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(12) United States Patent (10) Patent No.: US 8,338,379 B2 Yamaguchi Et Al

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(12) United States Patent (10) Patent No.: US 8,338,379 B2 Yamaguchi Et Al US008338379B2 (12) United States Patent (10) Patent No.: US 8,338,379 B2 Yamaguchi et al. (45) Date of Patent: Dec. 25, 2012 (54) MONOCLONAL ANTIBODY AND USE Vandermeeren et al., “The Functional Y-Secretase Inhibitor Prevents THEREOF Production of Amyloid f 1-34 in Human Murine Cell Lines”. Neuroscience Letters, vol. 315, No. 3, pp. 145-148 (2001). Matthews et al., “Alzheimer's Disease-Related Overexpression of (75) Inventors: Haruyasu Yamaguchi, Maebashi (JP); the Cation-Dependent Mannose 6-Phosphate Receptor Increases AB Noriaki Kinoshita, Takasaki (JP); Secretion'. The Journal of Biological Chemistry, vol. 277, No. 7, pp. Masahiro Maeda, Takasaki (JP); Yuko 5299-5307 (2002). Horikoshi, Takasaki (JP) Janus, Christopher et al. "AB Peptide Immunization Reduces Behavioural Impairment and Plaques in a Model of Alzheimer's (73) Assignee: Intellect Neurosciences, Inc., New York, Disease”, Letters to Nature, vol. 408, No. 6815, pp. 978-983 (2000). NY (US) Tamaoka, Akira, "Alzhemer-Byo no Kotal Ryoho'. Gendal Iryo, vol. 34, No. 1, pp. 237-244 (2002). (*) Notice: Subject to any disclaimer, the term of this Kinebuchi et al., “A novel Cell Surface Antigen Involved in patent is extended or adjusted under 35 Thymocyte and Thymic Epithelial Cell Adhesion'. The Journal of U.S.C. 154(b) by 0 days. Immunology, vol. 146, No. 11, pp. 3721-3728 (1991). Frenkel et al., “High Affinity Binding of Monoclonal Antibodies to the Sequential Epitope EFRH of Beta-amyloid Peptide is Essential (21) Appl. No.: 12/888,661 for Fibrillar Aggregation.” Journal of Neuroimmunology, 95.136 142 (1999). (22) Filed: Sep. 23, 2010 Horikoshi et al., “Development of A-beta Terminal End-specific Antibodies and Sensitive ELISA for A-beta Variant.” Biochemical (65) Prior Publication Data and Biophysical Research Communications 319:733-737 (2004). US 2011 FOOO8339 A1 Jan. 13, 2011 Yamaguchi et al., “Diffuse Plaques Associated With Astroglial Amyloid Beta Protein, Possibly Showing a Disappearing Stage of Related U.S. Application Data Senile Plaques.” Acta Neuropathol 95:217-222 (1998). Morishima-Kawashima et al., “Effect of Apollipoprotein E Allele e4 (63) Continuation of application No. 10/589.969, filed as on the Initial Phase of Amyloid f-Protein Accumulation in the application No. PCT/JP2004/013536 on Sep. 16, Human Brain'. American Journal of Pathology, vol. 157, No. 6, pp. 2004, now Pat. No. 7,807,157. 2093-2099 (2000). Erlanger et al., “Steroid-Protein Conjugates'. Departments of (30) Foreign Application Priority Data Microbiology, Biochemistry, and Obstetrics and Gynecology, Col lage of Physicans, Columbia University, vol. 234, No. 5, pp. 1090 1094 (1958). Feb. 20, 2004 (JP) ................................. 2004-0451 11 Karu et al., “Synthesis of Haptens and Derivation of Monoclonal Antibodies for Immunoassay of the Phenylurea Herbicide Dluron'. J. (51) Int. Cl. Agric. Food Chem... vol. 42, No. 2, pp. 301-309 (1994). C07K 16/00 (2006.01) Yazaki et al., “Humanization of the Anti-CEA T84.66 Antibody AOIN37/18 (2006.01) Based on Crystal Structure Data'. Protein Engineering. Design & (52) U.S. Cl. ................... 514/17.8: 514/17.7:530/388.1; Selection, vol. 17, No. 5, pp. 481-489 (2004). 530/388.25; 530/387.3 Carter et al., “Humanization of an Anti-p185HER2 Antibody for (58) Field of Classification Search ........................ None Human Cancer Therapy”. Proc. Natl. Acad. Sci., vol. 89, pp. 4285 4289 (1992). See application file for complete search history. Tobinasiet al., “Feasibility and Pharmacokinetic Study of a Chimeric Anti-CD-20 Monoclonal Antibody (IDEC-C2B8, Rituximab) in (56) References Cited Relapsed B-Cell Lymphoma. The IDEC-C2B8 Study Group'. Ann. U.S. PATENT DOCUMENTS Oncol., vol. 9, No. 5, pp. 1-2 (1998). 5.437,995 A 8, 1995 Ichimori et al. Primary Examiner — Olga N Chernyshev 2003/OO73655 A1 4/2003 Chain 2005/O152903 A1 7/2005 Newman et al. (74) Attorney, Agent, or Firm — Fish & Richardson P.C. FOREIGN PATENT DOCUMENTS (57) ABSTRACT EP O125O23 11, 1984 An antibody capable of recognizing amyloid B while not EP O239.400 9, 1987 recognizing amyloid B precursor proteins, and a method for EP O683234 11, 1995 WO 9007861 7, 1990 using the same. WO 94.171.97 8, 1994 A monoclonal antibody characterized by being capable of WO 9420632 9, 1994 recognizing the N-terminus peptide of amyloid B while not WO OO72876 12/2000 recognizing amyloid B precursor proteins, an amyloid Bassay OTHER PUBLICATIONS kit, a therapeutic agent of Alzheimer's disease, and a method for treating Alzheimer's disease using the monoclonal anti Matthews et al., “Calpain Activity Regulates the Cell Surface Distri body. bution of Amyloid Precursor Protein'. The Journal of Biological Chemistry, vol. 277, No. 39, pp. 36415-36424 (2002). 10 Claims, 9 Drawing Sheets U.S. Patent Dec. 25, 2012 Sheet 1 of 9 US 8,338,379 B2 KDa 1. AB (1-40) 2. A B (1-42) 46.0 - 3. AB (1-43) 30.0 21.5 14.3 6.5 - FIG. I. U.S. Patent Dec. 25, 2012 Sheet 2 of 9 US 8,338,379 B2 ontrol" (42)AR-AB-AB-AB opes) Complex of 1C3 antibody and AB(1-42) FIG 2 U.S. Patent Dec. 25, 2012 Sheet 3 of 9 US 8,338,379 B2 A 8(1-40) A B(2-40) AB(3-40) O 10 1 00 250 1 0 1 00 250 pg FIG 3 U.S. Patent Dec. 25, 2012 Sheet 4 of 9 US 8,338,379 B2 A B Sapple dilution x1 x2 x 10 x2 x 0 -- APP FIG 4 U.S. Patent Dec. 25, 2012 Sheet 5 Of 9 US 8,338,379 B2 O 2O 40 60 BO OO 20 AB(1-40) concentration (pg/ml) O 2O 40 60 80 100 120 AB(1-42) concentration (pg/ml) FIG 5 U.S. Patent Dec. 25, 2012 Sheet 6 of 9 US 8,338,379 B2 FIG 6 U.S. Patent Dec. 25, 2012 Sheet 7 Of 9 US 8,338,379 B2 FIG 7 U.S. Patent Dec. 25, 2012 Sheet 8 of 9 US 8,338,379 B2 Mouse to which 82E1 antibody was administered 8 8 Control mouse AB content (%) Cortical layer Hippocampus FIG 8 U.S. Patent Dec. 25, 2012 Sheet 9 Of 9 US 8,338,379 B2 o Mouse to which 82E antibody was administered e Control mouse * PCO.05 A 3 (1-42) A 3 (x-42) FIG 9 US 8,338,379 B2 1. 2 MONOCLONAL ANTIBODY AND USE In addition, a method of Suppressing accumulation of amy THEREOF loid B in the brain in order to prevent Alzheimer's disease has been investigated. However, since the presently known amy CROSS REFERENCE TO PRIORAPPLICATIONS loid 3 antibodies react not only with an amyloid B, but also APP in normal cells, these antibodies have high possibility of This application is a continuation application of U.S. Ser. causing side reaction Such as inflammation and, therefore, No. 10/589,969, filed Aug. 18, 2006, which is the U.S. have not been clinically applied. National Phase under 35 U.S.C. S371 of International Patent Patent Document 1: WO 1994/017.197 pamphlet Application No. PCT/JP2004/013536, filed Sep. 16, 2004, and claims the priority of Japanese Patent Application No. 10 DISCLOSURE OF THE INVENTION 2004-0451 11, filed Feb. 20, 2004. Each of the aforemen tioned applications is hereby incorporated herein by refer Problems to be Solved by the Invention ence in its entirety. The International Application published in Japanese on Sep. 1, 2005 as WO 2005/080435 under PCT 15 Accordingly, a first object of the present invention is to Article 21(2). provide an antibody capable of recognizing an amyloid B, but not APP. REFERENCE TO SEQUENCE LISTING A second object of the present invention is to provide a system for precisely assaying amyloid 3 (1-40) and amyloid Pursuant to 37 C.F.R. 1.821(c), a sequence listing is sub B (1-42) which completely possess the entire length, by using mitted herewith as an ASCII compliant text file named the antibody. “Sequence Listing..txt that was created on Sep. 23, 2010, and A third object of the present invention is to provide a has a size of 1,798 bytes. The content of the aforementioned therapeutic agent and the like for Alzheimer's disease com file named “Sequence Listing..txt is hereby incorporated by prising the above antibody as an active ingredient. reference in its entirety. 25 TECHNICAL FIELD Means for Solving the Problems The present invention relates to a monoclonal antibody As a result of extensive studies in order to solve the above which can recognize the N-terminus peptide of an amyloid B, first object, the inventors of the present invention have found 30 that an antibody which can recognize an amyloid B, but not but not an amyloid B precursor protein, and a method of using APP can be obtained by immunizing with a specific peptide. the monoclonal antibody. In addition, as a result of extensive studies in order to solve the above second object, the inventors have found that a BACKGROUND ART system that can precisely assay amyloid (1-40) and amyloid 3 An amyloid B is a peptide consisting of 40 or 42 amino 35 (1-42) which completely possess the entire length (the amy acids and is generated from an amyloid B precursor protein loid (1-40) and amyloid B (1-42) are hereinafter referred to (APP) cleaved by B-selectase and Y-selectase. The amyloid B from time to time collectively as "amyloids f”) can be con with 40 amino acids is referred to as amyloid f (1-40) and the structed by using the above antibody.
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