US 2007008 7001A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0087001 A1 Taylor et al. (43) Pub. Date: Apr. 19, 2007 (54) INHIBITION OF PLACENTAGROWTH Publication Classification FACTOR (PLGF). MEDIATED METASTASIS AND/OR ANGIOGENESIS (51) Int. Cl. A 6LX 39/395 (2006.01) (75) Inventors: Alice P. Taylor, Rockaway, NJ (US); A6II 38/18 (2006.01) David M. Goldenberg, Belleville, NJ (52) U.S. Cl. .............................. 424/145.1: 514/12: 514/9 (US); Chien Hsing Chang, Downington, PA (US) (57) ABSTRACT Correspondence Address: The present invention concerns methods and compositions FAEGRE & BENSON LLP for inhibiting angiogenesis and/or tumor growth, Survival PATENT DOCKETING and/or metastasis. In particular embodiments, the methods 22OO WELLS FARGO CENTER and compositions may concern ligands against placenta 90 SOUTH SEVENTH STREET growth factor (PIGF), such as BP-1, BP-2, BP-3 or BP-4. MINNEAPOLIS, MN 55402-3901 (US) Some methods may comprise administering one or more PlGF ligands, alone or in combination with one or more (73) Assignees: CENTER FOR MOLECULARMEDI other agents, such as chemotherapeutic agents, other anti CINE AND IMMUNOLOGY: IMMU angiogenic agents, immunotherapeutic agents or radioim NOMEDICS, INC. munotherapeutic agents to a Subject. The PlGF ligands are (21) Appl. No.: 111581,287 effective to inhibit angiogenesis, tumor cell motility, tumor metastasis, tumor growth and/or tumor Survival. In certain (22) Filed: Oct. 16, 2006 embodiments, PlGF ligands may be administered to subjects to ameliorate other angiogenesis related conditions. Such as Related U.S. Application Data macular degeneration. In some embodiments, PIGF expres sion levels may be determined by any known method to (60) Provisional application No. 60/728.292, filed on Oct. select those patients most likely to respond to PIGF targeted 19, 2005. therapies. 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O O Days Patent Application Publication Apr. 19, 2007 Sheet 6 of 6 US 2007/0087.001 A1 9eun61-I ZTHpdo-I-OZCIO?S US 2007/0087.001 A1 Apr. 19, 2007 INHIBITION OF PLACENTA GROWTH FACTOR ing, Suppressing, blocking and/or eliminating angiogenesis (PLGF) MEDIATED METASTASIS AND/OR and/or tumor metastasis. In certain embodiments, the com ANGOGENESIS positions and/or methods may concern ligands for placenta growth factor-2 (P1GF), and/or ligands binding to both Flt-1 RELATED APPLICATIONS and PlGF. Such ligands may include, but are not limited to, 0001. The present application claims the benefit under 35 peptides, antibodies, antibody fragments, humanized anti U.S.C. S 119(e) of provisional U.S. Patent Application Ser. bodies, chimeric antibodies, antibody analogs, aptamers, No. 60/728,292, filed on Oct. 19, 2005, the entire text of organic compounds and/or any other molecules or com which is incorporated herein by reference. pounds known in the art that are ligands for PlGF. 0009. In various embodiments, the PlGF ligands may be FEDERALLY FUNDED RESEARCH peptides. Methods for identifying peptides that bind to 0002 The studies disclosed herein were supported in part particular targets are well known in the art, including for by grant DAMD 17-03-1-0758 from the U.S. Department of example phage display techniques as discussed below. In Defense (A.P.T.). The U.S. government may have certain phage display, libraries of peptides expressed on the Surface rights to practice the Subject invention. of phage. Such as filamentous bacteriophage, are created and may be screened by selection against the target(s) of interest. BACKGROUND After one or more rounds of Screening (panning) against the target, phage containing display peptides that bind to the 0003) 1. Field of the Invention target may be isolated and the peptide sequences deter 0004 The present invention concerns methods and com mined, for example by sequencing of the peptide-encoding positions for inhibiting angiogenesis, particularly pathologic DNA inserts in the phage nucleic acid. angiogenesis and/or growth and metastasis of tumors. In particular embodiments, the compositions and methods con 0010. Other embodiments concern methods and/or com cern inhibitors targeted to placenta growth factor-2 (PlGF) positions for treating Subjects, such as Subjects with cancer and/or its receptor, Flt-1, including but not limited to pep and/or with a condition related to angiogenesis (e.g., macu tides, antibodies, antibody fragments, humanized antibodies, lar degeneration, etc.). Subjects may include, but are not chimeric antibodies, antibody analogs, aptamers, organic limited to, humans, animals, cats, dogs, cows, sheep, goats, compounds and/or any other molecule or compound known horses, alpacas and mammals. The methods and composi in the art that may be used to inhibit PlGF-mediated angio tions may comprise one or more PlGF ligands to be admin genesis and/or metastasis. In more particular embodiments, istered to a subject. In preferred embodiments, peptides the inhibitors may be peptides isolated by phage display identified as ligands by phage display against PlGF may be against PlGF. administered. Administration may be by any route known in the art, Such as oral, nasal, buccal, inhalational, rectal, 0005 2. Description of Related Art vaginal or topical. Alternatively, administration may be by 0006 The high expression of angiogenic growth factors orthotopic, intradermal, Subcutaneous, intramuscular, intra is necessary for the growth and development of diverse peritoneal, intraarterial, intrathecal or intravenous injection. cancers (Cao et al., 1998, J Clin Invest 101:1055-1063). In preferred embodiments, the PlGF ligand (inhibitor) is Among Such angiogenic growth factors, high expression of orally administered. In more preferred embodiments, the vascular endothelial growth factor (VEGF) has received the PlGF ligand may comprise the sequence of (binding protein) most attention with regard to being associated with BP-1, BP-2, BP-3 or BP-4, as disclosed in the Examples increased malignancy and metastasis (Abdulrauf et al., below. 1998, J Neurosurg88:513-52: Ahmed et al., 2000, Placenta 0011. The administration of PlCF ligands may be effec 21 Suppl A, S16-24). However, the role of VEGF and other tive to inhibit or eliminate tumor metastasis, angiogenesis in members of the VEGF family of growth factors in tumor solid tumors, tumor cell survival and/or tumor cell mobility. recurrence or metastatic growth has not been clearly estab In preferred embodiments, administration of one or more lished. Many treatments, including radiation, cytokines, and PlGF ligands may prevent tumors from metastasizing or cytotoxic agents, owe Some of their therapeutic activity to may result in regression or growth inhibition of existing anti-angiogenic mechanisms, one of which is the down tumors. The skilled artisan will realize that one or more regulation of VEGF (Taylor et al., 2002a, Clin Cancer Res PlGF ligands may be administered alone or alternatively in 8:1213-1222; Jiang et al., 1999, Mol Carcinog 26:213-225; conjunction with other known therapeutic treatments for Machein et al., 1999, Neuropathol Appl Neurobiol 25:104 cancer and/orangiogenesis, such as chemotherapy, radiation 112). Yet, marked inhibition of VEGF does not always result therapy, immunotherapy, anti-VEGF agents and/or other in durable responses or even cure. This suggests that factors known anti-angiogenesis agents and the like. In some other than VEGF may be capable of restoring a blood supply embodiments, the PlGF ligands may be administered with a to Surviving tumor cells. bispecific antibody, with one binding site for the PlGF ligand 0007. A need exists in the field for methods and compo and a second binding site for a tumor antigen or other target. sitions directed towards the effective inhibition of tumor In other embodiments, PlGF ligands may be covalently angiogenesis and/or growth and metastasis, as well as angio attached to or provided as a fusion protein with an antibody, antibody fragment, monoclonal antibody, Fc fragment, Fc genesis associated with other diseases. binding protein or antibody binding protein. SUMMARY OF THE INVENTION 0012. In alternative embodiments, PIGF ligands may be 0008. The present invention fulfills an unresolved need in of use to treat angiogenesis related conditions besides can the art by providing methods and compositions for inhibit cer. Examples of conditions related to angiogenesis include, US 2007/0087.001 A1 Apr. 19, 2007 but are not limited to, rheumatoid arthritis, inflammatory results-SD (n=3-5 experiments). Abbreviations used in the bowel disease, Crohn's disease, ulcerative colitis, sarcoido graph: P1 is PIGF; “VE is VEGF. *P-0.05 (ANOVA). sis, asthma, edema, pulmonary hypertension, formation and development of tumors, psoriasis, diabetic retinopathy, 0018 FIG. 3. BP1 inhibits MDA-MB-231 xenograft macular degeneration, corneal graft rejection, neovascular growth and metastasis. FIG. 3A. Tumor Volume vs. time, s.c. glaucoma, myocardial angiogenesis, plaque neovasculariza model. Results shown are from one of two experiments+SD tion, restenosis, neointima formation after vascular trauma, (n=3-4 mice/treatment).