AU5279502A - Treatment of schizophrenia with ampakines and neuroleptics - Google Patents

Description

LO'J V. .L.LJ. ZAA U.L017 00~ OVV ita.rG vL.__ 7- 613 9663 3099 LWj U U 1

AUSTRALIA

Patents Act 1990 CORTEX PHARMACEUTICALS, INC.1 THE REGENTS OF THE UNIVERSITY OF CALIFORNIA COMPLETE SPECIFICATION STANDARD PATENT Invention Title:- Treatment of schizophrenia with ampakines and neuroleptics The following statement is a full description of this invention including the best method of performing it known to us:- RECEIVED TIME 28. JN. 11:07~ IM 8.JJ. 12 PRINT TIME 28. JUN. I I 2 8 '/UD UL 11:12 FAA 1i l9lU3 JUVU F.B. RKJI; CO. 007 613 9663 3099 1A Treatment of Schizophrenia with Ampakines and Neuroleptics The present invention claims priority from U.S. provisional application serial No.

60/063.627. filed on October 27. 1997. hereby incorporated by reference.

BACKGROUND OF THE INVENTION This invention relates to treatment of schizophrenia and other psychotic disorders. This invention especially relates to treatment of schizophrenia and other psychotic disorders by enhancement of receptor functioning in synapses in brain networks responsible for higher order behaviors. In particular, the invention provides methods for the use of AMPA receptor upmodulators in conjunction with antipsychotics for the treatment of schizophrenia.

The release of glutamate at synapses at many sites in mammalian forebrain stimulates two classes of postsynaptic receptors. These classes are usually referred to as a-amino-3hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/quisqualate and N-methyl-D-aspartic acid (NMIDA) rcceptors. AMPA/quisqualate receptors mediate a voltage-independent fast excitatory post-synaptic current (the fast epsc) whereas NMDA receptors generate a voltagedependent. slow excitatory current Studies carried out in slices of hippocampus or cortex indicate that the AMPA receptor-mediated fast epsc is by far the dominant component at most glutamatergic synapses under most circumstances.

AMPA receptors are not evenly distributed across the brain but instead are largely restricted to telencephalon and cerebellum. These receptors are found in high concentrations in the superficial layers of neocortex. in each of the major synaptic zones of hippocampus. and in the striatal complex, as reported by Monaghan et al.. in Brain Research 324:160-164 (1984).

Studies in animals and humans indicate that these structures organize complex perceptual-motor processes and provide the substrates for higher-order behaviors. Thus. AMPA receptors mediate transmission in those brain networks responsible for a host of cognitive activities.

RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28, JUN. 11:28 to/uu vz Jii:Jz rAA 013 U0o.) 3UV r.b. MI~JL L0. 1U U -613 9663 3099 Schizophrenia is a chronic disease that is characterized by positive (hallucinations.

delusions). negative (social wvithdrawal, flattened affect) and cognitive (formal thought, disorder.

executive memory dysfunction) symptoms. The dopamine hypothesis. that schizophrenia stems S from excessive midbrain dopamine transmission. originated from studies with rneuroleptics that revealed corrclations between clinical efficacy, effects on dopamine metaboli-sm (Carlsson Lindqvist. A4cia Pharmacol. Toxicol. 20:140-144, 1967) and binding to dopamine receptors (Creese ei al.. Science 192:481-482. 1976). In addition. drugs that increase synapt ic dopamnine concentration. amphetamines) produce aberrant sicreotyped behavior in animals (WT McKinney. in SC Shultz and CA Tarnrninga (eds) 3chizophrenia. Scientific Progr'ss. Oxford University Press. New York- pp 141-154. 1989) and schiztophrenia-like symptoms in humans (Snyder. .m .1.J Psycho!. 130:61-67. 1976).

However, accumulating evidence suggests that schizophrenia may also be caused by reduced neocorical sglutamatergic function, In vivo imaging studies have shown reduced metabolic activity (Andreasen ef al.. Lancet 349:1 730-1134. 1997: Weinbergcr and Berman.

PJhilos. Trauns. R Soc. Lond. B Biol. Sci 351:1495-1503- 1996) in frontal and temporal cortices that are rich in -lutamnateraic (excitatory) synapses. H-istopathologi tde aedcmne cytoarchitectural abnormalities (reviewed in Weinberger and Lipska. Schizophrenia Res, 16:87- 110, 1995). as well as reduced neuron or synapse densities and reduced AMA receptor (AMPA-R) densities in these same areas in post-mortemn schizophrenic brain (Easnwood etal..

BiaL PsYchiany 41:636-643. 1997). including hippocarnpus (Breese el al.. Brain Res. 674:82- 1995). This evidence is further supported by recent molecular studies That showed decreased AMPA-R subunit mRNA prevalence in neocortex (Eastwood et al.- AMcI. Brain Res. 29:211- 223. 1995) and hippocampus of schizophrenic brains (Eastwood et.aL Mol. Brain Res- 44:92- 98, 1997). Neurochemiucal studies have found reduced £tlutarrate concentrations In cerebrospinal fluid (Kim et al.. Neu.roscience Letrer 20:379-382. 1980) and lower glutamate and aspartaze levels in prefrontal and temporolirnbic areas (Tsai et al.. Arch- Gen. Psychiatry 52:829-836, 1995). Finally. phencyclidine (PCP), ketarnine and other use-dependent antagonists at NMDA- R produce aberrant behavior in animals (F'reed er al-, Ps-ychopharniacology 71: 29 1-297, 1980).

exacerbate symptoms in patients (Lahti et cit.. N\europs vchopha-mcicoloi:- 13:9.19- 1995). and produce a range of psychotic symptoms in volunteers That can accurately mimic Symptoms of schizophrenic patients (Krysial ef al.- .4rch, Gen. PsYchiatrn 51:199-214, 1994). T'hus.

RECEIVED TIME 28. JUN. I I: 07 PRINT TIME 28. JUN. 11I:2 8 La/uo uL 11;l: FrAA O.IJ ODJ JUV r.B. KJItA 1O. I_ UU 613 9663 3099 significant recent evidence supports the -hypofrontality' hypothesis of reduced excitatory tone in fronto-temporal cortices of the schizophrenic brain.

For the reasons set forth above, drugs that enhance the functioning of AMPA receptors have significant benefits for the treatment of schizophrenia. See. U.S. application Serial No. 08/521.022. Such drugs should also ameliorate the cognitive symptoms that are not addressed by currently-used antipsychotics. Experimental studies. such as those reported by Arai and Lynch. Brain Research. 598:173-184 (1992). indicate that increasing the size of AMPA receptor-mediated synaptic response(s) enhances the induction of long-term potentiation (LTP). LTP is a stable increase in the strength of synaptic contacts that follows repetitive physiological activity of a type known to occur in the brain during learning. Compounds that enhance the functioning of the AMPA form of glutamate receptors facilitate the induction of LTP and the acquisition of learned tasks as measured by a number of paradigms: Granger et al..

Synapse 15:326-329 (1993); Staubli et al., PNAS 91:777-781 (1994): Arai et al.. Brain Res.

638:343-346 (1994): Staubli et al.. PNAS 91:11158-11162 (1994): Shors et al.. Neurosci. Let.

186:153-156 (1995); Larson er al., J. Neurosci. 15:8023-8030 (1995); Granger er al., Synapse 22:332-337 (1996); Arai. et al.. JPET 278:627-638 (1996): Lynch et al.. Interna. Clin.

Psychopharm. 11:13-19 (1996); Lynch et al., Exp. Neurology 145:89-92 (1997): Ingvar et al., Exp. Neurology 146:553-559 (1997); and Lynch and Rogers. WO 94/02475 (PCT/US93/06916).

There is a considerable body of evidence showing that LTP is the substrate of memory.

For example, compounds that block LTP interfere with memory formation in animals. and certain drugs that disrupt learning in humans antagonize the stabilization of LTP, as reported by del Ccrro and Lynch. Neuroscience 49:1-6 (1992). A possible prototype for a compound that selectively facilitates the AMPA receptor was disclosed by Ito et al.. J. Physiol. 424:533-543 (1990). These authors found that the nootropic drug aniracetam (N-anisoyl-2-pyrrolidinone) increases currents mediated by brain AMPA receptors expressed in Xenopus oocvtes without affecting responses by y-aminobutyric acid (GABA), kainic acid or NMDA receptors.

Infusion of aniracetam into slices of hippocampus was also shown to substantially increase the size of fast synaptic potentials without altering resting membrane properties. It has since been confirmed that aniracetam enhances synaptic responses at several sites in hippocampus- and that it has no effect on NMDA-receptor mediated potentials- See. for example, Staubli et al.. in Psychobiology 18:377-381 (1990) and Xiao et al.. Hippocampus 1:373-380 (1991). Aniracetam has also been found to have an extremely rapid onset and washout, and can be applied RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28A UN, 11:28 /UO UZ 11:1 AA 01J VOOJ JUUU k KJALt IO0._ W U 613 9663 3099 4 repeatedly with no apparent lasting effects: these are valuable traits for behaviorally-relevant drugs. Unfortunately. the peripheral administration ofaniracetam is not likely to influence brain receptors. The drug works only at high concentrations mM) and Guenzi and Zanetti, J.

Chromatogr. 530:397-406 (1990) report that about 80% of the drug is convened to anisoyl- GABA following peripheral administration in humans. The metabolite. anisoyl-GABA. has been found to have only weak aniracetam-like effects.

SUMMARY OF THE INVENTION The present invention is based on the discovery that synaptic responses mediated by AMPA receptors are increased by administration of a novel class of compounds known as Ampakines. In particular, the present invention is based on the discovery that Ampakines are useful to treat Schizophrenia or Schizophreniform Disorder or Schizoaffective Disorder or Delusional Disorder or Brief Psychotic Disorder or Psychotic Disorder Due to a General Medical Condition or Psychotic Disorder Not Otherwise Specified. It is now apparent that compounds of the Ampakine family can interact with neuroleptics/antipsychotics in reversing behavior in animals in ways that predict success in treating subjects diagnosed as suffering from schizophrenia or related disorders. The interaction with the antipsychotics in these animal models of schizophrenia is not only additive, but surprisingly synergistic. Thus. schizophrenia is treatable by compounds that enhance glutamatergic neural transmission.

The present invention comprises methods, compositions and kits for treating schizophrenia in a subject in need thereof by up-modulating the stimulatory effect of natural ligands of a-anmino-3-hydroxy-5-methyl-isoxazole-4-propionic acid ("AMPA") receptors. A number of glutamatergic up-modulators may be used in the present invention: for example, 7chloro-3-methyl-3-4-dihydro-2H-l,2.4 benzothiadiazine S.S. dioxide.

In one embodiment, the invention comprises administering to a subject an effective amount of a compound having the following formula (with ring vertices numbered as shown): RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. 11:27 28/05 '02 11:13 FAX 613 9663 3099 F.B. RIV~E Co. il 1 613 9663 3099 R-=0

R

5 in' which in whO or 1

R

2 is a member selected from the group consisting of (CR8 2 )ii-m and Cn-mR 8 12(n-m)-2, in which n is 4, 5, 6, or 7, the W('s in any single compound being the same or different. each RO being a member selected from the group consisting of H and C I-C 6 alkyl, or one R 8 being combined with either R or R 7 to form a single bond linking the no. 3' ring vertex to either the no. 2 or the no. 6 ring vertices or a single divalent linking moiety linking the no. 3' ring vertex to either the no. 2 or the no. 6 ring vertices, the linking moiety bcing a member selected from the group consisting of CI1 2 CH=CH. 0. NH. N(CI-C 6 alkyl). N=CH.

N=C(Cr.'C 6 alkyl). CH(OH). and N(Ci-C( 6 alkvl

R

3 when not combined with any R8. is a member selected from the group consisting of C I-C6 alkyl. and C 1

-C

6 alkoxy;

R

4 is either combined with R, or is a member selected from the group consisting of H.

OH-. and Ci-CQ, alkoxy; is either combined with R.

4 or is a member sclccied from the group consisting of H4.

OH. C 1

-C

6 alkoxy, amino, mono(C 1

-C

6 alkyl)amino. di(C,-C 6 alkyl)amino. and

CH

2

OR

9 in which R! is a member selected from the group consisting of H.

C I-C 6 alkyl, an aromatic carbocyclic moiety, an aromatic heterocyclic moiety. an aromatic carbocyclic alkyl moiety. an aromatic heterocyclic alkyl moiety. and any such moie-cy substituted with one or more members selected from the group RECEIVED TIME 28. JUN, 11I: 07 PRINT TIME 28. JUN. 11I:2 7 Zb/UU '02 11:13 FAX 613 9663 3099 F.B. R]E.E Co. 613 9663 3099 10j012 6 consisting of C 1 alkyl. C 1

-C

3 alkoxv. hvdrox\y. lialo, amino. alkylarnino.

dialkylamirio. and methylenedioxy:.

is either H or CH4OR R' and R 5 when combined. form a member selected from the P-roup consisting of

RCR

1 1 N C qRz12 2q-1/ a/N "qR' 2 2 1 4 and 1 12 in which:

RW

0 is a member selected from the group consisting of 0. NH and N(C 1 -0 6 alkyl): R" is a member selected from the group consisting of 0. NH- and N(C 1

-C

6 alkyl): R 1 2 is a member selected from the group consisting of Hi and Cl-Cs alkyl. and when two or more R 1 2 ,S are present in a single compound. such R 12 's are the same or different.

p is 1. 2. or 3: and q is I or 2: and R. when not combined with any R 8 is a member selectcd from the group consisting of 14, C alkyl. and C I-C 6 alkoxy.

RECEIVED TIME 28. JN. 11:07 ITTME2.JJ.1:7 PRINT TIME 28AUN, 11:27 W v4 ±1x;xo~ rnA oxi voo ourr r b. £mJ&,L LJ 613 9663 3099 7 A preferred embodiment CX5 16. has the following-structure: 0 In another embodiment- the Ampakine has the followingt structure: Omn 0 N R 2 N R

I/N

in which.- R' is oxygen or sulfur:

R

2 and- W are independently selected from the group consisting of -CR~e. and

CX=:

M is =N or =CR 4 wherein R 4 and Re are independently R or top-ether form a single linking moiety linking M to the ring vertex the linking rnoiciy being2 selected from the group consisting of a single bond, -CR 2 -CR=CR-. and R. and R 7 are independently selected from the group consisting of 2 CR=CR-. -CR=CX-. -CX 2 and and

R

6 is selected from the group consisting of -(CR 2 -CR=CR-.

CR

2 and wherein X is -Br. -Cl. -CN. -OR. -SR. or -CONR 2 and R is hydrogen. C I-C, branched or unbranched alkyl. which mav be urisubstituted or substituted with one or more funCTionaliries defined above as X. or aryl. which may be unsubstituted or substituted with one or more functionalities defined above as X 253 m and p are independently 0 or 1, n and yare independently 0. 1 or 2.

RECEIVED TIME 28. JUN. 11:07PRN TIE2.J.1:2 PRINT TIME 28AUN. 11:27 28/06 '02 11:13 FAX 613 9663 3099 F.B. RIV~E Co. Z~014 613 9663 3099 8 Preferred cmbodjrnents include: 0 0N

N

00 0 N

NN

00

'N

ypca Apkie osgs orsytmi amiisrtin anrng fommllgrmst decarms erk- ei 0 S Typca Ampakine dosagens ofo stheicnto amnre t can amngseoe te from milg amst typical or atypical antipsychofic drug. Typical antipsychotics include: haloperidol. fluphenazine.

per-phenazine. chlorpromazine. rnolindone. pirnozide. trifluoperazine and thioridazine. and others- Atypical antipsychotics include: clozapine. risperidone. olanzapine. sertindole- M1 00907. ziprasidonc. seroquel. zotepine. arnisulpridc. iloperidone and others. The antipsvchotic drug may be administered at a subtherapeutic doses. at a lower dose than the dosage that is typically used for treaiments with the antipsychotic drugs alone.

Kits containing the compositions in the form of tablets or amnpules or other suitable packaging me-ans. formulaied for controlled dosage administration, are also provided.

RECEIVED TIME 28. JUN. I I: 07 PRINT TIME 28. JUN, 11I:2 7 -613 9663 3099 9 BRIEF DESCRIPTION OF THlE DRAWINGS Fias. 1 and 2 show that a representative Ampakine (CXS 16) synergistically enhanccs clozapine antagzonism of inethamphetaxnine. induced rcari ng activity.

In Figz. 1. behavioral activity was monitored with a computerized photobeamn system as described in Example 2. Each point represents the mean curnulative rearing score for the previous 10 minute interval. There was a large induction of rearing activity by mg/kg methamnphetarnine compared to saline vehicle. Clozapine (1.0 mg-p/kg,) had no significant effect on miethamphetatnine-induced stereotypic rearing. CX5 16 (10 i-ngikg) produced a small.

but statistically Insignificant aniagonism of methamphctamine-induced rearing. However.

together CX5 16 (10 mg/kg) and Clozapine (I mg/kg) produced a synergistic interaction.

reducine methamphetarnine-induced stereotypic rearing to a level nearly equivalent to that of v'ehicle-treated rats (nlo metharnphetarnine).

Fig. 2 provides a bar graph that shows total cumulative rearing activity during the 90 minute period after drug administration -Mean =standard error and number of animals for the experimental groups are as follows: saline. 56 9 n 12. MET14 (2 mg/kg). 724 136 n =_0 METH clozapine (1-0 mg/kg). 760OL 146 n =18: MET1H-; CX516 (10 mg/kg). 49.5' 78 n 19:. METH -CX316 clozapine- 125 22 n 17 p 0.0005 vs MEWI- clozapine (1 .0 m/g) by 2-tailed, unpaired rtrest assumning unequal variance: ttest uses mean sadr deviation).

RECEIVED TIME 28. JUN. 11:07PRN TIE 2.JN 1:1 PRINT TIME 28-JUN, 11:27 28/Ui 'UZ 11:14 FAX 613 9663 3099 F.B. R]ATE Co. 141016 -613 9663 3099 Figs. 3 and 4 show that a representative Aknpakine (CXS 16) synergistically nane haloperidol antagonism of rnethamphetarnine-induced stereotypic rearing activity.

Fig. 3 shows the antagonistic effect of CX516 (30 m2I/kag haloperidol (HAL:0.06 mg/kg) or 16 (30 mg/kg) combined with HAL (0.06 mg/kg) on rearing activity induced by methampheiamine. Neither CX5 16 (30 mg/kg) nor HAL (0.06 significantly reduced methamphetarnine-induced rearing activity (23% and 16%. respectively). However, the combination of those same doses was synergistic, reducinga merhamplietamine-induced rearing activitv by 67%.

Fig. 4 provides a bar graph that shows total rearing activity during the 90 minute test period.

Mean rearing activity t± standard error for the groups Is as follows: saline: 53 11I n z16- MFT!]: 105 16 1. n tz16: ]METH HAL (0.06 mg/kg): 934 119. n 16: METH CX516 13 (30 rne.,ka): 863 169.. n 16; METH HAL CX516: 360 77. n 16 0,0005 versus METH- HAL 0.06 rnsz/ke by 2-tailed, unpaired t test assuminga unequal v~ariance), DETAILED DESCRIPTION OF THE INVENTION The present invention is based on the discovery that synaptic responses mediated by AMPA rcceptors are increased by administration of a novel class of compounds knowNk as Arnpakines. disclosed in International Patent Application Publication No. WO 94/02475 (PcTr/u.S93/06916) (Lynch and Rogers. Regents of the University of California) and in related U.S. Patent No. 5,773.434. The invention is particularly based on the discovery that compounds of the .\jnpakine family can interact with antipsychotics/neuroleptics in reversing behavior in animals in ways that predict success in treating subjects diagnosed as schizophrernics.

Ampakines primarily act, not by directly stimulating neural activation. but by upmodulating ("allosteric modulation") neural activation and transmission in neurons that RECEIVED TIME 28. Jb. 11:07 PITTM 8 L1~ 12 PRINT TIME 28. JUN, I I 2 7 28/0o 'UZ 11:14 FAX 613 9663 3099 F.B. RI:E Co. 1 017 613 9663 3099 11 contain glutamatergic receptors. These compounds bind to the glutamate receptor at a site other than the glutamate binding site. but such binding does not by itself give rise to ion fluxes.

However, when a glutamate molecule binds to a glutamate receptor that has bound to it a glutamatergic compound of the invention, the subsequent ion flux is of much longer duration.

Thus. in the presence of the compounds used herein. postsynaptic neurons are activated by much lower concentrations of glutamate than postsynaptic neurons that do not contain bound compounds.

Applications contemplated for Ampakines include improving the performance of subjects with sensory-motor problems dependent upon brain networks utilizing AMPA receptors: improving the performance of subjects impaired in cognitive tasks dependent upon brain networks utilizing AMPA receptors: improving the performance of subjects with memory deficiencies: and the like. Additional applications contemplated for Ampakines include restoring biochemical and synaptic balance between brain networks where an imbalance occurs due to decreased AMPA receptor currents. Such therapeutic uses would include, but are not limited to. psychiatric and neurological disorders such as schizophrenia and clinical depression.

In addition to data from animal and human studies that show that Ampakines improve cognitive performance, other tests, to be described below, indicate that Ampakines may eliminate the cortical/striatal imbalance known to occur in schizophrenia and to do so in a synergistic manner when administered with either typical or atypical antipsychoticsineuroleptics.

The interaction with the antipsychotics in these animal models of schizophrenia is not only additive, but. surprisingly synergistic. These and other aspects and advantages of the invention will become apparent from the description that follows.

RECEIVED TIME 28. JUN, 11:07 PRINT TIME 28. JUN. 11:27 28/06 '02 11:14 FAX 613 9663 3099 F.B. R]:E Co.

613 9663 3099 12 S018 A. Definitions Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All references are incorporated by reference for all purposes. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described. For purposes of the present invention- the following terms are defined below.

The term schizophrenia in the claims encompasses Schizophrenia or Schizophreniform Disorder or Schizoaffective Disorder or Delusional Disorder or Brief Psychotic Disorder or Psychotic Disorder Due to a General Medical Condition or Psychotic Disorder Not Otherwise Specified. and the symptoms of these disorders, are in large par as defined in the Diagnostic and Statistical Manual of Mental Disorder. fourth edition (DSMIV). The sections of the DSMIV that relate to these disorders are hereby incorporated by reference.

"Cvano" refers to the group -CN.

"Halogen" or "halo" refers to fluorine, bromine. chlorine, and iodine atoms.

"Hydroxy" refers to the group -OH.

"Thiol" or "mercapto" refers to the group -SH.

"Sulfamoyl" refers to the -SO 2

NH

2 "Alkyl" refers to a cyclic, branched or straight chain, alkyl group of one to eight carbon atoms. The term "alkyl" includes reference to both substituted and unsubstituted alkyl groups.

This term is further exemplified by such groups as methyl. ethyl, n-propyl. i-propyl. n-buryl, RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28, JUN. 11:27 28/06 '02 11:15 FAX 613 9663 3099 F.B. RJAkE Co. l019 613 9663 3099 13 t-butvl. i-butvl (or 2-methyipropyl). cvclopropylmethyl. cyclohe.\yl. i-amyl. n-amyl. and hexyl.

Substituted alkyl refers to alkyl as just described including one or more functional groups such as arvi. acyl. halogen. hydroxyl. amido. amino. acylamino. acyloxy. alkoxy. cvano, nitro.

thioalkyL. mercapto and the like. These groups may be attached to any carbon atom of the lower alkyl moiety. "Lower alkyl" refers to CI-C 6 alkyl, with C,-C 4 alkyl more preferred. "Cyclic alkyl" includes both mono-cyclic alkyls, such as cyclohexyl, and bi-cyclic alkyls. such as f3.3.0)bicyclooctane and 1 )bicycloheptane. "Fluoroialkyl" refers to alkyl as just described.

wherein some or all of the hydrogens have been replaced wvith fluoric -CF 3 or -CF 2

CFA)

"Aryvl" or "Ar" refers to an aromatic substituent which may bc a single ring or multiple ings which are fused together. linked covalently. or linked to a common group such as an ethylene or methylene moiety. Thc aromatic ring(s) may contain a heteroatom. such as phenyl, naphthvl. biphenyl, diphenylmethyl, 2.-dipheiyl- I -ethyl. thienyl, pyridyl and quinoxalyl. The term "arvi" or "Ar" includes reference to both substituted and unsubstituted aryl groups. If substituted. the aryl group may be substituted with halogen atoms. or other groups such as hydroxy'. cvano. ritro. carboxyl. alkoxy, phenoxy, fluoroalkyl and the like. Additionally, the aryl group may be attached to other moieties at any position on the aryl radical vhich would otheinvisc be occupied by a hydrogen atom (such as 2-pyridyl. 3-pyridvl and 4-pyridyO).

270 The term "alkoxy" denotes the group where R is lower alkylI. substituted lower alkyl. aryl. substituted aryl. aralkyl or substituted aralkyl as defined below.

The term "acyl" denotes groups where R is alkyl. subs tituted alkyl. alkoxy. aryl, substituted aryl, amino and alkyithioL-

-I-

"Carbocyclic moiety' denotes a ring structure in which all ring vertices are carbon atoms. The term encompasses both single ring structures and fused ing struIctures. E~xamples of aromatic carbocyclic moieties are phenyl and naphthyl.

"T~rerocclic miety"denotes a ring~ structure in which one or more 6~vrie r atoms ot-her than carbon atoms. the remainder being carbon atoms. Examples of non-carbon RECEIVED TIME 28. JUN. I11: 07 PRINT TIME 28. JUN. I11:2 7 28/06 '02 11:15 FAX 613 9663 3099 F.B. R]AME Co. ___Zl020 _613 663 399 14 atoms are N. 0. and S. The term~ encompasses both single ring structures and fused ring structures. Examples of aromatic heterocyclic moieties are pyridyl. pyrazinyl. pyrimidinyl.

quinazoly I. isoquinazolyl. benzofuiyl. isobenzofuryl. benzothLofuryl. indoly I. and indolizinyl.

The termi "an-ino" denotes the group NRIR.. where R and R' may independently be hydrogen. lower alkyl. substituted lower alkyl. aryl, substituted aryl as defined below or acYl.

Thc term "amido" denotes the group -C(0)NRR'. where R and R'may independently be hvdroaen, lower alkyl. substituted lowver alkyl. aryl. substituted aryl as defined below.\ or acyl.

The term 'independently selected" is used herein to indicate that the two R groups. R, and R2. may be identical or different both R I and R 2 may be halogen or. R'may be halogen and R32 may be hydrogen. etc.).

The term "subject" means a miamrmal. particularlY a humian. The term specifically includes domestic and common laboratory marrmals. such as non-human primates. kine. horses.

pigs. goats- sheep. rabbits. rats and mice.

"a-amino--3-]ivdroxv,,-5-methivl-isoxazole-4-propionic acid". or "AMPA". or "lutarnatergi c" receptors are molecules or complexes of molecules present, in cells. particularly neurons. usually at their surface membrane. that recognize and bind to glutamate or AMPA_ The binding of AMPA or -lutamnate to an AMPA receptor normally give riet a -scries o f molecular events or reactions that result in a biological response. The biological response may be the activation or potentiation of a nervous imnpulse. chiange in celllar secretin o metabolism, or causinry cells to undergo differentiation or movement.

The term "central nervous system" or "CNS" comprises the brain and the spinal cord.

The term "peripheral nervous svstem' or "PNS" comrprises all parts of thie nervous systemn that are not part of the CNS_ including cranial and spinal nerves and the autonomic nervous system.

RECEIVED TIME 28A UN. 11 :07PRN TIE 2.J. 11 6 PRINT TIME 28AUN. 11:26 28/06 '02 11:15 FAX 613 9663 3099 F.B. RWEM Co. LOJ 021 -613 9663 3099 The phrase "effective amnount" means a dosage sufficient to produce a desired result.

Generally. th-e desired result is a subjective or objective decrease in the synmptoms of schizophrenia. as measured by the techniques described below.

B. Compounds used to treat schizophrenia Compounds usefu~l in the practice of this invention are g-enerally those which amplify (upmodulzte) dhe activity of the natural stimulators of AMPA receptors. particularly by amplifying excitatory synaptic response. We describe herein a wide variety of diverse compounds suitable for use in the invention. Methods for ideniI~ingp other compounds are routine. They involve a variety of accepted tests to determine wvhether a given candidate compound is an uprnodulator of the AMPA receptor. The primary assay is measurement of enlaraement of the excitatory postsvnaptie potential (EPSP) in i17 ira brain slices, such as rat hippocampal brain slices.

In expcriments of this kdid, slices of hippocampus from a mammal such as rat are prepared and maintained in an interface chamber using conventional methods. Field EPSPs are recorded in the stratum radiatum of region CAlb and elicited by single stimulation pulses delivered once per 20 seconds to a bipolar electrode positioned in the Schaffer-commissural projections (see Granger el cal., 1993. Synapse, 15:326-329: Staubli et al.. 1994a. Proc. Not.

Acad SO-. 9J:777-781; and Staubli, V. et al., 1994b, Proc. Nar. A4-ciL ScL 91:11158-11162; Arai er al.. 1994. Brain Res.. 638:343-346; Arai et al., "Effects of a centrally active drug on AMPA receptor kinetics).

RECEIVED TIME 28. JUN. 11 0Y PRINT TIME 28. JUN. 11I: 26 28/06 '02 11:15 FAX 613 9663 3099 F.B. R]A;E Co. 022 613 9663 3099 16 The wave form of a normal EPSP is composed of: an AMPA component. which has a relatively rapid rise time in the depolarizing direction (-5-10 msec) and which decays within -20 msec.; an NMDA component (slow -30-40 msec rise time and slow -40-70 msec decay) (the NMDA portion will not appear in normal CSF media, due to the voltage requirement for NMDA receptor channel activation, but in low magnesium media. an NMDA component may appear; a GABA component in the opposite (hyperpolarizing) direction as the glutamaiergic (AMPA and NMDA) components. exhibiting a time course with a rise time of -10-20 msec and very slow decay (-50-100 msec or more).

The different components can be separately measured to assay the effect of a putative AMPA receptor enhancing agent. This is accomplished by adding agents that block the unwanted components. so that the detectable responses are essentially only AMPA responses.

For example, to measure AMPA responses, an NMDA receptor blocker AP-5 or other NMDA blockers known in the art) and/or a GABA blocker picrotoxin or other GABA blockers known in the an) are added to the slice. To prevent epileptiform activity in the GABAblocked slices, known agents such as tetrodotoxin may be used.

AMPA upmodulators usefil in the present invention are substances that cause an increased ion flux through the AMPA receptor complex channels in response to glutamatergic stimulation. Increased ion flux is typically measured as one or more of the following nonlimiting parameters: at least a 10% increase in decay time. amplitude of the waveform and/or the area under the curve of the waveform and/or a decrease of at least 10% in rise time of the waveform, for example in preparations treated to block NMDA and GABA components. The increase or decrease is preferably at least 25-50%: most preferably it is at least 100%. How the increased ion flux is accomplished increased amplitude or increased decay time) is of secondary importance: upmodulation is reflective of increased ion fluxes through the AMPA channels, however achieved.

RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. 11:26 Zb/UU 'UZ 11:16 FAX 613 9663 3099 F.B. RJAUE Co.

613 9663 3099 I1I023 17 An additional and more detailed assay is that of excised patches. mcmbrane patches excised from cultured hippocampal slices: mnethods are described in Aral et al.. 1994. Outsideout patches are obtained from pyramidal hippocaxnpal neurons and transferred to a recording chamber. Glutarnate pulses are applied and data are collected with a patch clamp amplifier and 3 digitized (Arai et ali.. 1994).

Because these membrane patches should contain only glutarnateraic receptors.

GABAergtic cur-rents N~ill not be seen. Any NMDA currents can be blocked as above with The central action of a drugp can be verified by measurement of Field EPSPs In behaving animals (see Staubli ei al., 1994a) and time course of biodistribution can be ascertained via injection and PET measurement of radiolabeled drug (see Staubli ei 1994b), 4015 One such class of compounds is defined by Formula 1:

C=

2 In this formula: R1 is either N or CH: m is either 0or 1; R' is either (CR 8 1 -or ClR'(lI.in which; n is 4. 5- 6. or 7: and RECEIVED TIME 28. JUN. 11:07PRN TIE 2.J 1:6 PRINT TIME 28-JUN. 11:26 28/06 '02 11:16 FAX 613 9663 3099 F. B. RWAE Co. Fla024 613 9663 3099 18 the R 8 'S in any single compound are either the same or different- each R& beingy either H or C,-C 6 alkyl, or one Ra being combined with either R 4 or form a single band bridging the no. 3T and either the no. 2 or the no. 6 rinc, vertices or a sigl diaetlnigml ikng the no. 3' and either the no. 2 or the no. 6 ring vertices, examples of single divalent linking moieties being CH 2

CH,

2 -CH2. CH=CH-I 0. MiI. N(C 1

-C

6 alkyl), N=CH. N=C(C 1 alkyl), CH(OH), NHand N(C 1

-C

6 alkYl)-C(O); when not combined with any RK. is either H. C t-CA alkyl. or Cr-C 6 alkoxv R is either OH. or CI-C( 6 alkoxy, or is combined with R~: is either combined with R 4 or is OK. C 1 -Ci, alkoxy'. amino. n'iono(CI-C6 alkyflamino. di(C -C 6 alkyl)amino. or CH 2 OR in which: Rq is Fl C 1 -Cf alkyl. an aromatic carbocyclic moiety, an aromatic heterocyclic tnoietv. an aromatic carbocyclic alkyl moiety, an aromatic heterocyclic alkyl moiety, or any such moiety substituted with one or more members of the group C 1

-C

3 alkyl.. C 1 alkoxv. hydroxy. halo, amino, alkylamino. dialkylamino. and methivlertedioxy: R6' is either H or CI-bOR?,;

R

4 and R-5 when combined form a member selected from the group consisting of R11 R 11 (C2 p 7, (Cq R2q_1) 8.

N

R

8 %(CqR 12 /1112 R q q-1) 9 and R 1 in which:

R'

0 is either 0. NH- or N(Ci-Cf, alkyl): R" is either 0. NH- or N(C I-C 6 alkyl):- RECEIVED TIME 28. JUN. I11: 07 PRINT TIME 28. JUN. I11:26 '02 11:16 FAX 613 9663 3099 F.B. RAT~ Co. 4025 613 9663 3099 19 R1 2 is either H- or C,-C 6 alkyl,. and when two or more R 1 2 S are present in a single compound. such R' 2 's are the samne or different: p is 1. 2, or3;, and q is I or 2; and R 7 when not combined with any RB. is either H, C alkyl. or C I-Cs alkoxy.

Within the scope of Formula 1, certain subclasses are preferred. One of these is the subclass in which R 2 is (CHR 8 Or Cn,.HR 8 2 (,nm4.

3 and R 3 is H. Ci-CQ, alic I. or C 1

-C

6 alkoxy.

Another is the subclass in which R 2 is(CHIR),- or CFHRflun..-j, and one R 8 is combined with either R 3 or R 7 Lo form a sing-le bond bridging the 2 and 3' ring vertices or a single divalent linking moiety linking the 2 and 3'ring vertices. with CH 2

CH

2

-CH

2 CH=CH, 0. NH. N(C 1

-C

6 alkyl). N=CH. N=C(C 1

-C

6 alkyl). CH-(O1-D. NH-C(O). and N(C 1

-C

6 alkyvl)-C(O) as the linking moiety. A preferred subclass of R2 IS CHR 8 '-CI-l 2 -CI-1-CH 2 and

CHR-CH

2

-CH

2

,-CH-CH-

2 A preferred subclass of linking moieties is CH 2

)-CH

2 CH4=CH, 0, NH. and CH(OH). A further preferred subclass is CH,, 0. NH. and CH-(OH).

When R 4 and R 5 are combined. a preferred subclass for R 12 is H- and CH,, and preferred groups representing the combination of R 4 and are (C 1 2 1

I/

I- I-and R 12.

In these groups. R1 0 and R" are both preferably 0, and p is I or 2. Still further preferred subclasses are those in which mn is zero.

RECEIVED TIME 28-JUN. 11:07 RECIVD IM 28 JN.110~PRINT TIME 28. JUN, 11I: 26 28/06 '02 11:16 FAX 613 9663 3099 F.B. RWAE Co. -613 9663 3099 101026 A further class of compounds useful in the practice of the invention are those of Formula, H 2 NS02' 122 In Formula 11: is either 14,. halo or CF.., R22 and R 3 either are both H or are combined to form a double bond bridging the 3 and 4 ring vertices;

R

24 is either H. C 1 -Cs alkyl, CS-C, cycloalkyl. C- 5 -C7 cycloalkenyl, Ph. CH 2 Ph.

CHSCH

2 ,Ph, CHX., CHX,, CH 2

SCHCF

3

CH,SCH

2

?CH-CH

2 or C Hf~b14; and e~ is a member selected from the group consisting of H and C I-C 6 alkyl.

WVithin the scope of Formula 11. certain subclasses are preferred. One of these is the subclass in which R 2 1 is Cl or CF 3 with Cl preferred. Another is the subclass in which all X's are Cl. Still another is the subclass in which R 2 and RP are both A preferred subclass of R 24 is that which includes CH2Ph. CH-iSCH,-Ph. and C Preferred compounds within the scope of Formula 11 are those in which IZ24 is either C-i-C7 cycloalkyl. Cs;-C 7 cycloalkenyl or Ph denotes a phenyl group). other preferred compounds of this group are those in which R 2 is halo. R 22 is H. R23 is H. and R 2 5 is H.

Prefer-red substituents for R 24 are cyclohexyl, cyclohexenyl. and phenyll.

RECEIVED TIME 28. UN. 11:07PRN TIE 2.J. 11 6 PRINT TIME 28AUN. 11:26 Z?/U U UZ 11:17 FAA 51J 9t$UJ JU99 F.B. RJAE (SO. 613 9663 3099 LO 027 21 Compounds 1 through 25 below are examples of compounds within the scope of Formula I: 00r 0 00 0 2 00q

N

0 0-j 7 0ND bi0* 3 00r 0

CH

3

N

0 6

N

9 00 00 0 NN N NH N3 N J 12 13 14 ,1 5 3C CH RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28, JUN. I I 2 go/vo uz 11:11 VAA UIJ VUUJ JUVV F. B. KJA;E 613 9663 3099 I10 028 0 1 00N CH OCH 3 19 17 aniracetam

CH

3 0CH 2 ZCHZ0CZH 5 21

LH

2 0H 22 0 00

CH

2 -0 0 23 :H 20cH 3 24 RECEIVED TIME 28. JUN. 11:07 RN IE 2.JN 12 PRINT TIME 28, JUN, I I 2 6 ZO/vu Uz 11:17 -AJL U13 J J~U99 F.B. RIVE 613 9663 3099 23 Compounds 26 through 40 below are compunds within the scope of Formula 11: 0 0

H

2 NSO 2 S NH 26 bendraflumethiaZide H2 NSO2cS.N C1:: N K CH 2

SCH

2 27 benzthiazide 10j029 C1 ~N iCH 2

CH(CH

3

H

28 buthiazide 0 N 0 29 chlorothiazide C.NSO Icle N ICH 2

SCI{

2 CF 3 epithiazide cyclothiazide RECEIVED TIME 28-JUN, 11:07 RECIVD IM 28 JN. 1107PRINT TIME 28. JUN. II: 2 613 9663 3099 24 Wj U -j 'J 0 32 hydrochlorothiazide 33 hydroflumethiazide 0 /0 H2NSO2a- Sl CH 3 ci l: Nl CH 2

CI

H

34 methyiclothiazide s /CH 3 cl N CH 2

SCH

2

CF

3

H

36 polythiazide

H

37 trichlormethiazide cl N CHZSCI-I 2 CH= CH 2 methaithiazide 38

H

RECEIVED TIME 28-JUN. 11:07 REEIEDTIE 8.J~ CPRINT TIME 28. JUN. 11: 2 ~.~IUU xi±:ii raA 01.3 woi .3uv 11.b. KJAIL i UJJI 613 9663 3099 A particularly preferred compound is compound CX5 16. I -(Quinoxalin-6ylcarbonvl)piperidine. having the following struicture: 0 In another embodiment, the Arnpakine is a compound of formula III: omn 0 1 2 NR N~

R

\N RR W R in which: RW is oxygen or sulfur,

R

2 and RW are independently selected from tile group consisting of and

CX;=:

M is =N or e=CR-. wherein R 4 and R 8 are independently R or together form a single linking moiety linking M to the ring vertex the linking moiety being selected from the group consisting of a single bond. -CR 2 -CR=CR-, and R i and R 7 are independently selected from the group consisting Of -(C 2 CR=CR-. -CR=CX-. and and

R

6 is selected from the group consisting of -(CR 2 -CRCR, and wherein X is -Br, -Cl. -CN, -NO 2 -OR, -SR. -CQ 2 R. or -CONR 2 and RECEIVED TIME 28. UN. i, RN IE 2.JN 11:25 PRINT TIME 28AUN. 11:25 ilAM 513 WM3S. JU91 F.B. HJAkh CO.

613 9663 3099 ij U32 26 R is hydrogen. C I-C 6 branched or unbranched alkyl, which may be unsubstituted or substituted with one or more functionalities defined above as K. or aryl. which may be unsubstituted or substituted with one or more functionali ties defined above as X;, m and p are independently 0 or 1; n and y are independently 0. 1 or 2.

Preferred embodiments include- 0 N

C

RECEIVED TIME 28. JUN. 11:07 RN IE2.JN 12 PRINT TIME 28, JUN. I I 2 VW .II FZAA OVU. O uO F x~js,L M~ 613 9663 3099 27 1. Preparation of Formula I compounds The compounds described above are prepared by conventional methods know.\n to those skilled in the art of synth1etic organic chemistry. For example. certain compounds of Formula I are prepared from an appropriately substituted benizoic acid by contacting the acid under conditions suitable to activate the carboxy group for ihe formation of an amnide. This is accomplished, for example. by activating the acid with carbonyi diimidazole. or with a chlorinating ag-ent such as thionyl chloride or oxalyl chloride to obtain the corresponding bcn~zol chloride. The activated acid is then contacted wvith a nitro cen-containing heterocyclic compound under conditions suitable for producing the dcsired imide or amide. Alternatively, 1(1 the substituted benzoic acid is ionized by contact with at least tw,,-o equivalents of base such as triethylaminc in an inert solvent such as rnethylene chloride or alcohol-free chloroform. and the ionized bcnzoic acid can then be reacted with pivaloyi chloride or a reactiv'e carboxylic acid anhydride such as tifluoroacetic an~hvdride or trichloroacetic anhydride. to produce a mixed anhvdride. The mixed anhydride is then contacted with a nitrogen-containing heterocyclic compound to produce the desired imide or amide.

A~ further alterniative to these methods. suitable for some of the compounds of Formula 1.

is to contact thc appropriately selected 3.4-(alkylenedihetero)-benzaidehvde with ammonia to form an imine. thcni contacting the imine with benzoyloxycarbonyl chloride to form the benzoyloxvcarbonv I imine. Suitable 3 .4-(alkvlenedihetero)-benzaldehvdes include 3.4- (r'netlh I cned iox\ )-berizadehydc. 3 .4-(ethylenedioxy,)-benzaldellyde. 3 .4-(propylenedioxy)benzaldchvdc. 3 .4-(cthyl idenedioxy )-benzaldehvde. 3 A4-(propyienedEihio)-benzaldehy.de. 3,A- (ethyl Iidenedi thio)-benzaldehvde. 5-benzimidazolecarboxaldehvNde. and 6quinoxalindcarhoxaidehyde. The benzoyloxycarbonvl- imnine is then contacted with a simple conjugated diene such as butadiene under cvcloaddition reaction conditions, and then with a Lewis zicid under conditions suitable for a Friedel-Crafis acylation. Examples of suitable conjugated dienes include butadiene. 1.3-pentadiene. and isoprene. and examples of suitable Lewis acids include AIC1 3 and ZnCl 2 Still further compounds within Formula I are prepared from 2.3-dihvdrox\v naphthalene.

This staring material is reacted wvith 1.2-dibromoethane inl the presence of base to produce an cthylenedioxy derivative of naphthalene. which is then reacted with can oxidizina acent such as RECEIVED TIME 28. UN. 11:07PRN TIE 2.JN 1:2 PRINT TIME 28. JUN, I I 2 16V/ V V16 .LJL xo A u±x3 vuu.) o)Uv Zrf. £njs.L aL Wi- u'I'a 613 9663 3099 28 potassium permnanganate to produce 4.5-ethylenedioxvphthaldehydic acid. Thc latter is contacted with anhydrous ammonia to form an imine. which is then trated with a suitable carbonv I-activating a-cent such as dicyclohexylcarbodiirnide under cyclization conditions to form an acyl imine. The acyl imine is then reacted with a simple conjugated diene to achieve cycloaddition.

Still further compounds within Formula I are prepared by contacting- an a-lialotoluic acid with at least two equivalents of an alkali salt of a lower alcohol according to the Williamnson ether synthesis to produce an ether linklage. The resulting alkoxymethylbenzoic acid is activated with carbonvldiimidazole. thionyl chloride. dicyclohiexvlcarbodiiniide. or any other suitable activatingi aenT- and reacted with a suitable amnine to achieve a carboxamide linkace.

In an alternate to the scheme of the preceding paragrph afry-bsItedaoti carboxamide is prepared by activation of an appropriate starting acid with a tertiary amine: (for example. tricthvl amine) plus an acid chloride (for example. pivaloyl chloride) to produce a mixed andrinde for coupling to a suitable amine. The formyl gYroup is then reduced to an alcohol by a suitable reducing agent such as sodium borohydride. The alcohol is then converted to a leav'ing group which is replaceable by the alkali salt of an alcohol. The leaving group can be generated by reagents such as thionyl chloride. thionyl bromide. mineral acids such as hydrochloric. hydrobromic or hydroiodic acids, or the combined action of~ a tertiary amine plus either a suitable sulfonic anhvdride or sulfonyi halide. Alternatively, the alcohol is activated by removinge the proton. Tis is achieved by the action of a strong base such as sodium hydride in an aprotic solvent such as dimethylforrnaride. The resulting alkoxide is then reacted with a suitable lklhalide or other alkyl compound with a suitable leaving group to produce the desired ether linkage.

Fused ring structures such as those in which R5 and one of the R8's of Formula I are combined to form a single linking group bridging the 2 and 3 carbon atoms can be snthesized in the following manner. The carboxyl group of an appropriately substituted salicylic acid is 'O activated wvith carbonyldiimidazole in dichloromethane- chloroformn. tetrahydrofiuran- or other anhvdrous solvent. An aminoalkylaccial such as H 2

N(CH

2 )3CH(OCH2CH4) is then added. The resultinu amide is treated with an aryl or alkyl sulfonic acid. trifluoroacetic acid- or other strong RECEIVED TIME 28. UN. 11:07PRN TIE 2.J 12 PRINT TIME 28AUN. 11:25 613 9663 3099 29 acid. in a solvent of low basicity such as chloroform or dichloromethanc. to cleave the acetal and cyclize the intermediate aldehyde with the amide nitrogen and the phenolic oxygen.

In all of these reaction schemes, the methods and reaction conditions for each of the individual reactions are well within the routine skill of. and will be readily apparent to, the synthesis chemist.

2. Preparation of Formula II compounds Compounds of Formula II and methods for their preparation are described in the literature. These methods are within the routine skill of the synthesis chemist. The preparation of compounds such as bendroflumethiazide, for example. is described by Goldberg (Squibb). in U.S. Patent No. 3.265.573 (19661. The preparation of compounds such as benzthiazide, epithiazide. methalthiazide and polyhiazide is described by McManus (Pfizer). U.S. Patent No.

3.009.911 (1961). The preparation of buthiazide is described in U.K. Patent Nos. 861.367 and 885.078 (Ciba. 1961). The preparation of chlorothiazide is described by Hinkley (Merck U.S. Patent Nos. 2-809,194 (1957) and 2,937.169 (1960). The preparation of hydrochlorothiazide is described by Novello (Merck U.S. Patent No. 3.025.292 (1962); de Stevens and Wemer (Ciba), U.S. Patent No. 3.163.645 (1964); and Irons er al. (Merck U.S. Patent No. 3.164.588 (1965). The preparation of hydroflumethiazide is described by Lund at al. (L6vcns), U.S. Patent No. 3.254.076 (1966). The preparation of methylclothiazide is described by Close ea al., 1960. J. Am. Chem. Soc.. 82:1132, The preparation of trichlormethiazide is described by de Stevens et al., 1960, Experientia. 16:113. The disclosures of each of these patents and papers is incorporated herein by reference.

RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28, JUN. I I 2 nna V~u 00 r, J;1. &lJ*,L 9 W_ 613 9663 3099 3. screening of compounds A number of compounds belonging to the above-described genus have been shown to up-modulate glutamatergic transmission by augmenting ligand-AMPA receptor complexactivated ion gating. Staubli. U. et aL, 1 994a. Proc. Xat. Acad Sc. 9J :777-78 1: Staubli.

U. ei at.. 1994b, Proc. Nar. Acad Scd. US.A.. 91:1115sg-11162: Arai. A. et al.. 1994- Brain Res..

638:343-346: Granger. R. et oL. 1 993- Synapse. 15:326-329: all of which arc incorporated by reference. These compounds rapidly cross the blood-brain barrer (Staubli. U. ei at. 1 I994b) and increase ESPSs in freely moving rats (Staubli. U. et al.. 1994a). Animal experiments indicate that these centrally acTive modulators improve memory in both rat (Granger, R. et at., 1993; Staubli. U. et at.. 1994a) and human models (Lynch ei 1996. Internar. Clinical PsychophOY-rnaC0O.E' 11:13; Inavar et at.. 1997. Exp. Nkezrol. 146: 553-559. both of which are incorporated by reference).

Once prepared. the compounds of this invention are screened for their ability to amplify aq (upmodulate) the activity of the natural stimulators of AMPA receptors. particularly by amplify'ing excitatory synaptic responses. A variety of accepted tests are used to determine whether a cTi yen compound is an upmodulator of the AMPA receptor. The primary assay is measurement of the enlargement of the excitatory postsynaptic potential (EPS?) in in vitro brain slices. such as rat hippocarnpal brain slices.

In experiments of this kind, slices of hippocampus from a mammal. such as rat. are prepared and maintained in an interface chamber using conventional methods. Field EPSPs are recorded in the stratum radiatum of region CAlb and elicited by single stimulation pulses delivered once per 20 seconds to a bipolar electrode positioned in the Schaffer-commissural projections (see, Granger. R. el aL, Synapse. 15:326-329 1993. Staubli. U. et al.. 1994a. Proc.

Nai. .4cad Sci., 91:777-781. and Staubli. V. el 1994b. Proc. Vat. A cad Sci. 91:1 1158- 11162: Arai. A. ei at.. 1994. Brain Res., 638-343-346; Arai. A. et at.. "Effects of a centrally active drug on AMPA receptor kinetics, submitted). The wave form of a normal EPS? is ,0 composed of an AMPA component. which has a relatively rapid rise time in the depolarizing direction 10 msec) and which decays within -20 msec.:an NMDA component (slow -30-40 rnsec rise time and slow -40-70 msec decay) (the NMDA portion will not appear in normal or RECEIVED TIME 28. JN. 11:07PR TTIE 2.J. 1:2 PRINT TIME 28AUN, 11:25 613 9663 3099 31 artificial CSF (cerebro-spinal fluid) media- due to the voltage requirement for NMDA receptor channel activation, but in low magnesium media, an NMDA component may appear: a GABA (gamma-aminobutyric acid) component in the opposite (hyperpolarizing) direction as the glutamatergic (AMPA and NMDA) components. exhibiting a time course with a rise time of -10-20 msec and very slow decay (-50-100 msec or more).

The different components are separately measured to assay the effect of a putative AMPA receptor enhancing agent. This is accomplished by adding agents that block the S unwanted components, so that the detectable responses are essentially only AMPA responses.

For example, to measure AMPA responses, an NMDA receptor blocker AP-5 or other NMDA blockers known in the art) and/or a GABA blocker picrotoxin or other GABA blockers known in the an) are added to the slice. To prevent epileptiform activity in the GABAblocked slices, known agents such as tetrodotoxin may be used.

AMPA upmodulators useful in the present invention are substances that cause an increased ion flux through the AMPA receptor complex channels in response to glutamatergic stimulation. Increased ion flux is typically measured as one or more of the following nonlimiting parameters: at least a 10% increase in decay time. amplitude of the waveform and/or the area under the curve of the waveform and/or a decrease of at least 10% in rise time of the waveform, for example in preparations treated to block NMDA and GABA components. The increase or decrease is preferably at least 25-50%; most preferably it is at least 100%. How the increased ion flux is accomplished increased amplitude or increased decay time) is of secondary importance: upmodulation is reflective of increased ion fluxes through the AMPA channels. however achieved.

An additional and more detailed assay is that of excised patches, membrane patches excised from cultured hippocampal slices: methods are described in Arai et al., 1994. Outsideout patches are obtained from pyramidal hippocampal neurons and transferred to a recording chamber. Glutamate pulses are applied and data are collected with a patch clamp amplifier and digitized (Arai ct al.. 1994). Because no GABA is applied to the patch. GABAergic currents will not be elicited. Any NMDA currents are blocked as above with RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. I I: 2 -613 9663 3099 The central action of a drug is verified by measurement of field EPSPs in behaving animals ksee. Staubli ei aL., 1994a) and time course of biodistribution can be ascertained via injection and subsequent quantitation of drug levels in various tissue samples. Quantitation is accomplished by methods known to those skilled in the art and will vary depending on the chemical nature of the drua.

C. Other Compounds The above described genuis and species of compounds represent merely one example of iglutarnacergic compounds- that may be used to treat schizophrenia according to the present invention. The treatments provided by present invention are not limited to the compounds described above. The present invention also encompasses administering other compounds that enhance the stimulation of a-amino-S)-hydroxv-5;-methyl-isoxazole-4-proponic acid ("AMPA") receptors in a subject. said enhancement being sufficient to diminish the symptoms of schizophrenia. Examples of other such AMPA-selecti'e compounds include 7-chloro-3-methyl- 3-4-dihvd-'-21.. benzothiadiazine S.S. dioxide, as described in Zivkovic Ct aL. 1995, Phar-n~aeoI. Exvp. 7iiarap.. 272:300-309: Thompson et aL. 1995. Proc. Ala. Acad Sci. USA.

92:7667-7671.

The methods of the present invention also involve the administration of antipsychotic medications. Antipsychotic medications (this term is used interchangeably %%ith the termn neuroleptics) are a class of compounds that include haloperidol and at-ypical members such as clozapine. olanzapine and risperidone. Chiodo et aL. (1983) "Typical and atypical neurolepties: effects of chronic administration on the activity of A-9 and A-10 midbrain doparninergic neurons," J Neurosci. 3:1607-1609; Ljungberg ei &l (1978). "~Classification of neuroleptic drugs according to their ability to inhibit apomorphine-induced locomotion and gnwn:eidence for two different mecahnisms of action." PsychopharmacoloD 56:239-247; Lynch ei at.. (1988). Sensitization of chronic neuroleptic behavioral effects. 11O1. Ps ychiat-y.

24:950-951: Rupniak ez al- (1985). "Mesolimbic dopamnine Function is not altered during continuous chronic treatment of rats with typical or atypical neuroleptic drugs." J Neural.

Transm. 62-249-266:. Sayers ei ali. (1975), -Neuroleptic-induced hypersensitivity of striatal RECEIVED TIME 28. JUN. 11:07PITTM 2.JN 12 PRINT TIME 28AUN, 11:25 613 9663 3099 33 dopamine receptors in the rat as a model of tardive dyskinesia. Effects of clozapine. haloperidol.

loxapine and chloropromazine.' Psychopharmacologia 41:97-104: Titeler et al.. (1980), "Radioreceptor labeling of pre- and post-synaptic receptors." In Canabeni et al. (eds), "Long-term effects of neuroleptics." Raven Press. New York. Adv. Biochem. Psychopharmacol.

24:159166: Wyatt. R.J. (1976), Biochemistry and schizophrenia (par IV): the neuroleptics their mechanism of action: A review of the biochemical literature. P vchopharmacol. Bull.

12:5-50.

D. Subject selection Subjects contemplated for treatment in accordance with this invention include humans.

laboratory animals, and domestic animals. In particular. human subjects are individuals that exhibit symptoms of Schizophrenia or Schizophreniform Disorder or Schizoaffective Disorder or Delusional Disorder or Brief Psychotic Disorder or Psychotic Disorder Due to a General Medical Condition or Psychotic Disorder Not Otherwise Specified, as defined in the Diagnostic and Statistical Manual of Mental Disorder, third edition (DSMIV).

E. Administration of compounds The compounds of this invention are incorporated into a variety of formulations for therapeutic administration. Examples are capsules, tablets. syrups, suppositories, and various injectable forms. Administration of the compounds is achieved in various ways, including oral, bucal. rectal. parenteral_ intraperitoneal. intradermal. transdermal- eic.. administration. Preferred formulations of the compounds are oral preparations, particularly capsules or tablets.

RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. I I 2 613 9663 3099 34 F. Dosage The above described compounds and/or compositions are administered at a dosage that diminishes the symptoms of schizophrenia and related disorders. (see above) in subjects suffering from these disorders. while at the same time minimizing any side-effects. It is contemplated that the composition will be obtained and used under the guidance of a physician.

Typical dosages for systemic Ampakine administration range from about 0.1 to about 1000 milligrams per kg weight of subject per administration. A typical dosage may be one 500 mg tablet taken once a day. or one time-release capsule or tablet taken once a day and containing a proportionally higher content of active ingredient. The time-release effect may be obtained by capsule materials that dissolve at different pH values, by capsules that release slowly by osmotic pressure. or by any other known means of controlled release.

Dose levels can vary as a function of the specific compound, the severity of the symptoms, and the susceptibility of the subject to side effects. Some of the specific compounds that stimulate glutamatergic receptors are more potent than others. Preferred dosages for a given compound are readily determinable by those of skill in the an by a variety of means. A preferred means is to measure the physiological potency of a given compound that is a candidate for administration, by the method of Davis et al. (1996). submitted to Behavioral Neuroscience.

Briefly, excised patches and excitatory synaptic responses are measured in the presence of different concentrations of test compounds. and the differences in dosage response potency are recorded and compared. Davis er al. found that one specific compound designated BDP-20 was about ten-fold more potent than another designated BDP-12 in a variety of behavioral (exploratory activity, speed of performance) and physical (excised patches and excitatory synaptic responses) tests. The relative physiological potency was an accurate measure of their behavioral potency. Thus, excised patches and excitatory synaptic responses may be used to gauge the relative physiological (and behavioral) potency of a given compound with regard to a known standard.

RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. I I 2 613 9663 3099 Glutamatergic compounds for the treatment of schizophrenia may have a half-life measured from less than 10 minutes to more than 2 hours. In some embodiments. the compound preferably has a rapid onset and short elimination half-life (S 90 min.).

In the present invention, the Ampakines are typically administered together with antipsychotic compounds. Although the antipsychotic drugs are effective in their normal therapeutic range compounds are preferably adminsiercd close to or at subtherapeutic doses, i.e., doses lower than the doses typically used for administration of the antypsycholic by itself to treat disorders such as schizophrenia. See, e.g. U.S. Patent No. 5.602.150. The range of therapeutically effective doses for mammalian subjects may range from about 0.1 to about 2000 mg per kilogram of body weight per day, or preferably between about 1 mg/kg to about 500 mg/kg of body weight per day. more preferably between about 10 mgikg to about 250 mg/kg.

depending on the particular neuroleptic administered, route of administration, dosage schedule and form. and general and specific responses to the drug. For convenience, the total daily dosage may be divided and administered in portions throughout the day. if desired. The therapeutically effective dose of antipsychotic drugs administered to adult human patients also depends on the route of administration, the age, weight and condition of the individual. Some patients who fail to respond to one drug may respond to another, and for this reason, several drugs may have to be tried to find the one most effective for an individual patient. Some therapeutic doses are shown below:: ANTIPSYCHOTIC SUGGESTED THERAPEUTIC DOSAGE RANGE (mg/kg body weight Chlorpromazine (Thorazine) 100-1000 Thioridazine (Mellail) 100-800 Mesoridazine (Lidanar. Serentil) 50-400 Piperacetazine (Quide) 20-160 Trifluoperazine (Stelazine) 5-60 Perphenazine (Trilafon) 8-64 Fluphenazine (Permitil. Prolixin) 2-20 Thiothixene (Navane) 2-120 RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28A UN, 11:24 613 9663 3099 36 Haloperidol (Haldol 2-20 Loxapine (Loxitane) 20-160 Molindone (Lidone. Moban) 20-200 Clozapine (Clozaril) 25-400

EXAMPLES

The following examples are submitted for illustrative purposes only and should not be interpreted as limiting the invention in any way. A person of ordinary skill, with knowledge of this invention and of the prior art. will readily think of other subjects. other dysfunctions. and other glutamatergic substances that are readily substituted in the following examples. Also, the patents and publications cited in this disclosure reflect the level of skill the art to which this invention pertains, and are herein individually incorporated by reference to the extent that they supplement, explain, provide a background for or teach methodology, techniques and/or compositions employed herein. Those of skill in the art will readily appreciate that the foregoing protocol can be used, with only minor modifications, to prepare the other compounds of the present invention.

Example 1 In Vitro Phvsiological Testin The physiological effects of Ampakines may be tested in virro with slices of rat hippocampus according to the following procedure. Excitatory responses (field EPSPs) are measured in hippocampal slices, which are maintained in a recording chamber continuously perfused with artificial cerebrospinal fluid (ACSF). During a 15 30 minute interval. the perfusion medium is switched to one containing various concentrations of the test compounds.

Responses collected immediately before and at the end of drug perfusion were superimposed in order to calculate both the percent increase in EPSP amplitude and percent increase in the width of the response at one-half the peak height (half-width).

RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN, I I 2 613 9663 3099 37 To conduct these tests. the hippocampus was removed from anesthetized. 2 month old Sprague-Dawley rats and in vitro slices (400 micrometers thick) were prepared and maintained in an interface chamber at 35 "C using conventional techniques [see. for example, Dunwiddie and Lynch. J. Physiol. 276: 353-367 (1978)]. The chamber was constantly perfused at 0.5 mL/min with ACSF containing (in mM): NaCI 124. KCI 3. KH2PO4 1.25. MgSO4 CaCl1 3.4. NaHCO 3 26, glucose 10 and L-ascorbate 2. A bipolar nichrome stimulating electrode was positioned in the dendritic layer (stratum radiatum) of the hippocampal field CA1 S close to the border of field CA3.

Current pulses (0.1 msec) through the stimulating electrode activate a population of the Schaffer-commissural (SC) fibers which arise from neurons in the field CA3 and terminate in synapses on the dendrites of CAI neurons. Activation of these synapses causes them to release the transmitter alutamate. Glutamate binds to the post-synaptic AMPA receptors which then transiently open an associated ion channel and permit a sodium current to enter the postsynaptic cell. This current results in a voltage in the extracellular space (the field excitatory post-synaptic potential or field "EPSP") which is recorded by a high impedance recording electrode positioned in the middle of the stratum radiatum of CAl.

For experiments designed to test the ability of compounds to enhance AMPA receptor currents- the intensity of the stimulation current was adjusted to produce half-maximal EPSPs (typically about 1.5 2.0 mV). Paired stimulation pulses were given every 40 sec with an interpulse interval of 200 msec (see below). The field EPSPs of the second response were digitized and analyzed to determine amplitude, half-width, and response area. If the responses were stable for 15-30 minutes (baseline), test compounds were added to the perfusion lines for a period of about 15 minutes. The perfusion was then changed back to regular ACSF.

Paired-pulse stimulation was used because stimulation of the SC fibers, in part, activates interneurons which generate an inhibitory postsvnaptic potential (IPSP) in the pyramidal cells of CAl. This feed forward IPSP typically sets in after the EPSP reaches its peak.

It accelerates the repolarization and shortens the decay phase of the EPSP. and thus could RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. 11 2 613 9663 3099 38 partially mask the effects of the test compounds. One of the relevant features of the feed-forward IPSP is that it can not be reactivated for several hundred milliseconds following a stimulation pulse. This phenomenon is employed to advantage to eliminate IPSP by delivering paired pulses separated by 200 milliseconds and using the second ("primed") response for data analysis.

The field EPSP recorded in field CAI after stimulation of CA3 axons is known to be mediated by AMPA receptors: the receptors are present in the synapses [Kessler et al..

Brain Res. 560: 337-341 (1991)] and drugs that selectively block the receptor selectively block the field EPSP [Muller et al.. Science. supra]. Aniracetam increases the mean open time of the AMPA receptor channel and. as expected from this. increases the amplitude of the synaptic current and prolongs its duration [Tang et al. Science. supra]. These effects are mirrored in the field EPSP. as reported in the literature [see. for example. Staubli et al.. Psvchobiolog'. supra: Xiao et al.. Hippocampus supra; Staubli et al.. Hippocampus 2: 49-58 (1992)]. Similar results have been reported for the previously disclosed stable benzamide derivatives of aniracetam [International Patent Application Publication No. WO 94/02475 (PCT/US93/06916) (Lynch and Rogers. Regents of the University of California)].

The characteristic of a compound to produce an increase in the EPSP response has been a reliable predictor of the ability to improve memory in the 8-arm radial maze task.

Furthermore, a reliable increase in the amplitude, but not the half-width. of the EPSP response is the hallmark of a compound that is efficacious in animal models of schizophrenia. As a nonlimiting example. the ECo values for the actions of CX516 and CX691 to increase the amplitude of the field EPSP are 180 .M and 3 pM. respectively. The increased potency in the in virro slice model is mirrored in the comparative efficacies to reverse the effects of methamphetamine in an animal model of schizophrenia, as discussed below.

RECEIVED TIME 28. JUN, 11:07 PRINT TIME 28. JUN. 11:24 -613 9663 3099 39 Example 2 Svnerev Between an All1o-steric Potentiator of AMPA Receptors and Clozapine in an Anima Model of Schizophrei Amphetamine induction ot'stereoty'pic behavior is a well-known and widely used animal imodel of schizophrenia. The logic for this has been based primarily on two related sets of findings: 1) Amphetamine abuse in humans is known to provoke psychotic symptoms inicluding paranoid ideation. delusions. hallucinations, and stereoviypcd compulsive behaviors: and.

2)Antipsychotic drugs that are effective in the treatment of human schizophrenia 3re also known to attenuate stereotypic behaviors induced in rats by amphetamines.

Finding no. indicates that amphetamine- induced stereotypic behaviors in rats arc a useful mode] for screening potential anti-schizophrenic drugs. Both findings have been instrumental in validating the hypothesis that psychotic symptoms are due, in part. to hyperactive dopamninerttic transmission since amphetamines enhance dopamnine release and typical neuroleptic drugs arcpotent dopamine receptor antagonists. The experiments described below Used enhanced locomotor and stereotypic rearing activity induced by amphetamines in rats as a model. Published authoritv for the use and reliability of this model is found in; .Janssen.

Li at.. Is it possible to predict the clinical effects of neuroleptic drugs (major tranquilizers) from animal data'? TV. An improved experimental desisin for measuring! the inhibiton- effects of neuroleptic drugzs on armphetamine- or apomnorphine-induced "Cheroing& and "agitation' in rats" .Arz:;7imiIl'CI-Forx.chuni~g 17:841-854 (1967); Bentall. A.C.C. er al., "Blockade of amphetamineinduced locomotor activity and stereotypy in rats by spiroperidol but not by an atypical thioridazine.' A-europharmacolop' 19:699-703 (1980): Niemnegeers. ei al..

"A systematic study of the pharmacological activities of dopamine antagonists," Life Science 24:2201 -22]16 (1979): and Homvki6ewicz. -Psychopharmacological implications of dopaminie and dopamnine antagonists: a critical evaluation of current evidence." A.etroscience 3:773-783 (1978).

RECEIVED TIME 28. UN. 11:07PRN TIE 2.J .114 PRINT TIME 28. JUN. 11 2 4 613 9663 3099 In the present experiments, male Sprague-Dawley rats (250-300 g; Charles River Laboratories) were given ad libitum food and water and maintained on a 12:12 hr light:dark cycle with lights on at 6:00 AM. Behavioral studies utilized a computerized Photobeam Activity System (San Diego Instruments. San Diego. CA). in which each of ten test cages (standard S polycarbonate animal cage; 26 cm x 48 cm x 20 cm: W x L x H) were surrounded by two photobeam arrays that were placed to detect locomotor behavior with a lower array and rearing behavior with an upper array. Locomotor and rearing activities were continuously monitored by computer for all ten test cages. Test cages (with photobeam arrays) were placed in a partially darkened room with room ventilation as background noise. On the test day, naive rats were initially placed in the test cages and baseline behavioral activity in the novel environment was monitored during a 30-minute acclimation period. The rats were then injected with vehicle or dru(s) dissolved in vehicle and immediately returned to the test cage and monitored undisturbed for 90 minutes.

Experimental groups (n 10-12. except vehicle, n 6) were 1) vehicle (saline or 1% lactic acid. pH 2) S-(+)-methamphetamine HCI (METH: 2.0 mg/kg); 3) METH (2 mg/kg) CX516 (10 mg/kg); 4) METH (2 mg/kg) clozapine (1.0 mg/kg); and 5) METH (2 mg/kg) CX516 (10 mg/kg) clozapine (1.0 mg/kg). Behavioral experiments were repeated at least twice for each condition. Photobeam breaks were summed by the computer into ten-minute periods for analysis. Group means and standard errors are reported in the figures: means and standard deviations were used for statistical analysis by unpaired, two-tailed t test assuming unequal variance.

Activity measurements presented in Figures 1 and 2 show that CX516 synergistically enhanced the antagonistic activity of clozapine (1.0 mg/kg). a commonly-used atypical antipsychotic. in the methamphetamine animal model described above. Clozapine mg/kg) alone had no effect on METH-induced rearing activity, whereas CX516 mg/kg' caused a modest but statistically insignificant, antagonism of METH-induced stereoiypic rearing. However, together, the combination of clozapine and CX516 acted synergistically and greatly reduced METH-induced rearing activity during the 90-minute test period. After correction for the rearing activity of vehicle-treated control rats, the clozapine/CX516 combination reduced METH rearing activity by RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. I I 2 613 9663 3099 41 In an additional experiment, the combination of CX5 16 (10 mg/kg) and another atypical antipsychotic. risperidone (0.1 mgakg), appeared Lo be synergistic by completely reducing METH-induced rearing to the vehicle level (100% reduction): whereas each agent S alone reduced rcaringy by 28% and 51%. respectively. (p 0.01 vs METH- RiSP)(see Table 1 for a tabular compilation of representative synergistic interactions between Ampakines and antipsychotics).

Example 3 Svnerav between an Ampakine and Halopeiol in the Metharnphetarnine Hyveractivity Animal Model of Schizop~hrenia Using the same methods described in the previous example. the test drug was with the commonly-used typical neuroleptic. halopericlol. As shown in Figs. 3 and 4, haloperidol (0.06 mg/kg) or CX5 16 (30 mg/kg) each produced modest, non-significant antagonism of METH-induced stereotypic rearing activity of 13% and 22%. respectively.

However. the combination of the same doses of haloperidol and CX5 16 was synergistic.

reducing METH--induced rearing activity more completely than the sum of the effects of either druta alone: 67% vs 37%. Analysis of the difference between the effect of the haloperidol/CX5I6 combination and haloperidol alone by two-tailed, unpaired tctst was highly signifficant (p 0.0005). This same dose combination also produced a synergistic effect when locomnotor activity (LMA) was measured: 9% or 10% reduction of METH- LMA for haloperidol (0.06 mg/kg) or 16 (30 mg/kg), rcspectively, whereas the drug combination reduced METH- LMA by 56% 2$(p 0.00 5 vs METH HAL 0.06 mg/kg).

Table 1. Percent Rcduction in Methamphetamnine Induced Activitv Antipsychotic (dose Activity Antipsychotic 1CX516 alone CXSI16 plus in mg/Jkg) alone J_______Antipsychotic Haloperidol (0.06)1 Rearing 16 23 71 Crossing 9 10 Haloperidol 12) Rearing 69 43 84 Crossing 1 66 43 1 76 RECEIVED TIME 28- JUN. 11:07 PITTM 8JN 12 PRINT TIME 28AUN, 11:24 613 9663 3099 42 Fluphenazine Rearing 57 37 81 LCrossing 54 47 79 Clozapine IRearing -5 34 I_ Crossing -6 -32 Risperidone 1) Rearing 51 2810 Crossing 43 0 54 IThe dose of CX516 was 10 mg/kg in all experiments except 30 mgk in0 i ae Example 4 Receptor Interactions The classic typical antipsychotics. such as haloperidol. chlorpromazine fluphicnazine. all have in common the ability to potently antagonize the D2 dopamine receptor and bb, dopamninerigic transmission. Early studies correlated the clinical efficacy of typical antipsychotics v their potency as doparnine receptor antagonists. giving rise to the dopamine hypothesis schizophrenia; Creese er al.. Science 192:481-482, (1976). Newer atypical antipsychotics, suet clozapinc. risperidone and olanzapine generally are potent antagonists at serotonin receptors, but r still be relatively potent antagonists at dopamnine receptors.

On the other hand. Ampakines, typified here by CX5 16 and CX69 I. are quite spec, for ANIPA-type glutamate receptors. Table 2 presents the results of radioligand binding studies 1 show a lack of interaction between Ampakines and dopamninergic or serotonergic receptors. Thus.

skilled in the art would not expect that either additive or synergistic effects would result upon administration of an Ampakine with a typical or atypical antipsychotic.

RECEIVED TIME 28A UN. 11:07PRN TIE 2.J.1:4 PRINT TIME 28-JUN, 11:24 -613) 9663 3099 43 Table 2. Radioligand Binding Analysis of Potential Interactions between Select Ampakines and Select Neurotransmiitter Receptors N eurotransmitter Adrenergic Dopaminergic Muscarinic Serotoninerg-ic Adreneraic D op amin e Muscarinic Seroto in Radioligand (3H-f-Prazosin [i-lJ-Spiperone [5H]-QNB [-'H]-dLSD [-'H]-Prazosin [31H]-Spiperone 3

H]-QNB

3 H]-dLSD Ampakine CX516 CX5 16 CX516 CX5]6 CX691 CX691 CX691 CX691 Concentration (Miolar) I E-5 I E-7 I E-9 I E-5 I E-7 I E-9 I E-5 I E-7 I E-9 IE-5 i E-7 I E-9 I E-4 I E-7 1 E-4 I E-7 I F-4 I E-7 I E-4 I E-7 Inhibition of ligand binding -0.2? 0.0 -2.8 0.6 -3.1 0.4 9.1 9.9 .0 10.7 6.1 -3.7 2 1.6 Example Administration to humans A first step in treating humans is generally determining that a particular patient exhibits the symptoms of a psychotic behaviour such as Schizophrenia or S chizoplireni formn Disorder or Schizoaffective Disorder or Delusional Disorder or B~rief Psychotic Disorder or Psychotic Disorder Due to a General Medical Condition or Psychotic Disorder Not Otherwise Specified. This determination is made by a person skilled in the art using a number of readily available diagnostic proccdures. In general. the presence of Typical DSMJV psychotic dysfuinctions in humans can be ascertained via observation. diagnosis.' family history.

questionnaires or intervicws. The success of treatment is measured by monitoring and recording the abatement of the symptoms of the treated behavioral disorder.

RECEIVED TIME 28. UN. 11:07PRN TIE 2.J 1:3 PRINT TIME 28AUN. 11:23 613 9663 3099 44 In addition. the present invention provides for kits with unit doses of AMPA upmodulating drugs and neurolepiics either in oral or injectable doses. In addition to the coniainers containin2 the unit doses will be a Informational package insert describing the use and attendant beniefits of the drues in treating neurodegenerative pathologics not Saffectins-i memory or learning. Preferred compounds and unit doses include those described herein above.

RECEIVED TIME 28. UN. 11:07PRN TIE 2.J. 1:3 PRINT TIME 28. JUN. I I 2 3

Claims (3)

1-i. OH, and C I-C 6 alkoxy; 21 P, is either combined with R 4 or is a member selected from the group consisting of 22 H. 01H. CI-C 6 alkoxy. amino, mono(C 1 -C 6 alkyl)arnino. di(Cf-C 6 alkvl)amino. 23 and CH 2 OR9. in which R 9 is a member selected from the group consisting of 24 H. CI-C 6 alkyl. an aromatic carbocyclic moiety. an aromatic heterocyclic moiety, an aromatic carbocyclic alkyl moiety, an aromatic heterocyvclic alkyl 26 moiety- and any such moiety substituted with one or more members selected 27 from the group consisting of C i-C 3 alkyl, CI-C-, alkoxvy. hydroxy, halo, amino. 28 alkylaxnino. dialkylamino, and methylenedioxy; 29 R 6 is either iI or CH- 2 OR 9 R and R4" when combined form a member selected from the group consisting of 31~ N(CR 2 P 2 2q_1) N 32 N 8 C \R q Zq-1~ and R~1 33 in which-, 34 R' 0 is a member selected from the group consisting of 0, NH and N(C 1 -C 6 .alkyl); 36 R1 1 is a member selected from the group consisting of 0, NH and N(C I-C 6 37 alkyl); 38 R 1 2 is a member selected from the group consisting of H- and C alkyl, 39 and when two or more R 1 2 ,s are present in a single compound, such R1 2 s are the same or different: 41 pis1. 2, or 3.and 42) q is Ior 2:and RECEIVED TIME 28. JUN, 1hD RITTIE2.07. 12 PRINT TIME 28-JUN. 11:23 613 9663 3099 47 43 R 7 when not combined with any R 8 is a member selected from the group consisting 44 of H. CI-C 6 alkyl, and CI-C 6 alkoxy. 1 6. A method in accordance with claim 5 in which R 2 is a member selected from the 2 group consisting of (CHR),.m and Cn.mHRS2,n-m)J, and R j is a member selected from the 3 group consisting of H. Ci-C 6 alkyl, and Ci-C 6 alkoxy. 1 7. A method in accordance with claim 5 in which R 2 is a member selected from the 2 group consisting of (CHR)n-m and Cn.mHR 2(n-m-3, and one R 8 is combined with either R 3 or 3 R 7 to form a single bond linking the no. 3' ring vertex to either the no. 2 or the no. 6 ring 4 vertices or a single divalent linking moiety linking the no. 3' ring vertex to either the no. 2 or the no. 6 ring vertices, the linking moiety being a member selected from the group consisting 6 of CH,. CH 2 -CH 2 CH=CH, O, NH. N(Ci-C 6 alkyl),N=CH. N=C(C -Cs alkyl). O-C(O), 7 CH(OH), NH-C(O), and N(C,-C 6 alkyl)-C(O). 1 8. A method in accordance with claim 5 in which R 2 is a member selected from the S group consisting of (CHR')nm and Cn-mHR2(n-m)-3, and one RR is combined with either R 3 or 3 R 7 to form a single bond linking the no. 3' ring vertex to either the no. 2 or the no. 6 ring 4 vertices or a single divalent linking moiety linking the no. 3' ring vertex to either the no. 2 or the no. 6 ring vertices. the linking moiety being a member selected from the group consisting 6 of CH2. CI-I-CH_, CH-CH, O. NH. and CH(OH). 1 9 A method in accordance with claim 5 in which R 2 is a member selected from the 2 group consisting of (CHR 8 and Cn-mHR 8 2(n-m)3, and one R 8 is combined with either R 3 or 3 R 7 to form a single divalent linking moiety linking the no. 3' ring vertex to either the no. 2 or 4 the no. 6 ring vertices, the linking moiety being a member selected from the group consisting of CH2. O. NH. and CH(OH). 1 10. A method in accordance with claim 5 in which m is zero. R 2 is a member selected 2 from the group consisting of CHR-CH2-CH2-CH 2 and CHR 8 -CH 2 -CH 2 -CH 2 -CHz, in which 3 R 8 is combined with R 7 to form a single divalent linking moiety linking the 2 and 3' ring 4 vertices, the linking moiety being a member selected from the group consisting of CH-. O, NH. and CH(OH). 1 11. A method in accordance with claim 5 in which m is zero. R 2 is a member selected 2 from the group consisting of CHR 8 -CH 2 -CH 2 -CH2 and CHR'-CH 2 -CH 2 -CH 2 -CH 2 in which 3 R 8 is combined with R 7 to form a single divalent linking moiety linking the 2 and 3' ring 4 vertices, the linking moiety being a member selected from the group consisting of CH. O. and NH. RECEIVED TIME 28. JUN, 11:07 PRINT TIME 28. JUN. 11:23 613 9663 3099 48 1 12. A method in accordance with climir 5 in which R4 and R" are combined to form a 2) member selected from the group consisting of R 1 1 R 3 C 2 PC q R22q-1 N RO R 12 N 4q 2q- I and R 1

13. A method in accordance with claim 12 in which is a member selected from 2 the croup consisting of H- and CH 3 1 14. A method in accordance wvith claim 5 in which R and R. are combined to form a 2 member selected from the group consisting of (CR' 2 Y an 112 C 4 in which R' 2 is a member selected from the group consisting~ of H and Cl- 3 1 15. A method in accordance with claim 5 in which: 2 mn is zero-, 3 R 2 is a member selected from the group consisting of.Cl1RK-CH- 2 -CH- 2 -CH4 2 and 4 CHR'-CH1 2 -CH,-CH,-CH 2 in which R" is combined with R' to form a single divalent linking moiety linking the 2 and 3 ring vertices. the linking moiety 6 being a member selected from the group consisting of CH 2 0, N1H. and 7 CH(OH): 8 R-1 and R i are combined to form a member selected from the group consisting of R 1 1 1- 12( K- 12 9 (CR 2P q R2q-1 RECEIVED TIME 28. UN. 11:07PRN TIE 2.J. 1:3 PRINT TIME 28. JUN. I I 2 3 613 9663 3099 49 N N N 12 (C R 12 11 2q-1 and R 1 and 11 R' 0 is O; 12 R" is O0 13 R 1 2 is a member selected from the group consisting of H and CH 3 1 16. A method in accordance with claim 5 in which R 4 and R' are combined to form a 2 member selected from the group consisting of N R 11 N C C R 10 1 3 (CR 2 2 P and 1 4 in which R' n is O, R" is 0, and R is a member selected from the group consisting of H and CH 3 1 17. A method in accordance with claim 5 in which: 2 m is zero; 3 R 2 is a member selected from the group consisting of CHR'-CH 2 -CH2-CH2 and 4 CHR'-CH2-CH_-CH,-CH2, in which R 8 is combined with R 7 to form a single divalent linking moiety linking the 2 and 3' ring vertices. the linking moiety 6 being a member selected from the group consisting of CH 2 0, and NH; 7 R 4 and R 5 are combined to form a member selected from the group consisting of N S(R12 P and 12 and 9 RO is O; R' is O; and 11 R 1 2 is a member selected from the group consisting of H and CH;. 1 18. A method in accordance with claim 5 in which: 2 m is zero; 3 R' is N; RECEIVED TIME 28. JUN. 11:07 PRINT TIME 28. JUN. I I 2 613 9663, 3099 4 R 2 is CH1--CI- 2 -CI1 2 -CH 2 R 3 is H; 6 R3 is H; 7 R' and R 5 are combined to formn 1112 8 1 9 R is H; R 8 is combined with R 7 to form a single 0 atom linking the 2 and Y ring vertices; I] and 12 R' 2 isH. 1 19. A method in accordance with claim 1, wherein the first compound has the 2) following structure: H NSO 2 SA 2 1N R 2 4 122 4 wherein R 2 is either H. halo or CE 3 I 6 R~and e~ either are both H or are combined to form a double bond bridging the 3 7 and 4 ring vertices; 8 R 24 is either H. C 1 -C 6 alkyl, C 5 cycloalkyl, C,--C7 cycloalkenyl. Ph. CH 2 Ph, 9 CH2SCH 2 Ph, C-HX, CHX 2 CH 2 SCH-2,CF 3 CHiSCH 2 CH-CHi. or l 11 and Re is a member selected from the group consisting of H and Cj-C 6 alkyl. RECEIVED TIME 28. UN. 11:07PRN TIE 2.JN 1:3 PRINT TIME 28AUN, 11:23 613, 9663 3099 1 20. A method in accordance with claim 1, wherein the first compound has the 2 following strucitre: R R 4 in which: 6 R1 is oxygen or sulfur; 7 R 2 and R are independently selected from the group consistIng of and 8 CX=, 9 M is or mCR' wherein R 4 and R8 are independently R or together form a single linking moiety linking M to the ring vertex the linigoetbigslcedrmth 11 group consisting of a single bond. -CR 2 -CR CR-, -S(COX-. and 12 R5 and R 7 are independently selected from the gyroup, consisting of -(C 2 13 CR=CR-. -CR=CX-. -CX 2 and and 14 R is selected from the group consisting Of -(CR 2 -CRCR., and 16 wherein 17 X is -Br. -Cl. -CN. -NO 2 -OR. -SR. -CO 2 R. or -CONR 2 18 and 19 R is hydrogen. C,-C 6 branched or unbranched alkyl, which may be unsubstituted or substituted with one or more furictionalities defined above as X. or aryl. which may be 21 unsubstituted or substituted with one or more fictionalities defined above as X 22? m and p are independently 0 or 1, 23 11 and y are independently 0, 1 or 2. 1 21. The method of claim I wherein the second antipsychotic compound is selected 2 from the group consisting of typical and atypical antipsychotic compounds. 1 22. The method of claim 21 wherein the typical antipsychotic compound is selected 2 from the group consisting of haloperidol. chlorpromazine. fluphenazine. perphenazine. 3 molindone. pimozide. trifluoperazine and thioiridazine. RECEIVED TIME 28. UN. 11:07PRN TIE 2.J. 1:3 PRINT TIME 28. JUN. I I 2 3 613 9663 3099 52 1 23. The method of claim 21 wherein the atypical antipsychotic compound is selected 2 from the group consisting of clozapine. risperidone, olanzapine, sertindole. M100907. 3 ziprisidone. seroquel. zotepine, arnisuipride. and iloperidone. 1 ~24. The method of claim 1 wherein the second antipsychotic compound is 2 administered at subtherapeutic levels. 3 25. A method in accordance with claim 5, wherein the first compound has the 4 following structure: 0 N

26. A method in accordance with claim 20 in which the first compound is selected 2from the group consisting of the following compounds: 0 N N 0 N 0N Na OH 0 N 0 NN N 3 CN RECEIVED TIME 28. JUN,. 1: 07 PRINT TIME 28. JUN. I11:23