Abstract
Abnormal accumulation of neurotoxic β-amyloid peptides (Aβ) in brain represents a key factor in the progression of Alzheimer's disease (AD). Identification of small molecules that effectively reduce brain levels of Aβ is important for development of Aβ-lowering agents for AD. In this study, we demonstrate that in vivo Aβ levels in brain are significantly reduced by the cysteine protease inhibitor E64d and the related CA074Me inhibitor, which inhibits cathepsin B. Direct infusion of these inhibitors into brains of guinea pigs resulted in reduced levels of Aβ by 50–70% after 30 days of treatment. Substantial decreases in Aβ also occurred after only 7 days of inhibitor infusion, with a reduction in both Aβ40 and Aβ42 peptide forms. A prominent decrease in Aβ peptides was observed in brain synaptosomal nerve terminal preparations after CA074Me treatment. Analyses of APP-derived proteolytic fragments showed that CA074Me reduced brain levels of the CTFβ fragment, and increased amounts of the sAPPα fragment. These results suggest that CA074Me inhibits Aβ production by modulating APP processing. Animals appeared healthy after treatment with these inhibitors. These results, showing highly effective in vivo decreases in brain Aβ levels by these cysteine protease inhibitors, indicate the feasibility of using related compounds for lowering Aβ in AD.
References
Beck, M., Muller, D., and Bigl, V. (1997). Amyloid precursor protein in guinea pigs – complete cDNA sequence and alternative splicing. Biochim. Biophys. Acta1351, 17–21.10.1016/S0167-4781(96)00232-1Search in Google Scholar
Bernstein, H.G., Kirschke, H., Wiederanders, B., Schmidt, D., and Rinne, A. (1990). Antigenic expression of cathepsin B in aged human brain. Brain Res. Bull.24, 543–549.10.1016/0361-9230(90)90157-USearch in Google Scholar
Buttle, D.J., Murata, M., Knight, C.G., and Barrett, A.J. (1992). CA074 methyl ester: a proinhibitor for intracellular cathepsin B. Arch. Biochem. Biophys.299, 377–380.10.1016/0003-9861(92)90290-DSearch in Google Scholar
Cataldo, A.M. and Nixon, R.A. (1990). Enzymatically active lysosomal proteases are associated with amyloid deposits in Alzheimer brain. Proc. Natl. Acad. Sci. USA87, 3861–3865.10.1073/pnas.87.10.3861Search in Google Scholar PubMed PubMed Central
Cataldo, A.M., Paskevich, P.A., Kominami, E., and Nixon, R.A. (1991). Lysosomal hydrolases of different classes are abnormally distributed in brains of patients with Alzheimer's disease. Proc. Natl. Acad. Sci. USA88, 10998–11002.10.1073/pnas.88.24.10998Search in Google Scholar PubMed PubMed Central
Cirrito, J.R., Yamade, K.A., Finn, M.B., Sloviter, R.S., Bales, K.R., May, P.C., Schoepp, D., Paul, S.M., Mennerick, S., and Holtzman, D.M. (2005). Synaptic activity regulates interstitial fluid amyloid-β levels in vivo. Neuron48, 913–922.10.1016/j.neuron.2005.10.028Search in Google Scholar PubMed
Citron, M., Oltersdorf, T., Haass, C., McConlogue, L., Hung, A.Y., Seubert, P., Vigo-Pelfrey, C., Lieberburg, I., and Selkoe, D.J. (1992). Mutation of the β-amyloid precursor protein in familial Alzheimer's disease increases β-protein production. Nature360, 672–674.10.1038/360672a0Search in Google Scholar PubMed
Deane, R., Du Yan, S., Submamaryan, R.K., Larue, B., Jovanovic, S., Hogg, E., Welch, D., Manness, L., Lin, C., Yu, J., et al. (2003). RAGE mediates amyloid-β peptide transport across the blood-brain barrier and accumulation in brain. Nat. Med.9, 907–913.10.1038/nm890Search in Google Scholar PubMed
Farber, S.A., Nitsch, R.M., Schulz, J.G., and Wurtman, R.J. (1995). Regulated secretion of beta-amyloid precursor protein in rat brain. J. Neurosci.15, 7442–7451.10.1523/JNEUROSCI.15-11-07442.1995Search in Google Scholar
Hook, V.Y.H., Toneff, T., Aaron, W., Yasothornsrikul, S., Bundey, R., and Reisine, T. (2002). β-Amyloid peptide in regulated secretory vesicles of chromaffin cells: evidence for multiple cysteine proteolytic activities in distinct pathways for β-secretase activity in chromaffin vesicles. J. Neurochem.81, 237–256.10.1046/j.1471-4159.2002.00794.xSearch in Google Scholar PubMed
Hook, V., Toneff, T., Bogyo, M., Greenbaum, D., Medzihradszky, K.F., Neveu, J., Lane, W., Hook, G., and Reisine, T. (2005). Inhibition of cathepsin B reduces β-amyloid production in regulated secretory vesicles of neuronal chromaffin cells: evidence for cathepsin B as a candidate β-secretase of Alzheimer's disease. Biol. Chem.386, 931–940.Search in Google Scholar
Iversen, L.L., Mortishire-Smith, R.J., Pollack, S.J., and Shearman, M.S. (1995). The toxicity in vitro of β-amyloid protein. Biochem. J.311, 1–16.10.1042/bj3110001Search in Google Scholar
Ivy, G.O., Schottler, F., Wenzel, J., Baudry, M., and Lynch, G. (1984). Inhibitors of lysosomal enzymes: accumulation of lipofuscin-like dense bodies in the brain. Science226, 985–987.10.1126/science.6505679Search in Google Scholar
Johnstone, E.M., Chaney, M.O., Norris, F.H., Pascual, R., and Little, S.P. (1991). Conservation of the sequence of the Alzheimer's disease amyloid peptide in dog, polar bear and five other mammals by cross-species polymerase chain reaction analysis. Brain Res.10, 299–305.10.1016/0169-328X(91)90088-FSearch in Google Scholar
Komatsu, K., Inazuki, K., Hosoya, J., and Satoh, S. (1986). Beneficial effect of new thiol protease inhibitors, epoxide derivatives, on dystrophic mice. Exp. Neurol.91, 23–29.10.1016/0014-4886(86)90022-1Search in Google Scholar
Lin, S., Koelsch, G., Wu, S., Downs, D., Dashti, A., and Tang, J. (2000). Human aspartic protease memapsin 2 cleaves the β-secretase site of β-amyloid precursor protein. Proc. Natl. Acad. Sci. USA97, 1456–1460.10.1073/pnas.97.4.1456Search in Google Scholar
Marr, R.A., Guan, H., Rockenstien, E., Kindy, M., Gage, F.H., Verma, I., Masliah, E., and Hersh, L.B. (2004). Neprilysin regulates amyloid beta peptide levels. J. Mol. Neurosci.22, 5–11.10.1385/JMN:22:1-2:5Search in Google Scholar
Miyahara, T., Shimojo, S., Toyohara, K., Imai, T., Miyajima, M., Honda, H., Kamegai, M., Ohzeki, M., and Kokatsu, J. (1985). Phase I study of EST, a new thiol protease inhibitor – 2nd report: safety and pharmacokinetics in continuous administration. Rinsho Yakuri16, 537–546.10.3999/jscpt.16.537Search in Google Scholar
Mueller-Steiner, S., Zhou, Y., Arai, H., Roberson, E.D., Sun, B., Chen, J., Wang, X., Yu, G., Esposito, L., Mucke, L., and Gan, L. (2006). Antiamyloidogenic and neuroprotective functions of cathepsin B: implications for Alzheimer's disease. Neuron51, 703–714.10.1016/j.neuron.2006.07.027Search in Google Scholar
Murray, E.J., Griasnti, J.S., Bentley, G.V., and Murray, S.S. (1997). E64d, a membrane-permeable cysteine protease inhibitor, attenuates the effects of parathyroid hormone on osteoblasts in vitro. Metabolism46, 1090–1094.10.1016/S0026-0495(97)90284-5Search in Google Scholar
Nakamura, Y., Takeda, M., Suzuki, H., Hattori, H., Tada, K., Hariguchi, S., Hashimoto, S., and Nishimura, T. (1991). Abnormal distribution of cathepsins in the brain of patients with Alzheimer's disease. Neurosci. Lett.130, 195–198.10.1016/0304-3940(91)90395-ASearch in Google Scholar
Nakanishi, H., Tominaga, K., Amano, T., Hirotsu, I., Inoue, T., and Yamamoto, K. (1994). Age-related changes in activities and localizations of cathepsins D, E, B, and L in the rat brain tissues. Exp. Neurol.126, 119–128.10.1006/exnr.1994.1048Search in Google Scholar
Nitsch, R.M., Farber, S.A., Growdon, J.H., and Wurtman, R.J. (1993). Release of amyloid beta-protein precursor derivatives by electrical depolarization of rat hippocampal slices. Proc. Natl. Acad. Sci. USA90, 5191–5193.10.1073/pnas.90.11.5191Search in Google Scholar
Satoyashi, E. (1992). Therapeutic trials on progressive muscular dystrophy. Intern. Med.31, 841–846.10.2169/internalmedicine.31.841Search in Google Scholar
Selkoe, D.J. (2001). Alzheimer's disease: genes, proteins, and therapy. Physiol. Rev.81, 741–766.10.1152/physrev.2001.81.2.741Search in Google Scholar
Sinha, S., Anderson, J.P., Barbour, R., Basi, G.S., Caccavello, R., Davis, D, Doan, M., Dovey, F.H., Frigon, N., Hong, J., et al. (1999). Purification and cloning of amyloid precursor protein β-secretase from human brain. Nature402, 537–540.10.1038/990114Search in Google Scholar
Sisodia, S.S. (1999). Alzheimer's disease: perspectives for the new millennium. J. Clin. Invest.104, 1169–1170.10.1172/JCI8508Search in Google Scholar
Tamai, M., Matsumoto, K., Omura, S., Koyama, I., Ozawa, Y., and Hanada, K. (1986). In vitro and in vivo inhibition of cysteine proteinases by EST, a new analog of E-64. J. Pharmacobiodyn.9, 672–677.10.1248/bpb1978.9.672Search in Google Scholar
Tamai, M., Omura, S., Kimura, M., Hanada, K., and Sugita, H. (1987). Prolongation of life span of dystrophic hamster by cysteine proteinase inhibitor, loxistatin (EST). J. Pharmacobiodyn.10, 678–681.10.1248/bpb1978.10.678Search in Google Scholar
Towatari, T., Nikawa, T., Murata, M., Yokoo, C., Tamai, M., Hanada, K., and Katunuma, N. (1991). Novel epoxysuccinyl peptides, a selective inhibitor of cathepsin B in vivo. FEBS Lett.280, 311–315.10.1016/0014-5793(91)80319-XSearch in Google Scholar
Tsuchiya, K., Kohda, Y., Yoshikda, M., Zhao, L., Ueono, T., Yamashita, J., Yoshioka, T., Kominami, E., and Yamashima, T. (1999). Postictal blockade of ischemic hippocampal neuronal death in primates using selective cathepsin inhibitors. Exp. Neurol.155, 187–194.10.1006/exnr.1998.6988Search in Google Scholar PubMed
Vassar, R., Bennet, B.D., Babu-Khan, S., Mendiaz, E.A., Denis, P., Teplow, D.B., Ross, S., Amarante, P., Loeloff, R., Luo, Y., et al. (1999). β-Secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science286, 735–741.10.1126/science.286.5440.735Search in Google Scholar
Verhage, M., Ghijsen, W.E., Nicholls, D.G., and Wiegant, V.M. (1991a). Characterization of the release of cholecystokinin-8 from isolated nerve terminals and comparisons with exocytosis of classical transmitters. J. Neurochem.56, 1394–1400.10.1111/j.1471-4159.1991.tb11437.xSearch in Google Scholar
Verhage, M., McMahon, H.T., Ghijsen, W.E., Boomsma, F., Scholten, G., Wiegant, V.M., and Nichols, D.G. (1991b). Differential release of amino acids, neuropeptides, and catecholamines from isolated nerve terminals. Neuron6, 517–524.10.1016/0896-6273(91)90054-4Search in Google Scholar
Yan, R., Bienkowski, M.J., Shuck, M.E., Miao, H., Tory, M.C., Pauley, A.M., Brashier, J.R., Stratment, N.C., Mathews, W.R., Buhl, A.E., et al. (1999). Membrane-anchored aspartyl protease with Alzheimer's disease β-secretase activity. Nature402, 533–537.10.1038/990107Search in Google Scholar PubMed
Yoshida, M., Yamashima, T., Zhao, L., Tsuchiya, K., Kohda, Y., Tonchev, A.B., Matsuda, M., and Kominami, E. (2002). Primate neurons show different vulnerability to transient ischemia and response to cathepsin inhibition. Acta Neuropathol.104, 267–272.10.1007/s00401-002-0554-4Search in Google Scholar PubMed
Zhang, J., Goodlett, D.R., Quinn, J.F., Peskind, E., Kaye, J.A., Zhou, Y., Pan, C., Yi, E., Eng, J., Wang, Q., et al. (2005) Quantitative proteomics of cerebrospinal fluid from patients with Alzheimer's disease. J. Alzheimer's Dis.7, 125–133.10.3233/JAD-2005-7205Search in Google Scholar PubMed
©2007 by Walter de Gruyter Berlin New York
