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`ANNALS OF THE NEW YORK ACADEMY OF SCIENCES
`Volume 640
`
`AGING AND ALZHEIMER'S
`DISEASE
`
`SENSORY SYSTEMS, ~EURONAL GROWTH,
`AND NEURONAL METABOLISM
`
`Edited by John H. Growdon, Suzanne Corkin, Eva Ritter- Walker,
`and Richard J. Wurtman
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`IPR2014-00550
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`Pharmacologic and
`Clinicopharmacologic Properties of
`SDZ ENA 713, a Centrally Selective
`Acetylcholinesterase Inhibitor
`
`ALBERT ENZ AND HENDRIKUS BODDEKE
`Preclinical Research CNS Department
`Sandoz Pharma Ltd.
`CH-4002 Basle, Switzerland
`JULIAN GRAY AND RENE SPIEGEL
`Clinical Research CNS Department
`Sandoz Pharma Ltd.
`CH-4002 Basle, Switzerland
`
`. 1
`
`Among several clinically tested approaches to increase brain cholinergic activity, the
`acetylcholinesterase inhibitors (AChEIs) seem to represent the most promising sub(cid:173)
`stance class for treating Alzheimer's disease (AD). However, most compounds used so
`far in clinical trials lack brain selectivity, and their administration is accompanied by
`strong peripheral cholinergic or toxic effects. A cholinergic agent with an ideal profile
`should stimulate, directly or indirectly, central cholinergic receptors with an adequate
`duration of action and without influencing peripheral, particularly cardiovascular,
`systems. Physostigmine, a frequently used and very potent AChEI, has two serious
`disadvantages: its short biologic half-life and its unpredictable bioavailability after oral
`administration (Levy et al. 1986; Becker and Giacobini 1988).
`We describe here some pharmacologic properties of a novel AChEI, SDZ ENA
`713, that readily penetrates the central nervous system after parenteral and oral admin(cid:173)
`istration. This drug appears to have greater chemical stability and longer duration of
`action than does physostigmine. We also present first clinicopharmacologic data ob(cid:173)
`tained with SDZ ENA 713 in healthy volunteers.
`
`PHARMACOLOGIC METHODS
`
`Activity of AChE was determined according to the method described by Ellman
`et al. (1961). Rat brain tissue was homogenized in cold 0.25 mM phosphate buffer, pH
`7.4, containing 1 % Triton X-l00. After centrifugation, aliquots of the clear supernatant
`were used as enzyme source. AChE activity in different rat brain regions ex vivo was
`measured in similar extracts in which animals received various single doses of SDZ
`ENA 713 or physostigmine orally and subcutaneously, respectively, 30 minutes before
`sacrifice by decapitation.
`Hippocampal electroencephalographic activity from anesthetized male Sprague(cid:173)
`Dawley rats was recorded as described by Bevan (1984). The recording electrodes were
`implanted at the following coordinates (from bregma): A: -0.4 mm, L: 2.0 mm; and
`V: -2.5 mm.
`
`272
`
`NOVARTIS EXHIBIT 2026
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`IPR2014-00550
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`ENZ et al.: SDZ ENA 713
`
`273
`
`100.-r----------r----------r----------r------~
`
`% AChE
`Inhibit ion
`+/-SEM, n=6
`
`90
`
`80
`
`70
`
`60
`
`50
`
`40
`
`30
`
`20
`
`10
`
`" .... ·0· ...... Cortex, Physostigmine s.c.
`.. ...... A; .. - Hippocampus, Physost Igmlne s. C .
`.. _ .. 0-...... Striatum, Physostigmi ne s.c .
`
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`
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`....... <> ....... ponS/Medul .la, Physostigmine S'C' i/·
`
`................ Heart, Physostigmine s.c.
`-
`Cortex, ENA 713 S. c.
`- j . - Hippocampus, ENA 713 s.c.
`-
`Striatum, ENA 713 s.c.
`--+- Pons/Medulla, ENA 713 S.c.
`-x - Heart, ENA 713 s.c .
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`1.000
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`
`FIGURE 1. AChE inhibition by SDZ ENA 713 and physostigmine in rat brain and heart.
`Horizontal axis: Dose (p.mollkg) s.c. 30 min before sacrificing.
`
`PHARMACOLOGIC RESULTS
`
`ENA 713 inhibited AChE in a time-dependent manner. The compounds was found
`to be 100 times less potent than was physostigmine (Ki values ENA 713: 1-2 X 10-6
`M; physostigmine: 1-2 X 10- 8 M) in vitro, but the difference was reduced to a factor
`of 10 in in vivo experiments (FIG. 1). SDZ ENA 713 inhibited AChE in rat hippocampus
`and cortex (two brain regions primarily affected in AD) more potently than in other
`regions such as striatum and pons/medulla or than in the heart. This selectivity
`was not observed with physostigmine. A characteristic property of central muscarinic
`stimulation by either muscarinic agonists or AChEI is the induction of slow rhythmic
`activity (synchronization of theta waves) in the rat hippocampus (Bevan 1984). Intra(cid:173)
`peritoneal administration ofSDZ ENA 713 to anesthetized rats induced a pronounced
`increase in the hippocampal theta rhythm (+ 120% with 0.8 mglkg), a similar effect
`being obtained with physostigmine. Following oral administration of SDZ ENA 713,
`the effect was also pronounced (+ 170%). Up to a dose of 1.5 mg/kg intravenously (a
`dose inducing marked central cholinergic activation), SDZ ENA 713 showed no effects
`on circulatory parameters in the anesthetized cat (Enz et al. 1989).
`
`FINDINGS IN HEALTHY HUMAN VOLUNTEERS
`
`Eighty young male subjects took part in a study of the tolerability and pharmacody(cid:173)
`namic effects of single rising oral doses of SDZ ENA 713. Eight groups of 10 subjects
`(7 on active medication, 3 on placebo in each group) received single rising oral doses
`
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`NOVARTIS EXHIBIT 2026
`Noven v. Novartis and LTS Lohmann
`IPR2014-00550
`Page 3 of 5
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`

`274
`
`ANNALS NEW YORK ACADEMY OF SCIENCES
`
`BChE act i v ity
`% predrug
`+/- SEM
`
`llO
`
`105
`
`100
`
`95
`
`90
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`85
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`75
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`70
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`65
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`60
`
`~ Placebo
`
`-.- Group 5: 1.5 mg
`
`.......•....... Group 6: 2 . 3 mg
`- - A - - Group 7: 3 mg
`-,- ,0-, - Group 8: 4.6 mg
`
`4
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`6
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`10
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`12
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`14
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`16
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`18
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`20
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`
`24
`
`Time after drug administration (hours)
`FIGURE 2. BChE activities in human plasma after single oral ENA 713 administration.
`
`of 0.16, 0.30, 0.60, 1.0, 1.5,2.3,3.0, and 4.6 mg ofSDZ ENA 713 or placebo. Tolerabil(cid:173)
`ity up to the 3-mg dose was generally good. After the 4.6-mg dose, nausea, vomiting,
`headache, or dizziness was observed in four of seven subjects after active medication.
`However, peripheral cholinergic signs such as sweating and salivation were absent or
`minimal. Vital signs and routine hematologic and biochemical parameters were not
`significantly altered at any dose level.
`Dose-dependent inhibition of plasma butyrylcholinesterase was observed with a
`duration of action of greater than 10 hours (FIG. 2). In a separate study, single doses
`of 1.3 and 2.0 mg were centrally active, inducing an increase in the density of rapid
`eye movements during REM sleep in healthy young volunteers. Therefore, the concept
`of SDZ ENA 713 as a centrally selective, long-acting acetylcholinesterase inhibitor is
`so far confirmed by experience in man. Subsequent experience with single and multiple
`dose administration in young and elderly volunteers and patients with AD have contin(cid:173)
`ued to support this concept. The compound is presently being tested as a symptomatic
`treatment for AD.
`
`REFERENCES
`
`BECKER, R. E. & E. GIACOBINI. 1988. Mechanisms of cholinesterase inhibition in senile dementia
`of the Alzheimer type: Clinical, pharmacological and therapeutic aspects. Drug Dev. Res. 12:
`163-195.
`.
`BEVAN, P. 1984. Effect of muscarinic ligands on the electrical activity recorded from the hippo(cid:173)
`campus: A quantitative approach. Br. J. Pharmacol. 82: 431-440.
`ELLMAN, G . L., K. D. COURTNEY, V. ANDRES & R. M. FEATHERSTONE. 1961. A new rapid
`colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7: 88-95.
`ENZ, A., R. AMSTUTZ, A. HOFMANN, G. GMELIN & P. H. KELLY. 1989. Pharmacological
`properties of the preferentially centrally acting acetylcholinesterase inhibitor SDZ ENA 713.
`
`NOVARTIS EXHIBIT 2026
`Noven v. Novartis and LTS Lohmann
`IPR2014-00550
`Page 4 of 5
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`ENZ et al.: SDZ ENA 713
`
`275
`
`In Pharmacological Interventions on Central Cholinergic Mechanisms in Senile Dementia
`(Alzheimer's disease). H. Kewitz, T. Thomsen & M. Bickel, Eds.: 271-277. Zuckschwerdt
`Verlag. Munich.
`LEVY, D., P. GUKFELD, Y. GRUNFELD, J. GRUNWALD, M. KUSHNIR, A. LEVY, Y. MESHULAM,
`M. SPIEGELSTEIN, D. ZEHAVI & A. FISHER. 1986. A novel transdermal therapeutic system as
`a potential treatment for Alzheimer's disease. In Advances in Behavioral Biology, Vol. 29:
`557- 563. A. Fisher, I. Hanin & C. Lachman, Eds. Plenum Press. New York, NY .
`
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`Noven v. Novartis and LTS Lohmann
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