CA2195239A1 - Optically active isomers of dihydrexidine and its substituted analogs - Google Patents

Abstract

Optically active hexahydrobenzo¢a!phenanthridines of formula (I) wherein R is hydrogen or C1-C4 alkyl; R1 is hydrogen or a phenol protecting group, X is fluoro, chloro, bromo, iodo or a group of the formula OR5, and R2, R3, and R4 are independently selected from the group consisting of hydrogen, C1-C4alkyl, phenyl, fluoro, chloro, bromo, iodo, or a group -OR1 are disclosed. The method of resolving the racemic trans-hexahydrobenzo¢a!phenanthridines into their component enantiomers is also disclosed. Pharmacological evidence reveals that only one of the enantiomers of a preferred phenanthridine, dihydrexidine, the (6aR, 12bS)-(+)-isomer, is active in binding to both D1-like and D2-like dopamine receptors. Various other compounds, compositions, and methods of using the optically active stereoisomers are likewise disclosed.

Description

~ 096ro2sl3 2 ~ 9 5 2 3q OPTICALLY ACTIVE ISOMERS OF
I~I HYIIII~ N~: AND ITS 6U~;L1LIJL~ ANAIOGS

G~v~ t Fundi g Aspects of the present invention were supported by PHS Grants MH427Q5, MH40537, Center Grants HD03310, M~33127, and Training Grant GM07040. The United States Guv t has certain rights in the present invention.

1. Field of the Invention The present invention relates to optically active 8 t e r e o i s o m e r s o f ~t r a n 3 -dihydroxyhexah~d~ubu-lzo[a]r~n~nthridine (dihydrexidine), their resolution, compo3itions, and methods of use. In particular, the (+)- and (-)-isomers of dihydrexidine were resolved from the racemate using a non-classical technique after initial attempts using classical techni~ues failed. Furthermore, pharmacological evidence suggests that the biological activity previously associated with the racemate, with respect to both Dl-like and D2-like ~npom;n~ receptors, resides in only one of the stereoi30mer3, the (+)-isomer. Because each of these receptors constitutes several di~ferent molecular forms, each of which in turn is encoded by distinct genes and is associated with opposite h;nnhPm;cal effects, i.e., D1-like for stimulation vs. D2-like for inhibition oi cAMP synthesis, the discovery that just one o~ the stereoisomers is active in both types of receptors is indeed surprising. The present invention finds use in a number of diagno3tic, prophylactic, and therapeutic applications, ;nnlll~;n~ the alleviation or modulation of certain neurological disorders.

Wo96~2~13 2 ~~

2. Background of the Invention Dopamine functions as a n~uL~Ll~--smitter in both the~
central and peripheral nervous systems. ~~ ~or~m; n~
receptors have been implicated in several neurological disorders, such as schizophrenia and F~rkinRnn's disease, as well as in vascular regulation~ Additionally, ~n~minP receptors are the accepted loci of; action of many psychotropic drugs, including amphetamine, cocalne, and the neuroleptics. Thus, ligands selective for dopamine receptor9 are important as basic research tools and potential therapeutic agents.
Although the ~ield i~ in a state of flux, the~
commonly accepted classification divides dopamine receptors into two general classes called Dl and D2 (,7~h~h;~n, J. and Calne, D.B., in Nature 1979, 277, 93-96), with each group comprising several molecular forms.
The "Dl-like" receptors include at least two gene products (the Dl~ and the Dl~ or Ds)~ which are linked functionally to stimulation of c~MP synthesis, and which preferentially recognize l-phenyl-tetrahydrnh~n7~7~rines ~e.g., SC~21390) vs. benzamides (e.g., raciopride or sulpiride). The "D~-like" receptors come from at least three genes, and include multiple splice variants. The IlD~-like~ receptor9 (D,l0~2, D2~ort~ D~ and D~) 5l -~r--5 are linked to inhibition of c~MP 9ynthesis, and have the opposite pharmacological specificity from the D~-like receptors (i.e., having much higher af~inity for, ~piperone or sulpiride vs. SCH2339C).
Because of a prevailing view that D,-like receptors were responsible for almost all of the rlin;n~lly important actions of ~nr~min~ agonists and antagoni6ts, Dl-like receptors received little attention until the mid-1930s. With the availability of SCH23390 (the first Dl selective antagonisti see~ lorio, ~.C. et al., in J.
ph~rr-cnl. Exp. ~her. 1983, 2?6, 462-468), it soon became, clear that the Dl-receptor had profound psychopharmaco-j -~o 96/02~13 2 1 9 ~ ~ 39 r~ 5 logical effectc and in~r~ F~l in important ways with D, receptors. (Mailman, R.B. et al., in E~ur. IJ. ph~ ro7, 1984, 101, 159-160; Christensen, A.V. et al. Life Science 1984, 34, 1529-1540.) Yet while several ~ Pll~nt antagonists were made from the 1-phenyl-tetrahydro-3-benzazepine series, the resulting agonists (e.g., SKF38393, whose structure is depicted below) were generally only of partial efficacy relative to ~lnp~m;n~
(i.e., in stimulating ~ p~m;n,o-sensitive cAMP synthesis) .

' . ~

1~0 ~

Despite this potential limitation, SKF38393 and related partial agoni~ts have been used in many studies of Dl receptor function because they were relatively selective, and because no alternatives were available.
Nichols, D. E. disclosed, in U.S. Patent No.
5,047,536, novel ligands for rl~lp~m;nP receptors which comprised generically certain trans-hexahy-l, ub~ oph~n~nthridines of the formula (1) 1~5 7 b wherein Ho and Xb are trans across ring fusion bond c, R is hydrogen or C1-C~ alkyl; Rl is hydrogen, benzoyl or W096/02513 ' ~ ~.9s ? 3 9 ~ S~

pivaloyl; and ~ is hydrogen, chloro, bromo, iodo or a~
group of the formula OR, wherein Rl i5 hydrogen, benzoyl or pivaloyl.
The biological activities of these compounds were S described as ranging from potent dopamine-like activity affecting both D-1 and D-2 dopamine reçeptor subtypes to specific dopamine D-1 receptor ~nt~gnn; Rt activity.
Although numerous compounds exemplifying the generic class are provided, the speci~ication of this patent ~nnt~;nq neither a description of the stereoisomers of any of the compounds nor of their potential pharmacologic behavior.
These previous studies cnlm;n~ed i~ the synthesis of racemic trans-10,11-dihydroxyhexahydrobenzo[a]phen-anthridine (dihydrexidine, 2), the first compound shown to be a potent and bioavailable, full efficacy D
agonist. (Brewster, W.K. et al., in J. ~ed. Chem. 1990, 33, 1756-1764; ~ovenberg, T.W. et al., in Eur. J.
ph~rr~col . 1989, 166, 111-113.) In Brewster, W.K., supra, the authors speculated on what possible activities may reside in one enantiomer of dihydrexidine versus another. Because no studies had actually been carried out on the individual enantiomers, such speculation only highlights the uncertainty of what properties each ~n~n~ r may or may not exhibit.

~o ,1~ ~ J
In addition to its use as a pharmacological probe, 2 has been shown to have impressive antipark;n~nn;~n action in the MPTP monkey model. (Taylor, J.R. et al., in Eur. J. Pharmacol. 1991, 199, 389-391.) ~WO96/02513 2 1 9523~

More recently~ similar a~tiparkinsonian effects were reported for members of another new class of full D
agonists, the phenyl aminomethylisochromans. (DeNinno, M.P. et al., in ~. Med. Chem. 1990, 33, 2948-2950.) The structure of one such compound, (lR,35) -1- (~m;n~ thyl)-3,4-dihydro-5,6-dihydroxy-3-phenyl-lH-2-benzopyran (A70108) is shown below.
[~

Such data underscore the importance:of understanding the pharmacophore (thoae interrelated steric and electronic features that impart a specific pharmacological character to a particular drug) for the Dl-like receptors.
All previous studies involving the ~ _-ulld 2, both in vivo and i~ vitro, have used the racemate. Moreover, while (~)-2 is selective for ~np~inP receptors, it has only about ten-fold selectivity for Dl vs. Dl receptors in rat striatum. (See, Brewster, W.K. et al., supra;
Mottola, D.M. et al., in ~. ph~rr-co7. ~xp. Ther. 1992, 262, 383-393.) Of particular interest is the unprece~Pnted selectivity of 2 for activating functions mediated by postsynaptic, but not presynaptic, D2-like receptors. (See, Mottola, D.M. et al., supra; also, Mottola, D.M. et al., in Soc. Neurosci. Abstr. 1991, 17, 818 and Nichols, N.F. et al., in Soc. Neurosci. Abstr.
1992, 18, 1170.) Mottola and co-workers (1992?, supra, based on molecular I ~Pl ing studies, hypothP~izP~ that a particular Pn~nt;, of dihydrexidine, (6a~,12bS), would W09~0~13 2 ~ 9 S 23 ~ 6 P~

be the more active Dl ligand. Again, no work was~
actually carried out using individual enantiomer3, and the authors' hypothesis amounts to no more than speculation. MJL~UV~L, in which ~n~nti,: r the D, activity might lie was, until the present work, very much anyone's guess.
Thus, knowledge of which rn~nt;~ r or enantiomers iB responsible for the D1 and D~ activities of 2 is of critical importance in understanding the intimate aspects;
of the D~ and D2 pharmacophore and of ne~rological processes ~ tPd by dopamine receptors, in general.
Certain substituted rh~n~nthridines are disclosed in International Publication Number W0 92/04356. At page 11, of the disclosure, the specification states the obvious in that a ~ ~ ~ having two or more asymmetric carbon atoms exists in diastereomeric forms. Stating that the alleged invention included within its ~cope all of the isomeric forms of a claimed compound, the specification once~against recites a generally accepted proposition. Hc~ever, on closer ~m;n~tjon, the specification does not contain specific disclosures of individual enantiomeric compounds. Moreover, the specification is devoid of any rationale or motivation for performing the resolution of racemic mixtures, let alone include any discussion of a viable means for accomplishing such an operation.

3. 8 = ry of the Invention Accordingly, the present invention provides the resolution of the Pn~nt; r9 of 2 and its substituted hexahydrobenzo[a]ph~n~nthridine analogs and discloses pharmacologic evidence that suggests that, quite unexpectedly, both D1 and D, receptor affinities, as well as functional effects, ;nrln~1nr; the unprrr~dent~ D, postsynaptic selectivity, reside in the (6aR,12bS)-(+)-.

~WO96/0~13 2 1 ~239 }~

enantiomer of 2 and the corr~ap~n~in~ ~nantil ~ of its substituted analogs.
Generally, then, it is an object of the present invention to provide a compound of the formula R ~ R4 I
R~O ~ ~ R

and pharmaceutically acceptable salts thereof wherein the X. and X~ are trans across ring fusion bond c, R is hydrogen or C1 - C, alkyl R1 iS hYdL~Y~1~ or a phenol protecting group X i8 fluoro, chloro, bromo, iodo, or a group of the formula -ORs wherein Rs i9 hydrogen or a phenol protecting group, provided that when X is a group of the formula -ORs, the groups R1 and Rs can be taken together to form a group of the formula -CX,-; and R " R3 and R, are independently selected from the group consisting of hydrogen, C1-C~ alkyl, phenyl, fluoro, chloro, bromo, iodo, or a group -OR1 wherein R1 is as defined above, provided said ~ ~ ' is optically active. Preferably, the compound is the (+)-isomer, although in other --; a, the compound is the (-)-isomer. In a particular: '10~i t of the invention, the groups R2, R " and R~ are all hydrogen. In another, at least one of the groups R " R3, and R~ is methyl.
Other objects of the present invention, including compositions comprising the disclosed optically acitve compounds and methods of their use, will be apparent to 21 ~239 WO96/02513 .

one of ordinary skill considering the= ~P~;l de3criptions provided herein.

4. srief Description of the Flgures Figure l. Competition of ~n~nti~ ~rs of 2 for D
receptors labeled by 3H-SCH23350. (A) Competition in rat striatal membranes. (B) Competition in ~ dnes prepared from Ltk-cells transfected with human Dl receptor. ~+~-2 had ca. twice the affinity of racemic 2 in both preparations. In the striatal membranes, (-)-2 had significantly less affinity than the racemate or (+) enantiomer.
Figure 2. Stimulation of cAMP synthesis by drugs in rat striatal I ' dnes. (+)-2 was nearly l00 fold more potent than the 5-) enantiomer in stimulating cAMP
synthesis in rat striatal membrane3.
Figure 3. Competition of ~n~nt;t ~ S of 2 for D
receptors in rat striatal membranes labeled by 3H-spiperone. (+)-2 had about twice the affinity of racemic 2, and both the racemate and ~+)-~n~n~i~ ~n had significantly greater affinity than the 1-) enantiomer.
Figure 4. A schematic illustration is provided for the isolation of the enantiomers of dihydrexidine. In the scheme, the lower case letters for each step denote the following reaction conditions: (a) i. (R)-Methoxyphenyl acetyl chloride, 20% NaOH (aq), CH,Cl~;
ii. ch~, tography; (b) LiEt3BH, THF; (c) BBr3, CH2Cl2.

5. Detailcd Description of the Pre~arred r 5.1.RnRolut;rn of the Rnnnt~
Initial attempts to resolve the enantiomers of 2 nt;l;7;ng clas3ical resolution techniques were ineffective. In particular, it was discovered that neither the tartrate nor the dibenzoyltartrate diast~, t c salts of the ether-protected amine precursor 3 (See, Figure 4) could be separated by ~ 096/02513 2 1 9 ~ 2 3 9 cry~t~ll;7ati~n_ ~ The attempts u~ing these classical methods were, therefore, ~h~r~n~ in favor of a different method, described further below.
As depicted in Figure 4, the diastereomeric (R)-0-methyl --n~ acid amides of racemic trans-lo,11-dimethoxy-5,6,6a,7,8,12b-hexah~,d~,,bt:l~o[a]rhPn~nthridine 4 were prepared by an adaptation of the procedure of Johansson, A.M. et al., described in J. Med. Chem. 1987, 30, 602-611. In this procedure, the ether-protected amine 3 was coupled with (R)-0-methyl mandeloyl chloride to yield the two diastel. r; C amides.
The=resulting amides were separated by centrifugal chromatography (chromatotron) using 40~ ethyl acetate/hexane eluent and a 2 mm silica gel rotor. While the chromatron was used to effect this separation on a small scale, any of a variety of ch, trgraphic techniques would apply on larger scales, such as column chromatography. Crystallization of the individual amides provided the diast~ in greater than 99~ purity, as judged by HPBC analysis. An X-ray quality crystal was obtained for the diastereomer with lower Rf on TBC and the analysis demonstrated that it corr~pnnA~d to (6aS,12bR)-4. Reductive cleavage of the chiral ~n~ rieg of (6aR12bS)-4 and (6aS,12bR)-4 with LiEt3BH
(Super Hydride) in THF (see, Mellin, C. et al., in ~.
Me~. Chem. 1991, 34, 497-510) provided the resolved amines (6aR,12bS)-(+)-5 and (6aS,12bR)-(-)-5.
The ~n~nt; -ricallypure ~t~h~l~m;n~ (6aR,12bS)-(+)-2 and (6aS,12bR)-(-)-2 were prepared from the corresponding methoxy precursors by treatment with BBr3 and cryst~ll;7ed as their hydrochloride salts.
It is important to note that in the reductive cleavage step, the use of Super Hydride is critical because, as appIicants have discovered, the chiral auxiliary group is not removed by the usual type of reducing agents, such as lithium aluminum hydride.

WO 96/02513 2 ~ ~ 5 2 3 q 5 . 2 . ph "
The PnAnti rs of 2 were tested for ~np~minPrgic activity using in vitro biochemical techni~ues. The results showed that (+)-2 had about twice the affinity of 1~)-2 for the Dl receptor in both rat striatal ~~ ' ~lles, and in transfected Ltk- cells. In the striatal membranes, the (-)-~n~n ti~ - r shows significantly less affinity than either the racemate or l+)-Pn~nt;~ --Like racemic 2 or ~np~minP~ 11)-2 causes a ~ hl;n~ of lD cAMP synthesis in striatal memhranes, indicating that (+)-2 is a full agonist. Conversely, (-)-2 is nearly 100-fold less potent than ~+)-2, and does not cause a fully maximal response (relative to ~np~m;nP) at the highest concentration tested (5 ~M). In competition for D, receptors, 1+)-2 exhibits about twice the affinity of (+)-2, and about 15-fcld higher affinity than the 1-)-enantiomer. These data indicate that the active enantiomer for both Dl and D2 receptors is 16aR,12bS)-(+)-2.
Turning now to the ~. ;n;ng figures, Figure l illustrates that, in both rat striatum and Ltk--hD
cells, (6aR,12bS)-(+)-2 had approximately twice the affinity (i.e., ICs~ = 5.6 nM) of the racemate (IC6~ =
11.6 nM). The (6aS,12bR)-(-) enantiomer had significantly lower affinity than either (6aR,12bS)-(+)-2 or (~)-2 at the Dl receptor.
The data from the ~np~minp-sensitive adenylate cyclase assay (see, Table 1, below, and Figure 2) demonstrate that the 16aR,12bS)-(+)-~n~n~;, and the racemate are about two orders of magnitude more potent than ~np~m;nP, the latter ~havin~ an ECso of about 5 ~M.
Furthermore, (6aR,12bS)-(+)-2 and the racemate both produce a full agonist response of adenylate cyclase. In contrast, the enantiomer (6aS,12bR)-(-)-2 is significantly less potent (ECs~ = 2.15 mM) in the adenylate cyclase assay and lacks the full efficacy ~ 096/0~13 21 9~2~9 character o~ the (+) antipode. It i8 of note that prototypical Dl agonists such as SKF38393 (BCso = 100 nM) and CY208-243 ~6, pl~ = 6.1, whose structure is depicted, below; see, Seiler, M.P. et al., in ~. Med. Chem. 1991, 34, 303-307J are only partial agonists that cause less than 50~ stimulation of adenylate cyclase.
~_ ' ~ C~3 H

A similar pattern was seen in transfected Ltk-cells. This ~inding is of importance because there are 10no D2-like receptors in these cells. Thus, actions of ~n~nt;( ~ of 2 at Dl receptors do not influence adenylate cyclase studies in these membrane preparations, consistent with our earlier findings. (Cook, L.~. et al., in Soc. Neurosci. Aostr. l991, 17, 606~) ~able l.
Co~parison of Dihydrexidine and ita An-l o~
and Certain Other Dop~~ n~ Agoni~ts at D~ and D, Receptor~ in Rat Striatal ~ ~a3 Adenylate Maximal Dl D2 Cyclase St; l~t;nn Drug Potency~ Potency ECso (nM) of Adenylate ICso (nM) ICso (nM) Cyclase (~ vs. DA) (i)-2 11.6 i 132 i 19 140 nM 95 1 . 0 (+)-2 5.6 i 87.7 i 60 nM 105 1.1 12.2 W096/0~13 Adenylate Maximal D1 D, Cyclase st; l~t;on Drug Potency' Potency ECso (nM) of Adenylate ICso (nM) ICso (nM) Cyclase (~ V8. DA) 2 149 i 11 1,250 i ~1000 nd SC~23390 1.01 i - NA NA
1 .o Domperidone - 2.80 i NA NA
1 . O
NA = not applicable ~these compounds are antagonists).
'All tests were performed as described in the Methods section. Hill coefficients for the agonist binding curves were sign;~ ntly less than 1, and the data thus are expressed as io ICsos eince K1 values~ cannot be determined until the number of binding sites 18 resolved.
To minimize interassay variability, the values ln the tables were from assays in which four or more compounds were run on the same day.
While the data described herein~' Ll~te that the Dl activity resides in the (6aR,12bS)-(+)-~n~ntt~ -r, it was also important to ~t~rm; n~ in which Pn~nti~ the;
significant Dl affinity resided. As shown in Figure 3 ~nd Table 1, here again (6aR,12bS)-(+)-2 had about twice the potency of the racemate, with (6aS,12bR)-(-)-2 having significantly lower affinity (87.7, 132, and 1250 nM, respectively).
Therefore, our data indicate that both receptor recognition and functional efficacy at Dl receptors reside principally in the (6aR,12bS)-(+) stereoisomer.
At D2 receptors, the binding affinity also is c~nt~i in the same isomer. It is anticipated that fnn~t;~n~l efficacy at the Dl receptors resides in the (+)-enantiomer, as well We have indicated previously that the full efficacy properties of dihydrexidine may be due to the presence of the trans ~t~n~d conformation of the ethylamine moiety ~ o ~102~13 2 1 9 5 2 3 ~

or the near coplanarity of the aromatic rings. srewster, W.K. et al., in ~. Med. Chem. l990, 33, 1756-176g. As a consequence of the results of the present work, the enantioselective conceptual model of the D1 receptor appears to be vastly superior to two-dimensional models in accounting for the biological activity of these rl c, R ;n;nr to be addressed, however, is the ramification of the D2-like affinity of dihydrexidine and several of its analogues. Existing models of the phar~rorhrre for various D2-like receptors do not, at least at first glance, easily ~- ' te the pendent phenyl ring ofthe lO,ll-dihyd~u~yb~11zo-[a]rhPn~nth~idine class of ~rp~;nP agonists. Moreover, while dihydrexidine haa affinity for D2-like receptors, recent data indicate that its agonist fnnctional properties appear to be limited to only some subpop~ tion~ of these receptors (i.e., those located post-synaptically). Thus, this drug class may be very useful in understanding ligand interactions with the D2-like receptor family, and may provide important data for incorporation into molecular ~-1; ng studies.

5.3.E'- ~e~t;~l Compo~itions Compri~ing the optically Active C _ '- of the Present Invention As should be apparent, the present invention contemplates compositions comprising the optically active compounds disclosed herein. Preferably, these compositions include pharmaceutical compositions comprising a therapeutically effective amounr of an optically active compound along with a pharmaceutically acceptable carrier.
As used herein, the term "pharmaceutically acceptable" carrier means a non-toxic, inert solid, semi-solid liquid filler, diluent, Pnr~p5111Ating material or fu~ t;~n auxiliary of any type. Some examples of the 2l ~5239 ~ ~
W096/02513 r~

materials that can serve as pharmaceutically acceptable carriers are sugars, such as lactoae, glucose and sucrose, starches auch as corn starch and potato atarch, cellulose and its derivatives such as sodium ~arb~x~ thyl cellulose, ethyl cellulose and cellulose acetate; powdered trArAr~nth; malt, gelatin, talc;
excipients such as cocoa butter and suppository waxea;
oila such as peanut oil, cottonseed oil, sAff1( r_, oil, sesame oil, olive oil, corn oil and aoybean oil; glycola, such as propylene glycol, polyols auch as glycerin, sorbitol, mannitol and polyethylene glycol; estera auch aa ethyl oleate and ethyl laurate, agar; buffering agenta auch as magnesium hydroxide and Al 'nnm hydroxide;
alginic acid; pyrogen-free= water; isotonic saline,, Ringer's solution; ethyl alcohol and phosphate buffer solutions, as well as other non-toxic :compatible bubstancea used in pharmaceutical formulations. Wetting agenta, emulsifiers and lubricants auch as aodium lauryl aulfate and ~-gn~A;~Im stearate, as well as coloring agenta, releaaing agents, coating agent~, sweetening, flavoring and perfuming agents, pre~ervatives and~
antirY;~AntA can also be present in the composition, according to the judgement of the full 1 atrr, ~ 1~A
of pharmaceutically acceptable. Ant;rY;~Ants include water aoluble Ant;rY;~Antq auch aa aacorbic acid, cySteine hydrochloride, sodium b;A~llfite, sodium metabisulfite, sodium sulfite, and the like; oil soluble antioxidants auch as ascorbyl palmitate, butylated hydroxyanisole (B~A), butylated hydroxytoluene (BHT), lecithin, propyl gallate, aloha-tocopherol and the like:
and the metal rh~l At; ng agents such as citric acid, ethyl~n~ m;nP tetraacetic acid ~EDTA), sorbitol, tartaric acid, phosphoric acid and the like.
8y a ~'therapeutically effective amount" of an optically active ~ _ JUlld, such as a dopaminergic agent, is meant a sufficient amount of the ~ ~ ' to treat ~Wo9610~13 21 9523~

neurological or behavior disorders at a rP~rnn~hlP
benefit/risk ratio applicable to any medical treatment.
It will bP lln~Prstood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical i'l~. . The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors ;nrl~;nJ the disorder being treated and the severity of o the disorder; activity of the speciiic compound employed;
the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidently with the specific compound employed; and like factors well known in the medical arts.
The total daily dose of the optically active compounds of the present invention administered to a subject in single or in divided doses can be in amounts, for example, from 0.01 to 25 mg/kg body weight or more usually from 0.1 to 15 mg/kg body weight. Single dose compositions may contain such amounts or submultiples thereof to make up the daily dose. In general, treatment regimens ~rrnr~; nJ to the present invention comprise administration to a human or other mammal in need of such treatment from about 1 mg to about 1000 mg of the compound(s) of this invention per day in multiple doses or in a single dose of from 1 mg, 5 mg, 10 mg, 100 mg, 500 mg or 1000 mg.
The compounds of the present invention may be administered alone or in f ' ;n~;on or in concurrent therapy with other agents which affect the central or peripheral nervous system.
~if~uid dosage forms for oral administration may include phar~cent;r~lly acceptable : l~inn~, Wo96~13 21 95239 P~
lÇ
micror~llcinnc~ solutions, suspensions, syrups and elixirs rnnt~in;ng inert diluents commonly used in the art such as water. Such compositions may also comprise adjuvants, such as wetting agents; emulsifying and suspending age~ts; sweetening, flavoring and pel r, ng agents.
Injectable preparations, for example, sterile injectable a~ueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectrable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conv~nt;nn~lly employed as a solvent or snRp~n~ing medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.
The injectable for~ ti~n can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.
In order to prolong the effect of a drug, it is often desirable to slow the absorption of a drug from subrnt~nPnllR or intl Rrnl ~r injection. The moat common way to accomplish this is to inject a suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug becomes ~p~n~nt on the rate of dissolution of the drug which i8, in turn, A~p~n~nt on the physical state of the drug, 21~23q Wo96/0~l3 r~l,u~

for example, the crystal size and the crystalline form.
Another approach to delaying absorption of a is drug to administer the drug as a solution or auspension in oil.
In~ectable depot forms can also be made by forming microcapsule matrices of drugs and biodegradable polymers such as polylactide-polyglycoside. ~Pp~n~ ng on the ratio of drug to polymer and the composition of ~the polymer, the rate of drug release can be controlled.
Examples of other biodegradable polymers include poly-orthoesters and polyanhydrides. The depot injectables can alAo be made by entrapping the drug in iiposomes or microemulsions which are compatible with body tissues.
Suppositories for rectal administration of the drug can be ~ d by mixing the drug with a suitable nonirritating excipient such as cocoa butter and polyethylene glycol which are so1id at ordinary temperature but li~uid at the rectal temperature and will, therefore, melt in the rectum and release the drug.
Solid dosage forms for oral administration may include capsules, tablets, pills, powders, prills and granules. In such 801id dosage forms the active, ~Ju--d may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings and other release-controlling coatings.
Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

Wo96/02513 ~ ~ 9 5 ~3 q The active I uu~ds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be ~uuaL~d with cnat;ngq and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferably, in a certain part of 1~ the intestinal tract, optionally in a delayed manner.
A Of ~ ;ng compositions which can be used include polymeric substanceA~ancd waxes.
Dosage forms for topical or tr~nA~r~ l administration of a compound of this invention further include o;n tA, pastes, creams, lotions, gels, powders, solutions, sprays, ;nh~l~n~A or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophth~lmic formulations, ear drops, eye o; A, powders and solutions are also contemplated as being within the scope of this invention.
The n; ~ ' F, pastes, creams and gels may contain, in addition to an active ~ of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffins; starch, tr~a~~nth~ cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
Powders and sprays can contain, in addition to the compounds of this invention, excipients such as iactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances.
Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.
Transdermal patches have the added advantage of providing controlled delivery of a c _ ~ to the. body.

~ 096102513 2 1 9 5 ~ 3 9 Such dosage forms can be made by dissolving or dispersing the ~ in the proper medium. Abgorption ~nhcn~r5 can also be used to increase the flux of the ~
across the skin_ The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
Accordingly, the present invention is useful in the treatment or alleviation of disease, especially those disorders related to a central nervous system dysfunction. Such dysfunctions of the central nervous system may be characterized by an apparent neurological, phys; nlo~ 1~ psychological, or behavioral disorder, the symptoms of which can be reduced by the administration of an effective amount of the optically active compounds of the present invention.
In particular, the present invention is used in a method of alleviating the effects of P~rkinc~n'5 disease.
A patient can also be treated for a CNS I ~Y~ t-related disorder, including, but not limited to, Huntington's disease, Pick's disease or Creutzfeldt-Jakob disease.
Dihydrexidine has also been shown to be a specific renal vAco~ t~r See, e.g., Kohli, J., in ~urop. ~.
ph~rr-cn7, 1993, 235, 31-35. Xence, the optically active , ~uullds of the present invention can be used in a method of treating or alleviating the effects of a cardiovascular disorder, such as congestive heart failure.
Furthermore, a method of ~nhAn~;ng endocrine function is also r~nt ,1 ~t~ which comprises administering to a patient in need of such ~nh~n~ t an effective amount of the optically active, __ullds of the present invention, especially (6aR,12bS)-(+)-10,11-dihydroxy-5l6l6a~7l~ll2b-hexallyd~ulJellzu[a]rh~n~nthridinel an 0-alkylated or ~-alkylated analog thereof, or its pharmaceutically acceptable acid addition salt, such as WO96/02513 2 ~ 9 ~ 2 3 9 the hydrochloride Preferably, such Pnhnnl leads to an increaee in endocrine function or secretion.
In a specific Pmhc~i-~~ of the invention, a pharmaceutically acceptable antipyschotic composition is provided which comprises an effective amount of the compound (6a~,12bS)-(~)-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydL~b~lzo[a]rhPn~n~hri-dine~ an O-alkylated or N-alkylated analog thereof, or its pharmaceutically acceptable acid addition saIt and a pharmaceutically acceptable carrier.

6. r l9n 6.1.~oltlt;nn of the Rn~nt;. ~ of Dlhydrexidine Using Methods ~ppl;~hle to Its Analog~
6.1.1.Naterials and Methods Melting points were ~Pt~rminPd on a Thomas-Hoover Meltemp apparatus and are uncorrected except where indicated. lH NMR spectra were recorded on a Chemagnetics 200-MHz or a Varian VXR-500S 500-MHZ
instrument. Chemical shifts are reported in values (parts per million, ppm) relative to an intPrn~1 standard of tetramethylsilane in CDCl3, except where noted.
Abbreviations used in NMR analysis are as follows: s, singlet; d, doublet; t, triplet; m, multiplet; dt, doublet of triplets. Analytical thin-layer chromatography (TLC) was performed on Baker-flex silica gel lB2-F plastic plates. Microanalyses were obtained from the Purdue Microanalytical Laboratory and from Galbraith Laboratories, Inc. The chemical inn;7n~i~n mass spectra (CIMS) were de~Prmin~d on a Finnigan 4000 quadrupole spectrometer using ammonia or is~hu~nP as the reagent gas, as noted, and are reported as m/e (relatlve intensity). Optical rotations were obtained on a Perkin-Elmer 241 po1arimeter. Solvents and reagents were used ~WO96102513 2 1 q ~2~q as purcha~ed, except as noted. THF was distilled from nodium metal/b~n7nphpnnnp ketyl.

6.1.2.
(6aR,12bS)-(+)-6-(R-~Y-methG,.y~helylacetyl)-lo~ll-dimethoxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phen-~nt~n;~;n~ )-4, and (6aS,12bR)-(-)-6-(R-a-meth-oxyphenylacetyl)-10,11-dimethoxy-5,6,6a,7,8,12b-hexaL~ L_~zo[a]rh~n-nt~ridine, (-)-4 R- (-) -Methoxyphenylacetic acid ~31 mg, 0.19 mmol;
[~]25-154 degrees; Aldrich Chemical Co.) was added to a 10 mL round bottom flask cnnt~;n;ng thionyl chloride (0.5 mL). After stirring for 2 h at 25 ~C, the vnl~t;l~ were removed. Residual thionyl chloride was removed azeotropically by the codist;ll~t;nn of benzene to provide R- (-) -O-methyl mandeloyl chloride. This residue was dissolved in 1 mL of dichloromethane. Racemic 10,11-dimethoxy-5,6,6a,7,8,12b-hexahydLJb~ u[a]phenan-thridine hydrochloride (50 mg, O.15 mmol) was dissolved in water (1 mL) and dichloromethane (1 mL). After the salt had dissolved, l N NaOH (O. 5 mL) was added, followed by addition of the dichloromethane solution of the mandeloyl chloride.
When TLC analysis (5~ methanol/dichlo,. h~n~, NH
atmosphere) indicated that all of the amine had been Cnn~ 1, the layers were separated, and the aqueous layer was extracted with dichl~rl th~n~ (3 X 5 mL) The pooled dichloromethane fractions were washed se~l~nt;~lly with saturated aqueous sodium carbonate solution (10 mL), 5~ HCl (2 X 5 mL) and brine. After drying over MgSO~, . the dichloromethane solution was filterea and the solvent was removed by rotary evaporation.
The residual oil was then separated into its components using a Chromatotron (Harrison Research, Palo ~ Alto, CA) by elution on a 1 mm silica gel rotor with 40~
ethyl acetate/hexane. The two major fractions were ~nllP~t~ and nnn~ntr~t~d by rotary evaporation. The faster moving c~ _ ~nt was crystA1l;7~d from hexane to W096/0~13 2 1 9 5 2 3 9 provide 16 mg (24%), mp 147-149 ~C; [~D -97.45 degrees (c 0.25, EtOH); CIMS (isobutane): M+1, 444; lXNMR ~ 7.45 (d, 1, ArH, J = 7.6 Hz), 7.38 (m, 2, ArH), 7.33 (m, 3, ArH), 7.21 (m, 1, ArH), 7.00 (s, 1, ArH), 6.97 (t, 1, ArH, J = 7.4 Hz), 6.72 (9, 1, ArH), 6.21 (d; 1, ArCH, J
= 7.41 Hz), 5.15 (s, 1, ArCHO), 4.90 (d, 1, ArCH, J =
14.7 Hz), 4.28 (d, 1, ArCH, J = 14.7 Hz=), 4.20 (d, 1, Ar2CH, J = 12.6 Hz), 3.91 (s, 3, OCH3), 3.83 (s, 3, OCH3), 3.65 (s, 3, OCH3), 3.61 (m, 1, CHN), 3.11 (m, 1, C~Ar), 2.90 (m, 1, CHAr), 2.78 (m, 1, CHCN), 1.66 (m, l, CHCN).
Anal. (C2sH23NO2) Cal'd (Obs'd): C, 75.81 ~75.48); H, 6.59 (6.51~; N, 3.16 (3.16).
The slower running . ~ ~n~ was isolated and crys~ 7ed from ethyl acetate to provide the other diastereomer (20 mg, 30~), mp 171-172 ~C; [~]D +155.91 degrees (c 0.25, EtOH); CIMS (isobutane): M+1, 444; lH
NMR ~ 7.49 (d, 1, ArH, J = 7.69 Hz), 7.44-7.35 (m, 6, ArH), 7.09 (t, 1, ArH), 6.72 (s, 1, ArH), 7.01 (s, 1, ArH), 6.69 (d, 1, ArCH, J = 6.78 Hz), 5.04 (s, 1, ArCHO), 4.86 (d, 1, ArCH, J = 14.4 Hz), 4.17 (d, 2, Ar~CH, ArCH), 3.90 (s, 3, OCH3), 3.83 (s, 3, OCH3~, 3.60 (m, 1, CHN), 3.41 (s, 3, OCH3), 3.11 (m, 1, CHAr), 2.92 (m, 1, CHAr), 2.78 (m, 1, C~CN), 1.66 (m, 1, CHCN). Anal. (C2sH23NO2) Cal'd (Obs'd): C, 75.81 (75.42); H, 6.59 (6.73); N, 3.16 (3.08).

6.1.3.
(6aR,12bS)-(+)-10,11-dimethoxy-5,6,6a,7,8,12b-hexahy-drobonzo[~]rh~nqnf~-idine hydrochloride, (+)-5 The (+)-(6aR,12bS) O-methylmandeloylamide 4 (435 mg, 0.982 mmol) was dissolved in 25 mL of dry THF under nitrogen. The solution was cooled to O ~C and then 6 mL

of a 1.0 M solution of LiEt3BH in THF was added slowly via syringe. The solutiQn was stirred at O ~C for 12 h.
The solution was poured into 25 m~ of ice-cooled 1.O _ HCl. The aqueous layer was washed with 2 x 20 mL ether ~ 096/0~13 2 1 9 5 2 ~ q P~

and then made RlkRl;n~ with NH3. The resulting free base was isolated by extraction with 3 x 30 m~ of dichloromethane. The combined organic extracts were dried (MgSa~), filtered and concentrated in vacuo.
The HCl salt was formed with acidic ethanol and the product was crys~Rll;~ed from Rnetnn;trile to afford 217 mg (67%) mp 232-234 ~C; [~]D +106 degrees (c 0.75, EtOH); CIMS (isobutane): M+1, 296; 1H NMR (d6 DMSO) 9.65 (s, 2, NH2'), 7.44-7.32 (m, 4, ArH), 6.86 (s, 2, ArX), 4.37(d, 2, ArCH,N, J = 4.12 Hz)~, 4.23 (d, 1, Ar2CH, J = 11.07 Hz), 3.75 (s, 3, OCH3), 3.68 (s, 3, OCH3), 3.00-2.94 (ddd, 1, CHN), 2.86-2.79 (m, 2, ArCH2), 2.22-2.15 (m, 1, CHCN), 1.98-1.89 (m, 1, CHCN). Anal.
(C~gH22ClNO2l Cal'd (Obs'd): C, 68.85 (68.79); H, 6.70 (6.71); N, 4.23 (4.24).

6.1.4.
(6~s,12bR)-(-)-10,11-~; ~' y-5~6~6a~7~8~l2b-hexa Lydh~b--zo[n]rhrn~n~hridine hydrochloride, (-)-5 Following an identical procedure to that for (+)-5, the (-)-(6aS,12bR)-O-methyl mandeloylamide 4 (344 mg, 0.777 mmol) gave 173 mg (66%) of the hydrochloride following crystallization from acetonitrile; mp 230-232 ~C; [~]D -107 degrees (c 0.75, EtOH); CIMS (lRnhlltRnP):
M+1, 296; ~H NMR (d6 DMSO) ~ 9.62 (s, 2, NH2~), 7.42-7.32 (m, 4, ArH), 6.86 (s, 2, ArH), 4.37 (d, 2, ArCHlN, ~ =
4.12 Hz), 4.23 (d, 1, Ar2CH, J = 11.08 Hz), 3.75 (s, 3, OCH,), 3.68 (s, 3, OCH3), 3.00-2.93 (dt, 1, CHN), 2.88-2.76 (m, 2, ArCH3), 2.22-2.15 (m, 1, CHCN), 1.98-1.89 (m, 1, CHCN). Anal. (Cl9H~2ClNO2) Cal'd (Obs'd): C, 68.85 (68.82); H, 6.70 (6.66); N, 4.23 (4.18).

6.1.5.
(6aR,12bS)-(+)-10,11-dihydroxy-5,6,6a,7,8,12b-hexahy-drobenzo[a]rh~n~nth idine hydrochloride, (+)-2 The hydrochloride salt of (+)-5 (217 mg, 0.655 mmol) was dissolved in water and converted to the free base WO96/0~13 2 1 q ~ 2 3 q r~l/~ ~

with conc. NH40H. The free base was extracted into 3 x 25 mL of dichloromethane, the organic solution was dried over MgSO4, filtered, and conc~ntr~te~ to a clear oil using rotary evaporation. The residue was dissolved in 20 mL of dichloromethane and the solution was cooled to -78 ~C. A 1 0 ~ solution of BBr3 in CX~C12 (3 mL, 3 mmol) was added slowly to the solution of the free base via syringe under nitrogen over 30 min. The solution was left to warm to room temperature and was stirred overnight. The reaction was quenched by the addition of 5 mL of methanol. The solvent was removed via rotary evaporation and the flask was left under high vacuum for 8 h. The residue was dissolved in water and the pH was adjusted to 9-10 with a saturated solution of sodium h;~rh~n~te under nitrogen. ~The precipitated free base was ;Rnl~tp~ by suction filtration, washed on the filter with cold water and dried under vacuum. The filtrate was extracted with 3 x 25 mL of dichloromethane. The dichloromethane extracts were dried (MgSO~) filtered and concentrated on the rotary evaporator. This residue and the solid product were combined and dissolved in ethanol.
The solution was acidified with ~th~n~l;c XCl and the volatiles were removed in vacuo. The HCl salt was ~ t~ following cryst~ll;7~ n from EtOAc/isopropanol to yield 158 mg (79~), mp dec >120 ~C; [~] D +83 degrees (c 0.25, EtOH); CIMS (;~h~lt~n~): M+1, 268; 1H NMR (d6 DMSO) ~ 9.62 (s, 2, NH'), 8.87, 8.85 (28 2, OX), 7.43-7.37 (m, 3, ArX), 7.34 (m, 1, ArH), 6.70 (8, 1, ArH), 6.61 (8, 1, ArH), 4.35 (d, 2, ArCH2N, J=3.94 Hz), 4.13 (d, 1, Ar2CH, J = 11.26 Xz), 2.g7-2.90 (m, 1, CHN), 2.78-2.72 (m, 1, ArCH2), 2.72-2.64 (m, 1, ArCHl), 2.18-2.11 (m, 1, CHCN), 1.93-1.84 (m, 1, CXCh'); HRMS (CI, isobutane) Calculated for Cl,Hl3ClNO2 268.1338, Found 268.1332 ~ 096/0~13 2 ~ 9 ~2 3 9 .~

6.1.6.
(6~S,1~ (-)-10,11~l~y~L~ 5,6,6~7,8,1~hnb~y ~ d~1.. Al_ ridine hydrochloride, (-)-2 Following a procedure ;~ntiC~l to that for (+)-2, - s 404 mg ~1.219 mmol) of (-)-5 hydrochloride was converted to 351 mg (94%) of the HCl salt following cryst~ll;7~t;r,n from EtOAc/isu~Luu~llol; mp dec ~120 ~C; [~]D-86 degrees (c 0.25, EtOH); CIMS (i~nhut~n~): M+l, 268; lH NMR (d~
DMSO) ~ 9.61 (5, 2, NH2'), 8.87, 8.85 (28, 2, OH), 7.43-7.37 (m, 3, ArH), 7.34 (m, 1, ArH), 6.70 (s, 1, ArH), 6.61 (8, 1, ArH), 4.35 (d, 2, ArCH2N, J = 3.11 ~z), 4.13 (d, 1, Ar2CH, J = 10.98 Hz), 2.97-2.90 (m, 1, CHN) 2.78-2.72 (m, 1, ArCH2), 2.72-2.64 (m, 1, ArCH2), 2.18-2.11 (m, 1, CHCN), 1.93-1.84 (m, 1, CHCN); HRMS (CI, isobutane) Calculated for Cl7Hl~ClNO2 268.1338, Found 268.1332.

6.1.7.
X-ray Cry~tallogr~phy o$ (6~S,12b)-(-)-N-R-methoxy-phenylacetyl-10,11-di~ethoxy-5,6,7,8,6a,12b-hex~L~l~b~zo[~]rh~"nnt~ i-dine ~(-)-4)]
The absolute confi~uration of this compound was clearly est~h-;~hP~ on the basis of the known (R)-ster~Qrh~m;~try of the O-methyl~-n~l;c acid used to prepare (-)-4.
Crystal data: C28H29NO~; formula weight = 4g3.55;
orthr,rh~ 'ic; space group P212l2l (No. 191; Z - 4, a =

8.8868 (6) A, ~ = 12.027 (4) A, c = 21.940 (2) A, v =
2344 (1) A3; calculated density = 1.26 g/cm3; absorption corfr;r;~nt = 0.78 cm~3. Intensity data were rrll~ctP~
at 20 ~C, with Mo R~ r~ t;r,n (A = 0.71073 A) on an Enraf-Nonius CAD4 computer controlled kappa axis diffractometer equipped with a grap~ite crygtal, ;nri~rnt beam monochromator. Data were roll~rt~ using the -2(theta) scan techni~ue. The scan rate varied from 1 to 20 degrees/min (in omega). The variable scan rate allows rapid data collection for intense reflections where a fast scan rate is used and assures good counting WO 96/02513 ~ 1 q 5~3 9 . ~

statistics for weak reflections where a 310w scan rate is used. Data were collected to a maximum 2 (theta) of 55.0 degrees. The scan range (in deg) was ~t~rm;n~-d as a function of theta to correct for the separation of the doublet (see, CAD4 Operations Manual, E:nraf-Nonius, Delft, 1977). The scan width was calculated as follows:
m scan width = 0.56 + 0.350 tan(theta). Moving-crystal moving-counter background counts were made by scanning an;
additional 25~ above and below this range. Thus, the ratio of peak counting time to background counting time was 2: 1. The counter aperture was also adjusted as a function of theta. The horizontal d~eLLuLe width ranged from 1.9 to 2.4 mm; the vertical aperture was set at 4.0 mm. The diameter of the incident beam rf~ll;r~tr~r was 0.7 mm and the crystal to detector distance was 21 cm. For intense reflections an attenuator was ~l~t~ tically inserted in front of the detector; the attenuator factor was 12.9.
A total of 30i6 reflections were collected, of which 3076 were unique and not ~y~L~ lly absent. Lorentz and polarization corrections were applied to the data.
The linear absorption coefficient is 0.8/cm for Mo Kcl!
radiation. No absorption correction was made. The structure was solved using =S~LX-86 (G.M. Sheldrick, Institut fur ~nr~rg~n; qche Chemie der Universitat Gottingen, F.R.G.) . The L~ ;n;n~ atoms were located in 8~rrP~rl;ng difference Fourier syntheses. Hydrogen atoms were located and added to the structure factor calculations but their positions were not refined. The structure was refined in full-matrix least s~uares where the function m;n;m; 7ed was ~w (¦FO¦ - ¦F~¦) 2 and the weight w is defined as per the Killean and Lawrence method with terms of 0.020 and 1Ø (Killean, R.C.G. and Lawrence, J.~., in Acta Cryst., Sec. B. 1969, 25, 1750-1752.) Scattering factors were taken from Cromer and Waber.
(Cromer, D.T. and Waber, J.T. International Tables for ~ 096/0~13 2 1 9 5 Z 3 9 X-Ray Cry~tallography; The Kynoch Pre~s: sirmingham~
England, 1974; Vol. IV, Table 2.2B.) ~n, 1011 dispersion effects were included in E= (see, Ibers, J.A.
and Hamilton, W.C., in Acta Cryst., 1964, 17, 781-782);
the values for ~f~ and ~f'' were those of Cromer.
(Cromer, D.T.; Waber, J.T., International Ta~le8 for X-Ray Crystallography; The Kynoch Press: Birmingham, England, 1974; Vol. IV, Table 2.3.1.).
Only the 1609 reflections having intensities greater than 3.0 times their standard deviation were used in the r~f-n ~. The final cycle of re~n included 298 variable parameters and converged (largest parameter shift was 0.14 time6 its esd) with unweighted and weighted agreement factors of: Rl = ~IFO - FCl/~Fc =
0.056; R2 = SQRT(~w(Fo - F=)2/~Fo2) = 0.065.
The standard deviation of an observation unit weight was 1.59. There were no correlation coefficients greater than 0.50. The highest peak in the final difference Fourler had a height of 0.21 e/A3 with an estimated error based on ~F (see, Cruickshank, D.W.J., in Acta Cryst.
1949, Z, 154-157) of 0.04. Plots of ~w(¦Fo¦ - IFCI) 2 versus ¦FOII reflection order in data collection, sin theta/lambda, and various classes of indices showed no unusual trends. All calculations were p~Lr~L,-~ on a VAX
computer using SDP/VAS. (Frenz, B.A., in Computing in Cry~tallography, Schenk, H.; Olthof-~A7~ ,, R.; van Konigsveld, H.; Bassi, G.C.; Eds.; Delft University Press: Delft, Holland, 1978; pp 64-71.) 6.2.
Synthe8i~ of Sub8tituted Dihydrex~dine Analog~
The C-2, C-3, and/or C-4-substituted hexahydroben-zo[a]phen~nthridine compounds of the general formula, below, W096/0~13 2 1 9 5 2 3 q 2~ : -R2~4 X~'~ k and pharmaceutically acceptable salts thereof, can:also be separated into their ~ p~nPnt. enan~tiomers by the methods described above, wherein ~r and Hb are trans across ring fusion bond c; R is hydrogen or Cl-C; alkyl;
R~ is hydrogen or a ~henol protecting group; X is fluoro, chloro, bromo, or iodo, or a group of the formula -ORs, wherein Rs i8 hyd~y~ll or a phenol protecting group, provided that when X is a group of the formula -OR~, the groups R1 and Rs can be taken together to form a -CH2-group, thus representing a methylenedioxy functional group bridging the C-10 d C-ll positions on the hexahydrobenzo-[a]ph~n~nthridine ring system (as labeled in the formula abo~e); and R2r R3, and R. are independently selected from the group consisting of hydrogen, Cl-C~ alkyl, phenyl, fluoro, chloro, bromo, iodo, or a group -ORl wherein Rl is as= defined abo~ve, provided that at least one of R" R3, and R~ are other than hydrogen The term "Cl-C~ alkyl" as used herein refers to branched or straight chain alkyl groups comprising one to four carbon atoms, including, but not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl and cyclopropylmethyl.
The term "C1-C~ alkoxy" as used herein refers to branched or straight chain alkyl groups comprising one to four carbon atoms bonded through an oxygen atom, ~ 09610~13 2 1 9 ~ 2 3 9 ~ "~ ~

including, but not limited to, methoxy, ethoxy and t-butoxy.
.

2.1.
2-(N-benzyl-N-4-methylbenzoyl)-6,7-dimethoxy-3,4-dlhy-dro-2-naphthylamine To a solution of 4.015 g ~19.5 mmol) of 6,7-dimethoxy-~-tetralone in 100 mL of toluene was added 2.139 g (1.025 equiv.) of benzylamine. The reaction was heated at reflux overnight under N2 with cnnt;m-ouY water removal. The reaction was cooled and the solvent was removed by rotary vacuum evaporation to yield the crude N-benzyl enamine a_ a brown oil.
Meanwhile, the 4-methylbenzoyl chloride acylating agent was ~L~aL~d by 8nar~n~;ng 3.314 g (24.3 mmol) of p-toluic acid in 200 mL benzene. To this solution waa added 2.0 equiv. (4.25 mL) of oxalyl chloride, dropwise via a pressure-equalizing dropping funnel at 0 ~C. DMF
(2-3 drops) was added to the reaction mixture catalytically and the ice bath was removed. The progress of the reaction was monitored via infrared spectroscopy.
The solvent was removed by rotary vacuum evaporation and the residual oil was pumped down under high vacuum overnight.
The crude N-benzyl enamine residue was dissolved in 100 mL of CH2Cl" and to this solution was added 2.02 g (19.96 mmol) of triethylamine at 0 ~C. 4-methylbenzoyl chloride (3.087 g, 19.96 mmol) was dissolved in 20 mL
CH,Cl2 and this snlutinn was added dropwise to the cold, stirring N-benzyl enamine solution. The reaction was allowed to warm to room temperature and was left to stir under N2 overnight. The reaction mixture was washed successively with 2 x 30 mL of 5~ a~ueous HCl, 2 x 30 mL
of saturated sodium bicarbonate solution, saturated NaCl anlntjnn, and wag dried over MgS0~. After filtration, .35 the filtrate was cnnc~ntrated under vacuum.
Crystallization from diethyl ether gave 5.575 g (69.3%) of the enamide mp 96-98 ~C. CIMS (isobutane); M + 1 414;
H-NMR (CDC13) ~ 7.59 (d, 2, ArH), 7.46 (m, 3, ArH) ! 7.35 (m, 3, ArH), 7.20 (d, 2, ArH), 6.60 (8, 1, ArH), 6.45 (8, 1, ArH~, 6.18 (s, 1, ARCH), 5.01 (o, 2, ArCH2N)! 3.80 (b, 3, OCH3), 3.78 (s, 3, OCH2), 2.53 (t, 2, ArCH,), 2.37 (8, 3, ArCH3), 2.16 (t, 2, CHz); Anal. (C~7H27NO3) C, H, N.

2.2.
Tran~-2-methyl-6-benzyl-10,11-~'' ' y-5,6,6a,7,8, 12b-hexal.yd~b_~zo[a]rh~"onth~idine-5-one A solution of 4.80 g (1 1.62 mmol) of the 6,7-~; t~y enamide preparçd above, in 500 mL of THF, =was introduced to an Ace Glass 500 mL photochemical reactor.
This ~olution was stirred while irrA~; at; n~ for 2 hours with a 450 watt Hanovia medium ~, eSYUL~, ~uartz, mercury-vapor~lamp seated in a water cooled, ~uartz immersion well. The solution was conc~ntnAt~d in vacuo and crystallized from diethyl ether to provide 2.433 (50.7 ~) of the 10,11-dimethoxy lactam, mp 183-195 ~C. CIMS
(isobutane); M + 1 414; IH-NMR (CDCl3) ~ 8.13 (d, 1, ArH), 7.30 (8, 1, ArH), 7.23 (m, 6, ArH), 6.93 (o, 1, ArH), 6.63 (8, 1, ArH), 5.38 (d, 1, ArCH2N), 5.30 (d, 1, ArCH2N), 4.34 (d, 1, Ar2CH, J = 11.4 Hz), 3.89 (~, 3, OCH3), 3.88 (s, 3, OCH3), 3.76 (m, 1, CHN), 2.68 (m, 2, ArCH,), 2.37 (8, 3, ArCH3), 2.25 (m, 1, CH2CN), 1.75 (m, 1, CHlCN); Anal. (C27H2,NO3) C, H, N.

2.3.
Tran~-2-methyl-6-benzyl-10,11-~ ' y-5,6,6a,7,8, 12b-hexaL~d ~L_ zo[~] rh~n~n~h ~idine hydrochloride A solution of 1.349 g (3.27 mmol) o~ the lactam prepared above, in 100 mL dry THF was cooled=in an ice-salt bath and 4.0 e~uiv. (13.0 mL) of 1.0 molar 3H3 was added via syringe.~ The reaction was heated as reflux under nitrogen overnight. Methanol (10 mL) was added dropwise to the reaction mixture and reflux was c~nt;nl1~d for 1 hour. The solvent was removed by rotary vacuum w096~0~3 2 1 9 5 2 3 9 r~
~ 31 evaporation. The residue was chased two times with methanol and twice with ethanol. The flask was placed under high vacuum (0.05 mm Hg) overnight. The residue was dissolved in ethanol and was carefully a~;~if;~ with con~ntrated HCl. The violatiles were removed and the product was crystallized from ethanol to afford 1.123 g (78.9~) of the hydrochloride salt, mp 220-223 ~C. CIMS
(isobutane); M + 1 400; IH-NMR (CrC13, free base) ~ 7.37 (d, 2, ArH), 7.33 (m, 2, ArH), 7.26 (m, 1, ArH), 7.22 (8, 1, ArH), 7.02 (d, 1, ArH), 6.98 (d, 1, ArH), 6.89 (8, 1, ArH)/ 6.72 (8, 1, ArH), 4.02 (d, 1, Ar,CH, J = 10.81 Hz), 3.88 (s, 3, OCH3), 3.86 (d, 1, ArCH2N), 3.82 (m, 1, ArCH2N), 3.78 (s, 3, OCH3), 3.50 (d, 1, ArCH2N), 3.30 (d, 1, ArCH2N), 2.87 (m, 1, ArCH,), 2.82 (m, 1, CHN), 2.34 (m, 1, CH2CN), 2.32 (s, 3, ArCH3), 2.20 (m, 1, ArCH2), 1.93 (m, 1, CH2CN); Anal. (C2~H29NO2)C, H, N.

2.4.
Tran~-2-methyl-10,11-dimethoxy-5,6,6a,7,8,12b-hexahy-drobenzo[a~ r~ . . t 1- ~ ldine hydrochloride A solution of 0.760 g (1.75 mmol) of the 6-benzyl hydrochloride salt prepared above in 100 mL of 95~
ethanol c~nt~in;ng 150 mg of 10~ Pd/C catalyst was shaken at room temperature under 50 psig of H2 for 8 hours.
After removal of the catalyst by filtration through Celite, the solution was concentrated to dryness under vacuum and the residue was recryst~ll; 7~;t from acetonitrile to afford 0.520 g (86.2~) of the crystalline salt, mp 238-239 ~C. CIMS (;q~hnt~n~); M + 1 310; 3HNMR
(~MSO, HCl salt) ~ 10.04 (8, 1, NH), 7.29 (d, 1, ArH), 7.16 (m, 2, ArH), 6.88 (8, 1, ArH), 6.84 (8, 1, ArH), 4.31 (s, 2, ArCH2N), 4.23 (d, 1, Ar,CH, J = 10. 8 Hz), 3.76 (s, 3, OCH3), 3.70 (s, 3, OCH3), 2.91 (m, 2, ArCH2), 2.80 (m, 1, CHN), 2.49 (8, 3, ArCH2), 2.30 (m, 1, CH2CN), 2.09 (m, 1, CH2CN)i Anal. (C20H23NO,) C, H, N.

WO9G~2513 2 1 q52~9 ~ s 2.5.
Tran~-2-mnthyl-10,11-dihydroxy-5,6,6a,7,8,12b--hex~hy-d ob~2~0 [A] rh~n.nfhrldine hydrochloride 0.394 S (1.140 mmol) of the O,O-dimethyl hydrochloride salt prepared above was converted to its free base. The free base was dissolved in 35 mL of dichluL, h~nP and the solution wa3 cooled to -i8 ~C.
4.0 e~uiv. (4.56 mL) of a 1.0 molar solution of BBr3 was added slowly via syringe. The reaction was stirred under N2 overnight with concomitant warming to room temperature. 7.0 mL of me$hanol was added to the reaction mixture and the solvent was removed by rotary vacuum evaporation. The flask was placed under high vacuum (0.05 mm Hg) overnight. The residue was dissolyed in water and was carefully neutralized to its free base initially with sodium bicarbonate and finally with ill~ hydroxide (1-2 drops). The free base was isolated by suction filtration and was washed with cold water. The filtrate was P~r~tPd several times with dichloromethane and the organic extracts were dried, filtered and concentrated. The filter cake and the organic residue were , ' in~, dissolved in ethanol and carefully acidified with concentrated HCl. After removal of the v~ ilPs, the HCI salt was crys~ll;7P~ as a solvate from I h~n~l in a yield of 0.185 g (51 ~), mp (decomposes o 190 ~C). CIMS (i~h~lt~nP); M + 1 282; 3H-NMR (DMSO, HCI salt) ~ 9.52 (s, 1, NH), 8.87 (d, 2, OH), 7.27 (d, 1, ArH), 7.20 (s, 1, ArX), 7.15 (d, 1, ArH), 6.72 (s, 1, ArH), 6.60 (s, 1, ArH), 4.32 (s, 2, ArCH2N), 4.10 (d, 1, ArCH2CH, J = 11.26 Hz), 2.90 (m, 1, CHN), 2.70 (m, 2, ArCH2), 2.32 (s, 3, ArCH3), 2.13 (m, 1, CH2CN), 1.88 (m, 1, CH2CN); Anal. (Cl3HlgNO2) C, H, N.

2 ~ 9523~
~096/0~13 r~

2.6.
2-~-}enzyl-N-3-methy~zoyl)-6,7- -3,4-dihy-drD-2-naphthyl~m1ne To a solution of 3.504 g (1 7.0 mmol) of 6,7-dimethoxy-~--tetralone in 100 mL of toluene was added 1.870 g ~1.025 equiv.) of benzylamine. The reaction was heated at reflux overnight under Nl with ~nnt;nllnus water removal.
The r~ct10~ was cooled and the solvent was removed by rotary vacuum evaporation to yield the crude N-benzyl enamine as a brown oil.
Meanwhile, the 3-methylbenzoyl chloride acylating agent was ~a~ed by sll~r~n~;ng 3.016 g (22.0 mmol) of m-toluic acid in 100 mL benzene. To this solution was added 2.0 equiv. (3.84 mL) of oxalyl chloride, dropwise via a pressure-e~l~'; 7.; ng dropping funnel at 0 ~C. DMF
(2-3 drops) was added to the reaction mixture catalytically and the ice bath was removed. The progress of the reaction was monitored via infrared spectroscopy.
The solvent was removed by rotary vacuum evaporation and the residual oil was pumped down under high vacuum overnight.
The crude N-benzyl enamine residue was dissolved in 100 m~ of CH2Cl2, and to this solution was added 1.763 g (17.42 mmol) of triethylamine at 0 ~C. 3-methylbenzoyl chloride (2.759 g, 17.84 mmol) was dissolved in 20 mL
CH2Cl2 and this solution was added dropwise to the cold, stirring N-benzyl enamine solution. The reaction was allowed to warm to room temperature and was left to stir under N2 overnight. The reaction mixture was washed successively with 2 x 30 mL of 5~ a~ueous HCl, 2 x 30 m~
of saturated sodium bicarbonate solution, saturated NaCl solution, and was dried over MgS0~. After filtration, the filtrate was concentrated under vacuum.
Crystallization from diethyl ether gave 4.431 g (63.1~) of the enamide mp 96-97 ~C. CIMS (isobutane); M + 1 414;
3H-NMR (CDCl3) ~ 7.36 (s, 1, ArH), 7.26 (m, 3, ArH), 7.20 (m, 5, ArH), 6.50 (s, 1, ArH), 6.40 (s, 1, ArH), 6.05 (s, 2l ~23q Wo9610~513 i~

1, ArCH), 4.95 (9, 2, ArCH,N), 3.75 (8, 3, OCH3), 3.74 (8, 3, OCH3), 2.43 (t, 2, ArCH2), 2.28 (8, 3, ArCH3), 2.07 (t, 2, CH2); Anal. (C27H2,NO3) C, H, N.
.
2.7.
Tran~-3-~ethyl-6-b-nzyl-lO,11-~; ' y-5,6,6a,7,8, 12b-hexah~..lL-,L- zo[a]rh~o"onthridine-5-one A Snlllt;nn of 1_922 g (4.65 mmol) of the 6,7-dimethoxy enamide prepared above, in 500 mL of THF, was introduced to an Ace Glass 500 mL photochemical reactor.
This solution was stirred while irr~ t;ng for 5 hours with a 450 watt Hanovia medium pressure, quartz, mercury-vapor lamp seated in a wate:r cooled, ~uartz immersion well. The solution was concentrated in vacuo and crystallized from diethyl ether to provide 0.835 g (43.4%) of the 10, 11 dimethoxy lactam, mp 154-157 ~C.
CIMS (isobutane); M + 1 414; 3H-NMR (CDCl3) ~ 7.94 ~g, 1, ArH), 7.34 (d, 1, ArH), 7.17 (m, 6, ArH), 6.84 (8, 1, ArH), 6.54 (s, 1, ArH), 5.28 (d, 1, ArCH2N), 4.66 (d, 1, ArCH2N), 4.23 (d, 1, Ar,CH, J = 11.4 Hz), 3.78 (8, 3, OCH3), 3.74 (8, 3, OCH3), 3.61 (m, 1, CHN), 2.59 (m, 2, ArCHl), 2.34 (s, 3, ArCH3), 2.15 (m, 1, CH2CN), 1.63 (m, 1, CH2CN); Anal. (C2,H2,NO3) C, H, N.

2.8.
Tranf3-3-~nethyl-6-benzyl-10,11-dimetho~y-5,6,6a,7,8, 12b-hexaL~l-ob_~zo[a]r~ ldinch~ Lloride A solution of 0.773 g (1.872 mmol) of the lactam ~d above, in 50 mL dry THF was cosled in an ice-salt bath and 4.0 equiv. (7.5 mL) of 1.0 molar BH3 ~was added via syringe. The reaction was heated as reflux under ~nitrogen overnight: Methanol (6 mL) was added dropwise to the reaction mixture and reflux wag cnnt;nl1~
for 1 hour. The solvent was removed by rQtary vacuum evaporation. The residue was chased two times with;
methanol and twice with ethanol. The ~las~ was placed under high vacuum (0.05 mm Hg) overnight. The residue 096l0~13 2 1 9 5 2 3 9 . ~11.

was dissolved in ethanol and was carefully acidified with c~n~ntrated HCl. The vi~l~t;1P~ were removed and the product was cryst~ll;7ed from ethanol to afford 0.652 g (80~) of the hydrochloride salt, mp 193-195 ~C. CIMS
(isobutane); M + 1 400; 3H-NMR (CDCl3, free base) ~ 7.38 (d, 2, ArH), 7.33 (m, 2, ArH), 7.28 (m, 2, ArH), 7.07 (d, 1, ArH), 6.90 (s, 1, ArH), 6.88 (8, 1, ArH), 6.72 (8, 1, ArH), 4.02 (d, 1, Ar2CX, J = 11.2 Hz), 3.90 (d, 1, ArCH,N), 3.87 (8, 3, OCH3), 3.82 (m, 1, ArCX2N), 3.78 (s, 3, OCX3), 3.48 (d, 1, ArCH2N), 3.30 (d, 1, ArCH2N), 2.88 (m, 1, ArCH2), 2.82 (m, 1, CHN), 2.36 (m, 1, CH2CN), 2.32 (8, 3, ArCH3), 2.20 (m, 1, ArCX2), 1.95 (m, 1, CX2CN);
Anal. (C27H29NO2) C, H, N.

2.9.
Trans-3-methyl-10,11-dimethoxy-5,6,6a,7,8,12b-hexahy-dl~L_ zo[a]rh~n~n~h~idine hydrochloride A solution of 0.643 g (1.47 mmol) of the 6-benzyl hydrochloride salt prepared above in 100 mL of 95%
ethanol ~nt~;n;ng 130 mg of 10% Pd/C catalyst was shaken at room temperature under 50 psig of H2 for 8 hours.
After removal of the catalyst by filtration through Celite, the solution was concentrated to dryness under vacuum and the residue was recrystallized from acetonitrile to afford 0.397 g (78%) of the crystalline salt, mp 254-256 ~C. CIMS (isobutane); M + 1 310; lHNMR
(DMSO, HCl salt) ~ 10.01 (s, 1, NH), 7.36 (d, 1, ArH), 7.09 (d, 1, ArH), 6.98 (8, 1, ArH), 6.92 (s, 1, ArH), 6.74 (8, 1, ArH), 4.04 (s, 2, ArCX2N), 3.88 (5, 3, OCX3), 3.81 (8, 3, OCX3), 3.76 (d, 1, Ar2CH), 2.89 (m, 2, ArCH2), 2.70 (m, 1, CHN), 2.36 (a~ 3, ArCX3), 2.16 (m, 1, CX2CN), 1.70 (m, 1, CH2CN); Anal. (C20X23NO2) C, H, N.

Wo96102513 2 1 9 5 2 3 9 , ~

2.10.
Tran_-3-methyl-10,11-dihydroxy-5,6,6_,7,8,12b-hexal-yd-~L_-Izo[a]rh~nAn~h~i~n~ hydrochloride 0.520 g (1.51 mmol) of the O,O-dimethyl hydrochloride salt prepared above was converted to its free base. The free base was dissolved in 35 mL of dichloromethane and the solution was cooled to -78 ~C.
4.0 equiv. (6.52 mL) of a 1.0 molar solution of BBr3 was added slowly via syringe. The reaction was stirred under N2 overnight with concomitant warming to room~
temperature. 7.0 mL of methanol was added to the~
reaction mixture and the solvent was removed by rotary vacuum evaporation. The flask was placed under high vacuum (0.05 mm Xg) overnight. The residue was dissolved in water and was carefully neutralized to its free base initially with sodium bicarbonate and finally with ammonium hydroxide (1-2 drops). The free base was isolated by suction filtration and was washed with cold water. The filtrate was extracted several times with dichloromethane and the organic extracts were dried, filtered and ~n~Pntr~tPd. The filter cake and the organlc residue were combined, dissolved in ethanol and carefully acidified with concentrated XCl. After removal of the volatiles, the HC1 salt was cryst~l 1; 7~d as a solvate from methanol in a yield of 0.341 g (71.3~), mp (decomposes ~3 190 ~C). CIMS (;~hnt~nP); M + 1 282; 1H
NMR (DMSO, HC1 salt) ~ 9.55 (8, 1, NH), 8.85 (d, 2, OH), 7.30 (d, 1, ArX), 7.22 (5, 1, ArH), 7.20 (d, 1, ArX), 6.68 (8, 1, ArX), 6.60 (s, 1, ArX), 4.31 (5, 2, ArCX2N), 4.09 (d, 1, ArCX2CH, J = 11.2 Hz), 2.91 (m, 1, CXN), 2.72 (m, 2, ArCX2), 2.35 (s, 3, ArCH3), 2.16 (m, 1, CH2CN, 1.85 (m, 1, CX2CN); Anal (C1BX19NO2) C, X, N.

~ 09610~13 2 1 ~ ~239 2.11.
2-(N-benzyl-N-3-methylkenzoyl)-6,7i -3,4-dihydro-2-naphthylamine To a 301ution of 5.123 g (24.8 mmol) of 6,7-dimethoxy-~--tetralone in 200 mL of toluene was adaed 2.929 g (1.025 equiv.) of benzylamine. The reaction was heated at ~ reflux overnight under N2 with nnnt;nlin-lR water removal.
The reaction was cooled and the solvent was removed by rotary vacuum evaporation to yield the crude N-benzyl enamine as a brown oil.
~Anwh;le, the 2-methylbenzoyl chloride acylating agent was prepared by sncr~n~;nr, 4.750 g ~42.2 mmol) of o-toluic acid in 100 mL benzene. To this solution was added 2.0 equiv. ~7.37 mL) of oxalyl chloride, dropwise via a pressure-~Al; 7inJr dropping funnel at 0 ~C. DMF
~2-3 drops) was added to the reaction mixture catalytically and the ice bath was removed. The progress of the reaction was monitored via infrared spectroscopy.
The solvent was removed by rotary vacuum evaporation and the residual oil was pumped down under high vacuum overnight.
The crude N-benzyl enamine residue was dissolved in 100 m~ of CH2Cl2, and to this solution was added 2.765 g ~1.1 equiv.) of triethylamine at 0 oc. 4-methylbenzoyl chloride (4.226 g, 27.3 mmol) waR dissolved in 25 mL
CH2Cl2 and this solution was added dropwise to the cold, stirring N-benzyl enamine solution. The reaction was allowed to warm to room temperature _nd was left to stir under N2 overnight. The reaction mixture was washed auccessively with 2 x 30 mL of 5~ aqueous HCl, 2 x 30 mL
of saturated sodium birArhnnAte solution, saturated NaCl solution, and was dried over MgS0~. After filtration, the f;1trAte was cnnr~ntrAted under vacuum. The resulting oil was purified via the ~hr~ totron1~tili 7; ng a 5~ ether/dichloromethane eluent mobile phase to yield 3.950 g ~38.5~) of the pure oil. CIMS ~; cnhutAnP); M +
1 414; lH-NMR ~CDCl~) ~ 7.34 (d, 2, ArH), 7.30 (m, 2, WO96/0~13 2 1 ~ 5 2 ~ q ~ C

ArX), 7.25 (d, 2, ArH), 7.14 lm, 2, ArH), 7.07 (m, 1, ArX), 6.47 ~5, 1, ArH), 6.37 ~9, 1, ArX), 6.04 ~5, 1, ARCH), 4.96 ~s, 2, ArCH2N), 3.78 ~8, 3, OCH3), 3.77 ~s, 3, OCX3), 2.39 (s, 3, ArCX3), 2.30 ~t, 2, ArCX3), 1.94 ~t, 2, CHl).

a .12.
Trans-4-methyl-6-benzyl-10,11-dimethoxy-5,6,6n,7, 8,12b-hexaL~d-oL_~zo[a]rhGn~nth idine-5-one A solution of 2.641 g ~6.395 mmol) of the 6,7-dimethoxy enamide prepared above, in 450 mL of THF, was introduced to an Ace Glass 500 mL photochemical reactor.
This solution was stirred while irradiating for 3 hours with a 450 watt Hanovia medium pressure, ~uartz, mercury-vapor lamp seated in a water cooled, guartz immersion well. The solution was concentrated in vacuo and crystallized from diethyl ether to provide 0.368 ~20~ of the 10, 11 dimethoxy lactam, mp 175-176 ~C. CIMS
(isobutane); M + 1 414; 3H-NMR ~CDCl3); ~ 7.88 ~m, 3, Ar~), 7.65 ~d, 1, ArH), 7.40 (m, 2, ArH), 7.21 (m, 2, ArH), 6.87 (8, 1, ArX), 6.60 ~s, 1, ArH), 5.34 (d, 1, ArCHlN), 4.72 ~d, 1, ArCHlN), 4.24 ~d, 1, Ar2CH, ~ = 10.9 Xz), 3.86 ~8, 3, OCX3), 3.85 ~8, 3, OCH3), 3.68 ~m, 1, CHN), 2.73 (5, 3, ArCH3~, 2.64 (m, 2, ArCH,); 2.20 ~m, 1, CH2CN), 1.72 ~m, 1, CH2CN).

2.13.
Tran~3-4-methyl-6-benzyl-10,11-dimethoxy-5,6,6a,7, 8,12b-hexahy~ ~h~n~o~a]rh~n~nth~nrl~d ochloride A solution of 1.640 g ~3.97 mmol) of the lactam prepared above, in 100 mL dry THF was cooled in an ice-salt bath and 4.0 equiv. ~15.9 mL) of 1.0 molar BX3 was added via syringe. The reaction was heated as reflux under nitrogen overnight. Methanol (10 mL) was added dropwise to the reaction mixture and reflux was r~nt;n~led for 1 hour. The solvent was removed by rotary vacuum evaporation. The residue was chased two times with ~ 096l0~13 ~ ~ ~ 5 2 3 q . ~I/U~

methanol and twice with ethanol. The flask was placed under high vacuum (0.05 mm Hg) overnight. The residue was dissolved iA ethanol and was carefully acidified with concentrated HCl. The violatiles were removed and the product was crystAl1;7~d from ethanol to afford 1.288 g (74.5~) of the hydrochloride salt, mp 232-235 ~C. CIMS
(isobutane); M + 1 400; 3H-NMR (CDCl2, free base) ~ 7.38 (d, 2, ArH), 7.33 (m, 2, ArH), 7.27 (d, 1, ArH), 7.24 (m, 1, ArH), 7.16 (m, 1, ArH), 7.06 (d, 1, ArH), 6.85 (8, 1, ArH), 6.71 (8, 1, ArH), 4.05(d, 1, Ar2CH, J = 10.8 Hz), 3.89 (d, 1, ArCH2N), 3.87 (a, 3, OCH3), 3.82(m, 1, ArCH,N), 3.76 (8, 3, OCH3), 3.55 (d, 1, ArCH2N), 3.31 (d, 1, ArCH2N), 2.88 (m, 1, ArCH2), 2.81 (m, 1, CHN), 2.34 (m, 1, CH2CN), 2.20 (m, 1, ArCH2), 2.17 (8j 3, ArCHl), 1.94 (m, 1, CH2CN).

2.14.
Tran~-4-methyl-10,11-dimethoxy-5,6,6a,7,8,12L-hexaL~d ~L _AZO[a] rh~n-nth ridine hydrochloride A solution of 0.401 g (0.92 mmol) of the 6-benzyl hydrochloride salt prepared above in 100 m~ of 95~
ethanol c~nt~;n;ng 100 mg of 10~ Pd/C catalyst was shaken at room temperature under 50 psig of H2 for 8 hours.
After removal of the catalyst by filtration through Celite, the solution was concentrated to dryness under vacuum and the residue was recryst~ d from acetonitrile to afford 0.287 g (90.2~) of the crystalline salt, mp 215-216 ~C. CIMS (isobutane); M + 1 310; 3HNMR
(CDCl3, free base) ~ 9.75 (8, 1, NH), 7.29 (d, 1, ArH), 7.28 (d, 1, ArH), 7.21 (m, 1, ArH), 6.86 (8, 1, ArH), 6.81 (8, 1, ArH), 4.35 (d, 1, ArCH2N), 4.26 (d, 1, ArCH2N), 4.23 (d, 1, Ar2CH, J = 11.17 Hz), 3.75 (8, 3, OCH2), 3.65 (8, 3, OCH3), 2.96 (m, 1, CHN), 2.83 (m, 2, ArCH2), 2.30 (8, 3, ArCH3), 2.21 (m, 1, CH3CN), 1.93 (m, 1, CH2CN) .

WO96/02513 2 1 9 ~ 2 3 q 40 2.15.
Trans-4-methyl-10,11-dlhydroxy-5,6,6a,7,8,12b-hexal-y~L ~zo[a]rh~n~n~hri~n~ hydrochloride 0.485 g (1.40 mmol) of the O,O-dimethyl hydrochloride salt prepared above was converted to its free base. The free base was disEolved in 35 mL of dichlbrometharie and the ~ ;nn was cooled to -78 ~C.
4.0 equiv. (5.52 mL) of a 1.0 molar solution of BBr3 was added slowly via syringe. The reaction was stirred under Ni overnight with concomitant warming to room temperature. 7.0 mL of methanol was added to the reaction mixture and the solvent was removed by rotary vacuum evaporation. The flask was placed under high vacuum (0.05 mm Hg) overnight. The residue was dissolved in water and was cnrefully neutralized to its free base initially with sodium h;~rhnn~te and finally with i hydroxide (1-2 drops). The free base was isolated by suction filtration and was washed with cold water. The filtrate was extracted several times with dichluL th~nP and the organic extracts were dried, filtered and ~ul.cullLL~ted. The filter cake and the organic residue were ;nPd, dissolved in ethanol and carefully acidified with concentrated XCl. After removal of the VnlAt;lP~, the HCl salt was crystallized as a solvate from r--~h~nnl in a yield of 0.364 g (81.6~), mp (~p~ Eer O 195 ~C). CIMS (isobutane); M + 1 282; lH-NMR (DMSO, HCl salt~ ~ 9.55 (s, 1, NH), 8.85 (s, 1, OH), 8.80 (s, 1, OH), 7.28 (m, 2, ArH), 7.20 (d, 1, ArH), 6.65 (s, 1, ArH), 6.60 (s, 1, ArH), 4.32 (d, 1, ArCH2N), 4.26 (d, 1, ArCH2N), 4.13 (d, 1, Ar2CH, J = 11.63 Hz), 2.92 (m, 1, CHN), 2.75 (m, 1, ArCH2), 2.68 (m, 1, ArCH2), 2.29 (s, 3, ArCH3), 2.17 (m, 1, C H2CN), 1.87 (m, 1, CH2CN).
Using the same procedures described above, and those described in U.S. Patent 5,047,536, the compounds set forth in Table 2 below are synthesized.

21 q5~3~
~WO9610~13 r~l,u~-s R
Hb TABLE 2.

ENTRY R Rl R7R3 R~ X
Nln~3ER
1' H H H H E OH
1 H H CHl H H OH
2 H H H CHl H OH
3 H H H H CH~ OH
4 H H C6Hs H H OH
CH~ H CH~ H H OH
6 C~H7 H H CH~ H OH
7 H H C,Hs H H OH
8 H H H C,H~ H OH

9 H H H CHl CH~ Br 11 C,G~ H H CH~ CH~ Br 12 CH~ H H H C7Hs OH
13 C~Hg H H OH H OH
14 H H CHl OH H OH
H H H F H OH
16 H H OH H H Br 17 H H Br H H OH

21 952~9 Wo96~2513 i~

ENTRY R Rl R2 R3 Ri x NUMBER
18 H H H Br H OCH
l9 H H H H Br OCH
H H CH3 Br H OCH
2l CH~ H F H H OH

C2Hs H CHl OH H F
26 C2Hq H CH30 H CH1 F
27 C1H7 H H CH10 H Cl 23 ClH7 H H CHl CH,O Cl 29 C,H7 H C2HcO H H OH

3l C~H9 H CH, H H OH
32 C~Hg H H OH CH~ OH
33 C~H9 H OH Cl H OH
34 C~Hg H Cl OH H OH
C~H7 H H ~ CHl H

The affinity for the ,~Ullldb of Entries l, 2 and 4 for D-l and D-2 binding sites was assayed ~lt~ 1 7~;ng rat brain striatal l- , t~c having D-l and D-2 binding sites labeled with 3H-SCH 23390 and 3H-spiperone, respectively. The data obtained in that assay for dihydrexidine (DHX, Entry l') and the ~ of Entries l, 2 and 4 are reported in Table 3.

2~1 957~39 ~096102513 Y~-~

TABLE 3.

D-l D-2 D-l:D-2 EntriesAffinityAffinity Selectivity 1' (DHX)8 nM 100 nM 13 1 14 nM 650 nM 46 2 7 nM 45 nM 6 4 290 nM 185 nM 0.6 Using the methods outlined in Example 1, dihydrexidine and its substituted analogs are resolved into their respective ~n~nt;: r8, i.e., their (6aR,12bS)-(+)- and (6aS,12bR)-(-)-optical isomers.
Accordingly, the following ~ , including their ~alts (especially their acetic and hydrochloride acid addition salts), 0-alkylated, and N-alkylated analogs, are provided by the methods o~ the pre~ent invention:
(6aR,12bS)-(+)-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydro-benzo[a]rh~n~nthridine;
(6aS,12bR)-(-)-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydro-benzo[a]ph~n~nthridine;
(6aR,12bS)-(+)-2-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]ph~n~nthridine;
(6aS,12bR)-(-)-2-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]ph~n~nthridine;
(6aR,12bS)-(+)-3-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]ph~n~nthridine;
(6aS,12bR)-(-)-3-methyl-10,11-dihydroxy-5~6~6a~7~8~l2b-hexa-hydrobenzo[a]ElhF~n~nthridine;
(6aR,12bS)-(+)-4-methyl-10,11-dihydroxy-5~6~6a~7~8~l2b-hexa-llydLvb~ [a]~h~n~nthridine;
(6aS,12bR)-(-)-4-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]~h~n~nthr;~;nP;
(6aR,12bS)-(+)-2-phenyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]ph~n~nthridine;

WO9~0~13 2 ~ 9 5~3 9 (6aS,12bR)-(-)-2-phenyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-methyl-2-methyl-10,11-dihydroxy-5;6,6a,7,8,12b-hexahydrobenzo[a]ph~n~nthridine;
5(6aS,12bR)-(-)-N-methyl-2-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-propyl-3-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nt~ridine;
(6aS,12bR)-(-)-N-propyl-3-methyl-10,11-dihydroxy-105~6~6a~7~8~l2b-hexahydrobenzo[a]rh~n~nthridinei (6aR,12bS).-(+)-2-ethyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydLvbeLLzo[a]rh~n~nthridine;
(6aS,12bR)-(-)-2-ethyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rh~r~nthridine;
15(6aR,12bS)-(+) -3-ethyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rhPn~nthnidine;
(6aS,12bR)-(-) -3-ethyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rh~on~nthridine;
(6aR~l2bs)-(+?-3~4-dimethyl-lo-bromo-ll-hydr 205,6,6a,7,8,12b-hexahydrobenzo[a]rh~nAnthridine;
(6aS,12bR)-(-)-3,4-dimethyl-10-bromo-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-propyl-2,3-dimethyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexal-ydL~,b~lzu[a]rh~nAnthridïne;
25(6aS,12bR)-(-)-N-propyl-2,3-dimethyl-10,11-dihydrox~y-5,6,6a,7,8,12b-hexal-y-lLub~llzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-ethyl-3,4-dimethyl-10-bromo-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aS,12bR)-(-)-N-ethyl-3,4-dimethyl-10-bromo-11-30hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-methyl-4-ethyl-10,11-dihydroxy-;
5,6,6a,7,8,12b-hexahydrobenzora]rh~n~nthridine;
(6aS,12b~)-(-)-N-methyl-4-ethyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine;
35(6aR,12bS)-(+)-N-butyl-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;

21 q~39 ~096102513 (6aS,12b~) -(-)-N-butyl-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPnAnthridine;
(6aR,12bS)-(+)-2-methyl-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPn~nthridine;
(6aS,12bR)-(-)-2-methyl-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPnAnthridine;
~(6aR,12bS) -(+) -3-fluoro-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rh~n~nthridine;
(6aS,12bR) -(-) -3-fluoro-10,11-dihydroxy-5~6~6a~7~8~12b-hexa-hydLubu--zo[a]rhPn~nthridine;
(6aR,12bS) - (+) -10-bromo-2,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rhPnAnthridirLe;
(6aS,12bR) - (-) -10-bromo-2,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rhPnAnthridine;
(6aR,12bS) - (+) -2-bromo-10, 11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rhPnAnthridine;
(6aS,12bR) - (-) -2-bromo-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]rhPrAnthridine;
(6aR,12bS)-(+)-3-bromo-10-methoxy-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPnAnthridine;
(6aS,12bR)-(-)-3-bromo-10-methoxy-11-hydroxy-5,6,6a,7,8,12b-hexal-y-lLub~ zo[a]rhPnAnthridine;
(6aR,12bS)-(+)-4-bromo-10-methoxy-il-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPnAnthridine;
(6aS,12bR)-(-)-4-bromo-10-methoxy-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPn~nthridine;
(6aR,12bS)-(+)-2-methyl-3-bromo-10-methoxy-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPnAnthridine;
(6as~l2bR)-(-)-2-methyl-3-bromo-lo-meth hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]~hPnAnthridine;
t6aR,12bS)-(+)-N-methyl-2-fluoro-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPn~nthridine;
(6aS,12bR)-(-)-N-methyl-2-fluoro-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-methyl-3-fluoro-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rhPn~nthridine;

- 21 9~239 WO9610~13 P

(6as~l2bR)-(-)-N-methyl-3-fluoro-lo~ll-dihydr 5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n:lnthridine;
~6aR,12bS)-(+)-N-methyl-4-fluoro-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]~h~n~nthridine;
5(6aS,12bR)-(-)-N-methyl-4-fluoro-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-ethyl-10-fluoro-3,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]ph~on~nthridinei (6aS,12bR)-(-)-N-ethyl-10-fluoro-3,11-dihydroxy-105,6,6a,7,8,12b-hexahydrobenzo[a]~h~n~nthridine;
(6aR,12bS)-(+)-N-ethyl-2-methyl-10-fluoro-3,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phon~nthridine;
~6aS,12bR)-~-)-N-ethyl-2-methyl-10-fluoro-3,11-dihydroxy-5,6,6a,7,8,12b-hexahydL~b~~ [a]rh~n~nthridine;
15(6aR,12bS)-(+)-N-ethyl-2-methoxy-4-methyl-10-fluoro-ll-hydrcxy-5l6l6al7l8ll2b-hexahydrobenzo[a]phenanth ridine;
(6aS,12bR)-(-)-N-ethyl-2-methoxy-4-methyl-10-fluoro-;
11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanth-ridine;
(6aR,12bS)-(+)-N-propyl-3-methoxy-10-chloro-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]ph~n~nthridine;
(6aS,12bR)-(-)-N-propyl-3-methoxy-10-chloro-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]ph~n~nthridine;
25(6aR,12bS)-(+)-N-propyl-4-methoxy-3-methyl-lo-chloro-ll-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phen- .
anthridine;
(6aS,12bR)-(-)-N-propyl-4-methoxy-3-methyl-10-chloro-11-hydroxy-5,6,6a,7,8,12b-hexahydLub~nz~[a]phen-30anthridine;
(6aR~l2bs)-(+)-N-propyl-2-ethoxy-lo~ll-dihydr 5,6,6a,7,8,12b-hexahydrobenzo[a]ph~n~nthridine;
(6aS,12bR)-(-)-N-propyl-2-ethoxy-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phPn~n~hridinei 35(6aR,12bS)-(+)-N-propyl-4,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]ph~nAnthridine;

~09610~13 21 952~9 r 1ll ~

(6aS,12bR)-(-)-N-propyl-4,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~-n~nthridine;
(6aR,12bS)-(+)-N-butyl-2-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]ph-~n~nthridine;
(6aS,12bR)-(-)-N-butyl-2-methyl-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-butyl-4-methyl-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aS,12bR)-(-)-N-butyl-4-methyl-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-butyl-3-chloro-2,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrub~llzo[a]rh~n~nthridine;
(6aS,12bR)-(-)-N-butyl-3-chloro-2,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~nthridine;
(6aR,12bS)-(+)-N-butyl-2-chloro-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]ph~n~nthridine;
(6aS,12bR)-(-)-N-butyl-2-chloro-3,10,11-trihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]~h~n~nthridine;
(6aR,12bS)-(+)-N-butyl-3-methyl-10-iodo-11-hydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]rh~n~rthridine;
(6aS,12bR)-(-)-N-butyl-3-methyl-10-iodo-11-hydroxy-5,6,6a,7,8,12b-hexahydlub~llGu[a]rh.on~nthridine.

6.3.
F' l~gY
6.3.1.
Methods [3H]-SCH23390 was synth~;7ed as described by Wyrick, S. et al., in ~. ~abel. Comp. Radiopharm. 1986, 23, 685-692. (i)-2 was synthesized as previously described. [3H]-Spiperone was purchased from Amersham Corp (Arlington Heights, IL). Nal2sI was supplied by New England Nuclear (Boston, MA), and HEPES buffer was purchased from Research Organics, Inc. (Cleveland, OH).
SCH23390 was a gift from Schering Corp. (Bloomfield, NJ) or was purchased from Research Biochemicals Inc. (Natick, 219523~ ~
WO 96/02513 r MA). Domperidone and ketanserin were gifts of Janssen Phar~-cff~ltir~ (New Brunswick, NJ). Dopamine, cAMP, isobutyl methyl~nth;nf~ (IBMX), and chloL~L~ 7ine were obtained from Sigma Chemical Co (St. Louis, ~.O). cAMP
primary antibody was obtained from Dr. Gary Brooker (George Waefhington University, Washington D.C.), and secondary antibody (rabbit anti-goat IgG) covalently attached to magnetic beads was purchased from Advanced Magnetics, Inc. (Cambridge, MA).

6.3.2.
Dl ~nd D, Rafdlf~cc~to~ Assays Radioligand binding followed the method of Schulz et al. (See, Schulz, D.W. et al., in fJ. Neurochem. 1985, 4~, 1601-1611) with minor modifications. For the rat sftudies, male Sprague-Dawley rats (Charles River, Raleigh, NC) weighing 200-400 g were decapitated, and the brains f~uickly removed and placed into ice cold saline.
After a brief chilling period, brains were sliced into 1.2 mm coronal slices with the aid of a dissecting block similar to that described by Heffner, T.G. et al., in ph~frr-f~n7, ~lochem. Behav. 1980, 13, 453-456. The striatum was dissected from two slices cnnff-~in;ng the majority of this region, and the tisfsue was either used ; ~fAtPly or stored at -70 ~C until the day of the assay. After dissection/ rat striata were homogenized by seven manual strokes in a Wheaton Teflon-glass ~I J ;7f~r with ice cold 50 mM HEPES buffer with 4.0 mM
MgCll, pH 7.4 (25 ~C). Tissue wa9 centrifuged at 27,000 x g (Sorvall RC-5B/SS-34 rotor, DuPont, Wilmington DE) for 10 min, and the supernatant was discarded. The pellet was homogenized (fivf- strokes) anfl resflqpf~nflf-d in ice cold buffer and centrifuged again. The final pellet;
was suspended at a concentration of approximately 2.0 mg wet weight/mL. Assay tubes (1 mL final volume) were ;nnnh~tf-f~ at 37 ~C for 15 minutes. ~flnnqpf~f~ffic binding WO96/025l3 ~l 9 5 ~ 3 ~ r~

of [3H]-SCH2339Q (ca. o.25 nM) was defined by adding unlabeled SCH23390 ~1 ~M). Binding was t~rm;nAtPd by filtering with 15 mL ice cold buffer o~ a Skatron or Brandel cell harvester ~Skatron Inc., Sterling, VA;
Brandel Inc., Gaithersburg, MD) using glass fiber filter mats (Skatron no. 7034; Brandel GF/B).
Filters were allowed to dry, and 2-4 mL of Scintiverse E (Fischer Sr;~nt;f;r Co., Eair Lawn, NJ) was added. After shaking for 30 min, radioactivity was determined on an LKB-1219 Betarack liquid sr;nt;llAtion counter. Tissue protein levels were estimated using the Folin reagent method of Lowry, O.H. et al., in J. Biol.
Chem. 1951, 193, 265-275, adapted to a Technicon Autoanalyzer I ~Tarrytown, NY).
For D,-like receptors, the pL~ceduLe was as described for Dl-like receptors with the following changes. [3H]-Spiperone was used as the radioligand, and non-specific binding of [3H]-spiperone was defined by adding unlabeled 1 ~M chl~L~, 7;n~ Ketanserin ~50 nM) was added to mask binding of [3H]-spiperone to serotonin receptors.

6.3.3.
Adenylate Cyclase Men~ t~: Striat~
The automated HPLC method of Schulz and Mailman (see, Schulz, D.W. and Mailman, R.B., in ~. Neurochem.
1984, 42, 764-774) was used to measure adenylate cyclase activity by separating cAMP from other labeled nucleotides. Briefly, rat striatal tissue was removed and homogenized at 50 mL/g tissue in 5 mM HEPES buffer (pH 7.5) rrntAin;nr~ 2 mM EGTA. After hcmog~nl7~t;r~n with a Teflon-glass homogenizer, 50 mL/g of 100 mM HEPES-2 mM
EGTA was added and mixed with one additional stroke. A
20 ~L aliquot of this tissue homogenate was added to a ~Le~aLed reaction mixture, yielding a final volume of 100 ~L rr,nt~;n;nJr 0.5 mM ATP, 0.5 mM IBMX, [~32P]-ATP (0.5 w09~0~l3 2 1 q 5 2 3 q ~Ci), 1 mM cAMP, 2 mM MgCl~, 100 mM HEPES buffer, 2 ~M
GTP, O or 100 ~M ~p~;n~ and/or drug, 10 = mM
ph~9Fh~eatine and 5U creatine ph~rh~k;n~e The reaction was initiated by transferring the samples from an ice bath to a water bath at 30 ~C and terminated 16 minute3 later by the addition of 100 ~ of 3~ sodium dodecyl sulfate. Proteins and much of the non-cyclic nucleotides were precipitated by the addition of ioo ~
each of 4.5~ ZnSO, and 10~ Ba~OH), to each incubation tube. The sample3 wer~ centrifuged at 10,000 g for 8 minutes, and the aupernatants removed and loaded onto an ISIS Autoinjector. ~ ~
The HPBC separations were ~r;~ lt using a Waters RCM 8 x 10 module equipped with a C18, lO ~m cartridge, using a mobile phase of 150 mM sodium acetate, 24~
methanol, pH 5Ø A flow rate of 1.3 mL/min was used for separation. A UV detector~set at 254 nm was used to measure the nnl~h~led cAMP, which was added to the sample tubes to serve as an internal standard and as a marker for the labeled cAMP. Sample recovery was based on av mea~uL~ of total unlabeled cAMP peak areas. The r~ t;vity in each iraction was determined by an on-line HPLC rA~1~t;nn detecto~ (Inus Systems, Tampa FL).

6.3.4 andiu~ccc~tol A##ay# in Tran~foctcd Dl acc~t~
The present studies were c~n~n~t~ with Ltk- cells (mouse fibroblasts) that expressed the human receptor, L-hD1. (Biu, Y.F. et al., in ~ol. Endocrinol.
1992, 6, 1815-1824.) The cells were grown in DMEM-H
medium c~nt~;n;ng 4,500 mg/L glucose, L-glutamine, _10 fetal bovine serum and 700 ng/m.~ G418. In these studies, D1 receptor levels were ca 5,000 fmol/mg protein.~All~
cells were m-;nt~;n~ in a hllm;~;f;~ incubator at 37 ~C
with 5~ CO,. Cells were grown in 75 cm' flasks until confluent.

~ 096/02513 2 1 q ~ 2 3 9 The cells were rinsed and lysed with 10 mL of ice cold hypoosmotic buffer (HOB) (5 mM HEPES, 2.5 mM MgCl2, 1 mM EDTA; pH 7.4) for 10 minutes at 4 ~C. Cells were then scraped from the flasks using a sterile cell scraper from Baxter (McGaw Park, IL). Fla8ks received a final rinse with 5 mL of HOB. The final volume of the cell Sncp~nc;~n recovered from each flask was ca. 14 mL.
Scraped membranes from several flasks were then ~c ~;n~.
The ~ ';ne~ cell suspension was homogenized (10 strokes), 14 mL at a time, using a 15 mL Wheaton Teflon-glass homogenizer. The cell homogenates were combined and spun at 43,000 x g (Sorvall RC-5B/SS-34, DuPont, Wilmington, DE) at 4 ~C for 20 minutes. The sup~rnct~nt was removed, and the pellet was resllcpPn~ed (10 strokes) in 1 mL of ice cold HOB for each original flask of cells homogenized. This ' -, te was then spun again at 43,000 x g at 4 ~C for 20 minutes. The supernatant was removed and the final pellet was resuspended (10 strokes) in ice cold storage buffer (50 mM HEPES, 6 mM MgCl" 1 mM
EDTA; pH 7.4) to yield a final concentration of ca. 2.0 mg of protein/mL. Aliquots of the final h~ te were stored in microcentrifuge tubes at -80 ~C.
Prior to their use for radioligand binding or adenylate cyclase assays, protein levels for each membrane preparation were quantified using the BCA
protein assay reagent (Pierce, Rockford, IL) adapted for use with a microplate reader (Molecular Devices; Menlo Park, CA).
Frozen membranes were thawed and resuspended in assay buffer (50 mM HEPES with 6 mM MgCl2 and 1 mM EDTA;
pH 7 4) ~nt~;n;ng a fixed concentration of [3H]SCH23390 (0.2 nM) in a final assay volume of 500 L. Triplicate determinations were performed at data point. Assay tubes were incubated at 37 ~C for 15 min. Tubes were filtered rapidly through Skatron glass fiber filter mats, and the filters rinsed with 5 mL of ice-cold assay buffer using W096/025~3 ~1 9 5 ~ 3 q ~ r ~

a Skatron Micro Cell Harvester (Skatron Instruments Inc., Sterling, VA). Filters were allowed to dry, then punched into sn;nt~ t;nn vials (Skatron In~LLI ts Inc., Sterling, VA). OptiPhase 'HiSafe' II grint;llAt;nn cocktail (2 mB) waa added to each vial. After shaking for 30 min, raaioactivity in each sample was det~Yrin on an B~B Wallac 1219 Rackbeta liguid $n;nt;ll~tion counter tWallac Inc., Gaithersburg, MD).
The foregoing examples of preferred embo~; ~ are provided simply to illustrate the present invention.
other embodiments of the present invention are apparent to one of ordinary skill in the art and are considered to fall within the scope and spirit of the present invention. Hence, the examples are not to be cons~ruea to limit the invention in any way, which invention is limited solely by the claims that follow.

Claims (25)

WHAT IS CLAIMED IS:

1. A pure optical isomer of a compound of the formula and pharmaceutically acceptable salts thereof wherein the Ha and Hb are trans across ring fusion bond c, R is hydrogen or C1 - C4 alkyl R1 is hydrogen or a phenol protecting group X is fluoro, chloro, bromo, iodo, or a group of the formula -OR5 wherein R5 is hydrogen or a phenol protecting group, provided that when X is a group of the formula -OR5, the groups R1 and R5 can be taken together to form a group of the formula -CH2-; and R2, R3 and R4 are independently selected from the group consisting of hydrogen, C1-C4 alkyl, phenyl, fluoro, chloro, bromo, iodo, or a group -OR1 wherein R1 is as defined above, provided said compound is optically active; with the proviso that when R is H, CH3 or n-C3H7, at least one of R1-R4 are not H.

2. The compound of claim 1 which is the (+)-isomer.

3. The compound of claim 1 which is the (-)-isomer.

4. The compound of claim 1 in which the groups R2, R3, and R4 are all hydrogen and R1 is not H.

5. The compound of claim 1 in which at least one of the groups R2, R3, and R4 is methyl.

6. The compound of claim 1 wherein X is hydroxy and R is hydrogen or C1-C4 alkyl.

7. The compound of claim 1 wherein X is selected from the group consisting of fluoro, chloro, bromo, or iodo and R is hydrogen, methyl or n-propyl.

8. (6aR,12bS)-(+)-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]phenanthridine, an O-alkylated or N-alkylated analog thereof, or its pharmaceutically acceptable acid addition salt.

9. (6aS,12bR)-(-)-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]phenanthridine, an O-alkylated or N-alkylated analog thereof, or its pharmaceutically acceptable acid addition salt.

10. (6aR,12bS)-(+)-10,11-dimethozy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]phenanthridine, or its pharmaceutically acceptable acid addition salt.

11. (6aS,12bR)-(-)-10,11-dimethoxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]phenanthridine, or its pharmaceutically acceptable acid addition salt.

12. (6aR,12bS)-(+)-6-(R-methoxyphenylacetyl)-10,11-dimethoxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine, or its pharmaceutically acceptable acid addition salt.

13. (6aS,12bR)-(-)-6-(R-methoxyphenylacetyl)-10,11-dimethoxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine, or its pharmaceutically acceptable acid addition salt.

14. Use of a compound of claim 1 or 4 for the preparation of a medicament for treating a dopamine-related dysfunction of the central nervous system evidenced by an apparent neurological, psychological, physiological, or behavioral disorder.

15. A pharmaceutical composition for treating dopamine-related dysfunction of the central nervous system by an apparent neurological, physiological, psychological, or behavioral disorder, said composition comprising a therapeutically effective amount of the compound according to claim 1 or 4 and a pharmaceutically acceptable carrier therefor.

16. A pharmaceutical composition comprising an effective amount of (6aR,12bS)-(+)-10,11-dihydroxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]phenanthridine, an O-alkylated or N-alkylated analog thereof or its pharmaceutically acceptable acid addition salt and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising an effective amount of (6aR,12bS)-(+)-10,11-dimethoxy-5,6,6a,7,8,12b-hexa-hydrobenzo[a]phenanthridine, an N-alkylated analog thereof or its pharmaceutically acceptable acid addition salt, and a pharmaceutically acceptable carrier.

18. A method of resolving racemic 10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine comprising:
(a) providing a 10,11-diprotected analog of 10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine;
(b) allowing said 10,11-diprotected analog to react with a chiral auxiliary ligand that is a homochiral alpha-alkoxyphenylacetyl halide to provide a pair of diastereomers;
(c) separating said pair of diastereomers into its component stereoisomers to provide a resolution of said racemic 10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine.

19. The method of claim 18 which further comprises removing said chiral auxiliary ligand from one of said component stereoisomers.

20. The method of claim 19 which further comprises regenerating said 10,11-dihydroxy-5,6,6a,7,8,12b-hexahydro-benzo[a]phenanthridine from its 10,11-diprotected analog, said 10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine being optically active.

21. Crystalline (6aR,12bS)-(+)-10,11-dihydroxy-5,6,6a,7,8, 12b-hexahydrobenzo[a]phenanthridine as its pharmaceutically acceptable acid addition salt.

22. The compound of claim 21 which is the hydrochloride.

23. Use of a compound of claim 8 for the preparation of a medicament for alleviating the effects of Parkinson's disease, CNS movement-related disorder, or cardiovascular disorder.

24. Use of a compound of claim 8 for the preparation of a medicament for enhancing endocrine function.

25. A pure optical isomer of a compound of the formula and pharmaceutically acceptable salts thereof wherein the Ha and Hb are trans across ring fusion bond c, R is hydrogen or C1 - C4 alkyl R1 is hydrogen or a phenol protecting group X is fluoro, chloro, bromo, iodo, or a group of the formula -OR5 wherein R5 is hydrogen or a phenol protecting group, provided that when X is a group of the formula -OR5, the groups R1 and R5 can be taken together to form a group of the formula -CH2-; and R2, R3 and R4 are independently selected from the group consisting of hydrogen, C1-C4 alkyl, phenyl, fluoro, chloro, bromo, iodo, or a group -OR1 wherein R1 is as defined above, provided said compound is optically active; with the proviso that at least one of the groups R2, R3, and R4 is methyl.