USOO9234015B2 (12) United States Patent (10) Patent No.: US 9.234,015 B2 Sentissi et al. (45) Date of Patent: Jan. 12, 2016 (54) CHLOROTOXIN POLYPEPTIDES AND 6,028,174 A 2/2000 Ullrich et al. CONUGATES AND USES THEREOF 6,130, 101 A 10/2000 Mao et al. 6,319,891 B1 1 1/2001 Sontheimer et al. 6,429,187 B1 8, 2002 Sontheimer et al. (71) Applicant: MORPHOTEK, INC., Exton, PA (US) 6,514,481 B1 2/2003 Prasadet al. 6,555,652 B1 4/2003 Itoh et al. (72) Inventors: Abdellah Sentissi, Dover, MA (US); 6,667,156 B2 12/2003 Lyons et al. Douglas B. Jacoby, Wellesley, MA (US) 6,767,635 B1 7/2004 Bahr et al. 6,849,714 B1 2/2005 Bridon et al. (73) Assignee: Morphotek, Inc., Exton, PA (US) 6,926,8966,870,029 B2 3/20058, 2005 SontheimerBosslet et al. et al. 6,972,326 B2 12/2005 Haugland et al. (*) Notice: Subject to any disclaimer, the term of this 7,094,868 B2 8, 2006 SNE f al. patent is extended or adjusted under 35 7,252,998 B2 8, 2007 Skerra et al. U.S.C. 154(b) by 0 days. 7.462.446 B2 12/2008 Zhang et al. 7,678,759 B2 3/2010 Sontheimer et al. 8,227,439 B2 7, 2012 O'Neill et al. (21) Appl. No.: 14/450,182 8,470,607 B2 6/2013 Jacoby et al. 8,778,310 B2 7/2014 Zhang et al. (22) Filed: Aug. 1, 2014 2001/0007025 A1 7/2001 Bennett et al. 2001/0055751 A1 12/2001 Reiter et al. (65) Prior Publication Data 2002/0065216 A1 5/2002 Sontheimer et al. 2002/0146749 A1 10/2002 Lyons PH.D. et al. US 2015/OO30537 A1 Jan. 29, 2015 2003/0021810 A1 1/2003 Sontheimer et al. 2003/0201208 A1 10, 2003 Koch et al. 2003/0216322 A1 1 1/2003 Samoylova et al. Related U.S. Application Data 2004/0101822 A1 5, 2004 Wiesner et al. 2004/0102381 A1 5/2004 Ekwuribe et al. (63) Continuation of application No. 13/577,169, filed as 2004/0105.980 A1 6/2004 Sudarshan et al. application No. PCT/US2011/023823 on Feb. 4, 2011, (Continued) now Pat. No. 9,018,347. (60) Provisional application No. 61/301,615, filed on Feb. FOREIGN PATENT DOCUMENTS 4, 2010. CN 1924006. A 3, 2007 CN 1010O3788 A 7/2007 (51) Int. Cl. CN 1O127O158. A 9, 2008 A6 IK38/00 (2006.01) CN 101381405 A 3, 2009 C07K I4/435 (2006.01) CN 101824084 A 9, 2010 A6 IK 47/48 (2006.01) CN 101921769. A 12/2010 C07K 14/00 (2006.01) N ISSA, 35. A6 IK3I/337 (2006.01) EP 1430.131 A2 6, 2004 A6 IK5I/08 (2006.01) EP 2182004 A1 5.2010 (52) U.S. Cl. (Continued) CPC ......... C07K 14/.43522 (2013.01); A61 K3I/337 (2013.01); A61 K47/48215 (2013.01); A61 K OTHER PUBLICATIONS 47/48261 (2013.01); A61 K5I/088 (2013.01); A61 K38/00 (2013.01) Akabani et al., Dosimetry and Radiographic Analysis of 131 I-La (58) Field of Classification Search beled Anti-Tenascin 81C6 Murine Monoclonal Antibody in Newly CPC ............ A61 K38/00; A61K 47/48215: A61 K Diagnosed Patients with Malignant Gliomas: A Phase II Study, Jour 51/088: A61 K31/337; A61K 47/48261; nal of Nuclear Medicine, 46(6):1042-1051 (2005). CO7K 14/43522 (Continued) See application file for complete search history. (56) References Cited Primary Examiner — Julie Ha U.S. PATENT DOCUMENTS Assistant Examiner — Li Ni Komatsu (74) Attorney, Agent, or Firm — Brenda Herschbach Jarrell; 4,444,744 A 4/1984 Goldenberg Choate Hall & Stewart LLP 5,051,364 A 9, 1991 Isacke et al. 5,212.290 A 5/1993 Vogelstein et al. 5,223,253 A 6, 1993 Hall et al. 5,236,844 A 8, 1993 Basset et al. (57) ABSTRACT 5,314,992 A 5/1994 Guyre et al. 5,591,829 A 1/1997 Matsushita Reduced lysine chlorotoxin polypeptides that may be used to 5,626,862 A 5, 1997 Brem et al. generate single species conjugates of chlorotoxin. Conju 5,688,773 A 11/1997 Chiocca et al. gates comprising Such chlorotoxin polypeptides and pharma 5,750,376 A 5, 1998 Weiss et al. 5,756,340 A 5, 1998 Hammocket al. ceutical compositions thereof. Methods of using such com 5,866,570 A 2/1999 Liang et al. positions and/or conjugates. 5,905,027 A 5, 1999 Ullrich et al. 5,935,795 A 8, 1999 Lin et al. 5,985,822 A 11/1999 Edelman et al. 16 Claims, No Drawings US 9.234,015 B2 Page 2 (56) References Cited WO WO-2009052390 A1 4/2009 WO WO-2009052392 A1 4/2009 U.S. PATENT DOCUMENTS WO WO-2009052400 A1 4/2009 WO WO-2009062520 A1 5, 2009 2004/O141981 A1 T/2004 Sontheimer et al. WO WO-2009.140599 6, 2009 2004/0180846 A1 9, 2004 Huang et al. WO WO-2009 108762 A2 9, 2009 2005. O142062 A1 6, 2005 Sontheimer et al. WO WO-2009 117018 A1 9, 2009 2005/0261191 A1 11, 2005 Barasch et al. WO WO-2009 15645.6 A1 12/2009 2006, OO1925.6 A1 1, 2006 Clarke et al. WO WO-2O11057295 A2 5, 2011 2006, OO19347 A1 1, 2006 Cho et al. WO WO-2011/097533 A1 8, 2011 2006, OO88899 A1 4, 2006 Alvarez et al. WO WO-2011094671 A2 8, 2011 2006, O166892 A1 T/2006 Alvarez et al. WO WO-2011 142858 A2 11/2011 2007/007 1764 A1 3, 2007 Sullivan et al. WO WO-2012/O22742 A1 2, 2012 2007/O154965 A1 7/2007 Zhang et al. 2007/0237714 A1 10, 2007 Alvarez OTHER PUBLICATIONS 2007/0275902 A1 11/2007 Gonda et al. 2008. O153746 A1 6, 2008 Alvarez et al. Akabani et al., Dosimetry of 131 I-Labeled 81C6 Monoclonal Anti 2008/0279780 A1 11/2008 Zhang et al. body Administered into Sugically Created Resection Cavities in 2009 OOO4105 A1 1/2009 Cheng et al. Patients with Malignant Brain Tumors, J of Nucl. Med., 40:631-638 2009, O12394.6 A1 5/2009 Birkenmeyer et al. 2009/O123970 A1 5/2009 Tu et al. (1999). 2009, O124022 A1 5/2009 Birkenmeyer et al. Amersham Biosciences Publication, Labelling of Proteins with 2009/0176274 A1 T/2009 Tu et al. CyOye N-hydroxysuccinimide Esters for Fluorescent Applications 2009/02O3598 A1 8, 2009 McCarty et al. on the LEAOSeeker Homogeneous Imaging System, Leadseeker 2009,0263894 A1 10, 2009 Birkenmeyer et al. L8:1-4, (2001). 2009,0269777 A1 10, 2009 Birkenmeyer et al. Applebaumet al., Treatment of Malignant Lymphoma in 100 Patients 2009/03O4592 A1 12, 2009 O'Neill et al. With Chemotherapy, Total Body Irradiation, and Marrow Transplan 20090311224 A1 12, 2009 Lee et al. tation, J. Clin. Oncol. 5:1340-1341 (1987). 2010/0098637 A1 4, 2010 Orringer et al. Baker, Effects of an epithelial CI channel blocker on whole cell 2010/0105150 A1 4, 2010 Adamczyk et al. voltage clamp and patch clamp recordings from a human astrocytoma 2010/0210546 A1 8, 2010 Alvarez et al. in culture, J. Physiol. 438: 128-129 (1991). 2010/0215575 A1 8, 2010 O'Neill et al. Bertolini et al., Inhibition of angiogenesis and induction of 2010/0215576 A1 8, 2010 Sontheimer et al. endothelial and tumor cell apoptosis by green tea in animal models of 2011/0027177 A1 2, 2011 Jacoby et al. human high-grade non-Hodgkin's lymphoma, Leukemia, 14:1477 2011/009 1380 A1 4, 2011 Jacoby et al. 1482 (2000). 2011/0311445 A1 12/2011 Alvarez et al. Binger et al., Journal of Clinical Oncology, 16(6): 2202-2212 (1998). 2012/01561.31 A1 6, 2012 Alvarez Bodey et al., Failure of CancerVaccines: The Significant Limitations 2012/0183544 A1 T/2012 Sontheimer et al. of this Approach to Immunotherapy, Anticancer Res., 20: 2665-2667 2013,0028836 A1 1, 2013 Sentissi et al. (2000). 2013,0045163 A1 2, 2013 O'Neill et al. Bowie et al., Deciphering the Message in Protein Sequences: Toler 2013/O19576.0 A1 8, 2013 Olson ance to Amino Acid Substitutions, Science, 257: 1306-1310 (1990). 2014/024.1993 A1 8, 2014 Zhang et al. Brem et al., Placebo-controlled trial of safety and efficacy of 2015, OO10473 A1 1/2015 Alvarez intraoperative controlled delivery by biodegradable polymers of che motherapy for recurrent gliomas, The Lancet, 345:1008-1012 FOREIGN PATENT DOCUMENTS (1995). JP HO8-325291 A 12/1996 Brismar, T. and Collins, V., Inward rectifying potassium channels in JP HO9-71559 A2 3, 1997 human malignant glioma cells, Brain Res. 480:249-258 (1989). JP 2008-538506 A 10, 2008 Brismar, T. and Collins, V. Potassium and sodium channels in human WO WO-8802.117 A1 3, 1988 malignant glioma cells, Brain Res. 480:259-267 (1989). WO WO-9311222 A1 6, 1993 Britton et al., Prostate cancer: the contribution of nuclear medicine, WO WO-9415615 A1 T 1994 BJU Int’l 86(1):135-142 (2000). WO WO-9724619 A1 7/1997 Burger et al., Topographic anatomy and CT correlations in the WO WO-99.297 15 A1 6, 1999 untreated glioblastoma multiforme, J. Neurosurg, 68:698-704 WO WO-00,6281.0 A1 10, 2000 (1988). WO WO-0062807 A1 10, 2000 Burgess et al., Possible Dissociation of the Heparin-binding and WO WO-03/008583 A2 1, 2003 Mitogenic Activities of Herparin-binding (Acidic Fibroblast) WO WO-03000203 A2 1, 2003 WO WO-03101474 A1 12/2003 Growth Factor-1 from Its Receptor-binding Activities by Site-di WO WO-O3101475 A1 12/2003 rected Mutagenesis of a Single Lysine Residue, J Cell Biol.