CA2146000A1 - 3,3-disubstituted tri-and tetracyclic indolin-2-ones useful for the treatment of cognitive disorders - Google Patents

Abstract

Compounds of formula (I) have been shown to enhance the release of the neurotransmitter acetylcholine, and thus may be useful as chemical intermediates and as pharmacological agents in the treatment of diseases of man, such as in Alheimer's Dis-ease and other conditions involving learning and cognition, where subnormal levels of this neurochemical are found. The com-pounds of this invention have the structure shown in formula (I).

Description

DM-6617 2 1 ~ 6 0 0 G PCJ~ 9 3 / C . . ~ tZ ~

3,3-Disubstituted Tri- and Tetracyclic Indolin-2-ones Useful for the Treatment of Cognitive Disorders.
Fi~l ~ of ~h~ ~n~n~ n This invention relates to 3,3-disubstituted tri-and tetracyclic indolin-2-ones, to pharmaceutical compositions thereof, and methods of use in mammals to treat cognitive disorders, neurological dysfunction, and/or mood disturbances such as, but not limited to 10 degenerative nervous system diseases. Additionally, these compounds can be used as reagents in studies on the biochemical mechanism of neurotransmitter diseases.
~ . o,..~l In~l n~i n~ Pr; ~r l~rt Increasingly there is a need for effective treatments for nervous systems disorders and neurological deficiencies. Many of these diseases correlate with increasing age due mainly to degenerative changes in the nervous system. Although in early stages of some diseases, certain systems are rather specifically affected (e.g., cholinergic systems in Alzheimer's Disease and Myasthenia Gravis, the dopaminergic system in Parkinson's Disease, etc.) multiple neurotransmitter systems deficiencies (acetylcholine, dopamine, norepinephrine, serotonin) are generally found at later stages of diseases such as senile dementia, multi-infarct dementia, Huntington's Disease, mental retardation, etc. This explains the generally observed multiple symptomology that includes cognitive, neurological and effective/psychotic components (see Gottfries, PsychoDharmacol., 86, 245 (1985)). Deficits in the synthesis and release of acetylcholine in the brain are generally thought to be related to cognitive impairment (see Francis, et al., New Fngl~n~ J. Med., 7, 313 (1985)) whereas neurological deficits (e.g. Parkinsonian symptoms) and mood/mental changes may be related to impairment of dopaminergic and serotonergic systems, respectively. Other neurological ~ DM-6617 21 ~ ~ ~ Q ~ R C ~ r ~

deficits (e.g., Myasthenia Gravis) are related to cholinergic deficiencies in the peripheral nervous system.
Treatment strategies employed previously encompass 5 vasoactive drugs like vincamine and pentoxifylline;
metabolic enhancers like ergoloid mesylates, piracetam, and naftidrofuryl; neurotransmitter precursors like L-DOPA, choline, 5-hydroxytryptamine; transmitter metabolizing enzyme inhibitors such as physostigmine;
10 neuropeptides like adrenocorticotropic hormone and ~~
vasopressin-related peptides. Except for L-DOPA
treatment for Parkinson's Disease and cholinesterase inhibitor treatment for Myasthenia Gravis, these treatment strategies have generally failed to enhance 15 the residual function of the affected systems by enhancing the stimulus-induced release of -=
neurotransmitters. Theoretically, such an enhancement-would improve the signal-to-noise ratio during chemical transmission for information, thereby reducing deficits in processes related to cogn tion, neurological function and mood regulation.
~ uropean Patent Application 311,010 discloses that a,a-disubstituted aromatics or heteroaromatics of the formula:

r x ~ ~CH2-Het 1 ~ y~ ~CH2-Het 2 or a salt thereof, are useful as cognition enhancers.
U.S. Patent No. 4,760,083 to Myers, et al.
discloses that indolines of the following formula are useful for treatment of cognitive deficiencies:

~ ~M-6617 2 1 4 6 0 0 D r ~ J ~

x ~=H2~) Y (CH2)p The references claim the necessity of two heteroaryl groups for activity.
European Patent Application No. 0 415 102 Al by Effland, et al. describes an invention related to the formula:

\ R
(Y) n ~=0 ~X) m U.S. Patent No. 3,S9S,866 to D. E. Butler describes an invention of the formula:

~ A NR1R2 ~0 y - Ar ~uropean ~atent Application No. 0 347 698 Al wherein Ting, et al. describes a compound of formula:

DM--6617 21 ~ ~ 0 0 ~ ~ g~ . ~ ~ ~ U , 7 y~R 3 ~X

Patent WO 91/01/306 1991 discloses oxindole derivatives of formula:
-R2~ R3 R1 ~Xo o~~ R~' useful for treating senile dementia, i. e. improving brain functions and activating and protecting brain 10 metabolism. This reference only discloses imides and does not suggest alkyl or aryl substituted amides.
.~?~ of 1~h~ TnV~n~ n Presently it has been found that certain 3,3-disubstituted tri- and tetracyclic indolin-2-ones enhance the stimulus-induced release of neurotrans-mitters, specifically acetylcholine in nervous tissue, and thus improve processes involved in learning and memorization of an avoidance task.
According to the present invention, there are provided compounds of formula:

2l46a~ 93/f~7 DM- 6 6 1 7 ~ . R O ~

R5--~o X~J~ R3 I

or pharmaceutically acceptable salts thereof, wherein:

X is a single bond, O, S, SO, SO2, CH2, CH2CH2, CH=CH, C=O, C(R7)(oR6), CH(OR6), -CONR6-, -NR6CO-, -CH2-NR6-, -NR6-CH2-, NR6, -C(R7)=N-, -CH(R7)-N(R6)-, or -CR7-, -CH- when a iq a single bond and b is a double bond;

a and b are each single or double bonds, provided that a is a single bond when b is a double bond, and that b is a single bond when a is double bond, and that b is a double bond when X is -CH- or -CR~- and when a is a single bond;

R1 is 2-, 3- or 4-pyridyl, or 4-pyrimidinyl;
R2 is ~(CH2)m-W, wherein m = 1-4, and W is selected from the group:
(a) 2-, 3- or 4-pyridyl, 2s (b) 2-, 4-, or 5-pyrimidinyl, (c) 2-pyrazinyl, (d) 3- or 4-pyridazinyl, (e) 3- or 4-pyrazolyl, (f) 2- or 3-furyl, (g) 2- or 3-tetrahydrofuranyl, (h) 2- or 3-thienyl, DM-6617 - 21 160D~ RC - -(i) 3-indolyl, (j) aryl unsubstituted or substituted with 1-3 R5, (k) 2-fluoro-4-pyridyl, or s R2 is - (CH2) n~Y~
wherein n = 1-6, and Y is selected from the group:
-CH=CHCo2R7, -CH=CHCoR7, -CH=CHR7, -CH=C(R7)2, -CH=CH2, -C_CCo2R7, -C_CCOR7, -C_CR7, or -C_CH, F, Cl, Br, oR6, N(R6)2, CO2H, Co2R7, CoNHR7, NHCHO, CONHR6, CON(R7)2, CN, -oCoR7, CoR7, CHO, SR7, SoR7, So2R7 or NO2;

R3 and R4 are each independently selected from the group:
H, alkyl of 1-6 carbons, alkenyl of 2-6 carbons, alkynyl of 2-6 carbonsi cycloalkyl of 3-7 carbons, cycloalkylalkyl of 3-10 carbons, aryl unsubstituted or substituted with 1-3 R5, alkaryl of 1-10 carbons, provided that when a is a single bond R3 is =O, =CH2, =CH(R7), =C(R7)2, or (CH3)2, and provided that when a and b are each a single bond, then R3 and R4 are each independently selected from the group: =O, =CH2, =CH(R7), =C(R7)2, (CH3)2 i or R3 a~d R4, taken together may form a saturated or unsaturated carbocyclic or heterocyclic ring, unsubstituted or substituted with 1-2 R5 substituents;

R5 is selected from the group:
H, alkyl of 1-6 carbons, aryl unsubstituted or substituted with 1-3 R8, alkaryl of 1-10 carbons, F, Cl, Br, I, oR6, NHR6, N(R6)2, CO2H, CO2R7, CoNHR7, CON(R7)2, CN, CoR7, CHO, SR7, SoR7, SO2R7, NO2 or -CH=CH-CH=CH- which ~ ~- 93/ C t7 DM-6617 21~6~ ~ O ~ ~ ~ rL I ~ 3 ~ 4-~ attaches to an adjacent carbon atom forming a fused ring;

R6 is independently selected at each occurrence from the group:
H, alkyl of 1-6 carbons, aryl unsubstituted or substituted with 1-3 R8, alkaryl of 1-10 carbons, -So2-R7~ and -CoR7;

R7 is independently selected at each occurrence from the group:
alkyl of 1-6 carbons, aryl unsubstituted or substituted with 1-3 R8, and alkaryl of 1-10 carbons;

R8 is independently selected at each occurrence from the group:
OR9, NHR9, N(R9)2, CO2H, CO2R9, CONHR9, CON(R9)2, CN, CoR9, CHO, SR9, SoR9, SO2R9, and NO2; and R9 is independently selected at each occurrence from the group:
H, alkyl of 1-6 carbons and aryl.
Pr~farr~ zm~m~
Preferred compounds of this invention are those of Formula I wherein, together or independently:

X is a single bond, O, S, SO, SO2, CH2~ CH2CH2, CH=CH, C=O, NR6;

a is a single or a double bond;

b is a single bond;
R1 is 2-, 3- or 4-pyr1dyl, or 4-pyrimidinyl R2 is ~(CH2)m-W, DM-6617 2 1 ~ ~ O O O ~ ~

wherein m = 1-4, and W is selected from the group:

2-, 3- or 4-pyridyl, 2-, 4-, or 5-pyrimidinyl, or 2-pyrazinyl; or R2 is ~(CH2)n~Y, wherein n is 1 to 6, and Y is selected from the group:
Co2R7, CN, CoR7, CHO, -OCOR7;

R3 and R4 are each independently selected from the group:
H, alkyl of 1-6 carbon atoms or -CH=CH-CH=CH-to form a fused ring; and R5 is selected from the group:
H, alkyl of 1-6 carbons, phenyl unsubstituted or substituted with 1-3 R8, F, Cl, Br, I, NO2 or -CH=CH-CH=CH- which attaches to an adjacent carbon forming a fused ring.
More preferred compounds of this invention are those preferred compounds wherein:

X is O, S, SO, SO2, CH2, CH2cH2~ C=Oi R2 is ~(CH2)m-W, wherein m = 1, and W is selected from the group:
2-, 3- or 4-pyridyl, and 4-pyrimidinyl; or R2 i s - ( CH2 ) n~Y , where n = 3-4, and Y is selected from the group:
C32R7, CN, and -CCoR7;

R3 and R4 are each H, or R3 and R4 join together forming a fused ring consisting of -CH=CH
CH=CH-; and R5 is selected from the group:

~14600~ `~ 93 ' n~ !, 7 DM-6617 RQ, ~S

H, Cl, ~r, I, ~2 or -CH=CH-CH=CH- which attaches to an adjacent carbon atom forming a fused ring.

s Specifically preferred are those compounds of Formula I which are:

(a) 2,2-Bis~4-pyridinylmethyl)-pyrrolo[3,2,1-kl]phenothiazin-1(2H)-one;

(b) 1,1-Bis(4-pyridinylmethyl)-5,6-dihydro-4H-pyrollo[3,2,1-ij]quinolin-2(lH)-one;

(c) 2,2-Bis(4-pyridinylmethyl)-6,7-dihydro-indolo[1,7-ab][l]benzapin-1(2H)-onei .
(d) 2,2-Bis(4-pyridinylmethyl)-pyrrolo[3.2.1-kl]phenoxazin-1(2H)-one;

(e) 2,3-Dihydro-6,6-bis(4-pyridinylmethyl)-pyrrolo[1,2,3-de]-1,4-benzothiazin-5(6H)-one;

(f) 1,2-Dihydro-2-(4-pyridinylmethyl)-2-(pentanenitrile)-pyrrolo E 3.2.1-kl]phenoxazin-1(2H)-one, hydrobromide.

It should be recognized that the above-identified groups of compounds are preferred embodiments of this invention, but that their description herein is in no way intended to limit the overall scope of this invention.
This invention also provides pharmaceutical compositions comprising a suitable pharmaceutical carrier and an amount of one or more of the above-described compounds effective to treat cognitive orneurological dysfunction. Still further, this invention relates to a method of treating cognitive or DM-6617 2146000 ~; - 93 ... ?~
... ..

neurological dysfunction in a mammal comprising administering to the mammal a therapeutically effective amount of one or more of the above-described compounds.
V~ n .~f ~hQ ~nvention s The compounds herein described may have asymmetric centers. All chiral, enantiomeric, diastereomeric, and racemic forms are included in the present invention.
Thus, the compounds of Formula (I) may be provided in the form of an individual stereoisomer, a non-racemic 10 stereoisomer mixture, or a racemic mixture.
Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention.
When any variable occurs more than one time in an;y constituent or in Formula (I), or any other formula .- -herein, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are 20 permissible only if such combinations result in stable compounds.
As used herein, "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon 25 atoms. As used herein "alkoxy" represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge; "cycloalkyl" is intended to include saturated ring groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyli and "biycloalkyl" is intended to include saturated bicyclic ring groups such as [3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane (decalin), [2.2.2]bicyclooctane, and so forth. "Alkenyl" is intended to include hydrocarbon chains of either a straight or 35 branched configuration and one or more unsaturated carbon-carbon bonds which may occur in any stable point along the chain, such as ethenyl, propenyl, and the like; and "alkynyl" is intended to include hydrocarbon chains of 2 1 4 6 ~

- R~

either a straight or branched configuration and one or more triple carbon-carbon bonds which may occur in any stable point along the chain, such as ethynyl, propynyl and the like. "Cycloalkyl-alkyl" is intended to include cycloalkyl attached to alkyl. "Halo" as used herein refers to fluoro, chloro, bromo, and iodo; and "counterion" is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, sulfate, and the like.
As used herein, "aryl" or "aromatic residue" is intended to mean phenyl or naphthyl; "carbocyclic" is intended to mean any stable 5- to 7- membered monocyclic or bicyclic or 7- to 14-membered bicyclic or tricyclic carbon ring, any of which may be saturated, partially 15 unsaturated, or aromatic, for example, indanyl or ~ tetrahydronaphthyl ~tetralin).
As used herein, the term "heterocycle" is intended to mean a stable 5- to 7- membered monocyclic or bicyclic or 7- to 10-membered bicyclic heterocyclic ring which is either saturated or unsaturated, and which consists of carbon atoms and from l to 3 heteroatoms selected from the group consisting of N, O and S and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen may optionally be quaternized, and including any 2s bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. The heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom which results in a stable structure. The heterocyclic rings described herein may be substituted on carbon or on a nitrogen atom if the resulting compound is stable. Examples of such heterocycles include, but are not limited to, pyridyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, benzothiophenyl, indolyl, indolenyl, quinolinyl, isoquinolinyl or benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, DM-6617 21460Q0 ~ ~ 93 / C 4 7 6 `:
.

decahydroquinolinyl, pyrazinyl, quinazzoyl, phthalazinyl, naphthyridinyl or octahydroisoquinolinyl.
The term "substituted", as used herein, means that one or more hydro~en atom(s) on the designated atom is s replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution results in a stable compound.
By "stable compound" or "stable structure" is meant 10 herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
As used herein, "pharmaceutically acceptable salts"
15 refer to derivatives of the disclosed compounds that are-modified by making acid or base salts. Examples include,-but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids.
Pharmaceutically acceptable salts of the compounds of the invention can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous 25 media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Reminston's Pharm~ceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA, 1985, p. 1418, the - disclosure of which is hereby incorporated by reference.
AS used herein, the term "therapeutically effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human subject that is being sought by a clinician or researcher.
S,yr~t- h~.~ i ~
The compounds of the present invention may be prepared according to the following schemes and examples, using appropriate materials and are further - ~ DM-6617 2 1 4 6 0 0 ~ ? " `- ~ s ~
.; ~, ~ , exemplified by the following specific examples. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare s those compounds.
Scheme 1 shows one process for the preparation of compounds of the present invention. In this scheme a compound of formula 2 is reacted in an inert solvent with chloroacetyl chloride to provide a compound of formula 3. The solvent is preferably an aromatic hydrocarbon such as benzene or toluene or a halohydrocarbon such as methylene chloride, 1,2-dichloroethane or chloroform. The reaction temperature is not critical and generally ranges from about 20C to about 120C. Preferably, the reaction temperature is the reflux temperature of the solvent. Next, fusion of a compound of formula 3 with anhydrous aluminum chlor~de at a temperature within a few degrees of the melting point of the compound of formula 3, but using a temperature no greater than about 200C, affords a compound of formula 4. This reaction is disclosed in the following references which are herein incorporated by reference: Chem. Ber. 47, 2120 (1914), J. Chem. Soc.
1697 (1954), Tetr~he-lron 24, 6093 (1968) and J. Med.
Ch-om. 15, 762 (1972). See also, the references listed in Table I. Next, the anion of a compound of formula 4 is generated by treatment with a base, in an appropriate aprotic solvent and at a suitable temperature. The resulting anion is then alkylated with an appropriate alkyl halide ~R1CH2-Hal) to give a compound of formula 5. This process can then be repeated by treatment of a compound of formula 5 with a base, followed by addition of a second alkylating agent (R2-Hal) to produce a compound of Formula I.
3s The temperature and duration of the alkylation reaction are not critical, and may be varied over a wide range from room temperature for 24 hours to 80C for 3 hours. Preferred conditions are room temperature, and a - duration of 2-3 hours. Equivalent amounts of the reagents can be used, but it is preferable to use the haloalkylating in a slight excess This method is disclosed by Myers and Nickolson, in US Patents 4,876,259 and 4,760,083, which are herein incorporated by reference.

DM-6617 214-6000 RO ~ r ~ 9~/ 0.-J4.7A~

~ Sch~me 1 Cl R5--~ H C~OCH2CI R5--~ ~cO AIC13 X~l~ R3 X~l~ R3 R5--~0 R~CH2-Hal Rs_~=O

R~ 4 Rl~ R~ 5 r i- -R2-Hal ~ R5---~o X~R3 Suitable bases for generating the anion of a compound of formula ~ and 5 include, but are not limited to, sodamide, lithium diisopropylamide (LDA), sodium hydride, potassium ~L~-butoxide, sodium alkoxide, potassium alkoxide, potassium hydride, lithium 2, 2, 6, 6- tetramethylpiperidine, butyl lithium, sec-butyl lithium, ~L~-butyl lithium, and lithium- sodium-, or 10 potassium hexamethyldisilazide. The reaction can be conducted in an aprotic solvent, generally in an ether, such as but not limited to, tetrahydrofuran (THF), dioxane, glyme, diglyme, or diethyl ether. Addition-ally, the reaction can be run in dimethylformamide or dimethylacetamide. However, if the compounds are soluble in a nonpolar solvent, the reaction can be carried out in a hydrocarbon solvent such as hexanes, heptane, cyclohexane, methylcyclohexane, benzene or DM-6617 ~146000 -; 9~ / 0 l ~ 7 6 toluene. Depending on the strength of the base, the reactions can be conducted at a temperature from about -78C to solvent reflux temperature. Instead of running the reaction sequentially, one may at times, add two 5 equivalents of base to the compounds of formula 4, followed by two to three equivalents of the alkylating agent. Alternatively, phase-transfer catalysis conditions may be used, employing a solvent such as benzene, toluene, xylene, dichloromethane, dichloro-10 ethane,or chloroform in conjunction with a quaternaryammonium salt or a quaternary phosphonium salt in the presence of an aqueous base, such as sodium hydroxide or potassium hydroxide. This procedure is described by Bryant and Huhn, in US Patent 4,806,651 and in J.
15 Heterocycl~c Ch~m. 2Q, 771 (1983), which is herein incorporated by reference.
When the alkylation reaction is complete as evidenced by thin layer chromatography, excess anion is decomposed with saturated ammonium chloride solution, and the reaction is taken through an acid- base cycle to remove neutral starting materials. Purification of the basic product generally involves conventional purification techniques such as flash chromatography followed by recrystallization if necessary. The pure 25 base (one spot on thin layer chromatography and analytical HPLC) may be collected as an oil, gum, or amorphous solidi or recrystallized from an appropriate solvent system; or further purified by chromatographic, sublimation, or distillation processes. The compounds may exist as the "free base" or as an acid addition salt formed from pharmaceutically acceptable acids.
Additionally, compounds of formula I may exist as racemates, diastereomeric mixtures, or their optically pure isomers.
Compounds of formula 5 may also be prepared according to Scheme 2. This procedure is particularly useful when R2 is not the same as -CH2-R1 because 5 may be obtained uncontaminated by 4 or the dialkylated 21~6DOO R~rr~ 9~s~
DM-6617 , ~ 4 product. This procedure is fully described by Bryant and Huhn, US Patent 4,806,651, which is herein incorporated by reference.
Scheme 2 Cl Rs_~ CICOCOCI R5--~ ~= AlCb ~R3 R~ 2 R~ 6 R1~

Rs_~ R-CH3 Rs ~ reducbon R~ 7 R~ 8 In addition to the Friedel-Crafts cycloalkylation illustrated by Scheme 1, compounds of the formula 4 may also be prepared by the general "azasulfonium ion"
10 rearrangement methods of Gassman U.S. Patents 3,897,451 (1975), 3,996,264 tl976), and 3,972,894 (1976) which are herein incorporated by reference; see also J. ~m. Chem.
Soc., 96, 5512 (1974), Synthes;s 534 (1981). This route - is shown in Scheme 3.

DM-6617 2 1 4 6 0~0 t~ ~ 4 7 6 Sch~me 3 ~3~ ,H ~ ~CI R~
~R3 X~J~R3 R4 2 R~ 9 SnCI4 R5--~=o ~N Raney X~b al Nkkel ~R3 r4 10 Other representative compounds of this invention can be synthesized by converting one Y group to another.
For example, a compound of formula I which is an ester (Y = Co2R7) may be converted to the corresponding alcohol (Y = CH2OH) of formula I which can be further converted to an ether (Y = oR6) or the "reverse ester"
(Y = CH2ocoR7). For such a case, the ester can be 10 saponified to give the acid (Y = CO2H) which can be reduced to the alcohol. Alternatively, the ester can be directly reduced to the alcohol, which can be subsequently acylated with an acid halide or anhydride, or by coupling the alcohol to an acid using dicyclohexylcarbodiimide, carbonyl diimidazole, or some other coupling agent.
A nitrile can be oxidized to the corresponding amide using the procedure described by Noller, Org. Syn.
Coll. Vol. II, p 586. The same amide can be prepared from the corresponding ester by saponification, activation of carboxyl, and reaction with ammonia or ammonia derivatives. By substituting primary or 2146~a'~ 93/ o^
DM-6617 R ~ ~ v ~ 4 secondary amines for ammonia, other compounds of this invention may be prepared.
The compounds listed in Tables I and II may be used as starting materials for the preparation of compounds of this invention. The listed compounds are either commercially available or disclosed in the literature.
These lists are not comprehensive, and are intended to illustrate the invention, not to limit it. All references in Tables are incorporated by reference.
o Table 1 R5~[~ ~H ~ H

XA~ 3 R5~3dC ~=

X `¢¦ X ~;J~ R3 Cmpd No. St* X a~ Q R3 R4 R5 R5 Reference Pl A O - - - - H H Commercial P2 -A S - - - - H H Commercial P3 A CH2 - - - _ H H Commercial P4 A CH2 - - - _ H F J. Indian Chem.
soc. 38, ~77 (1961~
P5 A CH2 - - - _ F F J. Am. Chem.
soc. 63, 1563 (19 DM-6617 214 6 û ~ ~ ~ 9 3 / o ~ 4 7 ,, P6 A CH2CH2 ~ ~ ~ ~ H H Commercial P7 C S - H2 - - H H Acad. Sci.
Pari3 265, 758 ~1967) P8 C S - O - - H H J . Org . Chem.
36, 2437 (1971 P9 C O - H2 - - H H J . Med. Chem.
33, 2019 (1990); EP
332,364 A2 P10 C CH2CH2 - H2 ~ ~ H H GB 897,052 P11 C CH2 - - - - H H J . Med. Chem 33, 2019 (1990) P12 C CH2 - - - - F H ~
P13 C CH2 - - - - F F " . -P14 D S s H2 H H H
P15 D CH2 s H2 H H OCH3 -P16 D CH2 s H2 H H F - n P17 D CH2 s H2 H H H H GB 1, 394,373 P18 D O s H2 CH3 H CH3 - EP 332,364 A2 P19 C CH2 - H2 ~ ~ H H n P20 C CH2 - H2 ~ ~ H F n P21 C CH2 ~ H2 ~ ~ F F
P22 D CH2 S H2 H H CH3 - GB 1, 394,373 P23 D CH2N-Pr s H2 H CH3 H ~ J. Med. Chem.
34, 3187 (1991) P24 B CH2NH s - H H H - J. Med. Chem.
20, 209 (1977) P25 B CH2NCH3 s - H H H -P26 B CH2NH s - H H C1 P27 B NSO2Ph s - H H H - J. Am. Chem.
Soc. 69, 795 (1947) ~29 D CH2 s H2 =O H H H J . Org . Chem.
55, 560 (1990) P30 D s s O H H H H J . Med. Chem.
22, 1074 (1979 DM-6617 ~ Q~)D

P28 D NH s O H =O H H US 4,087,527 * s indicates single bond St = structure commercial = commercially available T~bl- II

Rs~=

R7 R3 R~ J~

E F
Cmpd.
No. Struct. Q R3 R4 R5 R7 Reference P31 E _ (CH3) 2 H H CH3 Commercial P32 F O (CH3) 2 H H Ph US 4,198, 41-4 --P33 F O (CH3)2 H H CH3 ~
P34 F O (CH3) 2 H CH3 CH3 "
P35 F H,H H H F H EP 332, 364 A2 Struct. = structure commercial = commercially available Analytical data were recorded for the compounds described below using the following general procedures.
Proton NMR spectra were recorded on a Varian FT-NMR
S spectrQmeter (200 MHz or 300 MHz); chemical shifts were recorded in ppm (a) from an internal tetramethylsilane standard in deuterochloroform or deuterodimethylsulfoxide and coupling constants (J) are reported in Hz. Mass spectra (MS) or high resolution mass spectra ~HRMS) were recorded on Finnegan MAT 8230 spectrometer or Hewlett Packard 5988A model spectrometer. Melting points are uncorrected. Boilin~
points are uncorrected.
Reagents were purchased from commercial sources and, where necessary, purified prior to use according the general procedures outlined by D. D. Perrin and w DM-6617 2 1 4 6 ~ Q ~ 9 3 / 0, 4 7 6 L. F. Armarego, ~urification of Laboratory Chemicals, 3rd ed., (New York: Pergamon Press, 1988).
Chromatography was performed on silica gel using the solvent systems indicated below. For mixed solvent s systems, the volume ratios are given. Parts and percentages are by weight unless otherwise specified.
Common abbreviations include: THF (tetrahydrofuran), TBDMS (t-butyl-dimethylsilyl), DMF (dimethylformamide), Hz (hertz) TLC (thin layer chromatography).All 10 temperatures are given in degrees centigrade (C).
The following examples and preparations are for illustrative purposes only and are not to be construed as limiting the invention.

Pre~r~tion 1 ~0 .- ~
Pyrrolo r 3 2,1-kllDhenoxazin-1 2-dione To a solution of phenoxazine (21-.05 g, 0.115 mole) in dichloroethane (125 ml) was added oxalyl chloride (1.05 eq., 0.121 mole, 15.3 g, 10.52 ml) with stirring.
The solution was heated to 50C for one hour, then cooled to room temperature. A solution of aluminum chloride in nitrobenzene (lM, 1 eq., 0.115 mole, 115 ml) was added via addition funnel at room temperature. The solution was stirred for 6 hours, cooled with an ice bath, and quenched with lN HCl and water. More dichloroethane was added, and the organic solution was washed sequentially with water, aq. NaHCO3 and brine, and dried over magnesium sulfate. The dichloroethane was removed via rotary evaporation, and 1000 ml of hexane was added to the resulting dark nitrobenzene DM-6617 21~60a~ 9 3 / O ~ ~ 7 6 solution to give dark violet crystals. The solid was filtered, and dried under vacuum to provide 19.95 g (0.084 mol, 73% yield) of the title compound, mp 218-9 C. MS (NH3/CI) m/e 238 (M+H). Anal. Calcd for s C14H7NO3: C, 70.89; H, 2.97; N, 5.90. Found: C, 70.62;
H, 2.89; N, 5.82.
Prep~r~tion 2 ~ N
~0 ~ r `~
1,?-D;~y~ro-?-(4-~ethenylpyri~;nyl)~yrrolo~3,?,1-10kllphenox~z~n-l(?H)-one To a mixture of the compound from Preparation 1 (2.38 g, 0.010 mole) in acetic anhydride (20 ml) was added acetic acid (2 ml) and 4-picoline (1.75 eq., 0.0175 mole, 1.63 g, 1.7 ml). The mixture was heated to 110 C, and held at this temperature for 45 min., at which time a red precipitate had formed. The reaction - mixture was cooled to room temperature, and poured over ice. The solid was collected and washed with water.
The material was dried under vacuum overnight to obtain 2.94 g (0.0094 mole, 94% yield) of the title compound, mp 240-5C. MS (NH3/CI) m/e 313 (M+H).

Pre~r~tion 3 H /~N
~0 ~

DM-6617 ~14 f~ ~ a ~ ~ fJ 9 3 ~ O - 4 7` 6 1 2-Dlhy~ro-?-(4-pyri~inylmet~yl)~yrrolo r 3 2 1_ kll~heno~zin-1(?~)-one The product from Preparation 2 ~3.0 g, 9.6 mmol) was weighed into a Parr~ shaker bottle, tetrahydrofuran (150 ml) was added, along with 160 mg of 20% Pd(OH)2 on carbon and 200 mg 10% Pd/C. The mixture was shaken under 55 psi of hydrogen for 4 hours, at which time the red color had faded. The reaction mixture was filtered to remove catalyst, and the solvent was removed by 10 rotary evaporation. The residue was purified via column chromatography (silica gel, 5% methanol in ethyl acetate), and the solid was recrystallized from ethyl acetate to give 1.902 g (6.1 mmol, 64% yield) of the title compound, mp 180-2 C. lH-NMR (300 MHz, CDCl3) a 15 3.04 (dd, 1 H, J = 9, 14 Hz), 3.45 (dd, 1 H, J = 5, 14 Hz), 3.88 (dd, 1 H, J = 5, 9 Hz), 6.43 (d, 1 H, J = 7 Hz), 6.70 (d, 1 H, J = 8 Hz), 6.83 (d, 1 H, J = 8 Hz), 6.88 (m, 1 H), 7.00 (m, 2 H), 7.13 (d, 2 H, J = 6 Hz), 8.25 (dd, 1 H, J = 3, 7 Hz), 8.51 (d, 2 H, J = 6 Hz).
MS (NH3/CI) m/e 315 (M+H). Anal. Calcd for C20H14N22:
C, 76.42; H, 4.49; N, 8.91. Found: C, 76.23; H, 4.42, N; 8.81.

FxamDle 193 2s N~ ~N
[~0 ~¢~
2 2-Bis(4-~yri~;nylmethyl)-~yrrolo~3.2.1-kll~henoxazin_ ~ )-one.

To a solution of pyrrolo[3.2.1-kl]phenoxazin-1(2H)-one (486 mg, 2.18 mmol) in toluene (25 ml) was added 4-DM-6617 2 1 ~ 6 ~ 0 ~ , ? ,~ , picolyl chloride hydrochloride (2.2 eq., 4.8 mmol, 787 mg) and benzyltriethylammonium chloride (0.087 eq., 0.19 mmol, 43 mg). While the mixture was stirred at room temperature, 50% aq. sodium hydroxide was added s dropwise. The mixture was heated to 60C for 2 hours, at which time TLC showed that all starting materials had been consumed. Water was added, and the dark brown reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with water, then 10 brine, and the solution was dried over magnesium sulfate. After removal of solvent, the residue was purified via column chromatography (silica gel, 5%
methanol in ethyl acetate), and the solid was recrystallized from dichloromethane/hexane to give 471 15 mg (1.16 mmol, 53% yield) of the title compound, mp 199- ~-200C. MS (NH3/CI) m/e 406 (M+H). 1H-NMR (300 MHz, CDCl3) a 3.12 (d, 2 H, J = 13 Hz), 3.38 (d, 2 H, J = 13 Hz), 6.60 (d, 1 H, J = 8 Hz), 6.76 (m, 1 H), 6.82 (d, 1 H, J = 8 Hz), 6.86 (d, 4 H, J = 6 Hz), 6.90- 6.98 (m, 3 20 H), 8.12 (m, 1 H), 8.34 (d, 4 H, J = 6 Hz). Anal. Cald for C26H1gN3O2 : C, 77.02; H, 4.72; N, 10.36. Found:
C, 76.81; H, 4.62; N, 10.28.

In a similar manner, the following compounds were 25 prepared:
2,3-Dihydro-6,6-bis(4-pyridinylmethyl)-pyrrolo[1,2,3-de]-1,4-benzothiazin-5(6H)-one (Ex. 49):
mp 160-161 C, MS (NH3/CI) m/e 374 (M+H), 1H-NMR (300 MHz, CDCl3) a 2.44 (t, 2 H), 3.11 (d, 2 H, J = 12.9 Hz), 30 3.32 (d, 2 H, J = 12.8 Hz), 3.56 (t, 2 H), 6.79 (d, 4 H, J = 1.5 Hz), 6.91 (m, 3 H), 8.31 (d, 4 H, J = 1.5 Hz).
Anal. Calcd for :C22H19N3OS: C, 70.75; H, 5.13; N, 11.25. Found: C, 70.32; H, 5.01; N, 11.04.
1,1-Bis(4-pyridinylmethyl)-5,6-dihydro-4H-35 pyrollo[3,2,1-ij]quinolin-2(1H)-one (Ex. 1): mp 140-1C, MS (CH4/CI) m/e 356 (M+H), 384 (M+29), 1H-NMR (300 MHz, CDC13) a 1.45 (quin, 2 H, J = 6 Hz), 2.42 (t, 2 H, J = 6 Hz), 3.11 (d, 2 H, J = 13 Hz), 3.26 (t, 2 H, J = 6 2l46n~ R~ 93/ 0? ~ 7 6 DM-6617 - ~

Hz), 3.33 (d, 2 H, J = 13 Hz), 6.82 (d, 4 H, J = 6 Hz), 6.86 (d, 1 H, J = 7 Hz), 6.94 (t, 1 H, J = 7 Hz), 7.07 (d, 1 H, J = 7 Hz), 8.28 (d, 4 H, J = 6 Hz). Anal.
Calcd for C23H21N3O: C, 77.72; H, 5.g6; N, 11.82.
s Found: C, 77.65; H, 5.79; N, 11.77.
2,2-Bis(4-pyridinylmethyl)-pyrrolo[3,2,1-kl]phenothiazin-1(2H)-one (Ex. 169): mp 190-1C, MS
(CH4/CI) m/e 422 (M+H), 450 (M+29), lH-NMR (300 MHz, CDC13) ~ 3.10 (d, 2 H, J = 13 Hz), 3.38 (d, 2 H, J = 13 10 Hz), 6.69 (d, 1 H, J = 7 Hz), 6.77- 7.08 (m, 5 H), 6.86 (d, 4 H, J = 6 Hz), 8.34 (d, 4 H, J = 6 Hz), 8.58 (d, 1 H, J = 8 Hz). Anal. Calcd for C26H1gN3OS: C, 74.09; H, 4.54; N, 9.97. Found: C, 73.87; H, 4.34; N, 9.82.
C,H,N.
2,2-Bis(4-pyridinylmethyl)-6,7-dihydro-indolo[1,7-ab][l]benzapin-1(2H)-one dihydrochloride (Ex. 217): mp 274-6C, MS (CH4/CI) m/e 418 (M+H), 446 (M+29), HRMS:
m/e calcd 417.1841, m/e found 417.1836. Anal. Calcd for C2gH23N3O-2HCl-H2O: C, 66.14; H, 5.35; N, 8.26.
Found: C, 66.14; H, 5.23; N, 8.15.

F.x~n~ple ~ O O

N~
~CN

~

1 2-Dihydro-2-(4-pyri~inylmethyl)-2-(~entanenitrile)_ Dyrrolo~3.2.1-kll~henoxazin-1(2H)-one. hydrohromide To a slurry of sodium hydride (60% in oil, 176 mg, 4.4 mmol) in THF at room temperature was added the compound from Preparation 3 (2.2 mmol, 692 mg), followed by 5-bromovaleronitrile (5 eq., 11 mmol, 1.78 g). One R~ 93~Q^~7,~6 DM-6617 21~ ~ O ~

drop of ethanol was added, and the mixture was stirred at room temperature for 3 days. Methanol was added to decompose excess sodium hydride, and the solvents were removed by rotary evaporation. The residue was s partitioned between ethyl acetate and water, then the product was extracted into lN HCl. The aqueous layer was basified, and the product was extracted into dichloromethane. Subsequent purification by column chromatography provided an oil, 475 mg, 55% yield. MS
10 (NH3/CI) m/e 396 (M+H). lH-NMR (300 MHz, CDCl3) ~ 1.25 (m, 2 H), 1.61 (m, 2 H), 1.88 (m, 1 H), 2.13 (m, 1 H), 2.25 (m, 2 H), 2.99 (d, 1 H, J = 13 Hz), 3.20 (d, 1 H, J
= 13 Hz), 6.68 (d, 1 H, J = 8 Hz), 6.78 (d, 1 H, J = 8 Hz), 6.84 (d, 2 H, J = 6 Hz), 6.86 (m, 1 H), 6.98 (m, 3 15 H), 8.19 (dd, 1 H, J = 2, 7 Hz), 8.31 (d, 2 H, J = 6 Hz). The compound was converted into a salt with hydrobromic acid, mp 239-45 C. Anal. Calcd for C2sH21N3O2-2HBr-0.25H2O: C, 62.44; H, 4.72; N, 8.74.
Found: C, 62.61; H, 4.58; N, 8.64.
By using the methods illustrated in the above examples, compounds in Tables III, IV and V can be prepared.
Table III

R5--~o ~R3 Ex. X Rl R2 R3 R4 a R5 mpC.
No.
1 CH2 4-pyr CH2-(4-pyr) H H s H 140-1 2 CH2 4-pyr CH2-(3-pyr) H H s H

3 CH2 4-pyr CH2-(2-pyr) H H s H

4 CH2 4-pyr CH2-(4-pyrim) H H s H

2 1 ~ 6 0 a ~ ~ 9 3 o DM- 6 6 1 7 R . ~ . 4:

5 CH2 4-pyr CH2-(2-F-4- H H s H
pyr) 6 CH2 4-pyr CH2-(3-CN- H H s H
C6H5) 7 CH2 4-pyr (CH2)3CN H H s H

8 CH2 4-pyr (CH2)4CN H H s H

9 CH2 4-pyr (CH2)sCN H H s H
CH2 4-pyr (CH2)3CO2Et H H s H
11 CH2 4-pyr (CH2)4CO2Et H H s H
12 CH2 4-pyr CH2-Ph H H s H
13 CH2 4-pyrim CH2-(4-pyr) H H s H
14 CH2 4-pyrim CH2-(3-pyr) H H s H
CH2 4-pyrim CH2-(2-pyr) H H s H
16 CH2 4-pyrim CH2- (4-pyrim~ H H s H
17 CH2 4-pyrim CH2-(2-F-4- H H s H
pyr) 18 CH2 4-pyrim CH2-(3-CN- H H s H
C6H5 ) 19 CH2 4-pyrim (CH2)3CN H H s H
CH2 4-pyrim (CH2)4CN H H s H
21 CH2 4-pyrim . (CH2)sCN H H s H
22 CH2 4-pyrim (CH2)3CO2Et H H s H
23 CH2 4-pyrim (CH2)4CO2Et H H s H
24 CH2 4-pyrim CH2-Ph H H s H
25 CH2cH2 4-pyr CH2-(4-pyr) H H s H
26 CH2cH2 4-pyr CH2-(3-pyr) H H s H
27 CH2cH2 4-pyr CH2-(2-pyr) H H s H
28 CH2cH2 4-pyr CH2-(4-pyrim) H H s H
29 CH2cH2 4-pyr CH2-(2-F-4- H H s H
pyr) 30 CH2CH2 4-pyr CH2-(3-CN- H H s H
C6H5 ) 31 CH2CH2 4-pyr (CH2)3CN H H s H
32 CH2cH2 4-pyr (CH2)4CN H H s H
33 CH2CH2 4-pyr (CH2)SCN H H s H
34 CH2CH2 4-pyr (CH2)3CO2Et H H s H
35 CH2CH2 4-pyr (CH2)4CO2Et H H s H

DM-6617 2 1 4 6 ~ 93 / O J ~ 7 6~ i ~

36 CH2cH2 4-pyr CH2-Ph H H s H
37 CH2CH2 4-pyrimCH2-(4-pyr) H H s H
38 CH2cH2 4-pyrimCH2-(3-pyr) H H s H
39 CH2CH2 4-pyrimCH2-(2-pyr) H H s H
40 CH2CH2 4-pyrim CH2-(4-pyrim) H H s H
41 CH2CH2 4-pyrimCH2-(2-F-4- H H s H
pyr) 42 CH2cH2 4-pyrimCH2-(3-CN- H H s H
C 6H5 ) 43 CH2CH2 4-pyrim(CH2)3CN H H s H
44 CH2CH2 4-pyrim(CH2)4CN H H s H
45 CH2CH2 4-pyrim(CH2)sCN H H s H
46 CH2cH2 4-pyrim(CH2)3CO2Et H H s H
47 CH2CH2 4-pyrim(CH2)4CO2Et H H s 48 CH2cH2 4-pyrimCH2-Ph H H s H ;~
49 S 4-pyrCH2-(4-pyr) H H s H 160-1 S 4-pyr CH2-(3-pyr) H H s H
51 S 4-pyr CH2-(2-pyr) H H s H
52 S 4-pyr CH2-(4-pyrim) H H s H
53 S 4-pyr CH2-(2-F-4- H H s H
pyr) 54 S 4-pyr CH2-(3-CN- H H s H
C6H5 ) S 4-pyr (CH2)3CN H H s H
56 S 4-pyr (CH2)4CN H H s H
57 S 4-pyr (CH2)sCN H H s H
58 S 4-pyr (CH2)3CO2Et H H s H
59 S 4-pyr (CH2)4CO2Et H H s H
S 4-pyr CH2-Ph H H s H
61 S 4-pyrim CH2-(4-pyr) H H s H
62 S 4-pyrim CH2-(3-pyr) H H s H
63 S 4-pyrim CH2-(2-pyr) H H s H
64 S 4-pyrim CH2-(4-pyrim) H H s H
S 4-pyrim CH2-(2-F-4- H H s H
pyr) 66 S 4-pyrim CH2-(3-cN- H H s H
C6H5) 21460~3 R`~ 9310^476 DM-6617 0~

67 S 4-pyrim (CH2)3CN H H s H
68 S 4-pyrim (CH2)4CN H H s H
69 S 4-pyrim (CH2)sCN H H s H
S 4-pyrim (CH2)3CO2Et H H s H
71 S 4-pyrim (CH2)4CO2Et H H s H
72 S 4-pyrim CH2-Ph H H s H
73 bond 4-pyr CH2-(4-pyr) H H s H
74 bond 4-pyr CH2-(3-pyr) H H s H
bond 4-pyr CH2-(2-pyr) H H s H
76 bond 4-pyr CH2-(4-pyrim) H H s H
77 bond 4-pyr CH2-(2-F-4- H H s H
pyr) 78 bond 4-pyr CH2-(3-CN- H H s H
C6H5) 79 bond 4-pyr (CH2)3CN H H s H
bond 4-pyr (CH2)4CN H H s H
81 bond 4-pyr (CH2)sCN H H s H
82 bond 4-pyr (CH2)3CO2Et H H s H
83 bond 4-pyr (CH2)4CO2Et H H s H
84 bond 4-pyr CH2-Ph H H s H
bond 4-pyrim CH2-(4-pyr) H H s H
86 bond 4-pyrim CH2-(3-pyr) H H s H
87 bond 4-pyrim CH2-(2-pyr) H H s H
88 bond 4-pyrim CH2-(4-pyrim) H H S H
89 bond 4-pyrim CH2-(2-F-4- H H s H
pyr) bond 4-pyrim CH2-(3-CN- H H s H
C6H5) 91 bond 4-pyrim (CH2)3CN H H s H
92 bond 4-pyrim (CH2)4CN H H s H
93 bond 4-pyrim (CH2)sCN H H s H
94 bond 4-pyrim (CH2)3CO2Et H H s H
bond 4-pyrim (CH2)4CO2Et H H s H
96 bond 4-pyrim CH2-Ph H H s H
97 C=O 4-pyr CH2-(4-pyr) H H s H
98 C=O 4-pyr CH2-(3-pyr) H H s H
99 C=O 4-pyr CH2-(2-pyr) H H s H

_ _ ~

DM-6617 2 ~ ~ 6 ~ 9 3 / ~ 4 7 6 100 C=O 4-pyr CH2-(4-pyrim) H H s H
101 C=O 4-pyr CH2-(2-F-4- H H s H
pyr) 102 C=O 4-pyr CH2-(3-CN- H H s H
C 6H5 ) 103 C=O 4-pyr(CH2)3CN H H s H
104 C=O 4-pyr(CH2)4CN H H s H
lQ5 C=O 4-pyr(CH2)sCN H H s H
106 C=O 4-pyr(CH2)3CO2Et H H s H
107 C=O 4-pyr(CH2)4CO2Et H H s H
108 C=O 4-pyr CH2-~h H H s H
109 C=O 4-pyrim CH2-(4-pyr) H H s H
110 C=O 4-pyrim CH2-(3-pyr) H H s H
111 C=O 4-pyrim CH2-(2-pyr) H H s H
112 C=O 4-pyrim CH2-(4-pyrim) H H s H ;~r~`- ' 113 C=O 4-pyrim CH2-(2-F-4- H H s H
pyr) 114 C=O 4-pyrim CH2-(3-CN- H H s H
C 6H5 ) 115 C=O 4-pyrim (CH2)3CN H H s H
116 C=O 4-pyrim (CH2)4CN H H s H
117 C=O 4-pyrim (CH2)sCN H H s H
118 C=O 4-pyrim(CH2)3CO2Et H H s H
119 C=O 4-pyrim(CH2)4CO2Et H H S H
120 C=O 4-pyrim CH2-Ph H H s H
121 CH2NPr 4-pyr CH2-(4-pyr) H CH3 s H
122 CH2NPr 4-pyr CH2-(3-pyr) H CH3 s H
123 CH2NPr 4-py~ CH2-(2-pyr) H CH3 s H
124 CH2NPr 4-pyr CH2-(4-pyrim) H CH3 s H
125 CH2NPr 4-pyr CH2-(2-F-4- H CH3 S H
pyr) 126 CH2NPr 4-pyr CH2-(3-CN- H CH3 S H
C6H5) 127 CH2NPr 4-pyr (CH2) 3CN H CH3 s H
128 CH2NPr 4-pyr (CH2) 4CN H CH3 s H
129 CH2NPr 4-pyr (CH2) 5CN H CH3 s H
130 CH2NPr 4-pyr (CH2)3CO2Et H CH3 s H

DM--6617 2 1 ~ 6 R G ; r ~ q ~ + ~ 6 131 CH2NPr 4-pyr(CH2)4CO2Et H CH3 s H
132 CH2NPr 4-pyrCH2-Ph H CH3 s H
133 CH2NPr 4-pyrim CH2-(4-pyr) H CH3 s H
134 CH2NPr 4-pyrim CH2-(3-pyr) H CH3 s H
135 CH2NPr 4-pyrim CH2-(2-pyr) H CH3 s H
136 CH2NPr 4-pyrim CH2-(4-pyrim) H CH3 s H
137 CH2NPr 4-pyrim CH2-(2-F-4- H CH3 s H
pyr) ~-138 CH2NPr 4-pyrim CH2-(3-CN- H CH3 s H
C 6H5 ) 139 CH2NPr 4-pyrim (CH2)3CN H CH3 s H
140 CH2NPr 4-pyrim (CH2)4CN H CH3 s H
141 CH2NPr 4-pyrim (CH2)sCN H CH3 s H
142 CH2NPr 4-pyrim (CH2)3CO2Et H CH3 s H
143 CH2NPr 4-pyrim (CH2)4CO2Et H CH3 s H - ~-144 CH2NPr 4-pyrim CH2-Ph H CH3 s 145 S 4-pyr CH2-(4-pyr) Ph H s H
146 S 4-pyr CH2-(3-pyr) Ph H s H
147 S 4-pyr CH2-(2-pyr) Ph H s H
148 S 4-pyr CH2-(4-pyrim) Ph H s H
149 S 4-pyrCH2-(2-F-4- Ph H s H
pyr) 150 S 4-pyrCH2-(3-CN- Ph H s H
C 6H5 ) 151 S 4-pyr (CH2)3CN Ph H s H
152 S 4-pyr (CH2)4CN Ph H s H
153 S 4-pyr(CH2)sCN Ph H s H
154 S 4-pyr.(CH2)3CO2Et Ph H s H
155 S 4-pyr(CH2)4CO2Et Ph H s H
156 S 4-pyr CH2-Ph Ph H s H
157 S 4-pyrim CH2-(4-pyr) Ph H d H
158 S 4-pyrim CH2-(3-pyr) Ph H d H
159 S 4-pyrim CH2-(2-pyr) Ph H d H
160 S 4-pyrim CH2-(4-pyrim) Ph H d H
161 S 4-pyrim CH2-(2-F-4- Ph H d H
pyr) ~1~6000 ~ 93/ 0~47 6 DM-6617 ~ ~

162 S 4-pyrim CH2-(3-CN- Ph H d H
C6H5 ) 163 S 4-pyrim(CH2) 3CN Ph H d H
164 S 4-pyrim(CH2) 4CN Ph H d H
165 S 4-pyrim(CH2) 5CN Ph H d H
166 S 4-pyrim(CH2)3C02Et Ph H d H
167 S 4-pyrim(CH2)4CO2Et Ph H d H
168 S 4-pyrim CH2-Ph Ph H d H

DM-6617 21460~0 9~/ G ~ 7 6 T~bl~ rv RS_~o X ~1 1 Ex. X Rl R2 R5 R5' mpC.
No.
169 S 4-pyrCH2-(4-pyr) H H 190-1 170 S 4-pyr CH2-(3-pyr) H H
171 S 4-pyr CH2-(2-pyr) H H
172 S 4-pyr CH2-(4-pyrim) H H ~~
173 S 4-pyr CH2-(2-F-4- H H
pyr) 174 S 4-pyr CH2-(3-CN- H H
C6H5 ) 175 S 4-pyr(CH2)3CN H H
176 S 4-pyr(CH2)4CN H H
177 S 4-pyr(CH2)sCN H H
178 S 4-pyr(CH2)3CO2Et H H
179 S 4-pyr(CH2)4CO2Et H H
180 S 4-pyr CH2-Ph H H
181 S 4-pyrim CH2-(4-pyr) H H
182 S 4-pyrim CH2-(3-pyr) H H
183 S 4-pyrim CH2-(2-pyr) H H
184 S 4-pyrim CH2-(4-pyrim) H H
185 S 4-pyrim CH2-(2-F-4- H H
pyr) 186 S 4-pyrim CH2-(3-CN- H H
C6H5 ) 187 S 4-pyrim (CH2)3CN H H
188 S 4-pyrim (CH2)4CN H H
189 S 4-pyrim(CH2)sCN H H

DM-6617 21 4 6 ~ O O
~ J ~

190 S 4-pyrim(CH2)3CO2Et H H
191 S 4-pyrim(CH2)4CO2Et H H
192 S 4-pyrim CH2-Ph H H
193 O 4-pyr CH2-(4-pyr) H H199-200 194 O 4-pyr CH2-(3-pyr) H H
195 O 4-pyr CH2-(2-pyr) H H
196 O 4-pyr CH2-(4-pyrim) H H
197 O4-pyr CH2-(2-F-4- H H
pyr) 198 O 4-pyr CH2-(3-CN- H H
C6H5 ) 199 O 4-pyr (CH2)3CN H H
200 O 4-pyr (CH2)4CN H H239-45 (-HBr) 201 O 4-pyr (CH2)sCN H H ~~~
202 O 4-pyr (CH2)3CO2Et H H
203 4-pyr (CH2)4CO2Et H H
204 O 4-pyr CH2-Ph H H
205 4-pyrim CH2-(4-pyr) H H
206 O 4-pyrim CH2-(3-pyr) H H
207 4-pyrim CH2-(2-pyr) H H
208 O 4-pyrim CH2-(4-pyrim) H H
209 O4-pyrim CH2-(2-F-4- H H
pyr) 210 O4-pyrim CH2-(3-CN- H H
C6H5 ) 211 O 4-pyrim (CH2)3CN H H
212 O 4-pyrim (CH2)4CN H H
213 o 4-pyrim (CH2)sCN H H
214 O 4-pyrim(CH2)3CO2Et H H
215 O 4-pyrim(CH2)4CO2Et H H
216 O 4-pyrim CH2-~h H H
217 CH2CH2 4-pyr CH2-(4-pyr) H H 274-6 (-2HCl) 218 CH2cH2 4-pyr CH2-(3-pyr) H H
219 CH2cH2 4-pyr CH2-(2-pyr) H H
220 CH2CH2 4-pyr CH2-(4-pyrim) H H

DM-6617 2146000 c 93/ 0, ~ 7 6 221 CH2cH2 4-pyr CH2-(2-F-4- H H
pyr) 222 CH2cH2 4-pyr CH2-(3-CN- H H
C 6H5 ) 223 CH2cH2 4-pyr (CH2)3CN H H
224 CH2cH2 4-pyr (CH2)4CN H H
225 CH2cH2 4-pyr (CH2)sCN H H
226 CH2cH2 4-pyr (CH2)3CO2Et H H
227 CH2cH2 4-pyr (CH2)4CO2Et H H
228 CH2cH2 4-pyr CH2-Ph H H
229 CH2cH2 4-pyrimCH2-(4-pyr) H H
230 CH2cH2 4-pyrimCH2-(3-pyr) H H
231 CH2cH2 4-pyrimCH2-(2-pyr) H H
232 CH2CH2 4-pyrim CH2-(4-pyrim) H H
233 CH2cH2 4-pyrimCH2-(2-F-4- H H
pyr) 234 CH2cH2 4-pyrimCH2-(3-CN- H H
C6H5) 235 CH2cH2 4-pyrim(CH2)3CN H H
236 CH2cH2 4-pyrim(CH2)4CN H H
237 CH2cH2 4-pyrim(CH2)sCN H H
238 CH2cH2 4-pyrim(CH2)3CO2Et H H
239 CH2cH2 4-pyrim(CH2)4CO2Et H H
240 CH2cH2 4-pyrimCH2-Ph H H
241 CH2 4-pyrCH2-(4-pyr) H H
242 CH2 4-pyrCH2-(3-pyr) H H
243 CH2 4-pyrCH2-(2-pyr) H H
244 CH2 4-pyrCH2-(4-pyrim) H H
245 CH2 4-pyr CH2-(2-F-4- H H
pyr) 246 CH2 4-pyr CH2-(3-CN- H H
C6H5) 247 CH2 4-pyr(CH2)3CN H H
248 CH2 4-pyr(CH2)4CN H H
249 CH2 4-pyr(CH2)sCN H H
250 CH2 4-pyr(CH2)3CO2Et H H
251 CH2 4-pyr(CH2)4CO2Et H H

DM-6617 2146000 R~ ~ ., ^ r ' ~?4 - 252 CH2 4-pyrCH2-Ph HH
253 CH2 4-pyrim CH2-(4-pyr) H H
254 CH2 4-pyrim CH2-(3-pyr) H H
255 CH2 4-pyrim CH2-(2-pyr) H H
256 CH2 4-pyrim CH2-(4-pyrim) H H
257 CH2 4-pyrim CH2-(2-F-4- H H
pyr) 258 CH2 4-pyrim CH2-(3-CN- H H
C6Hs) 259 CH2 4-pyrim (CH2)3CN H H
260 CH2 4-pyrim (CH2)4CN H H
261 CH2 4-pyrim (CH2)sCN H H
262 CH2 4-pyrim (CH2)3CO2Et H H
263 CH2 4-pyrim (CH2)4C02Et H H
264 CH2 4-pyrim CH2-Ph H H r ~ ~
265 NCH3 4-pyr CH2-(4-pyr) H H
266 NCH3 4-pyrCH2-(3-pyr) H H
267 NCH3 4-pyrCH2-(2-pyr) H H
268 NCH3 4-pyrCH2-(4-pyrim) HH
269 NCH3 4-pyrCH2-(2-F-4- HH
pyr) 270 NCH3 4-pyrCH2-(3-CN- HH
C6H5) 271 NCH3 4-pyr(CH2)3CN HH
272 NCH3 4-pyr(CH2)4CN HH
273 NCH3 4-pyr (CH2)sCN HH
274 NCH3 4-pyr (CH2)3CO2Et HH
275 NCH3 4-pyr (CH2)4CO2Et HH
276 NCH3 4-pyr CH2-Ph HH
277 NCH3 4-pyrimCH2-(4-pyr) HH
278 NCH3 4-pyrimCH2-(3-pyr) HH

279 NCH3 4-pyrimCH2-(2-pyr) HH
280 NCH3 4-pyrim CH2-(4-pyrim) HH
281 4-pyrlm CH2-(2-F-4- HH
pyr) 282 NCH3 4-pyrim CH2-(3-CN- HH
C6H5) DM-6617 2 1 4 6 0 ~ O

283 NCH3 4-pyrim(CH2)3CN H H
284 NCH3 4-pyrim(CH2)4CN H H
285 NCH3 4-pyrim(CH2)sCN H H
286 NCH3 4-pyrim(CH2)3CO2Et H H
287 NCH3 4-pyrim(CH2)4CO2Et H H
288 NCH3 4-pyrim CH2-Ph H H
289 bond 4-pyr CH2-(4-pyr) H H
290 bond 4-pyr CH2-(3-pyr) H H
291 bond 4-pyr CH2-(2-pyr) H H
292 bond 4-pyr CH2-(4-pyrim) H H
293 bond 4-pyr CH2-(2-F-4- H H
pyr) 294 bond 4-pyr CH2-(3-CN- H H
C 6H5 ) 295 bond 4-pyr (CH2)3CN H H
296 bond 4-pyr (CH2)4CN H H
297 bond 4-pyr (CH2)sCN H H
298 bond 4-pyr(CH2)3CO2Et H H
299 bond 4-pyr(CH2)4CO2Et H H
300 bond 4-pyr CH2-Ph H H
301 bond 4-pyrim CH2-(4-pyr) H H
302 bond 4-pyrim CH2-(3-pyr) H H
303 bond 4-pyrim CH2-(2-pyr) H H
304 bond 4-pyrim CH2-(4-pyrim) H H
305 bond 4-pyrim CH2-(2-F-4- H H
pyr) 306 bond 4-pyrim CH2-(3-CN- H H
C 6H5 ) 307 bond 4-pyrim (CH2)3CN H H
308 bond 4-pyrim (CH2)4CN H H
309 bond 4-pyrim (CH2)sCN H H
310 bond 4-pyrim (CH2)3CO2Et H H
311 bond 4-pyrim (CH2)4CO2Et H H
312 bond 4-pyrim CH2-Ph H H
313 C=O 4-pyr CH2-(4-pyr) H H
314 C=O 4-pyr CH2-(3-pyr) H H
315 C=O 4-pyr CH2-(2-pyr) H H

DM-6617 Z146000 ~C ~ ~ .794 316 C=O 4-pyr CH2-(4-pyrim) H H
317 C=O 4-pyr CH2-(2-F-4- H H
pyr) 318 C=O 4-pyr CH2-(3-CN- H H
C6H5) 319 C=O 4-pyr(CH2)3CN H H
320 C=O 4-pyr(CH2)4CN H H
321 C=O 4-pyr(CH2)sCN H H
322 C=O 4-pyr(CH2)3CO2Et H H
323 C=O 4-pyr(CH2)4CO2Et H H
324 C=O 4-pyr CH2-Ph H H
325 C=O 4-pyrim CH2-(4-pyr) H H
326 C=O 4-pyrim CH2-(3-pyr) H H
327 C=O 4-pyrim CH2-(2-pyr) H H
328 C=O 4-pyrim CH2-(4-pyrim) H H
329 C=O 4-pyrim CH2-(2-F-4- H H
pyr) 330 C=O 4-pyrim CH2-(3-CN- H H
C 6H5 ) 331 C=O 4-pyrim (CH2)3CN H H
332 C=O 4-pyrim (CH2)4CN H H
333 C=O 4-pyrim (CH2)sCN H H
334 C=O 4-pyrim (CH2)3CO2Et H H
335 C=O 4-pyrim (CH2)4CO2Et H H
336 C=O 4-pyrim CH2-Ph H H

~14 60 0 ~ `~ 9 3 / 0 . i 7 6 T~bl- V
R1~

R5--~o R7~R3 Ex. Rl R2 R3 R4 R5 R7 mpC
No.
337 4-pyrCH2-(4-pyr) (CH3)2 H H Ph 338 4-pyrCH2-(3-pyr) (CH3)2 H H Ph 339 4-pyrCH2-(2-pyr) (CH3)2 H H Ph 340 4-pyrCH2-(4-pyrim) (CH3)2 H H Ph 341 4-pyr CH2-(2-F-4-pyr) (CH3)2 H H Ph 342 4-pyr CH2-(3-CN-C6Hs) (CH3)2 H H Ph 343 4-pyr(CH2)3CN (CH3)2 H H Ph 344 4-pyr(CH2)4CN (CH3)2 H H Ph 345 4-pyr(CH2)sCN (CH3)2 H H Ph 346 4-pyr(CH2)3CO2Et (CH3)2 H H Ph 347 4-pyr(CH2)4CO2Et (CH3)2 H H Ph 348 4-pyrCH2-Ph (CH3)2 H H Ph 349 4-pyrimCH2-(4-pyr) (CH3)2 H H Ph 350 4-pyrimCH2-(3-pyr) (CH3)2 H H Ph 351 4-pyrimCH2-(2-pyr) (CH3)2 H H Ph 352 4-pyrimCH2-(4-pyrim) (CH3)2 H H Ph 353 4-pyrim CH2-~(2-F-4-pyr) (CH3)2 H H Ph 354 4-pyrim CH2-(3-cN-c6Hs) (CH3)2 H H Ph 355 4-pyrim(CH2)3CN (CH3)2 H H Ph 356 4-pyrim(CH2)4CN (CH3)2 H H Ph 357 4-pyrim(CH2)sCN (CH3)2 H H Ph 358 4-pyrim(CH2)3CO2Et (CH3)2 H H Ph 359 4-pyrim(CH2)4CO2Et (CH3)2 H H Ph 360 4-pyrimCH2-Ph (CH3)2 H H Ph 361 4-pyrCH2-(4-pyr) (CH3)2 H H CH
362 4-pyrCH2-(3-pyr) (CH3)2 H H CH

2 ~ , 9 3 / 0 `., 4 7 6 - DM-6617 ~ ~ ~ 94 3634-pyr CH2-(2-pyr) (CH3) 2 H H CH3 3644-pyr CH2-(4-pyrim) (CH3) 2 H H CH3 3654-pyr CH2-(2-F-4-pyr) (CH3) 2 H H CH3 3664-pyr CH2-(3-cN-c6Hs) tCH3) 2 H H CH3 3674-pyr (CH2)3CN (CH3) 2 H H CH3 3684-pyr (CH2)4CN (CH3~ 2 H H CH3 3694-pyr (CH2)sCN (CH3) 2 H H CH3 3704-pyr (CH2)3CO2Et (CH3) 2 H H CH3 3714-pyr (CH2)4CO2Et (CH3) 2 H H CH3 3724-pyr CH2-Ph (CH3) 2 H H CH3 3734-pyrim CH2-(4-pyr) (CH3) 2 H H CH3 3744-pyrim CH2-(3-pyr) (CH3) 2 H H CH3 375 4-pyrimCH2-(2-pyr) (CH3)2 H H CH3 3764-pyrim CH2-(4-pyrim) (CH3) 2 H H CH3 3774-pyrim CH2-(2-F-4-pyr) (CH3) 2 H H CH3 3784-pyrim CH2-(3-cN-c6Hs) (CH3) 2 H H CH3 3794-pyrim (CH2)3CN (CH3) 2 H H CH3 3804-pyrim (CH2)4CN (CH3) 2 H H CH3 3814-pyrim (CH2)sCN (CH3) 2 H H CH3 3824-pyrim (CH2)3CO2Et (CH3) 2 H H CH3 3834-pyrim (CH2)4CO2Et (CH3) 2 H H CH3 3844-pyrim CH2-Ph (CH3) 2 H H CH3 3854-pyr CH2-(4-pyr) (CH3)2 H CH3 CH3 3864-pyr CH2-(3-pyr) (CH3) 2 H CH3 CH3 3874-pyr CH2-(2-pyr) (CH3) 2 H CH3 CH3 3884-pyr CH2-(4-pyrim) (CH3) 2 H CH3 CH3 3894-pyr CH2-(2-F-4-pyr) (CH3)2 H CH3 CH3 3904-pyr CH2-(3-cN-c6Hs) (CH3)2 H CH3 CH3 3914-pyr (CH2)3CN (CH3)2 H CH3 CH3 3924-pyr (CH2)4CN (CH3)2 H CH3 CH3 3934-pyr (CH2)sCN (CH3)2 H CH3 CH3 3944-pyr (CH2)3CO2Et (CH3)2 H CH3 CH3 3954-pyr (CH2)4CO2Et (CH3)2 H CH3 CH3 3964-pyr CH2-Ph (CH3)2 H CH3 CH3 397 4-pyrim CH2-(4-pyr) (CH3)2 H CH3 CH3 398 4-pyrim CH2-(3-pyr) (CH3)2 H CH3 CH3 399 4-pyrim CH2-(2-pyr) (CH3)2 H CH3 CH3 DM--6617 2 1 4 ~ 9 0 0 P~J r 9 3 / O 4 7 6 400 4-pyrim CH2-(4-pyrim) (CH3)2 H CH3 CH3 401 4-pyrim CH2-(2-F-4-pyr) (CH3)2 H CH3 CH3 402 4-pyrim CH2-(3-CN-C6Hs) (CH3)2 H CH3 CH3 403 4-pyrim(CH2)3CN (CH3)2 H CH3 CH3 404 4-pyrim(CH2)4CN (CH3)2 H CH3 CH3 405 4-pyrim(CH2)sCN (CH3)2 H CH3 CH3 406 4-pyrim(CH2)3CO2Et (CH3)2 H CH3 CH3 407 4-pyrim(CH2)4CO2Et (CH3)2 H CH3 CH3 408 4-pyrimCH2-Ph (CH3)2 H CH3 CH3 409 4-pyr CH2-(4-pyr) H H F H
410 4-pyr CH2-(3-pyr) H H F H
411 4-pyr CH2-(2-pyr) H H F H
412 4-pyr CH2-(4-pyrim) H H F H
413 4-pyr CH2-(2-F-4-pyr) H H F H
414 4-pyr CH2-(3-cN-c6Hs) H H F H
415 4-pyr(CH2)3CN H H F H
416 4-pyr(CH2)4CN H H F H
417 4-pyr(CH2)sCN H H F H
418 4-pyr(CH2)3CO2Et H H F H
419 4-pyr(CH2)4CO2Et H H F H
420 4-pyr CH2-Ph H H F H
421 4-pyrim CH2-(4-pyr) H H F H
422 4-pyrim CH2-(3-pyr) H H F H
423 4-pyrim CH2-(2-pyr) H H F H ~
424 4-pyrim CH2-(4-pyrim) H H F H
425 4-pyrim CH2-(2-F-4-pyr) H H F H
426 4-pyrim CH2-(3-cN-c6Hs) H H F H
427 4-pyrim.(CH2)3CN H H F H
428 4-pyrim(CH2)4CN H H F H
429 4-pyrim(CH2)sCN H H F H
430 4-pyrim(CH2)3CO2Et H H F H
431 4-pyrim(CH2)4CO2Et H H F H
432 4-pyrimCH2-Ph H H F H

21~00~ ~r 93/ 0~J 4 7 6 DM-6617 ~ 4 Bioch~cal Te t Proc~ r~

Neurotransmitt~r rele~se assay: The neurotransmitter release activities of the compounds in this invention s were determined as reported in Drll~ Develop~ent Rese~rch, 1~, 285-300 (1990) and is a modification of the procedure described by Mulder, et al., Rr~ln Res., 70, 372 (1974). These publications are herein incorporated by reference.
0 Male Wistar rats (Charles River) weighing 17S-200 grams were used. The rats were housed for at least seven days before the experiment in animal facility under 12/12 hour light/dark cycle. Deionized water and standard rat chow (Purina) were available ad l ibitum .
Rats were decapitated and brains were removed immediately. Slices (0.3 mm thick) from the parietal cortex were prepared manually using a recessed Lucite guide. Slices were subsequently cut into 0.25 x 0.25 mm squares with a McIlwain tissue chopper.
Cerebral cortical slices (approximately 100 mg wet weight) were incubated in 10 ml Krebs-Ringer medium (KR) containing NaCl (116 mM), KCl (3 mM), CaCl2 (1.3 mM), MgCl2 (1.2 mM), KH2PO4 (1.2 mM), Na2SO4 (1.2 mM), NaHCO3 (25 mM) and glucose (11.0 mM) to whi-ch 10 mCi 3H-choline (specific activity approximately 80 uCi/mM; Du Pont-NEN) and 10 nmol unlabeled choline had been added to give a final concentration of 1 mM. The brain preparations were incubated for 30 minutes at 37C under a steady flow of 95% 02/5% CO2. Under these conditions, part of the 30 radioactive choline taken up by the preparation was converted into radioactive acetylcholine (ACh) by the cholinergic nerve endings stored in synaptic vesicles, and released upon depolarization by high potassium ion (K+) containing media.
3s After labeling of the ACh stores, the slices were washed three times with non-radioactive KR medium and transferred to a superfusion apparatus to measure the drug effects on ACh release. The superfusion apparatus DM-6617 ~ 1 ~ 6 ~ 9 3 / C ~ 7 6 consisted of 10 thermostated glass columns of 5 diameters that were provided with GF/F glass fiber filters to support the slices (approximately 10 mg tissue/column). Superfusion was carried out in KR-medium 5 (O . 3 ml/min.) containing 10 mM hemicholinium-3 (HC-3).
The HC-3 prevents the reuptake of choline formed during the superfusion from phospholipids and released ACh, which would be converted into unlabeled ACh and released -in preference to the pre-formed labeled ACh. The medium 10 was delivered by a 25-channel peristaltic pump (Ismartec by Brinkman) and warmed to 37C in a thermostated stainless steel coil before entering the superfusion column. Each column was provided with a 4-way slider value (Beckmann Instruments) which allowed rapid change 15 of low to high K+/KR-medium, and with two 10-channel 3- ~r'~ ' way values that were used to change from drug-free to drug-containing low and high K+/KR-medium.
After 15 min. of washout of non-specifically bound radioactivity, collection of 4 min. fractions was initiated. After three 4 min. collections, the original medium was changed to a KR-medium in which the KCl concentration has been increased to 25 mM (high K+ -KR
medium; S1). Depolarization-induced stimulation of release by high K+/KR-medium lasted 4 min. Drug free low 25 and high K+/KR-media were then substituted by drug- and vehicle-containing low- and high-K+/KR medium, and superfusion was continued for three 4 min. collections with low K+/KR-medium, one 4 min. collection with high K+/KR-medium (S2), and two 4 min. collections with low-30 K+/KR-medium.
Drug was added to the media by 100-fold dilutions of appropriate concentrations of the drug (in 0.9%
saline) with either low- or high-K+/KR-medium.
All superfusion fractions were collected in liquid 35 scintillation counting vials. After superfusion, the slices were removed from the superfusion columns and extracted with 1.0 ml of O.lN HCl. Liquiscint (NEN) scintillation cocktail (12 ml) was added to superfusion 21~B~00 ~- :` 93~ 0~ 'S 7 6 - DM-6617 ;; ! .` . `

fractions and extracts, and the samples were counted in a Packard Tricarb Liquid Scintillation Counter. No corrections were made for quenching.
The ratio of S2/S1 (as compared to controls where s no drug was present during S2) was a measure of the ability of the drug to enhance or depress stimulus-induced acetylcholine release. Per cent acetylcholine (ACh) enhanced release caused by 10 mM of drug using this assay are shown in Table 5.

Table 5 % Ach release Ex. No. at 10 mM

~tility The foregoing test results suggest that the S compounds of this invention have utility in the treatment of cognitive disorders and/or neurological function deficits and or mood and mental disturbances in patients suffering from nervous system disorders like Alzheimer's Disease, Parkinson's Disease, senile 20 dementia, multi-infarct dementia, Huntington's disease, mental retardation, Myasthenia Gravis, etc. The above-described in vitro assay is recognized as aiding in the identification of drugs useful in the treatment of cognitive disorders and/or neurological function 25 deficits and or mood and mental disturbances in patients suffering from nervous system disorders like Alzheimer's Disease, Parkinson's Disease, senile dementia, multï-infarct dementia, Huntington's disease, mental retardation, Myasthenia Gravis, etc. Cook et al., ~L~

21~60au - $3/ ~ , 7 ~
DM-6617 ~ C ¦ C ~ 4 DeveloDment Rese~rch, 1~, 301-304 (19gO), Nickolson et al., Drug Develop~ent Research, 1~, 285-300 (1990) and DeNoble et al., ~h~rm~cology Rioch~m;stry & Beh~yior, ~, 957-961 (1990), all have shown via the above-described in vitro assay that the drug DuP 996, whichhas the chemical name 3,3-bis(4-pyridinylmethyl)-1-phenylindolin-2-one (linopirdine), is useful in the treatment of cognition dysfunction.
n ~
o Compounds of this invention can be administered to treat said deficiencies by means that produces contact of the active agent with the agent's site of action in the body of a mammal. The compounds can be administered by any conventional means available for use in 15 conjunction with pharmaceuticals either as individual therapeutic agent or in combination of therapeutic agents. They can be administered alone, but are generally administered with a pharmaceutical carrier selected on the basis of the chosen route of 20 administration and standard pharmaceutical practice.
The dosage administered will vary depending on the use and known factors such as pharmacodynamic character of the particular agent, and its mode and route of administrationi the recipient's age, weight, and health;
25 nature and extent of symptoms; kind of concurrent treatment; frequency of treatment; and desired effect.
For use in the treatment of said diseases or conditions, the compounds of-this lnvention can be orally administered daily at a dosage of the active ingredient of 0.002 to 200 mg/kg of body weight. Ordinarily, a dose of 0.01 to 10 mg/kg in divided doses one to four times a day, or in sustained release formulation was effective in obtaining the desired pharmacological effect.
Dosage forms (compositions) suitable for 35 administration contain from about 1 mg to about 100 mg of active ingredient per unit. In these pharmaceutical compositions, the active ingredient will ordinarily be 21~0~ ~,J ~ `. 93/ 0`- ' ~ 6 present in an amount of about 0.5 to 95% by weight based on the total weight of the composition.
The active ingredient can be administered orally is solid dosage forms, such as capsules, tablets and s powders; or in liquid forms such as elixirs, syrups, and/or suspensions. The compounds of this invention can also be administered parenterally in sterile liquid dose formulations.
Gelatin capsules can be used to contain the active 10 ingredient and a suitable carrier such as but not limited to lactose, starch, magnesium stearate, steric acid, or cellulose derivatives. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication l-over a period of time. Compressed tablets can be sugar-coated or film-coated to mask any unpleasant taste, or used to protect the active ingredients from the atmosphere, or to allow selective disintegration of the tablet in the gastrointestinal tract.
Liquid dose forms for oral administration can contain coloring of flavoring agents to increase patient acceptance.
In general, water, pharmaceutically acceptable 2s oils, saline, aqueous dextrose (glucose), and related sugar solutions and glycols, such as propylene glycol or polyethylene glycol, are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, butter substances. Antioxidizing agents, such as sodium bisulfite! sodium sulfite, or ascorbic acid, either alone or in combination, are suitable stabilizing agents. Also used are citric acid 3s and its salts, and EDTA. In addition, parenteral solutions can contain preservatives such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.

~146~ 93/ G^~ 7 6 DM-6617 ~ ' ' ~
--~ V ~ `J

Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciencesn, A. Osol, a standard reference in the field.
Useful pharmaceutical dosage-forms for s administration of the compounds of this invention can be illustrated as follows:

C~Ds-~les A large number of units capsules are prepared by 10 filling standard two-piece hard gelatin capsules each with 100 mg of powdered active ingredient, 150 mg lactose, 50 mg cellulose, and 6 mg magnesium stearate.

Soft Gelatin C~Dsules A mixture of active ingredient in a digestible oil such as soybean, cottonseed oil, or olive oil is prepared and injected by means of a positive displacement was pumped into gelatin to form soft gelatin capsules containing 100 mg of the active ingredient. The capsules were washed and dried.

T~hlets A large number of tablets are prepared by conventional procedures so that the dosage unit was 100 mg active ingredient, 0.2 mg of colloidal silicon dioxide, 5 mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11 mg of starch, and 98.8 mg lactose. Appropriate coatings may be applied to increase palatability or delayed adsorption.
The compounds of this invention may also be used as reagents or standards in the biochemical study of neurological function, dysfunction, and disease.

Claims (12)

What is claimed is:

1. A compound of formula:

or pharmaceutically acceptable salts thereof, wherein:

X is a single bond, O, S, SO, SO2, CH2, CH2CH2, CH=CH, C=O, C(R7)(OR6), CH(OR6), -CONR6-, -NR6CO-, -CH2-NR6-, -NR6-CH2-, NR6, -C(R7)=N-, -CH(R7)-N(R6)-, or -CR7-, -CH- when a is a single bond and b is a double bond;

a and b are each single or double bonds, provided that a is a single bond when b is a double bond, and that b is a single bond when a is double bond, and that b is a double bond when X is -CH- or -CR7- and when a is a single bond;

R1 is 2-, 3- or 4-pyridyl, or 4-pyrimidinyl;

R2 is -(CH2)m-W, wherein m = 1-4, and W is selected from the group:
(a) 2-, 3- or 4-pyridyl, (b) 2-, 4-, or 5-pyrimidinyl, (c) 2-pyrazinyl, (d) 3- or 4-pyridazinyl, (e) 3- or 4-pyrazolyl, (f) 2- or 3-furyl, (g) 2- or 3-tetrahydrofuranyl, (h) 2- or 3-thienyl, (i) 3-indolyl, (j) aryl unsubstituted or substituted with 1-3 R5, (k) 2-fluoro-4-pyridyl, or R2 is -(CH2)n-Y, wherein n = 1-6, and Y is selected from the group:
-CH=CHCO2R7, -CH=CHCOR7, -CH=CHR7, -CH=C(R7)2, -CH=CH2, -C?CCO2R7, -C?CCOR7, -C?CR7, or -C?CH, F, Cl, Br, OR6, N(R6)2, CO2H, CO2R7, CONHR7, NHCHO, CONHR6, CON(R7)2, CN, -OCOR7, COR7, CHO, SR7, SOR7, SO2R7 or NO2;

R3 and R4 are each independently selected from the group:
H, alkyl of 1-6 carbons, alkenyl of 2-6 carbons, alkynyl of 2-6 carbons; cycloalkyl of 3-7 carbons, cycloalkylalkyl of 3-10 carbons, aryl unsubstituted or substituted with 1-3 R5, alkaryl of 1-10 carbons, provided that when a is a single bond R3 is =O, =CH2, =CH(R7), =C(R7)2, or (CH3)2, and provided that when a and b are each a single bond, then R3 and R4 are each independently selected from the group: =O, =CH2, =CH(R7), =C(R7)2, (CH3)2 ; or R3 and R4, taken together may form a saturated or unsaturated carbocyclic or heterocyclic ring, unsubstituted or substituted with 1-2 R5 substituents;

R5 is selected from the group:
H, alkyl of 1-6 carbons, aryl unsubstituted or substituted with 1-3 R8, alkaryl of 1-10 carbons, F, Cl, Br, I, OR6, NHR6, N(R6)2, CO2H, CO2R7, CoNHR7, CON(R7)2, CN, COR7, CHO, SR7, SOR7, SO2R7, NO2 or -CH=CH-CH=CH- which attaches to an adjacent carbon atom forming a fused ring;

R6 is independently selected at each occurrence from the group:
H, alkyl of 1-6 carbons, aryl unsubstituted or substituted with 1-3 R8, alkaryl of 1-10 carbons, -SO2-R7, and -COR7;

R7 is independently selected at each occurrence from the group:
alkyl of 1-6 carbons, aryl unsubstituted or substituted with 1-3 R8, and alkaryl of 1-10 carbons;

R8 is independently selected at each occurrence from the group:
OR9, NHR9, N(R9)2, CO2H, CO2R9, CONHR9, CON(R9)2, CN, COR9, CHO, SR9, SOR9, SO2R9, and NO2; and R9 is independently selected at each occurrence from the group:
H, alkyl of 1-6 carbons and aryl.

2. A compound of Claim 1 wherein:

X is a single bond, O, S, SO, SO2, CH2, CH2CH2, CH=CH, C=O, NR6;

a is a single or a double bond;

b is a single bond;

R1 is 2-, 3- or 4-pyridyl, or 4-pyrimidinyl;

R2 is -(CH2)m-W, wherein m = 1-4, and W is selected from the group:
2-, 3- or 4-pyridyl, 2-, 4-, or 5-pyrimidinyl, or 2-pyrazinyl; or R2 is -(CH2)n-Y, wherein n is 1 to 6, and Y is selected from the group:
CO2R7, CN, COR7, CHO, -OCOR7;

R3 and R4 are each independently selected from the group:
H, alkyl of 1-6 carbon atoms or -CH=CH-CH=CH-to form a fused ring; and R5 is selected from the group:
H, alkyl of 1-6 carbons, phenyl unsubstituted or substituted with 1-3 R8, F, Cl, Br, I, NO2 or -CH=CH-CH=CH- which attaches to an adjacent carbon forming a fused ring.

3. A compound of Claim 2 wherein:

X is O, S, SO, SO2, CH2, CH2CH2, C=O;

R2 is -(CH2)m-W, wherein m = 1, and W is selected from the group:
2-, 3- or 4-pyridyl, and 4-pyrimidinyl; or R2 is -(CH2)n-Y, where n = 3-4, and Y is selected from the group:
CO2R7, CN, and -OCOR7;

R3 and R4 are each H, or R3 and R4 join together forming a fused ring consisting of -CH=CH-CH=CH-; and R5 is selected from the group:
H, Cl, Br, I, NO2 or -CH=CH-CH=CH- which attaches to an adjacent carbon atom forming a fused ring.

4. The compounds of Claim 1 which are:

(a) 2,2-Bis(4-pyridinylmethyl)-pyrrolo[3,2,1-kl]phenothiazin-1(2H)-one;

(b) 1,1-Bis(4-pyridinylmethyl)-5,6-dihydro-4H-pyrollo[3,2,1-ij]quinolin-2(1H)-one;

(c) 2,2-Bis(4-pyridinylmethyl)-6,7-dihydro-indolo[1,7-ab][1]benzapin-1(2H)-one;

(d) 2,2-Bis(4-pyridinylmethyl)-pyrrolo[3.2.1-kl]phenoxazin-1(2H)-one;

(e) 2,3-Dihydro-6,6-bis(4-pyridinylmethyl)-pyrrolo[1,2,3-de]-1,4-benzothiazin-5(6H)-one;

(f) 1,2-Dihydro-2-(4-pyridinylmethyl)-2-(pentanenitrile)-pyrrolo[3.2.1-kl]phenoxazin-1(2H)-one, hydrobromide.

5. A pharmaceutical composition comprising a pharmaceutically suitable carrier and a therapeutically effective amount of a compound of Claim 1.

6. A pharmaceutical composition comprising a pharmaceutically suitable carrier and a therapeutically effective amount of a compound of Claim 2.

7. A pharmaceutical composition comprising a pharmaceutically suitable carrier and a therapeutically effective amount of a compound of Claim 3.

8. A pharmaceutical composition comprising a pharmaceutically suitable carrier and a therapeutically effective amount of a compound of Claim 4.

9. A method of treating a neurolgical disorder in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of Claim 1.

10. A method of treating a neurolgical disorder in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of Claim 2.

11. A method of treating a neurolgical disorder in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of Claim 3.

12. A method of treating a neurolgical disorder in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of Claim 4.