U.S. patent number RE39,616 [Application Number 11/091,008] was granted by the patent office on 2007-05-08 for aminoindan derivatives.
This patent grant is currently assigned to Technion Research and Development Foundation, Ltd., Teva Pharmaceutical Industries, Ltd., Yissum Research Development Company of the Hebrew University of Jerusalem. Invention is credited to Michael Chorev, Tamar Goren, Yacov Herzig, Jeffrey Sterling, Marta Weinstock-Rosin, Moussa B. H. Youdim.
United States Patent |
RE39,616 |
Chorev , et al. |
May 8, 2007 |
Aminoindan derivatives
Abstract
This invention is directed to compounds of the following
formula: ##STR00001## wherein when a is 0, b is 1 or 2; when a is
1, b is 1, m is from 0-3, X is O or S, Y is halogeno, R.sub.1 is
hydrogen C.sub.1-4 alkyl, R.sub.2 is hydrogen, C.sub.1-4 alkyl, or
optionally substituted propargyl and R.sub.3 and R.sub.4 are each
independently hydrogen, C.sub.1-6 alkyl, C.sub.6-12 aryl,
C.sub.6-12 aralkyl each optionally substituted. This invention is
also directed to the use of these compounds for treating
depression, Attention Deficit Disorder (ADD), Attention Deficit and
Hyperactivity Disorder (ADHD), Tourette's Syndrome, Alzheimer's
Disease and other dementia's such as senile dementia, dementia of
the Parkinson's type, vascular dementia and Lewy body dementia.
This invention is further directed to a pharmaceutical composition
comprising a therapeutically effective amount of the above-defined
compounds and a pharmaceutically acceptable carrier.
Inventors: |
Chorev; Michael (Jerusalem,
IL), Goren; Tamar (Rehovot, IL), Herzig;
Yacov (Ra'ananna, IL), Sterling; Jeffrey
(Jerusalem, IL), Weinstock-Rosin; Marta (Jerusalem,
IL), Youdim; Moussa B. H. (Haifa, IL) |
Assignee: |
Teva Pharmaceutical Industries,
Ltd. (Petach-Tikva, IL)
Yissum Research Development Company of the Hebrew University of
Jerusalem (Jerusalem, IL)
Technion Research and Development Foundation, Ltd. (Haifa,
IL)
|
Family
ID: |
26323338 |
Appl.
No.: |
11/091,008 |
Filed: |
March 25, 2005 |
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
09336493 |
Jun 18, 1999 |
|
|
|
|
PCT/US97/24155 |
Dec 18, 1997 |
|
|
|
Reissue of: |
09944912 |
Aug 31, 2001 |
06538025 |
Mar 25, 2003 |
|
|
Foreign Application Priority Data
|
|
|
|
|
Dec 18, 1996 [IL] |
|
|
119853 |
Mar 24, 1997 [IL] |
|
|
120510 |
|
Current U.S.
Class: |
514/480; 560/115;
560/29; 560/32; 560/136; 514/481 |
Current CPC
Class: |
A61P
25/00 (20180101); C07C 333/04 (20130101); C07C
271/56 (20130101); C07C 271/44 (20130101); A61P
25/28 (20180101); C07C 271/58 (20130101); C07C
271/24 (20130101); C07C 2601/14 (20170501); C07C
2602/10 (20170501); C07C 2602/08 (20170501) |
Current International
Class: |
A61K
31/27 (20060101) |
Field of
Search: |
;514/480,481
;560/29,32,115,136 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
|
|
|
|
|
0436492 |
|
Oct 1991 |
|
EP |
|
0664291 |
|
Oct 1993 |
|
EP |
|
0614888 |
|
Feb 1994 |
|
EP |
|
0538134 |
|
Mar 1997 |
|
EP |
|
0852735 |
|
Nov 1960 |
|
GB |
|
1003686 |
|
Sep 1965 |
|
GB |
|
3002155 |
|
Mar 1991 |
|
JP |
|
WO9100724 |
|
Jan 1991 |
|
WO |
|
WO9504027 |
|
Feb 1995 |
|
WO |
|
WO9511016 |
|
Apr 1995 |
|
WO |
|
WO9518617 |
|
Jul 1995 |
|
WO |
|
WO9602524 |
|
Feb 1996 |
|
WO |
|
Other References
Armstrong et al., "Acylation Effects on Chiral Recognition of
Racemic Amines and Alcohols by New Polar and Non-Polar Cyclodextrin
Derivative Gas Chromatographic Phases", J. Chromatography (1990)
502: 154-159. cited by other .
Askin et al., "Highly Diastereoselective Alkylations of Chiral
Amide Enolates: New Routes to Hydroxyethylene Dipeptide Isostere
Inhibitors of HIV-1 Protease", J. Org. Chem. (1992) 57(10):
2771-2773. cited by other .
Baker et al., "Synthesis of Decahydrocyclopentacyclo-octene
Derivatives via Intramolecular Photocycloaddition of Butenolides
and Reductive Cleavage", J. Chem. Soc. Chem. Comm. (1980) 23:
1011-1012. cited by other .
Barton et al., "Reductive Formylation of Oximes; An Approach to the
Synthesis of Vinyl Isonitriles", Tetrahedron Letters (1988) 29(27):
3343-3346. cited by other .
Boar et al., "A Simple Synthesis of Enamides from Ketoximes", J.
Chem. Soc. Perkins I (1975) 1237-1241. cited by other .
Brettle et al., "Synthesis of Enamides", J. Chem. Soc. Perkin
Trans. I, (1988) 2185-2193. cited by other .
Burk et al., "A Three-Step Procedure for Asymmetric Catalytic
Reductive Amidation of Ketones", J. Org. Chem. (1998) 63:
6084-6085. cited by other .
Chorvat el al., "Acetycholine Release Enhancing Agents: Potential
Therapeutics For Alzheimer's Disease", Drugs of the Future (1995)
20(11):1145-1162. cited by other .
Cooper et al., "Alzheimer's Disease Drug Treatment", J. Ger. Drug
Ther., (1993) 8(2):5-18. cited by other .
Cutler et al., "Muscarinic M1-Receptor Agonists", CNS Drugs (1995)
3(6):467-481. cited by other .
Davis et al., "Tacrine", The Lancet (1995) 345:625-630. cited by
other .
Drefahl et al., "Amino Alcohols. I. Cis- and
Trans-DL-1-amino-2-hydroxytetrahydronaphthalene and Cis- and
Trans-DL-1-amino-2-hydroxyindan", Chem. Abstracts (1958) 52:
16417f. cited by other .
Drefahl et al., "Amino Alcohols. X. Addition of Iodine Isocyanate
to Unsymmetrical Olefins", Chem. Abstracts (1960) 54: 13078f. cited
by other .
Finberg and Youdim, "Modification of Blood Pressure and Nictitating
Membrane Response to Sympathetic Amines by Selective Monoamine
Oxidase Inhibitors, Types A and B, in the Cat", Brit. J. Pharmacol.
(1985) 85(2): 541-546. cited by other .
Fink et al., "Imino 1,2,3,4-Tetrahydrocyclopent[B]indole Carbamates
as Dual Inhibitors of Acetylcholinesterase and Monoamine Oxidase",
Bioorganic & Medicinal Chemistry Letters (1996) 6(6): 625-630.
cited by other .
Fuller et al., "Inhibition in vitro of Norepinephrine
N-methyltransferase by 2-Aminotetralins, Analogs of
Phenylethylamines with Rigid Conformation", Biochem. Pharmacol.,
(1976) 26: 446-447. cited by other .
Gabryel et al., "Nootropics: Pharmacological Properties and
Therapeutic Use", Pol. J. Pharmacol. (1994) 46:383-394. cited by
other .
Ghislandi et al., "Scissione Ottica E Configurazione
Dell'1-Aminobenzociclobutene E Dell'1-Aminoindano", Boll. Chim.
Farm. (1976) 115: 489-500. cited by other .
Harvey, "The Pharmacology of Galanthamine and its Analogues",
Pharmac. Ther. (1995) 68(1):113-128. cited by other .
Heikkila et al., "Prevention of MPTP-Induced Neurotoxicity by
AGN-1133 and AGN-1135, Selective Inhibitors of Monoamine
Oxidase-B", Eur. J. Pharmacol. (1985) 116: 313-317. cited by other
.
Hori et al., "N-containing Diphenylethylamine Derivatives and Acid
Adducts", Japan Kokai Tokyo Koho JP 54-132559, Oct. 15, 1979,
Database CAPLUS on STN.RTM., Chemical Abstracts Service, (Columbus,
Ohio), Accession No. 1980:180807, abstract. cited by other .
Horn et al., "Steric Requirements for Catecholamine Uptake by Rat
Brain Synaptosomes: Studies with Rigid Analogs of Amphetamine", J.
Pharmacol. Exp. Ther. (1972) 180: 523-530. cited by other .
Huebner, "1-(N-Methyl-N-propargylamino)indans and Related
Compounds", Chem. Abstracts (1964) 61:3046a. cited by other .
Kabins et al., "Potential Applications for Monoamine Oxidase B
Inhibitors", Dementia (1990) 1: 323-348. cited by other .
Kametani et al., "Studies on the Syntheses of Heterocyclic
Compounds. CLIX. The Reaction of 2-Nitro-1-indanone Oxime with
Formalin and Hydrochloric Acid", Chem. Pharm. Bull. (1966) 14(12):
1408-1413. cited by other .
Knapp et al., "A 30-Week Randomized Controlled Trial of High-Dose
Tacrine in Patients with Alzheimer's Disease", J.A.M.A. (1994)
271(13) 985-991. cited by other .
Laso et al., "A New Selective Reduction of Nitroalkenes into
Enamides", Tetrahedron Letters (1996) 37(10): 1605-1608. cited by
other .
Martin et al., "Potential Anti-Parkinson Drugs Designed by Receptor
Mapping", J. Med. Chem. (1973) 16(2): 147-150. cited by other .
Martin et al., "Discriminant Analysis of the Relationship Between
Physical Properties and the Inhibition of Monoamine Oxidase by
Aminotetralins and Aminoindans", J. Med. Chem. (1974) 17(4):
409-413. cited by other .
Mouna et al., "Enantioselective Acetylation of Primary Amines by
Cylindrocarpon Radicicola", Bioorg. & Med. Chem. Letters.
(1993) 3(4): 681-684. cited by other .
Nakanishi et al., "Preparation of Enamides via Reductive Acylation
of N-Acetoxyimino Compounds by Use of Fe.sub.3 (CO).sub.12"
Chemistry Letters (1987) 2167-2168. cited by other .
O'Malley et al., "Synthesis and Biological Evaluation of Combined
Acetyl Cholinesterase (AChE) and Monoamine Oxidase (MAO)
Inhibitors", 205.sup.th ACS Mtg., 1993 (MEDI), abstract 78. cited
by other .
Oshiro et al., "Novel Cerebroprotective Agents with Central Nervous
System Stimulating Activity. 1. Synthesis and Pharmacology of
1-Amino-7-hydroxyindan Derivatives", J. Med. Chem. (1991) 34(7):
2004-2013. cited by other .
Palermo et al., "Combined Acetylcholinesterase (AChE) and
Reversible Monoamine Oxidase (MAO) Inhibition as a Potential
Therapeutic Approach For Senile Dementia of the Alzheimer Type
(SDAT)", 205.sup.th ACS Mtg., 1993 (MEDI), abstract 77. cited by
other .
Riederer and Youdim, "Monoamine Oxidase Activity and Monoamine
Metabolism in Brains of Parkinson's Patients Treated with
l-Deprenyl", J. Neurochem. (1986) 46(5): 1359-1365. cited by other
.
Ruschig et al., "Preparation of 17-hydroxy-20-keto Steroids from
17(20)-en-20-acetamino Steroids", Chem. Ber. (1955) 88(6):883-894.
cited by other .
Singh et al., "Antimalarials. 7-Chloro-4-(substituted
amino)quinolines", J. Med. Chem. (1971) 14(4): 283-286. cited by
other .
Sramek et al., "Safety/Tolerability Trial of SDZ ENA 713 in
Patients with Probable Alzheimer's Disease", Life Sciences (1996)
58(15):1201-1207. cited by other .
Tariot et al. "Treatment of Alzheimer's Disease: Glimmers of Hope",
Chem. Ind. (1993) 20:801-3, 806-7. cited by other .
Tekes et al., "Effect of MAO Inhibitors on the Uptake and
Metabolism of Dopamine in Rat and Human Brain", Pol. J. Pharmacol.
Pharm. (1988) 40: 653-658. cited by other .
Teranishi et al., "Facile Synthesis of 6-Hydroxyindole and
6-Methoxyindole via Regioselective Friedel-Crafts Acylation and
Baeyer-Villiger Oxidation", Synthesis (1994) 1018-1020. cited by
other .
Terni et al., "Preparation of (aminoalkyl) phenyl
morpholinoalkylcarbamates and analogs as cholinesterase
inhibitors", WO 96/02524, Feb. 1, 1996, Database CAPLUS on
STN.RTM., Chemical Abstracts Service (Columbus, Ohio), Accession
No. 1996:340192, abstract. cited by other .
Top et al., "N-Alkylation of Nitriles with Tricarbonylchromium
Complexes of Benzyl and Related Alcohols as Synthetic
Intermediates. Further Development of the Ritter Reactions", J.C.S.
Chem. Comm. (1979) 224-225. cited by other .
Weinstock, "The Pharmacotherapy of Alzheimer's Disease Based on the
Cholinergic Hypothesis: an Update", Neurodegeneration (1995)
4:349-356. cited by other .
Youdim et al., "Monamine Oxidase" in Handbook of Experimental
Pharmacology, v. 90/I (Trendelenburg and Weiner, eds.,
Springer-Verlag, London: 1988) Chpt. 3, 119-192. cited by other
.
Zheng et al., "Asymmetric Synthesis of Amino Acid Derivatives via
an Electophilic Amination of Chiral Amide Cuprates with Li
t-Butyl-N-Tosyloxycarbamate", Tetrahedron Letters (1997) 38(16):
2817-2820. cited by other .
Zhu et al., "Asymmetric Rh-Catalyzed Hydrogenation of Enamides with
a Chiral 1,4-Bisphosphine Bearing Diphenylphosphino Groups", J.
Org. Chem. (1998) 63: 9590-9593. cited by other .
"Agent for Cognition Disorders Acetylcholinesterase Inhibitor",
Drugs of the Future, E-2020 (1991) 16(1): 16-18. cited by other
.
"Cognition Enhancer Acetylcholinesterase Inhibitor" Drugs of the
Future, E-2020 (1995) 20(1): 77-78. cited by other .
"Cognition Enhancer Acetylcholinesterase Inhibitor", Drugs of the
Future, TAK-147 (1995) 20(3): 248-250. cited by other .
The Merck Index (Windholz et al., eds., Merck & Co., Inc.,
Rahway, NJ, 10th ed., 1983) 149, 248-249. cited by other .
The Merck Manual of Diagnosis and Therapy, (Berkow et al., eds.,
Merck Sharp & Dohme Research Laboratories, 15.sup.th ed., 1987)
1030-1033. cited by other .
The Merck Manual of Diagnosis and Therapy, (Berkow et al., eds.,
Merck Sharp & Dohme Research Laboratories, 15.sup.th ed., 1987)
1054-1055. cited by other .
The Parkinson Study Group, "Effect of Deprenyl on the Progression
of Disability in Early Parkinson's Disease" New Eng. J. Med. (1989)
321(20: 1364-1371. cited by other .
The Parkinson Study Group, "Effects of Tocopherol and Deprenyl on
the Progression of Disability in Early Parkinson's Disease" New
Eng. J. Med. (1993) 328(3): 176-183. cited by other.
|
Primary Examiner: Barts; Samuel
Attorney, Agent or Firm: White, Esq.; John P. Cooper &
Dunham LLP
Parent Case Text
This application is a continuation of U.S. Ser. No. 09/336,493,
filed Jun. 18, 1999, a continuation of PCT International
Application No. PCT/US97/24155, filed Dec. 18, 1997, designating
the United States of America and claiming priority of Israeli
Patent Application Nos. 119853, filed Dec. 18, 1996 and 120510,
filed Mar. 24, 1997, the contents of which are hereby incorporated
by reference.
Claims
What is claimed is:
1. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of a compound having the structure: ##STR00019## wherein
when a is 0, b is 1 or 2.Iadd., and when a is 1, b is 1.Iaddend.;
.[.wherein when a is 1, b is 1,.]. m is 0-3.[.,.]. .Iadd.;
.Iaddend.X is O or S.[.,.]. .Iadd.; .Iaddend.Y is halogeno.[.,.].
.Iadd.; .Iaddend.R.sub.1 is hydrogen or C.sub.1-4 alkyl.[.,.].
.Iadd.; .Iaddend.R.sub.2 is hydrogen, C.sub.1-4 alkyl,
.Iadd.propargyl, .Iaddend.or .[.optionally.]. substituted
propargyl.Iadd.; .Iaddend.and R.sub.3 and R.sub.4 are each
independently .[.hydrogen,.]. C.sub.1-8 alkyl, C.sub.6-12 aryl,
C.sub.6-12 .[.aryl.]. .Iadd.aralkyl .Iaddend.or C.sub.6-12
cycloalkyl, each .Iadd.of which may be .Iaddend.optionally
substituted, .Iadd.or is hydrogen, such compound being a racemic
mixture, an enantiomer, or a salt thereof;.Iaddend. .[.a racemic
mixture, an enantiomer, or salt thereof,.]. .Iadd.so as .Iaddend.to
thereby treat the subject's depression.
2. The method of claim 1 wherein the enantiomer is the R
enantiomer.
3. The method of claim 1 wherein the enantiomer is the S
enantiomer.
4. The method of claim 1 wherein the compound has the structure:
##STR00020##
5. The method of claim 4 wherein the enantiomer is the R
enantiomer.
6. The method of claim 4 wherein the enantiomer is the S
enantiomer.
7. The method of claim 1 wherein the compound has the structure:
##STR00021##
8. The method of claim 7 wherein the enantiomer is the R
enantiomer.
9. The method of claim 7 wherein the enantiomer is the S
enantiomer.
10. A method of selectively inhibiting monoamine oxidase-B (MAO-B)
activity in the brain of a subject in need of such inhibition
comprising administering to the subject a therapeutically effective
amount of a compound having the structure: ##STR00022## wherein
when a is 0, b is 1 or 2.Iadd., and when a is 1, b is 1.Iaddend.;
.[.wherein when a is 1, b is 1,.]. m is 0-3.[.,.]. .Iadd.;
.Iaddend.X is O or S.[.,.]. .Iadd.; .Iaddend.Y is halogeno.[.,.].
.Iadd.; .Iaddend.R.sub.1 is hydrogen or C.sub.1-4 alkyl.[.,.].
.Iadd.; .Iaddend.R.sub.2 is hydrogen, C.sub.1-4 alkyl,
.Iadd.propargyl, .Iaddend.or .[.optionally.]. substituted
propargyl.Iadd.; .Iaddend.and R.sub.3 and R.sub.4 are each
independently .[.hydrogen,.]. C.sub.1-8 alkyl, C.sub.6-12 aryl,
C.sub.6-12 .[.aryl.]. .Iadd.aralkyl .Iaddend.or C.sub.6-12
cycloalkyl.Iadd., .Iaddend.each .Iadd.of which may be
.Iaddend.optionally substituted, .Iadd.or is hydrogen, such
compound being a racemic mixture, an enantiomer, or a salt
thereof;.Iaddend. .[.a racemic mixture, an enantiomer, or salts
thereof,.]. .Iadd.so as .Iaddend.to thereby selectively inhibit
MAO-B activity in the brain of the subject.
11. The method of claim 10 wherein the enantiomer is the R
enantiomer.
12. The method of claim 10 wherein the enantiomer is the S
enantiomer.
13. The method of claim 10 wherein the compound has the structure:
##STR00023##
14. The method of claim 13 wherein the enantiomer is the R
enantiomer.
15. The method of claim 13 wherein the enantiomer is the S
enantiomer.
16. The method of claim 10 wherein the compound has the structure:
##STR00024##
17. The method of claim 16 wherein the enantiomer is the R
enantiomer.
18. The method of claim 16 wherein the enantiomer is the S
enantiomer.
19. A method of treating a subject suffering from depression
comprising administering to the subject a pharmaceutical
composition comprising a therapeutically effective amount of a
compound having the structure: ##STR00025## wherein when a is 0, b
is 1 or 2.Iadd., and when a is 1, b is 1;.Iaddend. .[.wherein when
a is 1, b is 1,.]. m is 0-3.[.,.]. .Iadd.; .Iaddend.X is O or
S.[.,.]. .Iadd.; .Iaddend.Y is halogeno.[.,.]. .Iadd.;
.Iaddend.R.sub.1 is hydrogen or C.sub.1-4 alkyl.[.,.]. .Iadd.;
.Iaddend.R.sub.2 is hydrogen, C.sub.1-4 alkyl, .Iadd.propargyl,
.Iaddend.or .[.optionally.]. substituted propargyl.Iadd.;
.Iaddend.and R.sub.3 and R.sub.4 are each independently
.[.hydrogen,.]. C.sub.1-8 alkyl, C.sub.6-12 aryl, C.sub.6-12
.[.aryl.]. .Iadd.aralkyl .Iaddend.or C.sub.6-12 cycloalkyl, each
.Iadd.of which may be .Iaddend.optionally substituted, .Iadd.or is
hydrogen, such compound being a racemic mixture, an enantiomer, or
a salt thereof .Iaddend..[.a racemic mixture, an enantiomer, or
salt thereof.]. , and a pharmaceutically acceptable carrier.[.,.].
.Iadd.; so as .Iaddend.to thereby treat the subject's
depression.
20. The method of claim 19 wherein the pharmaceutically acceptable
carrier is a solid and the therapeutically effective amount is an
amount from about 0.5 mg to about 2000 mg.
21. The method of claim 19 wherein the pharmaceutically acceptable
carrier is a liquid and the therapeutically effective amount is an
amount from about 0.5 mg to about 2000 mg.
22. The method of claim 19 wherein the pharmaceutically acceptable
carrier is a gel and the therapeutically effective amount is an
amount from about 0.5 mg to about 2000 mg.
23. The method of claim 19 wherein the therapeutically effective
amount is an amount from about 1 mg to about 1000 mg.
24. A method of selectively inhibiting monoamine oxidase-B (MAO-B)
activity in the brain of a subject in need of such inhibition
comprising administering to the subject a pharmaceutical
composition comprising a therapeutically effective amount of a
compound having the structure: ##STR00026## wherein when a is 0, b
is 1 or 2.Iadd., and when a is 1, b is 1.Iaddend.; .[.wherein when
a is 1, b is 1,.]. m is 0-3.[.,.]. .Iadd.; .Iaddend.X is O or
S.[.,.]. .Iadd.; .Iaddend.Y is halogeno.[.,.]. .Iadd.;
.Iaddend.R.sub.1 is hydrogen or C.sub.1-4 alkyl.[.,.]. .Iadd.;
.Iaddend.R.sub.2 is hydrogen, C.sub.1-4 alkyl, .Iadd.propargyl
.Iaddend.or .[.optionally.]. substituted propargyl.Iadd.;
.Iaddend.and R.sub.3 and R.sub.4 are each independently
.[.hydrogen,.]. C.sub.1-8 alkyl, C.sub.6-12 aryl, C.sub.6-12
.[.aryl.]. .Iadd.aralkyl .Iaddend.or C.sub.6-12 cycloalkyl.Iadd.,
.Iaddend.each .Iadd.of which may be .Iaddend.optionally
substituted, .Iadd.or is hydrogen, such compound being a racemic
mixture, an enantiomer, or a salt thereof.Iaddend. .[.a racemic
mixture, an enantiomer, or salt thereof.]. , and a pharmaceutically
acceptable carrier.[.,.]. .Iadd.; so as .Iaddend.to thereby
selectively inhibit MAO-B activity in the brain of the subject.
25. The method of claim 24 wherein the pharmaceutically acceptable
carrier is a solid and the therapeutically effective amount is an
amount from about 0.5 mg to about 2000 mg.
26. The method of claim 24 wherein the pharmaceutically acceptable
carrier is a liquid and the therapeutically effective amount is an
amount from about 0.5 mg to about 2000 mg.
27. The method of claim 24 wherein the pharmaceutically acceptable
carrier is a gel and the therapeutically effective amount is an
amount from about 0.5 mg to about 2000 mg.
28. The method of claim 24 wherein the therapeutically effective
amount is an amount from about 1 mg to about 1000 mg.
.Iadd.29. A compound having the structure: ##STR00027## wherein
when a is 0, b is 1 or 2, and when a is 1, b is 1; m is 0-3; X is O
or S; Y is halogeno; R.sub.1 is hydrogen or C.sub.1-4 alkyl;
R.sub.2 is hydrogen, C.sub.1-4 alkyl, propargyl or substituted
propargyl; and R.sub.3 and R.sub.4 are each independently C.sub.1-8
alkyl, C.sub.6-12 aryl, C.sub.6-12 aralkyl or C.sub.6-12
cycloalkyl, each of which may be optionally substituted, or
hydrogen, such compound being a racemic mixture, an enantiomer, or
a salt thereof..Iaddend.
.Iadd.30. The compound of claim 29, having the structure:
##STR00028## wherein the group OC(O)NR.sub.3R.sub.4 is on the 6
position of the indan ring counting from the amino substituted
carbon atom; m is 0; R.sub.1 is H; R.sub.2 is H; R.sub.3 is methyl;
and R.sub.4 is ethyl..Iaddend.
.Iadd.31. The compound of claim 30, wherein the compound is the R
enantiomer..Iaddend.
.Iadd.32. The compound of claim 29, having the structure:
##STR00029## wherein the group OC(O)NR.sub.3R.sub.4 is on the 6
position of the indan ring counting from the amino substituted
carbon atom; m is 0; R.sub.1 is H; R.sub.2 is H; R.sub.3 is H; and
R.sub.4 is ethyl..Iaddend.
.Iadd.33. The compound of claim 29, having the structure:
##STR00030## wherein the group OC(O)NR.sub.3R.sub.4 is on the 6
position of the indan ring counting from the amino substituted
carbon atom; m is 0; R.sub.1 is H; R.sub.3 is H; and R.sub.4 is
ethyl..Iaddend.
.Iadd.34. The compound of claim 29, wherein the compound is
R-6-(N-methyl, N-ethyl-carbamyloxy)-N'-propargyl-1-aminoindan
hemi-(L)-tartrate..Iaddend.
.Iadd.35. A pharmaceutical composition comprising the compound of
claim 29 and a pharmaceutically acceptable carrier..Iaddend.
.Iadd.36. A pharmaceutical composition comprising the compound of
claim 30 and a pharmaceutically acceptable carrier..Iaddend.
.Iadd.37. A pharmaceutical composition comprising the compound of
claim 31 and a pharmaceutically acceptable carrier..Iaddend.
.Iadd.38. A pharmaceutical composition comprising the compound of
claim 32 and a pharmaceutically acceptable carrier..Iaddend.
.Iadd.39. A pharmaceutical composition comprising the compound of
claim 33 and a pharmaceutically acceptable carrier..Iaddend.
.Iadd.40. A pharmaceutical composition comprising the compound of
claim 34 and a pharmaceutically acceptable carrier..Iaddend.
.Iadd.41. A method of treating a subject suffering from Alzheimer's
disease comprising administering to the subject a therapeutically
effective amount of the pharmaceutical composition of claim 35 so
as to treat the subject..Iaddend.
.Iadd.42. A method of treating a subject suffering from Alzheimer's
disease comprising administering to the subject a therapeutically
effective amount of the pharmaceutical composition of claim 36 so
as to treat the subject..Iaddend.
.Iadd.43. A method of treating a subject suffering from Alzheimer's
disease comprising administering to the subject a therapeutically
effective amount of the pharmaceutical composition of claim 37 so
as to treat the subject..Iaddend.
.Iadd.44. A method of treating a subject suffering from Alzheimer's
disease comprising administering to the subject a therapeutically
effective amount of the pharmaceutical composition of claim 38 so
as to treat the subject..Iaddend.
.Iadd.45. A method of treating a subject suffering from Alzheimer's
disease comprising administering to the subject a therapeutically
effective amount of the pharmaceutical composition of claim 39 so
as to treat the subject..Iaddend.
.Iadd.46. A method of treating a subject suffering from Alzheimer's
disease comprising administering to the subject a therapeutically
effective amount of the pharmaceutical composition of claim 40 so
as to treat the subject..Iaddend.
.Iadd.47. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of the pharmaceutical composition of claim 35 so as to treat
the subject..Iaddend.
.Iadd.48. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of the pharmaceutical composition of claim 36 so as to treat
the subject..Iaddend.
.Iadd.49. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of the pharmaceutical composition of claim 37 so as to treat
the subject..Iaddend.
.Iadd.50. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of the pharmaceutical composition of claim 38 so as to treat
the subject..Iaddend.
.Iadd.51. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of the pharmaceutical composition of claim 39 so as to treat
the subject..Iaddend.
.Iadd.52. A method of treating a subject suffering from depression
comprising administering to the subject a therapeutically effective
amount of the pharmaceutical composition of claim 40 so as to treat
the subject..Iaddend.
.Iadd.53. A method of treating a subject suffering from Attention
Deficit Disorder, Attention Deficit and Hyperactivity Disorder,
Tourette's syndrome, dementia, neurotraumatic disorder or memory
disorder which comprises administering to the subject a
therapeutically effective amount of the pharmaceutical composition
of claim 35 so as to treat the subject..Iaddend.
.Iadd.54. A method of treating a subject suffering from Attention
Deficit Disorder, Attention Deficit and Hyperactivity Disorder,
Tourette's syndrome, dementia, neurotraumatic disorder or memory
disorder which comprises administering to the subject a
therapeutically effective amount of the pharmaceutical composition
of claim 36 so as to treat the subject..Iaddend.
.Iadd.55. A method of treating a subject suffering from Attention
Deficit Disorder, Attention Deficit and Hyperactivity Disorder,
Tourette's syndrome, dementia, neurotraumatic disorder or memory
disorder which comprises administering to the subject a
therapeutically effective amount of the pharmaceutical composition
of claim 37 so as to treat the subject..Iaddend.
.Iadd.56. A method of treating a subject suffering from Attention
Deficit Disorder, Attention Deficit and Hyperactivity Disorder,
Tourette's syndrome, dementia, neurotraumatic disorder or memory
disorder which comprises administering to the subject a
therapeutically effective amount of the pharmaceutical composition
of claim 38 so as to treat the subject..Iaddend.
.Iadd.57. A method of treating a subject suffering from Attention
Deficit Disorder, Attention Deficit and Hyperactivity Disorder,
Tourette's syndrome, dementia, neurotraumatic disorder or memory
disorder which comprises administering to the subject a
therapeutically effective amount of the pharmaceutical composition
of claim 39 so as to treat the subject..Iaddend.
.Iadd.58. A method of treating a subject suffering from Attention
Deficit Disorder, Attention Deficit and Hyperactivity Disorder,
Tourette's syndrome, dementia, neurotraumatic disorder or memory
disorder which comprises administering to the subject a
therapeutically effective amount of the pharmaceutical composition
of claim 40 so as to treat the subject..Iaddend.
Description
FIELD OF INVENTION
The present invention relates to novel compounds, pharmaceutical
compositions containing said compounds and their use in the
treatment of various CNS disorders.
BACKGROUND OF THE INVENTION
Dementia exists in several forms including static dementia,
Alzheimer's-type dementia, senile dementia, pre-senile dementia and
progressive dementia. One of the common pathological features of
several types of dementia is the lack of the neurotransmitter
acetylcholine. This has led to the development of acetylcholine
esterase inhibitors for use in the treatment of dementias such as
the compound tacrine. A summary of the different approaches to and
progress made in the treatment of Alzheimer's Disease may be found
in Drugs of the Future (1995) 20(11): 1145-1162.
Recently, compounds that in addition to inhibiting acetylcholine
esterase, possess inhibitory activity against monoamine oxidase
type A (MAO-A) have been developed. The perceived benefit of having
the anti-MAO-A activity is stated to be an anti-depressant effect
(European Patent Publication Nos. 614,888 and 664,291).
U.S. Pat. Nos. 5,387,133, 5,453,446, 5,457,133 and 5,519,061 all
disclose that the compound (R)-N-propargyl-1-aminoindan, a highly
selective monoamine oxidase type B (MAO-B) inhibitor is effective
in the treatment of dementias of the Alzheimer type and memory
disorders. There is no indication given therein that the compound
might have acetylcholine esterase inhibitory activity. Furthermore,
the compound is only very weakly active as a MAO-A inhibitor.
PCT International Publication No. WO95/18617 discloses various
aminoindan derivatives that are active in a variety of CNS
disorders including dementias of the Alzheimer type. There is no
indication given therein that any of the compounds disclosed might
have acetylcholine esterase inhibitory activity.
SUMMARY OF THE INVENTION
The present invention relates to compounds of formula I
##STR00002## wherein when a is 0; b is 1 or 2; when a is 1, b is 1;
m is from 0 to 3; X is C or S; Y is halogeno; R.sub.1 is hydrogen
or C.sub.1-4 alkyl; R.sub.2 is hydrogen, C.sub.1-4 alkyl or
optionally substituted propargyl; and R.sub.3 and R.sub.4 are each
independently hydrogen, C.sub.1-8 alkyl, C.sub.6-12 aryl,
C.sub.6-12 aralkyl or C.sub.6-12 cycloalkyl optionally
substituted.
The invention relates to the compounds themselves, pharmaceutical
compositions containing said compounds and their use in the
treatment of depression, Attention Deficit Disorder (ADD),
Attention Deficit and Hyperactivity Disorder (ADHD), Tourette's
Syndrome, Alzheimer's Disease and other dementias such as senile
dementia, presenile dementia, progressive dementia, dementia of the
Parkinson's type, vascular dementia and Lewy body dementia.
A further aspect of the present invention relates to the use of the
compounds of formula I in the treatment of neurotraumatic disorder.
As used herein the term "neurotraumatic disorder" is meant to
include damage caused to the nervous system (both central and
peripheral) by virtue of ischemic damage such as that which occurs
in stroke, hypoxia or anoxia, neurodegenerative diseases,
Parkinson's Disease, Alzheimer's Disease, Huntington's Disease,
neurotoxic injury, head trauma injury, spinal trauma injury,
peripheral neuropathy or any form of nerve damage.
An additional aspect of the present invention relates to the use of
the compounds of formula I in the treatment of memory disorder or
depression.
The present invention relates to the racemic compounds themselves
and optically active enantiomers thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the reduction in latency for mice after closed head
injury in the Morris Water Maze Test after treatment with compound
1, compound 10 or Saline (Control) The arrow shows the time off
closed head injury.
FIG. 2 shows the reduction in latency for mice after closed head
injury in the Morris Water Maze Test after treatment with compound
24, compound 25 or Saline (Control) The arrow shows the time of
closed head injury.
FIG. 3 shows the reduction in latency for mice after closed head
injury in the Morris Water Maze Test after treatment with compound
37, compound 39 or Saline (Control). The arrow shows the time off
closed head injury.
DETAILED DESCRIPTION OF THE INVENTION
The present invention is directed to compound of Formula I:
##STR00003## wherein when a is 0, b is 1 or 2; when a is 1, b is :,
m is from 0-3, X is O or S; Y is halogen; R.sub.1 is hydrogen or
C.sub.1-4 alkyl; R.sub.2 is hydrogen, C.sub.1-4 alkyl, or
optionally substituted propargyl and R.sub.3 and R.sub.4 are each
independently hydrogen, C.sub.1-6 alkyl, C.sub.6-11 aryl,
C.sub.6-11; aralkyl or C.sub.6-11 cycloalkyl each optionally
substituted.
In an embodiment of the present invention, a is 0 and b is 1. In
another embodiment of the present invention, a is 0, b is 1, and X
is O.
In an embodiment of the present invention, X is O. In an additional
embodiment of the present invention, X is S.
In an embodiment of the present invention, R.sub.1 is selected from
the group consisting of hydrogen, methyl, ethyl or optionally
substituted propargyl.
In another embodiment of the present invention, R.sub.1 is
propargyl.
In a further embodiment of the present invention, the compound is
selected from the group consisting of: (rac) 6-(N-methyl,
N-ethyl-carbanyloxy)-N'-propargyl-1-aminoindan HCl; (rac)
6-(N,N-dimethyl, carbanyloxy)-N'-methyl-N'-propargyl-1-aminoindan
HCl; (rac) 6-(N-methyl,
N-ethyl-carbamyloxy)-N'-propargyl-1-aminotetralin HCl;
(rac)6-(N,N-dimethyl-thiocarbamyloxy)-1-aminoindan HCl;
(rac)6-(N-propyl-carbamyloxy)-N'-propargyl-1-aminoindan HCl;
(rac)5-chloro-6-(N-methyl,
N-propyl-carbamyloxy)-N'-propargyl-1-aminoindan HCl;
(S)-6-(N-methyl, N-propyl-carbamyloxy)-N'-propargyl-1-aminoindan
HCl; and (R)-6-(N-methyl,
N-ethyl-carbamyloxy)-N'-propargyl-1-aminoindan
hemi-(L)-tartrate.
In a further embodiment of the present invention, R.sub.1 is
hydrogen, methyl or ethyl and R.sub.2 is hydrogen, methyl, ethyl or
optionally substituted propargyl. In a further embodiment of the
present invention, the propargyl group is substituted with a
C.sub.1-.sub.4 alkyl group on the methylene group (R.sub.6 in
Scheme I)
According to the present invention, the term "halogeno" is used to
refer to fluoro, chloro, bromo, or iodo.
In an embodiment of the present invention, when m is greater than 1
each Y may be the same or different.
In an additional embodiment of the present invention, the group
OC(X)NR.sub.3R.sub.4 is on the 4, 6 or 7 position of the indan ring
counting from the amino substituted carbon.
In another embodiment of the present invention, at least one of
R.sub.3 and R.sub.4 is methyl and the other is hydrogen, methyl,
ethyl, propyl, butyl, hexyl, phenyl, benzyl or cyclohexyl.
In the practice of this invention, pharmaceutically acceptable
salts include, but are not limited to, the esylate, mesylate,
maleate, fumarate, tartrate, hemi-tartarate, hydrochloride,
hydrobromide, p-toluenesulfonate, benzoate, acetate, phosphate and
sulfate salts.
The subject invention further provides a pharmaceutical composition
which comprises a therapeutically effective amount of a compound of
formula I or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier. The "therapeutically effective
amount" of a compound of formula I or a pharmaceutically acceptable
salt thereof may be determined according to methods well known to
those skilled in the art, indications of such amounts are given
below.
These compositions may be prepared as medicaments go be
administered orally, parenterally, rectally, or transdermally.
Suitable forms for oral administration include tablets, compressed
or coated pills, dragees, sachets, hard or soft gelatin capsules,
sublingual tablets, syrups and suspensions. In one embodiment, the
pharmaceutically acceptable carrier is a solid and the
pharmaceutical composition is a tablet. The therapeutically
effective amount may be an amount from about 0.5 mg to about 2000
mg, preferably from about 1 mg to about 1000 mg.
In an alternative embodiment, the pharmaceutically acceptable
carrier is a liquid and the pharmaceutical composition is an
injectable solution. The therapeutically effective amount may be an
amount from about 0.5 mg to about 2000 mg, preferably from about 1
mg to about 1000 mg. The volume administered may be an amount
between 0.5 and 10 ml.
In a further alternative embodiment, the carrier is a gel and the
pharmaceutical composition is a suppository. For parenteral
administration the invention provides ampoules or vials that
include an aqueous or non-aqueous solution or emulsion. For rectal
administration there are provided suppositories with hydrophilic or
hydrophobic vehicles. For topical application as ointments and
transdermal delivery there are provided suitable delivery systems
as known in the art. For oral or suppository formulations, 0.5-2000
mg per dosage unit and preferably 1-1000 mg per dosage unit.
These compositions may be used alone to treat the above-listed
disorders, or alternatively, for example, in the case of
Alzheimer's Disease, they may be used as an adjunct to the
conventional treatments such as haloperidol, tacrine or
deprenyl.
The invention will be better understood from the Experimental
Details which follow. However, one skilled in the art will readily
appreciate that the specific methods and results discussed are
merely illustrative of the invention as described more fully in the
claims which follow thereafter.
EXAMPLES
Compounds of general formula I may be prepared, as shown in Scheme
I, from the corresponding carbamoyl derivatives of aminoindan III
by reacting the latter with propargyl compounds bearing an
appropriate leaving group at the 3-position, e.g. a halide group,
mesylate, tosylate, etc., under basic cenditions provided by an
inorganic base, e.g. K.sub.2CO.sub.3, NaOH, or an organic base e.g.
a tertiary amine, in a polar organic solvent, e.g. CH.sub.3CN, DMF,
etc., at 15-40.degree. C., preferably at 20-25.degree. C., for a
period of time in the range of 5-48 hours, preferably 20-30 hours.
The products, obtained after a suitable work-up and purification,
are in the form of free bases. Preferably these are converted into
their pharmaceutically acceptable salts, e.g. HCl, mesylate,
hemi-tartarate, etc.
As shown in Scheme I, compounds of general formula III may be
prepared by Boc deprotection of compounds of general formula IV. In
turn, compounds of general formula IV may be prepared by
carbamylating a compound of general formula V in a conventional
manner, e.g. by reacting the compound of formula V with an
appropriate carbamoyl halogenide or by an alkylisocyanate. Finally,
compounds of general formula V may be prepared by Boc protection of
the appropriate hydroxy amines, by methods known to those skilled
in the art. N,N-dialkyl aminoindan derivatives may be prepared as
shown on in Scheme I by the direct carbamylation of the
corresponding N,N-dialkyl-hydroxy-aminoindan or by alkylation of a
compound of formula III.
Although Scheme I shows the preparation of carbamoyl derivatives
the same process and description above is relevant to the
preparation of the thiocarbamates of the present invention.
Starting Materials
6- and 7-Hydroxy-1-aminoindans may be prepared by demethylation of
the respective 6- and 7-methoxy-1-aminoindans. The latter may be
obtained from the corresponding 1-indanones, either by their
conversion to the oximes, followed by reduction, or by their
reductive amination (NaCNBH.sub.3 and NH.sub.4OAc).sup.2.
6-Hydroxy aminoindan may also be prepared from aminoindan via a
regioselective Friedel--Crafts acylation of a suitably N-protected
aminoindan, followed by a Baeyer--Williger oxidation and finally
hydrolysis.sup.5. 6-hydroxy-(R)-1-aminoindan may thus be prepared
by the method described in the Example below and Scheme II, wherein
"R" is optionally substituted alkyl.
N-Methyl-6-hydroxy-1-aminoindan was prepared by demethylation of
6-methoxy-N-methyl-1-aminoindan, which was prepared from
6-methoxy-1-aminoindan by reductive alkylation (e.g. ethyl formate,
followed by LiAlH.sub.4 reduction), or alternatively, by reductive
amination (MeNH.sub.2, HCl, NaCNBH.sub.3) of
6-methoxy-1-indanone.sup.2. N-ethyl-6-hydroxy-1-aminoindan was
obtained by acetylation of 6-hydroxy-1-aminoindan (Ac.sub.2O, KOH),
followed by reduction (LiAH.sub.4).
N,N-Dimethyl-6-hydroxy-1-aminoindan was prepared by demethylation
of the corresponding 6-methoxy analogue, which was prepared by
reductive alkylation (formaldehyde, formic acid) of
6-methoxy-1-aminoindan. 4-Hydroxy-1-aminoindan may be prepared from
4-hydroxy indanone by converting the latter to the oxime, followed
by reduction.sup.1. 4-Hydroxy indanone may be prepared from
dihydrocoumarin..sup.3
7-Hydroxy-1-aminotetralin and 7-hydroxy-2-aminotetralin were
prepared by demethylation of the corresponding 7-methoxy analogues.
The latter were prepared by reductive amination (as above) of the
corresponding 7-methoxy-1- and 2-tetralones.
7-Methoxy-2-tetralone was prepared from 2,7-dimethoxytetralin
according to Copinga,et al.sup.4.
Preparation of 6-Hydroxy-(R)-1-aminoindan (As Shown in Scheme
II)
N-Trifluoroacetyl-(R)-1-aminoindan
To a cooled (0-5.degree. C.) solution of trifluoroacetic anhydride
(194.6 g, 0.926 mol) in toluene (680 ml) was added dropwise a
solution of (R)-1-aminoindan (base) (113.32 g 0.85 mol) in toluene
(50 ml) and stirred under ice-cooling for 31/2 hours. A solution of
KOH (67.25 g, 1.2 mol) in water (1000 ml) was then added, under
cooling. The reaction mixture was stirred for further 2 hours at
room temperature and filtered. The solid was collected by
filtration, washed with water (680 ml) and dried in vacuo at
60.degree. C. to give 152 g (78%) of a white solid,
mp:153-154.degree. C. The solution was evaporated in vacuum and the
crystals were filtered and washed with water. The solid was dried
in vacuo at 60.degree. C. The second crop (25 g) was crystallized
from a mixture of hexane and ethyl acetate to give 189 (9%) of a
white solid, mp:153-154.degree. C. The total yield was 170 g
(87%).
6-Chloroacetyl-N-trifluoroacetyl-(R)-1-aminoindan
To a suspension of AlCl.sub.3 (89.2 g, 0.67 mol) in
1,2-dichloroethane (600 ml) was added chloroacetyl chloride (55.7
ml, 78.9 g, 0.7 mol) dropwise at 0-5.degree. C. under nitrogen for
20 minutes and it was then left to warm up to 20-25.degree. C. To
this mixture was added N-trifluoroacetyl-(R)-1-aminoindan (34.4 g,
0.15 mol) for 3 hours at 20-25.degree. C. The resulting mixture was
then stirred for an additional 30 minutes and poured into a mixture
of ice-cold water (1.5 l) and 1,2-dichloroethane (1l). The mixture
was stirred for 5 minutes and the layers were separated. The
aqueous layer was extracted with 1,2-dichloroethane (2.times.750
ml). The combined organic layers were washed with water
(2.times.900 ml) and 5% aqueous NaHCO.sub.3 solution (3.times.900
ml). The organic layer was dried (Na.sub.2SO.sub.4) and the solvent
was removed under reduced pressure to give a solid, which was
recrystallized from ethanol to give 15 g (48%) of a white solid mp:
166-167.degree. C.
6-Chloroacetoxyl-N-trifluoroacetyl-(R)-1-aminoindan
6-Choroacetyl-N-trifluoroacetyl-(R)-1-aminoindan (30.57 g, 0.1 mol)
was dissolved in anhydrous dichloromethane (210 ml) and
3-chloroperoxybenzoic acid (70%, 44.87 g, 0.26 mol) was added all
at once. The suspension was cooled to 0.degree. C. and
trifluoroacetic acid (11.4 g, 0.1 mol) was added dropwise for 5-10
minutes. The reaction flask was protected from light and the
mixture was stirred for 3-5 days at room temperature. The reaction
mixture was poured into water (300 ml.). The mixture was
neutralized with ammonium hydroxide solution. The layers were
separated. The aqueous layer was extracted with dichloromethane
(200 ml). The combined organic layers were dried (Na.sub.2SO.sub.4)
and the solvent was removed under reduced pressure to give a solid,
which was recrystallized from ethanol to give 15 g (48%) of a white
solid mp: 169-170.degree. C.
6-Hydroxy-(R)-1-aminoindan
A suspension of 6-chloroacetoxy-N-trifluoroacetyl-(R)-1-aminoindan
(25.4, 0.11 mol) and K.sub.2CO.sub.3 (38.0 g, 0.275 mol) in a
mixture of methanol (275 ml) and water (175 ml) was stirred at
70.degree. C. for 1.5 hours. Methanol was removed in vacuo, and the
aqueous phase was neutralized with 10% hydrochloric acid. The
mixture was filtered and the solid was washed with water. The
mother liquor was evaporated under reduced pressure to a small
volume. The suspension was neutralized, filtered and the brown
solids were crystallized from methanol (twice) to give 7.0 g (43%)
of a white solid mp:200-203.degree. C.
Preparation of the corresponding S-enantiomer may be carried out in
the same manner using (S)-1-aminoindan as the starting
material.
Resolution of Enantiomers
The R- and S-enantiomers of each compound may be obtained by
optical resolution of the corresponding racemic mixtures. Such a
resolution can be accomplished by any conventional resolution
method well known to a person skilled in the art, such as those
described in U.S. Pat. No. 4,833,273, issued May 23, 1989 (Goel)
and in J. Jacques, A. Collet and S. Wilen, "Enantiomers, Racemates
and Resolutions," Wiley, N.Y. (1981). For example, the resolution
may be carried out by preparative chromatography on a chiral
column. Another example of a suitable resolution method is the
formation of diastereomeric salts with a chiral acid such as
tartaric, malic, mandelic acid or N-acetyl derivatives of amino
acids, such as N-acetyl leucine, followed by recrystallization to
isolate the diastereomeric salt of the desired enantiomer.
Alternatively, selected starting materials, intermediates or end
products may be resolved into their respective enantiomers by the
method described in PCT International Application Publication No.
WO/96US/21640, wherein the compound to be resolved is first
converted into its N-benzyl derivative. The N-benzyl derivative is
then resolved using either R or S-mandelic acid. The resolved
product is converted to its base and reduced under acidic
conditions to provide the desired enantiomer. Preferably, the
starting material is resolved prior to Boc protection and
carbamylation.
The R and S enantiomers of the starting materials may also be
prepared from R and S enantiomers c: aminoindan via a
regioselective Friedel--Crafts acylation so a suitably N-protected
optical isomer of aminoindan, followed by a Baeyer-Williger
oxidation and finally hydrolysis.sup.5, thus obviating the need for
optical resolution.
REFERENCES
1. Y. Oshiro, et al, J. Med. Chem. 34: 2004 (1991); 2. R. F. Borch,
et al, J. Am. Chem. Soc. 93:, 2897 (1971); 3. J. G. Cannon, et al,
J. Med. Chem. 28: 515 (1985); 4. S. C. Copinga, et al, J. Med.
Chem. 36: 2891 (1993); and 5. K. Teranishi et al, Synthesis 1018
(1994). Preparation of Compounds of the Invention as Shown in
Scheme I
A: Boc--protection and carbamylation
1. Boc Protection
6-hydroxy N-Boc aminoindan
A solution of 6-hydroxy aminoindan (16 g, 107 mmol), di-t-butyl
dicarbonate (23.8 g, 109.2 mmol) and Et.sub.3N (16.74 ml, 120 mmol)
in THF (375 ml) was stirred at room temperature (RT) for 20 hrs.
The reaction mixture was evaporated to dryness under reduced
pressure, and the residue was dissolved in CH.sub.2Cl.sub.2 (200
ml), washed with water (200 ml), dried over Na.sub.2SO.sub.4 and
evaporated to dryness under reduced pressure. The crude product was
purified by column chromatography (hexane/EtOAc 2:1) to give 23 g
of a solid (86%).
2. Carbamylation
6-(N-Me, N-Et carbamyloxy) N-Boc aminoindan
To a stirred and ice-cooled solution of N-Boc 6-hydroxy aminoindan
(7.5 g, 30 mmol) in acetonitrile (75 ml) was added N-Me,N-Et
carbamoyl chloride (6.3 g, 51.8 mmol), followed by a dropwise
addition of NaH (60% in oil, 1.56 g, 39 mmol). The reaction mixture
was stirred for 2 hrs at RT under argon. After evaporation of the
solvent in-vacuo, water (100 ml) was added, and extracted with
ether (3.times.100 ml). The organic phase was washed with dilute
NaOH (pH 10-11), dried and evaporated to dryness in-vacuo.
Purification by column chromatography (hexane:EtOAc 2:1) afforded
7.8 g (77%) of an oil.
In this manner the intermediates in Tables 1 and 2 were prepared.
In Table 1 and all further Tables the heading "position" refers to
the ring position of the carbamyl group unless otherwise
indicate
TABLE-US-00001 TABLE 1 N-Boc protected carbomyloxy aminoindans
##STR00004## position Y R1 R3 R4 yield (%) 6- H H Me Me 92 6- H H
Me Pr 95 6- H H Me Et 77 7- H H Me Me 92 7- H H Me Et 83 7- H H Me
Pr 95 6- H Et Me Me 76 6- H Me Me Me 92 7- H Me Me Me 78 6- H Me Me
Pr 80 6- H H Me n-hexyl 98 4- H H Me Me 85 4- H H Me Et 87 6- H H
Me Et 89 6- H H Me cyclohexyl 98 6- H H Me p-OMe-phenyl 97 6- H H
Me phenyl 93 6- H H Me CH.sub.2-phenyl 83 6- 5-Cl H Me Et 88 6-
5-Cl H Me Pr 97 6- H H Me Bu 99 6- H H Et Bu 93 6- H H Et
cyclohexyl 94
TABLE-US-00002 TABLE 2 N-Boc protected carbomyloxy tetralins
##STR00005## position of amine R1 R3 R4 yield (%) 2- H Me Me 85 2-
H Me Et 79 1- H Me Me 85 1- H Me Et 98
B: Boc--Deprotection
6-(N-Me,N-Et Carbamyloxy) aminoindan HCl (Compound 3)
6-(N-Me,N-Et Carbamyloxy) N-Boc aminoindan (7.8 g, 23.3 mmol) was
dissolved in dioxane (80 ml), and a 20% solution of gas. HCl in
dioxane (80 ml) was added. After 2 hr stirring at RT the solvent
was evaporated in-vacuo and the residue was treated with dry ether
(200 ml) and the mixture stirred at RT for 4 hrs and filtered, to
give 6.15 g (0.7 mmol, 97%) of 6-(N-Me, N-Et carbamyloxy)
aminoindan hydrochloride.
In this manner the following compounds of general formula I as
shown in Tables 3, 3a and 4 were prepared. Spectral data relating
to these compounds is given in Tables 7, 7a and 8.
TABLE-US-00003 TABLE 3 Carbamyloxy aminoindan HCl salts
##STR00006## cryst/ slurry yield # position R1, R2 R3 R4 solvent
mp(.degree. C.) (%) 1 6- H, H Me Me Et.sub.2O 156-8 93 2 6- H, H Me
Pr Et.sub.2O 165-7 27 3 6- H, H Me Et Et.sub.2O 150-2 50 4 7- H, H
Me Me Et.sub.2O 156-60 93 5 7- H, H Me Et Et.sub.2O 185-7 55 6 7-
H, H Me Pr Et.sub.2O 153-5 33 7 6- H, Et Me Me Et.sub.2O 172-4 91 8
6- H, Me Me Me Et.sub.2O 178-80 88 9 7- H, Me Me Me dioxane 169-71
98 10 6- H, Me Me Et Et.sub.2O 172-4 87 11 6- H, Me Me Pr Et.sub.2O
165-7 98 12 6- Me, Me Me Me Et.sub.2O 164-6 62 13 4- H, H Me Me
Et.sub.2O 198-200 90 14 4- H, H Me Et Et.sub.2O 183-5 92 15 6- H, H
Me n- dioxane 111-12 78 hexyl 16* 6- H, H Me Et Et.sub.2O 197-8 89
17 6- H, H Me cyclo Et.sub.2O 207-8 86 hexyl 18** 6- H, H Me Et
Et.sub.2O 202-4 84 48 6- H, H H Et MeOH/ 191-2 74 EtOAc 49 6- H, H
H Pr MeOH/ 171-3 67 EtOAc 50 6- H, H Me p-OMe- iPrOH 225-7 92
Phenyl 51 6- H, H Me CH.sub.2-- Et.sub.2O 78 Ph 52* 6- H, H Me Me
Et.sub.2O 83 53** 6- H, H Me Me Et.sub.2O 81 88 6- H, H Me Ph
Et.sub.2O 96 66*** 6- H, H Me Et Et.sub.2O 116-9 92 67*** 6- H, H
Me Pr Et.sub.2O 181-3 86 80 6- H, H Me Bu Et.sub.2O 54 84 6- H, H
Et cyclo- Et.sub.2O 196-8 89 hexyl *R < nantiomer **S-cnantiomer
***5-chloro
TABLE-US-00004 TABLE 3a Thiocarbamyloxy aminoindan HCl salts
##STR00007## cryst/slurry yield # position R1, R2 R3 R4 solvent
mp(.degree. C.) (%) 44 6- H, H Me Me MeOH/EtO 244-5 55 45 6- H, H
Me Et MeOH/EtOAc 236-8 58
TABLE-US-00005 TABLE 4 Carbamyloxy aminotetralin HCl salts
##STR00008## position of cryst/slurry yield # amine R1 R3 R4
solvent mp(.degree. C.) (%) 19 2- H Me Me ether a) 96 20 2- H Me Et
ether a) 98 21 1- H Me Me ether 196-8 99 22 1- H Me Et ether 166-8
85 a) wide melting range; compound is a hemi-hydrate
C: Propargylation and salt formation
The compounds prepared in Step B may be optionally propargylated to
provide further compounds of general formula I.
6-(N-Me, N-Et carbamyloxy) N-propargyl aminoindan, HCl (Compound
25)
To a stirred mixture of 6-(N-Me, N-Et carbamyloxy) aminoindan. HCl
(5.2 g, 19.2 mmol), potassium carbonate (5.31 g, 38.4 mmol) in
acetonitrile (250 ml), was added a solution of propargyl bromide
(2.06 g, 17.28 mmol) in acetonitrile (10 ml). The reaction mixture
was stirred at RT under nitrogen for 25 hrs, and filtered. The
filtrate was evaporated to dryness in-vacuo and the residue was
purified by column chromatography (EtOAc) to give 3.6 g (13.2 mmol,
69%) of the free base as a yellow oil.
The free base was dissolved in dry ether (150 ml) and HCl/ether (15
ml) was added. The mixture was stirred at RT for 1 hr, filtered and
the solid was recrystallized from iPrOH/ether to give 3.5 g (11.3
mmol, 59%) of the title compound as a white solid.
6-(N,N-Dimethylcarbamyloxy)-N-propargyl aminoindan mesylate
(Compound 24)
To a stirred mixture of 6-(N,N-dimethylcarbamyloxy) aminoindan HCl
(1.88 g, 7.33 mmol), K.sub.2CO.sub.3 (2.03 g, 14.66 mmol) and
acetonitrile (70 ml) was added a solution of propargyl bromide
(0.79 g, 6.6 mmol) in CH.sub.3CN (5 ml) dropwise over 5 min, under
nitrogen. The mixture was stirred under N.sub.2 for 24 hrs,
filtered and the solvent was removed at reduced pressure. The
residue was taken up into water (150 ml) and toluene (150 ml). This
mixture was stirred while adjusting the pH of the aqueous layer to
3.75 by the addition of 20% aq. HCl. The aqueous layer was
separated and extracted with toluene (2.times.100 ml) and brought
carefully to pH 7.5 by the addition of 10% aq. NaOH solution. It
was then extracted with toluene (100 ml+4.times.70 ml). The
combined toluene layers were dried (Na.sub.2SO.sub.4), filtered and
the solvent was removed under reduced pressure to give 1.06 g (62%)
of a yellow oil.
To a stirred solution of the free base (1.65 g, 6.4 mmol) in anh.
ether (60 ml) was added dropwise a solution of methanesulfonic acid
(0.7 g, 7.29 mmol) in ether (10 ml). The resulting suspension was
stirred at 25.degree. C. for 30 man and then allowed to settle for
an additional 30 min. The ether was then decanted off, and the
residue was dried under vacuum. It was then recrystallized from
iPrOH/ether to give 2.05 g of a white solid (90.3%).
In this manner the following compounds of general formula I as
shown in Tables 5, 5a and 6 were prepared. Analytical data relating
to these compounds is given in Tables 9, 9a and 10.
TABLE-US-00006 TABLE 5 Carbamyloxy-N-propargyl aminoindans
##STR00009## cryst/slurry mp yield # X position R1 R3 R4 solvent
(.degree. C.) (%) 23 Cl 6- H Me Me iPrOH/Et.sub.2O 180-2 52 24
mesylate 6- H Me Me iPrOH/Et.sub.2O 147-9 60 25 Cl 6- H Me Et
iPrOH/Et.sub.2O 194-6 59 26 Cl 6- H Me Pr iPrOH/Et.sub.2O 183-5 46
27 Cl 7- H Me Me iPrOH/Et.sub.2O 219-20 65 28 Cl 7- H Me Pr
iPrOH/Et.sub.2O 185-6 53 29 Cl 6- Me Me Me iPrOH/Et.sub.2O 199-201
55 30 Cl 6- Me Me Et Et.sub.2O 196-8 47 31 Cl 6- Et Me Me
iPrOH/Et.sub.2O 212-3 71 32 Cl 7- Me Me Me iPrOH/Et.sub.2O 169-71
63 33 Cl 7- H Me Et iPrOH/Et.sub.2O 208-9 64 34 Cl 4- H Me Me
Et.sub.2O 196-8 85 35 Cl 4- H Me Et Et.sub.2O 183-5 85 36 Cl 6- H
Me n-hexyl iPrOH/Et.sub.2O 106-8 53 37* Cl 6- H Me Et Et.sub.2O
159-6 88 38 Cl 6- H Me cyclohexyl Et.sub.2O 174-5 55 39** Cl 6- H
Me Et Et.sub.2O 160-2 61 54* mesylate 6- H Me Me Et.sub.2O 139-41
54 55** mesylate 6- H Me Me Et.sub.2O 138-40 52 56 Cl 6- H H Et
iPrOH/Et.sub.2O 175-7 38 57 Cl 6- H H Pr iPrOH/Et.sub.2O 165-7 48
58 mesylate 6- H Me Et Et.sub.2O 92-4 64 59** mesylate 6- H Me Et
iPrOH/Et.sub.2O 72 60 mesylate 6- H Me Et Et.sub.2O 121-3 87 61 Cl
6- H Me p-OMe-Ph Et.sub.2O 172-4 84 62 Cl 6- H Me Ph Et.sub.2O
182-4 61 63 Cl 6- H Me CH.sub.2Ph Et.sub.2O 188-90 58 64*** Cl 6- H
Me Me iPrOH/Et.sub.2O 195-7 55 65*** Cl 6- H Me Et iPrOH/Et.sub.2O
188-90 51 68**** fumarate 6- H Me Et iPrOH 146-8 48 69* fumarate 6-
H Me Et iPrOH 115-7 35 70 crylate 6- H Me Et EtOAc 109-11 60 71****
Cl 6- H Me Et Et.sub.2O 161-3 55 72**** Cl 6- H Me Pr Et.sub.2O
164-6 58 73** fumarate 6- H Me Et iPrOH 114-6 81 74** crylate 6- H
Me Et EtOAc 95-7 82 75** .[.1/2D-tartrale.].
.Iadd.1/2D-tartrate.Iaddend. 6- H Me Et iPrOH 143-5 44 76*
.[.1/2L-tarate.]. .Iadd.1/2L-tartrate.Iaddend. 6- H Me Et iPrOH
143-5 41 77* crylate 6- H Me Et EtOAc 106-8 93 78* Cl 6- H Me Pr
Et.sub.2O 126-8 89 79* Cl 6- H Me Pr Et.sub.2O 135-7 33 81 Cl 6- H
Me Bu Et.sub.2O 168-70 63 83 Cl 6- H Et Bu Et.sub.2O 148-50 42 85
Cl 6- H Et cyclohexyl Et.sub.2O 178-80 56 86* Cl 6- H Me Bu
Et.sub.2O 86-8 51 87** Cl 6- H Me Bu Et.sub.2O 88-9 52
*R-enantiomer **S-enantiomer ***substituted propargyl derivatives,
R.sub.n in Scheme 1 is methyl ****Y: 5-Cl
TABLE-US-00007 TABLE 5a Thiocarbamyloxy-N-propargyl aminoindans
##STR00010## cryst/slurry mp yield # X position R1 R3 R4 solvent
(.degree. C.) (%) 46 Cl 6- H Me Me Et.sub.2O 152-4 53 47 Cl 6- H Me
Et Et.sub.2O 193-5 54
TABLE-US-00008 TABLE 6 N-Propargyl aminotetralins ##STR00011##
position of cryst/slurry mp yield # amine R1 R3 R4 solvent
(.degree. C.) (%) 40 2- H Me Me MeOH/Et.sub.2O 206-8 66 41 2- H Me
Et iPrOH/Et.sub.2O 208-9 65 42 1- H Me Me ether 207-9 57 43 1- H Me
Et ether 201-3 42
TABLE-US-00009 TABLE 7 Analytical Data of Compounds of the
Invention shown in Table 3 ##STR00012## NMR.sub.1 MS elem. anal. #
aryl index R1, R2 R3, R4 IR (MH*) (C, H, N) 1 7.38, 7.20 4.85, 3.10
3.10, 2.96 3446, 2943 221 calc: 56.14, 6.62, 10.90 7.10 2.96, 2.63
1711, 1487 found: 55.90, 6.67, 10.89 2.14 1393, 1240 2 7.40, 7.21
4.80, 3.10 3.43, 3.27 2970, 2863 249 calc: 59.05, 7.38, 9.84 7.10
2.95, 2.65 3.10, 2.95 1735, 1608 found: 58.75, 7.33, 9.86 2.15
1.70, 1.63 1396, 1241 0.94, 0.90 2a 7.40, 7.21 4.80, 3.10 3.43,
3.27 2970, 2863 249 calc: 57.23, 7.55, 9.54 (1/2 7.10 2.95, 2.65
3.10, 2.95 1735, 1608 found: 57.54, 7.29, 9.45 H.sub.2O 2.15 1.70,
1.63 1396, 1241 0.94, 0.90 4 7.47, 7.36 4.91, 3.25 3.18, 3.03 2950,
1701 7.09 3.07, 2.60 1504, 1396 2.25 1234, 1177 5 7.44, 7.29 4.88,
3.20 3.55, 3.39 3446, 2920 235 calc: 57.70, 7.25, 10.35 7.02 3.14,
2.55 3.14, 2.99 1710, 1472 found: 57.38, 6.97, 10.32 2.23 1.26,
1.18 1403, 1235 6 7.45, 7.30 4.86, 3.20 3.50, 3.32 3448, 2923 249
calc: 59.05, 7.43, 9.84 7.02 3.04, 2.55 3.13, 2.98 1710, 1485
found: 58.78, 7.47, 9.91 2.23 1.70, 1.63 1226, 1154 0.94, 0.90 7
7.45, 7.29 4.83, 3.17 3.20, 1.33 3.15, 3.0 2948, 2766 249 calc:
59.05, 7.38, 9.84 7.17 3.02, 2.65 2680, 1725 found: 57.75, 7.40,
9.65 1485, 1386 8 7.43, 7.27 4.75, 3.14 2.73 3.13, 2.97 2950, 2722
235 calc: 57.70, 7.02, 10.35 7.17 3.03, 2.60 1720, 1390 found:
56.83, 7.09, 10.27 2.30 1160 9 7.52, 7.37 4.83, 3.27 2.74 3.19,
3.04 2963, 2710 235 calc: 57.70, 7.02, 10.35 7.10 3.10, 2.55 1715,
1579 found: 57.46, 6.73, 10.36 2.38 1472, 1389 10 7.44, 7.29 4.80,
3.15 2.74 3.55, 3.35 2950, 2705 calc: 59.08, 7.38, 9.84 7.15 3.03,
2.62 3.12, 2.98 1720, 1450 found: 58.74, 7.51, 9.72 2.30 1.25, 1.18
1402 11 7.42, 7.25 4.75, 3.15 2.72 3.45, 3.30 2963, 2723 calc:
60.33, 7.70, 9.38 7.14 3.10, 2.60 3.10, 2.95 1715, 1465 found:
60.32, 7.75, 9.42 2.28 1.65, 0.94 1404, 1234 0.88 12 7.43, 7.27
4.96, 3.12 2.75 3.10, 2.96 3480, 1718 249 calc: 59.05, 7.38, 9.84
7.17 3.05, 2.55 1475, 1390 found: 58.75, 7.41, 9.84 2.42 1237, 1174
13.sup.11 7.53, 7.29 4.71, 2.95, 8.75 3.04, 2.9 221 7.08 2.74,
2.45, 2.0 14.sup.11 7.53, 7.3, 4.71, 2.95, 8.7 3.41, 3.3, 235 7.08
2.73, 2.48, 3/01, 2.89, 2.0 1.18, 1.07 15 7.35, 7.23 4.83, 3.3 3.1,
3.06 2930, 1720 291 calc: 62.47, 8.33, 8.57 7.01 2.6, 2.16 2.95,
2.91 1471, 1405 found: 62.54, 8.30, 8.61 1.6, 1.29 1248 0.85 16
7.42, 7.22 4.87, 3.16 3.53, 3.39 235 7.12 3.01, 2.65 3.92, 2.99
2.17 1.26, 1.17 17 7.42, 7.22 4.87, 3.15 4.10, 3.85 289 calc:
62.85, 7.76, 8.63 7.11 2.95, 2.65 3.00, 2.85 found: 62.55, 7.81,
8.33 2.17 1.90- 1.40 1.34, 1.13 3 7.43, 7.20 4.86, 3.15 3.51, 3.38
235 calc: 55.70, 7.25, 10.35 7.12 3.02, 2.64 3.10, 2.95 found:
57.44, 7.06, 10.38 2.18 1.25, 1.15 18 7.43, 7.20 4.86, 3.15 3.51,
3.38 235 calc: 55.70, 7.25, 10.35 7.12 3.02, 2.64 3.10, 2.95 found:
57.44, 7.06, 10.38 2.18 1.25, 1.35 48 7.41, 7.24 4.87, 3.13 3.23,
1.17 221 calc: 56.13, 6.68, 10.91 7.13 3 0, 2.65 found: 56.00,
6.66, 10.81 2.17 49 7.41, 7.24 4.87, 3.12 3.17, 1.56 235 calc:
57.67, 7.07, 10.35 7.13 2.98, 2.65 0.94 found: 57.32, 7.13, 10.31
2.17 50 7.37, 7.16 4.80, 3.10 7.40- 7.0 calc: 61.98, 6.02, 8.03
7.03 2.96, 2.61 3.82, 3.43 found: 61.16, 6.07, 7.77 2.15 3.29 66
7.57, 7.39 4.91, 3.18 3.61, 3.43 269 calc: 50.41, 6.02, 9.05 3.05,
2.71, 3.20, 3.03 271 found: 50.46, 6.11, 8.77 2.25 1.33, 1.23 67
7.55, 7.36 4.89, 3.14 3.52, 3.36 283 calc: 52.67, 6.32, 8.78 3.02,
2.68 3.18, 3.02 285 found: 52.67, 6.28, 8.48 2.20 1.77, 1.67 0.99,
0.93 .sup.1D.sub.2O, unless otherwise specified
.sup.11DMSO-d.sub.6
TABLE-US-00010 TABLE 7a Analytical Data of Compounds of the
Invention shown in Table 3a ##STR00013## NMR(D.sub.3O) MS elem.
anal. # aryl indan R1, R2 R3, R4 IR (MH.sup.+) (C, H, N, S) 44
7.45, 7.20, 4.87, 3.15, 3.44, 3.36 2933, 1714, calc: 52.83, 628,
10.27, 11.75 7.11 3.05, 2.65, 1599, 1536, found: 51.11, 6.48,
10.23, 12.16 2.20 1488, 1392 45 7.45, 7.20, 4.75, 3.10, 3.88, 3.79
2934, 1719, calc: 51.22, 6.94, 9.19, 10.52 7.11 2.97, 2.65, 3.39,
3.32, 1594, 1522, found: 51.04, 7.30, 9.31, 11.24 2.20 1.28, 1.25
1497, 1402
TABLE-US-00011 TABLE 8 Analytical Data of Compounds of the
Invention shown in Table 4 ##STR00014## NMR.sup.2 MS elem. anal. #
aryl cyclohex. R1, R2 R3, R4 IR (MH.sup.+ (C, H, N) 19 7.22, 6.95
3.69, 3.22 3.12, 2.97 3484, 2930 235 calc: 55.81, 7.20, 10.02
(1/2H.sub.2O) 2.93, 2.87 2362, 1699 found: 55.29, 6.93, 9.71 2.22,
1.92 1612, 1500 1391 20 7.20, 6.94 3.70, 3.19 3.48, 3.35 249 calc:
57.23, 7.55, 9.54 (1/2H.sub.2O) 2.90, 2.23 3.08, 2.94 found: 57.50,
7.53, 9.54 1.90 1.20, 1.12 21 7.28, 7.11, 3.10, 2.96 3.10, 2.96 235
calc: 57.70, 7.02, 10.35 7.06 2.77, 2.16 found: 56.97, 6.93, 10.06
2.05, 1.88 22 7.29, 7.13, 4.57, 2.88 3.52, 3.37 249 calc: 59.05,
7.38, 9.84 7.07 2.79, 2.15 3.10, 2.97 found: 58.91, 7.18, 9.99
2.05, 1.90 1.25, 1.17 .sup.2D.sub.2O, unless otherwise
specified
TABLE-US-00012 TABLE 9 Analytical Data of Compounds of the
Invention shown in Table 5 ##STR00015## NMR.sup.3 MS elem. anal. #
aryl indan R1 proparg R3, R4 IR (MH.sup.+) (C, H, N) 23 7.46, 7.30
5.01, 3.20 4.0, 3.16 3.15, 3.0 259 calc: 61.12, 6.50, 9.51 7.18
3.15, 2.65 found: 60.93, 6.38, 9.47 2.36 24 7.46, 7.30 5.01, 3.20
4.0, 3.16 3.15, 3.02 1711, 1482, 259 calc: 54.22, 6.26, 7.91 7.18
3.15, 2.65 1439, 1394, found: 53.92, 6.28, 7.84 2.36 1192, 1170 25
7.42, 7.27 4.97, 3.16 3.97, 3.02 3.52, 3.36 1728, 1435, 273 calc:
62.23, 6.86, 9.57 7.15 3.0, 2.62 3.10, 2.97, 1403, 1242, found:
62.42, 6.84, 8.94 2.32 1.24, 1.15 1166 25.sup.a 7.50, 7.32 4.78,
3.10 3.91, 3.74 3.43, 3.32 1728, 1435, 273 calc: 62.23, 6.86, 9.57
7.10 2.85, 2.45 3.03, 2.90 1403, 1242, found: 62.42, 6.84, 8.94
2.28 1.20, 1.10 1166 26 7.45, 7.30 5.0, 3.16 4.0, 3.03 3.48, 3.32
1725, 1465, 287 calc: 63.25, 7.18, 8.68 7.17 3.04, 2.65 3.12, 2.98
1429, 1403, found: 63.13, 7.28, 8.93 2.33 1.72, 1.63 1232, 1165
0.96, 0.92 27 7.52, 7.38 5.05, 3.26 3.90, 3.21 3.12, 3.03 3200,
1722, 259 calc: 61.12, 6.50, 9.51 7.10 3.07, 2.56 1567, 1434,
found: 61.01, 6.46, 9.64 2.40 1408, 1238 28 7.52, 7.37 5.02, 3.27
3.98, 3.10 3.65, 3.42 3200, 1727, 287 calc: 63.25, 7.18, 8.68 7.07
3.09, 2.55 3.18, 3.02 1566, 1468, found: 63.06, 7.30, 8.37 2.38
1.75, 0.98 1438, 1406, 0.93 1222 29 7.44, 7.30 5.20, 3.15 2.80
4.01, 3.13 3.12, 2.97 1729, 1388, 273 calc: 62.33, 6.80, 9.07 7.19
3.03, 2.57, 1234, 1165 found: 61.97, 6.80, 8.78 2.44 31 7.48, 7.30
5.34, 3.20 3.36, 4.05, 3.12 3.16, 3.01 3180, 1723, 287 calc: 63.25,
7.18, 8.68 7.23 3.08, 2.65 1.37 1490, 1440, found: 63.42, 7.09,
8.71 2.50 1389, 1230, 1160 32 7.56, 7.39 5.30, 3.28 2.78 4.12, 3.23
3.20, 3.02 1712, 1472, 273 calc: 62.23, 6.86, 9.07 7.15 3.09, 2.55
1392, 1238, found: 62.05, 6.81, 8.87 1171 33 7.46, 7.32 4.96, 2.50
3.92, 3.04 3.13, 2.96 1719, 1426, 273 calc: 62.23, 6.86, 9.07 7.03
2.33 1.24, 1.15 1404, 1233, found: 62.19, 6.77, 9.08 1154 34 7.48,
7.23 5.07, 3.08 4.05, 3.07 3.29, 3.03 3238, 2907 259 calc: 2.95,
2.65 2769, 2635 found: 2.35 1714, 1470 1392, 1240 35 7.48, 7.23
5.07, 3.08 4.05, 3.07 3.56, 3.41 3197, 2934 273 calc: 2.95, 2.65
3.15, 3.01 2565, 2431 found: 2.35 1.29, 1.21 1707, 1445 1403, 1236
36 7.45, 7.28 4.98, 3.16 3.98, 3.04 3.49, 3.35 calc: 65.83, 8.01,
7.68 7.15 3.03, 2.63 3.11, 2.97 found: 65.65, 8.11, 7.82 2.33 1.66,
1.33 0.88 37 7.44, 7.29 4.98, 3.15 3.98, 3.03 3.53, 3.38 3275, 2754
273 calc: 62.23, 6.86, 9.07 7.18 3.01, 2.63 3.12, 2.98 1719, 1445
found: 62.30, 6.94, 9.09 2.31 1.25, 1.16 1395, 1303 38 7.44, 7.27
4.98, 3.14 3.98, 3.04 4.09, 3.85 3227, 2936 327 calc: 66.19, 7.50,
7.72 7.16 3.00, 2.64 3.01, 2.88 2612, 2128 found: 65.90, 7.63, 7.55
2.33 1.90-1.45 1713, 1584 1.35, 1.14 1440, 1401 39 7.46, 7.30 4.97,
3.17 3.97, 3.03 3.54, 3.39 3275, 2933 273 calc: 62.23, 6.86, 9.07
7.19 3.04, 2.64 3.13, 3.0 2758, 1720 found: 62.27, 6.95, 9.03 2.32
1.27, 1.19 1442, 1396 1303 54 7.46, 7.30 5.00, 3.17 3.99, 3.05
3.15, 3.0 1711, 1482 259 calc: 54.17, 6.20, 7.90 7.19 3.05, 2.64
1438, 1395 found: 54.18, 6.27, 7.78 2.33 1192, 1169 55 7.46, 7.30
5.00, 3.17 3.99, 3.05 3.15, 3.0 1711, 1482 259 calc: 54.17, 6.20,
7.90 7.19 3.05, 264 1438, 1395 found: 54.07, 6.25, 7.88 2.33 1192,
1169 56 7.46, 7.32 4.99, 3.17 3.99, 3.05 3.27, 1.20 259 calc:
61.12, 6.50, 9.51 7.20 3.04, 2.65 found: 60.87, 6.47, 9.34 2.33 57
7.47, 7.32 4.99, 3.18 3.99, 3.06 3.20, 1.61 273 calc: 62.23, 6.86,
9.07 7.20 3.05, 2.65 0.98 found: 61.60, 6.93, 9.04 2.34 58 7.47,
7.32 5.01, 3.20 4.01, 3.07 3.56, 3.41 273 calc: 55.43, 6.52, 7.60
7.22 3.08, 267 3.14, 3.01 found: 55.08, 6.52, 7.31 2.36 1.29, 1.21
59 7.47, 7.32 5.01, 3.20 4.01, 3.07 3.56, 3.41 273 calc: 7.22 3.08,
2.67 3.14, 3.01 found: 2.36 1.29, 1.21 60 7.47, 7.32 5.01, 3.20
4.01, 3.07 3.56, 3.41 273 calc: 55.43, 6.52, 7.60 7.22 3.08, 2.67
3.14, 3.01 found: 55.21, 6.64, 7.40 2.36 1.29, 1.21 61 7.40-7.0
4.96, 3.10 3.96, 3.90 7.40-7.0 351 calc: 65.20, 5.95, 7.24 2.97,
2.57 3.03 3.81 found: 64.72, 6.04, 6.81 2.30 62 7.60-7.10 4.96,
3.15 3.98, 3.07 7.60-7.10 321 calc: 67.32, 5.89, 7.85 3.00, 2.61
3.42 found: 67.22, 6.00, 7.54 2.34 63 7.55-7.10 4.97, 3.17, 3.99,
3.07 7.55-7.10 335 calc: 67.47, 6.20, 7.55 3.00, 2.64, 4.73, 4.59
found: 67.75, 6.32, 7.47 2.36, 2.36 3.14, 3.05 64 7.48, 7.35 5.16,
5.12 4.44, 4.27 3.17, 3.03 273 calc: 62.23, 6.86, 9.07 7.21 3.20,
3.05 3.17, 1.68 found: 62.22, 6.86, 8.96 2.70, 2.35 1.63 65 7.44,
7.36, 5.15, 5.09 4.43, 4.25 3.55, 3.39 calc: 63.25, 7.18, 8.68
7.27, 7.19 3.20, 3.02 3.25, 3.17 3.13, 3.00 found: 63.15, 7.15,
8.31 2.65, 2.32 1.67, 1.61 1.27, 1.19 71 7.60, 7.44 5.02, 3.20,
4.02, 3.07 3.60, 3.43, 307 calc: 55.98, 5.87, 8.16 3.06, 2.68,
3.20, 3.02, 309 found: 55.72, 5.88, 8.11 2.36 1.33, 1.23 72 7.59,
7.44 5.01, 3.20, 4.03, 3.07 3.53, 3.36, 321 calc: 57.15, 6.21, 7.84
3.06, 2.68, 3.20, 3.02, 323 found: 57.05, 6.21, 7.81 2.38 1.79,
1.68, 1.01, 0.95 76 7.47, 7.31, 5.00, 3.20, 4.00, 3.07 3.56, 3.40,
3286, 2972, 273 calc: 62.17, 6.62, 8.05 7.20 3.06, 2.66, 3.16,
3.00, 1724, 1637, found: 62.31, 6.66, 7.94 2.35 1.28, 1.20 1400,
1308, 1233 81 7.48, 7.31, 5.00, 3.20, 4.01, 3.07 3.53, 3.38, calc:
64.19, 7.42, 8.32 7.20 3.07, 2.66, 3.14, 3.01, found: 63.99, 7.42,
8.04 2.35 1.65, 1.39, 0.97 83 7.47, 7.31, 5.00, 3.19, 4.01, 3.07
3.52, 3.38, 315 calc: 65.04, 7.70, 7.98 7.19 3.04, 2.66, 1.68,
1.40, found: 64.75, 7.72, 7.94 2.34 1.29, 1.22, 0.98 85 7.47, 7.31,
5.00, 3.19, 4.01, 3.07 3.84, 3.42 341 calc: 66.33, 7.70, 7.43 7.19
3.02, 2.63, 1.85, 1.66, found: 66.75, 7.69, 7.36 2.34 1.23
.sup.2D.sub.2O, unless otherwise specified .sup.aDMSO-d.sub.d
TABLE-US-00013 TABLE 9a Analytical Data of Compounds of the
Invention shown in Table 5a ##STR00016## NMR (D.sub.2O) MS elem.
anal. # aryl indan propargyl R3, R4 IR (MH.sup.+) (C, H, N, S) 46
7.48, 7.29, 5.02, 3.19, 4.0, 3.07 3.46, 3.41 calc: 57.97, 6.11,
9.01, 10.30 7.16 3.05, 2.67, found: 58.07, 6.06, 8.85, 10.23 2.37
47 7.50, 7.31 5.04, 3.21, 4.20, 3.09 3.95, 3.87 calc: 59.16, 6.47,
8.62, 9.86 7.19 3.07, 2.70, 3.45, 3.38 found: 59.23, 6.39, 8.52,
9.76 2.38 1.35, 1.32
TABLE-US-00014 TABLE 10 Analytical Data of Compounds of the
Invention shown in Table 6 ##STR00017## NMR.sup.4 MS elem. anal. #
aryl cyclohex. R1 proparg. R3, R4 IR (MH.sup.+) (C, H, N) 40 calc:
found: 41 7.22, 6.95 3.79, 3.26 4.06, 3.01 3.50, 3.36 3227, 2938
287 calc: 63.25, 7.18, 8.68 2.95, 2.32 3.09, 2.96 2768, 1718 found:
63.16, 6.93, 8.69 1.91 1.24, 1.16 1587, 1494 1394, 1301 42 7.21,
7.03 4.60, 2.81 3.88, 3.95 3.01, 2.87 3234, 2936 273 calc: 62.23,
6.80, 9.07 2.72, 2.15 2774, 2130 found: 62.20, 7.01, 9.3 2.02, 1.84
1732, 1499 1.80 1390 43 7.32, 7.12 4.65, 2.88 3.99, 3.04 3.51, 3.37
3216, 2933 287 calc: 63.06, 7.41, 8.65 2.80, 2.20 3.10, 2.96 2768,
2663 found: 63.2, 7.14, 8.81 2.12, 1.94 1.23, 1.16 2129, 1723 1.85
1425, 1399 .sup.4D.sub.2O, unless specified otherwise
BIOLOGICAL EXAMPLES
Example 1
Acetylcholinesterase Inhibition in Mice
1.1 In vitro measurement of Acetylcholinesterase (AChE)
Inhibition
Human erythrocyte acetylcholinesterase (type XIII, Sigma Israel),
was prepared in a stock solution of 1 U/ml, containing Triton (1%)
and bovine serum albumin (0.05%) in phosphate buffer (pH 8). The
enzyme (0.05U) was incubated with 3-5 different concentrations of
test compound (in triplicate) for periods of from 15 to 60 minutes
at 37.degree. C. The substrate acetylthiocholine (0.075M) and
5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB, 0.01M) were then added
and the rate of hydrolysis of the substrate which yields a yellow
product monitored spectrophotomerically at 412 nM (Ellman et al.,
Biochem Pharmacol. (1961) 7: 88-95). The percentage inhibition of
AChE by each concentration of drug is calculated by comparison with
that of enzyme in the absence of drug. The concentration of each
drug that inhibits AChE by 50% (IC.sub.50) at the time of peak
activity was calculated and is given in Table 11 below.
1.2 Ex vivo measurement of Acetylcholinesterase (AChE)
Inhibition
Test drugs or saline were administered sub-cutaneously to male mice
(Sabra strain, 28-35 g). At least 4-5 mice were used per dose and a
minimum of 3 doses per drug were tested. The mice were sacrificed
15, 30, 50, 70, 90, 120 or 180 minutes after drug administration,
the brains rapidly removed (minus cerebellum), weighed and
homogenized in 0.1 M phosphate buffer, pH 8.0, containing Triton (1
mg/100 g tissue) and centrifuged to remove cell debris. Aliquots
(25 .mu.l) of the supernatant were then incubated with
acetylthiocholine and DTNB. AChE activity was measured as described
above. The % inhibition of whole brain AChE by each dose of drug
was calculated by comparison with enzyme activity from 3 saline
treated control mice run at the same time. The dose of each drug
that inhibits AChE by 50% at the peak of activity (ED.sub.50) was
calculated and is given in Table 11.
1.3 Acute Toxicity in Mice
Drugs were administered sub-cutaneously in at least 3 doses, to a
minimum of 10 mice per dose. The dose that was lethal to 50% of the
mice (LD.sub.50) within 6 hours after administration was calculated
for each drug and is given in Table 11. Therapeutic Ratio was
calculated as LD.sub.50 divided by ED50 of ex vivo acetylcholine
esterase inhibition.
Example 2
2.1 Inhibition of MAC activity in vitro
The MAO enzyme source was a homogenate of rat brain in 0.3M
sucrose, which was centrifuged at 600 g for 15 minutes. The
supernatant was diluted appropriately in 0.05M phosphate buffer,
and pre-incubated with serial dilutions of test compounds for 20
minutes at 37.degree. C. .sup.14C-Labeled substrates
(2-phenylethylamine, hereinafter PEA; 5-hydroxytryptamine,
hereinafter 5-HT) were then added, and the incubation continued for
a further 20 minutes (PEA), or 30-45 minutes (5-HT). Substrate
concentrations used were 50 .mu.M (PEA) and 1 mM (5-HT). In the
case of PEA, enzyme concentration was chosen so that not more than
10% of the substrate was metabolized during the course of the
reaction. Deaminated products were extracted into toluene-ethyl
acetate (1:1 v/v) containing 0.6% (w/v) 2,5-diphenyloxazole (ppo)
prior to determination by liquid scintillation counting.
Radioactivity n the eluate indicates the production of neutral and
acidic metabolites formed as a result of MAO activity. Activity of
MAO in the sample was expressed as a percentage of control activity
in the absence of inhibitors after subtraction of appropriate blank
values. The activity determined using PEA as substrate is referred
to as MAO-B, and that determined using 5-HT as MAO-A.
Concentrations of inhibitor producing 50% inhibition of substrate
metabolism (IC.sub.50) were calculated from the inhibition curves,
and are shown in Table 11.
2.2 Inhibition of MAO activity ex vivo
Male Sabra mice, weighing 45-50 g were injected with test compound
solutions (prepared in 0.9% saline). Each dose was administered to
two or three mice. The mice were sacrificed two hours after drug
administration or at a time corresponding to the peak AChE
inhibition time (see Table 11). The brain and liver were rapidly
dissected and stored in appropriate vials on ice. The tissues were
weighed, diluted to 1/2 in sucrose 0.3M and stored at -20.degree.
C. before performance of the MAO assay described above. The results
given in Table 11 relate to measurements made on brain tissue
only.
2.3 Inhibition of MAO activity following sub-acute administration
to rats
Experiments were done in Sprague Dawley male rats. Procedures were
repeated as described in Examples 2.1 and 2.2 but drug
administration was continued daily for 14 days. At the end of this
period animals were sacrificed and MAO levels determined in the
brain, liver and intestines. Compounds 24, 25, 37 and 39 were
administered subcutaneously and/or pet os at a dose of 6 mg/kg(sc)
and 10 mg/kg(po) (compound 24), 25 and 50 mg/kg (compound 25), 45
mg/kg (compound 37) and 40 mg/kg (compound 39). The results are
shown in Table 11a from which it can be seen that these compounds
displayed selectivity in inhibiting MAO enzyme sub-types in the
brain in preference to the periphery.
TABLE-US-00015 TABLE 11 AChE Inhibition Time Ex vivo to return to
MAO-B Inhibition MAO-A Inhibition Acute Toxicity ED50 to peak 50%
of Ex vivo Ex vivo LD50 Therapeutic In vitro .mu.moles/kg activity
peak In vitro ED50 In vitro ED50 .mu.moles/kg Ratio # IC50 .mu.m
(AC) t (min) t (min) IC50 .mu.m .mu.moles/kg IC50 .mu.m
.mu.moles/kg (LD) LD/AC 1 0.6 5.0 30 >120 >1000 >>80 75
>>80 83.8 16.8 23 3.5 22.4 15 70 600 100 800 >120 255 11.4
2 7.3 NT >1000 32 3 20.0 46.3 60-90 >180 >1000 12.6 950
20.6 25 53.0 140.0 60 >180 >1000 200 270 >>350 1400
10.8 26 17.0 120 30-60 264 333 114 >>440 1200 9 27 5.72 30 15
>60 >1000 >>160 >1000 >>160 300 10 28 100.0 NT
5 11.5 85.0 60 >120 >>277 >>277 840 9.9 7 32.0 NT
>1000 600 8 1.0 10.0 15-30 >60 >1000 >>50 50
>>50 87 8.7 9 0.18 19 15 93 4.9 29 8.5 53.7 15 >60 40 30
40 50 500 9.3 10 38.0 34.7 60-90 >180 >1000 >175 22
>175 740 21.3 30 1300.0 NT 31 10.0 110 >1000 >100 >1000
>100 32 3.7 7.8 15 500 >>20 190 >>20 68 9.0 12 2.0
8.0 15 >1000 130 <20 <2.5 33 540.0 NT >1000 1000
>1000 >>1200 34 0.046 0.65 30 100 0.5 3.7 5.7 35 2.2 10 60
100 <1 33 3.3 37 51 125 500 200 750 >200 1700 13.6 39 36 80
30-60 >180 1000 >>200 550 >>200 1150 14.4 24 3 16.6
15 750 100 850 >120 179 10.8 60 42 58 51 >1000 300 54 1.8
>100 >100 55 2 >100 >100 56 11.5 180 57 2.4 70 25 89 48
10 49 2 17 4 16 9 50 0.26 61 0.75 47 500 >100 700 >100 64 1.9
13.2 >1000 >120 1000 >120 150 11.4 38 33 >1000 10
>400 170 >400 36 15 >400 >1000 >100 >1000 >100
>1000 62 0.57 290 60 100 >>200 80 >>200 63 2.5 140
60-90 120 >300 40 >300 1300 9.3 71 29 >100 130 >100 72
38 >200 >100 >100 78 10 101 60-90 >120 450 >>450
1300 12.9 79 9.4 94 90 >180 >>450 >>450 1000 10.6 81
11.5 40 90 >120 >>100 >>100 920 23 83 80 86 10.5 87
9.1 85 17 >100
TABLE-US-00016 TABLE 11a Effect of Compounds 24, 25, 37 and 39 on
MAO activity after chronic sub-acutetreatment to rats % MAO-A
inhibition 24 % MAO-B inhibition Compound 6 (sc) 25 37 39 24 25 37
39 Dose (mg/kg) 10 (po) 25 50 45 40 25 25 50 45 40 Brain sc 30 53
75 78 17 50 61 85 87 27 po 0 70 67 20 80 82 Intestine sc 0 0 30 0 0
0 29 45 26 40 po 30 25 0 20 30 21 Liver sc 0 0 10 0 0 0 14 40 29 0
po 10 25 28 0 35 28
Example 3
Effect of Drug Treatment Following Closed Head Injury (CHI) in
Mice
The procedure for closed head injury followed was as described for
rats in Shohami, et al. (J Neurotrauma (1993) 10 (2): 109-119) with
changes as described.
Animals: Male Sabra mice (Hebrew University strain) weighing 34-40
g were used. They were housed in groups of 10 per cage, in a 12
hr:12 hr light:dark cycle. Food and water were provided ad
libitium.
Trauma was induced under ether anesthesia. A longitudinal incision
was performed in the skin covering the skull and the skin retracted
to expose the skull. The head was fixed manually at the lower plane
of the impact apparatus. A weight of 333 g was delivered by an
electric device from a distance of 3 cm to the left hemisphere, 1-2
mm lateral to the midline in the midcoronal plane. Test compounds
were injected sub-cutaneously at a dosage corresponding to the
ED.sub.50 acetylcholinesterase, once 15 min. after CHI.
3.1 Assessment of Motor Function
Motor function and reflexes were evaluated in the injured mice at
different times after closed head injury (CHI) using a neurological
severity score (NSS) as shown in Table 12 below, which is modified
from that described for rats (Shohami, et al. supra.). One point
was awarded for the lack of a tested reflex or for the inability to
perform the tasks outline in the Table. The maximal score that can
be reached at 1 hour post-CHI is 25 points and 21 at later times.
The difference in NSS at 1 hr and at any other time reflects the
recovery, and is referred to as .DELTA.NSS. An NSS score of 15-19
at 1 hr denotes severe injury, 11-14 moderate injury and less than
10 mild injury. The NSS recorded after treatment with test compound
or control is shown in Table 13.
TABLE-US-00017 TABLE 12 Neurological Severity Score for mice after
Closed Head Injury Points at Points at any Parameter 1 hour other
time Inability to exit from a circle (30 cm diameter when left in
its center for 30 min 1 for 60 min 1 for >60 min 1 1 Loss of
righting reflex for 10 second 1 for 20 seconds 1 for >30 seconds
1 1 Hemiplegia - inability of mouse to 1 1 resist forced changes in
position Flexion of hind limb when 1 1 lifted by tail Inability to
walk straight when 1 1 placed on the floor Reflexes Pinna reflex 1
1 Corneal reflex 1 1 Startle reflex 1 1 Clinical grade Loss of
seeking behaviour 1 1 Prostration 1 1 Loss of reflexes Left
forelimb 1 1 Right forelimb 1 Left hindlimb 1 1 Right hindlimb 1 1
Functional test Failure in beam balancing task 1 1 (0.5 cm wide)
for 20 seconds 1 1 for 40 seconds 1 1 for >60 seconds Failure in
round stick balancing task (0.5 cm is diameter for 10 seconds 1 1
Failure in beam walking task 3 cm wide 1 1 2 cm wide 1 1 1 cm wide
1 1 Maximum Points 25 21
TABLE-US-00018 TABLE 13 Change in Neurological Severity Score after
Closed Head Injury in Mice .DELTA.NSS, 24 hr .DELTA.NSS, 7 days
.DELTA.NSS, 14 days Drug/dose N post-CHl post-CHl post-CHl Saline,
1 ml/kg 51 4.75 .+-. 0.17 5.83 .+-. 0.36 5.96 .+-. 0.4 1 (1.3
mg/kg) 10 5.50 .+-. 0.34.sup.m 7.31 .+-. 0.45.sup.m 9.21 .+-. 0.47
24 (6.5 mg/kg) 12 6.31 .+-. 0.23.sup.m 8.67 .+-. 0.41.sup.m 9.67
.+-. 0.66.sup.m 25 (46 mg/kg) 10 5.00 .+-. 0.42 7.42 .+-.
0.62.sup.m 9.01 .+-. 0.69.sup.m 25.sup.1 (46 mg/kg) 10 4.90 .+-.
0.43 7.70 .+-. 0.33.sup.m 8.80 .+-. 0.33.sup.m 10 (15 mg/kg) 11
5.36 .+-. 0.39 6.64 .+-. 0.41.sup.m 6.73 .+-. 0.52 37 (30 mg/kg) 12
5.50 .+-. 0.26 6.92 .+-. 0.38 8.25 .+-. 0.62 39 (30 mg/kg) 14 5.36
.+-. 0.25 6.71 .+-. 0.45 7.64 .+-. 0.48 .sup.1administered 60 min
before CHl .sup.msignificantly different from saline control (p
< 0.05)
3.2 Assessment of Reference Memory
Morris Water Maze Test: the water maze consists of a circular
aluminium pool, im in diameter and 60 cm in depth, filled with
water to a depth of 17.5 cm. The hidden goal platform is a glass
vessel (15 cm diameter.times.16.5 cm height) placed upside down at
a fixed location in the pool, 1 cm below the surface of the water.
The water temperature is maintained at 24.degree. C. and the pool
is always placed in the same position in the room to provide the
same extra-maze cues. Prior to CHI (as described in Example 3
above), mice were given 3 trials per day for 5 consecutive days to
establish a baseline performance--measured as the latency to find
the platform from the same start location. Commencing 24 hr after
CHI, mice were retested daily for 2 weeks in 3 trials per day.
FIGS. 1, 2 and 3 show the reduction in latency for mice treated
with compounds 24 (6.5 mg/kg), 25 (46 mg/kg), 1 (1.3 mg/kg), 10 (15
mg/kg), 37 (30 mg/kg) or 39 (30 mg/kg) compared to saline treated
controls after CHI. It appears that immediately post-CHI mice
forget the location of the goal. Memory is enhanced following
treatment with test compounds, as compared to saline treated mice.
In the Figures the arrow shows the time of CHI.
Example 4
Effect On Mice Having Experienced A Hypobaric Hypoxic Episode
The hypobaric hypoxic model is a well accepted model for assessing
the activity of compounds believed to possess neuroprotective
activity. The model is based on that described in Nakanishi, M., et
al. Life Sci. (1973) 13: 467, Oshiro, et al., J. Med. Chem. (1991)
34: 2004-2013 and U.S. Pat. No. 4,788,130.
A 12 liter desiccator (desiccator A) and a 2.5 liter desiccator
(desiccator B) were separately connected to a vacuum pump.
Desiccator B was disconnected and allowed to equilibrate with room
air whilst desiccator A was evacuated to a pressure of 100 mmHg.
Four male ICR albino mice (22-28 g) were placed in desiccator B.
Desiccator B was then closed to room air and connected to
desiccator A. The pressure inside desiccator B was monitored using
a mercury manometer and at the point were the pressure in
desiccator B reached 200 mmpg (usually within 14 seconds), the two
desiccators were disconnected from the vacuum pump and the pump
switched off. The survival time from the moment of induction of
hypoxia to the time of cessation of respiration was recorded for
each mouse for a maximum of 15 minutes after which time room air
was reintroduced to desiccator B. Survivors were monitored for
signs of lethargy or vitality.
Effect of drug treatment was assessed as the percent of the
survival time of the drug treated group with respect to the saline
injected or vehicle injected control group. Control groups were run
twice, before and after each experimental group and consisted of 8
mice in groups of 4 mice to ensure a constant residual volume of
oxygen in all tests. The effect of each dose of test drug was
determined in duplicate i.e. two groups of 4 mice. The range of
survival times of control mice was from 108-180 seconds.
Positive reference drugs were sodium pentobarbital at a dose of 40
mg/kg, and diazepam 10 mg/kg given 0.5 h prior to hypoxia,
physostigmine 0.2 and 0.4 mg/kg and neostigmine 0.2 mg/kg given sc
30 min before hypoxia. Methyl atropine 1 mg/kg was given sc. 10
min. before physostigmine.
Test drugs were dissolved in 0.9% saline, and injected sc. in the
nip of the neck at a dose in accordance with body weight, 60-90
min. before hypoxia. The volume of injection was 0.2-0.3 mL per
mouse (10 mL/kg). The initial dose was about one third of the
reported LD.sub.50 for acetylcholine esterase inhibition. If no
protection could be obtained, the dose was further increased to the
nearest non-toxic dose. In case of protection, the dose was further
reduced in an attempt to locate the "protective" dose range.
Per cent survival times as compared to saline treated control is
shown in Table 14.
TABLE-US-00019 TABLE 14 Survival Time of Mice Having Experienced a
Hypobaric Episode Time of dose Dose (min before Protection Compound
mg/kg hypoxin) (% of control) p Control 100 (saline) Nembutal 40 30
253 .+-. 200 <0.005 Diazepam 10 30 316 .+-. 78 <0.003
Neostigmine 0.2 30 141 .+-. 32 <0.01 Physostigmine 0.2 30 453
.+-. 222 <0.001 0.4 30 552 .+-. 210 <0.001 Physostigmine 0.4
30 296 .+-. 193 <0.05 and Atropine 1.0 40 methyl nitrate 1 8 60
637 .+-. 116 0.007 4 60 470 .+-. 200 0.001 2 60 120 .+-. 51 NS 24
50 60 738 .+-. 00 <0.001 21 60 269 .+-. 166 <0.02 25 100 60
761 .+-. 91 0.001 75 60 559 .+-. 225 0.001 50 60 380 .+-. 231 0.01
25 60 84 .+-. 35 NS 27 50 60 455 .+-. 23 <0.001 3 60 287 .+-.
319 <0.001 15 60 143 .+-. 56 <0.05 8 60 119 .+-. 45 NS 29 77
60 508 .+-. 206 <0.001 53 60 638 .+-. 10 <0.001 25 60 131
.+-. 56 NS 25 30 273 .+-. 183 <0.02 10 50 90 705 .+-. 101 0.001
25 90 700 .+-. 201 0.001 10 90 304 .+-. 129 0.001 12 20 60 725 .+-.
128 <0.001 15 60 649 .+-. 221 <0.001 10 60 386 .+-. 238
<0.01 7 60 248 .+-. 97 <0.001
Example 5
Neurological Score and Brain Infarct Size in Male Wistar Rats After
Middle Cerebral Artery Occlusion (MCA-O)
A modification of the procedure described by Tamura, et al was used
(Tamura A, Graham D1, McCulloch J, Teasdale G H (1981) J. Cereb.
Blood Flow and Metab. 1: 53-60). Male Wistar rats (Olac
England-Jerusalem) 300-400 g each were anesthetized with a solution
of Equitesine administered i.p. at a dose of 3 ml/kg. Equitesine
consists of 13.5 ml sodium pentothal solution (60 mg/ml), 3.5 g
chloral hydrate, 1.75 g MgSO.sub.4, 33 ml propylene glycol, 8.3 ml
absolute alcohol, made up to 83 ml with distilled water.
Surgery was performed with the use of a high magnification
operating microscope, model SMZ-2B, type 102 (Nikon, Japan) In
order to expose the left middle cerebral artery, a cut was made in
the temporal muscle. The tip of the coronoid process of mandible
was excised as well and removed with a fine rongeur. Craniectomy
was made with a dental drill at the junction between the median
wall and the roof of the inferotemporal fossa.
The dura matter was opened carefully using a 27 gauge needle The
MCA was permanently occluded by microbipolar coagulation at low
power setting, beginning 2-3 mm medial to the olfactory tract
between its cortical branch to the rhinal cortex and the laterate
striate arteries. After coagulation, the MCA was severed with
microscissors and divided to ensure complete occlusion. Following
this, the temporalis muscle was sutured and laid over the
craniectomy site. The skin was closed with a running 3-0 silk
suture. A sham craniectomy operation was performed on a parallel
group of rats, but without cauterization of the MCA.
During the entire surgical operation (20-25 min) in either group,
body temperature was maintained at 37 to 38.degree. C. by means of
a body-temperature regulator (Kyoristsu, Japan) consisting of a
self-regulating heating pad connected to a rectal thermistor. At 24
and 48 hours post surgery a neurological score was taken in order
to assess the severity of the injury in the drug-treated rats with
respect to their untreated controls.
Drugs were administered as an s.c. injection, according to the
following schedule:
Compound 24: 7.8 mg/kg 15 minutes prior to MCA-O and 6.5 mg/kg 2
hours post MCA-O.
Compound 25: 43 mg/kg 90 minutes prior to MCA-O and 30 mg/kg 3
hours post MCA-O.
After 48 hours of ischemia induced by permanent occlusion
morphometric, the animals-were anesthetized with Equitesine and
measurement of infarct volume was performed as follows by TTC
(2,3,5-triphenyl tetrazolium chloride) staining. TTC 1% in saline
was prepared immediately before use and protected from exposure to
light by aluminum foil wrap. MCA-O rats were deeply anesthetized
and a 23-gauge butterfly needle with an extended tubing and a 20 ml
syringe was inserted into the ventricle via thoracotomy. The right
atrium was incised to allow outflow of saline. Heparine 50 i.u. in
saline was delivered until the perfusate was bloodless. A 30-ml
TTC-filled syringe was exchanged for the saline syringe and TTC was
injected into the left ventricle at a rate of 5 ml/min. Both
perfusate solutions were administered at 37.5.degree. C. The brains
were removed and immersed into 20 ml of 1% TTC contained in tightly
closed glass vials. These were further placed for 2 hours in a
water bath maintained at 37.degree. C. The TTC solution was
decanted, the brains removed, wiped dry and placed into 10%
buffered formalin solution for 3 days. Six coronal slices each 2 mm
thick, 3, 5, 7, 9, 11 and 13 mm distal from the frontal pole were
obtained with a brain matrix (Harvard Apparatus, South Natick,
Mass.). Infarction areas were measured with a video imaging and
analyzer from both sides of the coronal slices and expressed in
mm.sup.2. The volume of the infarcted region in mm was calculated
by taking the sum of the ischemic areas in all six slices. The
volume of infarcted region for the saline control and compounds 24
or 25 are given in Table 15a.
Neurological Score
The neurological score was measured in a manner slightly different
from that given in Example 3. This method consists of the sum total
of a series of ratings assigned to the performance of specific
locomotor activities in a given rat. The scale runs from 0 (fully
normal rats) to 13 (fully incapacitated rats). Most parameters are
rated as either 0 (normal), or 1 (incapacitated) others are graded.
The following tests were used in the present study:
General observation tests: hypoactivity, sedation,
piloerection.
Motor reflex. Rats were lifted by the tail about 15 cm above the
floor. Normal rats assume a posture in which they extend both
forelimbs towards the floor and spread that hind limbs to the sides
in a trapeze-like manner. MCAO, when severe, causes consistent
flexion of the contralateral limb.
Motor ability. This is seen as the ability to grasp a rod 1 cm in
diameter by the contralateral limb for 5-15 sec when the rat is
left hanging on the rod through the arm pit.
Motor coordination. Normal rats are able to walk up and down a
beam, 5 cm wide placed at a moderate slant. Failure to walk the
beam in either direction reveals some motor incoordination, lack of
balance and limb weakness.
Gait. Ability to restore normal position to either hand
contralateral or fore contralateral limb when intentionally
displaced while on a narrow beam.
Balance. Ability to grasp and balance on a narrow beam 2 cm
wide.
Locomotor activity. Total movements over a period of 15 min in an
automated activity cage.
Ratings assigned to each of the above parameters are given in Table
15.
TABLE-US-00020 TABLE 15 Neurological scores assigned to each of 10
parameters of posture and locomotion Parameter Score a. Activity in
home cage normal = 0 hypoactive = 1 b. Sedation none = 0 marked = 1
c. Piloerection none = 0 marked = 1 d. Extenstion of contra- good =
0 flexed limb = 1 lateral forelimb to- wards floor when lifted by
tail e. Spread of contralateral good = 0 flexed limb = 1 hind limb
when lifted by tails (trapezoid posture) f. Grasp rod with contra-
good = 0 poor = 1 lateral limb for 5-15 sec. when suspended by
armpit g. Walk on beam 5 cm wide good = 0 poor = 1 h. Restoration
of contra- good = 0 poor = lateral hind and/or 1 (one limb)
forelimb to original 2 (two limbs) position when intentionally
displaced i. Grasping & balance on good = 0 poor = 1 beam 2 cm
wide j. Motor activity with 0-25% of control = 3 respect to control
26-50% of control = 2 (15 min in activity cage) 51-75% of control =
1 76-100% of control = 0 k. Tendency to lean on 1 contralateral
side l. Contralateral circling 1 when pulled by tail m.
Contralateral circling 1 spontaneous.
Table 15a shows the effect of compounds 24 and 25 in this model,
comparing the change in NSS measured in 24 and 48 hours post
injury.
TABLE-US-00021 TABLE 15a Volume infarction Compound .DELTA.NSS*
Mean .+-. SD mm* Saline 0.745 211 .+-. 75 24 1.625 152 .+-. 45 25
1.78 189 .+-. 54 *Difference is .DELTA.NSS measured at 24 hours and
48 hours. From this it can be seen that compounds 24 and 25 have a
longer lasting effect than the saline treated control.
##STR00018##
* * * * *