, lexatumumab, lintuzumab, lorvotuzumab (mertansine), lucatumumab, lumiliximab, mapatumumab, matuzumab, milatuzumab, mitumomab, mogamulizumab, moxetumomab (pasudotox), nacolomab (tafenatox), naptumomab (estafenatox), narnatumab, necitumumab, nimotuzumab, nivolumab, Neuradiab® (with or without radioactive iodine), NR-LU-10, ofatumumab, olaratumab, onartuzumab, oportuzumab (monatox), oregovomab, panitumumab, patritumab, pemtumomab, pertuzumab, pritumumab, racotumomab, radretumab, ramucirumab, rilotumumab, rituximab, robatumumab, samalizumab, sibrotuzumab, siltuximab, tabalumab, taplitumomab (paptox), tenatumomab, teprotumumab, TGN1412, ticilimumab, tremelimumab, tigatuzumab, TNX-650, tositumomab, TRBSO7, tucotuzumab (celmoleukin), ublituximab, urelumab, veltuzumab, volociximab, votumumab, and zalutumumab, including antigen-binding fragments thereof.