USOO86428O1B2

(12) United States Patent (10) Patent No.: US 8,642,801 B2 Kong et al. (45) Date of Patent: Feb. 4, 2014

(54) METHODS AND COMPOSITIONS FOR 4,528, 184 A 7, 1985 Kurono et al. TREATING AMYLOID-RELATED DISEASES 4,540,564 A 9, 1985 Bodor 4,563,470 A 1/1986 Durlach 4,593,045 A 6, 1986 Flork et al. (75) Inventors: Xianqi Kong, Dollard-des-Ormeaux 4,713,376 A 12/1987 Kuzuya et al. (CA); David Migneault, Laval (CA); 4,737,353 A 4, 1988 Flanigen et al. Isabelle Valade, Laval (CA); Xinfu Wu, 4,737,357 A 4, 1988 Lehmann et al. 4,812,512 A 3, 1989 Buendia et al. Laval (CA); Francine Gervais, Ile 4,847,082 A 7, 1989 Sabin Bizard (CA) 4.956,347 A 9, 1990 Ban et al. 5,017,566 A 5, 1991 Bodor (73) Assignee: BHI Limited Partnership, Laval, 5,023,252 A 6, 1991 HSeih Quebec (CA) 5,024,998 A 6, 1991 Bodor 5,039,794. A 8, 1991 Wier et al. Notice: Subject to any disclaimer, the term of this 5,064,923 A 11/1991 Kashihara et al. (*) 5, 112,863. A 5/1992 Hashimoto et al. patent is extended or adjusted under 35 5,124,146 A 6/1992 Neuwelt U.S.C. 154(b) by 190 days. 5,153,179 A 10, 1992 Eb1 5,164,295 A 1 1/1992 Kisilevsky et al. (21) Appl. No.: 12/613,412 5,166.320 A 11/1992 Wu et al. 5,177,064 A 1/1993 Bodor Filed: Nov. 5, 2009 5, 192,753 A 3, 1993 McGeer et al. (22) 5,194,654 A 3, 1993 Hostetler et al. 5,223,510 A 6/1993 Gubinet al. (65) Prior Publication Data 5,242.932 A 9/1993 Gandy et al. 5,254,342 A 10, 1993 Shen et al. US 2010/O113591 A1 May 6, 2010 5,258.402 A 1 1/1993 Maryanoff 5,270,312 A 12/1993 Glase et al. Related U.S. Application Data 5,276,059 A 1/1994 Caughey et al. 5,284,876 A 2, 1994 Shashoua et al. (63) Continuation of application No. 1 1/316,694, filed on 5,318,958 A 6/1994 Kisilevsky Dec. 21, 2005, now abandoned, which is a 5,342,977 A 8/1994 Baschang et al. continuation-in-part of application No. 10/871,514, 5,374,548 A 12/1994 Caras filed on Jun. 18, 2004, now Pat. No. 7,414,076. (Continued) (60) Provisional application No. 60/480,906, filed on Jun. FOREIGN PATENT DOCUMENTS 23, 2003, provisional application No. 60/512,047, filed on Oct. 17, 2003, provisional application No. CA 10953.06 A1 2, 1981 60/638,819, filed on Dec. 22, 2004. CA 2031433 A1 6, 1991 (Continued) (51) Int. C. C07C 309/19 (2006.01) OTHER PUBLICATIONS A6 IK3I/85 (2006.01) (52) U.S. C. Abderhalden et al. (CAPLUS Abstract of Fermentforschung (1930), USPC ...... 562/44; 514/576 12, 180-222).* Wermuth, The Practice of Medicinal Chemsitry, 2d ed. (2003), 768 (58) Field of Classification Search pages. Chapters 9-10 provided.* USPC ...... 562/44; 514/576 IUPAC. Compendium of Chemical Terminology, 2nded. (The “Gold See application file for complete search history. Book”). Compiled by A. D. McNaught and A. Wilkinson. Blackwell Scientific Publications, Oxford (1997). "Amino-acid residue” defi (56) References Cited nition provided.* Cerovsky et al., 1994, Int. J. Peptide & Protein Res., 44(5), 466-471. U.S. PATENT DOCUMENTS De Bont et al., 1996, Bioorg & Med. Chem. 6(24), 3035-3040. 2,376,911 A 5, 1945 Graenacher et al. Higashiura et al., 1992, J. Chem. Res., Synopsis, (8), 250-251. 2,531,468 A 1 1/1950 Reynolds et al. Hollowood, C.J., 2003, Org. & Biomolec. Chem., 1(10), 1664-1675. 3,218,352 A 11, 1965 Freifelder et al. 3,236,881 A 2f1966 Distler et al. (Continued) 3,453,309 A 7, 1969 Westland 3,580,936 A 5, 1971 Patchett et al. 3,658,966 A 4, 1972 Tsunoo et al. Primary Examiner — Robert Havlin 3,872,125 A 3, 1975 Houlihan et al. (74) Attorney, Agent, or Firm — McCarter & English, LLP: 3,920,833. A 11/1975 Cook et al. Danielle L. Herritt 4,085,134 A 4, 1978 Redmore et al. 4,187,245 A 2f1980 Redmore et al. 4,199,601 A 4, 1980 Durlach (57) ABSTRACT 4.255.448 A 3, 1981 Fariello 4,267,194 A 5, 1981 Durlach 4,271,189 A 6, 1981 Durlach Methods, compounds, pharmaceutical compositions and kits 4,355,043 A 10, 1982 Durlach are described for treating or preventing amyloid-related dis 4,386,081 A 5/1983 Helgstrand et al. CaSC. 4,448,779 A 5, 1984 Blanchard et al. 4,521,619 A 6/1985 Kaplan 4,522,811 A 6/1985 Eppstein et al. 17 Claims, No Drawings US 8,642.801 B2 Page 2

(56) References Cited 2005/0031651 A1 2/2005 Gervais et al. 2005, OO38000 A1 2/2005 Kong et al. U.S. PATENT DOCUMENTS 2005/0142191 A1 6/2005 Legore 2005/0215562 A1 9/2005 Tremblay et al. 5,383,988 A 1/1995 Herrmann et al. 2006, OO14752 A1 1/2006 Weaver et al. 5,385,915 A 1/1995 Buxbaum et al. 2006, O116347 A1 6/2006 Kisilevsky et al. 5,389,623 A 2, 1995 Bodor 2006/0135479 A1 6/2006 SZarek et al. 5,399,331 A 3/1995 Loughrey et al. 2006.01670.57 A1 7/2006 Kong et al. 5,405,834 A 4, 1995 Bundgaard et al. 2006.0167095 A1 7/2006 Kisilevsky et al. 5,413,996 A 5/1995 Bodor 2006/0183800 A1 8/2006 Kong et al. 5,416,016 A 5, 1995 Low et al. 2006/0223855 A1 10/2006 Kong et al. 5.430,052 A 7/1995 Higashiura et al. 2006/0252829 A1 11/2006 Garceau et al. 5.434,137 A 7/1995 Black 2007/0010573 A1 1/2007 Kong et al. 5,442,043 A 8/1995 Fukuta et al. 2007, OO15737 A1 1/2007 Clark et al. 5,463,092 A 10, 1995 Hostetler et al. 2007/0021483 A1 1/2007 Chalifour et al. 5,466,683 A 1 1/1995 Sterling et al. 2007/0078O82 A1 4/2007 Kisilevsky et al. 5,525,727 A 6/1996 Bodor 2007/0265334 A1 1 1/2007 Kisilevsky et al. 5,527,527 A 6, 1996 Friden 2008.0004244 A1 1/2008 SZarek et al. 5,622,934 A 4/1997 Fricket al. 2008, OO15180 A1 1/2008 Kong et al. 5,643,562 A 7/1997 Kisilevsky et al. 2008.OO27097 A1 1/2008 Kong et al. 5,668,117 A 9/1997 Shapiro 2008, OO38192 A1 2/2008 Gervais 5,672,683 A 9, 1997 Friden et al. 2010/O190753 A1 7/2010 Kong et al. 5,728,375 A 3/1998 Kisilevsky et al. 5,776,970 A 7, 1998 Shechter et al. FOREIGN PATENT DOCUMENTS 5,780,510 A 7/1998 Carney 5,840,294 A 1 1/1998 Kisilevsky et al. 5,858,326. A 1/1999 Kisilevsky et al. f 2.99. At 12: 5,952,389 A 9/1999 Fogel DE 4004978 A1 8, 1991

5,989,592 A 1 1/1999 Collin EP O 115657 A1 8/1984

6,015,835 A 1/2000 Miyamoto et al. EP O293974 A1 12/1988 6,024,977 A 2/2000 Yatvin et al. EP 0309421 A2 3, 1989

6,265,437 B1 7/2001 Berthelon et al. EP O405834 A2 1, 1991

6,310,073 B1 10/2001 Kisilevsky et al. EP O464759 A2 1/1992

6,376,557 B1 4/2002 Zaveri EP 1090 625 A2 4? 2001 6,440,952 B2 8/2002 Szarek et al. EP 1298 125 A1 4, 2003

6,670,399 B2 12/2003 Green et al. JP 60-184564 A 9, 1985

6,989.401 B2 1/2006 Maeda et al. JP 2-78620 3, 1990 7,153,692 B2 12/2006 Campbell et al. JP 2-149341. A 6, 1990 7.244.764 B2 7/2007 Kong et al. JP 04-077423. A 3, 1992 7.253,306 B2 8/2007 Kong et al. JP 04-103530 A 4, 1992

7,311,893 B2 12/2007 Gervais et al. WO WO-88.09171 A1 12, 1988

7.598,269 B2 10/2009 Kong et al. WO WO-90,09789 A2 9, 1990 2001/0048941 A1 12/2001 Kisilevsky et al. WO WO-91/04.014 A1 4, 1991 2002fOO94335 A1 7, 2002 Chalifour et al. WO WO-91/14434 A1 10, 1991 2002/0115717 A1 8/2002 Gervais et al. WO WO-91/14438 A1 10, 1991

2003/0077833 A1 4/2003 Campbell et al. WO WO-93/10459 A1 5.1993

2003. O153584 A1 8, 2003 Weaver et al. WO WO-94/OO135 A1 12, 1993 2003. O194375 A1 10, 2003 Weaver et al. WO WO-94f0111.6 A1 1, 1994 2004/0006092 A1 1/2004 Chalifour et al. WO WO-94.01131 A1 1, 1994 2004/0096453 A1 5/2004 Kisilevsky et al. WO WO-94f02178 A1 2, 1994 2004/0113277 A1 6, 2004 Chiras et al. WO WO-94,03424 A1 2, 1994 2004/O138178 A1 7/2004 SZarek et al. WO WO-94,06450 A1 3, 1994 2004/0208875 A1 10/2004 Kisilevsky et al. WO WO-94,22437 A2 10, 1994 2004/022O138 A1 11/2004 Gervais et al. WO WO-94,276O2 A1 12, 1994 US 8,642.801 B2 Page 3

(56) References Cited Beilstein Registry No. 7023352, N-(1'-aza-cyclopenten-2'yl)-2- aminoethane Sulfonic acid, May 11, 1995: Campagna, Francesco et FOREIGN PATENT DOCUMENTS al, “Cyclic Amidine Analogues of Taurine and Homotaurine: Syn thesis and Effects on Rat Skeletal Muscle.” II Farmaco, vol. 49 WO WO-95.01096 A1 1, 1995 (10):653-658 (1994). WO WO-95/06477 A1 3, 1995 Beilstein Registry No. 8919306, Microxine, Jan. 24, 2002: Killday, WO WO-95, O7092 A1 3, 1995 WO WO-96.00537 A1 1, 1996 K. Brian et al., “MicroXine, a New cdc2 Kinase Inhibitor from the WO WO-96,04001 A1 2, 1996 Australian Marine Sponge Microxima Species,” J. Nat. Prod., vol. WO WO-96,04915 A1 2, 1996 64:525-526 (2001). WO WO-96,223.03 A1 T 1996 Beilstein Registry No. 6023409, N,N-bis(2-sulfonylethyl)-1- WO WO-96,281.87 A1 9, 1996 octanamine disodium salt, Jul. 22, 1993: Tomson, R. et al., “Prepara WO WO-96,37612 A1 11, 1996 tion and Properties of Surfactants of the Type of disodium Salts of WO WO-96,39129 A1 12, 1996 N,N-BIS(2-Sulfoethyl)-1-Alkanamins.” Appl. Chem. USSR, vol. WO WO-97.074O2 A1 2, 1997 57(9): 1885-1891 (1984). WO WO-97.09445 A1 3, 1997 Beilstein Registry No. 2272192, N-(2-Sulfo-ethyl)-benzamid, Jun. WO WO-97.09976 A2 3, 1997 29, 1989: Wood, J. Matthew etal, “Reactivity and the mechanisms of WO WO-97.14306 A1 4f1997 reactions of B-Sultams with nucleophiles,” J. Chem. Soc., vol. 2:938 WO WO-97.16.191 A1 5, 1997 WO WO-98, 11923 A1 3, 1998 946 (2002). WO WO-98.13046 A1 4f1998 Beilstein Registry No. 3948718, N-(Butyl-sulfonsaeure-(4))-DL WO WO-98.25938 A1 6, 1998 alanin, Mar. 19, 1991: Helferich, von Burckhardt etal, “Sultame von WO WO-99/06545 A2 2, 1999 Aminosauren.” Liebigs Ann. Chem... vol. 651:33-42 (1962). WO WO-99/08685 A1 2, 1999 Beilstein Registry No. 1712477, 2-leucylamino-ethanesulfonic acid. WO WO-99/38498 A1 8, 1999 Feb. 27, 1989: Abderhalden, Emil et al., “Weitere Studien über das WO WO-99/40909 A1 8, 1999 Wesen von Ferment-wirkungen, ausgeführt mit Fermenten der WO WO-00,06133 A2 2, 2000 Gruppe der Polypeptidasen.” Fermentforschung, vol. 12:183-223 WO WO-00, 56328 A1 9, 2000 (1930). WO WO-OO,57707 A1 10, 2000 Beilstein Registry No. 2972476, 3-Benzylamino-propan-1- WO WO-00,64420 A2 11/2000 Sulfonsaeure, Jul. 11, 1989: Dorn, Helmut et al. "Cyanathylierung WO WO-0071101 A2 11/2000 und Sulfopropylierung von Phenyl-, Benzyl- und Cyclohexyl WO WO-01/03680 A2 1, 2001 WO WO-01/30979 A1 5, 2001 hydrazin.” Z. Chem., vol. 7:151-152 (1967). WO WO-01 (39796 A2 6, 2001 Beilstein Registry No. 2434022, 4-Aethylamino-butan WO WO-O152903 A1 T 2001 Sulfonsaeure-(1), Jul. 5, 1989: Helferich, von Burckhardt et al. WO WO-01 66533 A1 9, 2001 “Alkylamino- und Arylaminoalkansulfonsäuren Sowie WO WO-01,85093 A2 11/2001 Arylaminobutansultame.” Liebigs Ann. Chem... vol. 647:37-40 WO WO-O2/O7781 A2 1, 2002 (1961). WO WO-O2/O96937 A2 12/2002 Beilstein Registry No. 5620601, H-O-Glu-Ser-Tau-OH, Feb. 12, WO WO-O2/O97116 A2 12/2002 1993: Ienaga, Kazuharu et al. “Simple Peptides. III. Syntheses and WO WO-2004/058258 A1 T 2004 Properties of Taurine-Oligopeptides Containing an Acidic O-Amino WO WO-2004,113277 A2 12/2004 Acid.” Chem. Pharm. Bull., vol. 36(8):2796-2801 (1988). WO WO-2004 113275 A2 12/2004 Beilstein Registry No. 2846394, B-Naphthylaminomethylsulfonsaeure, Jul. 11, 1989. OTHER PUBLICATIONS Beilstein Registry No. 3952462, 4-AZonia-6-phenyl-hexan-1- IUPAC Gold Book, Compend of Chem. Terminol., 1997, 2" ed. Sulfonat, Mar. 19, 1991: Allen, C.F.H. etal, "Sultones as Reagents for “amino acid residue'. Derivatizing Aliphatic Amines in Qualitative Organic Analysis.” Koo E. H. et al., 1999, PNAS, 96,9989-9990. Anal. Chem., vol. 37:156-158 (1965). K.A. Muirhead et al., 1984, Cytometry, 5(3), 268-274. Beilstein Registry No. 5568774, L-phenylalanyltaurine, Feb. 12, M. Shengelia et al. 1998 Bull. Georg. Acad. Sc., 158(2), 250-3. 1993: Ienaga, Kazuharu et al. “Simple Peptides. II. Syntheses and Suvorov N.N. et al., 1977, Trudy Instituta Moskovskii Khimiko Properties of Taurine-Dipeptides Containing Neutral O-Amino tekhnologicheskii imeni D.I. Mendeleeva, 94.9-19. Acid.” Chem. Pharm. Bull., vol. 36(1):70-77 (1988). Suvorov N.N. et al., 1973, Khimiya Geterotsiklicheskikh Soedineni, Berge, Stephen M. etal, “Pharmaceutical Salts.” Journal of Pharma (11), 1505-11. ceutical Sciences, vol. 66(1):1-19 (1977). Tilley, J. W. et al., 1980, J. Med. Chem. 23, 1387-1392. Bloeman, P.G.M. et al., “Adhesion molecules, a new target for Zeid & Ismail, 1973, Chemistry & Industry, 380. immunoliposome-mediated drug delivery.” FEBS, vol. 357:140-144 Ancsin, John B. etal, “The Heparin/Heparan Sulfate-binding Site on (1995). Apo-serum Amyloid A: Implications for the Therapeutic Interven Boismare, F. etal, "A Homotaurine Derivative Reduces the Voluntary tion of Amyloidosis.” The Journal of Biological Chemistry, vol. Intake of Ethanol by Rats: are Cerebral GABA Receptors Involved?” 274(11):7172-7181 (1999). Pharmacology Biochemistry & Behavior, vol. 21:787-789 (1984). Aprile, Carlo et al., “Cardiac and pleuropulmonary AL amyloid imag Briggs, Andrew D. et al., “Acyloxymethyl and 4-Acyloxybenzyl ing with technetium-99m labelled aprotinin.” European Journal of Diester Prodrugs of Phosphonoformate.” Tetrahedron, vol. 52(47): 14937-14950 (1996). Nuclear Medicine, vol. 22(12): 1393-1401 (1995). Briscoe, Page et al., “Delivery of Superoxide dismutase to pulmonary Axelrad, M.A. et al., “Further Characterization of Amyloid-Enhanc epithelium via pH-sensitive liposomes.” Am. J. Physiol., vol. ing Factor.” Laboratory Investigation, vol. 47(2): 139-146 (1982). 268:L374-L380 (1995). Baures, Paul W. et al., “Discovering Transthyretin Amyloid Fibril Brissette, Louise et al., “Differential Induction of the Serum Amyloid Inhibitors by Limited Screening.” Bioorganic & Medicinal Chemis A Gene Family in Response to an Inflammatory Agent and to try, vol. 6: 1389-1401 (1998). Amyloid-enhancing Factor.” The Journal of Biological Chemistry, Beilstein Registry No. 338.851 1, 2-dibenzylamino-ethanesulfonic vol. 264(32):19327-19332 (1989). acid, Feb. 15, 1990. Buée, L. et al., “Alzheimer's disease: binding of vascular and Beilstein Registry No. 4261672, 2 <<3-hydroxy-5- neuroblastoma heparan Sulfate proteoglycans to amyloid f protein (hydroxymethyl)-2-methyl-4-pyridyl-methylamino-ethane A4.” Advances in the Biosciences, vol. 87:217-218 (1993). Sulphonic acid, Jul. 20, 1992: Iskander, MN et al. “Tansition-state Cai, Xiao-Dan et al., “Release of Excess Amyloid f Protein from a analogues as inhibitors for GABA-aminotransferase.” Eur: J. Med. Mutant Amyloid f Protein Precursor.” Science, vol. 259:514-516 Chem., vol. 26: 129-136 (1991). (1993). US 8,642.801 B2 Page 4

(56) References Cited Grant, K.A. et al., “Reinforcing and Discriminative Stimulus Effects of Ca-Acetyl Homotaurine in Animals.” Pharmacology, Biochemis OTHER PUBLICATIONS try & Behavior, vol. 32:607-611 (1989). Hamazaki, Hideaki et al., "Calcium-dependent polymerization of Campagna, Francesco et al., “Cyclic Amidine Analogues of Taurine human serum amyloid P component is inhibited by heparin and and Homotaurine: Synthesis and Effects on Rat Skeletal Muscle.” II dextran sulfate.” Biochimica et Biophysica Acta, vol. 998:231-235 Farmaco, vol. 49(10):653-658 (1994). (1989). Caughey, Byron et al. "Sulfated Polyanion Inhibition of Scrapie Hamazaki, Hideaki, "Ca-mediated Association of Human Serum Associated PrP Accumulation in Cultured Cells,” Journal of Virol Amyloid P Component with Heparan Sulfate and Dermatan Sulfate.” ogy, vol. 67(2):643-650 (1993). The Journal of Biological Chemistry, vol. 262(4): 1456-1460 (1987). Caughey, B., "Scrapie associated PrP accumulation and its preven Hamilton, Ronald L. "Lewy Bodies in Alzheimer's Disease: A tion: insights from cell culture.” British Medical Bulletin, vol. Neuropathological Review of 145 Cases. Using C-Synuclein 49(4):860-872 (1993). Immunohistochemistry.” Brain Pathology, vol. 10:378-384 (2000). Caughey, B., "Protease-resistant PrP accumulation and scrapie agent Han, Hogyu et al. “The core Alzheimer's peptide NAC forms replication: a role for Sulphated glycosaminoglycans?” Biochemical amyloid fibrils which seed and are seeded by B-amyloid: is NAC a Society Transactions, 648th Meeting Belfast, vol. 22:163-167 (1994). common trigger or target in neurodegenerative disease?” Chemistry Caughey, Byron, "Scrapie-associated PrP accumulation and agent & Biology, vol. 2:163–169 (1995). replication: effects on Sulphated glycosaminoglycan analogues.” Hawkins, P.N., “Diagnosis and monitoring of amyloidosis.” Phil. Trans. R. Soc. Lond. B., vol. 343:399-404 (1994). Bailliere's Clinical Rheumatology, vol. 8(3):635-659 (1994). Caughey, Byron et al., “Binding of the Protease-Sensitive Form of Hutchings, R. et al. “The Effect of Excitotoxin Antagonists on Prion Protein PrP to Sulfated Glycosaminoglycan and Congo Red.” Ibotenic Acid-Induced Alteration of APP MRNA Hippocampal Virology, vol. 68(4):2135-2141 (1994). Expression.” J. Pharmacy and Pharmacology, vol. 47(12B): 1131 Chabenat, C. et al., “Physicochemical, Pharmacological and (1995). Pharmacokinetic Study of a New GABAergic Compound, Calcium Ismail, Ibrahim Imam, “Reactions with Sultones II.” Afinidad L., vol. Acetylhomotaurinate.” Meth and Find Exptl Clin Pharmacol., vol. 446:256-258 (1993). 10(5):311-317 (1988). Ismail, Ibrahim Imam, “Reactions with Sultones and Sultams.” J. Chishti, M. Azhar et al. “Early-onset Amyloid Deposition and Cog Serb. Chem. Soc., vol. 57(7):415-420 (1992). nitive Deficits in Transgenic Mice Expressing a Double Mutant Form Iwai, Akihiko, “Properties of NACPo-synuclein and its role in of Amyloid Precursor Protein 695.” The Journal of Biological Chem Alzheimer's disease.” Biochimica et Biophysica Acta, vol. 1502:95 istry, vol. 276(24):21562-21570 (2001). 109 (2000). Colon, Wilfredo et al., “Partial Denaturation of Transthyretin is Suf Iwai, Akihiko et al., “Non-AB Component of Alzheimer's Disease ficient for Amyloid Fibril Formation in Vitro.” Biochemistry, vol. Amyloid (NAC) Is Amyloidogenic.” Biochemistry, vol. 34:10139 31:8654-8660 (1992). 10145 (1995). Copani, A. et al., “Activation of Metabotropic Glutamate Receptors James, Guy L. et al., “Benzodiazepine Peptidomimetics: Potent Protects Cultured Neurons Against Apoptosis Induced by B-Amyloid Inhibitors of Ras Farnesylation in Animal Cells.” Science, vol. Peptide.” Molecular Pharmacology, vol. 47:890-897 (1995). 260:1937-1942 (1993). DeMattos, Ronald B. et al. "Brain to Plasma Amyloid-fi Efflux: a Kagan, D.Z. etal, "Congo Red Inhibition of Amylogenesis in Experi Measure of Brain Amyloid Burden in a Mouse Model of Alzheimer's mental Amyloidosis.” Problemy Tuberkuleza, vol. 40:72-74. (1974). Disease.” Science, vol. 295:2264-2267 (2002). Keinänen, Kari et al., “BioSynthetic lipid-tagging of antibodies.” Dow, Kimberly E. etal, “Effects of 4-deoxy-L-threo-pentose, a novel FEBS, vol. 346:123-126 (1994). carbohydrate, on neural cell proteoglycan synthesis and function.” Killion, Jerald J. et al. “Systemic Targeting of Liposome-Encapsu Biochimica et Biophysica Acta, vol. 1156:7-14 (1992). lated Immunomodulators to Macrophages for Treatment of Cancer Durbin, PHetal, “Evidence of Acamprosate Penetration into the Rat Metastasis.” Immunomethods, vol. 4:273-279 (1994). Brain.” Behavioural Pharmacology, vol. 6:620 (1995). Kisilevsky, R., “From arthritis to Alzheimer's disease: current con Ehlers, Bernhard et al., “Dextran Sulphate 500 Delays and Prevents cepts on the pathogenesis of amyloidosis.” Can. J. Physiol. Mouse Scrapie by Impairment of Agent Replication in Spleen.” The Pharmacol., vol. 65:1805-1815 (1987). Journal of General Virology, vol. 65:1325-1330 (1984). Kisilevsky, R. et al. “The Potential Significance of Sulphated Eisai America, Inc., “Aricept.” Copyright 1995-1996, Center Watch, Glycosaminoglycans as a Common Constituent of all Amyloids: or, Inc. http://www.centerwatch.com/patient/drugs/dru190.html. Perhaps Amyloid is not a Misnomer.” Medical Hypotheses, vol. Frasier, Paul E. et al. “Effects of Sulfate Ions on Alzheimer B/A4 26:231-236 (1988). Peptide Assemblies: Implications for Amyloid Fibril-Proteoglycan Kisilevsky, Robert, “Theme and Variations on a String of Amyloid.” Interactions,” Journal of Neurochemistry, vol. 59:1531-1540 (1992). Neurobiology of Aging, vol. 10:499-500 (1989). Fraser, Paul E. et al., “Fibril Formation by Primate, Rodent, and Kisilevsky, Robert, "Heparan Sulfate Proteoglycans in Dutch-Hemorrhagic Analogues of Alzheimer Amyloid f-Protein.” Amyloidogenesis: An Epiphenomenon, A Unique Factor, or the Tip Biochemistry, vol. 31: 10716-10723 (1992). of a More Fundamental Process.” Laboratory Investigation, vol. Fujii, Akira et al., “Probiotics. Antistaphylococcal and 63(5):589-591 (1990). Antifibrinolytic Activities of (D-Amino- and Kisilevsky, Robert, "A Critical Analysis of Postulated Pathogenetic co-Guanidinoalkanesulfonic Acids,” Journal of Medicinal Chemis Mechanisms in Amyloidogenesis.” Critical Reviews in Clinical try, vol. 18(5):502-505 (1975). Laboratory Services, vol. 29(1):59-82 (1992). Gervais, Francine, "Amyloid Those Deadly Fibrils.” Eur: Kisilevsky, Robert et al., “Arresting amyloidosis in vivo using Small Biopharm. Review, pp. 40-42 (2001). molecule anionic Sulphonates or Sulphates: implications for Gervais, Francine et al., “Proteoglycans and Amyloidogenic Proteins Alzheimer's disease.” Nature Medicine, vol. 1(2): 143-148 (1995). in Peripheral Amyloidosis.” Curr: Med. Chem.—Immun., Endoc. & Krogsgaard-Larsen, P. et al., “Novel (Gamma-Aminobutyric Acid) Metab. Agents, vol. 3:361-370 (2003). Agonists and Partial Agonists.” FIDIA Research Foundation Sympo Girault, J. et al., “Determination of calcium acetylhomotaurinate in sium Series (1991). human plasma and urine by combined gas chromatography-negative Lacoste, Anne-Marie et al. “Inhibition of D-Alanyl-D-Alanine ion chemical ionization mass spectrometry.” Journal of Chromatog Ligase in Different Bacterial Species by Amino Phosphonic Acids.” raphy, vol. 530(2):295-305 (1990). Current Microbiology, vol. 2: 113-117 (1979). Gorin, Boris I. et al., “A Novel Esterification Procedure Applied to Leveugle, B. et al., “Binding of heparan Sulfate glycosaminoglycan to Syntheis of Biologically Active Esters of Foscarnet.” Tetrahedron 3-amyloid peptide: inhibition of potentially therapeutic polysulfated Letters, vol. 38(16):2791-2794 (1997). compounds.” NeuroReport, vol. 5: 1389-1392 (1994). US 8,642.801 B2 Page 5

(56) References Cited Powell, D.S. et al., “Insulin and Polyionic Sulphonates Modify Human Islet Amyloid Polypeptide Fibril Aggregation in Vitro.” OTHER PUBLICATIONS Diabetologia, vol. 41 (Suppl. 1):656 (1998). Puchtler, H. et al. "Application of Thiazole Dyes to Amyloid under Lhuintre, Jean-Pierre et al., “Ability of Calcium Bis Acetyl Conditions of Direct Cotton Dyeing: Correlation of Histochemical Homotaurine, a Gaba Agonist, to Prevent RElapse in Weaned Alco and Chemical Data.” Histochemistry, vol. 77:431-445 (1983). holics.” The Lancet, vol. 1 (8436): 1014-1016 (1985). Ranade, Vasant V., “Drug Delivery Systems. 1. Site-Specific Drug Littleton, John, Acamprosate in alcohol dependence: how does it Delivery Using Liposomes as Carriers,” J. Clin. Pharmacol., vol. work?” Addiction, vol. 90:1179-1188 (1995). Lyon, A.W. et al., "Co-deposition of Basement Membrane Compo 29:685-694 (1989). nents during the Induction of Murine Splenic AA Amyloid.” Labo Rodier, Par P. Toffoliet N. etal, “Bis(acetamido-3 propanesulfonate ratory Investigation, vol. 64(6):785-790 (1991). 1) de Calcium (N-Acetylhomotaurinate de Calcium).” Acta Cryst., Madamba, Samuel G. et al., “Acamprosate (Calcium vol. C44: 1493-1494 (1988). Acetylhomotaurinate) Enhances the N-Methyl-D-Aspartate Compo Sadler, Isobel I. J. et al. “Sulphated compounds attenuate B-amyloid nent of Excitatory Neurotransmission in Rat Hippocampal CA1 Neu toxicity by inhibiting its association with cells.” NeuroReport, vol. rons InVitro.” Alcoholism. Clinical and Experimental Research, vol. 7:49-53 (1995). 20(4):651-658 (1996). St. Georgiev, Vassil et al., “Drug-Induced Modifications of the Malmusi, Luca et al., "1,2,3,4-Tetrahydroisoquinoline and Related Immune Response. 1. Substituted 1-Phenylisoquinolines,” Journal of Analogs of the Phenylalkylamine Designer Drug MDMA.” Med. Medicinal Chemistry, vol. 22(4):348-352 (1979). Chem. Res., vol. 6(6):412-426 (1996). Sass, Henning et al., “Relapse Prevention of Acamprosate.” Arch. Masliah, Eliezer et al., “Altered Presynaptic Protein NACP Is Asso Gen. Psychiatry, vol. 53:673-680 (1996). ciated with Plaque Formation and Neurodegeneration in Alzheimer's Schreier, Hans et al., “Targeting of Liposomes to Cells Expressing Disease.” American Journal of Pathology, vol. 148(1):201-210 CD4. Using Glycosylphosphatidylinositol-anchored gp120.” The (1996). Journal of Biological Chemistry, vol. 269(12):9090-9098 (1994). Masuda, Midori et al. “Effect of taurine on nonspecific protection Shue, Ho-Jane et al., “A Study of 3-Amino-N- against bacterial infection.” Database STN International, Chemical Hydroxypropanesulfonamide Derivatives as Potential GABA Abstracts Service. Accession No. 105:108004 (1985) (Abstract Agonists and Their Fragmentation to 3-AMinopropanesulfinic only). Acid.” Bioorganic & Medicinal Chemistry Letters, vol. 6(14): 1709 May, Patrick C. “Current progress on new therapies for Alzheimer's 1714 (1996). disease.” Drug Discovery Today, vol. 6(9):459-462 (2001). Silverman, Richard B., “Prodrugs and Drug Delivery Systems.” The McCubbin, William D. et al., “Circular-dichroism studies on two Organic Chemistry of Drug Design and Drug Action, Academic murine serum amyloid A proteins.” Biochem. J., vol. 256:775-783 Press, Inc. Chapter 8, pp. 352-401. (1988). Small, D.H. et al., “Association and Release of the Amyloid Protein Mimura, Tetsutaro et al. "A Novel Class of Enkephalinase Inhibitors Precursor of Alzheimer's Disease from Chick Brain Extracellular Containing a C-Terminal Sulfo Group.” J. Med. Chem... vol. 35:602 Matrix.” The Journal of Neuroscience, vol. 12(11):4143-4150 608 (1992). (1992). Morgan, Barry A. etal, "Approaches to the Discovery of Non-Peptide Snow, Alan D. et al. "Sulfated Glycosaminoglycans: A Common Ligands for Peptide Receptors and Peptidases.” Ann. Rep. Med. Constituent of All Amyloids?” Laboratory Investigation, vol. Chem. Virick F.J. (Ed.) pp. 243-253, Academic Press, San Diego, CA 56(1): 120-123 (1987). (1989). Snow, Alan D. et al. “Temporal Relationship between Mukaetova-Ladinska, E.B. et al., "o-Synuclein Inclusions in Glycosaminoglycan Accumulation and Amyloid Deposition during Alzheimer and Lewy Body Diseases,” Journal of Neuropathology Experimental Amyloidosis.” Laboratory Investigation, vol. and Experimental Neurology, vol. 59(5):408-417 (2000). 53(1):37-44 (1985). Nakada, Tsutomu et al., “Guanidinoethane sulfate: brain pH alkaline Snow, Alan D. et al. "Sulfated Glycosaminoglycans in Alzheimer's shifter.” NeuroReport, vol. 4:1035-1038 (1993). Disease.” Human Pathology, vol. 18(5):506-510 (1987). Narindrasorasak, Suree et al. “Characterization of High Affinity Snow, Alan David et al., "Characterization of Tissue and Plasma Binding between Laminin and Alzheimer's Disease Amyloid Precur Glycosaminoglycans during Experimental AA Amyloidosis and sor Proteins.” Laboratory Investigation; vol. 67(5):643-652 (1992). Acute Inflammation, Qualitative and Quantitative Analysis.” Labo Narindrasorasak, Suree etal, “High Affinity Interactions between the ratory Investigation, vol. 56(6):665-675 (1987). Alzheimer's B-Amyloid Precursor Proteins and the Basement Mem Snow, Alan David et al., “A Close Ultrastructural Relationship brane Form of Heparan Sulfate Proteoglycan.” The Journal of Bio between Sulfated Proteoglycans and AA Amyloid Fibrils.” Labora logical Chemistry, vol. 266(20): 12878-12883 (1991). tory Investigation, vol. 57(6):687-698 (1987). National Institute on Alcohol Abuse and Alcoholism No. 33 PH 366, Snow, A. D. et al. "Sulfated glycosaminoglycans in amyloid plaques Jul. 1996; "Alcohol Alerg.” http://pubs.niaaa.nih.gov/publications/ of prion diseases.” Acta Neuropathol., vol. 77:337-342 (1989). aa33.htm. Snow, Alan D. et al. "A Temporal and Ultrastructural Relationship Norén, Jan O. et al. “Synthesis of Esters and Phosphonoformic Acid Between Heparan Sulfate Proteoglycans and AA Amyloid in Experi and Their Antiherpes Activity,” Journal of Medicinal Chemistry, vol. mental Amyloidosis.” The Journal of Histochemistry and 26(2):264-270 (1982). Cytochemistry, vol. 39(10): 1321-1330 (1991). O'Brien, Timothy D. et al., “Human Islet Amyloid Polypeptide Strejan, G.H. etal. “Suppression of Chronic-Relapsing Experimental Expression in COS-1 Cells.” American Journal of Pathology, vol. Allergic Encephalomyelitis in Strain-13 Guinea Pigs by Administra 147(3):609-616 (1995). tion of Liposome-Associated Myelin Basic Protein.” Journal of Owais, Mohammad et al. “Chloroquine Encapsulated in Malaria Neuroimmunology, vol. 7:27-41 (1984). Infected Erythrocyte-Specific Antibody-Bearing Liposomes Effec Tape, C. et al., “Direct Evidence for Circulating apoSAA as the tively Controls Chloroquine-Resistant Plasmodium berghei Infec Precursor of Tissue AA Amyloid Deposits.” Scand. J. Immunol., vol. tions in Mice.” Antimicrobial Agents and Chemotherapy, vol. 28:317-324 (1988). 39(1): 180-184 (1995). Travis, John, "New Piece of Alzheimer's Puzzle,' Science, vol. Panula-Lehto, Elina et al., “Comparison of the Effects of 261:828-829 (1993). Intraventricular Taurine, GABA and Homotaurine on Serum Ueda, Kenji et al., “Molecular cloning of cDNA encoding an unrec Prolactin Levels in Male Rats.” Pharmacology and Toxicology, vol. ognized component of amyloid in Alzheimer disease.” Proc. Natl. 65: 152-156 (1989). Acad. Sci. USA, vol. 90:11282-11286 (1993). Pollack, Scott J. etal, "Sulfonated dyes attenuate the toxic effects of Umezawa, F. etal, "Liposome Targeting to Mouse Brain: Mannose as 3-amyloid in a structure-specific fashion. Neuroscience Letters, vol. a Recognition Marker,” Biochemical and Biophysical Research Com 197:211-214 (1995). munications, vol. 153(3):1038-1044 (1988). US 8,642.801 B2 Page 6

(56) References Cited Zeid, I.F. et al., “Synthesis of halogenated Sultams II.” Afinidad, vol. 434:252-254 (1991). OTHER PUBLICATIONS Zeid, Ibrahim et al., “Synthesis von N-Substituirten Aminosulfonsären, II.” Liebigs Ann. Chem., pp. 671-689 (1974). Westermark, P., Islet Pathology of Non-Insulin-dependent Diabetes Silverman, R.B., (the Org. Chem. of Drug Design and Drug Action, Mellitus (NIDDM), Diabetic Medicine, vol. 13:S46-S48 (1996). Academic Press, Inc.: San Diego, 1992 pp. 4-51). Whitworth, A. et al., “Is Acamprosate an Effective Treatment for Alcohol Dependence?” The Lancet, vol. 347: 1438-1442 (1996). U.S. Appl. No. 10/639,609, dated Feb. 6, 2008. Wong, S. et al., “Influence of Sulphate Ions on the Structure of AA The Merck Index.an Encyclopedia of Chemicals, Drugs and Amyloid Fibrils.” Scand. J. Immunol., vol. 32:225-232 (1990). Biologicals, p. 883, 1989. Wood, Stephen J. etal. “Selective Inhibition of AB Fibil Formation.” CAS Registry No. 53329-38-7; Nov. 16, 1984. The Journal of Biological Chemistry, vol. 271 (8):4086-4092 (1996). CAS Registry No. 475060-41-4; Dec. 4, 2002. Yoshimoto, Makoto et al. “NACP, the precursor protein of the non CAS Registry No. 1146526-01-3; May 13, 2009. amyloid 3/A4 protein (A3) component of Alzheimer disease CAS Registry No. 1146526-10-4; May 13, 2009. amyloid, binds Afs and stimulates Afs aggregation.” Proc. Natl. Acad. CAS Registry No. 1146526-11-5; May 13, 2009. Sci. USA, vol. 92.9141-9145 (1995). CAS Registry No. 392716-04-0; Feb. 15, 2002. Young, I.D. et al. “The ultrastructural localization of sulfated CAS Registry No. 211613-79-5, Sep. 23, 1998. proteoglycans is identical in the amyloids of Alzheimer's disease and CAS Registry No. 141593-98-8; May 29, 1992. AA, AL, senile cardiac and medullary carcinoma-associated CAS Registry No. 89344-03-6, Nov. 16, 1984. amyloidosis.” Acta Neuropathol, vol. 78:202-209 (1989). CAS Registry No. 89343-99-7; Nov. 16, 1984. Young, Iain D. et al. “Localization of the Basement Membrane Database Caplus Online: Chemical Abstracts Service, Columbus, Heparan Sulfate Proteoglycan in Islet Amyloid Deposits in Type II Ohio, US; Evangelisti, F. etal. “New Synthetic Sweetener. Prepara Diabetes Mellitus.” Arch Pathol Lab Med., vol. 116:951-954 (1992). tion of Terpenoid Sulfamates' Rivista Della Societa Italiana di EP 00202287.9 Communication mailed Jul. 24, 2002 including Par Scienza dell'Alimentazione.9(6) 435-6 (1980). Database accession tial European Search Report (8 pages). No. 1981:515755. EP 00202287.9 Communication mailed Feb. 11, 2003, including Partial European Search Report (15 pages). Database Caplus Online: Chemical Abstracts Service, Columbus, International Search Report for Application No. PCT/IB2004/ Ohio, US; Spillane, William J. et al. "Semi-Quantitative and quanti 002375, dated Jul. 13, 2005. tative structure-taste relationships for carbo- and hetero-sulfamate Smith et al., 2002, CAS: 138: 12504. (RNHSO-) Sweeteners' J. Chem. Soc., Perkin Transcriptions 2: United States Office Action for U.S. Appl. No. 10/639,609, dated Physical Organic Chemistry (1972-1999), (7), 741-6, (1989). Data Feb. 6, 2008. base accession No. 1989:573.193. International Search Report for Application No. PCT/IB2004/ Database Caplus Online: Chemical Abstracts Service, Columbus, 002337, dated Jul 13, 2005. Ohio, US; Oehme, Guenther et al. “Preparation of Chiral Water Allen, et al., (CAPLUS abstract of Anal. Chem. (1965), 37(1): 156-8; Soluble Sulfobetaines or Ammoniumalkane Sulfonates by Ring Accession No. 1965:77985). Opening of Chiral Amines with Aliphatic Sultones' Akademie der CAplus 87: 172883, Laboratoires Lucien, Compositions with com Wissenschaften der DDR, Germany. Feb. 21, 1991. Database acces bined analgesic, myorelaxing, anxiolytic adn antiinflammatory activ Sion No. 1991:470920 & DD 287256. ity, (1997). Database Caplus Online: Chemical Abstracts Service, Columbus, CAplus 138:12504. Smith, Jack V. "Method for assaying Ohio, US; Kirmse, Wolfgang et al. “Deamination Reactions. 36. biomolecules and other constituents using indicator conjugates with Decomposition of Methynorborane-2-diazonium ions' Chemische Synthetic nucleounits in lateral flow, liquid, and dry chemistry tech Berichte, 114(5) 1793-808 (1981). Database accession No. niques.” (2002). 1981:46.1300. Erman, William F. et al., “Syntheses and Facile Cleavage of Five Dubowchik G. M. et al. "A Lipophilic Polyamino-Bolaamphiphile Membered Ring Sultams.” The Journal of Organic Chemistry, vol. Designed to Dissipate pH Gradients Across a Bilayer Membrane: 26(12):4841-4850 (1961). Synthesis and Protein Transport'Tetrahedron Letters, 37(36), 64.65 Ismail, I. Imam et al., “Reactions with Sultones.” Afinidad, vol. 434:239-241 (1991). 6468 (1996). Kametani, Tetsuji et al., “Syntheses of Homotaurine Related Com Database Caplus Online: Chemical Abstracts Service, Columbus, pounds (Studies on the Syntheses of Heterocyclic Compounds Part Ohio, US; De Nardo, M. et al. “Relations Between Chemical Con CCCLV).” Pharmacometrics, vol. 4(4):713-720 (1970). stituents and Sweetness. XV. Sulfamic and Sulfochloric Acids' La Combe, Edward et al., “A New Reaction of Allylic Sulfones.” Farmaco, Edizione Scientifica39(2), 125-32 (1984). Database acces Journal of the American Chemical Society, vol. 83(16):3457-3461 Sion No. 1984:137568. (1961). Watanabe, Fumihiko et al. “Synthesis of Metabolites of S-1452, an Thiel, Teresa et al., “New Zwitterionic butanesulfonic acids that Orally Active Thromboxane A2 Receptor Antagonist” Chem. Pharm. extend the alkaline range of four families of good buffers: Evaluation Bull. 39(11) 2842-2848 (1991). for use in biological systems.” J. Biochem. Biophys. Methods, vol. 37: 117-129 (1998). * cited by examiner US 8,642,801 B2 1. 2 METHODS AND COMPOSITIONS FOR term referring to a group of diverse but specific protein depos TREATING AMYLOID-RELATED DISEASES its (intracellular or extracellular) which are seen in a number of different diseases. Though diverse in their occurrence, all RELATED APPLICATIONS amyloid deposits have common morphologic properties, 5 This application is a continuation application of U.S. stain with specific dyes (e.g., Congo red), and have a charac patent application Ser. No. 1 1/316,694, filed Dec. 21, 2005, teristic red-green birefringent appearance in polarized light which, in turn, claims priority to U.S. Provisional Patent after staining. They also share common ultrastructural fea Application 60/638,819, filed on Dec. 22, 2004. U.S. patent tures and common X-ray diffraction and infrared spectra. application Ser. No. 1 1/316,694, filed Dec. 21, 2005 is also a 10 Amyloid-related diseases can either be restricted to one organ continuation-in-part of U.S. patent application Ser. No. or spread to several organs. The first instance is referred to as 10/871,514, filed Jun. 18, 2004, now U.S. Pat. No. 7,414,076, “localized amyloidosis” while the second is referred to as issued Aug. 19, 2008, which claims priority to U.S. Provi “systemic amyloidosis.” sional Patent Application No. 60/512,047, filed Oct. 17, 2003, Some amyloid diseases can be idiopathic, but most of these and U.S. Provisional Patent Application No. 60/480,906, filed Jun. 23, 2003, all entitled Methods and Compositions for 15 diseases appear as a complication of a previously existing Treating Amyloid-Related Diseases. This application is also disorder. For example, primary amyloidosis (AL amyloid) related to U.S. Provisional Application No. 60/638,636, filed can appear without any other pathology or can follow plasma Dec. 22, 2004 and U.S. patent application Ser. No. 10/871, cell dyscrasia or multiple myeloma. 365, filed Jun. 18, 2004, now U.S. Pat. No. 7,244,764, issued Secondary amyloidosis is usually seen associated with Jul. 17, 2007. 2O chronic infection (such as tuberculosis) or chronic inflamma This application is also related to U.S. Provisional Patent tion (such as rheumatoid arthritis). A familial form of second Application No. 60/512,017, filed Oct. 17, 2003, U.S. Provi ary amyloidosis is also seen in other types of familial amy sional Patent Application No. 60/480,918, filed Jun. 23, 2003, loidosis, e.g., Familial Mediterranean Fever (FMF). This and U.S. patent application Ser. No. 10/871,613, filed Jun. 18, familial type of amyloidosis is genetically inherited and is 2004, abandoned, all entitled Methods for Treating Protein as found in specific population groups. In both primary and Aggregation Disorders. secondary amyloidosis, deposits are found in several organs This application is related to U.S. Provisional Patent Appli and are thus considered systemic amyloid diseases. cation No. 60/512,116, filed Oct. 17, 2003, U.S. Provisional Patent Application No. 60/480,984, filed Jun. 23, 2003, and “Localized amyloidoses’ are those that tend to involve a U.S. application Ser. No. 10/871,549, filed Jun. 18, 2004, single organ system. Different amyloids are also character abandoned, all entitled Pharmaceutical Formulations of 30 ized by the type of protein present in the deposit. For example, Amyloid-Inhibiting Compounds. neurodegenerative diseases such as scrapie, bovine spongi This application is related to U.S. Provisional Patent Appli form encephalitis, Creutzfeldt-Jakob disease, and the like are cation No. 60/436,379, filed Dec. 24, 2002, U.S. Provisional characterized by the appearance and accumulation of a pro Patent Application No. 60/482,214, filed Jun. 23, 2003, tease-resistant form of a prion protein (referred to as AScror entitled Combination Therapy for the Treatment of Alzhe- 35 PrP-27) in the central nervous system. Similarly, Alzheimer's imer's Disease, U.S. patent application Ser. No. 10/746,138. disease, another neurodegenerative disorder, is characterized abandoned, filed Dec. 24, 2003, International Patent Appli by neuritic plaques and neurofibrillary tangles. In this case, cation No. PCT/CA2003/002011, filed Dec. 24, 2003, and the amyloid plaques found in the parenchyma and the blood U.S. patent application Ser. No. 10/871,537, filed Jun. 18, vessel is formed by the deposition of fibrillar AB amyloid 2004, pending, entitled Therapeutic Formulations for the 40 protein. Other diseases such as adult-onset diabetes (type II Treatment of Beta-Amyloid Related Diseases. diabetes) are characterized by the localized accumulation of This application is related to U.S. Provisional Patent Appli amyloid fibrils in the pancreas. Once these amyloids have cation No. 60/512,135, filed Oct. 17, 2003, U.S. Provisional formed, there is no known, widely accepted therapy or treat Patent Application No. 60/482,058, filed Jun. 23, 2003, both ment which significantly dissolves amyloid deposits in situ, entitled Synthetic Process for Preparing Compounds for as prevents further amyloid deposition or prevents the initiation Treating Amyloidosis, and U.S. patent application Ser. No. of amyloid deposition. 10/871,543, filed Jun. 18, 2004, now U.S. Pat. No. 7,253,306, Each amyloidogenic protein has the ability to undergo a issued Aug. 7, 2007, entitled Improved Pharmaceutical Drug Candidates and Method for Preparation Thereof. conformational change and to organize into B-sheets and This application is related to U.S. Provisional Patent Appli form insoluble fibrils which may be deposited extracellularly cation Ser. No. 60/512,018, filed on Oct. 17, 2003 and U.S. " or intracellularly. Each amyloidogenic protein, although dif Provisional Patent Application Ser. No. 60/480,928, filed on ferent in amino acid sequence, has the same property of Jun. 23, 2003, and U.S. patent application Ser. No. 10/871, forming fibrils and binding to other elements such as pro 512, filed Jun. 18, 2004, pending, all entitled Methods and teoglycan, amyloid P and complement component. More Compositions for Treating Amyloid- and Epileptogenesis over, each amyloidogenic protein has amino acid sequences Associated Diseases. 55 which, although different, show similarities such as regions This application is also related to Method for Treating with the ability to bind to the glycosaminoglycan (GAG) Amyloidosis, U.S. patent application Ser. No. 08/463,548, portion of proteoglycan (referred to as the GAG binding site) now U.S. Pat. No. 5,972,328. as well as other regions which promote B-sheet formation. The entire contents of each of these patent applications and Proteoglycans are macromolecules of various sizes and struc patents are hereby expressly incorporated herein by reference 60 tures that are distributed almost everywhere in the body. They including without limitation the specification, claims, and can be found in the intracellular compartment, on the Surface abstract, as well as any figures, tables, or drawings thereof. of cells, and as part of the extracellular matrix. The basic structure of all proteoglycans is comprised of a core protein BACKGROUND and at least one, but frequently more, polysaccharide chains 65 (GAGs) attached to the core protein. Many different GAGs Amyloidosis refers to a pathological condition character have been discovered including chondroitin Sulfate, dermatan ized by the presence of amyloid fibrils. Amyloid is a generic Sulfate, keratan Sulfate, heparin, and hyaluronan. US 8,642,801 B2 3 4 In specific cases, amyloid fibrils, once deposited, can described as diffuse or fibrillary. Both soluble oligomeric AB become toxic to the Surrounding cells. For example, the AB and fibrillar AB are also believed to be neurotoxic and inflam fibrils organized as senile plaques have been shown to be matory. associated with dead neuronal cells, dystrophic neurites, Another type of amyloidosis is cerebral amyloid angiopa astrocytosis, and microgliosis in patients with Alzheimer's 5 disease. When tested in vitro, oligomeric (soluble) as well as thy (CAA). CAA is the specific deposition of amyloid-f fibrillar AB peptide was shown to be capable of triggering an fibrils in the walls of leptomingeal and cortical arteries, arte activation process of microglia (brain macrophages), which rioles and veins. It is commonly associated with Alzheimer's would explain the presence of microgliosis and brain inflam disease, Down's syndrome and normal aging, as well as with mation found in the brain of patients with Alzheimer's dis 10 a variety of familial conditions related to stroke or dementia ease. Both oligomeric and fibrillar AB peptide can also induce (see Frangione et al., Amyloid. J. Protein Folding Disord. 8, neuronal cell death in vitro. See, e.g., M. P. Lambert, et al., Suppl. 1,36-42 (2001)). Proc. Natl. Acad. Sci. USA 95,6448-53 (1998). Presently available therapies for treatment of B-amyloid In another type of amyloidosis seen in patients with type II diseases are almost entirely symptomatic, providing only diabetes, the amyloidogenic protein IAPP, when organized in 15 temporary or partial clinical benefit. Although some pharma oligomeric forms or in fibrils, has been shown to induce ceutical agents have been described that offer partial Symp B-islet cell toxicity invitro. Hence, appearance of IAPP fibrils tomatic relief, no comprehensive pharmacological therapy is in the pancreas of type II diabetic patients contributes to the loss of the Bislet cells (Langerhans) and organ dysfunction currently available for the prevention or treatment of, for which can lead to insulinemia. example, Alzheimer's disease. Another type of amyloidosis is related to f microglobulin and is found in long-term hemodialysis patients. Patients SUMMARY OF THE INVENTION undergoing long term hemodialysis will develop f-micro globulin fibrils in the carpal tunnel and in the collagen rich The present invention relates to the use of certain com tissues in several joints. This causes severe pains, joint stiff 25 ness and Swelling. pounds in the treatment of amyloid-related diseases. In par Amyloidosis is also characteristic of Alzheimer's disease. ticular, the invention relates to a method of treating or pre Alzheimer's disease is a devastating disease of the brain that venting an amyloid-related disease in a Subject comprising results in progressive memory loss leading to dementia, administering to the Subject a therapeutic amount of a com physical disability, and death over a relatively long period of 30 pound of the invention. The invention also pertains to each of time. With the aging populations in developed countries, the the novel compounds of the invention as described herein. number of Alzheimer's patients is reaching epidemic propor Among the compounds for use in the invention are those tions. according to the following Formulae, such that, when admin People suffering from Alzheimer's disease develop a pro istered, amyloid fibril formation, organ specific dysfunction gressive dementia in adulthood, accompanied by three main 35 (e.g., neurodegeneration), or cellular toxicity is reduced or structural changes in the brain: diffuse loss of neurons in inhibited. multiple parts of the brain; accumulation of intracellular pro In one embodiment, the invention pertains, at least in part tein deposits termed neurofibrillary tangles; and accumula to compounds of Formula I: tion of extracellular protein deposits termed amyloid or senile plaques, Surrounded by misshapen nerve terminals (dystro 40 phic neurites) and activated microglia (microgliosis and (I) astrocytosis). A main constituent of these amyloid plaques is R2 the amyloid-fi peptide (AB), a 39-43 amino-acid protein that is produced through cleavage of the B-amyloid precursor R n 11 NS-YL protein (APP). Extensive research has been conducted on the 45 relevance of AB deposits in Alzheimer's disease, see, e.g., Selkoe, Trends in Cell Biology 8, 447-453 (1998). A? natu wherein: rally arises from the metabolic processing of the amyloid R" is a substituted or unsubstituted cycloalkyl, heterocy precursor protein (APP) in the endoplasmic reticulum clic, aryl, arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic (“ER), the Golgi apparatus, or the endosomal-lysosomal 50 pathway, and most is normally secreted as a 40 (AB1-40) or or tricyclic fused ring group, or a Substituted or unsubstituted 42 (A31-42) amino acid peptide (Selkoe, Annu. Rev. Cell C-Clo alkyl group: Biol. 10,373–403 (1994)). A role for AB as a primary cause for R is selected from a group consisting of hydrogen, alkyl, Alzheimer's disease is supported by the presence of extracel mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, lular AB deposits in senile plaques of Alzheimer's disease, the 55 thiazolyl, triazolyl, imidazolyl, benzothiazolyl, and ben increased production of AB in cells harboring mutant Alzhe Zoimidazolyl; imer's disease associated genes, e.g., amyloid precursor pro Y is SOX, OSOX", or SSO, X"; tein, presenilin I and presenilin II; and the toxicity of extra X" is hydrogen, a cationic group, or an ester-forming group cellular soluble (oligomeric) or fibrillar AB to cells in culture. (i.e., as in a prodrug, which are described elsewhere herein); See, e.g., Gervais, Eur: Biopharm. Review, 40-42 (Autumn 60 2001); May, DDT 6, 459-62 (2001). Although symptomatic and treatments exist for Alzheimer's disease, this disease cannot each of L' and L is independently a substituted or unsubsti be prevented or cured at this time. tuted C-C alkyl group or absent, or a pharmaceutically Alzheimer's disease is characterized by diffuse and neu acceptable salt thereof, provided that when R' is alkyl, L' is ritic plaques, cerebral angiopathy, and neurofibrillary tangles. 65 absent. Plaque and blood vessel amyloid is believed to be formed by In another embodiment, the invention pertains, at least in the deposition of insoluble AB amyloid protein, which may be part to compounds of Formula II: US 8,642,801 B2 5 6 genated alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, (II) aryl, cyano, thiazolyl, triazolyl, imidazolyl, tetrazolyl, ben R2 O Zothiazolyl, and benzoimidazolyl; and pharmaceutically acceptable salts and esters thereof, provided that said com . - (CH)-Y2 pound is not 3-(4-phenyl-1,2,3,6-tetrahydro-1-pyridyl)-1- R-L/ propanesulfonic acid. In yet another embodiment, the invention pertains at least wherein: in part to compounds of Formula IV: R" is a substituted or unsubstituted cyclic, bicyclic, tricy clic, or benzoheterocyclic group or a substituted or unsubsti 10 tuted C-C alkyl group; (IV) R is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, imidazolyl, R9 R8 R4a R5 benzothiazolyl, benzoimidazolyl, or linked to R' to form a R R5a heterocycle; 15 Y is SOX, OSO, X", or SSO, X"; R10 (CH)-N N-ch-i-A-R' X is hydrogen, a cationic group, or an ester forming moi 2 R7 Y K R6a O ety; R R R7a R6 m is 0 or 1; n is 1, 2, 3, or 4, L is substituted or unsubstituted C-C alkyl group or wherein: absent, or a pharmaceutically acceptable salt thereof, pro A is nitrogen or oxygen; vided that when R' is alkyl, L is absent. R'' is hydrogen, Salt-forming cation, ester forming group, In yet another embodiment, the invention pertains, at least in part to compounds of Formula III: -(CH2)-Q, or when A is nitrogen, A and R' taken together 25 may be the residue of a natural or unnatural amino acid or a salt or ester thereof; (III) Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia R4a R5 Zolyl, or benzoimidazolyl: R R5a O X is 0, 1, 2, 3, or 4: R3 30 n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; N-(CH)-S-A-RI R. R. R. R. R. R. R7, and R7 are each indepen R3a dently hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, R7 R6 O cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbo R7a R6a nyl, cyano, halogen, amino, tetrazolyl, R' and R taken 35 together, with the ring atoms they are attached to, form a wherein: double bond, or Rand R” taken together, with the ring atoms A is nitrogen or oxygen; they are attached to, form a double bond; R'' is hydrogen, Salt-forming cation, ester forming group, m is 0, 1, 2, 3, or 4: -(CH2)-Q, or when A is nitrogen, A and R' taken together R. R. R', R'', and R'' are independently selected from may be the residue of a natural or unnatural amino acid or a 40 a group of hydrogen, halogen, hydroxyl, alkyl, alkoxyl, halo salt or ester thereof; Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia genated alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, Zolyl, or benzoimidazolyl: aryl, cyano, thiazolyl, triazolyl, imidazolyl, tetrazolyl, ben X is 0, 1, 2, 3, or 4: Zothiazolyl, and benzoimidazolyl, and pharmaceutically n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 45 acceptable salts and esters thereof. R. R. R. R. R. R. R. R., R7 and R7 are each In another embodiment, the invention includes compounds independently hydrogen, alkyl, mercaptoalkyl, alkenyl, alky of Formula V: nyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycar bonyl, cyano, halogen, amino, tetrazolyl, or two R groups on adjacent ring atoms taken together with the ring atoms form a (V) double bond, provided that one of R. R. R. R. R. R. 50 R15 O R. R. R7 and R' is a moiety of Formula IIIa:

O (IIIa) 55 wherein: RB (CH2) A is nitrogen or oxygen; R'' is hydrogen, Salt-forming cation, ester forming group, -(CH2)-Q, or when A is nitrogen, A and R' taken together RC RE 60 may be the residue of a natural or unnatural amino acid or a RD salt or ester thereof; Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia wherein: Zolyl, or benzoimidazolyl: m is 0, 1, 2, 3, or 4: 65 X is 0, 1, 2, 3, or 4: R. R. R. R. and Rare independently selected from a n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; group of hydrogen, halogen, hydroxyl, alkyl, alkoxyl, halo aa is a natural or unnatural amino acid residue, US 8,642,801 B2 7 8 m is 0, 1, 2, or 3: X is 0, 1, 2, 3, or 4: R'' is hydrogen or protecting group; G is a direct bond or oxygen, nitrogen, or Sulfur, R" is hydrogen, alkyl or aryl, and pharmaceutically Z is 0, 1, 2, 3, 4, or 5: acceptable salts and prodrugs thereof. m is 0 or 1; In another embodiment, the invention includes compounds 5 R is selected from a group consisting of hydrogen, alkyl, of the Formula VI: mercaptoalkyl, alkenyl, alkynyl, aroyl, alkylcarbonyl, ami noalkylcarbonyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triaz olyl, imidazolyl, benzothiazolyl, and benzoimidazolyl: (VI) each R is independently selected from hydrogen, halo 10 gen, cyano, hydroxyl, alkoxy, thiol, amino, nitro, alkyl, aryl, f carbocyclic, or heterocyclic, and pharmaceutically accept R21 Yl O able salts thereof. \, --on In one embodiment, the compounds disclosed herein pre R201 R23 -c-)-cis--A-R R19 | vent or inhibit amyloid protein assembly into insoluble fibrils 15 which, in Vivo, are deposited in various organs, or it favors clearance of pre-formed deposits or slows deposition in wherein: patients already having deposits. In another embodiment, the n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; compound may also prevent the amyloid protein, in its A is oxygen or nitrogen; soluble, oligomeric form or in its fibrillar form, from binding R'' is hydrogen, salt-forming cation, ester forming group, or adhering to a cell Surface and causing cell damage or -(CH2)-Q, or when A is nitrogen, A and R' taken together toxicity. In yet another embodiment, the compound may may be the residue of a natural or unnatural amino acid or a block amyloid-induced cellular toxicity or macrophage acti salt or ester thereof; Vation. In another embodiment, the compound may block Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia amyloid-induced neurotoxicity or microglial activation. In Zolyl, or benzoimidazolyl: 25 another embodiment, the compound protects cells from amy X is 0, 1, 2, 3, or 4: loid induced cytotoxicity of B-islet cells. R" is hydrogen, alkyl or aryl; In another embodiment, the compound may enhance clear Y' is oxygen, sulfur, or nitrogen; ance from a specific organ, e.g., the brain or it decreases Y is carbon, nitrogen, or oxygen; concentration of the amyloid protein in Such a way that amy R" is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, 30 loid fibril formation is prevented in the targeted organ. alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetra The compounds of the invention may be administered Zolyl, imidazolyl, benzothiazolyl, or benzoimidazolyl: therapeutically or prophylactically to treat diseases associ R’ is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, ated with amyloid fibril formation, aggregation or deposition. cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imi The compounds of the invention may act to ameliorate the dazolyl, benzothiazolyl, benzoimidazolyl, or absent if Y is 35 course of an amyloid related disease using any of the follow OXygen; ing mechanisms (this list is meant to be illustrative and not R’ is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, limiting): slowing the rate of amyloid fibril formation or cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imi deposition; lessening the degree of amyloid deposition; dazolyl, benzothiazolyl, benzoimidazolyl: or R’ is hydro inhibiting, reducing, or preventing amyloid fibril formation; gen, hydroxyl, alkoxy or aryloxy if Y is nitrogen; or R’ is 40 inhibiting neurodegeneration or cellular toxicity induced by absent if Y is oxygen or sulfur, or RandR may be linked amyloid; inhibiting amyloid induced inflammation; enhanc to form a cyclic moiety if Y is nitrogen; ing the clearance of amyloid; or favoring the degradation of R’ is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, amyloid protein prior to its organization in fibrils. alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetra The compounds of the invention may be administered Zolyl, imidazolyl, benzothiazolyl, or benzoimidazolyl, or 45 therapeutically or prophylactically to treat diseases associ absent if Y is nitrogen or oxygen; ated with amyloid-fi fibril formation, aggregation or deposi or pharmaceutically acceptable salts thereof. tion. The compounds of the invention may act to ameliorate In another embodiment, the invention includes compounds the course of an amyloid-3 related disease using any of the of Formula VII: following mechanisms (this list is meant to be illustrative and 50 not limiting): slowing the rate of amyloid-fibril formation or deposition; lessening the degree of amyloid-fi deposition; (VII) inhibiting, reducing, or preventing amyloid-fi fibril forma tion; inhibiting neurodegeneration or cellular toxicity induced by amyloid-f; inhibiting amyloid-f induced inflam / \ (CH3)-N-(CH2)-S-A-RI 55 mation; enhancing the clearance of amyloid-fi from the brain; or favoring the degradation of amyloid-fi protein prior to its (R-G-(– k | organization in fibrils. Therapeutic compounds of the invention may be effective wherein: in controlling amyloid-fi deposition either following their n is 2, 3, or 4: 60 entry into the brain (following penetration of the blood brain A is oxygen or nitrogen; bather) or from the periphery. When acting from the periph R'' is hydrogen, salt-forming cation, ester forming group, ery, a compound may alter the equilibrium of AB between the —(CH),-Q, or when A is nitrogen, A and R' taken together brain and the plasma so as to favor the exit of AB from the may be the residue of a natural or unnatural amino acid or a brain. It may also increase the catabolism of neuronal Afand salt or ester thereof; 65 change the rate of exit from the brain. An increase in the exit Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia of AB from the brain would result in a decrease in AB brain Zolyl, or benzoimidazolyl: and cerebral spinal fluid (CSF) concentration and therefore US 8,642,801 B2 10 favor a decrease in AB deposition. Alternatively, compounds inflammation (such as rheumatoid arthritis). A familial form that penetrate the brain could control deposition by acting of secondary amyloidosis is also seen in Familial Mediterra directly on brain. A? e.g., by maintaining it in a non-fibrillar nean Fever (FMF). B microglobulin-related (dialysis-re form, favoring its clearance from the brain, or by slowing lated) amyloidosis is found in long-term hemodialysis down APP processing. These compounds could also prevent patients. Patients undergoing long term hemodialysis will A? in the brain from interacting with the cell surface and develop B-microglobulin fibrils in the carpal tunnel and in therefore prevent neurotoxicity, neurodegeneration or the collagen rich tissues in several joints. This causes severe inflammation. They may also decrease AB production by pains, joint stiffness and Swelling. These deposits are due to activated microglia. The compounds may also increase deg the inability to maintain low levels of BM in plasma of radation by macrophages or neuronal cells. 10 dialyzed patients. Increased plasma concentrations of BM In one embodiment, the method is used to treat Alzhe protein will induce structural changes and may lead to the imer's disease (e.g., sporadic, familial, or early AD). The deposition of modified BM as insoluble fibrils in the joints. method can also be used prophylactically or therapeutically DETAILED DESCRIPTION OF THE INVENTION to treat other clinical occurrences of amyloid-3 deposition, 15 Such as in Down's syndrome individuals and in patients with cerebral amyloid angiopathy (“CAA') or hereditary cerebral The present invention relates to the use of compounds of hemorrhage. Formulae I, II, III, IV, V, VI, VII, or compounds otherwise In another embodiment, the method is used to treat mild described herein in the treatment of amyloid-related diseases. cognitive impairment. Mild Cognitive Impairment (MCI) For convenience, some definitions of terms referred to herein is a condition characterized by a state of mild but measurable are set forth below. impairment in thinking skills, which is not necessarily asso Amyloid-Related Diseases ciated with the presence of dementia. MCI frequently, but not AA (Reactive) Amyloidosis necessarily, precedes Alzheimer's disease. Generally, AA amyloidosis is a manifestation of a number Additionally, abnormal accumulation of APP and of amy 25 of diseases that provoke a sustained acute phase response. loid-fi protein in muscle fibers has been implicated in the Such diseases include chronic inflammatory disorders, pathology of sporadic inclusion body myositis (IBM) chronic local or systemic microbial infections, and malignant (Askanas, et al., Proc. Natl. Acad. Sci. USA 93, 1314-1319 neoplasms. The most common form of reactive or secondary (1996); Askanas, et al., Current Opinion in Rheumatology 7. (AA) amyloidosis is seen as the result of long-standing 486-496 (1995)). Accordingly, the compounds of the inven 30 inflammatory conditions. For example, patients with Rheu matoid Arthritis or Familial Mediterranean Fever (which is a tion can be used prophylactically or therapeutically in the genetic disease) can develop AA amyloidosis. The terms 'AA treatment of disorders in which amyloid-beta protein is amyloidosis' and “secondary (AA) amyloidosis' are used abnormally deposited at non-neurological locations, such as interchangeably. AA fibrils are generally composed of 8,000 treatment of IBM by delivery of the compounds to muscle 35 Dalton fragments (AA peptide or protein) formed by pro fibers. teolytic cleavage of serum amyloid A protein (ApoSAA), a Additionally, it has been shown that AB is associated with circulating apolipoprotein which is mainly synthesized in abnormal extracellular deposits, known as drusen, that accu hepatocytes in response to Such cytokines as IL-1, IL-6 and mulate along the basal Surface of the retinal pigmented epi TNF. Once secreted, ApoSAA is complexed with HDL. thelium in individuals with age-related macular degeneration 40 Deposition of AA fibrils can be widespread in the body, with (AMD). AMD is a cause of irreversible vision loss in older a preference for parenchymal organs. The kidneys are usually individuals. It is believed that AB deposition could be an a deposition site, and the liver and the spleen may also be important component of the local inflammatory events that affected. Deposition is also seen in the heart, gastrointestinal contribute to atrophy of the retinal pigmented epithelium, tract, and the skin. drusen biogenesis, and the pathogenesis of AMD (Johnson, et 45 Underlying diseases which can lead to the development of al., Proc. Natl. Acad. Sci. USA99(18), 11830-5 (2002)). AA amyloidosis include, but are not limited to inflammatory The present invention therefore relates to the use of com diseases, such as rheumatoid arthritis,juvenile chronic arthri pounds of Formulae I, II, III, IV, V, VI, VII, or otherwise tis, ankylosing spondylitis, psoriasis, psoriatic arthropathy, described herein in the prevention or treatment of amyloid Reiter's syndrome. Adult Stills disease, Behcet’s syndrome, related diseases, including, inter alia, Alzheimer's disease, 50 and Crohn's disease. AA deposits are also produced as a cerebral amyloid angiopathy, mild cognitive impairment, result of chronic microbial infections, such as leprosy, tuber inclusion body myositis, Down's syndrome, macular degen culosis, bronchiectasis, decubitus ulcers, chronic pyelone eration, as well as other types of amyloidosis like IAPP phritis, osteomyelitis, and Whipple's disease. Certain malig related amyloidosis (e.g., diabetes), primary (AL) amyloido nant neoplasms can also result in AA fibril amyloid deposits. sis, secondary (AA) amyloidosis and B microglobulin 55 These include Such conditions as Hodgkin’s lymphoma, renal related (dialysis-related) amyloidosis. carcinoma, carcinomas of gut, lung and urogenital tract, basal In Type II diabetes related amyloidosis (IAPP), the amy cell carcinoma, and hairy cell leukemia. Other underlying loidogenic protein IAPP induces B-islet cell toxicity when conditions that may be associated with AA amyloidosis are organized in oligomeric forms or in fibrils. Hence, appear Castleman's disease and Schnitzler's syndrome. ance of IAPP fibrils in the pancreas of type II diabetic patients 60 AL Amyloidoses (Primary Amyloidosis) contributes to the loss of the B islet cells (Langerhans) and AL amyloid deposition is generally associated with almost organ dysfunction which leads to insulinemia. Primary amy any dyscrasia of the B lymphocyte lineage, ranging from loidosis (AL amyloid) is usually found associated with malignancy of plasma cells (multiple myeloma) to benign plasma cell dyscrasia and multiple myeloma. It can also be monoclonal gammopathy. At times, the presence of amyloid found as an idiopathic disease. 65 deposits may be a primary indicator of the underlying dys Secondary (AA) amyloidosis is usually seen associated crasia. AL amyloidosis is also described in detail in Current with chronic infection (such as tuberculosis) or chronic Drug Targets, 2004, 5 159-171. US 8,642,801 B2 11 12 Fibrils of AL amyloid deposits are composed of mono Hereditary Systemic Amyloidoses clonal immunoglobulin light chains or fragments thereof. More specifically, the fragments are derived from the N-ter There are many forms of hereditary systemic amyloidoses. minal region of the light chain (kappa or lambda) and contain Although they are relatively rare conditions, adult onset of all or part of the variable (V) domain thereof. Deposits symptoms and their inheritance patterns (usually autosomal generally occur in the mesenchymal tissues, causing periph dominant) lead to persistence of Such disorders in the general eral and autonomic neuropathy, carpal tunnel syndrome, population. Generally, the syndromes are attributable to point macroglossia, restrictive cardiomyopathy, arthropathy of large joints, immune dyscrasias, myelomas, as well as occult mutations in the precursor protein leading to production of dyscrasias. However, it should be noted that almost any tis variant amyloidogenic peptides or proteins. Table 1 Summa Sue, particularly visceral organs such as the kidney, liver, 10 rizes the fibril composition of exemplary forms of these dis spleen and heart, may be involved. orders. TABLE 1 Fibril Composition of Exemplary Amyloid-Related Diseases Genetic Fibril Peptide/Protein Variant Clinical Syndrome ATTR protein from Transthyretin Metis 0, many Familial amyloid polyneuropathy (FAP), and fragments others (Mainly peripheral nerves) ATTR protein from Transthyretin Thr45, Ala80, Cardiac involvement predominant without and fragments Ser84, Met111, neuropathy, familial amyloid Ile122 polyneuropathy, senile systemic amyloidosis, Tenosynovium N-terminal fragment of Arg26 Familial amyloid polyneuropathy (FAP), Apollipoprotein A1 (apoAI) (mainly peripheral nerves) N-terminal fragment of Arg26, Ostertag0type, non-neuropathic Apoliproprotein A1 (AapoAI) Arg50, Arg (predominantly visceral involvement) 60, others Aapo AII from Apollipoprotein AII Familial amyloidosis Lysozyme (Alys) Ostertag-type, non-neuropathic (predominantly visceral involvement) Fibrogen alpha chain fragment Leu554, Cranial neuropathy with lattic corneal VaS26 dystrophy Gelsolin fragment (Agel) Asn187, Cranial neuropathy with lattice corneal Tyr187 dystrophy Cystatin C fragment (ACyS) Gué8 Hereditary cerebral hemorrhage (cerebral amyloid angiopathy)-Icelandic type B-amyloid protein (AB) derived Glinó93 Hereditary cerebral hemorrhage (cerebral from Amyloid Precursor Protein (APP) amyloid angiopathy)-Dutch type B-amyloid protein (AB) derived Ile717, Phe717, Familial Alzheimer's Disease from Amyloid Precursor Protein (APP) Gly717 B-amyloid protein (AB) derived Glin 618 Alzheimer's disease, Down's syndrome, from Amyloid Precursor Protein (APP), hereditary cerebral hemorrhage with e.g., bPP 695 amyloidosis, Dutch type B-amyloid protein (AB) derived ASnó70, Familial Dementia-probably Alzheimer's from Amyloid Precursor Protein (APP) Leu671 Disease Prion Protein (PrP, APrPC) Leu102, Familial Creutzfeldt-Jakob disease: derived from Prp precursor Val167, Gerstmann-Sträussler-Scheinker syndrome protein (51-91 insert) Asn178, (hereditary spongiform encephalopathies, Lys200 prion diseases) AA derived from Serum amyloid Familial Mediterranean fever, predominant A protein (ApoSAA) renal involvement (autosomal recessive) AA derived from Serum amyloid Muckle-Well's syndrome, nephropathy, A protein (ApoSAA) deafness, urticaria, limb pain Unknown Cardiomyopathy with persistent atrial standstill Unknown Cutaneous deposits (bullous, papular, pustulodermal) AHamyloid protein, derived Ay I Myeloma associated amyloidosis from immunoglobulin heavy chain (gamma I) ACal amyloid protein from (Pro) Medullary carcinomas of the thyroid (pro)calcitonin calcitonin AANF amyloid protein from Isolated atrial amyloid atrial natriuretic factor Apro from Prolactin Prolactinomas Abri/ADan from ABripeptide British and Danish familial Dementia Data derived from Tan SY. Pepys MB. Amyloidosis. Histopathology, 25(5), 403-414 (November 1994), WHOIUIS Nomenclature Subcommittee, Nomenclature of Amyloid and Amyloidosis. Bulletin of the World Health Organisation 1993; 71: 10508; and Merlini et al., Clin Chem LabMed 2001;39(11): 1065-75. The data provided in Table 1 are exemplary and are not In general, any hereditary amyloid disorder can also occur intended to limit the scope of the invention. For example, sporadically, and both hereditary and sporadic forms of a more than 40 separate point mutations in the transthyretin 65 disease present with the same characteristics with regard to gene have been described, all of which give rise to clinically amyloid. For example, the most prevalent form of secondary similar forms of familial amyloid polyneuropathy. AA amyloidosis occurs sporadically, e.g. as a result of ongo US 8,642,801 B2 13 14 ing inflammation, and is not associated with Familial Medi majority being kappa, in some cases KIV. Bellotti et al., Jour terranean Fever. Thus general discussion relating to heredi nal of Structural Biology 13:280-89 (2000). Comparison of tary amyloid disorders below can also be applied to sporadic amyloidogenic and non-amyloidogenic heavy chains in amyloidoses. patients having AH amyloidosis has revealed missing and/or Transthyretin (TTR) is a 14 kiloDalton protein that is also altered components. Eulitz et al., Proc Natl Acad Sci USA sometimes referred to as prealbumin. It is produced by the 87:6542-46 (1990) (pathogenic heavy chain characterized by liver and choroid plexus, and it functions in transporting significantly lower molecular mass than non-amyloidogenic thyroid hormones and vitamin A. At least 50 variant forms of heavy chains); and Solomon et al. Am J Hemat 45(2) 171-6 the protein, each characterized by a singleamino acid change, (1994) (amyloidogenic heavy chain characterized as consist are responsible for various forms of familial amyloid poly 10 ing solely of the VH-D portion of the non-amyloidogenic neuropathy. For example, Substitution of proline for leucine at heavy chain). position 55 results in a particularly progressive form of neu Accordingly, potential methods of detecting and monitor ropathy; Substitution of methionine for leucine at position ing treatment of Subjects having or at risk of having AL, 111 resulted in a severe cardiopathy in Danish patients. LCDD, AH, and the like, include but are not limited to immu Amyloid deposits isolated from heart tissue of patients 15 noassaying plasma or urine for the presence or depressed with systemic amyloidosis have revealed that the deposits are deposition of amyloidogenic light or heavy chains, e.g., amy composed of a heterogeneous mixture of TTR and fragments loid W., amyloid K. amyloid KIV, amyloidy, or amyloid Y1. thereof, collectively referred to as ATTR, the full length Brain Amyloidosis sequences of which have been characterized. ATTR fibril The most frequent type of amyloid in the brain is composed components can be extracted from Such plaques and their primarily of AB peptide fibrils, resulting in dementia associ structure and sequence determined according to the methods ated with sporadic (non-hereditary) Alzheimer's disease. In known in the art (e.g., Gustaysson, A., et al., Laboratory fact, the incidence of sporadic Alzheimer's disease greatly Invest. 73: 703-708, 1995; Kametani, F., et al., Biochem. exceeds forms shown to be hereditary. Nevertheless, fibril Biophys. Res. Commun. 125: 622-628, 1984; Pras, M., et al., peptides forming plaques are very similarin both types. Brain PNAS80: 539-42, 1983). Persons having point mutations in 25 amyloidosis includes those diseases, conditions, pathologies, the molecule apolipoprotein A1 (e.g., Gly->Arg26; and other abnormalities of the structure or function of the Trp->Arg50; Leu->Arg60) exhibit a form of amyloidosis brain, including components thereof, in which the causative (“Ostertag type") characterized by deposits of the protein agent is amyloid. The area of the brainaffected in an amyloid apolipoprotein AI or fragments thereof (AApoAI). These related disease may be the Stroma including the vasculature or patients have low levels of high density lipoprotein (HDL) 30 the parenchyma including functional or anatomical regions, and present with a peripheral neuropathy or renal failure. or neurons themselves. A Subject need not have received a A mutation in the alpha chain of the enzyme lysozyme definitive diagnosis of a specifically recognized amyloid (e.g., Ile->Thr56 or Asp->His57) is the basis of another form related disease. The term "amyloid related disease' includes of Ostertag-type non-neuropathic hereditary amyloid brain amyloidosis. reported in English families. Here, fibrils of the mutant 35 Amyloid-fi peptide (AB) is a 39-43 amino acid peptide lysozyme protein (Alys) are deposited, and patients generally derived by proteolysis from a large protein known as Beta exhibit impaired renal function. This protein, unlike most of Amyloid Precursor Protein (“BAPP”). Mutations in BAPP the fibril-forming proteins described herein, is usually result in familial forms of Alzheimer's disease, Down's syn present in whole (unfragmented) form (Benson, M.D., et al. drome, cerebral amyloid angiopathy, and senile dementia, CIBA Fidn. Symp. 199: 104-131, 1996). 40 characterized by cerebral deposition of plaques composed of Immunoglobulin light chains tend to form aggregates in A? fibrils and other components, which are described in various morphologies, including fibrillar (e.g., AL amyloido further detail below. Known mutations in APP associated sis and AH amyloidosis), granular (e.g., light chain deposi with Alzheimer's disease occur proximate to the cleavage tion disease (LCDD), heavy chain deposition disease sites off or Y-secretase, or within AB. For example, position (HCDD), and light-heavy chain deposition disease (LH 45 717 is proximate to the site of gamma-secretase cleavage of CDD)), crystalline (e.g., Acquired Farconi's Syndrome), and APP in its processing to AB, and positions 670/671 are proxi microtubular (e.g., Cryoglobulinemia). AL and AH amyloi mate to the site of B-secretase cleavage. Mutations at any of dosis is indicated by the formation of insoluble fibrils of these residues may result in Alzheimer's disease, presumably immunoglobulin light chains and heavy chain, respectively, by causing an increase in the amount of the 42/43 amino acid and/or their fragments. In AL fibrils, lambda ( ) chains such 50 form of AB generated from APP. The familial form of Alzhe as wVI chains (W6 chains), are found in greater concentrations imer's disease represents only 10% of the subject population. than kappa (K) chains. WIII chains are also slightly elevated. Most occurrences of Alzheimer's disease are sporadic cases Merlini et al., Clin Chem Lab Med 39(11):1065-75 (2001). where APP and AB do not possess any mutation. The structure Heavy chain amyloidosis (AH) is generally characterized by and sequence of AB peptides of various lengths are well aggregates of gamma chain amyloid proteins of the IgG1 55 known in the art. Such peptides can be made according to subclass. Eulitz et al., Proc Natl AcadSci USA 87:6542-46 methods known in the art, or extracted from the brain accord (1990). ing to known methods (e.g., Glenner and Wong, Biochem. Comparison of amyloidogenic to non-amyloidogenic light Biophys. Res. Comm. 129,885-90 (1984); Glenner and Wong, chains has revealed that the former can include replacements Biochem. Biophys. Res. Comm. 122, 1131-35 (1984)). In or substitutions that appear to destabilize the folding of the 60 addition, various forms of the peptides are commercially protein and promote aggregation. AL and LCDD have been available. APP is expressed and constitutively catabolized in distinguished from other amyloid diseases due to their rela most cells. The dominant catabolic pathway appears to be tively small population monoclonal light chains, which are cleavage of APP within the AB sequence by an enzyme pro manufactured by neoplastic expansion of an antibody-pro visionally termed C-Secretase, leading to release of a soluble ducing B cell. AL aggregates typically are well-ordered fibrils 65 ectodomain fragment known as APPSC. This cleavage pre of lambda chains. LCDD aggregates are relatively amor cludes the formation of AB peptide. In contrast to this non phous aggregations of both kappa and lambda chains, with a amyloidogenic pathway, APP can also be cleaved by enzymes US 8,642,801 B2 15 16 known as B- and Y-secretase at the N- and C-termini of the AB, Unless otherwise specified, the term "amyloid” refers to respectively, followed by release of AB into the extracellular amyloidogenic proteins, peptides, or fragments thereof which space. To date, BACE has been identified as B-secretase can be soluble (e.g., monomeric or oligomeric) or insoluble (Vasser, et al., Science 286:735-741, 1999) and presenilins (e.g., having fibrillary structure or in amyloid plaque). See, have been implicated in Y-secretase activity (De Strooper, et e.g., M. P. Lambert, et al., Proc. Nat'l Acad. Sci. USA 95, al., Nature 391, 387-90 (1998)). The 39-43 amino acid AB 6448-53 (1998). “Amyloidosis” or "amyloid disease” or peptide is produced by sequential proteolytic cleavage of the "amyloid-related disease' refers to a pathological condition amyloid precursor protein (APP) by the B and Y secretases characterized by the presence of amyloid fibers. 'Amyloid” is enzyme. Although AB 40 is the predominant form produced, a generic term referring to a group of diverse but specific 5-7% of total AB exists as AB42 (Cappai et al., Int. J. Biochem. 10 protein deposits (intracellular or extracellular) which are seen Cell Biol. 31.885-89 (1999)). in a number of different diseases. Though diverse in their The length of the AB peptide appears to dramatically alter occurrence, all amyloid deposits have common morphologic its biochemical/biophysical properties. Specifically, the addi properties, stain with specific dyes (e.g., Congo red), and have tional two amino acids at the C-terminus of Af 42 are very a characteristic red-green birefringent appearance in polar hydrophobic, presumably increasing the propensity of AB 42 15 ized light after staining. They also share common ultrastruc to aggregate. For example, Jarrett, et al. demonstrated that tural features and common X-ray diffraction and infrared AB42 aggregates very rapidly in vitro compared to AB40, spectra. Suggesting that the longer forms of AB may be the important Gelsolin is a calcium binding protein that binds to frag pathological proteins that are involved in the initial seeding of ments and actin filaments. Mutations at position 187 (e.g., the neuritic plaques in Alzheimer's disease (Jarrett, et al., Asp->Asn; Asp->Tyr) of the protein result in a form of Biochemistry 32, 4693-97 (1993); Jarrett, et al., Ann. N.Y. hereditary systemic amyloidosis, usually found in patients Acad. Sci. 695, 144-48 (1993)). from Finland, as well as persons of Dutch or Japanese origin. This hypothesis has been further substantiated by the In afflicted individuals, fibrils formed from gelsolin frag recent analysis of the contributions of specific forms of AB in ments (Agel), usually consist of amino acids 173-243 (68kDa cases of genetic familial forms of Alzheimer's disease 25 carboxyterminal fragment) and are deposited in blood vessels (“FAD'). For example, the “London” mutant form of APP and basement membranes, resulting in corneal dystrophy and (APPV717I) linked to FAD selectively increases the produc cranial neuropathy which progresses to peripheral neuropa tion of AB 42/43 forms versus AB 40 (Suzuki, et al., Science thy, dystrophic skin changes and deposition in other organs. 264, 1336-40 (1994)) while the “Swedish' mutant form of (Kangas, H., et al. Human Mol. Genet. 5(9): 1237-1243, APP (APPK670N/M671L) increases levels of both AB40 and 30 1996). Other mutated proteins, such as mutant alpha chain of AB42/43 (Citron, et al., Nature 360, 672-674 (1992); Cai, et fibrinogen (AfibA) and mutant cyStatin C (Acys) also form al., Science 259, 514-16, (1993)). Also, it has been observed fibrils and produce characteristic hereditary disorders. AfibA that FAD-linked mutations in the Presenilin-1 (“PS1') or fibrils form deposits characteristic of a nonneuropathic Presenilin-2 (PS2) genes will lead to a selective increase in hereditary amyloid with renal disease; Acy's deposits are char Af342/43 production but not AB40 (Borchelt, et al., Neuron 35 acteristic of a hereditary cerebral amyloid angiopathy 17, 1005-13 (1996)). This finding was corroborated in trans reported in Iceland (Isselbacher, Harrison's Principles of genic mouse models expressing PS mutants that demonstrate Internal Medicine, McGraw-Hill, San Francisco, 1995; Ben a selective increase in brain A42 (Borchelt, op.cit.; Duff, et son, et al.). In at least Some cases, patients with cerebral al., Neurodegeneration 5(4), 293-98 (1996)). Thus the lead amyloid angiopathy (CAA) have been shown to have amyloid ing hypothesis regarding the etiology of Alzheimer's disease 40 fibrils containing a non-mutant form of cystatin C in conjunc is that an increase in AB42 brain concentration due to an tion with amyloid beta protein (Nagai, A., et al. Molec. Chem. increased production and release of AB42 or a decrease in Neuropathol. 33: 63-78, 1998). clearance (degradation or brain clearance) is a causative event Certain forms of prion disease are now considered to be in the disease pathology. heritable, accounting for up to 15% of cases, which were Multiple mutation sites in either AB or the APP gene have 45 previously thought to be predominantly infectious in nature. been identified and are clinically associated with either (Baldwin, et al., in Research Advances in Alzheimer's Dis dementia or cerebral hemorrhage. Exemplary CAA disorders ease and Related Disorders, John Wiley and Sons, New York, include, but are not limited to, hereditary cerebral hemor 1995). In hereditary and sporadic prion disorders, patients rhage with amyloidosis of Icelandic type (HCHWA-I); the develop plaques composed of abnormal isoforms of the nor Dutch variant of HCHWA (HCHWA-D; a mutation in AB); 50 mal prion protein (PrP). A predominant mutant isoform, the Flemish mutation of AB; the Arctic mutation of AB; the PrP, also referred to as AScr. differs from the normal cel Italian mutation of AB; the Iowa mutation of AB; familial lular protein in its resistance to protease degradation, insolu British dementia; and familial Danish dementia. CAA may bility after detergent extraction, deposition in secondary lyso also be sporadic. Somes, post-translational synthesis, and high B-pleated sheet As used herein, the terms “Bamyloid, "amyloid-f.” and 55 content. Genetic linkage has been established for at least five the like refer to amyloid 3 proteins or peptides, amyloid B mutations resulting in Creutzfeldt-Jacob disease (CJD), Ger precursor proteins or peptides, intermediates, and modifica stmann-Sträussler-Scheinker syndrome (GSS), and fatal tions and fragments thereof, unless otherwise specifically familial insomnia (FFI). (Baldwin, supra) Methods for indicated. In particular, AB” refers to any peptide produced extracting fibril peptides from Scrapie fibrils, determining by proteolytic processing of the APP gene product, especially 60 sequences and making Such peptides are known in the art peptides which are associated with amyloid pathologies, (e.g., Beekes, M., et al. J. Gen. Virol. 76: 2567-76, 1995). including AB1-39, AB1-40, AB1-41, AB1-42, and AB1-43. For example, one form of GSS has been linked to a PrP For convenience of nomenclature, AB1-42 may be referred mutation at codon 102, while telencephalic GSS segregates to herein as “AB(1-42)' or simply as AB42 or “AB (and with a mutation at codon 117. Mutations at codons 198 and likewise for any other amyloid peptides discussed herein). As 65 217 result in a form of GSS in which neuritic plaques char used herein, the terms “Bamyloid, "amyloid-f.” and AB acteristic of Alzheimer's disease contain PrP instead of AB are synonymous. peptide. Certain forms of familial CJD have been associated US 8,642,801 B2 17 18 with mutations at codons 200 and 210; mutations at codons Islet Amyloid Polypeptide and Diabetes 129 and 178 have been found in both familial CJD and FFI. Islet hyalinosis (amyloid deposition) was first described (Baldwin, Supra). over a century ago as the presence of fibrous protein aggre Cerebral Amyloidosis gates in the pancreas of patients with severe hyperglycemia 5 (Opie, E. L., J. Exp. Med. 5: 397-428, 1901). Today, islet Local deposition of amyloid is common in the brain, par amyloid, composed predominantly of islet amyloid polypep ticularly in elderly individuals. The most frequent type of tide (IAPP), or amylin, is a characteristic histopathological amyloid in the brain is composed primarily of AB peptide marker in over 90% of all cases of Type II diabetes (also fibrils, resulting in dementia or sporadic (non-hereditary) known as Non-Insulin Dependent Diabetes, or NIDDM). Alzheimer's disease. The most common occurrences of cere These fibrillar accumulations result from the aggregation of bral amyloidosis are sporadic and not familial For example, 10 the islet amyloid polypeptide (IAPP) or amylin, which is a 37 the incidence of sporadic Alzheimer's disease and sporadic amino acid peptide, derived from a larger precursor peptide, CAA greatly exceeds the incidence of familial AD and CAA. called pro-IAPP. Moreover, sporadic and familial forms of the disease cannot IAPP is co-secreted with insulin in response to B-cell secre be distinguished from each other (they differ only in the tagogues. This pathological feature is not associated with 15 insulin-dependent (Type I) diabetes and is a unifying charac presence or absence of an inherited genetic mutation); for teristic for the heterogeneous clinical phenotypes diagnosed example, the clinical symptoms and the amyloid plaques as NIDDM (Type II diabetes). formed in both sporadic and familial AD are very similar, if Longitudinal studies in cats and immunocytochemical not identical. investigations in monkeys have shown that a progressive Cerebral amyloid angiopathy (CAA) refers to the specific increase in islet amyloid is associated with a dramatic deposition of amyloid fibrils in the walls of leptomingeal and decrease in the population of insulin-secreting B-cells and cortical arteries, arterioles and veins. It is commonly associ increased severity of the disease. More recently, transgenic ated with Alzheimer's disease, Down's syndrome and normal studies have strengthened the relationship between IAPP aging, as well as with a variety of familial conditions related plaque formation and B-cell apoptosis and dysfunction, indi to stroke or dementia (see Frangione et al., Amyloid J. Pro 25 cating that amyloid deposition is a principal factor in increas tein Folding Disord. 8, Suppl. 1, 36-42 (2001)). CAA can ing severity of Type II diabetes. occur sporadically or be hereditary. IAPP has also been shown to induce B-islet cell toxicity in Senile Systemic Amyloidosis vitro, indicating that appearance of IAPP fibrils in the pan Amyloid deposition, either systemic or focal, increases creas of Type II or Type I diabetic patients (post-islet trans with age. For example, fibrils of wild type transthyretin 30 plantation) could contribute to the loss of the B-cell islets (TTR) are commonly found in the heart tissue of elderly (Langerhans) and organ dysfunction. In patients with Type II individuals. These may be asymptomatic, clinically silent, or diabetes, the accumulation of pancreatic IAPP leads to for may result in heart failure. Asymptomatic fibrillar focal mation of oligomeric IAPP, leading to a buildup of IAPP deposits may also occur in the brain (AB), corpora amylacea amyloid as insoluble fibrous deposits which eventually of the prostate (B. microglobulin), joints and seminal 35 destroys the insulin-producing B cells of the islet, resulting in vesicles. B cell depletion and failure (Westermark, P. Grimelius, L., Dialysis-Related Amyloidosis (DRA) Acta Path. Microbiol. Scand., sect. A. 81: 291-300, 1973; de Plaques composed off microglobulin (BM) fibrils com Koning, E. J. P. et al., Diabetologia 36: 378-384, 1993; and monly develop in patients receiving long term hemodialysis Lorenzo, A., et al., Nature 368: 756-760, 1994). Accumula or peritoneal dialysis. B. microglobulin is a 11.8 kiloDalton 40 tion of IAPP as fibrous deposits can also have an impact on the polypeptide and is the light chain of Class I MHC antigens, ratio of pro-IAPP to IAPP normally found in plasma by which are present on all nucleated cells. Under normal cir increasing this ratio due to the trapping of IAPP in deposits. cumstances, BM is usually distributed in the extracellular Reduction off cell mass can be manifested by hyperglycemia space unless there is an impaired renal function, in which case and insulinemia. This f-cell mass loss can lead to a need for BM is transported into tissues where it polymerizes to form 45 insulin therapy. amyloid fibrils. Failure of clearance such as in the case of Diseases caused by the death or malfunctioning of a par impaired renal function, leads to deposition in the carpal ticular type or types of cells can be treated by transplanting tunnel and other sites (primarily in collagen-rich tissues of the into the patient healthy cells of the relevant type of cell. This joints). Unlike other fibril proteins, BM molecules are not approach has been used for Type I diabetes patients. Often produced by cleavage of a longer precursor protein and are 50 pancreatic islet cells from a donor are cultured in vitro prior to generally present in unfragmented form in the fibrils. (Ben transplantation, to allow them to recover after the isolation son, Supra). Retention and accumulation of this amyloid pre procedure or to reduce their immunogenicity. However, in cursor has been shown to be the main pathogenic process many instances islet cell transplantation is unsuccessful, due underlying DRA. DRA is characterized by peripheral joint to death of the transplanted cells. One reason for this poor osteoarthropathy (e.g., joint stiffness, pain, Swelling, etc.). 55 Success rate is IAPP, which organizes into toxic oligomers. Isoforms offM, glycated fM, or polymers off M in tissue Toxic effects may result from intracellular and extracellular are the most amyloidogenic form (as opposed to native B2M). accumulation of fibril oligomers. The IAPP oligomers can Unlike other types of amyloidosis, B2M is confined largely to form fibrils and become toxic to the cells in vitro. In addition, osteoarticular sites. Visceral depositions are rare. Occasion IAPP fibrils are likely to continue to grow after the cells are ally, these deposits may involve blood vessels and other 60 transplanted and cause death or dysfunction of the cells. This important anatomic sites. may occur even when the cells are from a healthy donor and Despite improved dialysis methods for removal of BM, the patient receiving the transplant does not have a disease the majority of patients have plasmatic fM concentrations that is characterized by the presence of fibrils. For example, that remain dramatically higher than normal. These elevated compounds of the present invention may also be used in BM concentrations generally lead to Diabetes-Related Amy 65 preparing tissues or cells for transplantation according to the loidosis (DRA) and comorbidities that contribute to mortal methods described in International Patent Application (PCT) No. WO O1/OO368O. US 8,642,801 B2 19 20 The compounds of the invention may also stabilize the induced inflammation; enhancing the clearance of amyloid-f ratio of the concentrations of Pro-IAPP/IAPP pro-Insulin/ from the brain; or favoring greater catabolism of AB. Insulin and C-peptide levels. In addition, as biological mark Compounds of the invention may be effective in control ers of efficacy, the results of the different tests, such as the ling amyloid-fi deposition either following their entry into the arginine-insulin secretion test, the glucose tolerance test, brain (following penetration of the blood brain barrier) or insulin tolerance and sensitivity tests, could all be used as from the periphery. When acting from the periphery, a com markers of reduced B-cell mass and/or accumulation of amy pound may alter the equilibrium of AB between the brain and loid deposits. Such class of drugs could be used together with the plasma so as to favor the exit of AB from the brain. An other drugs targeting insulin resistance, hepatic glucose pro increase in the exit of A? from the brain would result in a 10 decrease in A brain concentration and therefore favor a duction, and insulin secretion. Such compounds might pre decrease in AB deposition. In addition, compounds that pen vent insulin therapy by preserving B-cell function and be etrate the brain may control deposition by acting directly on applicable to preserving islet transplants. brain AB, e.g., by maintaining it in a non-fibrillar form or Hormone-Derived Amyloidoses favoring its clearance from the brain. The compounds may Endocrine organs may harbor amyloid deposits, particu 15 slow down APP processing; may increase degradation of Af larly in aged individuals. Hormone-secreting tumors may fibrils by macrophages or by neuronal cells; or may decrease also contain hormone-derived amyloid plaques, the fibrils of AB production by activated microglia. These compounds which are made up of polypeptide hormones such as calcito could also prevent AB in the brain from interacting with the nin (medullary carcinoma of the thyroid), and atrial natri cell Surface and therefore prevent neurotoxicity, neurodegen uretic peptide (isolated atrial amyloidosis). Sequences and eration, or inflammation. structures of these proteins are well known in the art. In a preferred embodiment, the method is used to treat Miscellaneous Amyloidoses Alzheimer's disease (e.g., sporadic or familial AD). The There are a variety of other forms of amyloid disease that method can also be used prophylactically or therapeutically are normally manifest as localized deposits of amyloid. In to treat other clinical occurrences of amyloid-3 deposition, general, these diseases are probably the result of the localized 25 Such as in Down's syndrome individuals and in patients with production or lack of catabolism of specific fibril precursors cerebral amyloid angiopathy (“CAA'), hereditary cerebral or a predisposition of a particular tissue (such as the joint) for hemorrhage, or early Alzheimer's disease. fibril deposition. Examples of such idiopathic deposition In another embodiment, the method is used to treat mild include nodular AL amyloid, cutaneous amyloid, endocrine cognitive impairment. Mild Cognitive Impairment (MCI) amyloid, and tumor-related amyloid. Other amyloid related 30 is a condition characterized by a state of mild but measurable diseases include those described in Table 1, such as familial impairment in thinking skills, which is not necessarily asso amyloid polyneuropathy (FAP), senile systemic amyloidosis, ciated with the presence of dementia. MCI frequently, but not Tenosynovium, familial amyloidosis, Ostertag-type, non necessarily, precedes Alzheimer's disease. Additionally, neuropathic amyloidosis, cranial neuropathy, hereditary cere abnormal accumulation of APP and of amyloid-fi protein in bral hemorrhage, familial dementia, chronic dialysis, familial 35 muscle fibers has been implicated in the pathology of Spo Creutzfeldt-Jakob disease; Gerstmann-Sträussler-Scheinker radic inclusion body myositis (IBM) (Askanas, V., et al. syndrome, hereditary spongiform encephalopathies, prion (1996) Proc. Natl. Acad. Sci. USA93: 1314-1319; Askanas, V. diseases, familial Mediterranean fever, Muckle-Wells syn et al. (1995) Current Opinion in Rheumatology 7: 486-496). drome, nephropathy, deafness, urticaria, limb pain, cardi Accordingly, the compounds of the invention can be used omyopathy, cutaneous deposits, multiple myeloma, benign 40 prophylactically or therapeutically in the treatment of disor monoclonal gammopathy, maccoglobulinaemia, myeloma ders in which amyloid-beta protein is abnormally deposited at associated amyloidosis, medullary carcinomas of the thyroid, non-neurological locations. Such as treatment of IBM by isolated atrial amyloid, and diabetes. delivery of the compounds to muscle fibers. The compounds of the invention may be administered Additionally, it has been shown that AB is associated with therapeutically or prophylactically to treat diseases associ 45 abnormal extracellular deposits, known as drusen, that accu ated with amyloid fibril formation, aggregation or deposition, mulate along the basal Surface of the retinal pigmented epi regardless of the clinical setting. The compounds of the inven thelium in individuals with age-related macular degeneration tion may act to ameliorate the course of an amyloid related (ARMD). ARMD is a cause of irreversible vision loss in older disease using any of the following mechanisms, such as, for individuals. It is believed that AB deposition could be an example but not limited to: slowing the rate of amyloid fibril 50 important component of the local inflammatory events that formation or deposition; lessening the degree of amyloid contribute to atrophy of the retinal pigmented epithelium, deposition; inhibiting, reducing, or preventing amyloid fibril drusen biogenesis, and the pathogenesis of ARMD (Johnson, formation; inhibiting amyloid induced inflammation; et al., Proc. Natl. Acad. Sci. USA99(18), 11830-5 (2002)). enhancing the clearance of amyloid from, for example, the In another embodiment, the invention also relates to a brain; or protecting cells from amyloid induced (oligomers or 55 method of treating or preventing an amyloid-related disease fibrillar) toxicity. in a Subject (preferably a human) comprising administering to In an embodiment, the compounds of the invention may be the Subject a therapeutic amount of a compound according to administered therapeutically or prophylactically to treat dis the following Formulae or otherwise described herein, such eases associated with amyloid-fi fibril formation, aggregation that amyloid fibril formation or deposition, neurodegenera or deposition. The compounds of the invention may act to 60 tion, or cellular toxicity is reduced or inhibited. In another ameliorate the course of an amyloid-fi related disease using embodiment, the invention relates to a method of treating or any of the following mechanisms (this list is meant to be preventing an amyloid-related disease in a subject (preferably illustrative and not limiting): slowing the rate of amyloid-f a human) comprising administering to the Subject atherapeu fibril formation or deposition; lessening the degree of amy tic amount of a compound according to the following Formu loid-fi deposition; inhibiting, reducing, or preventing amy 65 lae or otherwise described herein, such that cognitive func loid-fi fibril formation; inhibiting neurodegeneration or cel tion is improved or stabilized or further deterioration in lular toxicity induced by amyloid-f; inhibiting amyloid-f cognitive function is prevented, slowed, or stopped in patients US 8,642,801 B2 21 22 with brain amyloidosis, e.g., Alzheimer's disease, Down's phatic chemistry as unimolecular (known as "Sy1') or bimo syndrome or cerebral amyloid angiopathy. These compounds lecular (“S2) reactions. Examples of nucleophiles include can also improve quality of daily living in these subjects. uncharged compounds such as amines, mercaptains, and alco The therapeutic compounds of the invention may treat hols, and charged groups such as alkoxides, thiolates, carban amyloidosis related to type II diabetes by, for example, sta 5 ions, and a variety of organic and inorganic anions. Illustra bilizing glycemia, preventing or reducing the loss of B cell tive anionic nucleophiles include, inter alia, simple anions mass, reducing or preventing hyperglycemia due to loss of B Such as azide, cyanide, thiocyanate, acetate, formate, or chlo cell mass, and modulating (e.g., increasing or stabilizing) roformate, and bisulfite. Organometallic reagents such as insulin production. The compounds of the invention may also organocuprates, organoZincs, organolithiums, Grignard stabilize the ratio of the concentrations of pro-IAPP/IAPP. 10 reagents, enolates, and acetylides, will under appropriate The therapeutic compounds of the invention may treat AA reaction conditions, be suitable nucleophiles. (secondary) amyloidosis and/or AL (primary) amyloidosis, Similarly, an “electrophile” means an atom, molecule, or by stabilizing renal function, decreasing proteinuria, increas ion able to accept an electron pair, particularly a pair of ing creatinine clearance (e.g., by at least 50% or greater or by electrons from a nucleophile, such as typically occurs during at least 100% or greater), by leading to remission of chronic 15 an electrophilic substitution reaction. In an electrophilic sub diarrhea or weight gain (e.g., 10% or greater), or by reducing stitution reaction, an electrophile binds to a substrate with the serum creatinine. Visceral amyloid content as determined, expulsion of another electrophile, e.g., the Substitution of a e.g., by SAP Scintigraphy may also be reduced. proton by another electrophile Such as a nitronium ion on an Compounds of the Invention aromatic Substrate (e.g., benzene). Electrophiles include The present invention relates, at least in part, to the use of cyclic compounds Such as epoxides, aziridines, episulfides, certain chemical compounds (and pharmaceutical formula cyclic Sulfates, carbonates, lactones, and lactams; and non tions thereof) in the prevention or treatment of amyloid-re cyclic electrophiles include Sulfates, Sulfonates (e.g., tosy lated diseases, including, inter alia, Alzheimer's disease, lates), chlorides, bromides, and iodides. Generally, an elec cerebral amyloid angiopathy, inclusion body myositis, trophile may be a saturated carbon atom (e.g., a methylene Down's syndrome, diabetes related amyloidosis, hemodialy 25 group) bonded to a leaving group; however, an electrophile sis-related amyloidosis (BM), primary amyloidosis (e.g., W may also be an unsaturated group, such as an aldehyde, or K chain-related), familial amyloid polyneuropathy (FAP), ketone, ester, or conjugated (C.B-unsaturated) analog thereof, senile systemic amyloidosis, familial amyloidosis, Ostertag which upon reaction with a nucleophile forms an adduct. type non-neuropathic amyloidosis, cranial neuropathy, The term “leaving group' generally refers to a group that is hereditary cerebral hemorrhage, familial dementia, chronic 30 readily displaced and Substituted by a nucleophile (e.g., an dialysis, familial Creutzfeldt-Jakob disease, Gerstmann amine, a thiol, an alcohol, or cyanide). Such leaving groups Sträussler-Scheinker syndrome, hereditary spongiform are well known and include carboxylates, N-hydroxysuccin encephalopathies, prion diseases, familial Mediterranean imide (“NHS), N-hydroxybenzotriazole, a halogen (fluo fever, Muckle-Wells syndrome, nephropathy, deafness, urti rine, chlorine, bromine, or iodine), alkoxides, and thioalkox caria, limb pain, cardiomyopathy, cutaneous deposits, mul 35 ides. A variety of Sulfur-based leaving groups are routinely tiple myeloma, benign monoclonal gammopathy, maccoglo used in synthetic chemistry, including alkane Sulfonyloxy bulinaemia, myeloma associated amyloidosis, medullary groups (e.g., C-C alkane Such as methane Sulfonyloxy, carcinomas of the thyroid, and isolated atrial amyloid. ethane Sulfonyloxy, propane Sulfonyloxy, and butane Sulfo The chemical structures herein are drawn according to the nyloxy groups) and the halogenated analogs (e.g., halogeno conventional standards known in the art. Thus, where an 40 (C-C alkane) sulfonyloxy groups, such as trifluoromethane atom, Such as a carbon atom, as drawn appears to have an Sulfonyloxy (i.e., triflate), 2.2.2-trichloroethane Sulfonyloxy, unsatisfied Valency, then that Valency is assumed to be satis 3,3,3-tribromopropane sulfonyloxy, and 4.4.4-trifluorobu fied by a hydrogen atom even though that hydrogen atom is tane Sulfonyloxy groups), as well as arylsulfonyloxy groups not necessarily explicitly drawn. The structures of some of the (e.g., Co-Co aryl optionally Substituted with 1 to 3 C-C, compounds of this invention include stereogenic carbon 45 alkyl groups, such as benzene Sulfonyloxy, C.-naphthylsulfo atoms. It is to be understood that isomers arising from Such nyloxy, B-naphthylsulfonyloxy, p-toluenesulfonyloxy (i.e., asymmetry (e.g., all enantiomers and diastereomers) are tosylates), 4-tert-butylbenzene Sulfonyloxy, mesitylene Sul included within the scope of this invention unless indicated fonyloxy, and 6-ethyl-O-naphthylsulfonyloxy groups). otherwise. That is, unless otherwise stipulated, any chiral “Activated esters' may be represented by the formula carbon center may be of either (R)- or (S)-stereochemistry. 50 -COL, where L is a leaving group, typical examples of which Such isomers can be obtained in substantially pure form by include N-hydroxysulfosuccinimidyl and N-hydroxysuccin classical separation techniques and by Stereochemically-con imidyl groups: aryloxy groups Substituted with electron trolled synthesis. Furthermore, alkenes can include either the withdrawing groups (e.g., p-nitro, pentafluoro, pentachloro, E- or Z-geometry, where appropriate. In addition, the com p-cyano, orp-trifluoromethyl); and carboxylic acids activated pounds of the present invention may exist in unsolvated as 55 by a carbodiimide to form an anhydride or mixed anhydride, well as solvated forms with acceptable solvents such as water, e.g., OCOR or OCNR'NHR, where R and R are THF, ethanol, and the like. In general, the solvated forms are independently C-C alkyl, Cs-Cs alkyl (e.g., cyclohexyl), considered equivalent to the unsolvated forms for the pur C-C perfluoroalkyl, or C-C alkoxy groups. An activated poses of the present invention. ester may be formed in situ or may be an isolable reagent. A “small molecule” refers to a compound that is not itself 60 Sulfo Succinimidyl esters, pentafluorothiophenol esters, and the product of gene transcription or translation (e.g., protein, sulfotetrafluorophenol are preferred activated esters. How RNA, or DNA) and preferably has a low molecular weight, ever, the ester leaving group may be, for example, Substituted e.g., less than about 2500 amu. or unsubstituted C-C alkyl (such as methyl, ethyl, propyl. In general, the term “nucleophile' is art-recognized to isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, or mean a chemical group having a reactive pair of electrons that 65 hexyl), or Substituted or unsubstituted C-C aryl or hetero reacts with a compound by displacing a leaving group (com cyclic groups, such as 2-fluoroethyl, 2-chloroethyl, 2-bromo monly another nucleophile), such as commonly occur in ali ethyl, 2,2-dibromoethyl, 2.2.2-trichloroethyl, 3-fluoropro US 8,642,801 B2 23 24 pyl, 4-chlorobutyl, methoxymethyl, 1,1-dimethyl-1- and to cycloalkyl groups having from 3 to 6 carbons in the methoxymethyl, ethoxymethyl, N-propoxymethyl, ring structure. Unless the number of carbons is otherwise isopropoxymethyl, N-butoxymethyl, tert-butoxymethyl, specified, “lower as in “lower aliphatic.” “lower alkyl.” 1-ethoxyethyl, 1-methyl-1-methoxyethyl, 1-(isopropoxy) “lower alkenyl, etc. as used herein means that the moiety has ethyl 3-methoxypropyl-4-methoxybutyl, fluoromethoxym at least one and less than about 8 carbon atoms. In certain ethyl, 2.2.2-trichloroethoxymethyl, bis(2-chloroethoxy)me embodiments, a straight-chain or branched-chain lower alkyl thyl, 3-fluoropropoxymethyl, 4-chlorobutoxyethyl, group has 6 or fewer carbon atoms in its backbone (e.g., dibromomethoxyethyl, 2-chloroethoxypropyl. fluo C-C for straight-chain, C-C for branched-chain), and romethoxybutyl, 2-methoxyethoxymethyl, ethoxymethoxy more preferably 4 or fewer. Likewise, preferred cycloalkyl ethyl, methoxyethoxypropyl, methoxyethoxybutyl, benzyl, 10 groups have from 3-8 carbonatoms in their ring structure, and phenethyl 3-phenylpropyl, 4-phenylbutyl, C.-naphthylm more preferably have 5 or 6 carbons in the ring structure. The ethyl, B-naphthylmethyl, diphenylmethyl, triphenylmethyl, term "C-C as in "C-C alkyl means alkyl groups con C.-naphthyldipheylmethyl, 9-anthrylmethyl, 4-methylbenzyl, taining 1 to 6 carbon atoms. 2,4,6-trimethylbenzyl, 3,4,5-trimethylbenzyl, 4-methoxy Moreover, unless otherwise specified the term alkyl benzyl, 4-methoxyphenyldiphenylmethyl 2-nitrobenzyl, 15 includes both “unsubstituted alkyls' and “substituted alkyls.” 4-nitrobenzyl, 4-chlorobenzyl, 4-bromobenzyl, 4-cyanoben the latter of which refers to alkyl groups having substituents Zyl, 4-cyanobenzyldiphenylmethyl, orbis(2-nitrophenyl)me replacing one or more hydrogens on one or more carbons of thyl groups. the hydrocarbon backbone. Such substituents may include, The term "electron-withdrawing group' is art-recognized for example, alkenyl, alkynyl, halogeno, hydroxyl, alkylcar and describes the ability of a substituent to attract valence bonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxy, ary electrons (e.g., pi-electrons) from neighboring atoms, e.g., loxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, the Substituent is more electronegative than neighboring alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, atoms, or it draws electrons to itself more than a hydrogen dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phos atom would at the same position. The Hammett sigma value phate, phosphonato, phosphinato, cyano, amino (including (O) is an accepted measure of a group's electron-donating and 25 alkyl amino, dialkylamino, arylamino, diarylamino, and withdrawing ability, especially the sigma para value (O). alkylarylamino), acylamino (including alkylcarbonylamino, See, e.g., “Advanced Organic Chemistry” by J. March, 5" arylcarbonylamino, carbamoyl and ureido), imino, Sulfhy Ed., John Wiley & Sons, Inc., New York, pp. 368-75 (2001). dryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfi The Hammett constant values are generally negative for elec nyl, Sulfonato, Sulfamoyl, Sulfonamido, nitro, trifluorom tron-donating groups (O -0.66 for NH2) and positive for 30 ethyl, cyano, azido, heterocyclic, alkylaryl, or aromatic electron-withdrawing groups (o, 0.78 for a nitro group), O, (including heteroaromatic) groups. indicating para substitution. Exemplary electron-withdraw An “arylalkyl group is an alkyl group substituted with an ing groups include nitro, acyl (ketone), formyl (aldehyde), aryl group (e.g., phenylmethyl (i.e., benzyl)). An “alkylary1 Sulfonyl, trifluoromethyl, halogeno (e.g., chloro and fluoro), moiety is an aryl group Substituted with an alkyl group (e.g., and cyano groups, among others. Conversely, an “electron 35 p-methylphenyl (i.e. p-tolyl)). The term “n-alkyl means a donating group' designates a Substituent that contributes straight-chain (i.e., unbranched) unsubstituted alkyl group. electrons more than hydrogen would if it occupied the same An “alkylene' group is a divalent analog of the corresponding position in the molecule. Examples include amino (including alkyl group. The terms “alkenyl and “alkynyl refer to unsat alkylamino and dialkylamino), aryl, alkoxy (including urated aliphatic groups analogous to alkyls, but which contain aralkoxy), aryloxy, mercapto and alkylthio, and hydroxyl 40 at least one double or triple carbon-carbon bond respectively. groups, among others. Suitable alkenyl and alkynyl groups include groups having 2 As used herein, “alkyl groups include Saturated hydrocar to about 12 carbonatoms, preferably from 2 to about 6 carbon bons having one or more carbon atoms, including straight atOmS. chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, The term 'aromatic group' or 'aryl group' includes unsat hexyl, heptyl, octyl, nonyl, decyl, etc.), cyclic alkyl groups (or 45 urated and aromatic cyclic hydrocarbons as well as unsatur “cycloalkyl or “alicyclic' or “carbocyclic' groups) (e.g., ated and aromatic heterocycles containing one or more rings. cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooc Aryl groups may also be fused or bridged with alicyclic or tyl, etc.), branched-chain alkyl groups (isopropyl, tert-butyl, heterocyclic rings that are not aromatic So as to form a poly sec-butyl, isobutyl, etc.), and alkyl-substituted alkyl groups cycle (e.g., tetralin). An “arylene' group is a divalent analog (e.g., alkyl-substituted cycloalkyl groups and cycloalkyl-sub 50 ofan aryl group. Aryl groups can also be fused orbridged with stituted alkyl groups). The term “aliphatic group' includes alicyclic or heterocyclic rings which are not aromatic So as to organic moieties characterized by Straight or branched form a polycycle (e.g., tetralin). chains, typically having between 1 and 22 carbon atoms. In The term "heterocyclic group' includes closed ring struc complex structures, the chains may be branched, bridged, or tures analogous to carbocyclic groups in which one or more of cross-linked. Aliphatic groups include alkyl groups, alkenyl 55 the carbon atoms in the ring is an element other than carbon, groups, and alkynyl groups. for example, nitrogen, Sulfur, or oxygen. Heterocyclic groups In certain embodiments, a straight-chain or branched may be saturated or unsaturated. Additionally, heterocyclic chain alkyl group may have 30 or fewer carbon atoms in its groups (such as pyrrolyl pyridyl, isoquinolyl, quinolyl, puri backbone, e.g., C-C for straight-chain or C-Clso for nyl, and furyl) may have aromatic character, in which case branched-chain. In certain embodiments, a straight-chain or 60 they may be referred to as "heteroaryl' or "heteroaromatic branched-chain alkyl group may have 20 or fewer carbon groups. atoms in its backbone, e.g., C-C for straight-chain or Unless otherwise stipulated, aryland heterocyclic (includ C-C for branched-chain, and more preferably 18 or fewer. ing heteroaryl) groups may also be substituted at one or more Likewise, preferred cycloalkyl groups have from 4-10 carbon constituent atoms. Examples of heteroaromatic and heteroali atoms in their ring structure, and more preferably have 4-7 65 cyclic groups may have 1 to 3 separate or fused rings with 3 to carbon atoms in the ring structure. The term “lower alkyl about 8 members per ring and one or more N, O, or Shet refers to alkyl groups having from 1 to 6 carbons in the chain, eroatoms. In general, the term "heteroatom' includes atoms US 8,642,801 B2 25 26 of any element other than carbon or hydrogen, preferred as phenoxathiinyl, carbazolyl, phenanthridinyl, acridinyl, examples of which include nitrogen, oxygen, Sulfur, and perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phosphorus. Heterocyclic groups may be saturated or unsat phenoxazinyl, and dibenzofuranyl groups. urated or aromatic. Some typical aryl groups include Substituted or unsubsti Examples of heterocycles include, but are not limited to, tuted 5- and 6-membered single-ring groups. In another acridinyl; azocinyl; benzimidazolyl; benzofuranyl; ben aspect, each Ar group may be selected from the group con Zothiofuranyl; benzothiophenyl; benzoxazolyl; benzthiaz sisting of substituted or unsubstituted phenyl, pyrrolyl, furyl, olyl; benztriazolyl; benztetrazolyl; benzisoxazolyl; ben thienyl, thiazolyl, isothiaozolyl, imidazolyl, triazolyl, tetra Zisothiazolyl; benzimidazolinyl; carbazolyl; 4aH-carbazolyl: Zolyl, pyrazolyl, oxazolyl, isooxazolyl pyridinyl, pyrazinyl, carbolinyl; chromanyl; chromenyl, cinnolinyl; decahydro 10 pyridazinyl, and pyrimidinyl groups. Further examples quinolinyl: 2H,6H-1.5.2-dithiazinyl; dihydrofuro2,3-bitet include substituted or unsubstituted phenyl, 1-naphthyl, rahydrofuran; furanyl; furazanyl; imidazolidinyl; imidazoli 2-naphthyl, biphenyl, 1-pyrrolyl 2-pyrrolyl, 3-pyrrolyl, nyl; imidazolyl: 1H-indazolyl; indolenyl; indolinyl: 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-ox indolizinyl; indolyl; 3H-indolyl; isobenzofuranyl: isochro azolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, manyl: isolindazolyl; isoindolinyl; isoindolyl; isoquinolinyl: 15 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, isothiazolyl; isoxazolyl; methylenedioxyphenyl; morpholi 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, nyl; naphthyridinyl; octahydroisoquinolinyl; oxadiazolyl; 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benz 1.2.3-oxadiazolyl: 1,2,4-oxadiazolyl: 1,2,5-oxadiazolyl: 1.3, imidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl 2-qui 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl; pyrim noxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl groups. idinyl; phenanthridinyl; phenanthrolinyl; phenazinyl; phe The term "amine' or "amino, as used herein, refers to an nothiazinyl; phenoxathiinyl; phenoxazinyl; phthalazinyl: unsubstituted or substituted moiety of the formula—NR'R''. piperazinyl; piperidinyl; piperidonyl: 4-piperidonyl; pipero in which R" and Rare each independently hydrogen, alkyl, nyl; pteridinyl; purinyl; pyranyl; pyrazinyl; pyrazolidinyl: aryl, or heterocyclyl, or R and R', taken together with the pyrazolinyl; pyrazolyl; pyridazinyl; pyridooxazole; pyri nitrogen atom to which they are attached, form a cyclic moi doimidazole; pyridothiazole; pyridinyl, pyridyl; pyrimidinyl: 25 ety having from 3 to 8 atoms in the ring. Thus, the term amino pyrrolidinyl; pyrrolinyl; 2H-pyrrolyl; pyrrolyl; quinazolinyl: includes cyclic amino moieties such as piperidinyl or pyrro quinolinyl; 4H-quinolizinyl; quinoxalinyl; quinuclidinyl; tet lidinyl groups, unless otherwise Stated. Thus, the term “alky rahydrofuranyl; tetrahydroisoquinolinyl; tetrahydroquinoli lamino” as used herein means an alkyl group having an amino nyl; tetrazolyl; 6H-1,2,5-thiadiazinyl: 1,2,3-thiadiazolyl: 1.2, group attached thereto. Suitable alkylamino groups include 4-thiadiazolyl: 1,2,5-thiadiazolyl: 1,3,4-thiadiazolyl: 30 groups having 1 to about 12 carbon atoms, preferably from 1 thianthrenyl; thiazolyl; thienyl; thienothiazolyl; thienoox to about 6 carbon atoms. The term amino includes com azolyl; thienoimidazolyl; thiophenyl; triazinyl: 1,2,3-triaz pounds or moieties in which a nitrogen atom is covalently olyl: 1,2,4-triazolyl: 1,2,5-triazolyl: 1,3,4-triazolyl; and Xan bonded to at least one carbon or heteroatom. The term thenyl. Preferred heterocycles include, but are not limited to, "dialkylamino' includes groups wherein the nitrogenatom is pyridinyl; furanyl; thienyl; pyrrolyl; pyrazolyl; pyrrolidinyl: 35 bound to at least two alkyl groups. The term “arylamino” and imidazolyl; indolyl; benzimidazolyl: 1H-indazolyl: oxazo "diarylamino' include groups wherein the nitrogen is bound lidinyl; benzotriazolyl; benzisoxazolyl: oxindolyl; benzox to at least one or two aryl groups, respectively. The term aZolinyl; and isatinoyl groups. Also included are fused ring “alkylarylamino” refers to an amino group which is bound to and spiro compounds containing, for example, the above at least one alkyl group and at least one aryl group. The term heterocycles. 40 "alkaminoalkyl refers to an alkyl, alkenyl, or alkynyl group A common hydrocarbon aryl group is a phenyl group hav substituted with an alkylamino group. The term "amide' or ing one ring. Two-ring hydrocarbonaryl groups include naph “aminocarbonyl includes compounds or moieties which thyl, indenyl, benzocyclooctenyl, benzocycloheptenyl, pen contain a nitrogen atom which is bound to the carbon of a talenyl, and aZulenyl groups, as well as the partially carbonyl or a thiocarbonyl group. hydrogenated analogs thereof Such as indanyl and tetrahy 45 The term “alkylthio’ refers to an alkyl group, having a dronaphthyl. Exemplary three-ring hydrocarbon aryl groups sulfhydryl group attached thereto. Suitable alkylthio groups include acephthylenyl, fluorenyl, phenalenyl, phenanthrenyl, include groups having 1 to about 12 carbon atoms, preferably and anthracenyl groups. from 1 to about 6 carbon atoms. Aryl groups also include heteromonocyclic aryl groups, The term “alkylcarboxyl as used herein means an alkyl i.e., single-ring heteroaryl groups, such as thienyl, furyl, pyra 50 group having a carboxyl group attached thereto. nyl, pyrrolyl, imidazolyl pyrazolyl pyridinyl, pyrazinyl, The term “alkoxy” as used herein means an alkyl group pyrimidinyl, and pyridazinyl groups; and oxidized analogs having an oxygen atom attached thereto. Representative thereof Such as pyridonyl, oxazolonyl, pyrazolonyl, isox alkoxy groups include groups having 1 to about 12 carbon aZolonyl, and thiazolonyl groups. The corresponding hydro atoms, preferably 1 to about 6 carbon atoms, e.g., methoxy, genated (i.e., non-aromatic) heteromonocyclic groups 55 ethoxy, propoxy, tert-butoxy and the like. Examples of alkoxy include pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazoli groups include methoxy, ethoxy, isopropyloxy, propoxy, nyl, pyrazolidinyl, pyrazolinyl, piperidyl and piperidino, pip butoxy, and pentoxy groups. The alkoxy groups can be Sub erazinyl, and morpholino and morpholinyl groups. stituted with groups such as alkenyl, alkynyl, halogen, Aryl groups also include fused two-ring heteroaryls such hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbo as indolyl, isoindolyl, indolizinyl, indazolyl, quinolinyl, iso 60 nyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, quinolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, cinnoli arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylami nyl, chromenyl, isochromenyl, benzothienyl, benzimida nocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, Zolyl, benzothiazolyl, purinyl, quinolizinyl, isoquinolonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino quinolonyl, naphthyridinyl, and pteridinyl groups, as well as (including alkyl amino, dialkylamino, arylamino, diary the partially hydrogenated analogs such as chromanyl, isoch 65 lamino, and alkylarylamino), acylamino (including alkylcar romanyl, indolinyl, isoindolinyl, and tetrahydroindolyl bonylamino, arylcarbonylamino, carbamoyl and ureido), groups. Aryl groups also include fused three-ring groups such imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sul US 8,642,801 B2 27 28 fates, alkylsulfinyl, Sulfonato, Sulfamoyl, Sulfonamido, nitro, and ureido), imino, Sulfhydryl, alkylthio, arylthio, thiocar trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an boxylate, Sulfates, alkylsulfinyl, Sulfonato, Sulfamoyl, Sul aromatic or heteroaromatic moieties. Examples of halogen fonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, Substituted alkoxy groups include, but are not limited to, alkylaryl, or an aromatic or heteroaromatic moiety. fluoromethoxy, difluoromethoxy, trifluoromethoxy, chlo Unless otherwise specified, the chemical moieties of the romethoxy, dichloromethoxy, trichloromethoxy, etc., as well compounds of the invention, including those groups dis as perhalogenated alkyloxy groups. cussed above, may be “substituted or unsubstituted.” In some The term “acylamino” includes moieties wherein an amino embodiments, the term “substituted” means that the moiety moiety is bonded to an acyl group. For example, the acy has substituents placed on the moiety other than hydrogen lamino group includes alkylcarbonylamino, arylcarbony 10 (i.e., in most cases, replacing a hydrogen), which allow the lamino, carbamoyl and ureido groups. molecule to perform its intended function. Examples of sub The terms “alkoxyalkyl”, “alkylaminoalkyl and “thio stituents include moieties selected from straight or branched alkoxyalkyl include alkyl groups, as described above, which alkyl (preferably C-C), cycloalkyl (preferably C-Cs), further include oxygen, nitrogen or Sulfur atoms replacing alkoxy (preferably C-C), thioalkyl (preferably C-C), alk one or more carbons of the hydrocarbon backbone. 15 enyl (preferably C-C), alkynyl (preferably C-C), hetero The term "carbonyl' or “carboxy” includes compounds cyclic, carbocyclic, aryl (e.g., phenyl), aryloxy (e.g., phe and moieties which contain a carbon connected with a double noxy), aralkyl (e.g., benzyl), aryloxyalkyl (e.g., bond to an oxygenatom. Examples of moieties which contain phenyloxyalkyl), arylacetamidoyl, alkylaryl, heteroaralkyl, a carbonyl include aldehydes, ketones, carboxylic acids, alkylcarbonyl and arylcarbonyl or other such acyl group, amides, esters, anhydrides, etc. heteroarylcarbonyl, and heteroaryl groups, as well as The term “ether' or “ethereal' includes compounds or (CRR")NR'R'" (e.g., -NH), (CRR"). CN (e.g., CN), moieties which contain an oxygen bonded to two carbon —NO, halogen (e.g., —F. —Cl. —Br, or—I), (CRR"). C atoms. For example, an ether or ethereal group includes (halogen) (e.g., —CF), (CRR")-CH(halogen), “alkoxyalkyl” which refers to an alkyl, alkenyl, or alkynyl (CRR"). CH(halogen), (CRR"). CONR'R", (CRR"). group Substituted with an alkoxy group. 25 (CNH)NR'R'', (CRR"). S(O)NR'R'', (CRR"). CHO, A “sulfonate” group is a —SOH or —SOX' group (CRR"). O(CRR")H, (CRR"). S(O) R' (e.g., bonded to a carbon atom, where X is a cationic counterion - SOH), (CRR"). O(CRR"). H (e.g., -CHOCH and group. Similarly, a “sulfonic acid compound has a —SOH –OCH), (CRR"). S(CRR"). H (e.g., -SH and or —SOX" group bonded to a carbon atom, where X is a —SCH), (CRR").OH (e.g., -OH), (CRR"). COR', cationic group. A "sulfate” as used herein is a —OSOH or 30 (CRR") (substituted or unsubstituted phenyl), (CRR"). —OSOX' group bonded to a carbon atom, and a “sulfuric (C-C cycloalkyl). (CRR"). COR' (e.g., —COH), and acid” compound has a —SOH or —OSOX" group bonded (CRR"). OR groups, wherein R and R" are each indepen to a carbon atom, where X" is a cationic group. According to dently hydrogen, a C-C alkyl, C-C alkenyl, C-C alky the invention, a Suitable cationic group may be a hydrogen nyl, oraryl group; or the side chain of any naturally occurring atom. In certain cases, the cationic group may actually be 35 amino acid. another group on the therapeutic compound that is positively In another embodiment, a substituent may be selected from charged at physiological pH, for example an amino group. straight or branched alkyl (preferably C-C), cycloalkyl A “counter ion' is required to maintain electroneutrality. (preferably C-Cs), alkoxy (preferably C-C), thioalkyl Examples of anionic counter ions include halide, triflate, (preferably C-C), alkenyl (preferably C-C), alkynyl (pref Sulfate, nitrate, hydroxide, carbonate, bicarbonate, acetate, 40 erably C-C), heterocyclic, carbocyclic, aryl (e.g., phenyl). phosphate, oxalate, cyanide, alkylcarboxylate, N-hydrox aryloxy (e.g., phenoxy), aralkyl (e.g., benzyl), aryloxyalkyl ySuccinimide, N-hydroxybenzotriazole, alkoxide, thioalkox (e.g., phenyloxyalkyl), arylacetamidoyl, alkylaryl, het ide, alkane Sulfonyloxy, halogenated alkane Sulfonyloxy, eroaralkyl, alkylcarbonyl and arylcarbonyl or other such acyl arylsulfonyloxy, bisulfate, oxalate, Valerate, oleate, palmi group, heteroarylcarbonyl, or heteroaryl group, (CRR"). tate, Stearate, laurate, borate, benzoate, lactate, citrate, male 45 oNR'R" (e.g., -NH), (CRR"). CN (e.g., -CN), NO, ate, fumarate. Succinate, tartrate, naphthylate mesylate, glu halogen (e.g., F. Cl, Br, or I), (CRR")o-oC(halogen) (e.g., coheptonate, or lactobionate. Compounds containing a —CF), (CRR"). CH(halogen), (CRR"). CH(halo cationic group covalently bonded to an anionic group may be gen), (CRR"). CONR'R'', (CRR") (CNH)NR'R", referred to as an “internal salt.” (CRR"). S(O)NR'R", (CRR"). CHO, (CRR").O The term "nitro” means —NO; the term “halogen' or 50 (CRR").H. (CRR"). S(O) R' (e.g., —SOH), “halogeno” or “halo' designates —F. —Cl, —Br or —I; the (CRR"). O(CRR").H (e.g., —CHOCH and term “thiol.” “thio.” or “mercapto' means SH; and the term —OCH), (CRR"). S(CRR"). H (e.g., —SH and “hydroxyl or “hydroxy' means –OH. —SCH), (CRR").H (e.g., -OH), (CRR"). COR', The term “acyl refers to a carbonyl group that is attached (CRR") (substituted or unsubstituted phenyl), through its carbon atom to a hydrogen (i.e., a formyl), an 55 (CRR") (C-C cycloalkyl), (CRR"). COR' (e.g., aliphatic group (e.g., acetyl), an aromatic group (e.g., ben —COH), or (CRR"). OR group, or the side chain of any Zoyl), and the like. The term “substituted acyl includes acyl naturally occurring amino acid; wherein R'and R" are each groups where one or more of the hydrogen atoms on one or independently hydrogen, a C-C alkyl, C-C alkenyl, C-Cs more carbon atoms are replaced by, for example, an alkyl alkynyl, or aryl group, or R' and R" taken together are a group, alkynyl group, halogen, hydroxyl, alkylcarbonyloxy, 60 benzylidene group or a —(CH2). O—(CH2) - group. arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, It will be understood that “substitution' or “substituted carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, with includes the implicit proviso that such substitution is in aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, accordance with the permitted valence of the substituted atom alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phos and the substituent, and that the substitution results in a stable phinato, cyano, amino (including alkylamino, dialkylamino, 65 compound, e.g., which does not spontaneously undergo arylamino, diarylamino, and alkylarylamino), acylamino (in transformation Such as by rearrangement, cyclization, elimi cluding alkylcarbonylamino, arylcarbonylamino, carbamoyl nation, etc. As used herein, the term "substituted” is meant to US 8,642,801 B2 29 30 include all permissible Substituents of organic compounds. In n-pentyl, n-heptyl, or n-octyl. In another embodiment, R' is a broad aspect, the permissible Substituents include acyclic t-butyl. In yet another alternate embodiment, R' is C7-Co and cyclic, branched and unbranched, carbocyclic and het bicycloalkyl or tricycloalkyl, such as, for example, tricyclo erocyclic, aromatic and nonaromatic Substituents of organic 3.3.1.0 decyl (or adamantyl), bicyclo[2.1.2]heptyl, or compounds. The permissible Substituents can be one or more. indolyl. In another alternate embodiment, R' is tetrahy In some embodiments, a “substituent may be selected dronaphthyl. from the group consisting of for example, halogeno, trifluo In one embodiment, L is —(CH) . In another further romethyl, nitro, cyano, C-C alkyl, C-C alkenyl, C-C, embodiment, Lis-(CH2) or -(CH2)5-. In yet another alkynyl, C-C alkylcarbonyloxy, arylcarbonyloxy, C-C, further embodiment, L is —(CH) . In yet another further alkoxycarbonyloxy, aryloxycarbonyloxy, C-C alkylcarbo 10 embodiment, L is substituted alkyl, e.g., —CH2— nyl, C-C alkoxycarbonyl, C-C alkoxy, C-C alkylthio. (CHOH)–CH-. arylthio, heterocyclyl, aralkyl, and aryl (including heteroaryl) In another embodiment, L' is CH-CH or absent. groups. Inafurther embodiment, R' is branched alkyl, e.g., t-butyl. In one embodiment, the invention pertains to compounds In another embodiment, R' is adamanyl. In another embodi 15 ment, R' is cyclic alkyl, e.g., cyclopropyl, cyclohexyl, cyclo of Formula I: heptyl, cyclo-octyl, etc. The cycloalkyl moieties may be Sub stituted further, e.g., with additional alkyl groups or other (I) groups which allow the molecule to perform its intended R2 function. In another embodiment, R' is alkyl substituted with a propargyl moiety (e.g., HC=C- ). In another embodiment, R NS-Y R" is cyclohexyl substituted with one or more methyl or n 11 L propargyl groups. In other embodiments, L' is a C-C alkyl linker group wherein: (e.g., —CH(CH)— or —(CH) . In a further embodi R" is a substituted or unsubstituted cycloalkyl, heterocy 25 ment, R' is phenyl. In certain embodiments, R' is substituted clic, aryl, arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic with a methoxy group. In other embodiments, L' is Cs, e.g., or tricyclic fused ring group, or a Substituted or unsubstituted -(CH2)4- or C(CH3)2 . In certain embodiments, L' is C2-Cio alkyl group; substituted, e.g., with an alkoxy, carboxylate ( COOH), R is selected from a group consisting of hydrogen, alkyl, benzyl, amido ( C=O NH ), or ester (C=O C. O) mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, 30 group. In certain embodiment, the ester group is a methyl, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, and ben ethyl, propyl, butyl, cyclohexyl, or benzyl ester. In other Zoimidazolyl; embodiments, the ester group may be propenyl. In other Y is SOX, OSO, X", or SSO, X"; embodiments, L' is substituted with a carboxylate group. In a X is hydrogen, a cationic group, or ester-forming group: further embodiment, R' is substituted with a substituted and 35 amido group, wherein the amido group is substituted with an each of L' and L is independently a substituted or unsub alkyl, e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl stituted C-C alkyl group or absent, or a pharmaceutically group. In another embodiment, the alkyl R' group is a sub acceptable salt thereof, provided that when R is alkyl, L' is stituted with a -C=O NH-OH, C=O NH, or an absent. amido group. In certain embodiments, the amido group is In a further embodiment, the invention pertains to com 40 Substituted with an alkyl (e.g., methyl, ethyl, propyl, butyl, pounds of Formula II: pentyl, hexyl, cyclohexyl, etc.), a benzyl or an aryl group. In another embodiment, the amido group is Substituted with a —CH(CH) group. R' itself may be substituted with a phe (II) R2 O nyl or may be branched or straight chain alkyl. In certain | || 45 embodiments, R' may also be substituted with a thioether N-(C)-(CH)-Y moiety. Examples of thioethers include S-Me, S-Et, etc. In / certain embodiments, the alkyl R' moiety is substituted with R-L both an aryl or a thioether moiety and an amido moiety. In other embodiments, the alkyl R' moiety may be substituted wherein: 50 with both a thioether and a carboxylate moiety. In other R" is a substituted or unsubstituted cyclic, bicyclic, tricy embodiments, alkyl R' groups are substituted with hydroxyl. clic, or benzoheterocyclic group or a substituted or unsubsti R" groups, e.g., alkyl R' groups, may also be substituted with tuted C-C alkyl group; both thioether and hydroxyl groups. In other embodiments, R is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, R" groups, e.g., alkyl R' groups are substituted with cyano cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, imidazolyl, 55 groups. Examples of R' groups including —CN moieties benzothiazolyl, benzoimidazolyl, or linked to R' to form a include —C(CH),CN, cyclohexyl substituted with one or heterocycle; more cyano groups, etc. Y is SOX, OSO, X", or SSO, X"; In other embodiments, alkyl R' groups are substituted with X" is hydrogen, a cationic group, or an ester forming moi aryl groups. The aryl groups may be substituted phenyl, for ety; 60 example. The substituted phenyl may be substituted with one m is 0 or 1; or more substituents such as hydroxy, cyano and alkoxy. In n is 1, 2, 3, or 4, other embodiments, alkyl R' groups are substituted with tet L is substituted or unsubstituted C-C alkyl group or razolyl or substituted or unsubstituted benzyl. In a further absent, or a pharmaceutically acceptable salt thereof, pro embodiment, L' is —C(CH3)2 -(CH2)—. In another vided that when R' is alkyl, L is absent. 65 embodiment, L' is —(C(CH4). CHOH-. In yet another Inafurther embodiment, R is hydrogen. In another further embodiment, L' is —(C(CH)-CH(OMe)-. In another embodiment, R' is straight chain alkyl, for example, ethyl, embodiment, R' is substituted or unsubstituted phenyl. In a US 8,642,801 B2 31 32 further embodiment, R' is para-substituted phenyl. Examples -continued of substitutents include but are not limited to fluorine, chlo rine, bromine, iodine, methyl, t-butyl, alkoxy, methoxy, etc. In other embodiment, R' is substituted at the meta position. N1\1Nso Examples of Substituents include methoxy, chloro, methyl, H t-butyl, fluoro, alkyl, alkoxy, iodo, trifluoroalkyl, methoxy, etc. In another embodiment, R' is phenyl substituted in the ortho position, with similar substituents. In another embodi ment, L' comprises a cycloalkyl moiety, e.g., cyclopentyl. In o 1N1\4 another embodiment, L' comprises an alkyenyl group and, 10 H MV optionally, a Substituted aryl group, with Substitutents similar O OH to those described about. In certain embodiments, R' is cyclopropyl or cyclohexyl. H In certain embodiments, the cyclopropyl or cyclohexyl group N-1N1\ is Substituted with an ether group or an alkyl group. In certain 15 > SOH further embodiments, the ether group is a benzyl ether group. H In another embodiment, wherein R' is alkyl, it is substi tuted with groups such as phenyl, or hydroxy. In other embodiments, the compound of the invention is > -- selected from the group consisting of: O > > N-1Nsa/ Yon N1 N1)-son 25 H r-so N-1N- SOH H H H N-- SOH 30 NS-1-N-SO3H

35 NS-1-N-SO3H

40 N-SONa N N-N-1-N-SO3H H

45

50

55

60

65 US 8,642,801 B2 33 -continued -continued O H HO Nu-1S-1 SOH

HCO HO N 1N1a SOH H

10

15

25

Br N-1N1 SOH

30 CHO

35

40

45

50

55

NaO Nu-1N1 SONa

60

Nu-1N1 SOH rx 65

US 8,642,801 B2 37 38 -continued -continued | H NN-1S-1 SOH afn,

10

NN-1N1 SOH 15

H

25

30

35

40

45

50

55

60

65

US 8,642,801 B2 41 42 -continued -continued

O

US 8,642,801 B2 43 44 -continued -continued

MeS

15

MeS

MeS

US 8,642,801 B2 47 48 -continued -continued O. O Me O O HO n-1N1\/ YoH

O \ ON 10 O O HO n-1N1\/ YoH OH Br O O 15

H V/ OH HO N-1N-1 S YoH

25 OH

30 OH

35

40

45

50

55

60

65 OMe

US 8,642,801 B2 55 56 -continued -continued O. O

YoH US 8,642,801 B2 57 58 -continued -continued

US 8,642,801 B2 59 60 -continued -continued

O. O O wn-1N1\/ YoH 1. N-1-1-4 / No O H

10 O. O M \, O O

H H 15 N-1-1-29 H M \, O O H

H

25

30 O |N-N-C O 35 I OH C O 1

40

45

OMe

O 50 "n-1-1-2 /Y,

H 55 X. O WN-NWYo "n-1-1-4 60 /\,

65

US 8,642,801 B2 65 66 -continued -continued

10

15

25

d) O O 4\, 30

>

45

50 O O

55 4x 2O N Y O 4. No N N-1N1so O H 60 O V y 1.N- 2 O H N s O. O 65 Y H US 8,642,801 B2 67 68 -continued double bond, provided that one of R. R. R. R. R. R. O R. R. R7 and R' is a moiety of the Formula IIIa: SFO (IIIa) 'C S ) / / RB (CH2) 10 RC RE RD

wherein: 15 m is 0, 1, 2, 3, or 4: R", R. R. R. and Rare independently selected from a group of hydrogen, halogen, hydroxyl, alkyl, alkoxyl, halo genated alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, cyano, thiazolyl, triazolyl, imidazolyl, tetrazolyl, ben Zothiazolyl, and benzoimidazolyl; and pharmaceutically acceptable salts and esters thereof, provided that said com pound is not 3-(4-phenyl-1,2,3,6-tetrahydro-1-pyridyl)-1- propanesulfonic acid. 25 In a further embodiment, n is 2, 3 or 4. In another embodiment, R'' is a salt-forming cation. Examples of salt forming cations include pharmaceutically acceptable salts described herein as well as lithium, Sodium, potassium, magnesium, calcium, barium, Zinc, iron, and 30 ammonium. In another embodiment, R' is an ester-forming group. An ester-forming group includes groups which when bound form an ester. Examples of Such groups include Sub stituted or unsubstituted alkyl, aryl, alkenyl, alkynyl, or cycloalkyl. In another embodiment, A is oxygen. 35 In another embodiment, Rand Rare taken together with and pharmaceutically acceptable salts, esters, and prodrugs the carbonatoms to which they are attached to form a double thereof. bond. In another embodiment, R. R. R. R. and R are In another embodiment, the invention pertains to com each hydrogen. R. R. R. and Rare each hydrogen and R. 40 is a halogen, such as fluorine, chlorine, iodine, or bromine pounds of Formula III: In another embodiment, R or R is a moiety of Formula IIIa. (III) In another embodiment, R. R. R. and Rare each hydro R4a R5 gen. In another further embodiment, R. R. R', and R' R R5a 45 O are each hydrogen. R3 In another, R is hydroxyl, cyano, acyl, or hydroxyl. In another further embodiment, R'' and A taken together R3a -cis---" are a natural or unnatural amino acid residue or a pharmaceu R7 R6 O tically acceptable salt or ester thereof. Examples of amino R7a R6a 50 acid residues include esters and salts of phenylalanine and leucine. wherein: In another embodiment, m is 0, 1, or 3. A is nitrogen or oxygen; Examples of compounds of Formula III include, but are not R'' is hydrogen, Salt-forming cation, ester forming group, limited to: -(CH2)-Q, or when A is nitrogen, A and R' taken together 55 may be the residue of a natural or unnatural amino acid or a salt or ester thereof; Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia c-( ) Or-ro Zolyl, or benzoimidazolyl: 60 X is 0, 1, 2, 3, or 4: n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; e R. R. R. R. R. R. R. R., R7 and R7 are each N-NN-1-N-SOs independently hydrogen, alkyl, mercaptoalkyl, alkenyl, alky NC nyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycar 65 bonyl, cyano, halogen, amino, tetrazolyl, or two R groups on ( ) \O-N- SOH adjacent ring atoms taken together with the ring atoms form a US 8,642,801 B2 69 70 -continued Zothiazolyl, and benzoimidazolyl, and pharmaceutically acceptable salts and esters thereof. In another embodiment, R'' is a salt-forming cation. Examples of salt forming cations include pharmaceutically acceptable salts described herein as well as lithium, Sodium, potassium, magnesium, calcium, barium, Zinc, iron, and ammonium. In another embodiment, R' is an ester-forming group. An ester-forming group includes groups which when N bound form an ester. Examples of Such groups include Sub 21 10 stituted or unsubstituted alkyl, aryl, alkenyl, alkynyl, or cycloalkyl. In another embodiment, A is oxygen. In another embodiment, m is 0 or 1. In another further N-1N1 SOH embodiment, n is 2, 3, or 4. In another further embodiment, OH R. R. Rand R7 are each hydrogen. R“, R, R, and R' Br N1\-1Nson 15 also may be hydrogen. Examples of R. R. R', R'', and R' include hydrogen. In another embodiment R. R. R'', R' HO are each hydrogen, and R' is a halogen, (e.g., fluorine, chlo C N1\1Nso rine, bromine, or iodine), nitro, or alkyl (e.g., methyl, ethyl, butyl). In another embodiment, A-R'' may be the residue of an amino acid, e.g., a phenylalanine residue. In another embodi ment, R. R', R'' and R'' are each hydrogen, and R is not hydrogen, e.g., halogen, e.g., fluorine, bromine, chlorine, or iodine. N1)-1Nso 25 In another embodiment, the compound is: and pharmaceutically acceptable salts, esters, and prodrugs 30 ()-\ / 1N1)-so 3 H thereof. In another embodiment, the invention pertains to com N 1N1 Sso H pounds of Formula IV: \ / 3

35 (IV) R9 R8 R4a R5 R R5a O R10 (CH-)-N 2)n N-(CH( 2)n --A-R' 40 R7 Y { R6a O R R12 R7a R6 wherein: 45 A is nitrogen or oxygen; and pharmaceutically acceptable salts, esters, and prodrugs R'' is hydrogen, salt-forming cation, ester forming group, thereof. -(CH2)-Q, or when A is nitrogen, A and R' taken together In another embodiment, the invention pertains to com may be the residue of a natural or unnatural amino acid or a 50 pounds of Formula V: salt or ester thereof; Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia Zolyl, or benzoimidazolyl: (V) X is 0, 1, 2, 3, or 4: R15 O n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 55 R. R. R. R. R. R. R7, and R7 are each indepen Re-s-cis--- dently hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, O cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbo nyl, cyano, halogen, amino, tetrazolyl, R' and R taken wherein: together, with the ring atoms they are attached to, form a 60 A is nitrogen or oxygen; double bond, or Rand R” taken together, with the ring atoms R'' is hydrogen, Salt-forming cation, ester forming group, they are attached to, form a double bond; -(CH2)-Q, or when A is nitrogen, A and R' taken together m is 0, 1, 2, 3, or 4: may be the residue of a natural or unnatural amino acid or a R. R. R', R', and R'' are independently selected from salt or ester thereof; a group of hydrogen, halogen, hydroxyl, alkyl, alkoxyl, halo 65 Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia genated alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, Zolyl, or benzoimidazolyl: aryl, cyano, thiazolyl, triazolyl, imidazolyl, tetrazolyl, ben X is 0, 1, 2, 3, or 4: US 8,642,801 B2 71 72 n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; acid (Idc), isonipecotic acid (Inp), meta-methyltyrosine aa is a natural or unnatural amino acid residue. (3-Me-Tyr), 1-naphthylalanine (1-Nal), 2-naphthylalanine m is 0, 1, 2, or 3: (2-Nal), para-nitrophenylalanine (4-NO-Phe), 3-nitroty R'' is hydrogen or protecting group; rosine (3-NO-Tyr), norleucine (Nle), norvaline (Nva), orni R" is hydrogen, alkyl or aryl, and pharmaceutically thine (Orn), ortho-phosphotyrosine (HPO-Tyr), octahy acceptable salts, esters and prodrugs thereof. droindole-2-carboxylic acid (Oic), penicillamine (Pen), In another embodiment, R'' is a salt-forming cation. pentafluorophenylalanine (Fs-Phe), phenylglycine (Phg), Examples of salt forming cations include pharmaceutically pipecolic acid (Pip), propargylglycine (Pra), pyroglutamic acceptable salts described herein as well as lithium, Sodium, acid (PGLU), sarcosine (Sar), tetrahydroisoquinoline-3-car potassium, magnesium, calcium, barium, Zinc, iron, and 10 ammonium. In another embodiment, R' is an ester-forming boxylic acid (Tic), thienylalanine, and thiazolidine-4-car group. An ester-forming group includes groups which when boxylic acid (thioproline. Th). Additionally, N-alkylated bound form an ester. Examples of Such groups include Sub amino acids may be used, as well as amino acids having stituted or unsubstituted alkyl, aryl, alkenyl, alkynyl, or amine-containing side chains (such as Lys and Orn) in which cycloalkyl. In another embodiment, A is oxygen. 15 the amine has been acylated or alkylated. In an embodiment, n is 2, 3 or 4. In certain embodiments, m Examples of compounds of the invention include, but are is 0. In certain embodiments, A-R'' is a residue of a natural not limited to: amino acid, or a salt or ester thereof. Examples of amino acid residues, include, but are not limited to, leucine or phenyla lanine residues, and pharmaceutically acceptable salts and esters thereof. Examples of possible esters include methyl, ethyl, and t-butyl. In another embodiment, m is 1. Examples of aa include H O O O natural and unnatural amino acid residues such as phenylala nine, glycine, and leucine. 25 HN Null-NY1N1 YoH In another embodiment, (aa) is a residue of phe-phe, oran ester thereof. | In certain embodiments, R' is hydrogen or substituted alkyl, e.g., arylalkyl. The term "unnatural amino acid refers to any derivative of a natural amino acid including D forms, 30 and C- and B-amino acid derivatives. It is noted that certain amino acids, e.g., hydroxyproline, that are classified as a non-natural amino acid herein, may be found in nature within a certain organism or a particular protein. Amino acids with H many different protecting groups appropriate for immediate 35 V/ use in the solid phase synthesis of peptides are commercially H-N N-1S-1 SS N ON-1 available. In addition to the twenty most common naturally occurringamino acids, the following examples of non-natural ls. H. O amino acids and amino acid derivatives may be used accord ing to the invention (common abbreviations in parentheses): B-alanine (B-ALA), Y-aminobutyric acid (GABA), 2-ami nobutyric acid (2-Abu), C.B-dehydro-2-aminobutyric acid (8-AU), 1-aminocyclopropane-1-carboxylic acid (ACPC), aminoisobutyric acid (Aib), 2-amino-thiazoline-4-carboxy H O. O lic acid, 5-aminovaleric acid (5-Ava), 6-aminohexanoic acid 45 \/ O (6-AhX), 8-aminooctanoic acid (8-Aoc), 11-aminounde HN' n-1N1 n-1 canoic acid (11-Aun), 12-aminododecanoic acid (12-Ado), 2-aminobenzoic acid (2-Abz), 3-aminobenzoic acid (3-Abz), C O H O 4-aminobenzoic acid (4-AbZ), 4-amino-3-hydroxy-6-methyl heptanoic acid (Statine, Sta), aminooxyacetic acid (Aoa), 50 2-aminotetraline-2-carboxylic acid (ATC), 4-amino-5-cyclo hexyl-3-hydroxypentanoic acid (ACHPA), para-aminophe H O O nylalanine (4-NH-Phe), biphenylalanine (Bip), para-bro l \/ ONa mophenylalanine (4-Br-Phe), ortho-chlorophenylalanine (2- H-N nu1N1 NN Cl-Phe), meta-chlorophenylalanine (3-Cl-Phe), para 55 chlorophenylalanine (4-Cl-Phe), meta-chlorotyrosine (3-Cl Tyr), para-benzoylphenylalanine (Bpa), tert-butylglycine (TLG), cyclohexylalanine (Cha), cyclohexylglycine (Chg), 2,3-diaminopropionic acid (Dpr), 2,4-diaminobutyric acid (Dbu), 3,4-dichlorophenylalanine (3,4-Cl-Phe), 3,4-difluo 60 rorphenylalanine (3.4-F-Phe), 3.5-diiodotyrosine (3.5-I- Tyr), ortho-fluorophenylalanine (2-F-Phe), meta-fluorophe H O. O nylalanine (3-F-Phe), para-fluorophenylalanine (4-F-Phe), \/ O meta-fluorotyrosine (3-F-Tyr), homoserine (Hse), homophe HN' nu1N1 N nylalanine (Hfe), homotyrosine (Htyr), 5-hydroxytryptophan 65 (5-OH-Trp), hydroxyproline (Hyp), para-iodophenylalanine C O H O (4-I-Phe), 3-iodotyrosine (3-I-Tyr), indoline-2-carboxylic US 8,642,801 B2 73 74 -continued In another embodiment, R'' is a salt-forming cation. Examples of salt forming cations include pharmaceutically acceptable salts described herein as well as lithium, Sodium, potassium, magnesium, calcium, barium, Zinc, iron, and 1n 1a 1 -CO, Na 5 ammonium. In a further embodiment, the salt is a sodium salt. e N SO-N Y3. In a further, embodiment, A is oxygen. In another embodiment, Y is oxygen or sulfur, and R is absent. 10 In another embodiment, Y is oxygen and R is absent. Examples of R' include benzyl, aryl (e.g., phenyl), alkyl, cycloalkyl (e.g., adamantyl), etc. In other embodiment, Y is nitrogen and R is hydrogen. In other embodiment, R is benzyl. In another further embodiment, R” and R are 15 linked to form a pyridyl ring. In another embodiment, Y is O. O sulfur. HN2n-1N1 \/ O N-1 Examplesp les of compoundsp ds of the iinvention, ion, includinclude

H O 2O and pharmaceutically acceptable salts, esters, and prodrugs O ros. thereof. H OH In another embodiment, the invention pertains, at least in 25 part, to compounds of Formula VI: O N 1n 1\so (VI) H R22 30 O R21V M O 1n-1\-1 SONa Y2-C-N-(CH)-S-A-RI / H R20 R23 R19 O 35 O wherein: n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; H A is oxygen or nitrogen; R'' is hydrogen, Salt-forming cation, ester forming group, 40 -(CH2)-Q, or when A is nitrogen, A and R' taken together NH NH SO Na may be the residue of a natural or unnatural amino acid or a N-1N1''' salt or ester thereof; Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia Zolyl, or benzoimidazolyl: 45 X is 0, 1, 2, 3, or 4: R" is hydrogen, alkyl or aryl; O Y' is oxygen, sulfur, or nitrogen; Y is carbon, nitrogen, or oxygen; R" is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, 50 SONa alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetra Zolyl, imidazolyl, benzothiazolyl, or benzoimidazolyl: R’ is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imi- N SO3Na dazolyl, benzothiazolyl, benzoimidazolyl, or absent if Y is 55 n1N1''' OXygen; R’ is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, O cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imi- O dazolyl, benzothiazolyl, benzoimidazolyl: or R’ is hydro- ls gen, hydroxyl, alkoxy or aryloxy if Y is nitrogen; or R’ is 60 1N1\-1N 1N1)-so Na absent if Y is oxygen or sulfur, or RandR may be linked 3 to form a cyclic moiety if Y is nitrogen; H H R’ is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, l alkynyl,kynyl, cycloalkyl, aryl, arylalkyl,arylalky thiazolyl,y triazolyl,y tetra N1 N1-1NsoN Zolyl, imidazolyl, benzothiazolyl, or benzoimidazolyl, or 65 absent if Y is nitrogen or oxygen; H H or pharmaceutically acceptable salts thereof. US 8,642,801 B2 75 76 -continued embodiments, two or more R substituents can be linked to form a fused ring (e.g., to form a methylendioxyphenyl moi ety). Examples of compounds of the invention include:

No 1n 1a NHS-1a-SONa 10 NH SOH

O O 1. O 15 NH NH1N1 Nson, N1N1 Nson > l ON NH N1,N1H Nson \

HO 25 r-son H and pharmaceutically acceptable salts, esters, and prodrugs HO thereof. In another embodiment, the invention pertains to com O pounds of Formula VII: 30 1. N11 N1)-son (VII) y ON / \ (CH)-N-(CH2)-S-A-RI 35 (R-G1 (— e | r-son O H 40 / O wherein: n is 2, 3, or 4: A is oxygen or nitrogen; R'' is hydrogen, Salt-forming cation, ester forming group, 45 -(CH2)-Q, or when A is nitrogen, A and R' taken together and pharmaceutically acceptable salts, esters, and prodrugs may be the residue of a natural or unnatural amino acid or a thereof. salt or ester thereof; Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothia Other compounds of the invention include Zolyl, or benzoimidazolyl: 50 X is 0, 1, 2, 3, or 4: G is a direct bond or oxygen, nitrogen, or Sulfur, Z is 0, 1, 2, 3, 4, or 5: m is 0 or 1; R" is selected from a group consisting of hydrogen, alkyl, 55 mercaptoalkyl, alkenyl, alkynyl, aroyl, alkylcarbonyl, ami noalkylcarbonyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triaz olyl, imidazolyl, benzothiazolyl, and benzoimidazolyl: each R is independently selected from hydrogen, halo gen, cyano, hydroxyl, alkoxy, thiol, amino, nitro, alkyl, aryl, 60 carbocyclic, or heterocyclic, and pharmaceutically accept able salts, esters, and prodrugs thereof. In one embodiment, R'' is hydrogen. In another, A is oxy gen. For example, n may be 3 and m may be 1. In other embodiments, R is hydrogen or benzyl. 65 In certain embodiments, Z is 0, 2, or 3. In others, R is hydroxyl or alkoxy, e.g., methoxy, ethoxy, etc. In certain US 8,642,801 B2 77 78 -continued -continued

10 OH O and pharmaceutically acceptable salts, esters, and prodrugs thereof. NH2 15 The invention pertains to both salt forms and acid/base NH forms of the compounds of the invention. For example, the invention pertains not only to the particular salt forms of I N SOH compounds shown herein as salts, but also the invention NH includes other pharmaceutically acceptable salts, and the acid and/or base form of the compound. The invention also per HN ls S 1N1\ SN/ tains to salt forms of compounds shown herein. / OH Compounds of the invention are also shown in Table 2 below. TABLE 2

ID Structure? Name of Compound

NH1N1 Nso S 2-phenyl-1-sulfopropyl-1,2,3,6-tetrahydropyridine

21 N -- 4-(3-phenylpropyl)-1-sulfopropylpyridine

21 Nu-1N1 SOH 4-(3-phenylpropyl)-1-sulfopropyl-2,3,6-tetrahydropyridine

NC N-1 n-1 SOH 3-(4-cyano-4-phenylpiperidin-1-yl)-1-propanesulfonic acid -()-Or-ro 3-4-(4-fluorophenyl)-1,2,3,6-tetrahydropyridin-1-yl)-1-propanesulfonic acid

OH B -() X)^-so 3-4-(4-bromophenyl)-4-hydroxypiperidin-1-yl)-1-propanesulfonic acid

US 8,642,801 B2 83 84 TABLE 2-continued

ID Structure/Name of Compound AC -Irrison.O H OH

N-benzyloxycarbonyl-3-amino-2-hydroxy-1-propanesulfonic acid sodium salt

AD O p-- 1N1-S-SO Na H

N-benzyloxycarbonyl-4-amino-1-butanesulfonic acid sodium salt AE -Ir-nos,O H

N-benzyloxycarbonyl-3-amino-1-propanesulfonic acid sodium salt

AF n-1N1 SOH

O

3-(benzhydrylamino)carbonylamino-1-propanesulfonic acid

AG H N-1S-1 SONa

O 3-(phenylacetyl)amino-1-propanesulfonic acid, sodium salt

AH O

H sn-nos,H 3-(benzylamino)carbonylamino-1-propanesulfonic acid, sodium salt AI ~~~~O 3- {(hexylamino)carbonylamino-1-propanesulfonic acid, sodium salt AJ is-n-ros.O

3-(dodecylamino)carbonylamino-1-propanesulfonic acid, sodium salt US 8,642,801 B2 85 86 TABLE 2-continued

ID Structure/Name of Compound AK A. 2-.O H H 3- (adamantylamino)carbonylamino-1-propanesulfonic acid, sodium salt

AL NH N-1N1 SONa

O 3-2-(4-isobutylphenyl)propanoyl)amino-1-propanesulfonic acid, sodium salt

AM S NH1N11 N1)-soN

3-(benzylamino)carbonothioylamino-1-propanesulfonic acid, sodium salt

AU

H 1 -CO, Na N11 Nso N w

N-(3-dibenzylamino-1-propanesulfonyl)-L-phenylalanine, sodium salt

AV

NN-1N1 SOH 3-dibenzylamino-1-propanesulfonic acid AW CO?O NH1N1 Nso 3-((1,3-benzodioxol-5-yl)methyl)amino]-1-propanesulfonic acid

1. O NH 1N1\ SOH No 3-(3,4-dimethoxybenzylamino)-1-propanesulfonic acid AY / NH1N1 Nso

O / ON 3-(3,4,5-trimethoxybenzylamino)-1-propanesulfonic acid US 8,642,801 B2 87 88 TABLE 2-continued

ID Structure/Name of Compound NH1N1 Nson

O O s

3-(2,3-dimethoxybenzylamino)-1-propane Sulfonic acid BA NO NH1N1 Nso

3-(3,5-dimethoxybenzylamino)-1-propane Sulfonic acid

BB O h / CroO 3-(2,4-dimethoxybenzylamino)-1-propanesulfonic acid

BC HO 1N1S SOH H HO 3-(3,4-dihydroxybenzylamino)-1-propanesulfonic acid

BW

O H r-son 3-(1-adamantyl)amino-1-propanesulfonic acid

H YN 1n 1a SOH

3-(t-butyl)amino-1-propanesulfonic acid

BY H -- SOH

3-(2-norbornyl)amino-1-propanesulfonic acid

H N-- SOH

3-(2-adamantyl)amino-1-propanesulfonic acid CC N-1a1n 1 SOH 5-amino-1-pentanesulfonic acid US 8,642,801 B2 89 90 TABLE 2-continued

ID Structure/Name of Compound CD 2NS-11N1 SOH 6-amino-1-hexanesulfonic acid

CE H N-1N1 SOH 3-isobutylamino-1-propanesulfonic acid

CG H N N-1N1 SOH

3-isopropylamino-1-propanesulfonic acid

CH H N- N-1N1 SOH 3-isoamylamino-1-propanesulfonic acid

CI D- NS-1-N-SO3H 3-(cyclopropylamino)-1-propanesulfonic acid

H N-1N-1 SOH 3-(cyclopentylamino)-1-propanesulfonic acid

CK H -- SOH

3-(cycloheptylamino)-1-propanesulfonic acid

CL

H O N-(3-aminopropane-1-sulfonyl)-phenylalanine, ethyl ester

CM H N YSO.Na

cyclopentylsulfamic acid, Sodium salt

CN N-SONa H

cycloheptylsulfonic acid, Sodium salt US 8,642,801 B2 91 92 TABLE 2-continued

ID Structure/Name of Compound

CO O

I N SOH 4-iodo-N-(3-sulfopropyl)-L-phenylalanine amide CV 1N1 S-1Nso 3-(ethylamino)-1-propanesulfonic acid

CY

HC

3-(3,5-dimethyl-1-adamantylamino)-1-propanesulfonic acid

DC OH NH-N- SOH

3-cyclohexylamino-2-hydroxy-1-propanesulfonic acid

DD H NS-1-N-SO3H

3-(3-pentyl)amino-1-propanesulfonic acid

DE

OH

DG H -x- SOH 3-(tert-amyl)amino-1-propanesulfonic acid

DH H rx NS-1-N-SO3H 3-(1,1-dimethyl-2-hydroxyethyl)amino-1-propanesulfonic acid

US 8,642,801 B2 99 100 TABLE 2-continued

ID Structure/Name of Compound

EI H ar N-1N1 SOH 3-(2-methoxy-1-methylethyl)amino-1-propanesulfonic acid

H N-- SOH

CH2OH 3-(1R)-2-benzyl-1-hydroxyethylamino-1-propanesulfonic acid EK N-olH

3-(1S)-2-benzyl-1-hydroxyethylamino-1-propanesulfonic acid

EL

EN

3-(N-methyl-N-tert-butylamino)-1-propanesulfonic acid

EO IIIOH

Nin 1 N1 SOH 3-(1R,2S)-2-hydroxyindan-1-amino-1-propanesulfonic acid

EP ----H SOH CH2OH 3-(1S)-1-(hydroxymethyl)-2-methylpropylamino-1-propanesulfonic acid -----H 4 3-(1S)-1-carbamoyl-2-methylpropylamino-1-propanesulfonic acid

ER H X N-1N1 SOH 4-(tert-butylamino)-1-butanesulfonic acid US 8,642,801 B2 101 102 TABLE 2-continued

ID Structure/Name of Compound ES > -->H SOH 4-(tert-butylamino)-2-butanesulfonic acid

ET

N1H N1)-son

3-(2,2-diphenylethyl)amino-1-propanesulfonic acid

EV

r O O N11SNOH 3-(4-mexiletino)-1-propanesulfonic acid

EW N O O O

3-(1-benzyl-2-methoxyethyl)amino-1-propanesulfonic acid

EY

H N-1S-1 SOH

EZ

H N-1N1 SOH

FA H H3CO N n-1N1 SOH

N H

FH N 1N1S SONa

FL

N1N1)-sonH US 8,642,801 B2 103 104 TABLE 2-continued

ID Structure/Name of Compound FM

H or.O NH2 CH3

O' N1,N1H Nson CHO

FO

OH no1 N s1 H M \, O O O

FP

FR

FS

FT

FU US 8,642,801 B2 105 106 TABLE 2-continued

ID Structure/Name of Compound

H HC* N1 N N-1N1'''SOH

4. NH2

CH3 croH

OH FY

GA

GB

GC US 8,642,801 B2 107 108 TABLE 2-continued

ID Structure/Name of Compound GD OCH2Ph

'N1\1\sonH

GE H NS-1a-SO3H

GF

GH

GI

G

GK

GL

GM US 8,642,801 B2 109 110 TABLE 2-continued

ID Structure/Name of Compound GN

GO

GP

GR

GS

GT

GU

GZ. US 8,642,801 B2 111 112 TABLE 2-continued

ID Structure/Name of Compound

HA O

N-1N1 gaOH o21 No

HB

HC

HD

HE

HF

HG

HI

H

HO Nu-1N1 la OH US 8,642,801 B2 113 114 TABLE 2-continued

ID Structure/Name of Compound

HK H O leo n

1. rO

HL

HM

HN

HO

HP

HR US 8,642,801 B2 115 116 TABLE 2-continued

ID Structure/Name of Compound

HS O / =o

HT

HU

HV

HY

HZ US 8,642,801 B2 117 118 TABLE 2-continued

ID Structure/Name of Compound

IA O O --~~4/ So -- H O

IB O

OESEO

IC

O O O 41, N 1-14M. Yo H O H O

ID

O

N O NH N y-s-s H1 nu-1N1 \,

IE O V2O 4. US 8,642,801 B2 119 120 TABLE 2-continued

ID Structure/Name of Compound IF N/ O\eo / /N n-1N1 SY, o= | O

IG

O Nt / so

IH Nt

O N-n-,\eo II ulO O e N D

O-SEO | O Os-O 21 N V 2O2 N N-n-

IK US 8,642,801 B2 121 122 TABLE 2-continued

ID Structure/Name of Compound

IL V 29 Nt N-n-,

IM V 2O Nt N-n-,

IN V 29 Br NN-1N1 \,

IO

O )- IP

IR

IS

IT US 8,642,801 B2 123 124 TABLE 2-continued

ID Structure/Name of Compound IU s/

IV

IW

IX

IY

IZ.

JA

JB US 8,642,801 B2 125 126 TABLE 2-continued

ID Structure/Name of Compound

JD

JE

JG

JH

JI

JK US 8,642,801 B2 127 128 TABLE 2-continued

ID Structure/Name of Compound JL HO -l CH

H ~so

JM

N- SONa

JO N SONa 1. ~ 3

JP

H N- SONa

H N-- SOH o-s, JS ~.

S-N-nos,/ OH

1 H r^o O H

H >r.OH US 8,642,801 B2 129 130 TABLE 2-continued

ID Structure/Name of Compound

OH H N SONa

H r^so OH -Nnson OH

KB

H rrno. OH

KH H N-- SOH CH3

KI OH N--H SOH

K OH N--H SOH

KK OH H --- SOH

KL OH H N-'-- SOH NoH

KM O N-> CH3 SOH

US 8,642,801 B2 133 134 TABLE 2-continued

ID Structure/Name of Compound

O OS) / /

KY

N H LA V -O 2.

O

O N \ N LC V -O 4.

O

O \ N N

LD H >{ N-1N1a SOH

LE 1n N 11-N-SO3H <-{N-1S-1Nso

N-1N1a / /S No US 8,642,801 B2 13 5 136 TABLE 2-continued

ID Structure/Name of Compound LI H NN-1N1a SOH

L 1N-1-1-1-1-1--"

LK

LL

LM

LN

LO

LP

NE

NG

OMe US 8,642,801 B2 137 138 TABLE 2-continued

ID Structure/Name of Compound

NH

OH N 1N1-( H M No O

NI MeO

OH N 1--1a( H /A So

NJ OH O | N-1-is OH

NK Me

OH N 1-1Ns H /A So

NL F

NH 1N1)-so OH

It should be noted that in the above table and throughout the methods of using the compounds described in WO 00/64420 application when an atom is shown without hydrogens, but and WO 96/28187 for methods described in this application, hydrogens are required or chemically necessary to form a which are not described in WO 00/64420 and WO 96/28187. stable compound, hydrogens should be inferred to be part of 60 Both of WO 00/64420 and WO 96/28.187 are incorporated by the compound. In one embodiment, the invention does not pertain to the reference herein in their entirety. compounds described in WO 00/64420 and WO 96/28187. In In another embodiment, the invention pertains to methods this embodiment, the invention does not pertain to methods of of the invention which use and pharmaceutical compositions using the compounds described in WO 00/64420 and WO 65 comprising, the compounds of Table 2A. In another embodi 96/28187 for the treatment of diseases or disorders described ment, the compounds of the invention do not include the therein. In a further embodiment, the invention pertains to compounds of Table 2A. US 8,642,801 B2 139 140 TABLE 2A

MeO

MeO N-1N1 SOH

BzNHCH2CHCHSONa

N Or-so HO

OH OH

O

N 11a-SOsk

HN SOH COOH

N-1a-1 SONa

H Bn -O-clickson

2 N HC SOH CH3 O CH3 US 8,642,801 B2 141 142 TABLE 2A-continued

O

CrO Ol N CHO Clu- SOH

AcNHCHCH-CHSONa

O

-19HC H US 8,642,801 B2 143 144 TABLE 2A-continued

N

CH HC1 NN-1N1 SONa

HO r-son H

H Ph1 NN-1N1 SONa

CC SONa

ON-1N1n 1\ 1N1a SOH H r-so H ~~~~~ H

On 11a 1N1a SOH H

SONa OCH Ns O N SONa OH NH NH 2 OH NaOS Ns N

SONa US 8,642,801 B2 145 146 TABLE 2A-continued

N N H3C S S CH3 SONa N=N. SO3Na N H -- N

2 HO n-r-so

HO s 1n 1 N SOH

HO H

HO , 1N1a N SOH

US 8,642,801 B2 149 150 TABLE 2A-continued US 8,642,801 B2 151 152 TABLE 2A-continued

OH N Nukot

HO OH US 8,642,801 B2 153 154 TABLE 2A-continued H - O ( ) / -I-1N1 son, US 8,642,801 B2 155 156 TABLE 2A-continued

OH N-- SOH

O

N1N1H Nson

NaOS OMe O X

NHCH2CH2CH2SOH

NaOSOCH2(CH2)CH2OSONa

HOCHCH- N-CHCHSOH

HOCHCH- N-CHCHSONa

N-CHCHCHSOH

O N-CHCHCHSONa

HOCH2CH2CHCHSONa

NaOS SONa

S 4./V \,

SOK HO

OH SOK US 8,642,801 B2 158 TABLE 2A-continued SONa

HO SONa OH

CHSONa O

CHSONa NaOSCH2CHCH CHSONa OSONa OSONa OSONa CHOH HO O NaOSO NaOSO OMe

P1Y-NuoNaOSO NaOSO OMe NaOS O HO OMe

OH on--is OSONa NaOS O OH 7,O CHC(CH2OSONa) NH2CH2CH2CH2SONa HC(CHOSONa) NHC(CH2OSONa) NH2CH2CH2OSOH NaOSNHCHCHOSONa O

NaOS HN

NH SONa US 8,642,801 B2 159 160 TABLE 2A-continued

CH2OH OH

O CH O OR RO O RO CH2OH OR OR

OSONa PhCH2O OSONa

NaOSNHCHCHCHOSONa HN(CH2CH2OSONa) H2NCHCH-CHOSO-Na. NaOSN(CH2CH2OSONa) H2NCHCHSOH NaOSOCHCHCHSONa

SONa

SONa

SONa MeO

OMe SONa

SONa

sMeO SONa

SOK MeO

OMe SOK

NaOSO BO NaOSO OSONa OB OSONa US 8,642,801 B2 161 162 TABLE 2A-continued O OSONa

O OSONa

OH OSONa OH

OorSOH NH2 s NH2

SOH OSONa

OSONa on----- NaOSO OSONa CHCH2CH2CH2SONa CH(CH2)CHSONa CH-CH-CH SONa CH-CH-CH-CH-CH SONa ---->OR OR OR R = SONa n--- SONa US 8,642,801 B2 163 164 TABLE 2A-continued

SONa

US 8,642,801 B2 167 168 TABLE 2A-continued

NaOS I COOH O CH-CH V NH2 NaOS I or-rr-roos, OSONa

OH NaOS N-N- SONa

NaOC H

OSONa

O

CH-N --> SONa H Soo-K)-os. OSONa

NaOSO OSONa

I O COOH M CH-CH NaOS O M NH2 NaO. S I

SONa H CH-N N-CO SONa

OH NaOS N-N- SONa

SONa SONa SONa

SONa US 8,642,801 B2 169 170 TABLE 2A-continued NaOS n-n-n- SONa SONa ic--ison,CHSONa CHSONa NaOS n-n-n-n- SONa NaOS SONa

OH "N-N- SOH

NH SONa

2 CN NHCH2SONa

2 N NaOC ros. in--- SONa os---to SONa

NaOC SONa

SONa NaOC CONa No.1\-1 SO3Na Naoc1N1 NsoN

O

NaOS O

NaOS US 8,642,801 B2 171 172 TABLE 2A-continued NaOS o-( )-( ) NaOS CHCH2CH2SONa CHCH2SONa CHCH2CH2CH2CH2SONa

It should be understood that the use of any of the com 15 therapy. The Subject may also be a male human. In another pounds described herein or in the applications identified in embodiment, the subject is under 40 years old. “The Related Applications' Section is within the scope of the A Subject may be a human at risk for Alzheimer's disease, present invention and is intended to be encompassed by the e.g., being over the age of 40 or having a predisposition for present invention and each of the applications are expressly Alzheimer's disease Alzheimer's disease predisposing fac incorporated herein at least for these purposes, and are fur tors identified or proposed in the scientific literature include, thermore expressly incorporated for all other purposes. among others, a genotype predisposing a Subject to Alzhe Subjects and Patient Populations imer's disease; environmental factors predisposing a subject The term “subject includes living organisms in which to Alzheimer's disease; past history of infection by viral and amyloidosis can occur, or which are susceptible to amyloid bacterial agents predisposing a Subject to Alzheimer's dis diseases, e.g., Alzheimer's disease, Down's syndrome, CAA, 25 ease; and vascular factors predisposing a Subject to Alzhe dialysis-related (BM) amyloidosis, secondary (AA) amyloi imer's disease. A Subject may also have one or more risk factors for cardiovascular disease (e.g., atherosclerosis of the dosis, primary (AL) amyloidosis, hereditary amyloidosis, coronary arteries, angina pectoris, and myocardial infarction) diabetes, etc. Examples of Subjects include humans, chick or cerebrovascular disease (e.g., atherosclerosis of the intrac ens, ducks, peking ducks, geese, monkeys, deer, cows, rab 30 ranial or extracranial arteries, stroke, Syncope, and transient bits, sheep, goats, dogs, cats, mice, rats, and transgenic spe ischemic attacks), Such as hypercholesterolemia, hyperten cies thereof. Administration of the compositions of the sion, diabetes, cigarettesmoking, familial or previous history present invention to a subject to be treated can be carried out of coronary artery disease, cerebrovascular disease, and car using known procedures, at dosages and for periods of time diovascular disease. Hypercholesterolemia typically is effective to modulate amyloid aggregation or amyloid-in 35 defined as a serum total cholesterol concentration of greater duced toxicity in the subject as further described herein. An than about 5.2 mmol/L (about 200 mg/dL). effective amount of the therapeutic compound necessary to Several genotypes are believed to predispose a subject to achieve a therapeutic effect may vary according to factors Alzheimer's disease. These include the genotypes such as Such as the amount of amyloid already deposited at the clini presenilin-1, presenilin-2, and amyloid precursor protein cal site in the Subject, the age, sex, and weight of the Subject, 40 (APP) missense mutations associated with familial Alzhe and the ability of the therapeutic compound to modulate imer's disease, and O-2-macroglobulin and LRP-1 geno amyloid aggregation in the Subject. Dosage regimens can be types, which are thought to increase the risk of acquiring adjusted to provide the optimum therapeutic response. For sporadic (late-onset) Alzheimer's disease. E. van Uden, et al., example, several divided doses may be administered daily or J. Neurosci. 22(21),9298-304 (2002); J. J. Goto, et al., J. Mol. the dose may be proportionally reduced as indicated by the 45 Neurosci. 19(1-2), 37-41 (2002). Another genetic risk factor exigencies of the therapeutic situation. for the development of Alzheimer's disease are variants of In certain embodiments of the invention, the subject is in ApoE, the gene that encodes apolipoprotein E (particularly need of treatment by the methods of the invention, and is the apoE4 genotype), a constituent of the low-density lipo selected for treatment based on this need. A subject in need of protein particle. WJ Strittmatter, et al., Annu. Rev. Neurosci. treatment is art-recognized, and includes subjects that have 50 19, 53-77 (1996). The molecular mechanisms by which the been identified as having a disease or disorder related to various ApoE alleles alter the likelihood of developing Alzhe amyloid-deposition or amyloidosis, has a symptom of such a imer's disease are unknown, however the role of ApoE in disease or disorder, or is at risk of such a disease or disorder, cholesterol metabolism is consistent with the growing body and would be expected, based on diagnosis, e.g., medical of evidence linking cholesterol metabolism to Alzheimer's diagnosis, to benefit from treatment (e.g., curing, healing, 55 disease. For example, chronic use of cholesterol-lowering preventing, alleviating, relieving, altering, remedying, ame drugs such as statins has recently been associated with a lower liorating, improving, or affecting the disease or disorder, the incidence of Alzheimer's disease, and cholesterol-lowering symptom of the disease or disorder, or the risk of the disease drugs have been shown to reduce pathology in APP trans or disorder). genic mice. These and other studies Suggest that cholesterol In an exemplary aspect of the invention, the Subject is a 60 may affect APP processing. ApoE4 has been suggested to human. For example, the subject may be a human over 30 alter AB trafficking (in and out of the brain), and favor reten years old, human over 40 years old, a human over 50 years tion of AB in the brain. ApoE4 has also been Suggested to old, a human over 60 years old, a human over 70 years old, a favor APP processing toward A? formation. Environmental human over 80 years old, a human over 85 years old, a human factors have been proposed as predisposing a Subject to over 90 years old, or a human over 95 years old. The subject 65 Alzheimer's disease, including exposure to aluminum, may be a female human, including a postmenopausal female although the epidemiological evidence is ambiguous. In addi human, who may be on hormone (estrogen) replacement tion, prior infection by certain viral or bacterial agents may US 8,642,801 B2 173 174 predispose a subject to Alzheimer's disease, including the In determining the CDR, a Subject is typically assessed and herpes simplex virus and chlamydia pneumoniae. Finally, rated in each of six cognitive and behavioural categories: other predisposing factors for Alzheimer's disease can memory, orientation, judgement and problem solving, com include risk factors for cardiovascular or cerebrovascular dis munity affairs, home and hobbies, and personal care. The ease, including cigarette Smoking, hypertension and diabetes. assessment may include historical information provided by At risk for Alzheimer's disease' also encompasses any other the subject, or preferably, a corroborator who knows the sub predisposing factors not listed above or as yet identified and ject well. The subject is assessed and rated in each of these includes an increased risk for Alzheimer's disease caused by areas and the overall rating, (0, 0.5, 1.0, 2.0 or 3.0) deter head injury, medications, diet, or lifestyle. mined. A rating of 0 is considered normal. A rating of 1.0 is 10 considered to correspond to mild dementia. A Subject with a The methods of the present invention can be used for one or CDR of 0.5 is characterized by mild consistent forgetfulness, more of the following: to prevent Alzheimer's disease, to treat partial recollection of events and “benign forgetfulness. In Alzheimer's disease, or ameliorate symptoms of Alzheimer's one embodiment the Subject is assessed with a rating on the disease, or to regulate production of or levels of amyloid B CDR of above 0, of above about 0.5, of above about 1.0, of (AB) peptides. In an embodiment, the human carries one or 15 above about 1.5, of above about 2.0, of above about 2.5, or at more mutations in the genes that encode 3-amyloid precursor about 3.0. protein, presenilin-1 or presenilin-2. In another embodiment, Another test is the Mini-Mental State Examination the human carries the Apollipoprotein e4 gene. In another (MMSE), as described by Folstein “Mini-mental state. A embodiment, the human has a family history of Alzheimer's practical method for grading the cognitive state of patients for Disease or a dementia illness. In another embodiment, the the clinician.” J. Psychiatr. Res. 12:189-198, 1975. The human has trisomy 21 (Down's Syndrome). In another MMSE evaluates the presence of global intellectual deterio embodiment, the Subject has a normal or low serum total ration. See also Folstein “Differential diagnosis of dementia. blood cholesterol level. In another embodiment, the serum The clinical process.” Psychiatr Clin North Am. 20:45-57, total blood cholesterol level is less than about 200 mg/dL, or 1997. The MMSE is a means to evaluate the onset of dementia less than about 180, and it can range from about 150 to about 25 and the presence of global intellectual deterioration, as seen 200 mg/dL. In another embodiment, the total LDL choles in Alzheimer's disease and multi-infart dementia. The terol level is less than about 100 mg/dL, or less than about 90 MMSE is scored from 1 to 30. The MMSE does not evaluate mg/dL and can range from about 30 to about 100 mg/dL. basic cognitive potential, as, for example, the so-called IQ Methods of measuring serum total blood cholesterol and total test. Instead, it tests intellectual skills. A person of “normal LDL cholesterol are well known to those skilled in the art and 30 intellectual capabilities will score a “30” on the MMSE for example include those disclosed in WO99/38498 at p. 11, objective test (however, a person with a MMSE score of 30 incorporated by reference herein. Methods of determining could also score well below "normal” on an IQ test). See, e.g., levels of other sterols in serum are disclosed in H. Gylling, et Kaufer, J. Neuropsychiatry Clin. Neurosci. 10:55-63, 1998: al., “Serum Sterols During Stanol Ester Feeding in a Mildly Becke, Alzheimer Dis Assoc Disord. 12:54–57, 1998: Ellis, Hypercholesterolemic Population, J. Lipid Res. 40: 593-600 35 Arch. Neurol. 55:360-365, 1998; Magni, Int. Psychogeriatr. (1999). 8:127-134, 1996: Monsch, Acta Neurol. Scand. 92:145-150, In another embodiment, the Subject has an elevated serum 1995. In one embodiment, the subject scores below 30 at least total blood cholesterol level. In another embodiment, the once on the MMSE. In another embodiment, the subject serum total cholesterol level is at least about 200 mg/dL, or at scores below about 28, below about 26, below about 24, least about 220 mg/dL and can range from about 200 to about 40 below about 22, below about 20, below about 18, below about 1000 mg/dL. In another embodiment, the subject has an 16, below about 14, below about 12, below about 10, below elevated total LDL cholesterol level. In another embodiment, about 8, below about 6, below about 4, below about 2, or the total LDL cholesterol level is greater than about 100 below about 1. mg/dL, or even greater than about 110 mg/dL and can range The Disability Assessment for Dementia (“DAD) scale from about 100 to about 1000 mg/dL. 45 has been developed to measure a patient’s ability to perform In another embodiment, the human is at least about 40 the activities of daily living (Gélinas I et al. Development of years of age. In another embodiment, the human is at least a Functional Measure for Persons with Alzheimer's Disease: about 60years of age. In another embodiment, the human is at The Disability Assessment for Dementia. Am. J. Occupa least about 70 years of age. In another embodiment, the tional Therapy. 1999; 53: 471-481). Activities of daily living human is at least about 80 years of age. In another embodi 50 may be assessed according to self care (i.e., dressing and ment, the human is at least about 85 years of age. In one personal hygiene) and instrumental activities (e.g., house embodiment, the human is between about 60 and about 100 work, cooking, and using household devices). The objectives years of age. of the DAD scale include quantitatively measuring functional In still a further embodiment, the subject is shown to be at abilities in activities of daily living in individuals with cog risk by a diagnostic brain imaging technique, for example, 55 nitive impairments and to help delineate areas of cognitive one that measures brain activity, plaque deposition, or brain deficits that may impair performance in activities of daily atrophy. living. The DAD is administered through an interview with In still a further embodiment, the subject is shown to be at the caregiver. It measures actual performance in activities of risk by a cognitive test Such as Clinical Dementia Rating daily living of the individual as observed over a 2 week period (“CDR), Alzheimer's Disease Assessment Scale-Cognition 60 prior to the interview. The scale assesses the following (ADAS-Cog'), Disability Assessment for Dementia domains of activities: hygiene, dressing, telephoning, conti (“DAD) or Mini-Mental State Examination (“MMSE).The nence, eating, meal preparation, outing activities, finance and Subject may exhibit a below average score on a cognitive test, correspondence, medication use, leisure and housework. A as compared to a historical control of similar age and educa total score is obtained by adding the rating for each question tional background. The Subject may also exhibit a reduction 65 and converting this total score out of 100. Higher scores in score as compared to previous scores of the Subject on the represent less disability in ADL while lower scores indicate same or similar cognition tests. more dysfunction. In one embodiment, the Subject scores US 8,642,801 B2 175 176 below 100 at least once on the DAD. In another embodiment, AB/mL, or greater than about 400 pg/mL. In another the subject scores below about 95, below about 90, below embodiment, the elevated level of amyloid Afa peptide can about 85, below about 80, below about 75, below about 70, range from about 200 pg/mL to about 800 pg/mL, to even below about 65, below about 60, below about 55, below about about 1000 pg/mL. 50, below about 45, below about 40, below about 30, below In another embodiment, the Subject can have an elevated about 20, or below about 10. level of amyloid AB peptide in the CSF prior to treatment, Another means to evaluate cognition, particularly Alzhe according to the present methods, of greater than about 10 imer's disease, is the Alzheimer's Disease Assessment Scale pg/mL, or greater than about 10 pg/mL, or greater than about (ADAS-Cog), or a variation termed the Standardized Alzhe 200 pg/mL, or greater than about 500 pg/mL. In another imer's Disease Assessment Scale (SADAS). It is commonly 10 embodiment, the level of amyloid B peptide can range from used as an efficacy measure in clinical drug trials of Alzhe about 10 pg/mL to about 1,000 pg/mL, or even about 100 imer's disease and related disorders characterized by cogni pg/mL to about 1,000 pg/mL. tive decline. SADAS and ADAS-Cog were not designed to In another embodiment, the Subject can have an elevated diagnose Alzheimer's disease; they are useful in characteriz level of amyloid AB peptide in the CSF prior to treatment ing symptoms of dementia and are a relatively sensitive indi 15 according to the present methods of greater than about cator of dementia progression. (See, e.g., Doraiswamy, Neu pg/mL, or greater than about 50 pg/mL, or even greater than rology 48:1511-1517, 1997; and Standish, J. Am. Geriatr. about 100 pg/mL. In another embodiment, the level of amy Soc. 44:712-716, 1996.) Annual deterioration in untreated loid f peptide can range from about 10 pg/mL to about 1,000 Alzheimer's disease patients is approximately 8 points per pg/mL. year (See, e.g., Raskind, M. Prim. Care CompanionJ Clin The amount of amyloid f peptide in the brain, CSF, blood, Psychiatry 2000 August; 2(4): 134-138). or plasma of a Subject can be evaluated by enzyme-linked The ADAS-cog is designed to measure, with the use of immunosorbent assay (“ELISA) or quantitative immunob questionnaires, the progression and the severity of cognitive lotting test methods or by quantitative SELDI-TOF which are decline as seen in AD on a 70-point scale. The ADAS-cog well known to those skilled in the art, such as is disclosed by scale quantifies the number of wrong answers. Consequently, 25 Zhang, et al., J. Biol. Chem. 274,8966-72 (1999) and Zhang, a high score on the scale indicates a more severe case of et al., Biochemistry 40, 5049-55 (2001). See also, A. K. Veh cognitive decline. In one embodiment, a Subject exhibits a mas, et al., DNA Cell Biol. 20(11), 713-21 (2001), P. Lewc score of greater than 0, greater than about 5, greater than Zuk, et al., Rapid Commun. Mass Spectrom. 17(12), 1291-96 about 10, greater than about 15, greater than about 20, greater (2003); B. M. Austen, et al., J. Peptide Sci. 6, 459-69 (2000); than about 25, greater than about 30, greater than about 35, 30 and H. Davies, et al., BioTechniques 27, 1258-62 (1999). greater than about 40, greater than about 45, greater than These tests are performed on samples of the brain or blood about 50, greater than about 55, greater than about 60, greater which have been prepared in a manner well known to one than about 65, greater than about 68, or about 70. In another skilled in the art. Another example of a useful method for embodiment, the subject exhibits no symptoms of Alzhe measuring levels of amyloid B peptides is by Europium imer's Disease. In another embodiment, the subject is a 35 immunoassay (EIA). See, e.g., WO 99/38498 at p. 11. human who is at least 40 years of age and exhibits no symp The methods of the invention may be applied as a therapy toms of Alzheimer's Disease. In another embodiment, the for a Subject having Alzheimer's disease or a dementia, or the Subject is a human who is at least 40 years of age and exhibits methods of the invention may be applied as a prophylaxis one or more symptoms of Alzheimer's Disease. against Alzheimer's disease or dementia for Subject with Such In another embodiment, the subject has Mild Cognitive 40 a predisposition, as in a Subject, e.g., with a genomic mutation Impairment. In a further embodiment, the subject has a CDR in the APP gene, the ApoE gene, or a presenilin gene. The rating of about 0.5. In another embodiment, the subject has Subject may have (or may be predisposed to developing or early Alzheimer's disease. In another embodiment, the sub may be suspected of having) vascular dementia, or senile ject has cerebral amyloid angiopathy. dementia, Mild Cognitive Impairment, or early Alzheimer's By using the methods of the present invention, the levels of 45 disease. In addition to Alzheimer's disease, the Subject may amyloid B peptides in a subjects plasma or cerebrospinal have another amyloid-related disease Such as cerebral amy fluid (CSF) can be reduced from levels prior to treatment from loid angiopathy, or the Subject may have amyloid deposits, about 10 to about 100 percent, or even about 50 to about 100 especially amyloid-f amyloid deposits in the brain. percent. Treatment of Amyloid-Related Diseases In an alternative embodiment, the Subject can have an 50 The present invention pertains to methods of using the elevated level of amyloid AB and AB peptide in the blood compounds and pharmaceutical compositions thereof in the and CSF prior to treatment, according to the present methods, treatment and prevention of amyloid-related diseases. The of greater than about 10 pg/mL, or greater than about 20 pharmaceutical compositions of the invention may be admin pg/mL, or greater than about 35 pg/mL, or even greater than istered therapeutically or prophylactically to treat diseases about 40 pg/mL. In another embodiment, the elevated level of 55 associated with amyloid (e.g., AL amyloid protein (w or amyloid AB peptide can range from about 30 pg/mL to K-chain related, e.g., amyloid W., amyloid K, amyloid KIV. about 200 pg/mL, or even to about 500 pg/mL. One skilled in amyloid VI, amyloidy, amyloid Y1), AB, IAPP. B.M., AA, or the art would understand that as Alzheimer's disease AHamyloid protein) fibril formation, aggregation or deposi progresses, the measurable levels of amyloid B peptide in the tion. CSF may decrease from elevated levels present before onset 60 The pharmaceutical compositions of the invention may act of the disease. This effect is attributed to increased deposi to ameliorate the course of an amyloid-related disease using tion, i.e., trapping of AB peptide in the brain instead of normal any of the following mechanisms (this list is meant to be clearance from the brain into the CSF. illustrative and not limiting): slowing the rate of amyloid fibril In an alternative embodiment, the Subject can have an formation or deposition; lessening the degree of amyloid elevated level of amyloid ABao peptide in the blood and CSF 65 deposition; inhibiting, reducing, or preventing amyloid fibril prior to treatment, according to the present methods, of formation; inhibiting neurodegeneration or cellular toxicity greater than about 5pg AB/mL or greater than about 50 pg induced by amyloid; inhibiting amyloid induced inflamma US 8,642,801 B2 177 178 tion; enhancing the clearance of amyloid from the brain; tion Successfully treat a Subject's dementia by slowing the enhancing degradation of AB in the brain; or favoring clear rate of or lessening the extent of cognitive decline. ance of amyloid protein prior to its organization in fibrils. In one embodiment, the term “treating includes maintain “Modulation of amyloid deposition includes both inhibi ing a subject's CDR rating at its base line rating or at 0. In tion, as defined above, and enhancement of amyloid deposi another embodiment, the term treating includes decreasing a tion or fibril formation. The term “modulating is intended, subject's CDR rating by about 0.25 or more, about 0.5 or therefore, to encompass prevention or stopping of amyloid more, about 1.0 or more, about 1.5 or more, about 2.0 or more, formation or accumulation, inhibition or slowing down of about 2.5 or more, or about 3.0 or more. In another embodi further amyloid formation or accumulation in a subject with ment, the term “treating also includes reducing the rate of the ongoing amyloidosis, e.g., already having amyloid deposi 10 increase of a subject's CDR rating as compared to historical controls. In another embodiment, the term includes reducing tion, and reducing or reversing of amyloid formation or accu the rate of increase of a subjects CDR rating by about 5% or mulation in a Subject with ongoing amyloidosis; and enhanc more, about 10% or more, about 20% or more, about 25% or ing amyloid deposition, e.g., increasing the rate or amount of more, about 30% or more, about 40% or more, about 50% or amyloid deposition in vivo or in vitro. Amyloid-enhancing 15 more, about 60% or more, about 70% or more, about 80% or compounds may be useful in animal models of amyloidosis, more, about 90% or more, or about 100%, of the increase of for example, to make possible the development of amyloid the historical or untreated controls. deposits in animals in a shorter period of time or to increase In another embodiment, the term “treating also includes amyloid deposits over a selected period of time. Amyloid maintaining a subjects score on the MMSE. The term “treat enhancing compounds may be useful in screening assays for ing includes increasing a subject’s MMSE score by about 1, compounds which inhibit amyloidosis in Vivo, for example, about 2, about 3, about 4, about 5, about 7.5, about 10, about in animal models, cellular assays and in vitro assays for 12.5, about 15, about 17.5, about 20, or about 25 points. The amyloidosis. Such compounds may be used, for example, to term also includes reducing the rate of the decrease of a provide faster or more sensitive assays for compounds. subject's MMSE score as compared to historical controls. In Modulation of amyloid deposition is determined relative to an 25 another embodiment, the term includes reducing the rate of untreated subject or relative to the treated subject prior to decrease of a subjects MMSE score by about 5% or less, treatment. about 10% or less, about 20% or less, about 25% or less, about “Inhibition of amyloid deposition includes preventing or 30% or less, about 40% or less, about 50% or less, about 60% stopping of amyloid formation, e.g., fibrillogenesis, clear or less, about 70% or less, about 80% or less, about 90% or ance of amyloid, e.g., Soluble AB from brain, inhibiting or 30 less or about 100% or less, of the decrease of the historical or slowing down of further amyloid deposition in a subject with untreated controls. amyloidosis, e.g., already having amyloid deposits, and In another embodiment, the term “treating also includes reducing or reversing amyloid fibrillogenesis or deposits in a maintaining a subject's score on the DAD. The term “treat Subject with ongoing amyloidosis. Inhibition of amyloid ing includes increasing a subject’s DAD score by about 1, deposition is determined relative to an untreated Subject, or 35 about 5, about 10, about 15, about 20, about 30, about 35, relative to the treated Subject prior to treatment, or, e.g., about 40, about 50, about 60, about 70, or about 80 points. The determined by clinically measurable improvement, e.g., or in term also includes reducing the rate of the decrease of a the case of a Subject with brain amyloidosis, e.g., an Alzhe subject’s DAD score as compared to historical controls. In imer's or cerebral amyloid angiopathy Subject, stabilization another embodiment, the term includes reducing the rate of of cognitive function or prevention of a further decrease in 40 decrease of a subject’s DAD score by about 5% or less, about cognitive function (i.e., preventing, slowing, or stopping dis 10% or less, about 20% or less, about 25% or less, about 30% ease progression), or improvement of parameters such as the or less, about 40% or less, about 50% or less, about 60% or concentration of AB or tau in the CSF. less, about 70% or less, about 80% or less, about 90% or less As used herein, “treatment of a subject includes the appli or about 100% or less, of the decrease of the historical or cation or administration of a composition of the invention to 45 untreated controls. a subject, or application or administration of a composition of In yet another embodiment, the term “treating includes the invention to a cell or tissue from a Subject, who has an maintaining a subjects score on the ADAS-Cog. The term amyloid-related disease or condition, has a symptom of Such “treating includes decreasing a Subject's ADAS-Cog score a disease or condition, or is at risk of (or Susceptible to) Such by about 1 point or greater, by about 2 points or greater, by a disease or condition, with the purpose of curing, healing, 50 about 3 points or greater, by about 4 points or greater, by about alleviating, relieving, altering, remedying, ameliorating, 5 points or greater, by about 7.5 points or greater, by about 10 improving, or affecting the disease or condition, the symptom points or greater, by about 12.5 points or greater, by about 15 of the disease or condition, or the risk of (or susceptibility to) points or greater, by about 17.5 points or greater, by about 20 the disease or condition. The term “treating refers to any points or greater, or by about 25 points or greater. The term indicia of Success in the treatment or amelioration of an 55 also includes reducing the rate of the increase of a subjects injury, pathology or condition, including any objective or ADAS-Cog score as compared to historical controls. In Subjective parameter Such as abatement; remission; diminish another embodiment, the term includes reducing the rate of ing of symptoms or making the injury, pathology or condition increase of a subject’s ADAS-Cog score by about 5% or more tolerable to the Subject; slowing in the rate of degenera more, about 10% or more, about 20% or more, about 25% or tion or decline; making the final point of degeneration less 60 more, about 30% or more, about 40% or more, about 50% or debilitating; improving a subject’s physical or mental well more, about 60% or more, about 70% or more, about 80% or being; or, in Some situations, preventing the onset of demen more, about 90% or more or about 100% of the increase of the tia. The treatment or amelioration of symptoms can be based historical or untreated controls. on objective or subjective parameters; including the results of In another embodiment, the term “treating” e.g., for AA or a physical examination, a psychiatric evaluation, or a cogni 65 AL amyloidosis, includes an increase in serum creatinine, tion test such as CDR, MMSE, DAD, ADAS-Cog, or another e.g., an increase of creatinine clearance of 10% or greater, test known in the art. For example, the methods of the inven 20% or greater, 50% or greater, 80% or greater, 90% or US 8,642,801 B2 179 180 greater, 100% or greater, 150% or greater, 200% or greater. neurodegeneration, and it has been shown that neuronal cells The term “treating also may induce remission of nephrotic exposed in vitro to AB can become apoptotic (Morgan et al. syndrome (NS). It may also include remission of chronic (2004) Prog. Neurobiol. 74:323-349; Stefani et al. (2003).J. diarrhea and/or again in body weight, e.g., by 10% or greater, Mol. Med. 81:678-99; La Ferla et al. (1997).J. Clin. Invest. 15% or greater, or 20% or greater. 100(2):310-320). In Alzheimer's disease, a progressive neu Without wishing to be bound by theory, in some aspects the ronal cell loss accompanies the deposition of AB amyloid pharmaceutical compositions of the invention contain a com fibrils in senile plaques. pound that prevents or inhibits amyloid fibril formation, In yet another aspect, the invention pertains to a method for either in the brain or other organ of interest (acting locally) or inhibiting AB-induced neuronal cell death by administering throughout the entire body (acting systemically). Pharmaceu 10 an effective amount of a compound of the present invention. tical compositions of the invention may be effective in con Another aspect of the invention pertains to a method for trolling amyloid deposition either following their entry into providing neuroprotection to a Subject having an AB-amyloid the brain (following penetration of the blood brain bather) or related disease, e.g., Alzheimer's disease, that includes from the periphery. When acting from the periphery, a com administering an effective amount of a compound of the pound of a pharmaceutical composition may alter the equi 15 present invention to the Subject, Such that neuroprotection is librium of amyloidogenic peptide between the brain and the provided. plasma So as to favor the exit of amyloidogenic peptide from In another aspect, methods for inhibiting AB-induced neu the brain. It may also favor clearance (or catabolism) of the ronal cell death are provided that include administration of an amyloid protein (soluble), and then prevent amyloid fibril effective amount of a compound of the present invention to a formation and deposition due to a reduction of the amyloid subject such that neuronal cell death is inhibited. protein pool in a specific organ, e.g., liver, spleen, pancreas, In another aspect, methods for treating a disease state char kidney, joints, brain, etc. An increase in the exit of amy acterized by AB-induced neuronal cell death in a subject are loidogenic peptide from the brain would result in a decrease provided, e.g., by administering an effective amount of a in amyloidogenic peptide brain concentration and therefore compound of the present invention. Non-limiting examples of favor a decrease in amyloidogenic peptide deposition. In 25 Such disease states include Alzheimer's disease and AB-amy particular, an agent may lower the levels of amyloid B pep loid related diseases. tides, e.g., both A?40 and AB42 in the CSF and the plasma, or The term “neuroprotection includes protection of neu the agent may lower the levels of amyloid B peptides, e.g., AB ronal cells of a subject from AB-induced cell death, e.g., cell 40 and AB42 in the CSF and increase it in the plasma. Alter death induced directly or indirectly by an AB peptide. Af natively, compounds that penetrate the brain could control 30 induced cell death may result in initiation of processes Such deposition by acting directly on brain amyloidogenic peptide as, for example: the destabilization of the cytoskeleton; DNA e.g., by maintaining it in a non-fibrillar form or favoring its fragmentation; the activation of hydrolytic enzymes, such as clearance from the brain, by increasing its degradation in the phospholipase A2; activation of caspases, calcium-activated brain, or protecting brain cells from the detrimental effect of proteases and/or calcium-activated endonucleases; inflam amyloidogenic peptide. An agent can also cause a decrease of 35 mation mediated by macrophages; calcium influx into a cell; the concentration of the amyloid protein (i.e., in a specific membrane potential changes in a cell; the disruption of cell organ so that the critical concentration needed to trigger amy junctions leading to decreased or absent cell-cell communi loid fibril formation or deposition is not reached). Further cation; and the activation of expression of genes involved in more, the compounds described herein may inhibit or reduce cell death, e.g., immediate-early genes. an interaction between amyloid and a cell Surface constituent, 40 The term "amyloid-fi disease' (or "amyloid? related dis for example, a glycosaminoglycan or proteoglycan constitu ease, which terms as used herein are synonymous) may be ent of a basement membrane, whereby inhibiting or reducing used for mild cognitive impairment; vascular dementia; early this interaction produces the observed neuroprotective and Alzheimer's disease; Alzheimer's disease, including spo cell-protective effects. For example, the compound may also radic (non-hereditary) Alzheimer's disease and familial (he prevent an amyloid peptide from binding or adhering to a cell 45 reditary) Alzheimer's disease; age-related cognitive decline; Surface, a process which is known to cause cell damage or cerebral amyloid angiopathy (“CAA'); hereditary cerebral toxicity. Similarly, the compound may block amyloid-in hemorrhage; senile dementia; Down's syndrome; inclusion duced cellular toxicity or microglial activation or amyloid body myositis (“IBM’); or age-related macular degeneration induced neurotoxicity, or inhibit amyloid induced inflamma (ARMD). According to certain aspects of the invention, tion. The compound may also reduce the rate or amount of 50 amyloid-fi is a peptide having 39-43 amino-acids, or amy amyloid aggregation, fibril formation, or deposition, or the loid-fi is an amyloidogenic peptide produced from BAPP. compound lessens the degree of amyloid deposition. The Mild cognitive impairment (MCI) is a condition charac foregoing mechanisms of action should not be construed as terized by a state of mild but measurable impairment in think limiting the scope of the invention inasmuch as the invention ing skills, which is not necessarily associated with the pres may be practiced without Such information. 55 ence of dementia. MCI frequently, but not necessarily, The AB peptide has been shown by several groups to be precedes Alzheimer's disease. It is a diagnosis that has most highly toxic to neurons. Amyloid plaques are directly associ often been associated with mild memory problems, but it can ated with reactive gliosis, dystrophic neurites and apoptotic also be characterized by mild impairments in other thinking cells, suggesting that plaques induce neurodegenerative skills, such as language or planning skills. However, in gen changes. Neurotoxicity may eventually disrupt or even kill 60 eral, an individual with MCI will have more significant neurons. In vitro, AB has been shown to induce apoptosis in memory lapses than would be expected for someone of their many different neuronal cell types, such as rat PC-12 cells, age or educational background. As the condition progresses, primary rathippocampal and cortical cultures, and the pred a physician may change the diagnosis to "Mild-to-Moderate ifferentiated human neurotype SH-SY5Y cell line (Dickson Cognitive Impairment, as is well understood in this art. DW (2004).J Clin Invest 114:23-7: Canu et al. (2003) Cer 65 Cerebral amyloid angiopathy (“CAA) refers to the spe ebellum 2:270-278: Lietal. (1996) Brain Research 738: 196 cific deposition of amyloid fibrils in the walls of leptomingeal 204). Numerous reports have shown that AB fibrils can induce and cortical arteries, arterioles and in capillaries and veins. It US 8,642,801 B2 181 182 is commonly associated with Alzheimer's disease, Down's tering a compound capable of reducing the concentration of syndrome and normal aging, as well as with a variety of amyloid or reducing the interaction of an amyloid with a cell familial conditions related to stroke or dementia (see Fran Surface. gione, et al., Amyloid. J. Protein Folding Disord. 8, Suppl. 1, The invention also includes a method for directly or indi 36-42 (2001)). CAA can occur sporadically or be hereditary. rectly preventing cell death in a subject, the method compris Multiple mutation sites in either AB or the APP gene have ing administering to a Subject a therapeutically effective been identified and are clinically associated with either amount of a compound capable of preventing amyloid (e.g., dementia or cerebral hemorrhage. Exemplary CAA disorders AL amyloid protein (W or K-chain related, e.g., amyloid W. include, but are not limited to, hereditary cerebral hemor amyloid K, amyloid KIV, amyloid wVI, amyloid Y, amyloid 10 Y1), AB, IAPP. Bam, AA, AH amyloid protein, or other amy rhage with amyloidosis of Icelandic type (HCHWA-I); the loid) mediated events that lead, directly or indirectly, to cell Dutch variant of HCHWA (HCHWA-D; a mutation in AB); death. the Flemish mutation of AB; the Arctic mutation of AB; the In an embodiment, the method is used to treat Alzheimer's Italian mutation of AB; the Iowa mutation of AB; familial disease (e.g. sporadic or familial AD). The method can also be British dementia; and familial Danish dementia. Cerebral 15 used prophylactically or therapeutically to treat other clinical amyloid angiopathy is known to be associated with cerebral occurrences of amyloid-fi deposition, such as in Down's Syn hemorrhage (or hemorrhagic stroke). drome individuals and in patients with cerebral amyloid angi Additionally, abnormal accumulation of APP and of amy opathy (“CAA') or hereditary cerebral hemorrhage. loid-fi protein in muscle fibers has been implicated in the The compounds of the invention may be used prophylac pathology of sporadic inclusion body myositis (“IBM) tically or therapeutically in the treatment of disorders in (Askanas, et al., Proc. Natl. Acad. Sci. USA 93, 1314-19 which amyloid-beta peptide is abnormally deposited at non (1996); Askanas, et al., Current Opinion in Rheumatology 7. neurological locations, such as treatment of IBM by delivery 486-96 (1995)). Accordingly, the compounds of the invention of the compounds to muscle fibers, or treatment of macular can be used prophylactically or therapeutically in the treat degeneration by delivery of the compound(s) of the invention ment of disorders in which amyloid-fi protein is abnormally 25 to the basal surface of the retinal pigmented epithelium. deposited at non-neurological locations, such as treatment of The present invention also provides a method for modulat IBM by delivery of the compounds to muscle fibers. ing amyloid-associated damage to cells, comprising the step Additionally, it has been shown that AB is associated with of administering a compound capable of reducing the con abnormal extracellular deposits, known as drusen, that accu centration of AB, or capable of minimizing the interaction of mulate along the basal Surface of the retinal pigmented epi 30 A? (soluble oligomeric or fibrillary) with the cell surface, thelium in individuals with age-related macular degeneration Such that said amyloid-associated damage to cells is modu (ARMD). ARMD is a cause of irreversiblevision loss in older lated. In certain aspects of the invention, the methods for individuals. It is believed that AB deposition could be an modulating amyloid-associated damage to cells comprise a important component of the local inflammatory events that step of administering a compound capable of reducing the contribute to atrophy of the retinal pigmented epithelium, 35 concentration of AB or reducing the interaction of AB with a drusen biogenesis, and the pathogenesis of ARMD (Johnson, cell surface. et al., Proc. Natl. Acad. Sci. USA 99(18), 11830-5 (2002)). In accordance with the present invention, there is further Therefore, the invention also relates to the treatment or pre provided a method for preventing cell death in a Subject, said vention of age-related macular degeneration. method comprising administering to a subject a therapeuti Also, the invention relates to a method for preventing or 40 cally effective amount of a compound capable of preventing inhibiting amyloid deposition in a Subject. For example. Such Af-mediated events that lead, directly or indirectly, to cell a method comprises administering to a subject a therapeuti death. cally effective amount of a compound capable of reducing the The present invention also provides a method for modulat concentration of amyloid (e.g., AL amyloid protein (w or ing amyloid-associated damage to cells, comprising the step K-chain related, e.g., amyloid W., amyloid K, amyloid KIV. 45 of administering a compound capable of reducing the con amyloid wVI, amyloid Y, amyloid Y1), AB, IAPP. Bam, AA, centration of IAPP, or capable of minimizing the interaction AH amyloid protein, or other amyloids). Such that amyloid of IAPP (soluble oligomeric or fibrillary) with the cell sur accumulation or deposition is prevented or inhibited. face, such that said amyloid-associated damage to cells is In another aspect, the invention relates to a method for modulated. In certain aspects of the invention, the methods preventing, reducing, or inhibiting amyloid deposition in a 50 for modulating amyloid-associated damage to cells comprise Subject. For example, such a method comprises administering a step of administering a compound capable of reducing the to a subject atherapeutically effective amount of a compound concentration of IAPP or reducing the interaction of IAPP capable of inhibiting amyloid (e.g., AL amyloid protein (w or with a cell surface. K-chain related, e.g., amyloid W., amyloid K, amyloid KIV. In accordance with the present invention, there is further amyloid wVI, amyloid Y, amyloid Y1), AB, IAPP. Bam, AA, 55 provided a method for preventing cell death in a Subject, said AH amyloid protein, or other amyloids). Such that amyloid method comprising administering to a subject a therapeuti deposition is prevented, reduced, or inhibited. cally effective amount of a compound capable of preventing The invention also relates to a method for modulating, e.g., IAPP-mediated events that lead, directly or indirectly, to cell minimizing, amyloid-associated damage to cells, comprising death. the step of administering a compound capable of reducing the 60 This invention also provides methods and compositions concentration of amyloid (e.g., AL amyloid protein (w or which are useful in the treatment of amyloidosis. The meth K-chain related, e.g., amyloid W., amyloid K, amyloid KIV. ods of the invention involve administering to a subject a amyloid VI, amyloidy, amyloid Y1), Af, IAPP. B.m, AA, therapeutic compound which inhibits amyloid deposition. AH amyloid protein, or another amyloid). Such that said Accordingly, the compositions and methods of the invention amyloid-associated damage to cells is modulated. In certain 65 are useful for inhibiting amyloidosis in disorders in which aspects of the invention, the methods for modulating amy amyloid deposition occurs. The methods of the invention can loid-associated damage to cells comprise a step of adminis be used therapeutically to treat amyloidosis or can be used US 8,642,801 B2 183 184 prophylactically in a Subject Susceptible to (hereditary) amy absorption assay. In an exemplary protocol, a test compound loidosis or identified as being at risk to develop amyloidosis, (20 uM) is incubated with 50 uMAB(1-40) fibers for 1 hour at e.g., hereditary, or identified as being at risk to develop amy 37°C. in Tris buffered saline (20 mM Tris, 150 mMNaCl, pH loidosis. In certain embodiments, the invention includes a 7.4 containing 0.01 sodium azide). Following incubation, the method of inhibiting an interaction between an amy solution is centrifuged for 20 minutes at 21,000 g to sediment loidogenic protein and a constituent of basement membrane the AB(1-40) fibers along with any bound test compound. The to inhibit amyloid deposition. The constituent of basement amount of test compound remaining in the Supernatant can membrane is a glycoprotein or proteoglycan, preferably then be determined by reading the absorbance. The fraction of heparan Sulfate proteoglycan. A therapeutic compound used test compound bound can then be calculated by comparing in this method may interfere with binding of a basement 10 membrane constituent to a target binding site on an amy the amount remaining in the Supernatants of incubations with loidogenic protein, thereby inhibiting amyloid deposition. AB to the amount remaining in control incubations which do In some aspects, the methods of the invention involve not contain AB fibers. Thioflavin T and Congo Red, both of administering to a subject a therapeutic compound which which are known to bind to AB fibers, may be included in each inhibits amyloid deposition “Inhibition of amyloid deposi 15 assay run as positive controls. Before assaying, test com tion, includes the prevention of amyloid formation, inhibi pounds can be diluted to 40 uM, which would be twice the tion of further amyloid deposition in a Subject with ongoing concentration in the final test, and then Scanned using the amyloidosis and reduction of amyloid deposits in a subject Hewlett Packard 8453 UV/VIS spectrophotometer to deter with ongoing amyloidosis Inhibition of amyloid deposition is mine if the absorbance is sufficient for detection. determined relative to an untreated subject or relative to the In another embodiment, the invention pertains to a method treated Subject prior to treatment. In an embodiment, amyloid for improving cognition in a Subject Suffering from an amy deposition is inhibited by inhibiting an interaction between an loid-related disease. The method includes administering an amyloidogenic protein and a constituent of basement mem effective amount of a therapeutic compound of the invention, brane. “Basement membrane' refers to an extracellular Such that the Subjects cognition is improved. The Subjects matrix comprising glycoproteins and proteoglycans, includ 25 cognition can be tested using methods known in the art Such ing laminin, collagen type IV, fibronectin, perlecan, agrin, as the Clinical Dementia Rating (“CDR), Mini-Mental State dermatan sulfate, and heparan sulfate proteoglycan (HSPG). Examination (“MMSE), Disability Assessment for Demen In one embodiment, amyloid deposition is inhibited by inter tia (“DAD), and/or the Alzheimer's Disease Assessment fering with an interaction between an amyloidogenic protein Scale-Cognition (ADAS-Cog'). and a sulfated glycosaminoglycan such as HSPG, dermatan 30 In another embodiment, the invention pertains to a method Sulfate, perlecan or agrin Sulfate. Sulfated glycosaminogly for treating a subject for an amyloid-related disease. The cans are known to be present in all types of amyloids (see method includes administering a cognitive test to a subject Snow, et al. Lab. Invest. 56, 120-23 (1987)) and amyloid prior to administration of a compound of the invention, deposition and HSPG deposition occur coincidentally in ani administering an effective amount of a compound of the mal models of amyloidosis (see Snow, et al. Lab. Invest. 56, 35 invention to the Subject, and administering a cognitive test to 665-75 (1987) and Gervais, F. et al. Curr. Med. Chem., 3, the Subject Subsequent to administration of the compound, 361-370 (2003)). Consensus binding site motifs for HSPG in such that the subject is treated for the amyloid-related disease, amyloidogenic proteins have been described (see, e.g., Car wherein the Subject's score on said cognitive test is improved. din and Weintraub Arteriosclerosis 9, 21-32 (1989)). “Improvement,” “improved' or “improving in cognition The ability of a compound to prevent or block the forma 40 is present within the context of the present invention if there tion or deposition of amyloid may reside in its ability to bind is a statistically significant difference in the direction of nor to non-fibrillar, soluble amyloid protein and to maintain its mality between the performance of subjects treated using the solubility. methods of the invention as compared to members of a pla The ability of a therapeutic compound of the invention to cebo group, historical control, or between Subsequent tests inhibit an interaction between an amyloidogenic protein and 45 given to the same Subject. a glycoprotein or proteoglycan constituent of a basement In one embodiment, a subject's CDR is maintained at 0. In membrane can be assessed by an in vitro binding assay, Such another embodiment, a subject's CDR is decreased (e.g., as that described in U.S. Pat. No. 5,164,295, the contents of improved) by about 0.25 or more, about 0.5 or more, about 1.0 which are hereby incorporated by reference. Alternatively, or more, about 1.5 or more, about 2.0 or more, about 2.5 or the ability of a compound to bind to an amyloidogenic protein 50 more, or about 3.0 or more. In another embodiment, the rate or to inhibit the binding of a basement membrane constituent of increase of a subject's CDR rating is reduced by about 5% (e.g., HSPG) to an amyloidogenic protein (e.g., AB) can be or more, about 10% or more, about 20% or more, about 25% measured using a mass spectrometry assay where soluble or more, about 30% or more, about 40% or more, about 50% protein, e.g., Af, IAPP. B.M is incubated with the compound. or more, about 60% or more, about 70% or more, about 80% A compound which binds to, e.g., AB, will induce a change in 55 or more, about 90% or more, or about 100% or more of the the mass spectrum of the protein. Exemplary protocols for a increase of the historical or untreated controls. mass spectrometry assay employing AB and IAPP can be In one embodiment, a subjects score on the MMSE is found in the Examples, the results of which are provided in maintained. Alternatively, the subjects score on the MMSE Table 3. The protocol can readily be modified to adjust the may be increased by about 1, about 2, about 3, about 4, about sensitivity of the data, e.g., by adjusting the amount of protein 60 5, about 7.5, about 10, about 12.5, about 15, about 17.5, about and/or compound employed. Thus, e.g., binding might be 20, or about 25 points. In another alternative, the rate of the detected for test compounds noted as not having detectable decrease of a subject's MMSE score as compared to historical binding employing less sensitive test protocols. controls is reduced. For example, the rate of the decrease of a Alternative methods for Screening compounds exist and subject's MMSE score may be reduced by about 5% or more, can readily be employed by a skilled practitioner to provide 65 about 10% or more, about 20% or more, about 25% or more, an indication of the ability of test compounds to bind to, e.g., about 30% or more, about 40% or more, about 50% or more, fibrillar AB. One Such screening assay is an ultraviolet about 60% or more, about 70% or more, about 80% or more, US 8,642,801 B2 185 186 about 90% or more, or about 100% or more of the decrease of Synthesis of Compounds of the Invention the historical or untreated controls. In general, the compounds of the present invention may be In one embodiment, a subject’s score on the DAD is main prepared by the methods illustrated in the general reaction tained. Alternatively, the subject's score on the DAD may be schemes as, for example, described below, or by modifica increased by about 1, about 2, about 3, about 4, about 5, about tions thereof, using readily available starting materials, 7.5, about 10, about 15, about 20, about 30, about 40, or about reagents and conventional synthesis procedures. In these 50 or more points. In another alternative, the rate of the reactions, it is also possible to make use of variants which are decrease of a subject’s DAD score as compared to historical in themselves known, but are not mentioned here. Functional controls is reduced. For example, the rate of the decrease of a and structural equivalents of the compounds described herein 10 and which have the same general properties, wherein one or subject’s DAD score may be reduced by about 5% or more, more simple variations of Substituents are made which do not about 10% or more, about 20% or more, about 25% or more, adversely affect the essential nature or the utility of the com about 30% or more, about 40% or more, about 50% or more, pound are also included. about 60% or more, about 70% or more, about 80% or more, The compounds of the present invention may be readily about 90% or more, or about 100% or more of the decrease of 15 prepared in accordance with the synthesis schemes and pro the historical or untreated controls. tocols described herein, as illustrated in the specific proce In one embodiment, the invention pertains to a method for dures provided. However, those skilled in the art will recog treating, slowing or stoppinganamyloid-related disease asso nize that other synthetic pathways for forming the ciated with cognitive impairment, by administering to a Sub compounds of this invention may be used, and that the fol ject an effective amount of a therapeutic compound of the lowing is provided merely by way of example, and is not invention, wherein the annual deterioration of the subjects limiting to the present invention. See, e.g., "Comprehensive cognition as measured by ADAS-Cog is less than 8 points per Organic Transformations” by R. Larock, VCH Publishers year, less the 6 points per year, less than 5 points per year, less (1989). It will be further recognized that various protecting than 4 points per year, or less than 3 points per year. In a and deprotecting strategies will be employed that are standard further embodiment, the invention pertains to a method for 25 in the art (See, e.g., “Protective Groups in Organic Synthesis' treating, slowing or stoppinganamyloid-related disease asso by Greene and Wuts). Those skilled in the relevant arts will ciated with cognition by administering an effective amount of recognize that the selection of any particular protecting group a therapeutic compound of the invention Such that the Sub (e.g., amine and carboxyl protecting groups) will depend on jects cognition as measured by ADAS-Cog remains constant the stability of the protected moiety with regards to the sub over a year. “Constant includes fluctuations of no more than 30 sequent reaction conditions and will understand the appropri 2 points. Remaining constant includes fluctuations of two ate selections. points or less in either direction. In a further embodiment, the Further illustrating the knowledge of those skilled in the art Subjects cognition improves by 2 points or greater per year, is the following sampling of the extensive chemical literature: 3 points or greater per year, 4 point or greater per year, 5 “Chemistry of the Amino Acids” by J.P. Greenstein and M. points or greater per year, 6 points or greater per year, 7 points 35 Winitz, John Wiley & Sons, Inc., New York (1961): “Com or greater per year, 8 points or greater per year, etc. as mea prehensive Organic Transformations” by R. Larock, VCH sured by the ADAS-Cog. In another alternative, the rate of the Publishers (1989); T. D. Ocain, et al., J. Med. Chem. 31, increase of a Subject's ADAS-Cog score as compared to his 2193-99 (1988); E. M. Gordon, et al., J. Med. Chem. 31, torical controls is reduced. For example, the rate of the 2199-10 (1988): “Practice of Peptide Synthesis” by M. increase of a subject’s ADAS-Cog score may be reduced by 40 Bodansky and A. Bodanszky, Springer-Verlag, New York about 5% or more, about 10% or more, about 20% or more, (1984); “Protective Groups in Organic Synthesis” by T. about 25% or more, about 30% or more, about 40% or more, Greene and P. Wuts (1991); 'Asymmetric Synthesis: Con about 50% or more, about 60% or more, about 70% or more, struction of Chiral Molecules Using Amino Acids” by G. M. about 80% or more, about 90% or more or about 100% of the Coppola and H. F. Schuster, John Wiley & Sons, Inc., New increase of the historical or untreated controls. 45 York (1987); “The Chemical Synthesis of Peptides” by J. In another embodiment, the ratio of AB42:AB40 in the CSF Jones, Oxford University Press, New York (1991); and “Intro or plasma of a subject decreases by about 15% or more, about duction of Peptide Chemistry” by P. D. Bailey, John Wiley & 20% or more, about 25% or more, about 30% or more, about Sons, Inc., New York (1992). 35% or more, about 40% or more, about 45% or more, or The synthesis of compounds of the invention is carried out about 50% or more. In another embodiment, the levels of AB 50 in a solvent. Suitable solvents are liquids at ambient room in the subject’s cerebrospinal fluid decrease by about 15% or temperature and pressure or remain in the liquid State under more, about 25% or more, about 35% or more, about 45% or the temperature and pressure conditions used in the reaction. more, about 55% or more, about 75% or more, or about 90% Useful solvents are not particularly restricted provided that O. O. they do not interfere with the reaction itself (that is, they It is to be understood that wherever values and ranges are 55 preferably are inert solvents), and they dissolve a certain provided herein, e.g., in ages of subject populations, dosages, amount of the reactants. Depending on the circumstances, and blood levels, all values and ranges encompassed by these solvents may be distilled or degassed. Solvents may be, for values and ranges, are meant to be encompassed within the example, aliphatic hydrocarbons (e.g., hexanes, heptanes, scope of the present invention. Moreover, all values in these ligroin, petroleum ether, cyclohexane, or methylcyclohex values and ranges may also be the upper or lower limits of a 60 ane) and halogenated hydrocarbons (e.g., methylenechloride, range. chloroform, carbontetrachloride, dichloroethane, chloroben Furthermore, the invention pertains to any novel chemical Zene, or dichlororbenzene); aromatic hydrocarbons (e.g., compound described herein. That is, the invention relates to benzene, toluene, tetrahydronaphthalene, ethylbenzene, or novel compounds, and novel methods of their use as Xylene); ethers (e.g., diglyme, methyl-tert-butyl ether, described herein, which are within the scope of the Formulae 65 methyl-tert-amyl ether, ethyl-tert-butyl ether, diethylether, disclosed herein, and which are not disclosed in the cited diisopropylether, tetrahydrofuran or methyltetrahydrofurans, Patents and patent applications. dioxane, dimethoxyethane, or diethyleneglycol dimethyl US 8,642,801 B2 187 188 ether); nitriles (e.g., acetonitrile); ketones (e.g., acetone); lyophilized). In another aspect of the invention, the agents esters (e.g., methyl acetate or ethyl acetate); and mixtures and buffers necessary for carrying out the methods of the thereof. invention may be packaged as a kit. The kit may be commer In general, after completion of the reaction, the product is cially used according to the methods described herein and isolated from the reaction mixture according to standard tech may include instructions for use in a method of the invention. niques. For example, the solvent is removed by evaporation or Additional kit components may include acids, bases, buffer filtration if the product is solid, optionally under reduced ing agents, inorganic salts, solvents, antioxidants, preserva pressure. After the completion of the reaction, water may be tives, or metal chelators. The additional kit components are added to the residue to make the aqueous layer acidic or basic present as pure compositions, or as aqueous or organic solu and the precipitated compound filtered, although care should 10 tions that incorporate one or more additional kit components. be exercised when handling water-sensitive compounds. Any or all of the kit components optionally further comprise Similarly, water may be added to the reaction mixture with a buffers. hydrophobic solvent to extract the target compound. The The therapeutic agent may also be administered parenter organic layer may be washed with water, dried over anhy ally, intraperitoneally, intraspinally, or intracerebrally. Dis drous magnesium Sulphate or Sodium Sulphate, and the Sol 15 persions can be prepared in glycerol, liquid polyethylene vent is evaporated to obtain the target compound. The target glycols, and mixtures thereof and in oils. Under ordinary compound thus obtained may be purified, if necessary, e.g., conditions of storage and use, these preparations may contain by recrystallization, reprecipitation, chromatography, or by a preservative to prevent the growth of microorganisms. converting it to a salt by addition of an acid or base. To administer the therapeutic agent by other than The compounds of the invention may be supplied in a parenteral administration, it may be necessary to coat the Solution with an appropriate solvent or in a solvent-free form agent with, or co-administer the agent with, a material to (e.g., lyophilized). In another aspect of the invention, the prevent its inactivation. For example, the therapeutic agent compounds and buffers necessary for carrying out the meth may be administered to a Subject in an appropriate carrier, for ods of the invention may be packaged as a kit, optionally example, liposomes, or a diluent. Pharmaceutically accept including a container. The kit may be commercially used for 25 able diluents include Saline and aqueous buffer Solutions. treating or preventing amyloid-related disease according to Liposomes include water-in-oil-in-water CGF emulsions as the methods described herein and may include instructions well as conventional liposomes (Strean et al., J. Neuroimmu for use in a method of the invention. Additional kit compo mol. 7, 27 (1984)). nents may include acids, bases, buffering agents, inorganic Pharmaceutical compositions suitable for injectable use salts, solvents, antioxidants, preservatives, or metal chelators. 30 include sterile aqueous solutions (where water soluble) or The additional kit components are present as pure composi dispersions and sterile powders for the extemporaneous tions, or as acqueous or organic solutions that incorporate one preparation of sterile injectable solutions or dispersion. In all or more additional kit components. Any or all of the kit cases, the composition must be sterile and must be fluid to the components optionally further comprise buffers. extent that easy Syringability exists. It must be stable under The term “container includes any receptacle for holding 35 the conditions of manufacture and storage and must be pre the therapeutic compound. For example, in one embodiment, served against the contaminating action of microorganisms the container is the packaging that contains the compound. In Such as bacteria and fungi. other embodiments, the container is not the packaging that Suitable pharmaceutically acceptable vehicles include, contains the compound, i.e., the container is a receptacle, without limitation, any non-immunogenic pharmaceutical Such as a box or vial that contains the packaged compound or 40 adjuvants Suitable for oral, parenteral, nasal, mucosal, trans unpackaged compound and the instructions for use of the dermal, intravascular (IV), intraarterial (IA), intramuscular compound. Moreover, packaging techniques are well known (IM), and Subcutaneous (SC) administration routes. Such as in the art. It should be understood that the instructions for use phosphate buffer saline (PBS). of the therapeutic compound may be contained on the pack The vehicle can be a solvent or dispersion medium con aging containing the therapeutic compound, and as such the 45 taining, for example, water, ethanol, polyol (for example, instructions form an increased functional relationship to the glycerol, propylene glycol, and liquid polyethylene glycol, packaged product. and the like), suitable mixtures thereof, and vegetable oils. Pharmaceutical Preparations The proper fluidity can be maintained, for example, by the use In another embodiment, the present invention relates to of a coating Such as lecithin, by the maintenance of the pharmaceutical compositions comprising agents according to 50 required particle size in the case of dispersion and by the use any of the Formulae herein for the treatment of an amyloid of Surfactants. Prevention of the action of microorganisms related disease, as well as methods of manufacturing Such can be achieved by various antibacterial and antifungal pharmaceutical compositions. agents, for example, parabens, chlorobutanol, phenol, ascor In general, the agents of the present invention may be bic acid, thimerosal, and the like. In many cases, isotonic prepared by the methods illustrated in the general reaction 55 agents are included, for example, Sugars, sodium chloride, or schemes as, for example, in the patents and patent applica polyalcohols such as mannitol and Sorbitol, in the composi tions referred to herein, or by modifications thereof, using tion. Prolonged absorption of the injectable compositions can readily available starting materials, reagents and conven be brought about by including in the composition an agent tional synthesis procedures. In these reactions, it is also pos which delays absorption, for example, aluminum monostear sible to make use of variants which are in themselves known, 60 ate or gelatin. but are not mentioned here. Functional and structural equiva Sterile injectable solutions can be prepared by incorporat lents of the agents described herein and which have the same ing the therapeutic agent in the required amount in an appro general properties, wherein one or more simple variations of priate solvent with one or a combination of ingredients enu substituents are made which do not adversely affect the essen merated above, as required, followed by filtered sterilization. tial nature or the utility of the agent are also included. 65 Generally, dispersions are prepared by incorporating the The agents of the invention may be Supplied in a solution therapeutic agent into a sterile vehicle which contains a basic with an appropriate solvent or in a solvent-free form (e.g., dispersion medium and the required other ingredients from