AU679635B2 - Method for treating 5HT-2B receptor related conditions - Google Patents

Description

WO 95/24200 PCT/US95/03099 1

TITLE

METHOD FOR TREATING 5HT2B RECEPTOR RELATED CONDITIONS Field of the Invention The present invention relates to a method for treating 5HT2B receptor related conditions. Further, this application discloses new compounds of Formulas XI and XII infra.

Backaround of the Invention This invention is directed to a method for treating a mammal suffering from or susceptible to a condition associated with modulation of a 5-HT2B receptor.

Blocking serotonin receptors has been shown to result in 1 number of beneficial pharmacological effects, including reduction in disease states such as hypertension, depression, anxiety, and the like; see U.S. Patent No.

5,141,944. Nelson et al., Psvchopharmacolocv and Biochemistry of Neurotransmitter Receptors, eds. H.I.

Yamamura et al., Elsevier/North Holland Inc., p 325, have confirmed that there are multiple serotonin recognition sites. The general class of serotonin receptors are referred to as the 5-HT receptors.

WO 95/24200 PCT/US95/03099 2 Specific 5-HT receptor sites include 5-HTIA, 5-HTiB, 5-HT1D, 5-HT2A, 5-HT2B, 5-HT2C, 5-HT 3 and 5-HT4 sites.Each of these receptors mediates certain physiological effects. See Leonard, International Clinical PsychoDharmacolovy, 7:13-21 (1992).

This invention provides a method for using compounds which are active at the 5-HT2B receptor to trFit or prevent 5-HT2B related conditions. Further, this invention provides a method for selectively blocking the 5-HT2B receptor. Additionally, this invention provides a method for blocking human 5-HT2B receptors. The 5-HT2B receptor active compounds provide a useful tool for characterizing the 5-HT2B receptor.

This invention provides a group of compounds which are 5HT2B receptor antagonists. Applicants have discovered that such compounds are potent competitive inhibitors of serotonin-induced contraction of the colon. Thus, this invention provides compounds which can act to normalize gastrointestinal motility and be useful in the treatment of Functional Bowel Disorders.

Further, it has been discovered the 5-HT2B receptor is lo-alized in the rat lung, stomach fundus, uterus, bladaer, and colon. Interesting areas of 5-HT2B receptor localization in the human include but are not limited to the brain and blood vessels. Thus, conditions which can be treated using a compound which modulates a 5-HT2B receptor includes, for example, psychosis, depression, anxiety disorders, uterine diseases such as endometriosis, fibrosis, and other abnormal uterine contractivity, panic attack, migraine, eating disorders, seasonal affective disorder, consumption disorders, cardiovascular conditions, such as thrombosis, hypertension, angina, vasospasm, and other vascular occlusive diseases, incontinence, bladder dysfunction, respiratory/airway disorders including asthma, and the like.

WO 95/24200 PCT/US95/03099 3 Summary of the Invention This invention provides a method for treating a mammal suffering from or susceptible to a condition associated with dysfunctional or abnormal 5-HT2B receptor stimulation, comprising administering an effective amount of a compound interacting with the 5HT2B receptor as an agonist, partial agonist or antagonist selected from the group consisting of a compound of the Formula I

AI

IA NR 1

I

Ra Q wherein: Q is hydrogen or (CHR 2 )R4 R1 is hydrogen or CI-C 3 alkyl; R2 is hydrogen or C 1

-C

3 alkyl; R3 is hydrogen or Ci-C 3 alkyl; R4 is C5-C8 cycloalkyl, substituted C 5

-C

8 cycloalkyl, Cs-

C

8 cycloalkenyl, substituted C 5

-C

8 cycloalkenyl, bicyclic or substituted bicyclic; A is selected from the group consisting of

R

6 R79C I I a)

RE

RE

LI (IIIa) and

IILBR

WO 95/24200 WO 9524200PCTUS95/03099 4

R

6 where in R6 and R 7 are, independently, hydrogen, CI-C 6 alkyl,

C

2

-C

6 alkenyl, halo, halo (Cl-C6) alkyl, halo (C 2

-C

6 alkenyl, CI-Clo alkanoyl, CO 2 R5., (Cl-C 6 alkyl)mamino, N0 2

-SR

5 or mn is 1 or 2; is independently hydrogen or C 1

-C

4 alkyl; is C 1

-C

4 alkyl; R8 is independently selected from the group consisting of an R6 group, substituted C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl- (Cl-C 3 alkyl, CS-C 8 cycloalkenyl, substituted Cs-C 8 cycloalkenyl, Cs-C 8 cycloalkenyl-(Cl-

C

3 )alkyl, C 7

-C

2 0 arylalkyl; or R6 and R7 together with the carbon atoms of group A form a 5- to 8.-member carbon ring;, a compound of Formula 11

R

8 R12' N

N

1 4 i

R

11

I

H

-wherein R8 is selected from the group consistinq of hydrogen, Cl-C 6 alkyl, C 2

-C

6 alkenyl, halo, halo (C2--C6)alkyl, halo (C 1

-C

6 alkenyl, COR5, Cj-C.O calkanoyl, CO 2 Rs', (Cl-C 6 aly~aio 0,-SR5, OR5, substituted C 3 -CS cycloalkyl, C3-

C

8 cycloalkyl, C 3 -CS cycloalkyl-(C 1

-C

3 )alkyl, C 5

-C

8 cycloalkenyl, substituted Cs-C 8 cycloalkenyl, Cs-C 8 cycloalkenyl- (Cl-C 3 alkyl, and C 7

-C

2 o arylalkyl; is independently hydrogen or C 1

-C

4 alkyl; is C 1

-C

4 alkyl; WO 95/24200 WO 9524200PCTIUS95/03099 R9 and R 10 are independently selected from the group consisting of hydrogen, Cl-C 6 alkyl, substituted C 3

-C

8 cycloalkyl, C 3

-C

8 cycloaJlkyl, C 3

-C

8 cycloalkyl- (CI-C 3 alkyl,

C

5

-C

8 cycloalkenyl- (Ci-C 3 alkyl, C 7

-C

2 0 arylalkyl; R11 is selected from the group consisting Of Cl-C 4 alkyl,

OR

5 fluoro, bromo, iodo, and chloro; R12' is selected from the group consisting of hydrogen and CI-C 4 aZlkyl; a compound of Formula III 0

CR

13

R'

NR 12

N

wherein:

R

12 is C 1

-C

4 alkyl or allyl;

R

1 3 is or

R

15 is hydrogen or Cl-C 4 alkyl;

R

1 4 is C 1

-C

4 alkyl, hydroxy Cl-C 4 alkyl, C 3

-C

7 cycloalkyl, and C 3

-C

7 cycloalkyl substituted with hydroxy or methoxy; a compound of Formula IV WO 95/24200 WO 954200PJUS95/03099 wherein:

R

15 is C 1

-C

4 alkyl;

R

1 6 is allyl or CJ.-C4 straight chain alkyl;

R

17 is hydrogen or CI-C 4 straight chain alkyl;

R

18 is hydrogen, Cl-C 4 alkyl, hydroxy, or CI-C 4 alkyloxy; is 0, 1, 2, or 3; a compound of Formula V 0o 2 1I 1 2 C-N -(alk)-R wherein:

R

19 is C 1

-C

4 alkyl;

R

20 is allyl or C 1

-C

4 straight chain alkyl; WO 95/24200 WO 9524200PCT[US95/03099 7

R

2 1 is hydrogen or C 2

C

4 straight chain alkyl;

R

2 2 is pyridinyl or imidazolyl; alk is a divalent organic radical derived from a straight or branched Cl-Cs alkane; a compound of Formula VI CHR 24 -R R 2 1 wherein:

R

23 is C 1

-C

3 alkyl or allyl;

R

2 4 is C 1

-C

3 hydroxyalkyl. or CI-C3 dihydroxyalkyl;

R

2 5 is hydrogen or CH 3 a compound of Formula VII WO 95/24200 WO 9524200PCT[US95/03099 8 a compound of Formula VIII

H

N

N

lz v

R

25 is hydrogen or methoxy; a compound of Formula IX wherein: Yb, in combination with the carbon atom to which it is joined, defines a substituted or unsubtituted aromatic heterocyclic 5-membered ring selected from the group consisting of

N-.

0

R

29 0 N R 29 -CH,

R

2 6 is hydrogen, Cl-C 3 alkyl, allyl, or

I

WO 95/24200 PCT/US95/03099 9

R

27 is hydrogen, CI-C 3 alkyl, allyl, or (CH2)n'-X"; n' is 1 to X" is an optionally substituted phenyl, CI-C 3 alkoxy, or C1-C3 alkylthio;

R

2 8 and R 2 9 are independently hydrogen, C1-C3 alkyl, Ci- C3 alkox-, hydroxy, C1-C3 alkylthio, halo, CN, phenyl; or together are p" is 3 to 6; ya is -CH 2 m" is 0, 1, or 2; and a compound of the Formula X

N

N

or a pharmaceutically acceptable salt or solvate thereof.

This invention provides a method for treating a mammal suffering from or susceptible to a condition associated with dysfunctional or abnormal 5-HT2B receptor stimulation, comprising administering an effective amount of a compound interacting with the 5HT2B receptor as an agonist, partial agonist or antagonist selected from the group consisting of a compound of the Formula XI

-I

WO 95/24200 WO 9524200PCJ'1US95/03099

R

30

R

3 1 r s~NRl xi

R

3 Qf where in Q' is selected from the group consisting of hydrogen, R 3 4 and

(CHR

2

R

4

R

34 is selected from the group consisting of spirobicyclic, substituted spix-o-bicyclic, bicyclic or substituted bicyclic; RI is hydrogen or C 1

-C

3 alkyl;

R

2 is hydrogen or Cl-C 6 alkyl;

R

3 is hydrogen Or C 1

-C

3 alkyl;

R

4 is C5-C 8 cycloalkyl, substituted C5-C 8 cycloalkyl,

C

8 cycloalkenyl, substituted C 5

-C

8 cycloalkenyl, bicyclic or substituted bicyclic; A is gelected from the group consisting of Ra

R

6 R8 ,an WO 95/24200 PCT/US95/03099 11 wherein

R

6 and R 7 are, independently, hydrogen, Ci-C 6 alkyl,

C

2

-C

6 alkenyl, halo, halo(C 1

-C

6 )alkyl, halo(C 2

-C

6 )alkenyl,

COR

5

C

1

-C

10 alkanoyl, C0 2 R5', (Ci-C6 alkyl)mamino, NO 2

-SR

5 or OR 5 m is 1 or 2;

R

5 is independently hydrogen or CI-C 4 alkyl;

R

5 is CI-C 4 alkyl;

R

8 is independently selected- from the group consisting of an R 6 group, substituted C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl-(C 1

-C

3 alkyl, Cs-C 8 cycloalkenyl, substituted Cs-C 8 cycloalkenyl, Cs-CS cycloalkenyl-(Ci-

C

3 )alkyl, C 7

-C

2 0 arylalkyl; or

R

6 and R 7 together with the carbon atoms of group A form a 5- to 8-member carbon ring;

R

3 0 and R 3 1 join to form a 3 to 8 member carbon ring; or

R

3 0 and R-1 are independently selected from the group consisting of Ci-C6 alkyl and C 2

-C

6 alkenyl; or a pharmaceutically acceptable salt or solvate thereof.

This invention provides a method for treating a mammal suffering from or susceptible to a condition associated with dysfunctional or abnormal 5-HT2B receptor stimulation, comprising administering an effective amount of a compound interacting with the 5HT2B receptor as an agonist, partial agonist or antagonist selected from the group consisting of a compound of the Formula XII

R

3 0 R 3 1

A

A NR 9

R

1

I

H

XII

A is selected from the group consisting of Y II I_ I WO 95/24200 WO 95/4200 CT1US95/03099 12

R,

R7 (Ia) Ran

R

6 R7 (Iva) where in R6 and R 7 are, independently, hydrogen, C 1

-C

6 alkyl,

C

2

-C

6 alkenyl1, halo, halo (Cl-C6) alkyl, halo (C 2

-C

6 alkenyl,

C

1

-C

10 O alkanoyl, CO 2

R

5 (Cl-C 6 alkyl)mamnino, NO 2

-SR

5 or OR 5 mn is 1 or 2;

R

8 is selected from the group consisting of hydrogen, C 1

-C

6 alkyl, C 2

-"C

6 alkenyl, halo, halo (C 2

-C

6 alkyl, halo (Cl-C6) alkenyl, C0R 5

C

1 -Cj 0 alkanoyl, CO 2 RS', (Cl-C 6 alkyl)mamino, NO 2 -SR5, OR 5 substituted C 3 -C8 cycloalkyl, C 3

C

8 cycloalkyl, C 3

-C

8 cycloalkyl-(C 1

-C

3 )alkyl, C 5

-C

8 cycloalkenyl, substituted C 5 -C8 cycloalkenyl, Cs-Cs cycloalkenyl-(Cl-C 3 )alkyl, and C 7

-C

2 0 arylalkyl; is independently hydrogen or C 1

-C

4 alkyl; is CI-C 4 alkyl;

R

6 and R 7 together with the carbon atoms of group A form a 5- to 8-member carbon ring;

R

9 and RIO are independently selected from the group consisting of hydrogen, Cl-C 6 alkyl, substituted C 3

-C

8 WO 95/24200 PCT/US95/03099 13 cycloalkyl, C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl-(Ci-C3)alkyl,

C

5

-C

8 cycloalkenyl-(Cl-C 3 )alkyl, C7-C 20 arylalkyl; R11 is selected from the group consisting of C 1

-C

4 alkyl, ORs5, fluoro, bromo, iodo, and chloro;

R

3 0 and R 3 1 join to form a 3 to 8 member carbon ring; or

R

3 0 and R 3 1 are independently selected from the group consisting of C1-C6 alkyl and C 2

-C

6 alkenyl; or a pharmaceutically acceptable salt or solvate thereof.

Second, this invention provides a method for blocking a 5HT2B receptor in a mammal, comprising administering a 5HT2B receptor occupying dose of a compound selected from the group consisting of Formula I, II, III, IV, V, VI, VII, VIII, IX, and X supra.; or a pharmaceutically acceptable salt or solvate thereof.

This invention provides a method for blocking a 5HT2B receptor in a marmnal, comprising administering a 5HT2B receptor occupying dose of a compound selected from the group consisting of Formula XI, and XII supra.; or a pharmaceutically acceptable salt or solvate thereof.

Third, this invention provides a method for selectively interacting with the 5-HT2B receptor in a mammal, comprising administering a 5-HT2B selective compound selected from the group consisting of Formula I, II, III, IV, V, VI, VII, VIII, IX, and X supra.; or a pharmaceutically acceptable salt or solvate thereof to a mammal.

This invention provides a method for selectively interacting with the 5-HT2B receptor in a mammal, comprising administering a 5-HT2B selective compound selected from the group consisting of of Formula XI and XII; or a pharmaceutically acceptable salt or solvate thereof to a mammal.

The present invention provides compounds of the Formula XI i- -L I WO 95/24200 WO 9524200PCTIUS95/03099 14 Al r~l*.<NR1R 31

I

R

3

Q'

wherein Q, is selected from the group consisting of hydrogen, R 34 and

(CHR

2

R

4 R34 is selected from the group consisting of spirobicyclic, substituted spiro-bicyclic, bicyclic or substituted bicyclic; RI is hydrogen or C 2

C

3 alkyl;

R

2 is hydrogen or Cl-C 6 alkyl; R3 is hydrogen or C 1

-C

3 alkyl;

R

4 is C5-C 8 cycloalkyl, substituted C 5

-C

8 cycloalkyl, C 5

C

8 cycloalkenyl, substituted C 5

-C

8 cycloalkenyl, bicyclic or substituted bicyclic; A is selected from the group consisting of

R

6 R7 (Ia) Ran Rs R7 (Iva) WO 95/24200 WO 9524200PCT/US95103099 where in R6 and R 7 are, independently, hydrogen, C 1

-C

6 alkyl,

C

2

-C

6 alkenyl, halo, halo (CI-C 6 alkyl, halo (C 2

-C

6 alkenyl, CORE;, C 1

-C

10 alkanoyl, C0 2

R

5

(CI-C

6 alkyl)manino, N02, or OR 5 mn is 1 or 2; is independently hydrogen or C 1

-C

4 alkyl;

R

5 is C 1

-C

4 alkyl;

R

8 is independently selected from the group consisting of an R 6 group, substituted C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl-(C-C 3 )alkyl, C 5

-C

8 cycloalkenyl, substituted C 5

-C

8 cycloalkenyl, C 5

-C

8 cycloalkenyl- (Cl-

C

3 )alkyl, C 7

-C

2 0 arylalkyl; or

R

6 and R 7 together with the carbon atoms of group A form a 5- to 8-member carbon ring;

R

3 0 and R 3 1 join t.-o form a 3 to 8 member carbon ring; or

R

3 0 and R 3 1 are independently selected from t17Th- qroup consisting of Cl-C 6 alkyl and C 2

-C

6 alkenyl; or a pharmaceutically acceptable salt or solvate the* This invention provides compounds of Formula XI

R

30 A I RR 1 X11 A is selected from the group consisting of R7 r (Ia) R8 WO 95/24200 PCT/US95/03099 16

R

6 R7 (lII a) R, and R7 (IVa) wherein

R

6 and R7 are, independently, hydrogen, Cl-C 6 alkyl,

C

2

-C

6 alkenyl, halo, halo (Cl-C6) alkyl, halo (C 2

-C

6 alkenyl,

COR

5

C

1 -Cj 0 alkanoyl, CO 2

R

5 1, (CI-C 6 alkyl) 1 marino, NO 2

-SR

5 or m is 1 or 2;

R

8 is selected from the group consisting of hydrogen, C'I-C 6 alkyl, C 2

-C

6 alkenyl, halo, halo (C 2 -C6)alkyl, halo (C 1

-C

6 )alkenyl, CaR 5

C

1

-CI

0 alkanoyl, C0 2 R5', (Cl-C 6 alkyl)mamino, NO 2

-SR

5 OR5, substituted C 3

-C

8 cycloalkyl, C 3

C

8 cycloalkyl, C 3

-C

8 cycloalkyl-(Cl-C 3 )alkyl, C5-C 8 cycloalkeny'L, substituted CS-C 8 cycloalkenyl, C 5

-C

8 cycloalkenyl- (CI-C 3 alkyl, and C 7

-C

2 0 arylalkyl; is independently hydrogen or C 1

-C

4 alkyl;

R

5 is Cl-C 4 alkyl; R6 and R 7 together with the carbon atoms of group A form a 5- to 8-member carbon ring;

R

9 and R 1 0 are independently selected from the group consisting of hydrogen, Cl-Cs alkyl, substituted C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl-(Cl-C3)alkyl, Cs-C 8 cycloalkenyl- (Cl-C 3 )alkyl, C 7

-C

2 0 arylalkyl; R11 is selected from the group consisting of CI-C 4 alkyl, OR5., fluoro, bromo, iodo, and chloro; WO 95/24200 PCT/US95/03099 17

R

3 0 and R 3 1 join to form a 3 to 8 member carbon ring; or

R

3 0 and R 3 1 are independently selected from the group consisting of CI-C6 alkyl and C 2

-C

6 alkenyl; or a pharmaceli ically acceptable salt or solvate thereof.

Finally this invention provides a method for interacting with a human 5-HT2B receptor in a human, comprising administering a 5-HT2B blocking dose of a compound selected from the group consisting of Formula I, II, III, IV, V, VI, VII, VIII, IX, and X supra.; or a pharmaceutically acceptable salt or solvate thereof to a human.

This invention provides a method for interacting with a human 5-HT2B receptor in a human, comprising administering a 5-HT2B blocking dose of a compound selected from the group consisting of Formula XI and XII; or a pharmaceutically acceptable salt or solvate thereof.

A further embodiment of this invention is an article of manufacture comprising packaging material and one or more pharmaceutical agents contained within said packaging material, wherein said pharmaceutical agent is effective for the treatment of a condition requiring 5-HT2B receptor ccupation and is selected from the group consisting of compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, and X supra.; or a pharmaceutically acceptable salt or solvate thei.of; and said packaging material comprises a label which indicates that said pharmaceutical agent can be used for the treatment of a condition requiring 5-HT2B receptor modulation.

Another embodiment of this invention is an article of manufacture comprising packaging material and one or more pharmaceutical agents contained within said packaging material, wherein said pharmaceutical agent is effective for the treatment of a condition requiring 5-HT2B receptor occupation and is selected from the group consisting of a compound of Formula XI, and XII supra.; or a pharmaceutically acceptable salt or solvate thereof; and said packaging material comprises a label which indicates that said -111~ 01 WO 95/24200 PCT/US95/03099 18 pharmaceutical agent can be used for the treatment of a condition requiring 5-HT2B receptor modulation.

Detailed Description of the Invention The term "treating" as used herein includes proph_,laxis of the named physical and/or mental condition or amelioration or elimination of the developed physical and/or mental condition once it has been established.

The terms "Ci-Cn alkyl" wherein n= 2-10, as used herein, represent a branched or linear alkyl group having from one to the specified number of carbon atoms. Typical

C

1

-C

6 alkyl groups include methyl, ethyl, n-propyl, isopropyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl and the like.

As used herein, the term "R 3 0 and R 31 join to form a 3 to 8 member carbon ring" shall mean that R 3 0 and R 3 1 are most preferably independently selected from the group consisting of Ci-C6 alkyl and C 2

-C

6 alkenyl. The carbon ring thus formed may be saturated or unsaturated. As used herein, such ring may be illustrated as: S)n; wherein n 30 shall refer to the total number of carbon atoms in the ring thus formed. Such carbon ring may be substituted with from one to four substituents independently selected from the group consisting of hydrogen, CI-Cs alkyl, NO 2 halo, halo(Ci-Cs)alkyl, halo(C2-C6)alkenyl,

C

2

-C

6 alkenyl, C0 2 R5, (Ci-C6 alkyl)mamino, -SR 5 and OR 5

A

preferred embodiment is when R 3 0 and R 3 1 join to form a C 3

-C

6 member saturated carbon ring. It is another preferred embodiment that R 3 0 and R 3 1 join to form a C3-C5 member saturated carbon ring.

When R 3 0 and R 31 do not join to form a carbon ring, it is a preferred embodiment that R 30 and R 31 are independently selected from the group consisting of C 1

-C

3 alkyl.

The terms "C2-Cn alkenyl" wherein n= 3-10, as used herein, represents an olefinically unsaturated branched or Y I- ~prrr WO 95/24200 PCT/US95/03099 19 linear group having from 2 to 10 carbon atoms and at least one double bond. The groups can be branched or straight chain. Examples of such groups include 1-propenyl, 2propenyl (-CH 2

-CH=CH

2 1,3-butadienyl

(-CH=CHCH=CH

2 1-butenyl (-CH=CHCH 2

CH

3 hexenyl, pentenyl, and the like.

The terms "halide", "halogen", and "halo" include fluorine, chlorine, bromine, and iodine. The preferred halogen is chlorine.

The terms "halo(Ci-C 6 )alkyl" and "halo(C 2

C

6 )alkenyl" refer to alkyl or alkenyl substituents having one or more independently selected halo atoms attached at one or more available carbon atoms. These terms include chloromethyl, bromoethyl, trifluoroethyl, trifluoromethyl, trifluoroethylenyl, 3-bromopropyl, 3-bromo-1-propenyl, 2bromopropyl, 2-bromo-l-propenyl, 3-chlorobutyl, 3-chloro-2butenyl, 2,3-dichlorobutyl, chloroethylenyl, 5-fluoro-3pentenyl, 3-chloro-2-bromo-5-hexenyl, 3-chloro-2-bromo-butyl, trichloromethyl, dichloroethyl, 1,4-dichlorobutyl, 3bromopentyl, 1,3-dichlorobutyl, 1,1-dichloropropyl, and the like. More preferred halo-(Ci-C6)alkyl groups are trichloromethyl, trichloroethyl, and trifluoromethyl. The most preferred halo-(Cl-C6)alkyl is trifluoromethyl.

The term "C 1

-C

1 i alkanoyl" represents a group of the formula C(O)(Ci-C 9 alkyl. Typical C 1

-CI

0 alkanoyl groups include acetyl, propanoyl, butanoyl, and the like.

The term "(Ci-C 6 alkyl)mamino" wherein m=1-2; refers to either a mono- or a dialkylamino group in which the alkyl portion of the group may be straight or branched.

Examples of such groups are methylamino, dimethylamino, ethylamino, diethylamino, 2-propylamino, 1-propylamino, di(npropyl)amino, di(iso-propyl) amino, methyl-n-propylamino, tbutylamino, and the like.

The term "C 3 -Cn cycloalkyl" wherein n=4-8, represents cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

II

WO 95/24200 PCT/US95/03099 The term "substituted(C5-Cn) cycloalkyl" refers to a cycloalkyl group as described supra wherein the cycloalkyl group may be substituted with from one to four substituents independently sele 'ed from the group consisting of hydrogen,

C

1

-C

6 alkyl, NO2, halo, halo(Ci-C6)alkyl, halo(C 2 -C6)alkenyl,

C

2

-C

6 alkenyl, CO 2 R5, (Ci-C 6 alkyl)mamino, -SR5, and The term "C 3

-C

8 cycloalkyl-(Ci-C3)alkyl" represents a linear alkyl group substituted at a terminal carbon with a

C

3 -Cs cycloalkyl group. Typical cycloalkylalkyl groups include cyclohexylethyl, cyclohexylmethyl, 3cyclopentylpropyl, and the like.

The term "C5-C 8 cycloalkenyl" represents an olefinically unsaturated ring having five to eight carbon atoms, eg., phenyl, cyclohexadienyl, cyclohexenyl, cyclopentenyl, cycloheptenyl, cyclooctenyl, cyclohexadienyl, cycloheptadienyl, cyclooctatrienyl and the like.

The term "substituted (C5-Cs) cycloalkenyl" refers to a cycloalkenyl group as described supra wherein the cycloalkenyl group may be substituted with from one to four substituent's independently selected from the group consisting of hydrogen, CI-C 6 alkyl, NO 2 halo, halo(Ci-Cg)alkyl, halo(C 2 -C6)alkenyl, C 2

-C

6 alkenyl, CORs, Ci-Cio alkanoyl, C 7

C

2 0 arylalkyl, CO 2 R5, (C 1 -Cs alkyl)mamino, -SR5, and ORs.

The term "Cs-C 8 cycloalkenyl-(C 1

-C

3 )alkyl" represents a linear CI-C 3 alkyl group substituted at a terminal carbon with a C5-C 8 cycloalkenyl gro.p.

The term "aryl" represents phenyl or naphthyl.

The aryl group can be unsubstituted or can have one or two substituents independently selected from the group consisting of Ci-Cs alkyl, C 3 -C8 cycloalkyl, substituted C 3 -Cs cycloalkyl, C 2

-C

6 alkenyl, C 3

-C

8 cycloalkyl-(C 1

-C

3 )alkyl, phenyl, Ci'-C8 cycloalkenyl, substituted C5-C8 cycloalkenyl,

C

5

-C

8 cycloalkenyl-(C1-C3)alkyl, CORs, CI-Clo alkanoyl, and C 7 -C16 arylalkyl. The substituents may be located at any available position on the .ryl ring.

The term "C 7

-C

2 0 arylalkyl" represents an aryl-(C1-

C

0 o)alkyl substituent wherein the alkyl group is linear, such i WO 95/24200 PCT/US95/03 )9 21 as benzyl, phenethyl, 3-phenylpropyl, or phenyl-t-butyl; or branched.

The term "bicyclic" represents either an unsaturated or saturated stable 7- to 12-membered bridged or fused bicyclic carbon ring. The bicyclic ring may be attached at any carbon atom which affords a stable structure.

The term includes, but is not limited to, naphthyl, dicyclohexyl, dicyclohexenyl, and the like.

The term "unsaturated bicyclic" represents a stable bicyclic ring of 7 to 12 carbon atoms. The unsaturated bicyclic ring may be attached at any carbon atom which affords a stable structure. The unsaturated bicyclic ring may be substituted with from one to four substituents as defined for "substituted bicyclic" infra.

The general term "substituted bicyclic" refers to a bicyclic ring system with up to 4 substituents attached at any desired positions on the bicyclic ring system. The bicyclic substituents may be independently selected from the group consisting of hydrogen, CI-Cs alkyl, NO 2 halo, halo(C 1

C

6 )alkyl, halo(C 2 -C6)alkenyl, C 2 -C6 alkenyl, COR 5

C

1

-C

10 alkanoyl, C 7 -C20 arylalkyl, C0 2

R

5

(CI-C

6 alkyl)mamino, and OR5; wherein R5 is defined supra. It is intended that the substituted bicyclic substituent may bond to the CHR 2 group through any available carbon atom in the bicyclic ring system. The term includes, but is not limited to compounds such as, 2-methyldicyclohexyl, 3-hydroxydicyclohexyl, benzocyclohexyl, benzocyclohexenyl, 2-methoxybenzocyclohexyl, 6-chlorobenzocyclohexenyl, 8-ethenylbenzocyclohexyl, and the like.

The term "spiro-bicyclic" and "substituted spirobicyclic" refer to a bicyclic or substituted bicyclic (as defined supra.) directly attached to the carbon of the parent ring at substituent Foi illustration purposes, a spirobicyclic is attached as shown: i~plrrrm~aar~ ill~N-; c~ WO 95/24200 PCT/US95/03099 22

R

30

R

31 Al N NR1

R

The term "naphthyl" refers to a naphthalene ring system substituent, as commonly used in organic chemi ,try.

The naphthyl substituent may bond to the CHR2 group through any available carbon atom in the naphthyl ring system. The term "substituted naphthyl" refers to a naphthyl ring system with up to 4 substituents attached at any desired positions on the naphthyl ring system. The naphthyl substituents may ba independently selected from the "substituted bicyclic" group supra.

The term "phenyl" as used herein refers to an unsubstituted benzene ring system. The term "substituted phenyl" refers to a benzene ring system with from one to thrne substituents independently selected from the group of bicyclic substituents defined supra; R5 is defined supra.

The term "CI-C 4 alkoxy" represents a straight or branched alkoxy chain having from one to four carbon atoms.

I1-C 4 alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, and the like.

In a compound of Formula IV when m' is 0, the ring attached to the amide nitrogen atom is cyclopentyl; when m is 1, the ring is cyclohexyl; when m' is 2, the ring is cycloheptyl; and when m' is 3, the ring is cyclooctyl. If the cycloalkyl ring is substituted, the substituent may be at an available position on the ring.

The term "pyridinyl" refers to or 4pyridinyl. The term "imidazolyl" refers to or 4imidazolyl.

WO 95/24200 PCT/US95/03099 23 The term "alk" refers to a divalent organic radical derived from a straight or branched Ci-C alkane. Such groups include but are not limited to -CH2-, -CH(CH 3 -C(CH3) 2

-CH(C

2

-CH

2

CH

2

-CH

2

CH(CH

3

-CH

2

C(CH

3

-CH

2

CH(CH

3

)CH

2

-CH(CH

3

)CH(CH

3

-CH(CH

3

)CH

2

CH(CH

3 and the like.

The term "optionally substituted phenyl" refers to a phenyl ring which may contain one or two substituents selected from the group consisting of C 1

-C

3 alkyl, C 1

-C

3 alkoxy, C 1

-C

3 alkylthio, halo, NO 2 and CN.

The term "selective interaction with a 5-HT2B receptor" refers to a method of interacting with the 5-HT2B receptor to a greater extent than the 5-HT2A or 5-HT2c receptor.

The term "protic acid" refers to an acid having an acidic hydrogen. Preferred protic acids include hydrochloric acid, formic acid, perchloric acid, sulfuric acid, and phosphoric acid in an aqueous medium. The most preferred protic acids are hydrochloric acid, sulfuric acid, and formic acid.

The term "organic solvent" includes solvents containing carbon, such as halogenated hydrocarbons, ether, toluene, xylene, benzene, and tetrahydrofuran.

The term "agitate" includes such techniques as stirring, centrifugation, mixing, and other similar methods.

The term "aprotic solvent" refers to polar solvents of moderately high dielectric constant which do not contain an acidic hydrogen. Examples of common aprotic solvents are dimethyl sulfoxide (DMSO), dimethylformamide, sulfolane, tetrahydrofuran, diethylether, methyl-t-butyl ether, or 1,2dimethoxyethane.

The term "protic solvent" refers to a solvent containing hydrogen that is attached to oxygen, and hence is appreciably acidic. Common protic solvents include such solvents as water, methanol, ethanol, 2-propanol, and 1butanol.

I- WO 95/24200 PCT/US95/03099 24 The term "inert atmosphere" refers to reaction conditions in which the mixture is covered with a layer of inert gas such as nitrogen or argon.

Abbreviations used herein have their accepted meaning, unless stated otherwise. For example, "Me" and "Et" refer to methyl, ethyl respectively, and "t-Bu" refers to tertiary-butyl. The abbreviation "RT" refers to room temperature or ambient conditions unless indicated otherwise.

The term "ligand" refers to compounds that are bound by the indicated receptor. Compounds useful as selective ligands may be used to selectively occupy the specific receptor site or may act as a selective agonist at the specific receptor site.

The term "substantially pure" is intended to mean at least about 90 mole percent, more preferably at least about 95 mole percent, and most preferably at least about 98 mole percent of the desired enantiomer or stereoisomer is present compared to other possible configurations.

Compounds which ara contemplated for use in modulating-a 5-HT2B receptor include, but are not limited to 7-bromo-8-methyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole, 6-isopropyl-8-methoxy-l,2,3,4-tetrahydro-9H-pyrido[3,4b]indole, 5-chloro-8-ethoxy-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole, 6-chloro-7-methyl-8-fluoro-l,2,3,4-tetrahydro-9Hpyrido[3,4b]-indole, 5-dimethylamino-8-hydroxy-l,2,3,4tetrahydro-9H-pyrido[3,4b]-indole, 6-nitro-8-butyl-l,2,3,4tetrahydro-9H-pyrido[3,4b]-indole, 7-cyclohexyl-8-hydroxy- 1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole, 6-[3-methylcyclohexyl]-8-methyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole, 6-benzyl-8-fluoro-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole, 5-cyclohexylmethyl-8-chloro-l,2,3,4-tetrahydro-9Hpyrido[3,4b]-indole, 6-carboxyl-8-bromo-l,2,3,4-tetrahydro- 9H-pyrido[3,4b]-indole, 6-ethoxy-8-isopropyl-3-methyl- 1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole, 6,8-dichloro-4naphthylmethyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole, 6,8-dimethyl-3,4-dimethyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole, 7,8-difluoro-2(N)-methyl-1,2,3,4-tetrahydro-9H- WO 95/24200 WO 95/4200 CT/US95/03099 pyrido 4b] -indole, 6, 8-dibutyl-2 -cyclopropylmethy.- 1,2 3,4-tetrahydro-9H-pyrido[3,4b1-indole, 6,8-dibromo-2 cyclohexenylniethyl-1, 2,3, 4-tetrahydro-9H-pyrido 4b1 -indole, 8-chloro-2(N) -benzyl-1,2, 3,4-tetrah-ydro-9H--pyrido 4b] indole, 8-f luoro-4-inethyl-2 -cyclohexyl-1, 2, 3,4-tetrahydro- 9H-pyrido[3,4b1-ildole, 6-iethylamine-8-chloro-1,2,3,4tetrahydro-9H-pyrido 4b] -indole, 6-chloromethyl-8-chloro- 1,2,3, 4-tetrahydro-9R--pyrido[3, 4b] -indole, 7-methoxy-1naphthylpiperazine, 1-naphthylpiperazine, 7-bromo-1H-indole- 3-ethanamine, 7-fluoro-1H--indole-3-ethanamine, 7-methoxy-lEndole-3-ethanamine, 7-chloro-1H--indole-3-ethanamile, rnethyl-7-chloro-1H-ifldole-3-ethaflamine, 1-H-Benz indole-3ethanamine, 6-methyl-7-chloro-E-indo-e-3-ethalamie, 6bromo-7-methyl-1H-indole-3-ethanaile, 6-methyl-1H-indole-3ethanamine, 5-methyl-7-bromo-1H-indole-3-ethanamine, 6,7dirnethyl-1H-indole-3-ethanamile, 6-methyl-7-bromo-1H-indole- 3-ethanamine, (81)-N-cyclohexyl-1-isopropyl-6-fl-butYlergoline-8-carboxanide, (8p)-N-cyclohexyl ethyl -1-i sopropYl 6-methylergoline-8-carboxamide, other (8j)-1--alkyl-6- (substituted)ergolines described in U.S. Patent 4,931,447, cycloalkylainides of (81)-l--alkyl-6- (substituted) ergolines described in U.S. Patent 4,981,859, compounds described in U.S. Patent 4,563,461, compounds described in U.S. Patent 4,902,691, wherein the four aforementioned U.S. Patents are herein incorporated by reference, 1,2-dimethyl-3-ethyl-5- (dimethylamino) -indole, 2- (di-n-propylamino) (isothiazol-3yl) 4-tetrahydronaphthalene, 2-ethylamino-8- (isoxazol- 3-yl) 4-tetrahydronaphthalene, 2- (N-methyl-Nbenzylamino) -8-(5-n-propyl-1,2,3-oxadiazol-4-y1V-1,2, 3 1 4 tetrahydronaphthalene, 2-diajllylamino-8- (pyrazol-3-yl) 1,2,3, 4-tetrahydronaphthalene, 2-diethylamino-8- (1,3,4oxadiazol-2-yl)-1,2,3,4-tetrahydronaphthalene, 2-(di-npropylamino)-8-(3-mfethoxypyrid2yl)l1, 2 3 4 tetrahydronaphthaa ene, 2 -benzylmethylamino-8- (3 -methoxypyrid- 2-yl) 4-tetrahydroflaphthalene, 2-benzylmethylamino-8- (benzofuran-2-yl) 4-tetrahydronaphthalene, 2dimethylamino-8- 5-triazin-2-yl) -1,2,3,4- WO 95/24200 WO 9524200PCT/US95/03099 26 tetrahydronaphthalene, 2- (di -cyclopropylmethylamino) -8- (oxazol-4-yl) 4-tetrahydronaphthalene, 2-ethylamino-8- ,,3-oxadiazol-4-yl) -thio-1, 2,3, 4-tetrahydronaphthalene, 2n-butylamino-8- (5-methoxypyrimidin-2-yl) 2,3,4tetrahydronaphthalene, 2- (di-n-propylamino) chlorooxazol-2-yl) 3,4-tetrahydronaphthaiene, 2- (di-npropylamino) (pyriinidin-2-yl) 2, 3,4tetrahydronaphthalene, 2- (di-n-propylamino) (2aiinopyrimidin-4-yl) -1,2 3, 4-tetrahydronaphthalene, 2- (di-npropylamino)-8-(3-phenyl-1,2,4-oxadiazol-5-yl)-1,2,3,4tetrahydronaphthalene, 2- (di-n-propylanino) (3-methyl- 1,2,4-oxadiazol-5-yl) -1,2,3,4-tetrahydronaphthalene, 2-(di-npropylamino) (pyrazin-2-yl) 4-tetrahydronaphthalene, 2- (di-n-propylamino) jbromopyrazin-2-yl) -1,2,3,4tetrahydronaphthalene, 2- (di-n-propylanino) (benzothiazol- 2-yl) 4-tetrahydronaphthalene., 2- (di-n-propylamino) -8- (benzoxazol-2-yl) 3,4-tetrahydronaphthalene, 2- (di-.np.ropylarnino) (indol-3-yl) 4-tetrahydronaphthalene, 3- (di-n-propylamino) (isoxazol-2-yl) -1,2,3,4tetrahydronaphthalene, 3- (di-n-propylamino) (isoxazol-2yl) -chromane, 5- (isoxazol-5-yl) (dipropylamino) chroinane, (3-met.'-ylisoxazo1-5-yl) (dipropylamino) chroinane, 5- (4- (dipropylamino)chromane, 5- (3,4- (dipropylanino) chromane, 5- (3- (dipropylamino) thiochromane, 5- (4- (dipropylamino) thiochronane, 5- (3,4- (dipropylamino) thiochronane, 8- 6, 7-tetrahydrobenz isoxazol-l-yl) -2- (direthylamino) tetrahydronaphthalene, and the like.

Especially preferred compounds for use in modulating a 5-IiT2B receptor include 7-bromo-8-methyl-l, 2,3,4tetrahydro-9H-pyrido 4b] -indole, 6-isopropyl-8-methoxy- 1,2,3, 4-tetrahiydro-9H-pyrido 4b] -indole, 5-chloro-8-ethoxy- 1,2,3, 4-tetrahydro-9H-pyrido [3,4b1 -indole, 6-cbloro-7-methyl- 8-fluoro-l,2,3,4-tetrahydro-9H-pyrido[3,4b] -indole, dimethylamino-8-hydroxy-1, 2, 3,4-tetrahydro-9H-pyridol3,4b] indole, 6-nitro-8-butyl-1,2,3,4-tetrahydro-9H-pyridol3,4b]- WO 95/24200 WO 9524200PCT/US95103099 27 indole, 7-cyclohexyl-8-hydroxy-l,2,3,4-tetrahydro-9Hp-yr.*ido[3,4b]-indole, 6-[3-methyl-cyclohexyl]-8-nethyl- 1,2,3, 4-tetrahydro-9H-pyrido[3 ,4b1 -indole, 6-benzyl-8-fluorol,2,3..4-tetrahydro-9H-pyrido[3,4b]-indole, cyclohexylmethyl-8-chloro-1, 2, 3,4-tetrahydro-9H-pyrido 4b] indole, 6-carboxyl-8-brono-l,2, 3, 4-tetrahydro-9Hpyrido[3,4b1 -indole, 6-ethoxy-8-isopropyl-1,2,3,4-tetrahydro- 9H-pyrido[3, 4b] -indole, 6, 8-dichloro-4-napahthylmethyl- 1,2,3,4-tetrahydro-9H--pyrido[3,4b]-indole, 6, 8-dimethyl-3,4dimethyl-1, 2,3, 4-tetrahydro-9H-pyrido[3, 4b1 -indole, 7,8difluoro-2 -methyl-i, 2, 3,4-tetrahydro-9H-pyrido 4b] indole, 6,8-dibutyl-2(N) -cyclopropyli~iethyl-1,2,3,4tetrahydro-9H-pyridlo[3,4b3-indole, 6,8-dibroino-2 cyclohexenyimethyl 2, 3 ,4-tetrahydro-9H-pyrido 4b] -indole, 8-chloro-2 -benzyl-l, 2,3, 4-tetrahydro-9H-pyrido 4b] indole, 8-fluoro-4-methyl-2(N)-cyclohexyl-1,2,3,4-tetrahydro- 9H-pyrido[3, 4b] -indole, 6-methylamine-8-chloro-3-isopropyll,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole, 6-chloromethyl-8chloro-l, 2,3, 4-tel- Thydro-9H-pyrido[3, 4b] -indole, 7-methoxy- I-naphthylpiperazine, 1-naphthylpiperazine, 7 -bromo-lHindole-3-ethanamine, 7-f luoro-lI--indole-3-ethanamine, 7methoxy-lH-indole-3-ethanamine, 7-chloro-1H-indole-3ethanamine, 5-methyl-7-chloro-I--indole-3-ethanamine, 1-li- Benz indole-3-ethanamine, 6-zethyl-7-chloro-lH-indole-3ethanamine, 6-bromo-7-methyl-1H-indole-3-ethanamine, 6iethyl-1H-indole-3-ethanamine, 5-methyl-7-bromo-1H-indole-3ethanamine, 6, 7-dimethyl-lH-indole-3-ethanamine, 6-rnethyl-7bromo-1H-indole-3-ethanamine, 1, 2-dirnethyl-3-ethyl-5- (diiethyl amino) indol1e, 2- (di-n-propylamino) (isothiazol-3yl) 2,3, 4-tetrahydronaphthalene, 2-ethylamino-8- (isoxazol- 3-yl)-l,2,3,4-tetrahydronaphthalene, 2- (N-methyl-Nbenzylamino)-8-(5-n-propyl-1,2,3-oxadiazol-4-yl)-1,2,3,4tetrahydronaphthalene, 2-diallylamino-8- (pyrazol-3-yl) 1,2,3,4-tetrahydronaphthalene, 2-diethylaai.no-8-(1,3,4oxadiazol-2-yl) -1,2,3,4-tetrahydronaphthalene, 2-(di-npropylamino) (3-methoxypyrid-2-yl) -1,2,3,4tetrahydronaphthalene, 2 -benzylmethyl amino -8 (3 -methoxypyrid- WO 95/24200 WO 95/4200 CTIUS95/03099 28 2-yl) 2,3, 4-tetrahydronaphthalene, 2-benzylmethylanino-8- (benzofuran-2-yl) 4-tetrahydronaphthalene, 2dimethylamino-8-C1,3,5-triazin-2-yl)-1,2,3,4tetrahydronaphthalene, 2- (di-cyclopropyrnethylanino) -8- (oxazol-4-yl) 2,3, 4-tetrahydronaphthaleie, 2-ethylainino-8- (1,2,3-oxadiazol-4-yl)-thio-1,2,3,4-tetrahydronaphthalene, 2ri-butylanino-8-(3-methoxypyrimidin-2-yl) -1,2,3,4tetrahydronaplithalene, 2- (di-n-propylamino) chlorooxazol-2-yl) 4-tetrahydronaphthalene, 2- (di-npropylamino) (pyrirnidin-2-yl) -1,2,3,4tetrahydronaphthalene, 2- (di-n-propylamino) (2aminopyrimidin-4-yl) 2,3, 4-tetrahydronaphthalene, 2- (di-npropylamino)-8-(3-phenyl-1,2,4-oxadiazol-5-yl)-1,2,3,4tetrahydronaphthalene, 2- (di-n-propylanino) (3-methyl- 1,2,4-oxadiazol-5-y1) -1,2,3,4-tetrahydronaphthalene, 2-(di-npropylamino) (pyrazin-2-yl) 4-tetrahydronaphthalene, 2- (di-n-propylamino) (bromopyrazin-2-yl) 2,3,4tetrahydronaphthalene, 2- (di-n-propylanino) (benzothiazol- 2-yl) 2,3, 4-tetrahydronaphthalene, 2- (di-n-propylamino) -8- (benzoxazol-2-yl)-1,2,3,4-tetrahydronaphthalene, 2-(di-npropylamino) (indol-3-yl) 4-tetrahydronaphthalene, (isoxazol-5-yl)-3-(dipropylanino)chronane, 5-(3- (dipropylarnino) chroinane, 5- (4- (dipropylainino) chroniane, 5- (3,4- (dipropylamino) chroinane, 5- (3- (dipropylamino) thiochronane, 5- (4- (dipropylamino) thiochromane, 5- (3,4- (dipropylainino) thiochronane, 8- 6, 7-tetrahydrobenz [ci isoxazol-1-yl) -2- (diinethylainino) tetrahydronaphthalene, and 3- (di-npropylanino) (isoxazol-2-yl) 3, 4-tetrahydronaphthalene.

A preferred compound of Formula IX has the following structure: WO 95/24200 PCT/US95/03099 29

R

28

N

O R 29

I

S7R 2 N. R27 ya Wherein R 2 6

R

2 7

R

2 8

R

2 9 and ya are as defined supra.

When Q is hydrogen, preferred compounds of Formula I have the following structure: R 7

R

8 NNR1

R

6

H

Wherein R 6 is selected from the group consisting of Ci-C 4 alkyl, OR5,, fluoro, bromo, and chloro; is C 1

-C

4 alkyl; and RI, R 7 and R8 are as defined supra.

The 5-HT2B receptor has been identified in various tissues and organs in the rat. The primary areas of 5-HT2B receptor localization in the rat include lung, uterus, bladder, stomach, and colon. Further, the 5-HT2B receptor has been identified in various tissues and organs in the human.

Interesting areas of 5-HT2B receptor loca"ization in the human include but are not limited to the brain and blood vessels.

In light of the receptor localization, physiological conditions which can be mediated by the 5-HT2B receptor include incontinence, bladder dysfunction, Functional Bowel Disorders, stomach emptying disorders, respiratory disorders including asthma, uterine dysfunction including endometriosis, fibrosis, and motility disorders such as but not limited to induction of labor, sleeping ~llii" IY__ WO 95/24200 PCT/US95/0309 disorders, eating disorders, including bulimia and obesity, consumption disorders, thermoregulation, sexual disorders, hyperactivity, excessive aggression, alcoholism, anxiety, obsessive-compulsive disorders, depression, psychosis, schizophrenia and schizophreniform disorders, panic disorders, Gilles de la Tourette syndrome, and Alzheimer's disease and cardiovascular diseases such as thrombosis, hypertension, vasospasm (peripheral and/or central) such as stroke, angina, and other vascular occlusive diseases.

Further, migraine headaches can be treated using 5-HT2B receptor stimulating compounds of this invention. Preferred examples of such conditions which may be treated using 5-HT2B modulators include cardiovascular disorders, uterine dysfunction, sleep disorders, hallucinogenic activity, psychosis, anxiety, depression, thermoregulation, feeding disorders, and hypotension. See Leonard, International Clinical Psvchopharmacoloqv, 7, 13-21 (1992). It is particularly preferred to use a 5-HT2B antagonist for treating a Functional Bowel Disorder.

Several examples of more specific CNS disorders which may be treated using 5-HT2B modulating compounds of this invention include, but are not limited to: (numerals in parenthesis refer to the DSM-III-R Classification Codes) Attention-deficit hyperactivity disorder (314.01), conduct disorders (312.20, 312.00, 312.90), primary degenerative dementia of the Alzheimer type, senile onset (290.30, 290.20, 290.21, 290.00), primary degenerative dementia of the Alzheimer type, presenile onset (290.11, 290.12, 290.13, 290.10), alcohol withdrawal delirium (291.00), alcohol hallucinosis (291.30), alcohol, dementia associated with alcoholism (291.20), cannabis, delusional disorder (292.11), cocaine, intoxication (305.60), hallucinogen, mood disorder (292.84), nicotine withdrawal (292.00), phencyclidine or similarly acting arylcyclohexylamine intoxication (305.90), other psychoactive substance intoxication (305.90), delirium (293.00), dementia (294.10), organic delusional disorder (293.81), organic hallucinosis (293.82), organic mood i r i MEN~ WO 95/24200 PCTUS95/03099 31 disorder (293.83), organic anxiety disorder (294,80), organic personality disorder (310.10), organic mental disorder (294.80), schizophrenia, catatonic (295.21, 295.22, 295.23, 295.24, 295.25, 295.20), schizophrenia, disorganized (295.11, 295.12, 295.13, 295.14, 295.15, 215.00), schizophrenia, paranoid (295.31, 295.32, 295.33, 295.34, 295.35, 295.00), schizophrenia, undiffertiated (295.91, 295.92, 295.93, 295.94, 295.9E, 295.00), schizophrenia, residual (295.61, 295.62, 295.63, 295.64, 295.65, 295.60), delusional (paranoid disorder (297.10), schizophreniform disorder (295.40), schizoaffective disorder (295.70), induced psychotic disorder (297.30), bipolar disorder, mixed (296.61, 296.62, 296.63, 296.64, 296.65, 296.66, 296.60), bipolar disorder, manic (296.41, 296.42, 296.43, 296.44, 296.45, 296.46, 296.40), bipolar disorder, depressed (296.51, 296.52, 296.53, 296.54, 296.55, 296.56, 296.50), major depression, single episode (296.21, 296.22, 296.23, 296.24, 296.25, 296.26, 296.20), major depression, recurrent (296.31, 296.32, 296.33, 296.34, 296.35, 296.36, 296.30), obsessive compulsive disorder (300.30), post-traumatic stress disorder (309.89), generalized anxiety disorder (300.02), hypochondriasis (300.07), somatization disorder (300.81), male erectile disorder (302.72), intermittent explosive disorder (312.34), impulse control disorder (312.39), paranoid (301.00), schizoid (301.20), schizotypal (301.22), antisocial (301.70), and borderline (301.83). Diagnostic and Statistical Manual of Mental Disorders, 3rd Ed. Revised, (1980), prepared by the Task Force on Nomenclature and Statistics of the American Psychiatric Association.

Thus, the present invention also provides methods for treating or preventing the above-named conditions.

The skilled artisan will recognize that psychosis or psychotic conditions are characterized by hallucinations, delusions, or grossly disorganized behavior which indicate that the patient suffers from gross impairment in reality testing. Therefore, drugs having antipsychotic activity can I L I. ~-p WO 95/24200 PCT/US95/03099 32 be useful for treating a variety of important psychotic conditions.

As used herein the term "Functional Bowel Disorder" refers to a functional gastrointestinal disorer manifested by abdominal pain and/or symptoms of disturbed defecation (urgency, straining, feeling of incomplete evacuation, altered stool form [consistency] and altered bowel frequency/timing) and/or bloating (distention).

The term "Functional Bowel Disorder" includes but is not limited to irritable bowel syndrome, hypermotility, ichlasia, hypertonic lower esophogeal sphinctor, tachygastria, constipation, hypermotility associated with irritable bowel syndrome.

Functional Bowel Disorders are characterized by abnormal bowel function without detectable structural abnormalities. Abnormal bowel function includes diarrhea, constipation, mucorrhea, and pain or discomfort over the course of the sigmoid colon. Such disorders are influenced by psychological factors and stressful life situations.

'The Functional Bowel Disorder, Irritable Bowel Syndrome (IBS), is one of the most commonly encountered gastrointestinal disorders. Between 20% and 50% of patients referred to gastrointestinal clinics suffer from IBS.

Symptoms of IBS occur in approximately 14% of otherwise apparently healthy people. IBS is a complex condition, in part because it is not a disease but a syndrome composed of a number of conditions with similar manifestations.

Current therapy for Functional Bowel Disorders is restricted to drugs which treat only a small proportion of patients. For example, anticholinergic drugs reduce spasticity, therefy relieving some of the abdominal pain.

Histamine H2 receptor antagonists inhibit gastric acid secretion and my relieve some dyspeptic symptoms. A therapeutic agent that relieves most of the Functional Bowel Disorder symptoms is currently not available.

The term Functional Bowel Disorder includes conditions such as Irritable Bowel Syndrome, ichlasia, WO 95/24200 PCTUS95/03099 33 hypertonic lower escphogeal sphincter, tachygastria, hypermotility associated with irritable bowel syndrome, and constipation.

The compounds described herein can form acid addition salts with a wide variety of inorganic and organic acids. Typical acids which can be used include sulfuric, hydrochloric, hydrobromic, phosphoric, hypophosphoric, hydroiodic, sulfamic, citric, acetic, maleic, malic, succinic, tartaric, cinnamic, benzoic, ascorbic, mandelic, ptoluenesulfonic, benzenesulfonic, methanesulfonic, trifluoroacetic, hippuric and the like. The pharmaceutically acceptable acid addition salts are especially preferred for the treatment of 5-HT2B receptor related conditions.

Certain compounds are preferred for use in treating conditions related to the modulation of a 5-HT2B receptor.

The following invention embodiments and compound characteristics listed in tabular form may be -ndependently combined to produce a variety of preferred compounds and embodiments of the invention. The following list of embodiments of this invention is in no way intended to limit the scope of this invention in any way: A) R 1 is hydrogen; B) R 2 is hydrogen or methyl; C) R 3 is hydrogen or methyl; D) R 4 is CS-C 8 cycloalkenyl or substituted C 5 -Cs cycloalkenyl, bicyclic or substituted bicyclic, wherein the substituents are selected from the group -onsisting of hydrogen, C 1

-C

6 alkyl, NO 2 halo, halo(C 1

-C

6 )alkyl, C 2

-C

6 alkenyl, COR 5

(CI-C

6 alkyl)mamino, -SR 5 and OR 5 E) A is a group of formula III; F) A is a group of formula IV wherein R 6 and R 7 are CI-C 6 alkyl or halo, and R 8 is hydrogen, C 1 -CS alkyl, halo, C 5

-C

8 cycloalkyl, phenyl or substituted-phenyl; G) The compound interacting with the 5-HT2B receptor is a 5-HT2B receptor antagonist; H) The compound interacting witht he 5-HT2B receptor is a 5-HT2B receptor partial agonist, WO 95/24200 PCT/US95/03099 34 I) R4 is substituted C5-C 8 cycloalkenyl; wherein the substituents are selected from the group consisting of hydrogen, NO 2 halo, (Ci-C6 alkyl)mamino, and J) A is a group of formula IV wherein R 6 is hydrogen, R7 and RS are independently selected from the group consisting of halo and Ci-C 4 alkyl; K) R 4 is naphthyl or substituted naphthyl wherein the naphthyl substituents are selected from the group consisting of (CI-C 6 alkyl)mamino and ORs; L) ya is CH 2

R

2 6 and R 2 7 are each C 2

-C

3 alkyl; and R 2 8 and R 29 are each hydrogen; M) Compounds of the Formula I, II, III, IV, and V- N) Compounds of the Formula II, III, and VIII; O) Compounds of the Formula VI, VIII, IX, XI, and XII; P) A compound of the Formula X; Q) Compounds of wherein R6 is methyl, R 2 is methyl, and R4 is substituted alkenyl wherein the alkenyl group is phenyl and there are two substituents which are each methoxy; R) The 5-HT2B modulated condtion is a Functional Bowel Disorder.

S) The Functional Bowel Disorder is irritable bowel syndrome.

T) The 5-HT2B modulated condition is psychosis.

U) The 5-HT2B selective compound has a greater affinity for 5-HT2B receptors than it has for 5 -HT2A receptors.

V) The 5-HT2B selective compound has a greater affinity for 5-HT2B receptors than it has for 5-HT2c receptors.

W) The 5-HT2B modulated condtion is selected from the group consisting of urinary incontinence, bladder dysfunction, uterine dysfunction, cardiovascular disorder, and respiratory disorder.

X) The compound is administered in a unit dosage form.

Y) The label on the article of manufacture states that WO 95/24200 PCT/US95/03099 the compound is useful for treating a condition selected from the group consisting of urinary incontinence, bladder dysfunction, uterine dysfunction, cardiovascular disorder, respiratory disorder, and Functional Bowel Disorder.

Z) A pharmaceutical formulation comprising one or more pharmaceutically acceptable exipients and a 5-HT2B receptor modulating compound; Zl) A compound wherein R 4 is aromatic; Z2) A compound wherein R 4 is an aromatic bicyclic; Z3) A compound of Formula V7I.

Certain compounds of Formula II are useful for modulating 5HT2B receptors. Ce .in compounds of Formula II within the scope of this invention are preferred for that use. The following invention embodiments and compound characteristics listed in tabular form may be independently combined to produce a variety of preferred compounds and embodiments of the invention. The following list of embodiments of this invention is in no way intended to limit the scope of this invention in any way.

A) R 9 and RIo are each hydrogen.

B) R 11 is C 1

-C

3 alkyl.

C) R11 is chloro, fluoro, or bromo.

D) R 11 is -OCH 3 E) R 6 is C 1

-C

4 alkyl.

F) R6 is methyl.

G) A method for binding a 5HT2B receptor using one or Scompounds of Formula I and/or II.

H) A method of using one or more compounds of Formula I and/or LI for trpting a functional bowel disorder.

I) A method of using one or more compounds of Formula I and/or II which are useful for stimulation of the 5HT2B receptor for treating a condition selected from the group consisting of urinary incontinence, bladder dysfunction, uterine dysfunction, cardiovascular disorders, and respiratory disorders.

J) A method for using one or more compounds of Formula I and/or II for treating Irritable Bowel Syndrome.

8 blu~ Y-urrsmr~r~-~~o*Ole;lse~ III L~CIC WO 95/24200 PCT/US95/03099 36 K) A pharmaceutical formulation comp.-ising a compound of Formula I or II and one or more pharmaceutically acceptable excipients.

The compounds of the present invention are useful for modulating or blocking the 5-HT 2 receptor. Certain of the present compounds of Formula XI and XII are preferred for that use. The following invention embodiments and compound characteristics listed in tabular form may be independently selected or combined to produce a variety of preferred compounds and embodiments of the invention. The following list of embodiments of this invention is in no way intended to limit the scope of this invention in any way.

A) R 1 is hydrogen; B) R 2 is hydrogen or methyl; C) R3 is hydrogen or methyl; D) R 4 is C 5

-C

8 cycloalkenyl or substituted Cs-C8 cycloalkenyl, wherein the substituents are selected from the group consisting of hydrogen, Ci-C 6 alkyl, N 2 halo, halo(Ci-

C

6 )alkyl, C 2

-C

6 alkenyl, CJR 5 (CI-Cg alkyl)m amino, -SR 5 and E) A is a group of formula III; F) A is a group of formula IV wherein R6 and R 7 are CI-C 6 alkyl or halo, and R 8 is hydrogen, C 1 -Cs alkyl, halo, Cs-C 8 cycloalkyl, phenyl or substituted-phenyl; G) R 2 is hydrogen; H) R 3 is hydrogen; I) R4 is substituted C 5

-C

8 cycloalkenyl; wherein the substituents are selected from the group consisting of hydrogen, NO 2 halo, (C 1

-C

6 alkyl)m amino, and ORs; J) A is a group of formula IV wherein R 6 is hydrogen, R7 and R8 are independently selected from the group consisting of halo and CI-C4 alkyl.

K) Q' is (CHR 2

)R

4 L) R 3 0 and R 3 1 join to form a 3 to 6 member carbon ring; M) R 3 0 and Rp join to form a 3 to 5 member carbon ring; N) R 30 and R 31 are each methyl; 0) R 4 is naphthyl; WO 95/24200 PCT/US95/03099 37 P) R, -s an optionally substituted bicyclic hydrocarbon ring system having 7 to 12 carbon atoms and 0, 1, 2, or double bonds; Q) R4 is a 6 to 10 carbon atom unsaturated bicyciic ring system; R) Q' is bicyclic or substituted bicyclic; S) R 3 4 is Ra T) R34 is an optionally substituted bicyclic ring substituent; U) R9 and Rio are each hydrogen; V) R9 is selected from the group consisting of CI-C6 alkyl, substituted C 3

-C

8 cycloalkyl, C 3

-C

8 cycloalkyl, C 3 -Ca cycloalkyl-(C 1 -C3)alkyl, C5-C 8 cycloalkenyl-(Ci-C 3 )alkyl, C7-

C

2 0 arylalkyl; W) R4 is aromatic; X) R34 is spiro-bicyclic or substituted spirobicyclic; Y) Q' is hydrogen.

The more preferred classes have the following features: A-C, E or F, I, L, N, P, R, and W.

The most preferred class of compounds has the following features: A, G-J, M, and Q.

The preferred classes of compounds for use as selective 5-HT2B ligands have the following features: A-D, E or J, M, and 0.

The most preferred class of compounds for use as selective 5-HT2B ligands has the following features: A, G-J, M, and 0.

Compounds of Formulas XI and XII are particularly useful for modulating 5HT2B receptors. Certain compounds I~-~ss WO 95/24200 PCT/US95/03099 38 within the scope of this invention are preferred for that use. The following invention embodiments and compound characteristics listed in tabular form may be independently selected or combined to produce a variety of preferred compounds and embodiments of the invention. The following list of embodiments of this invention is in no way intended to limit the scope or this invention in any way.

A) R 9 and Rio are each hydrogen; B) R11 is C 1

-C

3 alkyl; C) R 11 is chloro, fluoro, or bromo; D) R 11 is -OCH 3 E) R 30 and R 31 join to form a 3 to 8 member carbon ring; F) R 3 0 and R 3 1 join to form a 3 to 6 member carbon ring; G) A compound having preferred characteristics described supra.; H) A method for binding a 5HT2B receptor using one or more compounds of Formula XI and/or XII; I) A method of using one or more compounds of Formula XI and/or XII for treating a functional bowel disorder.

I) A'method of using one or more compounds of Formula XI and/or XII which are useful for modulatation of the 5HT2B receptor for treating a function bowel disorder.

J) A method for using one or more compounds of Formula XI and/or XII for treating Irritable Bowel Syndrome.

K) A pharmaceutical formulation comprising a compound of Formula XI and/or XII and one or more pharmaceutically acceptable excipients.

Examples of compounds of Formula XI include but are not limited to: 10-methyl-2,3,4,4a,5,6,7,llc-octahydro-lH-indolo[2,3c]quinoline, 8-chloro-2,3,4,4a,5,6,7,llc-octahydro-lHindolo[2,3-c]quinoline, 6-(2,4-dimethoxybenzyl)-10-methyl- 2,3,4,4a,5,6,7,llc-octahydro-lH-indolo[2,3-c]quinoline, 7fluoro-6-(2,4-dimethoxybenzyl)-10-methyl-2,3,4,4a,5,6,7,llcoctahydro-lH-indolo[2,3-c]quinoline, 8-methoxy-6-(2,4dimethoxybenzyl) -0-methyl-2,3,4,4a,5,6,7, llc-octahydro-lHindolo [2,3-c]quinoline, 7-nitro-6-(3,4-dimethoxybenzyl)-10c WO 95/24200 WO 9524200PCTIUS95/03099 39 methyl-2,3,4,4a, 5, 6,7, llc-octahydro-lH-indolo [2,3ci quinoline, 5- 4-dimethoxybenzyl) 2,3,4,4a,5, 6,7,llc-octahydro-1H-indolo[2,3-c]quainoline, 7- 4-diznethoxybenzyl) -l0-methyl-2, 3,4, 4a, 5, 6,7,ll~coctahydro-lH-indolo[2,3-clquinoline, 6-ethoxy-5-Q' 4dimethoxybenzyl4) -lO-methyl-2, 3, 4, 4a, 5, 6, 7, llc-octahydro-1Hindolo[12, 3-clquinoline, 7 -nitro- 6- 4-dimethoxybenzyl) -10 methyl-2,3,4,4a,5,6,7,10c-octahydro-lH-indolo[2,3ci quinoline, 7- 4-dimethoxybenzyl) 2,3,4,4a,5,6,7,llc-octahydro-1H-indolo[2,3-c]quinol.ine, 7nitro-6-(3,4-diethoxybenzyl)-10-methyl-2,3,4,4a,5,6,7,llcoctahydro-1H-indolo 3-c~quinoline, 6-methyl-8-bromo-1- (3,4-dimethoxyphenyl)-l0-inethyl-2,3,4,4a,5, 6,7,11coctahydro-lH-indolo[12, 3-cIquinoline, 7- 1-dimethylethyl) (I1-naphthalenyl -1-ethyl) 3, 4,4a, 5, 6, 10 c-pyrido [3,4-b] indole hydrochloride, 7-methyloxy-1- (2methyilaminonaphthalenyl) -1-ethyl)-1,2,3,4,4a,5,6,10coctahydrocyclopenta[alpyrido 4-b] indole, 2butenedioate, 6- (l,1-dimethylethyl)-1-(1-(3diethylam'lnonaphthalenyl) -1-ethyl) 4a, 5,6, l0coctahydrocyclopenta [alpyrido- 13, 4-ba indole hydrochloride, and 11(4-dimethylamino-naphthalenyl) -methyl] 1,2,3,4, 4a, 5,6, 10c-octahydrocyclopenta[a]pyrido- 4-blindole dihydrochioride.

Examples of compounds of Formula XII include but are not limited to: 3- (2-amine-cyclopentyl) 7-dimethylindole, 3- (2-aminecyclopentyl) -5-methyl-7-bromoindole, 3- (2-ainine-cyclopentyl) 6-methyl-7-chloroindole, 3- (2-amine-cyclopentyl) -6-bromo-7methylindole, 3- (2-amine-cyclopentyl) -Benz indole, 3- (2amine-cyclohexyl) -5-methyl-7-chloroindole, 3- (2-aminecyclohexyl) -7-chloroindole, 3- (2-amine-cyclopropyl) -7methoxyindole, 3- (2-amine-cycloheptyl) -7-f luoroindole, 3- (2amine-cyclohexyl) -7-bromoindole, 3- (2-amine-cyclopropyl) -6methyl-7-bromoindole, 3- (2-amine-cyclopentyl) -5-f luoro-7methoxyindole, 3- (2-amine-cyclopentyl) -5-nitro-7- WO 95/24200 PCT/US95/03099 chloroindole, 3-(2-amine-cyclooctyl)-2-ethyl-7-fluoroindole, and 3-(2-amine-cycloheptyl)-2-methyl-7-fluoroindole.

The compounds which are useful for blocking 5-HT2B receptors contemplates racemic mixtures as well as the substantially pure stereoisomers of the compounds of Formulas I through XII. The term "enantiomer" is used herein as commonly used in organic chemistry to denote a compound which rotates the plane of polarization. Thus, the enantiomer" rotates the plane of polarized light to the left, and contemplates the levorotary compound of Formulas I through XII. The and enantiomers can be isolated using wellknown classical resolution techniques. One particularly useful reference which describes such methods is JACQUES et.

al. ENANTIOMERS, RACEMATES, AND RESOLUTIONS (John Wiley and Sons 1981). Appropriate resolution methods include direct crystallization, entrainment, and crystallization by optically active solvents. Chrisey, L.A. Heterocycles, 267, (1990). A preferred resolution method is crystallization with an optically active acid or by chiral synthesis as described in Example 46 using the method of A.I. Meyers.

Loewe, M.F. et al., Tetrahedron Letters, 3291, 26 (1985), Meyers, A.I. et al., J. Am. Chem. Soc., 4778, 110 (1988).

Preferred optically active acids include camphorsulfonic and derivatives of tartaric acid.

The present invention encompasses both the R and the S configurations. The terms and are used herein as commonly used in organic chemistry to denote the specific configuration of a chiral center. See, R.T. Morrison and R.N. Boyd, Oraanic Chemistry, pp 138-139 (4th Ed. Allyn Bacon, Inc., Boston) and Orchin, et al. The Vocabulary of Organic Chemistry, p. 126, (John Wiley and Sons, Inc.).

For example, the present invention includes, but is not limited to, the use of compounds such as methyl-8-bromo-l- [(3,4-dimethoxyphenyl)methyl]-1,2,3,4tetrahydro-9H-pyrido[3,4-b]indole; (-)-(S)-5,7-dimethyl- 1,2,3,4-tetrahydro-l-[(3,4-dimethoxyphenyl)methyl] -9H- L ~pe~ WO 95174200 WO 95/A200 CT/US95/03099 41 pyridoll3,4-bllindole; (-)-(S)-5-fluoro-6-methyl-l-[(2-chloro- 3, 4-dimethoxyphenyl)methyl] 4-tetrahydro-9H-pyrido [3,4blindole; and (-)-(S)-6-methyl-1,2,3,4-tetrahvdro-l-[(3,4dimethylphenyl) methyl] -9H-pyrido indole. The invention also includes, but is not limited to, the use of methyl-8-bromo-1-[ (3,4-dimethoxyphenyl)methylll-l,2,3,4tetrahydro-9H--pyridoll3,4-blindole; (+)-(S)-5,7-dimethyl- 1, 2,3, 4-tetrahydro-l-[1(3, 4-dimethoxyphenyl)methyl] -911pyrido[3,4-blindole; (+)-(S)-5-fluoro-6-methyl-1-[(2-chloro- 3 ,4-dimethoxyphenyl)methyl] 2,3, 4-tetrahydro-9H-pyrido [3,4blindole; (-)-(R)-7-methyl-8-bromo-1-[(3,4dimethoxyphenyl)methyl] 4-tetrahydro-911-pyrido [3,4blindole; (-)-(R)-5,7-dimethyl-1,2,3,4-tetrahydro-l-[ (3,4dimethoxyphenyl)methyll-9H-pyrido[3,4-b]indole; fluoro-6-methyl-l- 11(2-chloro-3, 4-dimethoxyphenyl)methyl] 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole; and methyl-i, 2,3, 4-tetrahydro-l- 4-dimethylphenyl)methyl] -911pyrido[3,4-blindole; (+)-(R)-7-methyl-8-bromo-l-[(3,4dimethoxyphenyl)methyl] 4-tetrahydro-9H--pyrido [3,4blindole; (+)-(R)-5,7-dimethyl-1,2,3,4-tetrahydro-l-[ (3,4dimethoxyphenyl)methyl]-9H-pyrido[3,4-b]indole; fluoro-6-methyl-l- 11(2-chloro-3 ,4-dimethoxyphenyl)methyl] l,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole; and methyl-i, 2,3, 4-tetrahydro-l- 4-dimethylphenyl)methyl] -911pyrido 4-b] indole.

The compounds which are useful. for interaction with 5-HT2B receptors are known to form hydrates and solvates with appropriate solvents. Preferred solvents for the preparation of solvate forms include water, alcohols, tetrahydrofuran, DMF, and DUSO. Preferred alcohols are methanol and ethanol.

Other appropriate solvents may be selected based on the size of the solvent molecule. Small solvent molecules are preferred to facilitate the corresponding solvate formation.

The solvate or hydrate is typically formed in the course of recrystallization or in the course of salt formation. One useful reference concerning solvates is Sykes, Peter,A Guidebook to-.Mechanism in Organic Chemistry, 6, 56, (1986, WO 95/24200 PCT/US95/03099 42 John Wiley Sons, New York). The term "solvate" as used herein includes hydrate forms such as monohydrate and dihydrates.

Some of the compounds which are useful for interaction with 5-HT2B receptors are either known in the art or readily available by routine synthetic processes. For example, compounds of Formula III can be prepared using the methods taught in Semonsky et al., J.K. Patent No. 816,273 (July 8, 1959), U.S. Patent Nos. 2,736,728 and 2,774,763 which U.S. Patents are hereby incorporated by reference.

Compounds of Formula IV may be prepared as described in U.S.

Patent Nos. 4,981,859 and 4,931,447 which are hereby incorporated by reference. A process for preparing compounds of Formula V is described in U.S. Patent No. 4,902,691 which is hereby incorporated by reference. A process for preparing compounds of Formula VI is described in U.S. Patent No.

4,563,461 which is hereby incorporated by reference. A compound of Formula VII can be prepared as described in Forbes, J. Med. Chem., 36:1104-1107 (1993). A compound of Formula VIII is known in the art and may be purchased or prepared by recognized methods. A process for preparing compounds of Formula IX is available to the artisan in the published European Patent application. The European Publication number is 0498590 Al (August 12, 1992; Bulliten 92/33) and is readily available to the United States artisan in the English language. A compound of Formula X is known in the art and can be prepared by recognized methods.

The compounds of the present invention can be prepared using chemical processes that are understood in the art; however, the most preferred method for preparing the formula I compounds of this invention utilizes the process of Scheme V. The most preferred method for preparing a compound of Formula 1I is using the general method illustrated in Scheme II infra. Compounds of Formula II wherein R 9

R

12 and/or R 10 are not hydrogen can be prepared using accepted chemical methods such as reductive alkylation and direct alkylation of the corresponding tryptamine.

WO 95/24200 PCT/US9503099 43 A compound of Formula I, wherein Q is hydrogen, may be prepared by contacting glyoxylic compound of formula with an amine of formula This Pictet-Spengler type reaction is generally applicable, provides desirable yields, and produces stable intermediates. Further, the product of the reaction typically may be directly isolated as the desired salt.

The compounds of formula which may be used as starting materials for the compounds of the instant invention can be purchased from arc-recognized vendors or may be prepared using well-known chemical techniques. The compounds of formula which are useful as starting materials for the compounds of this invention may be prepared as represented by Scheme I. The R 4 group is as defined herein above.

The process for preparing the compounds of this invention will be discussed in greater detail in the following paragraphs.

I--

WO 95/24200 PCT/US95/03099 44 Scheme I o R 4 o-H a CO 2

H

a N Me b Compound in Scheme I may be substituted or unsubstituted depending on the desired product. Most formula compounds necessary for the preparation of the azalactone starting materials are commercially available.

Additional substituted formula compounds may prepared using common chemical methods. Furniss, B.S. et al., Vocel's Textbook of Practical Organic Chemistry (John Wiley, New York, N.Y. 1989) see especially pp 989 through 993.

Generally, the Scheme I reaction is begun by preparing a solution of compound acetylglycine and sodium acetate in acetic anhydride. The reaction is commonly heated from about 90 0 C to about 1100C for a period of about 2hours. 'The reaction mixture is cooled to about ambient temperature and stirred for a period of about 0-10 hours under inert conditions. The reaction time will vary depending on the degree of substitution on the R 4 group and the completion of reaction desired.

When the reaction is complete, the mixture is poured onto ice with stirring. The azalactone may be isolated by standard isolation techniques such as filtration and may be dried under reduced pressure.

Compound in Scheme II is used as a starting material for compounds of Formula I. These compounds are commercially available or may be prepared using the wellknown Fischer indole synthesis applied to tryptamines. The Fischer synthesis is represented by Scheme II. is as hereinabove defined.

WO 95/24200 PCT/US95/03099 Scheme II

NNH

2 l 2 N d 0

H

C1 ~H The chlorobutanal compound used in Scheme II may be prepared through the hydrogenation of chlorobutyryl chloride. The hydrogenation may be facilitated by the use of a catalyst such as Pd/C. Other halobutanal compounds may be suitable for the Scheme II process. The starting compounds in Scheme II may be purchased or prepared using known methods. March, Advanced Oraanic Chemistry Reactions, Mechanisms, and Structure, 3rd (John Wiley Sons, New York, 1985) see especially page 1163.

The Fischer synthesis is commonly begun by adding a suitable saturated base like sodium carbonate to a stirred suspension of the hydrazine salt in an organic solvent like chloroform. The hydrazine hydrochloride salt is one especially preferred hydrazine salt. The desired hydrazine free base is extracted with the organic phase. The oil is placed in an alcohol and water solution and treated with an appropriate base like sodium acetate. The halobutanal is added and the tube purged with an inert gas like nitrogen.

The resulting mixture is placed in an oil bath which has been heated to about 90 0 C-110 0 C. The mixture should be heated for about 17 to 19 hours. The mixture is allowed to cool to ambient temperature and is concentrated under reduced pressure. The residue is partitioned between a suitable organic and basic aqueous phase, such as chloroform/methanol and aqueous sodium carbonate. The organic phase may be concentrated and the resulting compound purified by standard methods such as flash chromatography. If chromatography is used, fractions containing product may be

U-

WO 95/24200 PCTUS95/03099 46 combined and concentrated. The oil is dissolved in an appropriate solvent, such as diethyl ether containing about 1% alcohol. A preferred alcohol is methanol. The mixture may be treated with dry acid gas, such as dry HC1 gas to produce the corresponding acid addition salt of the desired compound One method for preparing Formula I compounds uses the Pictet-Spengler reaction as represented by Scheme III.

The substituents are as defined hereinabove.

I' WO 95/24200 PCT/US95/03099 47 Scheme III l NH2 R2 sCHO Al N A NH

R

3 RR4

R

3 R 4

R

2 (e) (f) Generally, the Scheme III reaction is carried out by reacting compound with the selected aldehyde in a suitable solvent such as ethanol or methanol for a period of about 1 to 50 hours depending on the desired product. The reaction may be refluxed if necessary. The precipitated reaction product is collected by common isolation methods, such as filtration and may be purified by recrystallization.

If a compound with an R 1 substituent is desired, the reaction may be followed by a reductive alkylation. The reductive alkylation is represented by Scheme IV.

Scheme IV N D alkylation

N-

R3R 4

R

2 R 3 R R 2 (g) A profic acid and aldehyde solution is commonly added to an aqueous solution of compound The most preferred protic acid is formic acid. "he mcst preferred aldehyde is formaldehyde. The artisan can readily choose other appropriate reagents to facilitate the reductive alkylation. The resulting solution is refluxed for a period of about 4 to 80 hours. After reflux the solution should be made basic using an appropriate base such as potassium carbonate. The desired product can then be extracted with an appropriate organic phase, such as chloroform. The product WO 95/24200 PCT/US95/03099 48 can be dried, concentrated, and purified by known methods such as flash chromatography.

A preferred method for preparing certain Formula I compounds, wherein R 2 is hydrogen, utilizes the modified Pictet-Spengler reaction described supra, as represented by Scheme V. The substituents are as defined hereinabove.

Scheme V o 2 Merotic A A7R 4 acid

NH

S4 Me N R R4

H

Compound and compound are contacted in a suitable protic aqueous acid solution. When compounds having hydrogen at the 1-position are desired, glyoxylic acid may be used in place of This step may be completed under inert conditions. Compound and compound may be refluxed under atmospheric or inert conditions for a period of about to about 30 hours. Preferred protic acids include sulfuric acid and hydrochloric acid. The most preferred acid solution is 1 N HC1. If direct isolation is not effective, then the reaction mixture may be neutralized with an appropriate base, such as potassium carbonate, followed by extraction with an organic phase, such as chloroform. The product can be isolated through solvent removal followed by chromatographic isolation, such as silica gel chromatography, or other common isolation techniques. Typically the product is isolated as the acid addition salt. Appropriate salt forms are discussed supra.

As noted above, the compounds of the present invention can exist as resolved enantiomers. The single (-)enantiomer may be prepared by the chemical resolution method of A.I. Meyers as represented by Scheme VI infra. The (+)enantiomer may be prepared using known resolution WO 95/24200 PCT/US95/03099 49 techniques described supra. All substituents are as defined hereinabove.

Scheme VI Me Me NH ,o0tBu Me Me H xvlenes A

I

Me2N CSA N/f N OtBu

H

Me Me (M) A 1 1 6 (m) N N N OtBu

H

KH, THF,

TMEDA,

MOMC1 Me Me Me Me c^ Not'/ ci H Ka

N

N N OtBu n-RT, AI I A4 THF G N N OMe O R4 (o) OMe reduce

H

2

NNH

2 ,acid solvent a 1 I NH

N

H R4 (p) In Scheme VI, CSA represents camphorsulfonic acid.

Butylformadine is prepared from the amino acid valine using known methods. Other formadine compounds will also work. In step 1, the compound and butylformadine (1) solution is refluxed for a period of about 70 to 80 hours.

The product of the reflux reaction can be purified by standard isolation methods, such as flash chromatography.

The isolated oil can be used without further purification.

WO 95/24200 PCT/US95/03099 Compound prepared in step 1, can be added to a suspension of potassium hydride (iH) in tetrahydrofuran (THF). Tetramethylethylenediamine (TMEDA) and then chloromethylmethyl ether (MOMC1) are added to the solution, as represented by step 2. The mixture is stirred for a period of about 1 hour. The mixture can be treated with water and partitioned between an appropriate organic, such as diethyl ether, and water. The product should be extracted with the organic phee, dried over potassium carbonate, and concentrated. The isulting oil may be used in subsequent steps without further purification.

In step 3, n-BuLi is slowly added dropwise to a stirred, c sled (about -76 0 C to -80 0 C) solution of the formadine in dry THF. The solution is stirred for a period of about 1 hour followed by addition of the chloro compound in dry THF, The solution is stirred for an additional period of about 4-5 hours at the reduced temperature. The mixture is allowed to cool to room temperature for a period of about 4 to 14 hours. Wet THF is added and the solution concentrated. The residue is dissolved in an appropriate organic solvent such as chloroform and washed with water.

The organic phase is dried over a suitable drying agent, such as sodium carbonate, and concentrated to facilitate purification of the desired product. The product may be isolated by flash chromatography and concentrated. The resulting oil may be used in subsequent steps without further purification.

The deprotection reaction represented in otep 4 is begun at reduced temperature (about D0C). Water, acetic acid, and hydrazine hydrate are added to compound The reaction temperature is decreased to about -10 0 C to -20 0 C for a period of about 60-120 hours. The mixture is allowed to warm to ambient temperature and is concentrated. The product is dissolved in an appropriate organic phase, such as chloroform, and washed with water. The organic phase is dried over a suitable drying agent, such as sodium carbonate, and concentrated to a viscous oil. The oil is dissolved in WO 95/24200 PCT/US95/03099 51 an appropriate solvent, such as diethyl ether and treated with a suitable organic or inorganic acid to afford the desired acid addition salt. The salt can be isolated and purified by common chemical methods.

If the desired product has an alkyl group at the R 3 position, the reaction represented by Scheme VII may be employed.

Scheme VII (Step a) Ai I NH 2 HCi A 1 o base, solvent d I phthalic anhydride H PhCH 3 r base, THF,

R

3 halo, TMEDA NH2HCI A NH 2 HC1 H 2

NNH

2 alcohol A 0 R, (t) In Scheme VII, an appropriate saturated base solution, such as sodium carbonate, is added to compound The desired compound salt may be prepared by the method of Scheme II, above. The mixture is stirred at about ambient temperature for a period of about 1 hour. The layers are separated, and the aqueous layer is extracted with an appropriate organic solvent, such as chloroform. The organic layers are dried over an appropriate drying agent, such as sodium sulfate, and concentrated. The residue is dissolved in a suitable solvent such as toluene and treated with phthalic anhydride. The solution is refluxed for a period of about 12 to 20 hours with azeotropic drying. The solution is cooled, concentrated, and recrystallized to give compound WO 95/24200 PCT/US95/03099 52 In the next step, compound is mixed in THF. A cooled (about 0°C) suspension of an appropriate base, such as potassium hydride in dry THF, is slowly added to the compound solution. After the addition of the the base, the mixture is stirred for a period of about 1 hour.

Tetramethylethylenediamine (TMEDA) is added, followed by a haloalkyl such as methyl iodide (Mel). After about 1 hour, the reaction is quenched by the addition of water, followed by extraction with an appropriate organic phase, such as diethyl ether. The organic phases are dried over an appropriate drying agent, such as magnesium sulfate and concentrated.

The solution of the concentrated compound can be used directly in the next step. It is contacted with an appropriate solvent, such as methanol, and treated with hydrazine. The mixture is refluxed for a period of about 2 hours. The mixture is cooled to ambient temperature and treated with concentrated acid, such as HC1. The mixture is then treated with an alcohol and refluxed for a period of about 12 to 20 hours. Preferred alcohols include methanol, ethanol, and butanol. After cooling to ambient temperature, the mixture is partitioned between a suitable organic and an aqueous phase. One suitable combination is chloroform and concentrated sodium carbonate solution. The aqueous layer may be further extracted, the organic phases combined, dried, and concentrated. The product may be purified by flash chromatography, concentrated, and converted to a desired salt. The resulting compound may be used in Scheme III or Scheme V to produce the desired Formula I compound.

Compounds of Formulas XI and XII may be prepared using the methods described supra. However, a preferred method for preparing compounds of Formula XI and XII is illustrated by Scheme VIII WO 95/24200 WO 9524200PCTIUS95/03099 Scheme Vill 1 .R'2MgBr Z 2.R 32 ,S.CuBr N R30 R 3 1 H 3. W

N

Boc 4.acid

R

0

R

3 1 A NH 2 *acid

N

H

0 or glyoxylic acid mild acid R39

R

31 J~N-acid N Q~

H

wherein R 32 is independently selected from Cl-C 6 alkyl; A, and Q1 are defined supra.

Further, compounds of Example 108 can be prepared as illustrated by the following Scheme: M P Me Mao 0~ Ma. M e MeMgBr M Nk M 2.Me 2 5.CuBr P' NHVHCI MaO:0 Me NH-HCI MnN Ma. Ne e N H 3.H H laN 2d laBoc 3h W~e 4. HCI Wea 108 Similarly, compounds of Example 109 can be prepared as illustrated by the following Scheme: 0 Me MeDOe N0e

M

Me

~NH

2 *HCI 0 I Me0

H

31(109) 4c!1110) The following Examples further illustrate the preparation of certain of the Formula 1, 11, X7, and XII compounds. The examples are illustrative only, and are not intended~ to limit the scope of the invention.

The column chromatography procedures used standard flash chromatography techniques. One well-known reference WO 95/24200 PCT/US95/03099 54 describing appropriate flash chromotagraphy techniques is Still, W.C. Kahn, and Mitra, J. Org. Chem. 1978, 43, 2932.

Fractions containing product were generally evaporated under reduced vacuum to provide the product.

Optical rotations were obtained using methanol, pyridine, or other suitable solvent.

The hydrochloride salt of the particular compound was prepared by placing the free base into diethyl ether containing an alcohol such as methanol or other suitable solvent mixture. While stirring this ether solution, a solution of HC1 in diethyl ether was added dropwise until the solution became acidic. Alternatively, the ether solution was treated with dry HC1 gas.

The maleate salt of the particular compound was prepared by placing the free base in ethyl acetate or other suitable solvent and treating with maleic acid. The precipitate formed was filtered and dried to provide the corresponding hydrochloride or maleate salt of the free base.

Compounds of Formulas I through VI and VIII through XI± are more preferred for treating a mammal suffering from or susceptible to a condition associated with abnormal or dysfunctional 5-HT2B receptor stimulation.

Additionally, compounds of Formulas I through VI and VIII through XII are more preferred for blocking a 5-HT2B receptor in a mammal or in vitro. Finally, compounds of Formulas I through VI and VIII through XII are more preferred for use in an article of manufacture.

WO 95/24200 WO 9524200PCT/US9S103099 lox~jI 1. MeMgBr H 2.Me 2 S*CuBr 1t 2al3O=-3 N 2bn 3 0 4 Boo 2c n 30 5 N Me H o r gyoxyl ic a cida 4" 1N HCI a X"=5-Me b X"=7-CI c X'=5,7-diMe 4.HC1 a X"=5-Me n 30 -3 (90) b X"=7-CI n 30 (91) c X"=-5,7-Mep n 30 -3 d X"=5-Me n 30 -4 (92) e X'=7-CI n 30 -4 (93) f X"-=5,7-Me 2 n 30 -4 (94) g X"=5-Me n 30 -5 a X"=5-Me n 30 =4 R, R'=H b X'=7-CI n 30 =4 R, R'=H (96) c X"=5-Me n 30 =3 R-3,4-(OMe) 2 Bn, R=H (97) d X"=7-CI n 3 1=3 R-3,4-(QMe) 2 Bn, R'=H (98) eX"=5,!-M6 2 2 LJ, (9 f X"=5-Me n 30 =4 R-3,4(OMe) 2 Bn, R'=H (100) g Xlv=7-CI n 30 =4 R-3,4-(OMe) 2 Bn, R'=H (101) h X"=5,7-Me 2 n 30 -4 R=3,4-(OMe) 2 Bn, R'=H(1 02) I X"=5-Me n 30 -5 R-3,4-(OMe) 2 Bn, R'=H (103) 1 X"=5-Me n 3 9 0 =3 R=1-naphthylmethyl, R'=H (104) k X"=5-Me n 30 =4 R-l-naphthylmethyl, R'=H (105) I X"=5,7-Me 2 n 30 =4 R=1l-naphthylmethyl, (106) m X"=5-Me ri 30 *4 R, H (107) Q~ OMe OMe For Examples 90 through 109, where applicable, diethylether was distilled from sodium benzophenone ketyl prior to use. All reactions were performed under a positive pressure of argon. IH-NMR and 13 C-NMR data were recorded on a Bruker AC-200P (200 MHz). IR spectra were obtained on Nicolet 510 P-FT (film and KBr) Melting points were determined on a Balchi apparatus and are not corrected. Analytical TLC was performed on Merck TLC glass plates precoated with F254 silica gel 60 (UV, 254 nin and Iodine) Chromatographic separations were performed by using 230-400 mesh silica gel (Merck). N-BOC-aziridines (2a-d) were prepared from the corresponding alkenes following standard procedures.

WO 95/24200 PCT/US95/03099 56 Preparation 1 Preparation of 4-chlorobutanal.

O O CI% Hi,5% Pd/C CI c THF,2, 6-lutidin, psi 4-Chlorobutyryl chloride (300 g, 2.13 mol.) was dissolved in dry THF (3 To this solution was added 2,6lutidine (252 mL) followed by 5% Pd/C (30 This mixture was placed in a Parr hydrogenator and shaken under 60 psi of hydrogen for 6 hours. The mixture was purged with nitrogen, filtered, washing the catalyst with THF (500 mL), and concentrated at room temperature under reduced pressure.

Distillation afforded 4-chlorobutanal (148.3 g) as a colorless liquid.

Example 1 Preparation of 8-methyl-1-[(3,4-dimethoxyphenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole CHO N NMeO Sc 2 H 0 MeO Ac2, NaOAc. 100'C MeO

N

MeO I le OMe Me A solution of 3,4-dimethoxybenzaldehyde (24.5 g, 0.15 mol.), N-acetylglycine (17.4 g, 0.15 mol.) and sodium acetate (12.1 g, 0.15 mol) in acetic anhydride (135 mL) was heated to 100 0 C for 12 hours. The reaction mixture was cooled to ambient temperature poured onto ice (300 mL) with stirring. The product was isolated by filtration, washed with water (3 X 50 mL) and diethyl ether (3 X 50 mL) and dried under reduced pressure (16.3g).

WO 95/24200 PCT/US95/03099 57

NH

2 HC1 NH maleate 1 N HCI, reflux H O KCOa: N ai\ ae acid H Me Me m MeOe OMe MeO OMe A suspension of azalactone prepared above (1.35 g, 5.46 mmol.) and 7-methyl-tryptamine hydrochloride (1.15 g, 5.46 mmol.) in 1 N HC1 (50 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% NH40H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid.

The product was isolated as the maleate salt (730 mg) by filtration. (mp 168 0 C, dec.) Analysis Calculated Found C 66.36 66.15 H 6.24 6.28 N 6.19 5.79 WO 95/24200 PCT/US95/03099 58 Example 2 Preparation of 8-bromo-l-[(3,4-dimethoxyphenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride To a stirred suspension of 2-bromophenyl-hydrazine hydrochloride (25.8 g, 115 mmol.) in chloroform (500 mL) was added saturated sodium carbonate solution (500 mL). The mixture was stirred for 30 minutes and extracted with chloroform (2X 200 mL). The combined organic phases were concentrated to afford the hydrazine free base as a yellow oil. This oil was dissolved in methanol (100 mL) and treated slowly with 4-chlorobutanal (12.3 g, 115 mmol). The mixture was placed in a sealable tube and purged with nitrogen for minutes. The tube was sealed and placed in an oil bath preheated to 95°C. Heating was continued for 18 hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were ccbined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HCI gas. The hydrochloride salt was isolated by filtration, washed with 2-propanol (50 mL) and diethyl ether (100 mL) and dried to afford 7-bromotryptamine hydrochloride (3.6 g) as a pale solid, which was used without further purification.

A suspension of azalactone (prepared as described in Example 1) (1.16 g, 4.7 mmol.) and 7-bromotryptamine hydrochloride (1.0 g, 3.6 mmol.) in 1 N HC1 '100 nmL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The brown solid was triturated with isopropyl alcohol WO 95/24200 PCT/US95/03099 59 (3 X 50 mL) and washed with diethyl ether (3 X 50 mL).

Recrystallization from ethanol afforded 860 mg of desired product as the hydrochloride salt. (mp 279-281°C, dec.) Analysis Calculated Found C 54.87 54.75 H 5.07 5.20 N 6.40 6.23 Example 3 Preparation of 6,8-dibromo-l-[ (3,4-dimethoxyphenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b] indole To a stirred, cooled (-5 0 C) solution of 2,4dibromoaniline (50.0 g,0.2 mol.) in concentrated HC1 solution (110 mL) was added sodium nitrite (13.8 g, 0.2 mol.) in water (110 mL) dropwise at such a rate as to maintain temperature below 5 0 C. After complete addition, the mixture was further stirred at 5 0 C for 30 minutes. A solution of tin chloride monohydrate (135.4 g, 0.6 mol.) in concentrated HC1 (total volume 170 mL) was added dropwise again maintaining temperature below 5 0 C. After complete addition and minutes of further stirring, -the mixture was placed in the freezer overnight. The light brown solid which precipitated was isolated by filtration and washed with cold brine followed by a solution of petroleum ether/ diethyl ether (2/1 by volume). This solid was slowly added to an ice cooled mixture of 50% sodium hydroxide solution/ ethyl acetate. The mixture was extracted with ethyl acetate and the organic phase dried over magnesium sulfate. After filtration, the solution was concentrated to 400 mL total volume diluted with diethyl ether (1.5 L) and treated with dry HC1. The product, 2,4-dibromophenyl-hydrazine hydrochloride (45.9 g) was isolated as a white solid and used without further purification.

To a stirred suspension of 2,4-dibromophenylhydrazine hydrochloride (22.0 g, 83 mmol.) in chloroform (500 WO 95/24200 PCT/US95/03099 mL) was added saturated potassium carbonate solution (500 mL). The mixture was stirred for 30 minutes and extracted with chloroform (2X 200 mL). The combined organic phases were concentrated to afford the hydrazine free base as a yellow oil. This oil was dissolved in methanol (163 mL) and treated slowly with 4-chlorobutanal (8.8 g, 83 mmol.). The mixture was placed in a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oil bath preheated to 95 0 C. Heating was continued for 18 hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were combined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HC1 gas. The hydrochloride salt was isolated by filtration, washed with 2-propanol mL) and diethyl ether (100 mL) and dried to afford 7-bromotryptamine hydrochloride (1.5 g) as a pale solid, which was used without further purification.

A suspension of azalactone (prepared as described in Example 1) (0.45 g, 1.82 mmol.) and 5,7-dibromotryptamine hydrochloride (0.58 g, 1.64 mmol.) in 1 N HC1 (65 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqaeous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as he maleate salt (340 mg) by filtration. (mp 177-179 0 C, dec.) WO 95/24200 PCT/US95IO3099 Analysis

C

H

N

Calculated 48.34 4.06 4.70 Found 48.61 4 .17 4.69

I

WO 95/24200 PCT/US95/03099 62 Eample 4 Preparation of 6-methyl-8-bromo-l-[(3,4-dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride To a stirred, cooled (-5 0 C) solution of 2-bromo-4methylaniline (50.54 g, 0.272 mol.) in concentrated HC1 solution (200 was added sodium nitrite (18.9 g, 0.274 mol.) in water (200 mL) dropwise at such a rate as to maintain temperature below 5 0 C. After complete addition, the mixture was further stirred at 50C for 30 minutes. A solution of tin chloride monohydrate (185.4 g, 0.822 mol.) in concentrated HC1 (total volume 400 mL) was added dropwise again maintaining temperature below 5 0 C. After complete addition and 30 minutes of further stirring, the mixture was placed in the freezer overnight. The light brown solid which precipitated was isolated by filtration and washed with cold brine followed by a solution of petroleum ether/diethyl ether (2/1 by volume). This solid was slowly added to an ice cooled mixture of 50% sodium hydroxide solution ethyl acetate. The mixture was extracted with ethyl acetate and the organic phase dried over magnesium sulfate. After filtration, the solution was concentrated to 400 mL total volume diluted with diethyl ether (1.5 L) and treated with dry HC1. The product, 2-bromo-4-methylphenylhydrazine hydrochloride (52.4 g) was isolated as a light brown solid and used without further purification.

5-Methyl-7-bromotryptamine hydrochloride (4.95 g) was prepared as described in Example 3, except using 2-bromo-4-methylphenyl hydrazine hydrochloride (21 g) as starting material.

A suspension of azalactone (prepared as described in Example 5) (1.44 g, 6.07 mmol.) and 5-methyl-7bromotryptamine hydrochloride (1.12 g, 3.87 mmol.) in 1 N HCI mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration.

The brown solid was triturated with isopropyl alcohol (3 X i WO 95/24200 PCT/US95/03099 63 mL) and washed with diethyl ether (3 X 50 mL).

Recrystallization from ethanol afforded 1.06 g of desired product as a pale solid. (mp 251-253 0 C, dec.) Analysis Calculated Found C 55.83 56.08 H 5.35 5.32 N 6.20 6.33 Example Preparation of 8-methoxy-l-[(3,4-dimethoxyphenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole To a stirred, cooled (0 0 C) suspension of 2methoxyphenylhydrazine hydrochloride (14.44 g, 83 mmol.) in THF (600 mL) was added 4-chlorobutanal (9.0 g, 84 mmol.) followed by dropwise addition of triethylamine (8.6 g, mmol.) in THF (20 mL) Upon complete addition, the cooling bath was removed and the solution stirred for 1 hour. The reaction mixture was filtered and the filter cake washed with THF (100 mL). The combined filtrates were concentrated to an orange oil, which was dissolved in methanol (150 mL) and water (5 mL). The solution was transferre. to a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oilbath preheated to 95 0 C. After heating for 14 hours, the reaction mixture was cooled to ambient temperature and concentrated under reduced pressure.

The residue was partitioned between saturated aqueous potassium carbonate and 3:1 chloroform: 2-propanol. The organic phase was dried over sodium sulfate and concentrated.

The residue was purified by flash chromatography on silica gel (15% methanol, 0.2% NH 4 0H, in chloroform as eluen.). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in methanol and treated with dry HCt and concentrated to afford 7-methoxytryptamine hydrochloride (4.04 g) as a stable foam, which was used without further purification.

WOr 95/124200 PCT/US95/03099 64 A suspension of azalactone (prepared as described in Example 1) (1.20 g, 4.85 mmol.) and 7-methoxytryptamine hydrochloride (1.0 g, 4.4 mmol.) in 1N HC1 (120 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutral, ted with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (770 mg) by filtration. (mp 219-220 0 C, dec.).

Analysis Calculated Found C 64.09 64.04 H 6.02 6.18 N 5.98 5.93 Example 6 Preparation of 6,8-difluoro-l- (3,4-dimethoxyphenyl)-methyl 1,2,3,4-tetrahydro-9H-pyrido[,4-b]indole L HNH2 HC1

NCO

3

CHCI

3 F NH 2 uCi NHNH2 HC1 K2C0 CHC1z; MeOH,NaOAc, oC; N C1 Et20, MeOH, HCI H

F

0 To a stirred suspension of 2,4-difluorophenylhydrazine hydrochloride (18.5 g, 128 mmol.) in chloroform (500 mL) was added saturated potassium carbonate solution (500 mL). The mixture was stirred for 30 minutes and extracted with chloroform (2X 200 mL). The combined org'nic phases were concentrated to afford the hydrazine free WO 95/24200 PCT/US95/03099 base as a yellow oil. This oil was dissolved in a solution of methanol (163 mL), water (36 mL) and sodium acetate (10.57 g) and treated slowly with 4-chlorobutanal (13.7 g, 128 mmol.). The mixture was placed in a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oil bath preheated to 95 0 C. Heating was continued for hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were combined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HC1 gas. The hydrochloride salt was isolated by filtration, washed with 2-propanol mL) and diethyl ether (100 mL) and dried to afford 7 bromotryptamine hydrochloride (6.3 g) as a pale solid, which was used without further purification.

NH

2 HC1 NH maleate 1 N HCI, reflux lkl N 0

M.

N 0 2

N

r% 2

CO

3

N

I mallcadd H Fm Me

N

MeO OMe MeO

O

OMe A suspension of azalactone (prepared as described in Example 1) (1.07 g, 4.33 mmol.) and 5,7-difluorotryptamine hydrochloride (1.0 g, 4.3 mmol.) in 1 N HC1 (70 mL) was heated to reflux for 65 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate WO 95/24200 PCT/US95/03099 66 solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and -he residue chromatographed on silica gel (ethyl acetate 0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (450 mg) by filtration. (mp 164-166 0 C, dec.) Analysis Calculated Found C 60.76 60.63 H 5.10 5.14 N 5.90 5.82 Examp le 7 Preparation of 7-methyl-8-bromo-l-[(3,4-dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride 2-Bromo-3-methylphenylhydrazine hydrochloride (23 g) was prepared as described for 2-bromo-4methylphenylhydrazine hydrochloride in Example 4, except using 2-bromo-3-methylaniline as starting material.

6-Methyl-7-bromotryptamine hydrochloride was prepared (2.42 q) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 2-bromo-3methylphenylhydrazine hydrochloride as starting material.

A suspension of azalactone (prer-red as described in Example 1) (3.63 g, 14.7 mmol.) and 6-r yl-7-bromotryptamine hydrochloride (4.25 g, 4.21 mmol.) in 1 N HCI (150 mL) was heated to reflux for 18 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% NH40H as eluent). The fractions containing product were pooled and concentrated under reduced pressure.

WO 95/24200 PCT/US95/03099 67 The residue was dissolved in ethyl acetate containing 1% methanol and treated with dry HC1. The product was isolated as the hydrochloride salt (3.11 g) by filtration. m/e=414.

Analysis Calculated Found C. 55.83 56.13 H 5.18 5.29 N 6.20 6.31 Example 8 Preparation of 6-(1,l-dimethylethyl)-1-[(3,4-dimethoxyphenyl) methyl]1,2,3,4-tetrahydro-1-9H-pyrido-[3,4b]indole hydrochloride 1-dimethylethyl) -tryptamine hydrochloride was prepared (2.95 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 4-(1,1dimGthylethyl)-phenylhydrazine hydrochloride (6.00 g) as starting material.

A suspension of azalactone (prepared as described in Example-1) (1.25 g, 5.26 mmol.) and 5-(1,1-dimethylethyl)tryptamine hydrochloride (1.33 g, 5.26 mmol.) in 1 N HC1 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, and the crude product isolated by filtration.

The brown solid was triturated with isopropyl alcohol (3 X mL) and washed with diethyl ether (3 X 50 mL).

Recrystallization from ethanol afforded 0.74 g of desired product as a pale solid.

Analysis Calculated Found C 69.47 69.66 H 7.53 7.50 N 6.75 6.71 Example 9 Preparation of 5-fluoro-6-methyl-1-[(3,4-dimethoxyphenyl)methyl] -1,2,3,4-tetrahydro-9H-pyrido[3,4-b] indole WO 95/24200 PCT/US95/03099 68 3-Fluoro-4-methylphenylhydrazine hydrochloride (21.4 g) was prepared as described for 2-bromo-4 methylphenylhydrazine hydrochloride in Example 4, except using 3-fluoro-4-methylaniline as starting material.

hydrochloride was prepared (2.20 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 3-fluoro-4methylphenylhydrazine hydrochloride (6.00 g) as starting material.

A suspension of azalactone (prepared as described in Example 1) (0.76 g, 3.06 mmol.) and methyltryptamine hydrochloride (0.70 g, 3.06 mmol.) in 1 N HC1 (40 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure.

The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (60 mg) by filtration. mp. 191- 194 0

C.

Analysis Calculated Found C 63.82 63.60 H 5.78 5.65 N 5.95 5.92 Example Preparation of 7,8,9,10-tetrahydro-10-[(3,4-dimethoxyphenyl) methyl]-11H-benzo[g]pyrido[3,4-b]indole 6,7-Benzotryptamine hydrochloride was prepared (2.85 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using l-naphthyl-hydrazine hydrochloride (6.00 g) as starting material.

WO 95/24200 PCTUS95/03099 69 A suspension of azalactone (prepared as described in Example 1) (1.51 g, 6.11 mmol.) and 6,7-benzotryptamine hydrochloride (1.50 g, 6.11 mmol.) in 1 N HC1 (40 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (240 mg) by filtration. m/e=373, mp. 187 0

C

(dec.) Analysis Calculated Found C 68.84 68.63 H 5.78 5.91 N 5.73 5.67 Example 11 Preparation of 6-cyclohexy-l-1[(3,4-dimethoxyphenyl)-methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride 4-Cyclohexylphenylhydrazine hydrochloride (35.6 g) was prepared as described for 2-bromo-4-methylphenylhydrazine hydrochloride in Example 4, except using 4cyclohexylaniline as starting material.

hydrochloride was prepared (1.29 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 4cyclohaxylphenylhydrazine hydrochloride as starting material.

A suspension of azalactone (prepared as described in Example 1) (0.54 g, 2.18 mmol.) and hydrochloride (0.6 g, 2.18 mmol.) in 1 N HC1 (30 mL) was heated to reflux for 14 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, WO 95/24200 PCT/US95/03099 neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate-containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (140 mg) by filtration. m/e= 404.

Analysis Calculated Found C 69.21 69.17 H 6.97 7.01 N 5.38 5.53 Example 12 Preparation of 5,8-dimethyl-1,2,3,4-tetrahydro--[ (3,4dimethoxyphenyl)methyl] -9H-pyrido[3,4b] indole hydrochloride 4,7-dimethyltryptamine hydrochloride was prepared (0.94 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using phenylhydrazine hydrochloride (16.8 g) as starting material.

A suspension of azalactone (prepared as described in Example 1) (1.04 g, 4.21 mmol.) and 4,7-dimethyltryptamine hydrochloride (0.94 g, 4.21 mmol.) in 1 N HC1 (40 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with anhydrous HC1. The product was isolated as the hydrochloride salt (370 mg) by filtration. m/e=349 WO 95/24200 PCT/US95/03099 71 Analysis Calculated Found C 68.29 68.59 H 7.03 6.92 N 7.24 7.04 Example 13 Preparation of 6-(l-metethethyl)-1,2,3,4-tetrahydro--[(3,4dimethoxyphenyl)methyl]-9H-pyrido[3,4b]indole To a stirred suspension of 4-isopropylphenylhydrazine hydrochloride monohydrate (15.3 g, 91.95 mmol.) in chloroform (250 mL) was added saturated sodium carbonate solution (250 mL). The mixture was stirred for 30 minutes and extracted with chloroform (2X 200 mL). The combined organic phases were concentrated to afford the hydrazine free base as a yellow oil. This oil was dissolved in methanol (200 mL) and water (5 mL) and treated with sodium acetate (6.72 g, 82 mmol.) and 4-chlorobutanal (8.7 g, 82 mmol.).

The mixture was placed in a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oil bath preheated to 100 0 C. Heating was continued for 18 hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were combined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HCI gas. The hydrochloride salt was isolated by filtration, washed with 2-propanol mL) and diethyl ether (100 mL) and dried to afford isopropyltryptamine hydrochloride (9.8 g) as a pale solid, which was used without further purification.

A suspension of azalactone (prepared as described in Example 1) (1.55 g, 6.31 mmol.) and I- WO 95/74200 PCT/US95/03099 72 hydrochloride (1.76 g, 7.37 mmol.) in 1 N HC1 (40 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic ;'cid. The product was isolated as the maleate salt (310 mg) by filtration. m/e=365, mp. 196-200 0

C.

Analysis Calculated Found C 67.48 67.74 H 6.71 6.75 N 5.83 5.92 Example 14 Preparation of 6,8-dimethyl-1,2,3,4-tetrahydro-l-[(3,4dimethoxyphenyl)methyl] -9H-pyrido [3,4b indole hydrochloride 5,7-Dimethyltryptamine hydrochloride was prepared (2.86 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 2,4dimethylphenylhydrazine hydrochloride (15.0 g) as starting material., A suspension of azalactone (prepared as described in Example 1) (1.65 g, 6.67 mmol.) and 5,7-dimethyltryptamine hydrochloride (1.50 g, 6.67 mmol.) in 1 N HC1 (70 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The react on mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol/hexanes (3 X 50 mL) and washed with hexanes (3 X 50 mL). The product was isolated by filtration (820 mg). m/e=350.

WO 95/24200 WO 9524200PCT/US9/03099 73 Analysis Calculated Found C 68.29 68.07 H 7.03 7.12 N 7.24 7.23 Example Preparation of 5, 7-dimethyl-1,2, 3,4-tetrahydro-1- dimethoxyphenyl )methyl] -9H-pyrido(3 ,4bJ indole hydrochloride 4, 6-Dimethyltryptamine hydrochloride was prepared (1.06 g) as described for S-methyl-7-bromotryptamine hydrochloride in Example 4, except using dimethylphenylhydra zine hydrochloride (7.65 g) as starting material.

A suspension of azalactone (prepared as described in Example 1) (1.16 g, 4.69 =mo1.) and 4,6-dimethyltryptamine hydrochloride (1.05 g, 4.67 mmol.) in 1 N HCl (60 mL) was heated to ref lux for 24 )urs under nitrogen atmosphere. .The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol/hexanes (3 X 50 mL) and washed with hexanes (3 X 50 mL) The product was isolated by filtration (770 mig) m/e=350.

Analysis Calculated Foundd C 68.29 68.09 H 7.03 7.12 N 7.24 7,02 Example 16 Preparation of 6,7-dimethyl-1,2,3,4-tetrahydro-1-[(3,4dimethoxyphel.) methyl] -9H-pyrido(3 ,4b] indole To a stirred, cooled (0 0 C) solution of 5,6dimethyl-indole (3.69 g, 25.4 mmol.) in dry diethyl ether WO 95/24200 PCT/US95/03099 74 mL) was added dropwise oxalyl chloride (3.8 mL, 43.0 mmol.) over 2 minutes. After further stirring for 30 minutes, the bright yellow acid chloride (5.99 g) was isolated by filtration and washed with dry diethyl ether. This acid chloride was added in portions to a rapidly stirred solution of aqueous ammonium hydroxide (100 mL). After the addition was complete, the mixture was further stirred at ambient temperature for 30 minutes and the crude product isolated by filtration. Recrystallization from THF/diethyl ether afforded product (3.05 g) as a tan solid.

To a stirred, refluxing solution of amide (prepared above) (3.05 g, 14.1 mmol.) in THF was added dropwise a suspension of lithium aluminum hydride (3.07 g, 81.3 mmol.) in THF over 1 H. Upon complete addition, the mixture was further heated to reflux for 14 H. The reaction mixture was cooled to 0°C and carefully treated with water (3.1 mL) followed by 15% sodium hydroxide solution (3.1 mL), followed by water (9.3 mL). The salts were removed by filtration and the filtrate was concentrated under reduced pressure. -The residue was dissolved in diethyl ether (80 mL) with 5% ethyl acetate and treated with anhydrous HC1. The hydrochloride salt (2.65 g) was isolated by filtration and washed with dry ether.

A suspension of azalactone (prepared as described in Example 1) (1.10 g, 4.45 mmol.) and 5,6-dimethyltryptamine hydrochloride (1.00 g, 4.45 mmol.) in 1 N HC1 (60 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixLure was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform.

The combined organic layers were concentrated under reduced pressure and the residue chromatographed on sili'.a gel (ethyl acetate 0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure.

The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was WO 95/24200 PCT/US95/03099 isolated as the maleate salt (450 mg) by filtration. mp. 197- 200 0

C.

Analysis Calculated Found C 66.94 67.01 H 6.48 6.56 N 6.00 5.98 Example 17 Preparation of 6-ethyl-1,2,3,4-tetrahydro-l-[(3,4dimethoxyphenyl) methyl] -9H-pyrido 3,4b] indole To a stirred, cooled solution of ethylindole (4.0 g, 27.5 mmol.) in dry diethyl ether (250 mL) was added dropwise oxalyl chloride (4.8 mL, 55.1 mmol.) over 2 minutes. After further stirring for 30 minutes, the bright yellow acid chloride was isolated by filtration and washed with dry diethyl ether. This acid chloride was added in portions to a rapidly stirred solution of aqueous ammonium hydroxide (200 mL). After the addition was complete, the mixture was further stirred at ambient temperature for 30 minutes and the crude product isolated by filtration (4.7 g) as a tan solid.

To a stirred, refluxing solution of amide (prepared above) (4.7 g, 21.7 mmol.) in THF was added dropwise a suspension of lithium aluminum hydride (4.7 g, 121 mmol.) in THF over 1 H. Upon complete addition, the mixture was further heated to reflux for 14 H. The reaction mixture was cooled to 0 C and carefully treated with water (4.7 mL) followed by 15% sodium hydroxide solution (4.7 mL), followed by water (14.1 mL). The salts were removed by filtration and the filtrate was concentrated under reduced pressure. The residue was dissolved in diethyl ether (80 mL) with 5% ethyl acetate and treated with anhydrous HC1. The hydrochloride salt (4.02 g) was isolated by filtration and washed with dry ether.

A suspension of azalactone (prepared as described in Example 1) (1.10 g, 4.45 mmol.) and 5,6-dimethyltryptamine WO 95/24200 PCT/US95/03099 76 hydrochloride (1.00 g, 4.45 mmol.) in 1 N HC1 (60 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (520 mg) by filtration. mp. 1850C(dec.).

Analysis Calculated Found C 66.94 66.95 H 6.48 6.55 N 6.01 5,99 ExamDle 18 Preparation of 6-bromo-1,2,3,4-tetrahydro-l-[(3,4dimethoxyphenyl)methyl]-9H-pyrido[3,4b]indole A suspension of azalactone (prepared as described in Example 1) (0.91 g, mmol.) and hydrochloride (1.01 g, 3.7 mmol.) in 1 N HC1 (60 mL) was heated to reflux for 18 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% as eluent). The fractions containing product were pooled and concentratPd under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolate, as the maleate salt (800 mg) by filtration. (mp 184-188 0 C, dec.) m/e= 403.

c~ WO 95/24200 PCT/US95/03099 77 Analysis Calculated Found C 55.72 55.51 H 4.87 5.09 N 5.41 5.36 Example 19 Preparation of 7,8-dimethyl-1,2,3,4-tetrahydro-l- (3,4dimethoxyhenyl) methyl] -9H-pyrido [3,4b] indole hydrochloride 6,7-Dimethyltryptamine hydrochloride was prepared (2.26 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 2,2-dimethylphenylhydrazine hydrochloride (15.0 g) as starting material.

A suspension of azalactone (prepared as described in Example 1) (1.39 g, 5.62 mmol.) and 6,7-dimethyltryptamine hydrochloride (1.26 g, 5.61 mmol.) in 1 N HC1 (70 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2%

NH

4 0H as eluent). The fractions containing product were pooled and concentrated under'reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with anhydrous HC1. The product was isolated as the hydrochloride salt (290 mg) by filtration. m/e=350 Analysis Calculated Found C 68.29 68.51 H 7.03 6.87 N 7.24 7.22 Example WO 95/24200 PCT/US95/03099 78 Preparation of 6-methyl-1,2,3,4-tetrahydro-l-[(3,4dimethoxyphenyl)methyl]-9H-pyrido[3,4b]indole hydrochloride A suspension of azalactone (prepared as described in Example 1) (3.4 g, 12.4 mmol.) and hydrochloride (2.0 g, 9.9 mmol.) in 1 N HC1 (100 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol and washed with diethyl ether The product was isolated as WO 95/24200 PCT/US95/03099 79 the hydrochloride salt by filtration (3.2 mp. 245-2460C (dec.) Analysis Calculated Found C 67.64 67.42 H 6.67 6.66 N 7.51 7.25 Example 21 Preparation of 6-methyl-l-[(3,4,5-trimethoxyphenyl)-methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride A solution of 3,4,5-trimethoxybenzaldehyde (20.0 g, 0.10 mol.), N-acetylglycine (11.9 g, 0.10 mol.) and sodium acetate (8.4 g, 0.1 mol) in acetic anhydride (100 mL) was heated to 100 0 C for 2 hours. The reaction mixture was cooled to ambient temperature poured onto ice (300 mL) with stirring. The product was isolated by filtration, washed with water (3 X 50 mL) and diethyl ether (3 X 50 mL) and dried under reduced pressure (5.6 g).

A suspension of azalactone (prepared above) g, 7.2 mmol.) and 5-methyltryptamine hydrochloride (1.1 g, 5.4 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated Dy filtration. The solid was triturated with isopropanol and washed with diethyl ether The product was isolated by filtration (650 mg). mp. 228-229 0

C.

Analysis Calculated Found C 65.58 65.38 H 6.75 6.76 N 6.95 6.92 WO 95/24200 PCT/US95/03099 Example 22 Preparation of 6-methyl-- [(2,3,4-trimethoxyphenyl)-methyl] 1,2,3,4-tetrahydro-9H-pyrido indole hydrochloride Azalactone (12.28 g) was prepared as in Example 21 except using 2,3,4-trimethoxybenzaldehyde (20.0 g).

A suspension of azalactone (prepared above) g, 7.2 mmol.) and 5-methyltryptamine hydrochloride (1.1 g, 5.4 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with isopropanol and washed with diethyl ether The product was isolated by filtration (1.36 mp. 214.5 0

C.

Analysis Calculated Found C 65.58 65.41 H 6.75 6.70 N 6.95 6.89 Example 23 Preparation of 6-methyl-l- [(2-methoxyphenyl)methyl -1,2,3,4tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (16.42 g) was prepared as in Example 21 except using 2-methoxybenzaldehyde (20.0 g).

A suspension of azalactone (prepared above) g, 9.2 mmol.) and 5-methyltryptamine hydrochloride (1.5 g, 6.9 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with isopropanol and washed with diethyl ether The product was isolated by filtration (880 mg). mp. 252.8 0

C.

Analysis Calculated Found C 70.06 70.15 H 6.76 6.83 N 8.17 8.16 WO 95/24200 PCT/US95/03099 81 Example 24 Preparation of 6-methyl-l-[(2,4-dimethoxyphenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (7.55 g) was prepared as in Example 21 except using 2,4-dimethoxybenzaldehyde (20.0 g).

A suspension of azalactone (prepared above) g, 8.1 mmol.) and 5-methyltryptamine hydrochloride (1.3 g, 6.1 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with anhydrous HC1. The product was isolated as the hydrochloride salt (361 mg) by filtration. mp. 262.6 0

C.

Analysis Calculated Found C 67.64 67.73 H 6.76 6.85 N 7.51 7.50 Example Preparation of 6-methyl-1-[(2,5-dimethoxyphenyl)methyl 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (13.21 g) was prepared as in Example 21 except using 2,5-dimethoxybenzaldehyde (20.0 g).

A suspension of azalactone (prepared above) g, 8.1 mmol.) and 5-methyltryptamine hydrochloride (1.3 g, 6.1 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with WO 95/24200 PCT/US95/03099 82 chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH40H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with anhydrous HC1. The product was isolated as the hydrochloride salt (1.14 g) by filtration, mp. 262 oC.

Analysis Calculated Found C 67.64 67.36 H 6.76 6.71 N 7.51 7.25 Example 26 Preparation of 6-methyl-1-[(2,4,5-trimethoxyphenyl)-methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (8.36 g) was prepared as in Example 21 except using 2,4,5-trimethoxybenzaldehyde (20.0 g).

A suspension of azalactone (prepared above) g, 7.2 mmol.) and 5-methyltryptamine hydrochloride (1.1 g, 5.4 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with isopropanol and washed with diethyl ether The product was isolated by filtration. Recrystallization from ethanol/cyclohexane afforded product (299 mg). mp. 176.3 0

C.

Analysis Calculated Found C 65.58 65,51 H 6.75 6.73 N 6.95 6.87 WO 95/24200 PCT/US95/03099 83 Example 27 Preparation of 6-(1-methylethyl)-1-[(2,3,4-trimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride A suspension of azalactone (prepared as in Example 22) (1.0 g, 3.61 mmol.) and hydrochloride (prepared as in Example 13) (646 mg, 2.7 mmol.) in 1 N HC1 (20 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform.

The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure.

The residue was dissolved in ethyl acetate containing 1% methanol and treated with anhydrous HC1. The product was isolated as the hydrochloride salt (315 mg) by filtration.

mp. 147.3 0

C.

Analysis Calculated Found C 66.89 66.80 H 7.25 7.01 N 6.50 6.39 Example 28 Preparation of 6-methyl-- [(3,4-dimethoxy-5-nitrophenyl) methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (16.9 g) was prepared as in Example 21 except using 3,4-dimethoxy-5-nitrobenzaldehyde (23.5 g).

A suspension of azalactone (prepared above) (2.8 g, 9.6 mmol.) and 5-methyltryptamine hydrochloride (2.0 g, mmol.) in 1 N HC1 (50 mL) was heated to reflux for 72 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with isopropanol WO 95/24200 PCT/US95/03099 84 and washed with diethyl ether The product was isolated as the hydrochloride salt by filtration (3.44) mp. 239-243 0

C,

m/e=381.

Analysis Calculated Found C 60.36 60.54 H 5.79 5.66 N 10.06 10.12 Example 29 Preparation of 6-methyl-l-[(3-iodo-4,5-dimethoxy-phenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole To a stirred, cooled (0 0 C) solution of iodovanillin (10.0 g, 35.96 mmol.) in dimethylformamide (50 mL) was added anhydrous potassium carbonate (20.0 g, 143.86 mmol.) followed by iodomethane (3.11 mL, 50.0 mmol.). The mixture was allowed to warm to ambient temperature and stir for 14 H.

The mixture was poured into diethyl ether (500 mL) and washed with watpr (3 X 150 mL). The organic phase was dried over MgS0 4 and'concentrated under reduced pressure to afford 3- (9.5 g) as a yellow oil which solidified upon standing and was used without further purification.

Azalactone (11.1 g) was prepared as in Example 21 except using 3-iodo-4,5-dimethoxybenzaldehyde (9.5 and hippuric acid (6.41 g) instead of N-acetylglycine.

A suspension of azalactone (prepared above) (2.2 g, 5.0 mmol.) and 5-methyltryptamine hydrochloride (1.0 g, 4.3 mmol.) in 1 N HC1 (100 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous sodium hydroxide solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl WO 95/24200 PCT/US95/03099 acetate containing 1% methanol and treated with maleic acid.

The product was isolated as the maleate salt (134 mg) by filtration. m/e=463.

Analysis Calculated Found C 51.92 52.15 H 4.71 4.72 N 4.84 4.70 Example Preparation of 6-methyl-l-[(3,4-dimethoxy-5-amino-phenyl)methyl]-l,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole dihydrochloride To a stirred solution of nitro compound (prepared in Example 28) (3.0 g, 7.2 mmol.) in acetic acid (40 mL) was added activated zinc dust (4.64 The reaction mixture was stirred at ambient temperature for 2 H, diluted with water (200 mL) and filtered through celite. The filtrate was neutralized with aqueous ammonium hydroxide solution and extracted with chloroform. The organic phase was washed with brine and dried over magnesium sulfate. The combined organic ph is were concentrated under reduced pressure and the residue dissolved in ethyl acetate and treated with anhydrous HC1. The product was isolated by filtration, washing with diethyl ether and trituration with ethyl acetate to afford product as the bis-hydrochloride salt (2.41 g).mp. 230-234 0

C,

m/e=351.

Analysis Calculated Found C 59.44 58.47 H 6.41 6.31 N 9.90 9.68

M

WO 95/24200 PCT/US95/03099 86 Example 31 Preparation of 6-methyl-l-[(3-methoxy-4-propoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole To a stirred solution of vanillin (30.0 g, 197 r.mol.) in methanol (100 mL) was added anhydrous potassium carbonate (13.7 g, 99 mmol.) followed by allyl bromide (17.0 mL, 197 mmol.). The mixture was heated to reflux for 5 H.

The reaction mixture was filtered and concentrated under reduced pressure to afford the intermediate product (30.4 g) as an oily solid which was used without further purification.

Azalactone (32.2 g) was prepared as in Example 21 except using 3-methoxy-4-allyloxybenzaldehyde (30.4 and hippuric acid (28.3 g) instead of N-acetyl glycine.

A suspension of azalactone (prepared above) (1.74 g, 5.2 mmol.) and 5-methyltryptamine hydrochloride (1.1 g, 5.2 mmol.) in 1 N HCI (40 mL) and ethanol (30 mL) was heated to reflux for 18 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (560 mg) by filtration. m/e=362. The product was used without further purification.

To a suspension of maleate salt (560 mg, 1.7 mmol.) in chloroform (100 mL) was added saturated potassium carbonate solution (100 mL) with vigorous stirring. The layers were separated and the aqueous phase was further extracted with chloroform (2 Z 100 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The free base was dissolved in ethanol and hydrogenated (25 0 C, 60 PSI) in the presence of raney nickel catalyst. The catalyst was removed WO 95/24200 PCT/US95/03099 87 by filtration and the solution concentrated under reduced pressure to afford a viscous oil, which was dissolved in ethyl acetate and treated with maleic acid (140 mg). The crude product was isolated by filtration. Trituration with hot ethyl acetate and washing with diethyl ether afforded product (170 mg) as the maleate salt. mp. 188 0 C, m/e=365.

Analysis Calculated Found C 67.48 67.62 H 6.71 6.66 N 5.83 5.80 Example 32 Preparation of 6-methyl-l-[(4-dimethylaminophenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole dihydrochloride To a stirred, cooled (-78 0 C) suspension of me hoxymethyltriphenylphosphonium chloride (13.79 g, 40.02 mmol.) in dry THF (150 mL) was added n-BuLi solution (25.2 mL. 1.6 M, 40.02 mmol.) dropwise by syringe. The orange suspension was stirred at -78 0 C for 15 min. A solution of 4-dimethylaminobenzaldehyde (5.00 g, 3.35 mmol.) in THF mL) was added to the ylide dropwise over 10 min. The reaction mixture was gradually warmed to ambient temperature and stirred 14 H. Saturated ammonium chloride solution (100 mL) was added and the mixture extracted with diethyl ether (3 X 50 mL). The combined organic phases were dried over sodium sulfate and concentrated under reduced pressure.

Chromatography on silica gel, eluting with 15% ethyl acetate/hexanes afforded product (4.70 g) as a mixture of olefin isomers which was used without further purification.

A mixture of 5-methyltryptamine hydrochloride (891 mg, 4.23 mmol.) and l-methoxy-4'-dimethylaminostyrene (1.00 g, 5.64 mmol.) in acetonitrile (20 mL) and IN HC1 solution (150 mL) was heated to reflux for 96 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic phases were concentrated WO 95/24200 PCT/US95/03099 88 under reduced pressure and the residue chromatographed on silica gel MeOH/chloroform/0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with anhydrous HC1. The product wa3 isolated as the dihydrochloride salt (354 mg) by filtration.

mp. 275.4 0

C.

Analysis Calculated Found C 64.28 64.21 H 6.94 7.01 N 10.71 10.74 Example 33 Preparation of 6-methyl-l-[(4-dibutylaminophenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole dihydrochloride To a stirred, cooled (-78 0 C) suspension of methoxymethyl-triphenylphosphonium chloride (8.81 g, 25.7 mmol.) in dry THF (150 mL) was added n-BuLi solution (16.1 mL. 1.6 M; 25.7 mmol.) dropwise by syringe. The orange suspension was stirred at -78 0 C for 15 min. A solution of 4-dibutylaminobenzaldehyde (5.00 g, 2.14 mmol.) in THF mL) was added to the ylide dropwise over 10 min. The reaction mixture was gradually warmed to ambient temperature and stirred 14 H. Saturated ammonium chloride solution (100 mL) was added and the mixture extracted with diethyl ether (3 X 50 mL). The combined organic phases were dried over sodium sulfate and concentrated under reduced pressure.

Chromatography on silica gel, eluting with 15% ethyl acetate/hexanes afforded product (3.47 g) as a mixture of olefin isomers which was used without further purification.

A mixture of 5-methyltryptamine hydrochloride (605 mg, 2.87 mmol.) and l-methoxy-4'-dibutylamino-styrene (1.00 g, 3.83 mmol.) in acetonitrile (20 mL) and IN HC1 solution (150 mL) was heated to reflux for 96 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with WO 95/24200 PCT/US95/03099 89 chloroform. The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel MeOH/chloroform/0.2% NH40H as eluent). The fractions containing product were pooled and conci vlrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with anhydrous HC1. The product was isolated as the dihydrochloride salt (476 mg) by filtration.

mp. 266.6 0

C.

Analysis Calculated Found C 68.05 67.92 H 8.25 8.22 N 8.82 8.74 Example 34 Preparation of 6-methyl-l-[(3-fluoro-4-methoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (0.330 g) was prepared as in Example 21 except using 3-fluoro-4-methoxybenzaldehyde (5.0 g).

A suspension of azalactone prepared above (0.30 g, 1.3 mmol.) and 5-methyltryptamine hydrochloride (0.27 g, 1.3 nmol.) in 1 N HCl (20 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform.

The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH40H as eluent). The fractions containing product were pooled and concentrated under reduced pressure.

The residue was dissolved in ethyl acetate containing 1% methanol and treated with anhydrous HC1. The product was isolated as the hydrochloride salt (170 mg) by filtration.

m/e=324.

-II-

WO 95/24200 PCT/US95/33099 Analysis Calculated Found C 66.57 66.37 H 6.15 6.16 N 7.76 Example Preparation of 6-methyl-l-[(3,4-dimethylphenyl)methyl]- 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (11.3 g) was prepared as in Example 21 except using 3,4-dimethylbenzaldehyde (25.0 g).

A suspension of azalactone prepared above (2.04 g, mmol.) and 5-methyl-tryptamine hydrochloride (2.0 g, mmol.) in 1 N HC1 (80 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient termerature and the crude product isolated by filtration. The solid was triturated with ethanol and washed with diethyl ether The product was isolated as the hydrochloride salt by filtration (1.89 m/e=304.

Analysis Calculated Found C 73.99 73.84 H 7.39 7.35 N 8.21 8.48 Example 36 Preparation of 6-methyl-l-[(2-chloro-3,4-dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole hydrochloride Azalactone (5.26g) was prepared as in Example 21 except using 2-chloro-3,4-dimethoxybenzaldehyde (10.45 g).

A suspension of azalactone prepared above (1.34 g, 4.76 mmol.) and 5-methyltryptamine hydrochloride (1.0 g, 4.75 mmol.) in 1 N HCI (30 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol and washed WO 95/24200 PCT1US95/03099 with diethyl ether. The product was isolated by filtration 1 9 g) In/e=370, mp. 244 0 C (dec.) Analysis

C

Hi

N

Calculated 61.92 5 .94 6.88 Found 61.67 5.94 6.94

I-P--

WO 95/24200 PCT/US95/03099 92 Example 37 Preparation of 6-methyl-I- [(2-chloro-3-methoxy-4hydroxyphenyl) methyl -tetrahydro-9H-pyrido indole hydrochloride Azalactone (12.4 g) was prepared as in Example 21 except using 2-chloro-3-methoxy-4-hydroxybenzaldehyde (12.0 g).

A suspension of azalactone prepared above (1.29 g, 4.82 mn' and 5-methyl-tryptamine hydrochloride (1.0 g, 4.75 mmul.) in 1 N HC1 (30 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol and washed with diethyl ether. The product was isolated by filtration (1.07 mp. 240 0 C (dec.).

Analysis Calculated Found C 61.07 60.83 H 5.64 5.71 N 7.12 7.03 Example 38 Preparation of 5-fluoro-6-methyl-- (2-chloro-3,4dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole hydrochloride A suspension of azalactone (prepared in Example 36) (2.15 g, 7.63 mmol.) and hydrochloride (prepared in Example 9) (1.0 g, 4.75 mmol.) in 1 N HC1 (80 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol and washed with diethyl ether. The product was isolated as the hydrochloride salt by filtration (1.39 m/e 388.

pg l ql3P ~A WO 95/24200 PCT/US95/03099 93 Analysis Calculated Found C 59.30 59.58 H 5.45 5.47 N 6.59 6.71 Example 39 Preparation of 6-methyl-l-(cyclohexylmethyl)-1,2,3,4tetrahydro-9H-pyrido[3,4-b] indole hydrochloride A suspension of cyclohexylacetaldehyde (631 mg mmol,) and 5-methyltryptamine hydrochloride (1.0 g, 4.3 mmol.) in ethanol (20 mL) was heated to reflux for 36 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The solid was triturated with ethanol and washed with diethyl ether. The product was isolated by filtration (731 mg). m/e=282, mp 230 0

C.

Analysis Calculated Found C 71.56 71.27 H 8.C3 8.56 N 8.78 8.64 Example Preparation of 6-methyl-1-[ (3,4-dimethoxyphenyl)-1ethyl] -12,3,4-tetrahydro-9H-pyrido[3,4-b]indole butenedioate To a stirred, cooled (-20 0 C) suspension of methoxymethyltriphenylphosphonium chloride (118.9 g, 347 mmol.) in dry THF (2000 mL) was added potassium t-butoxide (39.3 g, 350 mmol.) in portions. The orange suspension was stirred at -20 0 C for 30 min. A solution of 3,4dimethoxyacetophenone (50.0 g, 275 mmol.) in THF (500 mL) was added to the ylide dropwise over 30 min. The reaction mixture was gradually warmed to ambient temperature and stirred 2 H. Saturated ammonium chloride solution (500 mL) WO 95/24200 PCT/US95/03099 94 was added and the mixture extracted with diethyl ether (3 X 500 mL). The combined organic phases were dried over sodium sulfate and concentrated under reduced pressure.

Chromatography on silica gel, eluting with 15% ethyl acetate/hexanes afforded product (48.4 g) as a mixture of olefin isomers which was used without further purification.

A mixture of 5-methyltryptamine hydrochloride (2.16 g, 10.3 mmol.) and 1-methoxy-2-methyl-3',4'dimethoxystyrene (prepared above) (2.13 g, 10.3 mmol.) in methanol (12 mL) and 1N HC1 solution (108 mL) was heated to reflux for 96 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel MeOH/chloroform/0.2% NH40H as eluent). The fractions containing p:oduct (upper diastereomer) were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with maleic acid. The product was isolated as the maleate salt (260 mg) by filtration. mp. 187-190 0

C.

Analysis Calculated Found C 66.94 66.95 H 6.48 6.35 N 6.00 5.81 Example 41 Preparation of 6,7-dimethyl-1- [(3,4-dimethoxyphenyl) -1ethyl] 2,3,4-tetrahydro-9H-pyrido[3,4-b]indole butenedioate 5,6-dimethyltryptamine hydrochloride (prepared in Example 16) .(1.60 g, 7.12 mmol.) was converted to its free base with aqueous potassium carbonate in chloroform. This solution was dried and treated with l-methoxy-2-methyl-3',4'dimethoxystyrene (prepared above in Example 40) (1.49 g, 7.14 mmol.) and trifluororacetic acid (1.62 g, 14.2 mmol) and was WO 95/24200 PCT/US95/03099 heated to reflux for 96 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform.

The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel MeOH/chloroform/0.2% NH 4 0H as eluent). The fractions containing product (upper diastereomer) were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with maleic acid. The product was isolated as the maleate salt (560 mg) by filtration.m/e=364. mp. 177 0 C (dec.).

Analysis CalculaLed Found C 67.48 67.34 H 6.71 6.68 N 5.83 5.74 Example 42: Preparation of 6-ethyl-l- [(3,4-dimethoxyphenyl)-l-ethyll- 1,2,3,4-tetrahydro-9H-pyri3nd3,4-bindole (Z)-2-butenedioate hydrochloride (prepared in Example 17) (2.0 g, 8.9 mmol.) was converted to its free base with aqueous potassium carbonate in chloroform. This solution was dried and treated with l-methoxy-2-methyl-3',4'dimethoxystyrene (prepared above in Example 40) (1.86 g, 8.9 mmol.) and triflururacetic acid (2.03 g, 17.8 mmol) and was heated to reflux for 96 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform.

The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel MeOH/chloroform/0.2% NH40H as eluent). The fractions containing product (upper diastereomer) were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with maleic acid. The product was isolated as the maleate salt (430 mg) by filtration.m/e=364. mp. 192-194 0 C (dec.).

WO 95/24200 PCTUS95/03099 96 Analysis Calculated Found C 67.48 67.32 H 6.71 6.72 N 5.83 5.76 Example 43 Preparation of 6-methyl-l-[(3,4-dimethoxyphenyl)-lpropyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole butenedioate To methanesulfonic acid (203 mL) was added phosphorus pentoxide (30.0 g) slowly with stirring. After the addition was complete, the mixture was further stirred under nitrogen atmosphere for 2 hours until homogenous. To this solution was added 3,4-dimethoxyphenylacetonitrile g, 0.28 mol.) in a single portion, followed by 2-methyl- 2,4-pentanediol (72.1 mL, 0.56 mol.) dropwise at such a rate as to maintain a temperature between 25 and 30 0 C (1 hour).

After complete addition, the reaction mixture was stirred at ambient temperature for 10 hours and poured onto ice (500 g).

The mixture was made basic with sodium hydroxide solution added at such a rate as to keep the temperature below 0 C. The mixture was extracted with diethyl ether (3 X 250 mL) and the combined organic phases were dried over magnesium sulfate and concentrated under reduced pressure to afford a green solid. Distillation (Kugelrohr) afforded intermediate product (27.7 g) which was used without further purification.

To a stirred, cool (-78 0 C) solution of previously prepared intermediate product (27.2 g, 0.106 mol.) in THF (400 mL) under argon atmosphere was added n-butyllithium solution (68.7 mL, 1.6 M in hexanes, 0.11 mol.) dropwise via syringe over 15 minutes. After complete addition, the orange solution was stirred at -78 0 C for 30 minutes. Ethyl bromide (8.18 mL, 0.10 mol.) was added dropwise via syringe and the resulting solution further stirred at -78 0 C for 45 minutes.

n-Butyllithium (68.7 mL, 1.6 M in hexanes, 0.11 mol.) was added dropwise over '5 minutes and the orange solution i i WO 95/24200 PCT/US95/03099 97 stirred for 2 hours. The mixture was poured into ice/water (500 mL) and was acidified to pH 2-3 with 5N HC1 solution.

The mixture was extracted with diethyl ether (2 X 100 mL) and these extracts were discarded. The aqueous phase was made basic with sodium hydroxide solution cooling the mixture with ice when necessary. The basic aqueous phase was extracted with diethyl ether (2 X 200 mL) and the combined organic extracts were dried over magnesium sulfate, filtered and concentrated to afford product as an oily solid (12.08 which was used without further purification.

To a stirred cooled (-40 0 C) solution of previous product (12.0 g, 39.3 mmol.) in THF (90 mL) and ethyl alcohol mL) was added 5 N HC1 solution until pH 7. In a separate flask, a solution of sodium borohydride (2.12 g, 55.4 mmol.) was dissolved in water (20 mL) to which 1 drop of 50% sodium hydroxide had been added. Portions of the sodium borohydride solution and 5 N HCI solution were alternately added to the reaction mixture such that the pH remained 6-8, at such a rate as to maintain temperature between -35 and -45 0 C. After complete addition, the reaction mixture was warmed to ambient temperature over about 2 hours. The reaction mixture was made basic with sodium hydroxide solution and extracted with diethyl ether (3 X 100 mL). The combined organic phases were washed with brine and dried over magnesium sulfate.

Filtration and removal of solvent afforded crude product (11.3 g) as a viscous oil, which was used without further purification.

A mixture of crude product from the previous reaction (11.3 g, 36.8 mmol) and oxalic acid dihydrate (15.1 g, 120 mmol.) in water (300 mL) was heated to reflux for 12 hours. The mixture was cooled to ambient temperature and extracted with chloroform (2 X 100 mL). The combined organic phases were dried over magnesium sulfate, filtered and concentrated to afford aldehyde as an orange oil.

Distillation (Kugelrohr) under reduced pressure afforded pure aldehyde (4.97 g) as a pale oil.

WO 95/24200 PCT/US95/03099 98 A mixture of 5-methyltryptamine hydrochloride (2.53 g, 12.0 mmol.) and 2-ethyl-3',4'-dimethoxyphenylacetaldehyde (prepared above) (2.49 g, 12.0 mmol.) in mL) was heated to reflux for 48 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel MeOH/chloroform/0.2%

NH

4 0H as eluent). The fractions containing product (upper Rf diastereomer) were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with maleic acid. The product was isolated as the maleate salt (1.51 g) by filtration. m/e=364.

Analysis Calculated Found C 67.48 67.35 H 6.71 6.96 N 5.83 5.77 Example 44 Preparation of 2,6-dimethyl-l-[(2-chloro-3,4dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole hydrochloride An aqueous solution of 6-methyl-l-[(2-chloro-3,4dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido [3,4-b] indole hydrochloride (Prepared in Example 36) (500 mg, 1.23 mmol.) was treated with sodium hydroxide (49 mg, 1.23 mmol.), followed by formic acid (0.91 mL) and aqueous formaldehyde solution (0.18 mL). The mixture was heated to reflux for 4 H. The reaction mixture was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between aqueous potass.um carbonate solution and diethyl ether. The organic phase was dried over potassium carbonate and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with maleic acid. The maleate salt was isolated by filtration and WO 95/24200 PCT/US95/03099 99 purified by recrystallization from ethyl acetate/hexanes to afford product (240 mg). m/e=385.

Analysis Calculated Found C 62.34 62.47 H 5.84 5.71 N 5.59 5.58 Example Preparation of 2-methyl-6-(l-methylethyl)-1,2,3,4-tetrahydro- 1- (3,4-dimethoxyphenyl)methyl] -9H-pyrido- [3,4b] indole maleate An aqueous solution of 6-(l-methylethyl)-l-[(3,4dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole (Z)-2-butenedioate (prepared in Example 13) (500 mg, 1.04 mmol.) was treated with sodium hydroxide (83 mg, 2.08 mmol.), followed by formic acid (0.77 mL) and aqueous formaldehyde solution (0.15 mL). The mixture was heated to reflux for 4 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic layers were concentrated under reduced pressure and the residue chromatographed on silica gel (ethyl acetate 0.2% NH40H as eluent). The fractions containing product weire pooled and concentrated under reduced pressure.

The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (130 mg) by filtration. m/e=376.

Analysis Calculated Found C 67.99 67.88 H 6.93 6.73 N 5.66 5.69 WO 95/24200 PCT/US95/03099 100 Example 46 Preparation of (-)-(S)-6-methyl-1,2,3,4-tetrahydro-l-[(3,4dimethylphenyl)methyl]-9H-pyrido[3,4-b]indole hydrochloride To a stirred solution of 6-methyl-l,2,3,4tetrahydr 9H-pyrido[3,4-b]indole (3.14 g, 16.9 mmol.) in dry xylenes (65 rnL) was added (S)-N,N-dimethyl-N'-(1-cert-butoxy- 3-methyl)-2-butylformamidine (3.79 g 17.7 mmol.) followed by camphorsulfonic acid (200 mg). ThB resulting solution was heated to reflux for 72 hours. The solution was cooled to ambient temperature and concentrated under reduced pressure.

The residue was purified by flash chromatography on silica gel (1:3:6 triethylamine:ethyl acetate:hexanes as eluent).

The product containing fractions were pooled and concentrated to afford the product formamidine (5.99 g) as a viscous oil which was used without further purification.

To a stirred, cooled suspension of potassium hydride (25% oil dispersion, 829 mg, 20.2 mmol.) in THF mL) was added formamidine prepared above (5.99 g 16.8 mmol.) in THF (45 mL). To this mixture was added tetramethylethylenediamine (3.0 mL, 20.2 mmol.) followed by chloromethylmethyl ether (1.9 mL, 25.2 mmol.). The mixture was stirred for an additional 1 hour and treated with water mL). The mixture was partitioned between diethyl ether and water and the layers separated. The aqueous phase was extracted with diethyl ether (2 X 100 mL) and the organic phases combined, dried over potassium carbonate, and concentrated to afford product (6.73 g) as an orange oil, which was used without further purification.

To a stirred, cooled (-78 0 C) solution of previously prepared formamidine (6.29 g, 8.4 mmol.) in dry THF (100 mL) was added n-BuLi (1.7 M solution in hexanes, 10.1 mL, 17.1 mmol.) dropwise over 5 minutes. The solution was further stirred at -78 0 C solution for 1 hour and treated with 1chloromethyl-3,4-dimetho;-cbenzene (3.35 g, 17.9 mmol.) in dry THF (15 mL). The solution was further stirred for 4 hours at -78 0 C and allowed to warm to roonm temperature overnight. Wet THF was added (50 mL) and the solution was concentrated under WO 95/24200 PCT/US95/03099 101 reduced pressure. The residue was dissolved in chloroform and washed with water. The organic phase das dried over sodium carbonate and concentrated. The crude product was purified by flash chromatography on silica gel (1:3:6 triethylamine:ethyl acetate: hexanes as eluent). The product containing fractions (upper Rf) were pooled and concentrated to afford 'roduct (3.92 g) as a viscous oil (m/e 55 which was used without further purification.

To a stirred solution of methoxymethylindole prepared above (3.92 g 7.13 mmol.) in THF (70 mL) was added 2N HC1 (20 mL). The mixture was stirred at ambient temperature for 24 hours, and partitioned between diethyl ether and water. The aqueous phase was back extracted with diethyl ether (2 X 50 mL) and the combined organic phases were washed with brine, dried over sodium carbonate, and concentrated under reduced pressure. The residue was dissolved in THF (20 mL) and treated with 2N sodium hydroxide solution (6 mL). After 2 hours, the reaction mixture was extracted with chloroform (2 X 100 mL). The organic phase was dried over sodium carbonate and concentrated.

Chromatography on silica gel (1:3:6 triethylamie/ethyl acetate/hexanes as eluent) afforded product (1.85 g) as a viscous oil 505).

To a stirred, cooled solution of previously prepared formamidine (1.37 g, 5.41 mmol.) in ethanol (50 mL) was added water (6 mL) followed by acetic acid (6 mL) and hydrazine hydrate (11 mL). The reaction vessel was placed in the freezer (-10 0 C) for 72 hours. The mixture was warmed to ambient temperature and concentrated under reduced pressure.

The crude product was dissolved in chloroform (300 mL) and washed with water (3 X 50 mL). The organic phase was dried over sodium carbonate and concentrated to a viscous oil. The oil was dissolved in diethyl ether and treated with anhydrous HC1. The hydrochloride salt (560 mg) was isolated by filtration. Recrystallization from ethanol (2 X) afforded material of constant rotation. Chiral HPLC confirmed enantiomeric purity as 98% ee. m/e=336) WO 95/24200 WO 9524200PCT[US95/03099 102 specific rotation 589 nN -118.0 (pyridine, 0=1) specific rotation 365 nM -401.0 (pyridine, C=1) Analysis

C

H

N

Calculated 67 .64 7.51 Found 67 6.70 7.52 ExgMl1e 47 Preparation of 6-methyl-1-(1-(4-methoxynaphthaleny1)mv-thyl) 3,4-tetrahydro-9H-pyrido indole hydroi-hloride

CHO

OMe

CO

2 Ac2O, NaOAc, 100 0

C

OMEN

A solution of 4-methoxy-l-naphthaldehyde (20.0 g, 0.107 mol. N-acetylglycine 12.58 g, 0.107 mol. )and sodium acetate 8.81 g, 0.107 mol in acetic anhydride 100 mL was heated to 1000C for 2 hours, The reaction mixture was cooled to ambient temperature and stirred for hours jander nitrogen atmosphere. The mixture was poured onto ice 2520 irL with stirring. The product was isolated by filtration, washed with water 3 X 50 InL and diethyl ether (3 X 50 mL and dried under reduced pressure 3.16 g WO 95/24200 WO 9524200PCT/US95/03099 103 Ae supnio of azalctoe prae d above (1.1 IN 5.62 io.)iiNHl(0L ws He orfu for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The brown solid was triturated with isopropyl alcohol (3 X 50 mL) and washed with diethyl ether (3 X 50 inL) Recrystallization from ethanol afforded 1.42 g of desired product as a pale solid. (mp 271.7 0

C)

An~ilsi Fon~ C 73.36 73.60 H 6.41 6.51 N 7.13 7.20 Bxmpnle-48 Preparation of 6 -methyl-i- (1-(2-methoxy-.-,ipthaelyl) methyl)1,2,3,4-tetrahydro-9H-pyrido[3,4-bJ- indole hydrochloride To a stirred, cooled (-78 0 C) solution of miethc. xymethyl-triphenylphosPhoniufl chloride (11.05 g, 32.2 nmol.) in 150 mL of anhydrous THF was added n-butyllithium (20.14 mL of 1.6 M solution in hexanes, 32.2 mmol) dropwise via syringe. After complete addition, the solution was stirred at this temperature f~or 15 min. A solution of 2methoxy-l-narC.hthaldehyde (5.0 g, 26.9 iilmol.) in THF (75 mL) WO 95/24200 PCT/US95/e .099 104 was added to the solution dropwise by addition funnel. After complete addition, the solution was allowed to warm to ambient temperature and stir for 14 hours. A saturated solution of ammonium chloride (100 mL) was added and the mixture was partitioned between diethyl ether and water. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by plug filtration (silica gel, eluent ethyl acetate/hexanes) and afforded 5.0 g of product as a .mixture of enol ethers, which was used without further purification.

A solution of enol ethers prepared above (5.0 g, 23.3 mmol.) in diethyl ether (50 mL) was treated with water mL) and perchloric acid (1.5 mL of 60% solution). The solution was stirred at ambient temperature for 72 hours.

The solution was diluted with chloroform (100 mL) and neutralized with saturated sodium bicarbonate solution. The mixture was extracted with chloroform (3 X 100 mL and the combined organic phases were dried over sodium sulfate and concentrated. The residue was purified by flash chromatography on silica gel diethyl ether/hexanes as eluent) to afford (2-mc-hoxy-l-naphthyl)-acetaldehyde (1.79 g) as a colorless oil.

To a stirred solution of hydrochloride (947 mg, 4.49 mmol.) in 20 mL of ethyl alcohol was added (2-methoxy-l-naphthyl)-acetaldehyde (l.0g, 4.99 mmol.). The solution was heated to reflux under nitrogen atmosphere for 40 hours. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. Recrystallization from ethyl alcohol/2butanone afforded product as a pale solid (705 mg). (mp. 245.3 0

C).

Analysis: Calculated gound C 73.36 73.29 H 6.41 6.64 WO 95/24200 PCT/US95/03099 105 N 7.13 7.12 Example 49 Preparation of 6-methyl-l-(1-naphthalenyl-1-ethyl)- 1,2,3,4-tetrahydro-9H-pyrido indole 2butenedioate Me CN OH OH -Me Me- Me P205, MeSO 3

H

To methanesulfonic acid (215 mL) was added phosphorus pentoxide (31.8 g) slowly with stirring. After the additibn was complete, the mixture was further stirred under nitrogen atmosphere for 2 hours until homogeneous. To this solution was added 1-naphthyl acetonitrile (50 g, 0.3 mol.) in a single portion, followed by 2-methyl-2,4pentanediol (76.4 mL, 0.6 mol.) dropwise at such a rate as to maintain a temperature between 25 and 30 0 C (1 hour). After complete addition, the reaction mixture was stirred at ambient temperature for 10 hours and poured onto ice (500 The mixture was made basic with sodium hydroxide solution added at such a rate as to keep the temperature below 35 0 C. The mixture was extracted with diethyl ether (3 X 250 mL) and the combined organic phases were dried over magnesium sulfate and concentrated under reduced pressure to afford a green solid. Recrystallization from ethyl acetate afforded product (28.29 g which was used without further purification.

WO 95/24200 PCT/US95/03099 106 Me Me 0 0 Me Me Me N

N

Me Me t-BuLi,THF,-7 C; I t-BuLi To a stirred, cooled (-78 0 C) solution of previously prepared "isoxazan?" (28.3 g, 0.106 mol.) in THF (475 mL) under argon atmosphere was added t-butyllithium solution (68.4 mL, 1.7 M in pentane, 0.116 mol.) dropwise via syringe over 15 minutes. After complete addition, the orange solution was stirred at -78 0 C for 30 minutes. Methyl iodide (6.6 mL, 0.106 mol.) was added dropwise via syringe and the resulting solution further stirred at -78 0 C for 45 minutes.

t-Butyllithium (68.4 mL, 1.7 M in penta.ae, 0.116 mol.) was added dropwise over 15 minutes and the orange solution stirred for 2 hours. The mixture was poured into ice/water (500 mL) and was acidified to pH 2-3 with 5N HC1 solution.

The mixture was extracted with diethyl ether (2 X 100 mL) and these extracts were discarded. The aqueous phase was made basic with sodium hydroxide solution cooling the mixture with ice when necessary. The basic aqueous phase was extracted with diethyl ether (2 X 200 mL) and the combined organic extracts were dried over magnesium sulfate, filtered and concentrated to afford product as an oily solid (13.15 which was used without further purification.

WO 95/24200 PCTIUS95/03099 107 Me Me eMe 0 0 Me Me Me Me Me N NaBH 4 ,THF,EtOH H Me To a stirred cooled (-40 0 C) solution of previous product (13.15 g, 46.7 mmol.) in THF (100 mL) and ethyl alcohol (100 mL) was added 5N HC1 solution until pH 7. In a separate flask, a solution of sodium borohydride (2.52 g, 65.8 mmol.) was dissolved in water (20 mL) to which 1 drop of sodium hydroxide had been added. Portions of the sodium borohydride solution and 5 N HC1 solution were alternately added to the reaction mixture such that the pH remained 6-8, at such a rate as to maintain temperature between -35 and 0 C. After complete addition, the reaction mixture was warmed to ambient temperature over about 2 hours. The reaction mixture was made basic with sodium hydroxide solution and extracted with diethyl ether (3 X 100 mL). The combined organic phases were washed with brine and dried over magnesium sulfate. Filtration and removal of solvent afforded crude product (13.2 g) as a viscous oil, which was used without further purification.

Me 0 Me Me Me CHO

N

H Me oxalic acid, H C N' WO 95/24200 PCT/US95/03099 108 A mixture of crude product from the previous reaction (13.2 g, 46.6 mmol) and oxalic acid dihydrate (19.1 g, 152 mmol.) in water (380 mL) was heated to reflux for 12 hours. The mixture was cooled to ambient temperature and extracted with chloroform (2 X 100 mL). The combined organic phases were dried over magnesium sulfate, filtered and concentrated to afford aldehyde as an orange oil.

Distillation (Kugelrohr) under reduced pressure afforded pure aldehyde (1.97 g) as a pale oil.

Me NH maleate Me CHO Me Me NH2 HCI EtOH, reflux

K

2

CO

3 c" f r ace m i c "N i maleic acid A

H

Me r NHmaleate Me N H racemic |i A solution of 5-methyl tryptamine hydrochloride (1.11 g, 5.27 mmol.) and 2-(1-naphthyl )-propionaldehyde (0.97 g, 5.26 mmol.) in 95% ethyl alcohol was heated to reflux for 48 hours under nitrogen atmosphere. The mixture was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between aqueous potassium carbonate solution and chloroform. The chloroform phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was subjected to flash chromatography on silica gel (25% methyl alcohol in chloroform as eluent), affording 529 mg of the higher rf isomer and 200 mg of the lower rf isomer. Each diastereomer, independently, was dissolved in ethyl acetate and treated with excess maleic acid. The maleate salts were WO 95/24200 PCTIUS95/03099 109 isolated by filtration affording 570 mg of isomer A and 30 mg of isomer B.

isomer A data: m/e 340 Analysis Calculated Found C 73.66 73.64 H 6.18 6.13 N 6.14 6.44 isomer B data: m/e 340 Analysis Calculated Found C 73.66 73.41 H 6.18 6.04 N 6.14 5.89 Example Preparation of 6-(l1,-dimethyhlethyl)--(l-naphthalenyl- 1-ethyl)-1,2,3,4-tetrahydro-9H-pyrido indole hydrochloride 4-Chlorobutyryl chloride (300 g, 2.13 mol.) was dissolved in dry THF (3 L) To this solution was added 2,6lutidine (252 mL) followed by 5% Pd/C (30 This mixture was placed in a Parr hydrogenator and shaken under 60 psi of hydrogen for 6 hours. The mixture was purged with nitrogen, filtered, washing the catalyst with THF (500 mL), and concentrated at room temperature under reduced pressure. Distillation afforded 4-chlorobutanal (148.3 g) as a colorless liquid.

To a stirred suspension of 4-isopropylphenylhydrazine hydrochloride monohydrate (15.3 g, 91.95 mmol.) in chloroform (250 mL) was added saturated sodium carbonate solution (250 mL). The mixture was stirred for 30 minutes until the organic phase appeared homogeneous, WO 95/24200 PCT/US95/03099 110 and extracted with chloroform (2X 200 mL). The combined organic phases were concentrated to afford the hydrazine free base as a yellow oil. This oil was dissolved in methanol (200 mL) and water (5 mL) and treated with sodium acetate (6.72 g, 82 mmol.) and 4-chlorobutanal (8.7 g, 82 mmol.).

The mixture was.placed in a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oil bath preheated to 100 0 C. Heating was continued for 18 hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were combined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HC1 gas. The hydrochloride salt was isolated -v filtration, washed with 2-propanol mL) and diethyl ether (100 mL) and dried to afford isopropyl-tryptamine hydrochloride (9.8 g) as a pale solid, which was used without further purification.

A solution of 5-isopropyltryptamine hydrochloride (1.24 g, 5.19 mmol.) and 2-(1-naphthyl)-propionaldehyde (0.95 g, 5.16 mmol.) in 95% ethyl alcohol was heated to reflux for 48 hours under nitrogen atmosphere. The mixture was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between aqueous potassium carbonate solution and chloroform. The chloroform phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was subjected tc flash chromatography on silica gel (25% methyl alcohol in chloroform as eluent), affording 500 mg of the higher rf isomer along with 400 mg of impure lower r2 isomer. The major diastereomer was dissolved in ethyl acetate and treated with excess maleic acid. The maleate salt was isolated by filtration affording 400 mg of named product as a pale solid.

WO 95/24200 PCT/US95103099 111 m/e=369.

Analysis Calculated Found C 74.36 74.58 H 6.66 6.64 N 5.78 5.81 Example 51 Preparation of 6-methyl-- (1-naphthalenylmethyl) 1,2,3,4-tetrahydro-9H-pyrido indole hydrochloride A solution of l-naphthaldehyde (25.0 g, 0.16 mol.), N-acetylglycine (19.0 g, 0.162 mol.) and sodium acetate (13.1 g, 0.160 mol) in acetic anhydride (147 mL) was heated to 100 0 C for 4 hours. The reaction mixture was cooled to ambient temperature poured onto ice (300 mL) with stirring.

The product was isolated by filtration, washed with water (3 X 50 mL) and diethyl ether (3 X 50 mL) and dried under reduced pressure (11.82 g).

A suspension of azalactone prepared above (3.15 g, 13.3 mmol.) and 5-methyl-tryptamine hydrochloride g, 9.5 mmol.) in IN HC1 (50 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The brown solid was triturated with isopropyl alcohol (3 X 50 mL) and washed with diethyl ether (3 X mL). Recrystallization from -I 1- WO 95/24200 PCT/US95/03099 112 ethanol afforded 1.94 g of desired product as a pale solid.

Analysis Calculated Found C 76.12 76.03 H 6.39 6.22 N 7.72 7.52 Example 52 Preparation of 8-bromo-l-(1-naphthalenylmethyl)- 1,2,3,4-tetrahydro-9H-pyrido indole hydrochloride NH2

HCI

NIHNH

2

K

2

CO

3 ,CHC13; MeOH,95 0

C;

Br Et 2 0, MeOH, iCl

HN

CI HBr 0 O To a stirred suspension of 2-bromophenyl-hydrazine hydrochloride (25.8 g, 115 mmol.) in chloroform (500 mL) was added saturated sodium carbonate solution (500 mL). The mixture was stirred for 30 minutes until the organic phase appeared homogenous, and extracted with chloroform (2 X 200 mL). The combined organic phases were concentrated to afford the hydrazine free base as a yellow oil. This oil was dissolved in methanol (100 mL) and treated slowly with 4-chlorobutanal (prepared a- described in Example 4) (12.3 g, 115 mmol.). The mixture was placed in a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oil bath preheated to 95 0 C. Heating was continued for 18 hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium WO 95/24200 PCT/US95/03099 113 carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were combined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HC1 gas. The hydrochloride salt was isolated by filtration, washed with 2-propanol mL) and diethyl ether (100 mL) and dried to afford 7bromo-tryptamine hydrochloride (3.6 g) as a pale solid, which was used without further purification.

A suspension of azalactone (prepared as described in Example 5) (55 g, 6.53 mmol.) and 7-bromo-tryptamine hydrochloride (1.50 g, 5.44 mmol.) in IN HC1 (100 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The brown solid was triturated with isopropyl alcohol (3 X 50 mL) and washed with diethyl ether (3 X 50 mL).

Recrystallization from ethanol afforded 260 mg of desired product as a pale solid. (mp 231-2330C, dec.) Analys is Calculated Found C 61.77 61.48 H 4.71 4.63 N 6.55 6.73 Example 53 Preparation of 8-bromo-l-(l-naphthalenylmethyl)- 1,2,3,4-tetrahydro-9H-pyrido indole hydrochloride To a stirred suspension of 2-bromophenyl-hydrazine hydrochloride .(25.8 g, 115 mmol.) in chloroform (500 mL) was added saturated sodium carbonate solution (500 mL). The mixture was stirred for 30 minutes until the organic phase appeared homogenous, and extracted with chloroform (2 X 200 mL). The combined organic phases were WO 95/24200 PCT/US95/03099 114 concentrated to afford the hydrazine free base as a yellow oil. This oil was dissolved in methanol (100 mL) and treated slowly with 4-chlorobutanal (prepared as described in Example 4) (12.3 g, 115 mmol.). The mixture was placed in a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oil bath preheated to 95 0 C. Heating was continued for 18 hours. The resulting dark solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between chloroform/methanol (75/25 by volume) and aqueous sodium carbonate solution. The organic phase was concentrated and the crude indole ethanamine was purified by flash chromatography on silica gel (0-25% methanol gradient in chloroform as eluent). Fractions containing product were combined and concentrated. The oil was dissolved in diethyl ether (300 mL) containing 1% methanol and treated with dry HC1 gas. The hydrochloride salt was isolated by filtration, washed with 2-propanol mL) and diethyl ether (100 mL) and dried to afford 7bromo-tryptamine hydrochloride (3.6 g) as a pale solid, which was used without further purification.

A suspension of azalactone (prepared as described in Example 5) (55 g, 6.53 mmol.) and 7-bromo-tryptamine hydrochloride (1.50 g, 5.44 mmol.) in IN HC1 (100 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration. The brown solid was triturated with isopropyl alcohol (3 X 50 ni) and washed with diethyl ether (3 X 50 mL).

Recrystallization from ethanol afforded 260 mg of desired product as a pale solid. (mp 231-2330C, dec.) Analysis Calculated Found C 61.77 61.48 H 4.71 4.63 N 6.55 6.73 WO 95/24200 PCT/US95/03099 115 ExamPple 54 Preparation of 8-methoxy-l-(1-naphthalenylmethyl)- 1,2,3,4-tetrahydro-9H-pyrido indole 3)-2butenedioate To a stirred, cooled suspension of 2methoxyphenylhydrazine hydrochloride (14.44 g, 83 mmol.) in THF (600 mL) was added 4-chlorobutanal prepared as described in Example 5 (9.0 g, 84 mmol.) followed by dropwise addition of triethylamine (8.6 g, 85 mmol.) in THF (20 mL) Upon complete addition, the cooling bath was removed and the solution stirred for 1 hour. The reaction mixture was filtered and the filter cake washed with THF (100 mL). The combined filtrates were concentrated to an orange oil, which was dissolved in methanol (150 mL) and water (5 mL). Th solution was transferred to a sealable tube and purged with nitrogen for 10 minutes. The tube was sealed and placed in an oilbath preheated to 95 0 C. After heating for 14 hours, the reaction mixture was cooled to ambient temperature and concentrated under reduced pressure. The residue was partitioned between saturated aqueous potassium carbonate and 3:1 chloroform: 2-propanol. The organic phase was dried over sodium sulfate and concentrated. The residue was purified by flash chromatography on silica gel (15% methanol, 0.2% in chloroform as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in methanol and treated with dry HC1 and concentrated to afford 7-methoxytryptamine hydrochloride (4.04 g) as a stable foam, which was used without further purification.

A suspension of azalactone (preparedc as described in Example 5) (1.30 g, 5.5 mmol.) and 7-methox/tryptamine hydrochloride (1.08 g, 4.8 mmol.) in IN HCI (100 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was 9 WO 95/24200 PCT/US95/03099 116 removed under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH40H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (880 mg by filtration.

(mp 226-227 0 C, dec.) Analysis Calculated Found C 70.73 70.61 H 5.72 5.77 N 6.11 6.03 Example Preparation of 6-bromo-l-(1-naphthalenylmethyl)- 1,2,3,4-tetrahydro-9H-pyrido indole butenedioate "A suspension of azalactone (prepared as described in Example 5) (1.4 g, 5.9 mmol.) and hydrochloride (1.77 g, 6.4 mmol.) in IN HC1 (100 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was removed under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (540 mg) by filtration.

(mp 234-235 0 C, dec.) m/e= 390.

-O MOMEMOOMW WO 95/24200 PCT/US95/03099 117 Analysis Calculated Found C 61.55 61.38 H 4.57 4.64 N 5.52 5.29 Example 56 Preparation of 7-methyl-8-bromo-l-(1-naphthalenyl methyl)-1,2,3,4-tetrahydro-9H-pyrido indole hydrochloride 2-.Bromo-3-methylphenylhydrazine hydrochloride (23 g) was prepared as described for 2-bromo-4 methylphenylhydrazine hydrochloride in Example 7, except using 2-bromo-3-methylaniline as starting material.

O ,I

SNHNH

2

K

2

CO

3 CHC13; H 2

HCI

Me MeOH,95 0

C;

Br Et 2 0,MeOH,HCl Me

N

0 Br 6-Methyl-7-bromotryptamine hydrochloride was prepared (2.42 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 7, except using 2-bromo- 3-methyl phenylhydrazine hydrochloride as starting material.

A suspension of azalactone (prepared as described in Example 5) (1.07 g, 4.51 mmol.) and 6-methyl-7bromotryptamine hydrochloride (1.22 g, 4.21 mmol.) in IN HC1 mL) was heated to reflux for 65 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was removed under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH40H as eluent). The fractions containing product were pooled and ~u -I1I WO 95/24200 WO 9524200PCT/US9S/03099 118 concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with dry HC1. The product was isolated as the hydrochloride salt (840 mg) by filtration. (mLp 276-279C.. dec.) Analysis Calculated Found 62 .7 9 4.96 62 .53 5.02 6.

Exam Preparation of 6-cyclohe 1,2,3, 4-tetrahydro-9H-pyrido 4-Cyclohexyl phenyli (35.6 g) was prepared as descr phenylhydrazine hydrochloride cyclohexyl-aniline as starting '''NHNH,

K

2 co 31 dllc MeOH, 95 0

C

Et 2 O MeOH

H

0 34 6.19 57 ~xYi-1- (I-neaphthalenylmethyl) indole hydrochloride Lydrazine hydrochloride ~ibed for 2-bromo-4 met'hylin Example 7, except using 4material.

H

ne hydrochloride was prepared thyl-7 -bromotryptaroine cept using 4ochloride as starting material.

actone (prepared as describeI ol.) and ol.) in I N HCl (70 mL) was under nitrogen atmosphere. The ohexyltryptami (1.29 g) as described for i,-me hydrochloride in Example 7, ex cyclohexylphenylhydrazine hydr A suspension of azal in Example 5) (1.09 g, 4.59 mm hydrochloride (1.28 g, 4.59 mm heated to ref Thx 14 hours WO 95/24200 PCT/US95/03099 119 reaction mixture was cooled to ambient tcnperature and the crude product isolated by filtration. The solid was recrystallized from ethanol (2X) to afford 690 mg of desired product as a pale solid hydrochloride salt. m/e= 395 Analysis Calculated Found C 78.03 78.26 H 7.25 7.06 N 6.50 '48 Example 58 Preparation of 2,6-dimethyl-l-(1-naphthalenylmethyl)- 1, .3,4-tetrahydro-9H-pyrido indow1, hydrochloride To an aqueous solution (200 mL) of 5-methyl-l-(1naphthalenylmethyl)-l,2,3,4-tetrahydro-9H-pyrido [3,4-b] indole hydrochloride (2.00 g, 5.51 mmol.) previously prepared in Example 5, was added formic acid (4.1 mL) and formaldehyde solution (0.8 mL of 37% aqueous solution). The mixture was heated to reflux for 72 hours. The solution was made basic with saturated potassium carbonate solution and extracted with chloroform (2 X 100 mL). The combined organic phases were dried over potassium carbonate and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel (chloroform as eluent) The fractions containing product were pooled and concentrated to a viscous oil. The oil was dissolved in diethyl ether and treated with anhydrous HC1 and the resulting hydrochloride salt was isolated by filtration. Drying afforded the named product (1.34 m/e P 340.

Analyis Calculated Found C 76.48 76.58 F 5.68 6.63 N 7.43 7.28 I WO 95/24200 PCT/US95/03099 120 Example 59 Preparat on of 5-fluoro-6-methyl-l-(l-naphthalenylmethyl)-1,2,3,4-tetrahydro-9H-pyrido indole butenedioate 3-fluoro-4-methyl-phenylhydrazine hydrochloride (21.4 g was prepared as described for 2-bromo-4 methylphenylhydrazine hydrochloride in Example 7, except using 3fluoro-4-methyl aniline as starting material.

F

Me NHNH K 2

CO

3 CHC1 3 NH2 Hl MeOH,95 0 C; Me Et 2 O,MeOH,

HCI

H

N

CI H s H hydrochloride was prepared (2'.20 g) as described for 5-methyl-7-bromocryptamine hydrochloride in Example 7, except using 3-fluoro- 4-methyl-phenylhydrazine hydrochloride (6.00 g) as starting material.

A suspension of azalactone (prepared as described in Example 5) (2.3 g, 9.6 mmol.) and methyltryptamine hydrochloride (2.2 g, 9.6 mmol.) in 1N HC1 mL was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was remove. under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH40H as eluent). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the i i~Lq iii WO 95/24200 PCT/US95/03099 121 maleate salt (520 mg) by filtration.

Analysis Calculated Found C 70.42 70.45 H 5.47 5.41 N 6.08 6.10 Example Preparation of 7,8,9,10-tetrahydro-10-(1naphthalenylmethyl)-11H-benzo pyrido indole 2-butenedioate

NHNH

NHNH

2

K

2

C

3 ,CHC13; NH HCI 0

C;

6+ Et 2 0,MeOH,HCl C^l

HN

0 6,7-benzotryptamine hydrochloride was prepared (2.85 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 7, except using 1-naphthyl-hydrazine hydrochloride (6.00 g) as starting material.

A suspension of azalactone (prepared as described in Example 5) (2.75 g, 11.6 mmol.) and 6,7-benzotryptamine hydrochloride (2.85 g, 11.6 mmol.) in IN HCI (50 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was removed under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH40H as eluent). The fractions containing product were pooled and I- WO 95/24200 PCT/US95/03099 122 concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (300 mg) by filtration.

m/e 363.

Analysis Calculated Found C 75.30 75.04 H 5.48 5.36 N 5.85 5.76 Example 61 Preparation of 6-(1,1-dimethylethyl)-1,2,3,4tetrahydro-- (l-naphthalenylmethyl)-9H-pyrido [3,4b] indole (Z)-2-butenedioate t-Bu St-Bu NHa 2

HCI

NHNH

2 HCI K 2 C03, CHC1 3 r E2 H EtM20,eOH,HC N/

CI

H

O

5-(1,1-dimethylethyl)-tryptamine hydrochloride was prepared (2.95 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 7, except using 4-(1,1dimethylethyl)-phenyl hydrazine hydrochloride (6.00 g) as starting material.

A suspension of azalactone (prepared as described in Example 5) (1.25 g, 5.26 mmol.) and 5-(1,1-dimethylethyl)tryptamine hydrochloride (1.33 g, 5.26 mmol.) in 1N HCl mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was removed under reduced pressure and the residue

-T,

WO 95/24200 PCT/US95/03099 123 chromatographed on silica gel (ethyl acetate/0.2% NH 4 0H as eluert). The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate containing 1% methanol and treated with maleic acid. The product was isolated as the maleate salt (700 mg) by filtration. m/e=369 Analvys Calculated Found C 74.36 74.08 H 6.66 6.69 N 5.78 5.69 EmpLe 62 Preparation of 6- (-methylethyl)-l,2,3,4-tetrahydro-l- (1-naphthalenylmethyl)-9H-pyrido [3,4b] indole butenedioate A suspension of azalactone (prepared as described in Example 5) (1.75 g, 7.38 mmol.) and hydrochloride (prepared as described in Example 4) (1.76 g, 7.37 mmol.) in 1N HC1 (40 mL) was heated to reflux for 24 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The solvent was removed under reduced pressure and the residue chromatographed on silica gel (ethyl acetate/0.2% NH 4 0H as eluent). The fractions containing product were pooled and concentrated under reduced pressure.

The residue was dissolved in ethyl acetate containing 1% methano.

1 and treated with maleic acid. The product was isolated as the maleate salt 671 mg by filtration.

m/e=355 Analysis Calculated Found C 74.02 74.08 H 6.43 6.21 N 5.95 5.83 WO 95/24200 PCTUS95103099 124 Example 63 Preparation of 6,9-dimethyl-1,2,3,4-tetrahydro-l-(1naphthalenylmethyl)-9H-pyrido indole hydrochloride To a stirred suspension of hydrochloride (10.0 g, 43.2 mmol.) in chloroform (300 mL) was added saturated sodium carbonate solution (300 mL). The mixture was stirred at ambient temperature for 1 hour. The layers were separated and the aqueous layer was back extracted with chloroform (2 X 100 mL). The combined organic layers were dried over sodium sulfate and concentrated. The residue was dissolved in toluene (300 mL) and treated with phthalic anhydride (7.05 g, 47.6 mmol.). The solution was heated to reflux for 14 hours with azeotropic removal of water (by Dean-Stark trap). The solution was cooled to ambient temperature and concentrated to afford crude product as a pale foam. Recrystallization from ethanol afforded product phthalimide (13.52 g) as a white solid, which was used without further purification.

To a stirred, cooled suspension of potassium hydride (25% oil dispersion, 8.24 g, 51.3 mmol.) in dry THF mL) was added a solution of phthalimide prepared above (13.02 g, 42.8 mmol.) in THF (150 mL) over 30 minutes. After complete addition, the mixture was further stirred for 1 hour. Tetramethyletbylenediamine (7.7 mL 51.3 mmol.) was added, followed by methyl iodide (4.0 mL, 63.8 mmol.). After 1 hour, the reaction was quenched by addition of water (200 mL), followed by extraction with diethyl ether (2 X 100 mL).

The combined organic phases were dried over magnesium sulfate and concentrated under reduced pressure to afford product as a yellow solid (14 g) which was used without further purification.

A solution of phalimide prepared in the previous step (14 g, 42.8 mmol.) in methanol (85 mL) was treated with hydrazine (3.4 mL, 109 mmol.). The mixture was heated to reflux for 2 hours. The mixture was cooled to ambient temperature, treated with concentrated HC1 (7 mL) and I i; WO 95/24200 PCT/US95/03099 125 methanol (25 mL), and further heated to reflux for 14 hours.

After cooling to ambient temperature, the mixture was partitioned between chloroform (200 mL) and saturated aqueous sodium carbonate solution (200 mL). The aqueous layer was further extracted with chloroform (2 X 100 mL) and the organic phases combined, dried over sodium sulfate and concentrated. The crude product was purified by flash chromatography on silica gel (0-25% methanol in chloroform/0.2% NH 4 0H as eluent). The product containing fractions were pooled and concentrated under reduced pressure. The residue was dissolved in diethyl ether and treated with anhydrous HC1. The product tryptamine hydrochloride (6.08 g) was isolated by filtration as a tan solid and used without further purification.

A suspension of azalactone prepared as described in Example 5 (1.06 g, 4.45 mmol.) and 1,5-dimtn hyl-tryptamine hydrochloride (1.00 g, 4.47 mmol.) in IN HC1 mL) was heated to reflux for 48 hours under nitrogen atmosphere. The reaction mixture was cooled to ambient temperature and the crude product isolated by filtration.

The brown solid was triturated with isopropyl alcohol (3 X mL) and washed with diethyl ether (3 X 50 mL).

Recrystallization from ethanol afforded 710 mg of desired product as a pale solid.

m/e=340.

Analysis Calculated Found C 76.48 76.78 H 6.68 6.58 N 7.43 7.50 Example 64 Preparation of (-)-(S)-6-methyl-1,2,3,4-tetrahydro-l-(1naphthalenylmethyl)-9H-pyrido indole hydrochloride To a stirred solution of 6-methyl-l,2,3,4tetrahydro-9H-pyrido [3,4-b]indole (3.14 g, 16.9 mmol.) in I WO 95/24200 PCT/US95/03099 126 dry xylenes 65 mL was added (S)-N,N-dimethyl-N'-(1-tertbutoxy-3-methyl)-2-butylformamidine (3.79 g 17.7 mmol.) followed by camphor sulfonic acid (200 mg). The resulting solution was heated to reflux for 72 hours. The solution was cooled to ambient temperature and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (1:3:6 triethylamine:ethyl acetate:hexanes as eluent). The product containing fractions were pooled and concentrated to afford the product formamidine (5.99 g) as a viscous oil which was used without further purification.

To a stirred, cooled suspension of potassium hydride (25% oil dispersion, 829 mg, 20.2 mmol.) in THF mL) was added formamidine prepared above (5.99 g, 16.8 mmol.) in THF (45 mL). To this mixture was added tetramethylethylenediamine (3.0 mL, 20.2 mmol.) followed by chloromethylmethyl ether (1.9 mL, 25.2 mmol.).

The mixture was stirred for an additional 1 hour and treated with water (50 mL). The mixture was partitioned between diethyl ether and water and the layers separated. The aqueous phase was extracted with diethyl ether (2 X 100 mL) and the organic phases combined, dried over potassium carbonate, and concentrated to afford product (6.73 g) as an orange oil, which was used without further purification.

To a stirred, cooled (-78 0 C) solution of previously prepared formamidine (3.36 g, 8.4 mmol.) in dry THF (55 mL) was added n-BuLi (1.7M solution in hexanes, 5.4 mL, 9.18 mmol.) dropwise over 5 minutes. The solution was further stirred at -78 0 C solution for 1 hour and treated with l-chloromethyl-naphthalene (1.62 g, 9.18 mmol.) in dry THF (10 mL). The solution was further stirred for 4 hours at -78 0 C and allowed to warm to room temperature overnight. Wet THF was added (50 mL) and the solution was concentrated under reduced pressure. The residue was dissolved in chloroform and washed with water. The organic phase was dried over sodium carbonate and concentrated. The crude product was purified by flash chromatography on silica gel (1:3:6 triethylamine:ethyl acetate:hexanes as eluent). The product WO 95/24200 PCT/US95/03099 127 containing fractions were pooled and concentrated to afford product (3.48 g as a viscous oil (m/e 539) which was used without further purification.

To a stirred solution of methoxymethyl indole prepared above (3.48 g 6.45 mmol.) in THF (30 mL) was added 2N HC1 (30 mL). The mixture was stirred at ambient temperature for 24 hours, and partitioned between diethyl ether and water. The aqueous phase was back extracted with diethyl ether (2 X 50 mL) and the combined organic phases were washed with brine, dried over sodium carbonate, and concentrated under reduced pressure. The residue was dissolved in THF (20 mL) and treated with 2N sodium hydroxide solution (6 mL). After 2 hours, the reaction mixture was extracted with chloroform (2 X 100 mL). The organic phase was dried over sodium carbonate and concentrated to afford product (2.68 g) as a viscous oil 495 To a stirred, cooled (0 0 C) solution of previously prepared formamidine (2.68 g, 5.41 mmol.) in ethanol (100 mL) was added water (12 mL) followed by acetic acid (12 mL) and hydrazine hydrate (22 mL). The reaction vessel was placed in the freezer (-10 0 C) for 72 hours. The mixture was warmed to ambient temperature and concentrated under reduced pressure.

The crude product was dissolved in chloroform (300 mL) and washed wit", water (3 X 50 mL). The organic phase was dried over sodium carbonate and concentrated to a viscous oil. The oil was dissolved in diethyl ether and treated with anhydrous HC1. The hydrochloride salt (1.50 g) was isolated by filtration. Recrystallization from ethanol (2 X) afforded material of constant rotation. Chiral HPLC confirmed enantiomeric purity as 95% ee. m/e=326 specific rotation 589 nM -40.21 (pyridine, C= specific rotation G 365 nM +80.43 pyridine, C=1) m ~l"~ys WO 95/24200 PCT/US95/03099 128 Analysis Calculated Found C 76.12 75.96 H 6.39 6.56 N 7.72 7.44 Example Preparation of 6-methyl-l- [(4-dimethylam.no-naphthalenyl)methyl]-1,2,3,4-tetrahydro-9H-pyrido [3,4-b]indole dihydrochloride monohydrate To a stirred, cooled (-78 0 C) suspension of methoxymethyl-triphenylphosphonium chloride 10.32 g, 30.1 mmol. in dry THF 150 mL was added n-BuLi solution 18.8 mL. 1.6 M, 30.1 mmol. dropwise by syringe. The orange suspension was stirred at -78 0 C for 15 min. A solution of 4-dimethylamino-l-naphthaldehyde 5.00 g, 25.1 mmol. in THF 75 mL was added to the ylide dropwise over 10 min.

The reaction mixture was gradually warmed to ambient temperature and stirred 14 H. Saturated ammonium chloride solution 100 mL was added and the mixture extracted with diethyl ether 3 X 50 mL The combined organic phases were dried over sodium sulfate.and concentrated under reduced pressure. Chromatography on silica gel, eluting with ethyl acetate/hexanes afforded product 5.43 g as a mixture of olefin isomers which was used without further purification.

A mixture of 5-methyltryptamine hydrochloride 695 mg, 3.3 mmol. and l-methoxy-4'-dimethylamino-benzostyrene 1.00 g, 4.4 mmol. in acetonitrile 20 mL and 1N HC1 solution 150 mL was heated to reflux for 96 H, with addition of 1 mL of cone HC1 added at 4 H. The reaction mixture was cooled to ambient temperature, neutralized with saturated aqueous potassium carbonate solution and extracted with chloroform. The combined organic phases were concentrated under reduced pressure and the residue chromatographed on silica gel 2.5% MeOH/chloroform/0.2% ~i Ce3l RIA~IIII WO 95/24200 PCT/US95/03099 129 as eluent The fractions containing product were pooled and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and treated with anhydrous HC1. The product was isolated as the dihydrochloride salt monohydrate( 1.22 g by filtration. mp. 231.3 0

C.

analysis: calculated found C 65.21 65.30 H 6.79 6.60 N 9.13 9.03 Example 66 7-methyl-8-bromo-1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole 'NH2 NH SHCI

'HCI

'HCI

Br

H

Br A 3.0 g sample of 6-methyl-7-bromo-lH-indole-3ethanamine hydrochloride was dissolved in warm water. A solution of glyoxylic acid monohydrate (1.0 g) in water was added. The solution was adjusted to pH 4 us4^g either potassium hydroxide or hydrochloride acid. A solid was suspended in water and concentrated HC1 was added slowly.

The mixture was boiled. A solid was collected, washed with water, and vacuum dried. The solid was partitioned between 1 N NaOH and chloroform. The organic portion was dried and concentrated to a residue which was chromatographed on silica gel using methanol in chloroform. The desired fractions were pooled and concentrated to a solid which was dissolved in methanol, treated with gaseous HC1, and diluted with ether.

A solid was collected, washed with ether, and dried.

Yield: 48% Melting Point: 321 0

C

Elemental Analysis: C 47.83; H 4.89; N 9.30.

WO 95/24200 PCT/US95/03099 130 Example 67 8-methoxy-1,2,3,4-tetrahydro-9H-pyrido 3,4b]-indole The desired product was prepared using substantially the process of Example 66, except that the starting material was 7-methoxy-lH-indole-ethanamine.

Melting Point: 207-209°C Elemental Analysis: C 60.17; H 5.56; N 8.60.

Example 68 8-methoxy-2(N)-propyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole NH N Pr O 1CI N

H

OMe OMe ,A sample of 8-methoxy-1,2,3,4-tetrahydro-9Hpyrido[3,4b]-indole was prepared substantially as described in Example 66. A 0.36 g sample of the indole was contacted with 1 g K 2 CO3 and the mixture was purged with nitrogen. A mL sample of CH3CN was added to the resulting mixture. A 0.12 mL sample of 1-iodopropane was added. The mixture was maintained under nitrogen and stirred in the dark. The resulting mixtvre was extracted. The organic phase was dried, evaporated, and chromatographed. The desired fractions were evaporated, taken up i ,to methanol:ethyl acetate. The resulting mixture was added to a stirring ether solution through which gaseous HC1 was bubbled. The resulting solid was vacuum dried, recrystallized, and evaporated to yield the desired product.

Yield: 0.10g Melting Point: 282-284 0

C

Elemental Analysis: C 64.45; H 7.67; N 9.91.

I

WO 95/24200 PCT/US95/03099 131 Example 69 8-methoxy-2 -methyl-1,2,3,4-tetrahydro-9H-pyrido [3,4b] indole NH NMe B "HCI "H C I

.I

N

N

OMe OMe A sample of 8-methoxy-l,2,3,4-tetrahydro-9Hpyrido[3,4b]-indole was prepared substantially as described in Example 66. The indole (1 NaOAc (0.34 NaBH3CN (0.53 methanol (50 mL), and HOAc (1.0 g) were stirred. A 1.36 g (37% in 10 mL methanol) sample of CH 2 0 was added to the indole mixture.

The reaction was quenched using an acid, then made basic, and extracted. The organic was dried, evaporated, and chromatographed. The desired fractions were evaporated and taken up into methanol/ethyl acetate. The resulting mixture was added to ethereal HC1. The resulting solid was collected and vacuum dried.

Yield: 0.84 g (79%) Melting Point: 291-294°C Elemental Analysis: C 62.06; H 6.97; N 11.32.

sR -alR1II II1 WO 95/24200 WO 9524200PCTUS95IO3099 132 Examrle 8-methoxy-2 -cYcloprOPYlmethyl-1, 2,3,4 -tetrahydro-9Hpyrido 4b] -indole INH

N

XN

H

The desired product was prepared using appropriate reagents and the pincess substantially as described in Example 69.

Yield: 88% Melting Point: 285-287 0

C

Elemental Analysis: C H 7.47; N 9.47.

IXAM 1 e 7 1 The desired product can be prepared using appropriate reagents and the process substantially as described in Example 69.

Yield: 48% Melting Point: 321 0

C

Elemental Analysis: C 47.83; H 4.89; N 9.30.

WO 95/24200 PCTIUS95/03099 133 Example 72 7,8-dimethyl-1,2,3,4-tetrahydro-9H-pyrido[3, 4b]-indole A 2.30 g sample of 6,7-dimethyl-lH-indoleethanamine was dissolved in a mixture of water and isopropanol with heating. A 1.03 g sample of glyoxilic acid monohydrate in 10 mL of water was added to the flask. The solution was allowed to cool and made basic by the addition of potassium hydroxide. The reaction was .tirred for 48 hours. The resulti g solid was isolated by filtration and washed with water. The solid was dissolved in 50 mL of water and the solution was acidified by the slow addition of concentrated HC1. Heating was initiated and an additional mL of concentrated HC1 was added. The resulting solid was isolated by decanting and dissolved in 10 mL of water. This solution was made basic by the addition of potassium hydroxide and extracted using 1:3 isopropanol:CHCl 3 Separation and concentration of the organic layer gave a viscous oil which was purified via chromatography. The oil was dissolved in ethyl acetate and gaseous HC1 was bubbled into the solution to form the hydrochloride salt. The solid hydrochloride salt was isolated by filtration and dried in a vacuum oven.

Yield: b4% Melting Point: 330 °C Elemental Analysis: C 65.75; H 7.29; N 11.62.

Example 73 6-methyl-8-bromo-1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole The desired 6-methyl-8-bromo-1,2,3,4-tetrahydro- 9H-pyrido[3,4b]-indole was prepared using appropriate reagents and the process substantially as described in Example 72.

Yield: 57% Melting Point: 346 0

C

Elemental Analysis: C 48.04; H 4.68; N 9.30.

WO 95/24200 WO 95/4200 CT[US95/03099 134 Example 74 6, 8-difluoro-1,2,3,4-tetrahydro-9u-pyrido[3,4bJ -indole The desired 6, 8-difluoro-1, 2,3, 4-tetrahydro-9Hpyrido[3..4b1J-indole was prepared using appropriate reagents and the process substantially as described in Example 72.

Yield: Melting Point: 350 0

C

Elemental Analysis: C 53.90; H 4.49; N 11. 23.

8-bromo-1, 2,3, 4-tetrahydro-9H-pyrido 4bJ-indole The desired 8-bromo-l, 2, 3,4-tetrahydro-9Hipyrido[3,4b]-indole was prepared using appropriate reagents and the process substantially as described in Example 72.

Yield: 4% Melting Point: 337.8 0

C

Elemental Analysis: C 46.17; H 4.26; N 9.52.

The following were prepared by the process substantially as described in Example 72.

8-fluoro-l, 2, 3 ,4-tetrahydro--9H-pyrido 4b] -indole Yield: 48% Melting Point: 329.5 0

C

Elemental Analysis: C 58.58; H1 5.43; N 12.37.

6-chloro-l, 2,3, 4-tetrahydro-9H--pyrido 4b3 -indole; Yield: 63% Melting Point: 317.9 0

C

6-bromo-l,2,3, 4-tetrahydro-9H-pyrido 4b] -indole; Yield: 19% Melting Point: 310.9 0

C

6-fluoro--, 2,3, &-tetrahydro-91i-pyrido 4b] -indole; Yield: 38% Melting Point: 316,,6 0

C

I

WO 95/24200 PCT/US95/03099 135 Yield: 54% Melting Point: 330 °C Elemental Analysis: C 65.75; H 7.29; N 11.62.

Example 76 7-methyl-8-chloro-1,2,3,4-tetrahydro-9H-pyrido[3,4b] -indole NH2 NH /HC1 "|C

CIHC

CI

The desired product was prepared using the process substantially as described in Example 1 except that the starting material was 6-methyl-7-chloro-lH-indole-3ethanamine hydrochloride.

Yield: 70 The resulting material was boiled in ethanol. The resulting product was collected, washed with ethanol, and vacuum dried.

Yield: 58% Melting Point: 330-334 0

C

Elemental Analysis: C 55.88; H 5.47; N 10.93.

The following were prepared using the process substantially as described above in Example 76.

7-methyl-8-chloro-l,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole Melting Point: 350-352 0

C

Elemental Analysis: C 55.65; H 5.68; N 10.39.

8-chloro-l,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole Melting Point: 335-337 0

C

Elemental Analysis: C 53.93; H 4.88; N 11.09.

WO 95/24200 PCT/US95/03099 136 7-bromo-8-methyl-l,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole Melting Point: 323-325 0

C

Elemental Analysis: C 4 7 .85; H 4.84; N 9.08.

Example 77 7-methyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole NH HiMI -HC1

*HCI

N N H H Br A sample of 7-methyl-8-bromo-l,2,3,4-tetrahydro- 9H-pyrido[3,4b]-indole was reacted with hydrogen in the presence of Pd/C, ethanol, and triethylamine. The resulting material was filtered, concentrated and extracted. The organic phase was dried, concentrated, and vacuum dried. The resulting solid was taken up into methanol and added to ethereal HC1. A white solid was collected, washed with Et 2 0, and vacuum dried.

Yield: 56% Melting Point: 310-312 0

C

Elemental Analysis: C 64.79; H 6.89; N 12.47.

Example 78 8-methyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]-indole The desired product was prepared using the process substantially as described in Example 77.

Yield: 46% Melting Point: 318-320 0

C

Elemental Analysis: C 64.53; H 6.94; N 12.43.

WO 95/24200 PCT/US95/03099 137 Example 79 7-bromo-1H-indole-3-ethanamine SCI H NH2

HHCI

NHNH

2

N

*HCI H Br Br A 25.8 g sample of 2-bromophenylhydrazine hydrochloride was partitioned between 1 N NaOH and chloroform. The organic layer was separated and the aqueous portion was extracted with chloroform. The combined organic extracts were dried (Na 2

SO

4 and concentrated to yield the free hydrazine as an oil.

The oil was stirred in 100 mL of methanol while 4chlorobutyraldehyde (12.3 g) was added. The resulting solution was transferred to a sealable tube and purged with nitrogen. The tube was sealed and the reaction mixture was heated in an oil bath maintained at 95 0 C for 14 hours. The resulting mixture was allowed to cool and concentrated to a residue which was partitioned between lN NaOH and chloroform.

The combined organic extracts were dried and concentrated to an oil. The oil was chromatographed on silica gel using a gradient of 0-10% methanol in chloroform. F.:actions containing product were concentrated to an oil which was taken up in a small amount of methanol and added to ethereal HC1. A solid was collected, washed with diethyl ether and vacuum dried at 50 0

C.

Yield: 7.32 g Yield: 23% 260-262 0

C

Elemental Analysis: C 43.55: H 4.41: N 10.03.

Example i PC Y- WO 95/24200 PCTIUS95/03099 138 .'ro-1H-indole-3-ethanamine EtO 1 HC *HC1

NHNH

2 H C l F F The desired 7-fluoro-lH-indole-3-ethanamine was prepared substantially as described in Example 81 infra.

except that 2-fluorophenylhydrazine hydrochloride (25.5 g) was used. Additionally, reverse phase HLC was required for final purification.

Yield: 4 g Melting point: 187-1890C Elemental Analysis: C 55.12; H 5.48; N 12.60.

Example 81 7-methoxy-1H- indole- 3 -ethanamine C1 H NH HC1

NHNH

2

.HCI

OMe OMe A 15.8 g sample of 2-methoxyphenylhydrazine hydrochloride and a 26.3 g sample of 4phthalimidobutyraldehyde diethyl acetal were stirred in ethanol. The mixture was heated at reflux for 2 hours. The reaction mixture was allowed to cool and was concentrated to a residue.

The resulting residue was dissolved in 750 mL ethanol and 15.5 g hydrazin: hiydrate was added. The mixture was heated at reflux for hours. A 70 mL sample of 5N HC1 was added and the mixture was allowed to cool. The cooled aR WO 95/24200 PCTIUS95/03099 139 mixture was concentrated to a residue. The residue was partitioned between IN NaOH and chloroform. The organic portion was separated and the aqueous portion was extracted with chloroform. The combined organic extracts were dried (Na 2

SO

4 and concentrated to an oil. The oil was chromatographed on silica gel using a gradient of 0-10% methanol in chloroform. Fractions containing product were concentrated to an oil which was taken up into a small amount of methanol and added to ethereal HC1. A solid was collected, washed with diethyl ether, and vacuum dried at 0 C to afford a white solid.

Yield: 7.5 g (37%) melting point: 198-200 0

C

Elemental Analysis: C 57.51; H 6.75; N 12.10.

Example 82 7-chloro-1H-indole-3-ethanamine O N

"HCI

CI H H Cl A 10.0 g sample of 2-chlorophenylhydrazine hydrochloride and 17.9 g of 4-phthalimidobutyraldehyde diethyl acetal were stirred in 200 mL ethanol with 1 mL 5 N HC1. The mixture was concentrated to a residue which was slurried in a small amount of methylene chloride. A yellow solid was collected and vacuum dried at 400C. The solid was stirred in 500 mL ethanol. Hydrazine hydrate (14 g) was added and the mixture was heated at reflux for 14 hours. A mL sample of 5 N HC1 was added and the mixture was he,-ted at reflux for 1 hour. The mixture wa- allowed to cool and was concentrated to a residue. The residue was partitioned -Rarpal-l--~ WO 95/24200 PCT/US95/03099 140 between 1 N NaOH and chloroform. The organic layer was separated and the aqueous layer was extracted with chloroform. The combined extracts were dried (Na 2

SO

4 and concentrated to an oil. The oil was chromatographed on silica gel using a gradient of 0-10% methanol in chloroform containing 0.2% ammonium hydroxide. Fractions containing product were concentrated to an oil which was taken up in a small amount of methanol and added to ethereal HC1. A solid was collected, washed with diethyl ether, and vacuum dried at 500C.

Yield: 3.2 g Melting Point: 227-229 0

C

Elemental Analysis: C 51.76; H 5.29; N 11.97.

Example 83 5-methyl-7-chloro-IH-indole-3-ethanamine N0 H HCI C1 The desired product was prepared substantially as described in Example 82.

Yield: 4.3 g (34%) Melting Point: 279-281 0

C

Elemental Analysis: C 54.05; H 5.85; N 11.33.

Example 84 1-H-Benz(G)indole-3-ethanamine -Pllraet 9 WO 95/24200 WO 9524200PCT/US95/03099 141 1-H-Benz indole-3-ethanamine was prepared using substantially the process described in Example 82.

Yield: 3.5 g (17%) Melting Point: 305-307 0

C

Elemental Analysis: C 68,43; H 6.30; N 11.08.

6-methyl-7-chlJoro-1H-indole-3-ethananine6-bromo-7-methyl-1Hindole-3 -ethananiine 6-ety-7cloo-Hinoe--ehnaie0a 6bo--methyl-1or-H-indole-3-ethanamine was preparedus substantially h epoes described in Eape8 sn aprpried sustartingl ma eas.rbdi xmpe8 sn WO 95/24200 PCT/US95/03099 142 Yield: 1.6 g (56%) Melting Point: 251 °C Elemental Analysis: C 45.85; H 4.97; N 9.71.

Example 86 6-methyl- in- indole- 3 -ethanamine NH2 'HC1 Hc

N

Br

H

A sample of 6-methyl-7-bromo-lH-indole-3ethanamine was contacted with Pd/C H 2 in the presence of ethanol and triethylamine. The resulting material was evaporated and partitioned between base/CHCl3. The organic phase was dried, concentrated, and dried. The resulting material was taken up into methanol and added to ethereal HC1. The resulting material was washed and vacuum dried.

Melting Point: 232-236 0

C

Elemental Analysis: C 62.84; H 7.24; N 13.20.

Example 87 5-methyl-7 -bromo-H-indole-3-ethanamine A sample of 5-methyl-7-bromo-lH-indole-3-ethanamine was prepared using appropriate starting materials and substantially the process described in Example 79.

Yield: 16% A 0.6 g sample of 5-methyl-7-bromo-lH-indole-3ethanamine hydrochloride salt was converted to the free base and chromatographed on silica. The desired fractions were pooled and evaporated. The resulting material was taken up into ethyl acetate, filtered, diluted with ether, and maleic acid in methanol. The product was crystallized using ether, filtered, and dried.

slk Illplr I sll~ep~Ap WO 95/24200 PCT/US95/03099 143 Yield: 67% Melting Point: 185-187 0

C

Elemental Analysis: C 49.09; H 4.85; N 7.71.

Example 88 6,7-dimethyl-1H-indole-3-ethanamine A sample of 6,7-dimethyl-1H-indole-3-ethanamine was piepared using appropriate starting materials and substantially the process described in Example 79. The 6,7dimethyl-lH-indole-3-ethanamine was purified by treating with

K

2

CO

3 and extracting with 3:1 CHCl3/isopropanol. The organic phase was dried, evaporated, and chromatographed. The desired fractions were pooled, evaporated, and mixed with ethyl acetate. The resulting material was diluted with ether and maleic acid in methanol. The solid was triturated in ether and dried.

Melting Point: 171-173 C Elemental Analysis: C 63.20; H 6.75; N 8.98.

Example 89 6-methyl-7-bromo-1H-indole-3-ethanamine A sample of 6-methyl-7-bromo-lH-indole-3-ethanamine was prepared using appropriate starting materials and substantially the process described in Example 79.

Yield: 8.6% The 6-methyl-7-bromo-1H-indole-3-ethanamine was dissolved in boiling ethanol and slowly cooled to room temperature. The solvent was reduced, the resulting material was filtered, and washed with ether. The resulting material was again filtered and washed with ether to afford the desired compound.

Melting Point: 288-290 0

C

Elemental Analysis: C 45.54; H 4.80; N 9.47.

For Examples 90 through 110, where applicable, diethylether was distilled from sodium benzophenone ketyl prior to use. All reactions were performed under a po tive

,~ONO

WO 95/24200 PCT/US95/03099 144 pressure of argon. 1 H-NMR and 13C-NMR data were recorded on e Bruker AC-200P (200 MHz). IR spectra were obtained on Nicolet 510 P-FT (film and KBr). Melting points were determined on a Bfchi apparatus and are not corrected. Analytical TLC was performed on Merck TLC glass plates precoated with F254 silica gel 60 (UV, 254 nm and Iodine). Chromatographic separations were performed by using 230-400 mesh silica gel (Merck). N-BOC-aziridines (2a-d) were prepared from the corresponding alkenes following standard procedures.

Preparation 2 Indole starting materials The indole starting materials (la, lb, and ic) infra. were purchased prepared according to Bartoli's procedure (Ib) [Bartoli, G. et al. Tetrahedron Lett., 1989, 2129] or (Ic) synthesized from 2-iodo-4,6-dimethylaniline The process is illustrated by the following Scheme: Scheme IV SiMea NH2 i M N Me NH S HiMeS Cul/DMF Me Cul/(PPh 3 2 PdCI 2 I2 e EtN Me Me Me 1C The 2-Iodo-4,6-dimethylaniline synthesis can be completed as follows: To a suspension of (24 mmol.), Cul (0.05 equiv.) and (PPh3)2PdC12 (0.05 equiv.) in 30 ml of dry triethylamine under Ar atmosphere was added trimethylsilylacetylene (1.1 equiv.) and the resulting mixture was stirred for 3 hours. Then, the solvent was eliminated under vacuum and the residue purified by flash chromatography using hexane/ethyl acetate as eluent to yield 6" in quantitative yield. A slurry of (23 mmol.) and Cul (2 equiv.) in 50 ml of dry dimethylformamide was heated for 2.5 h. under Ar atmosphere at 1000 C. After cooling down to room temperature the reactioi 'ure was L_ I L IP Fqll~LIC-- WO 95/24200 WO 9524200PCT11JS95/03099 145 filtered off and the solid washed twice with ether (20 ml.) The organic phase was washed with water (3x 5C dried over Na2SO4 and the solvent evaporated to dryness. The crude product was purified by flash chromatography using hexane/ethyl acetate as eluent to afford 1c (1.5 g., The process for preparing compounds of Examples through 107 is illustrated by the following Scheme:

.IX

b X".

C X.= 1. MeMgBr

HHC

2 Me 2 5.CuBr H.C H 12ann3O-- H VN 2bn=43 1"Boo 2c n~q=5V 4.HCI 5-Me a X'--5-Me n3O=3 (90) a X"=7-CI -7-CI bX-7C 5,7-diMe b X"-7-CI fl 3 0 -3 (91) c X'=-5-Me c X"=-5,7-M8 2 n 3

O

0 -3 d X"=7-CI d X"-5S-Me n3O-4 (92) e X"=-5,7-Me 2 a X"=7-CI n 3 O.:4 (93) f X=-5-Me fX"=5,7-Ma 2 n 3 O)-4 (94) g X"=7-CI g X=-5-Mo n3= h X"--5,7-M9 2 I X"=5S-Me I X'.-5-Me k Me I X"=5,7-Me 2 M X'=-5-Me n34R, R'-H n 30 4 R. R'=H (96) n 30 -3 R-3,4-(OMe) 2 Bn, R'=H (97) n 30 R=3,4-(OMe) 2 Bn, R'=H (98) n3O=3 R=3,4-(OMe) 2 Bn, R'=H (99) n3O=4 R=-3,4.(OM9) 2 Bn, R'=H (100) n 3 O-=4 Ft-3,4-(OMe) 2 8n, R=H (101) n 30 =4 R-3,4-(OMe) 2 Bn, R'=H(102) n3 0 O=5 R-3,4-(OtAe) 2 Bn, R'=H (103) n 30 =3 R=1-naphthylmethyI, R'=H (104) n 30 R=1-naphthylmethyl, R'=H (105) n3O--4 R=1-naphthylmethy, R'=H (106) n30=4 R, (107) N H. HCI or glyoxylic acid 1N HCI4f 1\ ome ome Example Trans-3-(2-amino-cyclop entyl)-5-methylin dole, hydrochloride To a suspension of the corresponding indole la mmol.) in 10 ml of anhydrous ether-under Ar atmosphere was added a 3M solution of methylmagnesium bromide (1.5 equiv.).

The resulting mixture was stirred for 45 min. at room temperature. Then, this mixture was cannulated to a slurry of WO 95/24200 PCT/US95/03099 146 Copper bromide-dimethylsulfide complex (0.2 equiv.) in ml. of dry ether under Ar atmosphere at -300 C. The reaction mixture was stirred for 30 min. at the same temperature.

After this time the mixture was cooled down to -780 C and the corresponding aziridine 2a (1.5 equiv.) dissolved in 10 ml.

of dry ether was added. The whole mixture was allowed to reach room temperature and stirring was kept overnight. The reaction was quenched with 10 ml. of a saturated solution of ammonium chloride. The layers were separated and the aqueous phase was extracted with ether/ethyl acetate (2x10 The combined organic extracs were dried over anhydrous sodium sulfate, the solvent was eliminated under vacuum and the resir'-e was purified by flash chromatography using hexane/ethyl acetate The corresponding N-BOC protected tryptamine was dissolved in dichloromethane/ether. The solution was saturated with dry hydrogen chloride and stirred overnight at room temperature. Finally, the solvent was evaporated and the crude title tryptamines purified by washing with dichoromethane/ether/methanol mixture The product was identified as the title compound (3a).

Yield: 85%. Mp: >200° C. H NMR (CD30D),8: 7.35 1H), 7.23-7.12 (m, 2H), 6.91 J= 7.5 Hz, 1H), 3.73 1H), 3.27 1H), 2.38-2.10 5H), 2.05-1.70 (mn, 4H). 13 C NMR (CD3OD), 8: 136.98, 128.93, 127.84, 124.27, 123.13, 119.01, 114.19, 112.37, 58.56, 43.93, 33.10, 31.30, 23.07, 21.73. IR (KBr): 3304, 2963, 1593, 1510, 1481, 800 cm- 1 MS 214 (M+-HC1, 28), 197 170 144 126 105 84 (100).

Eamp1le 91 Trans-3-(2-amino-cydopentyl)-7-chloroindole, hydrochloride The title compound (3b)was prepared using substantially the same procedure as described by Example however, the indole starting material was a compound of Formula lb.

Yield: 37%. Mp: >2000 C. 1 HNMR (CD30D),8:7,56 (d,J=7.7 Hz, 1H), 7.31 1H), 7.12 (d I 7.3 Hz, 1H), 7.0. J 7.8 Hz, 1H), 3.77 J= 7.9 Hz, 1H), 3.40-3.25 1H), 2.40-2.15 2H), 2.05-1.70 4H). 13C NMR (CD3OD), WO 95/24200 PCT/US9503099 147 135.48, 129.53, 124.28, 122.13, 120.79, 118.40, 118.02, 116.18, 58.55, 43.79, 33.32, 31.36, 23.11. IR (KBr): 3422, 3298, 3040, 2972, 2909, 1495 crn- 1 MS 235 (M+-C1, 100), 218 165 Examg~lpt 92 Trpns-3-(2-amino-cyclohexyl)-5-methylindole, hydrochloride The title compound (3d)was prepared using substantially the same procedure as cdescribed by Example Yield: 80%. Mp: >2000 C. 1HNMR(CD30D),5:7,44(s, 1H),7.27(d,J=8.3 Hz, 1H), 7.18 1H), 6.95 (dd, J 8.3 and 1.2 Hz, 1H), 3.55-3.40 1H), 2.86 (dt, J= 4.3 and 11.3 HIz, 1H), 2.42 3H1), 2.25-2.12 1H), 2.10-1.79 (mn, 1.75-1.40 3H). 13C NMR (CD30OD), 5: 136.97, 129.12, 127.74, 124.42, 123.73, 119.09, 114.77, 112.48, 56.22, 41.61, 34.75, 32.42, 26.93, 25.79, 21.73. IR (KBr): 3400, 3283, 3021, 2936, 2861, 1491 cm- 1 MS 229 100).

Example 94 Trans-3-(2-amino-cyclohexyl)-7-chloroindole, hydrochloride (3e) The title compound (3e)was prepared using substantially the same procedure as described by Example Yield: Mp: >2000 C. 1 HNMR(CD30D),:7,63(d,J=7.8Hz,1H), 7.35 1H), 7.14 J 7.4 Hz, 1H), 7.02 J 7.7 Hz, 1H), 3.60-3.40 1H), 3.08- 2.91 1H), 2.30-2.10 1H), 2.05-1.80 4H), 1.75-1.45 3H). 1 3 C NMR 8: 135.43, 129.41 125.00, 122.15, 120.87, 118.53, 118.09, 116.70, 56.12, 41.43, 34.74, 32.37, 26.80, 25.68. IR (KBr): 2938, 2859, 1429, 1341, 779, 735 cm- 1

MS

249 (Me-C1, 100).

Example 94 Trans-3-(2-amino-cyclohexyl)-5,7-dimethylindole, hydrochloride Trans-3-(2-amino-cyclopentyl)-5,7-dimethylindole, hydrochloride The title compound (3f) was prepared using substantially the procedure of Example 90; however, the indole was Ic and the aziridine was 2b.

Yield: 45%. Mp: >2000 C. 1HNMR(CD30D),5:7,27(s1H),7.19(s,1H), 6.77 1H), 3.42 (dt, J 11.0 and 4.2 Hz, 1H), 2.85 (dt, J= 1I.4 and 4.2 Hz, 1H), 2.44 3H), 2.39 3H), 2.30-2.10 1H), 2.08-1.83 (mi, 4H), 1.70-1.40 3H).

1 3 C NMR (CD3OD), 5: .36.39, 129.37, 127.39, 125.01, 123.56, 121,94, 116.78, WO 95/24200 PCT/US95/03099 148 115.16, 56.28, 41.70, 34.71, 32.40, 26.93, 25.80, 21.72, 16.93. IR (KBr): 3420, 3279, 3013, 2934, 2861, 1505 cm- 1 MS 242 (M+-HC1, 62), 225 199 184 171 158 (100), 145 128 115 97 (12).

Substantially the same procedure was used to prepare Trans-3- (2-amine-cyclopentyl) -5,7-dimethylindole, hydrochloride however, the aziridine was 2a.

Yield: 63 1 H NMR (DMSO-4), 6:10.8 1H), 8.12 (broad s, 3H), 7.30-7.20 2H), 6.70 1H), 3.70-3.55 1H), 3.55-3.20 1H), 2.38 3H), 2.36 3H), 2.30-2.10 2H), 2.00-1.60 4H).

Example Trans-10-methyl-2,3,4,4a,5,6,7,11c-octahydro-lH-indolo[2,3-clquinoline, hydrochloride A suspension of tryptamine hydrochloride (3a) (1.3 mmol.) in 10 ml. of distilled water was dissolved by heating.

To this solution glyoxylic acid (1.43 mmol.) in 1 ml. of water was added. Subsequently, a solution of KOH (1.3 mmol.) in 1 ml. of distilled water was slowly added to reach pH= 4.

The resulting solution was stirred at room temperature for 1 h. After this time, commercially available hydrochloric acid ml.) was added dropwise and the resulting mixture was refluxed for 30 min. Another portion of hydrochloric acid ml.) was added and the reaction further refluxed for min. Finally, the reaction mi::ture was cooled down to room temperature and filtered off. The title tetrahydro-bcarboline (4a) was subsequently washed with water and ethanol.

Yield: 81%. Mp: >2000 C. 1 HNMR(DMSO-), >11.0(s, 1H),9.92 (broad s, 1H), 9.68 (broad s, 1H), 7.38 1H), 7.23 J 8.3 Hz, 1H), 6.88 J= 7.8 Hz, 1H), 4.50-4.22 2H), 3.18-2.95 2H), 2.80-2.65 1H), 2.34 3H), 2.30-2.15 1H), 1.98-1.80 2H), 1.80-1.20 1 3 C NMR (DMSO-d6), 6: 134.75 127.31, 126.49, 125.64, 122.65, 119.11, 111.14, 108.82, 58.99, 37.18, 29.42, 28.84, 24.94, 24.43, 21.28. IR (KBr): 3391, 3266, 2936, 2861, 2801, 2762 cm- 1

MS

241 100).

WO 95/24200 PCT/US95/03099 149 Example6 Trans-8-chloro-2,3,4,4a,5,6,7,llc-octahydro-lH-indolo[2,3-c] quinoline, hydrochloride (4b) A suspension of tryptamine hydrochloride (3b) (1.3 mmol.) in 10 ml. of distilled water was dissolved by heating.

To this solution glioxylic acid (1.43 mmol.) in 1 ml. of water was added. Subsequently, a solution of KOH (1.3 mmol.) in 1 ml. of distilled water was slowly added to reach pH= 4.

The resulting solution was stirred at room temperature for 1 h. After this time, commercially available hydrochloric acid ml.) was added dropwise and the resulting mixture was refluxed for 30 min. Another portion of hydrochloric acid ml.) was added and the reaction further refluxed for min. Finally, the reaction mixture was cooled down to room temperature and filtered off. The title tetrahydro-bcarboline (4b) was subsequently washed with water and ethanol.

Yield: 45%. Mp: >2000 C. IHNMR(DMSO-c),5:>11.0(s,lH),10.05 (broad s, 1H), 9.87 (broad s, 1H), 7.58 J= 7.8 Hz, 1H), 7.16 J 7.6 Hz, 1H), 6.98 J= 7.9 Hz, 1H), 4.60-4.20 2H), 3.18-2.95 2H), 2.90-2.70 1H), 2.25- 2.18 IH), 1.98-1.75 2H), 1.65-1.20 13 C NMR (DMSO-d6),: :133,17 128.18, 127.23, 120.65, 120.03, 118.55, 115.78, 110.73, 58.74, 36.93, 29.16, 28.77, 24.88, 24.36. IR (KBr): 3422, 3231, 2936, 2861, 2760, 1429 cm- 1 MS 261 (M+-C1, 241 (100).

Example 97 Trans- dim ethoxybenzyl)-9-m ethyl-1,2,b, ,4a,5,6,10coctahydrocyclopenta[alpyrido [3,4-blindole, hydrochloride (4c) A suspension of the corresponding tryptamine hydrochloride (3a) (1 mmol) and the correponding 4-alkylidene-2-methyloxazolin-5-one (1.2 mmol) in 1N hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h.

After this time the reaction mixture was allowed to reach room temperature and 1L WO 95/24200 WO 9524200PCT/US95/03099 150 filtered off. The crude solid was purified by flash chromatography using dichloromethane/m~ Lhanol as eluent.

Yield: 88%. Mp: 187-191' C. 1 HNMR (DMSO-ct), 8: >11.0 10.38 (broad E, 1H), 9.25 (broad s, 1H), 7.50-7.15 (in, 3H), 7.15-6.80 (in, 5.0-4.70 (broad s, 1H), 3.75 6H), 3.40-2.80 2.49 3H), 2.20-1.70 (in, 4H), 1.55-1.30 (broad s, 1H). 13 C NM (DMSO-d6), 5: 148.73 147.90, 134.45, 130.24, 128.17, 127.64, 125.44, 123.03, 121.78, 118.43, 113.69, 111.95, 111.27, 110.64, 62.01, 57.50, 55.51, 37.49, 25.52, 25.14, 21.30, 20.73. IR (KBr): 3438, 3237, 2942, 1518, 1264, 1248 cm 4 1. MS 377 (Me-Cl, 100).

xamle 98 Trans-7-cbloro-5-(3,4-dimethoxybenzyl)-1,2,3,4,4a,5,6,lococtahydrocyclopenta[alpyrido [3,4-blindole, hydrochloride (4d) A suspension of the corresponding tryptamipne hydrochloride (3b) (1 mrnol) and the correponding 4- (1.2 mmol) in IN hydrochloric acid (3 ml.) was ref luxed under Ar atmosphere during 7? h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 52%. Mp: >2300 C dec. 1 HNMR(DMSO-c),8:>11.0(s,1), 10.40 (broad s, 1H), 9.30 (broad s, 1H), 7.60-7.42 (mn, 1H), 7.38-6.90 (in, 5K), 4.90- 4.75 (broad s, 1H), 3.78 3H), 3.76 3H), 3.40-3.00 2.15-1.80 (mn, 4H), 1.60- 1.35 (broad s, 1 3 C NMR (DMSO-d6), 8: 148.70, 147.91, 132.95, 131.78, 128.25, 127.02, 121.75, 121.11, 120.41, 117.96, 116.05, 113.54, 112.72, 111.99, 61.74,57.45, 55.50,37.27, 25.24, 25.07,2.0,77. IR (KBr): 3588, 3438, 1518, 1290 cm1 MS 398 (M++2-HCl, 40), 396 (M+-HC1, 100).

Example-99 Trans-5-(3,4-dimethoxybenzyl)-7,9-dimethyl-1,2,3,4,4a,5,6,10coctahydrocyclopenta[alpyrido 13,4-blindole, hydrochloride (4e) A suspension of the corresponding tryptamine hydrochloride (3c) (1 inmol) and the correponding 4- WO 95/24200 PCT/US9503099 151 (1.2 mmol) in 1N hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 87%. Mp: >2000 C. 1HNMR(DMSO-c),8:>11.0(s,1H),10.20 (broad s, 1H), 9.20 (broad s, 1H), 7.29 1H), 7.20-6.95 3H), 6.75 1H), 4.90- 4.70 (broad s, 1H), 3.78 6H), 3.30-2.90 2.48 3H), 2.34 3H), 2.10-1.70 4H), 1.60-1.30 (broad s, 1H). 13 C NMR (DMSO-d6), 8: 148.73 147.90, 134.01, 129.98, 128.31, 127.84, 125.10, 123.82, 121.75,120.42, 116.03, 113.58, 111.99, 111.21, 61.94, 57.62, 55.52, 37.60, 25.57, 25.17, 21.23, 20.75, 17.07. IR (KBr): 3447, 2910, 1520 cml. MS 391 (M+-C1,100), 239 Example 100 Trans-6-(3,4-dimethoxybenzyl)-10-methyl-2,3,4,4a,5,6,7,1 c-octahydro-1Hindolo[2,3-clquinoline, hydrochloride (4f) A suspension of the corresponding tryptamine hydrochloride (36) (1 mmol) and the correponding 4- (1.2 mmol) in 1N hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 85%. Mp: 197-2000 C. 1 HNMR(DMSO-),8:>11.0(s,1H),8.90 (broad s, 1H), 7.42 1H), 7.28 J= 8.3 Hz, 1H), 7.16 1H), 7.05-6.90 3H), 4.95-4.80 (broad s, 3.73 6H), 3.66-3.59 1H), 3.25-2.80 (in, 4H), 2.35 (s, 3H), 2.20-2.10 1H), 1.95-1.20 (mn, 6H). 1 3 C NMR (DMSO-d6), 5: 148.67 147.91, 134.92, 134.76, 129.72, 127.85, 127.45, 125.43, 122.91,121.85, 119.43, 113.59, 111.90, 111.30, 109.45,59.98, 55.47, 55.40, 37.08, 36.65, 29.48, 28.24, 24.94, 24.41, 21.32. IR (Kbi): 3439, 2936, 1516, 1464, 1453, 1265 cm- 1 MS 391 100).

-1 9, WO 95/24200 PCTIUS95/03099 152 Example 101 Trans-8-chloro-6-(3,4-dimethoxybenzyl)-2,3,4,4a,5,6,7,llc-octahydro-1Hindolo[2,3-clquinoline, hydrochloride (4g) A suspension of the corresponding tryptamine hydrochloride (3e) (1 mmol) and the correponding 4- (1.2 mmol) in 1N hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h, After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 47%. Mp: >2500 C. 1HNMR(DMSO-),5:>11.0(s,1H),9,75 (broad s, 1H), 8.90 (broad s, 1H), 7.64 J= 7.9 Hz, 1H), 7.20 J= 7.8 Hz 1H), 7.15-7.00 4H), 4.90-4.80 (broad s, 1H), 3.74 6H),3.70-3.60 1H), 3.25-2.85 4H), 2.20-2.15 1H), 1.95-1.25 6H). 13 C NMR (DMSO-d6), 8: 148.72, 148.00, 133.46, 131.35, 128.00, 127.08, 121.86, 121.13, 120.28, 119.01, 115.99, 113.41, 111.98, 111.66, 59.62, 55.53, 55.42, 54.98, 37.24, 36.49, 29.23, 28.25, 24.88, 24.34. IR (KBr): 3428, 2938, 1518, 1250 cm-1. MS 410 (M+-HC1, 100).

Example 102 Trans-6-(3,4-dimethoxybenzyl)-8,10-dimethyl-2,3,4,4a,5,6,7,llc-octahydro-1Hindolol2,3-cquinoline, hydrochloride (4h) A suspension of the corresponding tryptamine hydrochloride (3f) (1 mmol) and the correponding 4- (1.2 mmcl) in LN hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 78%. Mp: 198-2020 C. 1HNMR(DMSO-cb),8:10.88(s,1H),9.81 (broad s, 1H), 8.78 (broad s, 1H), 7.24 1H), 7.20 1H), 7.10-6.90 2H), 6.73 1H), 4.90-4.75 (broad s, 1H), 3.74 6H), 3.25-3.10 2H), 3.10-2.80 2H), 2.45 3H), 2.32 3H), 2.20-2.10 1H), 2.00-1.80 3H), 1.60-1.10 3H).

IC IIC p ap 'Ir~eP-qlg WO 95/24200 WO 95/4200 CT[US95103099 153 1 3 C NM (DMSO-d6), 5:148.65 147.87, 134.44, 129.55, 128.17, 127.59, 125.13, 123.68, 121.90, 120.36, 117.05, 113.64, 111.89, 110.04, 59.89, 55.78, 55.41, 37.17, 36.56, 29.47, 28.21, 24.94, 24.43, 21.26, 17.09. IR (KBr): 3450, 2936, 1516, 1493, 1264, 1240 cm- 1 MS 405 (W~-0,100).

Example-103 Trans-7-(3,4-dimethoxybenzyl)-11-methyl-1,2,3,4,5,5a,6,7,8,12adecahydrocycloheptalapyrido[3,4-blindole, hydrochloride (4i) A suspension of the corresponding tryptamine hydrochloride (3g) (1 inmol) and the correponding 4- (1.2 inmol) in iN hydrochloric acid (3 ml.) was ref luxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 35%. Mp: 187-1900 C. 1HNM(DMSO-),:>11.0(s1H),9.66 (broad s, 1H), 7.29-7.25 2H), 6.92 J= 7.8 Hz, 1H), 6.81 J= 8.2 Hz, 1H)/ 6.65-6.56 4.80-4.70 (broad s, 1H), 3.66 3H), 3.43 3H), 3.00-2.90 (in, 1H),2.90-2.70 (in, 1H), 2.35 3H), 2.35-2.20 (mn, 1H), 1.80-1.30 (mn, 8H), 0.85-0.65 1H). 13 C NMR (DMSO-d6), 8: 148.56 147.96, 135.12, 128.81, 128.05, 127.27, 125.32, 123.09, 121.73, 118.97, 113.32, 111.85, 111.31, 110.51, 55.60, 55.08, 54.97, 51.48, 36.97, 36.24, 32.74, 31.88, 26.37, 24.88, 24.14, 21.30. FR (KBr): 3414, 3343, 2932, 2859, 1516, 1265 cnv- 1 MS 405 100), 335 ExampleC 104 Trans- 9-methyl-5- (1-naphthylme thyl)-1,2,3,4,4a,5,6,10coctahydrocyclopentalialpyrido 13,4-blindole, hydrochloride (4j) A suspension of the corresponding tryptamine hydrochloride (3a) (1 znmol) and the correponding 4- (1.2 mmol) in iN hydrochloric acid (3 ml.) was ref luxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid WO 95/24200 WO 9524200PCI, US95/03099 154 was purified by flash chromatog' %phy using dichioromethane/inethanol (9 as eluent.

Yield: 78%. Mp: >2000 C. 1E-1NM(DMSO-ct),:>11.0(s,1H),10.45 (broad s, 1H), 9.03 (broad s, 1H), 8.46 J= 7.9 Hz, 1H), 8.12-7.90 (in, 3H), 7.70- 7.40 (mn, 3H), 7.40-7.25 (mn, 2H), 6.96 J= 8.0 Hz, 1H), 5.15-4.90 (broad s, 1H), 4.45-4.30 (mn, 1H), 3.65-3.50 3.15-2.95 (in, 1H), 2.38 2.00 -4.70 (in, 4H), 1.60-1.35 (broad s, 1H). 1 3 C NMR (DMSO-d6), 6: 134.59,133.86, 131.63, 131.32, 129.92, 129.18, 128.86, 128.07, 127.74, 126.38, 125.96, 125.83, 125.48, 124.08, 123.20, 118.52, 111.31, 110.97, 61.78, 5r5.76, 37.40, 35.13, 25.49, 25.12, 21.32, 20.67. IR 3445, 3231, 2949, 2878, 2780, 793 cm1 MS 367 100).

Example 105 Trans-1-methyl-6-(l-naphthylmethyl)-2,3,4,4a,5,6,7,llc-octahydro-1Hindololi2,3-clquinoline, hydrochloride (4k) A suspension of the corresponding tryptainine hydrochloride (3d) (1 inmol) and the correponding 4- (1.2 inmol) in IN hydrochloric acid (3 ml.) was ref luxed under Ar atmosphere during 72,h. After this time tthe reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography vsing dichloromethane/rnethanol as eluent.

Yield: 80%. lMp: >2 000 C. 1 H NMR (DMSO-d6), 5: >11.0 1H), 8.40 (d, J= 7.8 Hz, 1H), 8.01 J= 7.5 FHz, 1H), 7.92 J= 8,2 Hz, 1H), 7.74 J= 6.8 Hz, 1H), 7.70-7.40 (nm, 4H), 7.35 J= 8.4 Hz, 1W), 6.97 J= 8.2 Hz, 1H), 5.15-4.90 (broad s, 1H), 4.50-4.30 (in, 1H), 3.50-3.10 2H), 3.10-2.82 (in, 2H), 2.38 3H-), 2.10-1.20 (in, 7H). 13 C NMR (DMSO-d6), 6: 135.05,134.90, 133.85, 131.79, 13-1.28, 129.36, 128.93, 128.07, 127.56, 126.33, 125.94, 125.83, 125.41, 124.02, 123.10, 119.54, 111.27, 109.61, 59.72, 53.97, 36.73, 35.27, 29.47, 28.37, 24.92, 24.36, 21.34. IR (KBr): 3447, 3235, 2936,2857, 1450, 790 cm-1. MS 381 100).

WO 95/14200 PCTIUS95/03099 155 Example 106 Ti'rans-8,10-dimethyl-6-(1-naphthylmethyl)-2,3,4,4a,5,6,7,11llc-octahydro-1Hindolo[2,3-c]quinoline, hydrochloride (41) A suspension of the corresponding tryptamine hydrochloride (3f) (1 mmol) and the correponding 4- (1.2 mmol) in 1N hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room tempe :,ture and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methano1 as eluent.

Yield: 77%. Mp: >2000 C. 1 HNMR (DMSO-c), 6: >11.0 1H),10.11 (broad s, 1H), 8.52 J= 8.2 Hz, 1H), 8.35 (broad s, 1H), 8.02 J= 7.3 Hz, 1H), 7.92 J= 7.9 Hz, 1H), 7.82 J= 6.9 Hz, 1H), 7.71-7.46 3H), 7.29 1H), 6.78 1H), 5.10-4.90 (broad s, 1H), 4.70-4.50 1H), 3.40-3.20 2H), 3.10-2.80 (m, 2H), 2.51 3H), 2.34 3H), 2.05-1.90 1H), 1.80-1.70 2H), 1.60-1.20 (nm, 4H). 1 3 C NMR (DMSO-d6), 6: 134.57,133.87, 131.95, 131.42, 129.29, 129.11, 128.81, 128.04, 127.71, 126.21, 125.91, 125.83, 125.14, 124.46, 123.91, 120.46, 117.14, 110.25, 59.65, 54.03, 36.66, 35.25, 29.47, 28.32, 24.94, 24.35, 21.26, 17.30. IR (KBr): 3449, 2934, 2859, 2791, 1449, 779 cm-1. MS 395 100).

Example 107 Trans-s iro-6,6-[2-(3,4-dimethoxy)-1,2,3,4-tetrahydronaphthyll-10-methyl- 2,3,4,4a,5,6,7,ll1a-octahydro-1H-indolo2,3-cquinidine, hydrochloride (4m) A suspension of the corresponding tryptamine hydrochloride (3a) (1 mmol) and the correponding 4- (1.2 mmol) in 1N hydrcchloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

I ~n.

WO 95/24200 PCTIUS95/03099 156 Epimeric mixture. Yield: 89%. 1HNMR(DMSO-d),6: >11.0(s,1H), 10.12 (broad s, 1H), 8.72 (broad s, 1H), 7.42 1H), 7.21 11), 6.90-6.60 3H), 3.75 3H), 3.71 3H), 3.30-2.80 5H), 2.35 3H), 2.00-1.20 6H). 1 3

C

NMR (DMSO-d6), 147.44, 134.84, 134.32, 133.98,127.42, 126.53, 126.35,125.25, 125.13, 123.60, 123.25, 122.98, 119.56, 119.43, 112.05, 111.48, 111.27, 108.78, 108.60, 57.E2, 57.50, 56.07, 55.56, 36.40, 31.91, 30.74, 29.39, 29.21, 28.73, 28.41, 24.92, 24.38, 23.83, 21.30. IR (KBr): 3440, 2950, 1518, 1200, 1110, cm- 1 MS 417 (M+-C1, 100).

Example 108 Trans-1-(3,4-dimnethoxybenzyl)-3,4,6-trimethyl-1,2,3,4-tetrahydro-9H-pyrido[3,4b]indole, hydrochloride (4n) Trans-3-(2-amiine-1,2-dimethylethyl)-5-methylindole, hydrochloride(3h) was prepared using substantially the procedure of Example 90; however, the aziridine was 2c, Yield: 71%. 1H NMR (CD30D),6: 7,45 1H), 7.32 J= 8.3 Hz, 1H), 7.19 (s, 1H), 7.00 (dd, J= 8.4 and 1.5 Hz, 1H), 3.66 J 6.9 Hz, 1H), 3.28 J= 7.3 Hz, 11), 2.47(s, 3H), 1.48 J= 7.2 Hz, 31H1), 1.38 J= 6.6 Hz, 31). 1 3 C NMR (CD30D), 8: 136.89, 129.19, 127.68, 124.46, 123.69, 119.09, 115.41, 112.44, '3.51, 36.62, 21.71, 17.06,16.49.

A suspension of the corresponding tryptamine hydrochloride (3h) (1 mmol) and 6,7-dimethoxytetralin-2-one (1.2 rmol) in 1N hydrochloric acid (3 ml.) was refluxed under Ar atmosphere during 72 h. After this time the reaction mixture was allowed to reach room temperature and filtered off. The crude solid was purified by flash chromatography using dichloromethane/methanol as eluent.

Yield: 32%. Mp: 195-1990 C. 1 HNMR(DMSOC-), >11.0(s,11),9.40 (broad s, 1H 8.90 (broad s, 1H), 7.40 1H), 7.30 J= 8.2 Hz, 1H), 7.08 1H), 6.96-6.90 3H), 4.90-4.80 (broad s, 1H), 3.73 3H),3.72 3H), 3.70-3.60 (m, 2H), 3.20-3.00 3H), 2.37 3H), 1.46 (broad s, 3H), 1.40 (broad s, 3H), 13C NMR (DMSO-d6), 5: 148.66 147.93, 135.00, 129.21, 127.40, 125.40, 122.97, 121.82, 119.07, 113.56, 111.95, 111.24, 110.34, 57.32, 55.43, 55.33, 54.60, 36.46, 32.56, 21.24, -L eLL~ WO 95/24200 WO 9524200PCTfUS95/03099 157 17.06, 15.92. IR (KBr): 3438, 2936, 1518, 1464, 1265, 1242, 1040 cinl. MS 365 Cl, 100).

Example 109 Cis-3-(2-amine-cyclohexyl)-5-methylindole, hydrochloride Cis..6-(3,4-dimethoxybenzyl)-1O-methyl-2,3,4,4ai,5,6,7,llc-octahydro-IHindolo[2,3-clquinoline, hydrochloride (4o) The title compound (3i) was prepared following tho procedure described by Scmuszkovicz, J. et al. Tetrahedron, 1991, 47, 8653 starting from 5-methylindole (1a).

Mp: 86-90- C. 1 HNYR (CD3OD), 5:7,38 1K), 7.26 (dJ=8.3 Hz, 7.11 6.96 J 8.2, 1H), 3.90-3.70 (in, 1K), 3.55-3.38 (in, 1H), 2.42 3H), 2.40- 2.35 (in, 1K), 2.10-1.79 (mn, 4H), 1.75-1.50 (mn, 3H). 1 3 C NMR (CD3OD), 5: 136.75, 129.27, 127.88, 124.63, 123.51, 118.71, 114.49, 112.34, 52.60, 36.79, 29.52, 26.44, 25.85, 21.68, 21. 00. IR (KBr): 3401, 3017, 2932, 286i3, 1561, 1489 cin1. MS 229 (M-ci, 100).

The process for preparing the final product (4o) is illustrated by the following Scheme: 0 Meo i. .4 H M NH 2 .HCI -,OC~e M NH.HCI 4o OMO 31 4 Mp: 167-171O C. IK NM (DMSO-d 8: >11.0 1H), 8.8 7 (broad s, 7.29- 7.20 (mn, 3H), 7.12-6.85 (in, 3K), 4.95-4.80 (broad s, 1H), 3.76 3H), 3.75 3), 3.70-3.60 3.25-3.00 (in, 1K), 2.36 3H), 2.40-2.00 1.95-1.20 (nW. 6H). 13

C

NUR (DMSO-d6), 5: 148.67 147.87, 134.80, 128.71,19$,1.43, 127.63, 125.45, 123.32, 121.75, 117.82, 123.59, 111.91, 111.30, 111.34, 56.99, 55.46, 55.12, 36.12,36.65, 28.42, 27.49, 24.94, 24.39, 21.23, 19.17. 1K 3439, 292-4, 1516, 1263 cin1. MS 390 (M+-CIH, 100).

As noted above, the compounds of the present invention are useful in blocking the effect of serotonin or other agonists at 5-HT2A, 5-HT2B and/or 5-HTIc receptors.

Thus, the present invention also provides a method for blocking 5-HT2A, 5-HT2B3 or 5-HTic receptors in manmmals comprising administering to a mammal requiring blocking of a WO 95/24200 PCT/US95/03099 158 5-HT2A, 5-HT2B, or 5-HTIc receptor, respectively, a receptor blocking dose of a compound of the invention.

One particularly useful embodiment of this invention is that it provides selective ligands for the 5-HT2B receptor. Compounds with a high affinity for the 5-HT2B receptor generally are cross-reactive with the 5-HT2c receptor as well. Now 5-HT2B receptors can be selectively modulated using compounds of this invention at rates set forth above for blocking the effects of agonists at 5-HT2B receptors. The selective affinity may provide treatments with fewer side effects and will facilitate the development of additional therapeutic agents.

Compounds exhibiting activity at the 5HT2B receptor are useful for treating disorders related to the modulation of the 5HT2B receptor. For example, compounds having 5HT2B antagonist activity reduce the spasticity of the colon.

Thus, these compounds are useful for the treatment of functional bowel disorders including irritable bowel syndrome and irritable bowel syndrome-related symptoms. The antispasmodic effect of such compounds can reduce abdominal pain associated with functional bowel disorders.

Additionally, the 5HT2B receptor is localized in other organs such as the brain, bladder, blood vessels, stomach, and uterus, indicating that additional conditions are 5HT2B mediated.

Compounds demonstrating activity at the 5HT2A receptor can be utilized in the treatment or prevention of conditions related to modulation of the 5HT2A receptor.

Examples of such conditions include hypertension, sleep disorders, hallucinogenic activity, psychosis, anxiety, depression, thermoregu2ation, feeding disorders, and hypotension. Leonard, International Clinical Psvchopharmacolov, 7, 13-21 (1992).

The term "receptor blocking dose", means an amount of compound necessary to block a targeted receptor, selected from the group consisting of 5-HT2A, 5-HT2B, and WO 95/24200 PCTIUS95/03099 159 receptor in a mammal. The active compounds are effective over a wide dosage range. For example, dosages per day will normally fall within the range of about 0.05 to about 250 mg/kg of body weight. In the treatment of adult humans, the range of about 0.5 to 100 mg/kg, in single or divided doses, is preferred. The ranges of about 5 mg/kg to about 60 mg/kg and about 10 mg/kg to about 50 mg/kg are especially preferred. However, it will be understood that the amount of the compound actually administered will be determined by a physician in light of the relevant circumstances, including the condition to be treated, the choice of compound to be administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the chosen route of administration, and therefore the above dosage ranges are not intended to limit the scope of the invention in any way. The compounds may be administered by a variety of routes such as oral, transdermal, s. bcutaneous, intranasal, intramuscular, and intravenous routes.

While it is possible to administer a compound of the invention directly without any formulation, the compounds are preferably employed in the form of a pharmaceutical formulation comprising a pharmaceutically acceptable excipient and at least one compound of the invention. Such compositions contain from about 0.1 percent by weight to about 90.0 percent by weight of a present compound. As such, the present invention also provides pharmaceutical formulations comprising a compound of the invention and a pharmaceutically acceptable excipient therefor.

In making the compositions of the present invention, the active ingredient is usually mixed with an excipient which can be a carrier, or a diluent or be diluted by a carrier, or enclosed within a carrier which can be in the form of a capsule, sachet, paper or other container.

When the carrier serves as a diluent, it can be a solid, semi-solid, or liquid material which acts as a vehicle, excipient, or medium for the active ingredient. Thus, the composition can be in the form of tablets, pills, powders, c- a I L Cc- ~t WO 95/24200 PCT/US95/03099 160 lozenges, sachets, cachets, elixirs, emulsions, solutions, syrups, suspensions, aerosols (as a solid or in a liquid medium), and soft and hard gelatin capsules.

The compounds of the invention may be delivered transdermally, if desired. Transdermal permeation enhancers and delivery systems, including patches and the like, are well known to the skilled artisan.

Examples of suitable carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, tragacanth, gelatin, syrup, methyl ce.lulose, methyl- and propylhydroxy- benzoates, talc, magnesium stearate, water, and mineral oil. The formulations may also include wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents. The formulations of the invention may be formulated so as to provide quick, sustained, or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art.

The compounds of this invention may be delivered transdermally using known transdermal delivery systems and excipients, Most preferrably, a compound of this invention is admixed with permeation enhancers including, but not limited to, propylene glycol, polyethylene glycol monolaurate, and azacycloalkan-2-ones, and incorporated into a patch or similar delivery system. Additional excipients including gelling agents, emulsifiers, and buffers may be added to the transdermal formulation as desired.

For oral administration, a compound of this invention ideally can be admixed with carriers and diluents and molded into tablets or enclosed in gelatin capsules.

The compositions are preferably formulated in a unit dosage form, each dosage containing from about 1 to about 500 mg, more usually about 5 to about 300 mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for e~s 19a~ mm -I WO 95/24200 PCT/US95/03099 161 human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical carrier.

The active compounds are effective over a wide dosage range. For example, dosages per day will normally fall within the range of about 0.05 to about 250 mg/kg of body weight. In the treatment of adult humans, the range of about 0.5 to 100 mg/kg, in single or divided doses, is preferred. The ranges of about 5 mg/kg to about 60 mg/kg and about 10 mg/kg to about 50 mg/kg are especially preferred.

However, it will be understood that the amount of the compound actually administered will be determined by a physician in light of the relevant circumstances, including the condition to be treated, the choice of compound to be administered, the age, weight, and response of the individual patient, the severity of the patient's symnptoms, and the chosen route of administration, and tharefore the above dosage ranges are not intended to limit the scope of the invention in any way. The compounds may be administered by a variety of routes such as oral, transdermal, subcutaneous, intranasal, intramuscular, rectally, and intravenous routes.

Pharmaceutical compositions of a compound of the present invention or its salts or solvates are most preferrably produced by formulating an active compound in unit dosage form with a pharmaceutical carrier. Some examples of unit dosage forms are tablets, pills, powders, aqueous and non-aqueous oral solutions and suspensions, transdermal delivery devices and patches, and parenteral solutions packaged in containers containing either one or more unit dosages and may be capable of being subdivided into individual doses. Some examples of suitable pharmaceutical ,arriers and/or diluents include gelatin capsules, sugars including lactose and sucrose, starches such as corn starch and potato starch, cellulose derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, WO 95/24200 PCT/US95/03099 162 and cellulose acetate phthalate, gelatin, talc, stearic acid, magnesium stearate, vegetable oils such as peanut oi±, cottonseed oil, sesame oil, olive oil, corn oil, and oil of theobroma, propylene glycol, glycerin, sorbitol, polyethylene glycol, water, agar, alginic acid, isotonic saline, phosphate buffer solutions, lactic acid, glycolic acid, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, magnesium stearate, croscarmellose, alginic acid, sodium starch glycolate, lauryl sulfate. as well as other compatible substances used in pharmaceutical formulations. The active compound may be prepared as a microparticle using biodegradable polymers or other known methods. The composition may be prepared using known formulation technology to provide a rapidly dissolving, sustained release, or targeted delivery compositions. The compositions of this invnetion may contain other components such as coloring agents, flavoring, and/or preservatives.

The compositions may contain other therapeutic agents, for example an antacid or analgesic.

The article of manufacture will include packaging material. Packaging material will preferably include a container. The preferred container and packaging material can be selected using the characteristics of the compound to be packaged. For example, the preferred container may be glass, plastic, foil, sealed bubble packaging, clear, amber, and may incorporate other known pharmaceutical packaging technology. The packaging may include features such as cotton, silica or other drying agents, and/or a measuring device. The article of manufacture shall include a label indicating that the composition is useful for the treatment of an a condition associated with 5-HT2B receptor stimulation malfunction. Most preferably, the condition is selected from the group consisting of urinary incontinence, bladder dysfunction, uterine dysfunction, cardiovascular disorder, and respiratory disorder.

In order to illustrate more fully the operation of this invention, the following formulation examples are WO 95/24200 PCT/US95/03099 163 provided. The examples are illustrative only, and are not intended to limit the scope of the invention. The formulations may employ as active compounds any of the compounds of the present invention.

I L WO 95/24200 PCT/US95/03099 164 Formulation 1 Hard gelatin capsules are prepared using the following ingredients: Concentration Amount Per by Weight Cansule (nercent) 6-ethyl-8-chloro-l- [(3,4-dimethoxyphenyl)methyl] -1,2,3,4-tetrahydro-9Hpyrido[3,4-b]indole hydrochloride 250 mg 55.0 starch dried 200 mg 43.0 magnesium stearate 10 ma 460 mg 100.0 The above ingredients ar mixed and filled into hard gelatin capsules in 460 mg quantities.

WO 95/24200 PCTIUS95/03099 165 Formulation 2 Capsules each containing 20 made as follows: Amount Per Capsule 6-methyl-8-ethyl-l- [(3-bromo-4-chloro-phenyl) -methyl)-1,2,3,4-tetrahydro- 9H-pyrido[3,4b]indole (Z)-2-butenedioate mg of medicament are Concentration by Weight (percent) 10.0 44.5 44.5 20 mg 89 mg starch microcrystalline cellulose magnesium stearate 89 mg 2 mQ 200 mg 100.0 The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 45 mesh U.S. sieve and filled into a hard gelatin capsule.

WO 95/24200 WO 9524200PCr1US95103099 166 Formulation 3 Capsules each containing 100 made as follows: Amount Per mg of medicament are Concentration by Weight (ioercent) luoro-6-methyl- 1- (1-(3-methylaminophenyl) methyl) -1,2,3,4tetrahydro-91i-pyrido- [3,4-blindole butenedioate polyoxyethyl ene sorbitan inonooleate starch powder 100 mg 30.00 50 mg 2 50 maT 400 mg 0 .02 69.9 100. 00 The above ingredients are placed in an empty gelatin capsule.

thoroughly mixed and WO 95/24200 PCT/US95/03099 167 Formulation 4 Tablets containing 10 mg of made as follows: Amount Per Capsule active ingredient are Concentration by Weight (percent) 6-fluoro-8-phenoxy- 1-(1-(4-ethoxy-phenyl)methyl)-1,2,3,4tetrahydro-9H-pyrido- [3,4-b]indole butenedioate starch microcrystalline cellulose polyvinylpyrrolidone (as 10% solution in water) sodium carl' ,xymethyl starch magnesium stearate talc 10 mg 45 mg 35 mg 4 mg 4.5 mg 0.5 mg 1 ma 100 mg 10.0 45.0 35.0 100.0 The active ingredient, starch and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly.

The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granule so produced is dried at 500-60 0 C and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh U.S. sieve, are then added to WO 95/24200 PCT/US95/03099 168 the granule which, after mixing, is compressed on a tablet machine to yield a tablet weighing 100 mg.

Formulation A tablet formulation may be ingredients below: Amount Per Capsule 5,6-difluoro- 1-1(-(3-dimethylamino-phenyl)methyl) -1,2,3,4tetrahydro-9H-pyrido- [3,4-b]indole butanedioate 250 mg prepared using the Concentration by Weight _(ercent) 38.0 60.0 microcrystalline cellulose silicon dioxide fumed stearic acid 400 mg 10 mg 5 ma 665 mg 100.0 The components are blended and compressed to form tablets each weighing 665 mg.

WO 95/24200 PCT/US95/03099 169 Formulation 6 Suspensions each containing 5 mg of medicament per ml dose are as follows: per 5 ml of suspension 3-methyl-5-chloro-6-methyl- 1- (3-dimethylamino-phenyl) methyl)-1,2,3,4tetrahydro-9H-pyrido- [3,4-b]indole butenedioate 5 mg sodium carboxymethyl cellulose 50 mg syrup 1.25 ml benzoic acid solution 0.10 ml flavor q.v.

color q.v.

water q.s. to 5 ml The medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethylcellulose and syrup to form a smooth past3. The benzoic acid solution, flavor and color is diluted with some of the water and added to the paste with stirring. Sufficient water is then added to produce the required volume.

WO 95/24200 PCT/US95/03099 170 Formulation 7 An aerosol solution is prepared containing the following components: Concentration by Weight (percent) 5-propyl-6-ethyl- 1-[(3,4-dimethoxy-phenyl)methyl)-1,2,3,4tetrahydro-9H-pyrido- [3,4-b]indole hydrochloride ethanol Propellant 22 (chlorodifluoromethane) 0.25 29.75 70.00 100.00 The active compound is mixed with ethanol and the mixture added to a portion of the Propellant 22, cooled to 0 C and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted further with the remaining amount of propellant. The valve units are then fitted to the container.

WO 95/24200 PCT/US95/03099 171 Formulation 8 Injectables may be prepared as follows: Amount Per Batch 6-(1-methylethyl)- 1,2,3,4-tetrahydro-l- (1-(4-dimethylaminonaphthalenyl) -methyl)-9H-pyrido[3,4b]indole (Z)-2-butenedioate mg Devazepide for Injection q.s The compound or a suitable salt thereof is dissolved in, for example, ethanol, and passed through a 0.2 micron filter. Aliquots of filtered solution are added to ampoules or vials, sealed and sterilized.

WO 95/24200 PCT/US95/03099 172 Formulation 9 Tablets containing 10 mg of made as follows: Amount Per Tablet active ingredient are Concentrati ii by Weight (nercent) 7,8,9,10-tetrahydro- 10-(1-(2dimethylaminonaphthyleneyl)methyl)-llH-benzo[g]pyrido[3,4-b]indole (Z)-2-butenedioate 6 g corn starch microcrystalline cellulose Sterotex Powder HM Purified Water 200 g 46 g 4 g 78.0 18.0 300 mL 100 mg 100.0 The active ingredient, starch and cellulose are combined together in a planetary mixer and mixed for 2 minutes. Water is added to the combination and mixed for 1 minute. The resulting mix is spread on trays and dried in a hot air oven at 50 0 C until a moisture level of 1 to 2 percent is obtained. The dried mix then milled with a Fitzmill through a #RH2B screen, and added back to the milling mixture. The mixture is drum rolled for 5 minutes.

Compressed tablets of 50 mg, 150 mg, and 200 mg are formed with appropriate sized punches.

WO 95/24200 WO 9524200PCT1US95i03099 173 Formulation Capsules are prepared using the following ingredients: Concentration Amount Per by Weight Cap-a-1-2-(percent) +-)6-methyl-1-C1-(3-ethylamino naphthalenyl) -1-ethyl) 1,2,3, 4-tetrahydro-9Hpyrido[3,4-blindole (Z) 2-butenedioate 200 mng 49.0 lactose USP 200 mg 49.0 Serotex Powd r 10 g2.

410 mg 100.0 The above ingredients are mixed and filled into hard gelatin capsules in 410 mg quantities.

WO 95/24200 WO 9524200PCTfUS95/ 03099 174 Formulation 11 Hard gelatin capsules are prepared using the following ingredients: Amount Per Concentration by Weight (percent) (1naphthylmethyl) penta a ]pyrido 3, 4 -bI indole, hydrochloride starch dried 250 mg 200 mg 55.0 43.0 magnesium stearate 460 mg 100. 0 The above ingredients are mixed and filled into haird gelati-n capsules in 460 mg quantities.

WO 95/24200 W095/4200PCTIUS95/03099 175 Formulation 12 Capsules each containing 20 made as follows: Amnount Per spiro-6, 6[2- 5-dimethoxy) 1,2,3, 4-tetrahydronaphthyl] 10-metI-1i-2,3,4,4a,5,6,7,llaoctahydr~.-H-indolo 3-c] quinuclidine, hydrochloride 20 mg mg of medicament are Concentration by Weight (ipercent) 10.0 44. 44.5 starch 89 mg microcrystalline cellulose mnagnesiumn stearate 89 mg 200 mg 100.0 The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 45 mesh U.S. sieve and filled into a hard gelatin capsule.

WO 95/24200 WO 9524200PCTUS95103099 176 Pormulation 13 Capsules each containing 100 mg of medicament are made as follows: Amount Per spiro-6, 6[2- (3-fluoro-4-methoxy) 1,2,3, 4-tetrahydronaphthyl] 10-methyl-2,3,4,4a,5, 6,7,llaoctahydro-1H-indolo guinuclidine, hydrochloride 100 mg by Weight 30.00 Dolyoxyethylene sorbitan monooleate starch powder 50 mg 0.02 250 mc 350 mg 100 .00 The above ingredients are thoroughly mixed and placed in an empty gelatin capsule.

WO 95/24200 PCT/US95/03099 177 Formulation 14 Tablets containing 10 mg of made as follows: Amount Per Tablet active ingredient are Concentration by Weight (percent) 8-fluoro-10-phenoxy- 6-(1-naphthylmethyl)- 2,3,4,4a,5,6,7,11coctahydro-lH-indolo- [2,3-c]quinoline, tartrate starch microcrystalline cellulose polyvinylpyrrolidone (as 10% solution in water) sodium carboxymethyl starch magnesium stearate talc 10 mg 45 mg 35 mg 4 mg 4.5 mg 0.5 mg 10.0 45.0 35.0 100 mg 100.0 The active ingredient, starch and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly.

The solution of polyvinylpyrrolidone is mixed wiht the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granule so produced is dried at 50°-600 C and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh U.S. sieve, are then added to the granule which, after mixing, is compressed on a tablet machine to yield a tablet weighing 100 mg.

WO 95/24200 PCT/US95/03099 178 Formulation A tablet formulation may be prepared using the ingredients below: Amount Per Tablet Concentration by Weight (percent) 8-methyl-10-methoxy- 6-(1-naphthylethyl)- 2,3,4,4a,5,6,7,11coctahydro-1H-indolo- [2,3-c]quinoline microcrystalline cellulose silicon dioxide fumed 250 mg 400 mg 10 mg 38.0 60.0 100.0 100.0 stearic acid 665 mg The components are blended and compressed to form tablets each weighing 665 mg.

WO 95/24200 PCT/US95/03099 179 Formulation 16 Suspensions each containing 5 mg of medicament per ml dose are as follows: per 5 ml of suspension 8-chloro-10-cyclopropyl- 6-(1-naphthylethyl)- 2,3,4,4a,5,6,7,llcoctahydro-lH-indolo- [2,3-c]quinoline 5 mg sodium carboxymethyl cellulose 50 mg syrup 1.25 ml benzoic acid solution 0.10 ml flavor q.v.

color q.v.

water q.s. to 5 ml The medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethylcellulose and syrup to form a smooth paste. The benzoic acid solution, flavor and color is diluted with some of the water and added to the paste with stirring. Sufficient water is then added to produce the required volume.

WO 95/24200 PCT/US95/03099 180 Formulation 17 An aerosol solution is prepared containing the following components: Concentration by Weiaht (percent) spiro-6,6[2-(3-ethyl-4-ethoxy)- 1,2,3,4-tetrahydronaphthyl]- 10-methyl-2,3,4,4a,5,6,7,llaoctahydro-lH-indolo[2,3-c]quinuclidine, maleate 0.25 ethanol 29.75 Propellant 22 (chlorodifluoromethane) 70,00 100.00 The active compound is mixed with ethanol and the mixture added to a portion of the Propellant 22, cooled to 0 C and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted further with the remaining amount of propellant. The valve units are then fitted to the container.

WO 95/24200 PCT/US95/03099 181 Formulation 18 A tablet formulation may be prepared using the ingredients below: Concentration Amount Per by Weight Tablet (Dercent) spiro-6,6[2-(3-ethyl-4-ethoxy)- 1,2,3,4-tetrahydro-6-methyl-naphthyl]- 10-methyl-2,3,4,4a,5,6,7,11aoctahydro-lH-indolo[2,3-c]quinuclidine, maleate 250 mg 38.0 microcrystalline cellulose 400 mg 60.0 silicon dioxide fumed 10 mg stearic acid 5 ma 665 mg 100.0 The components are blended and compressed to form tablets each weighing 665 mg.

Compounds of the present invention were tested for receptor affinity using the following procedure: IA. Biological Reagent Preparation.

Beef brain was removed immediately after slaughter, and choroid plexus were dissected over ice. Male Sprague-Dawley rats weighing 125-150 g (Harlan Industries, Cumberland, IN) were killed by decapitation. The brain of each was immediately removed and the cerebral cortex was dissected over ice. Tissues were hon 'genized in 9 volumes of 0.32 mol/L sucrose and centrifuged at 1,000 x g for minutes. The supernatant was centrifuged at 17,000 x g for minutes. The pellet was susperded in 100 volumes of 50 mM WO 95/24200 PCT/US95/03099 182 Tris-HC1 (pH7.4), incubated at 37 0 C for 10 minutes and centrifuged at 50,000 x g for 10 minutes, and the process was repeated three times. The final pellets were frozen at -70 0

C

and used within 2 weeks. Pellets were rehydrated with physiological buffer prior to use.

II. Assay Procedure.

Radioligand binding assays for 5-HTic and 5-HT 2 receptors were conducted according to described methods. The assays can be conducted as described by Hoyer D, Functional correlates of serotonin 5-HTI recognition sites, J. Receptor Res 8, 59-81 (1988) and Hoyer D, Engel G, Kalkman HO Molecular pharmacology of 5-HTI and 5-HT 2 recognition sites in rat and pig brain membranes: Radio-ligand binding studies with 3 H]5-HT, 3 H 8-OH-DPAT, 12 5 1] iodocyanopindolol, 3 H]mesulergine and [3H]ketanserin, Eur. J. Pharmacol. 118, 13-23 (1985).

For 5-HTIc receptor assays increasing concentrations of experimental compound, 50 mM Tris HC1 buffer pH 7.4 and tritiated mesulergine (2.0 nM) (3H ligand) were combined in polystyrene tubes at room temperature. The reaction was initiated by the addition of the resuspended choroid plexus tissue which had been preincubated at 37 0 C for 20 minutes. The reaction mixture was incubated in a 37 0 C water bath for 15 minutes.

The reactions were terminated by rapid filtration, (Brandel Cell Harvestor), through Whatman GF/B glass filters that had been presoaked in Tris buffer pH 7.4. The filters were then washed 2 times with 5 ml of ice cold Tris buffer pH 7.4. Washed filters were placed in scintillation vials and ml RedySolv, (Brandel), was added and samples were counted in a Searle D-300 beta counter. Means and standard error statistics were calculated for triplicate experimental determinations in certain cases. Mean values were obtained from three or more separate determinations. The incubation time for the reaction mixture was 15 minutes at 37 0

C.

WO 95/24200 PCT/US95/03099 183 Concentrations that caused a 50% inhibition of radioligand binding (ICso) and Hill coefficient were obtained by computer-assisted regression analysis.

Radioligand Binding Studies: Lembrane preparation from transformed cells.

Suspension cells expressing the cloned rat 5-HT2B receptor were harvested by centrifugation at 2,200 x g for 15 min at 4 0 C. Kursar, J. D. L. Nelson, D. B. Wainscott, M. L.

Cohen, and M. Baez, Mol. Pharmacol. 42: 549-557 (1992).

Membranes for the binding assays were prepared by vortexing the pellet in 50 mM Tris-HCl, pH 7.4 (0.5 x 109 cells/30 ml).

The tissue suspension was then centrifuged at 39,800 x g for min at 4 0 C. This procedure was repeated for a total of three washes, with a 10 minute incubation at 37 0 C between the first and second wash. The final pellet was homogenized in 67 mM Tris-HCl, pH 7.4 (at 20 40 and 12.5 million cells/ml, original cell number, for cells expressing low and relatively high levels of the 5-HT2B receptor, respectively) using a Tissumizer .(Tekmar, Cincinnati, OH), setting 65 for seconds.

3 H]5-HT binding studies. Binding assays were automated using a Biomek 1000 (.Beckman Instruments, Fullerton, CA) and were performed in triplicate in 0.8 ml total volume. Membrane suspension, 200 p1, (0.04-0.27 mg protein) and 200 gl of drug dilution in water were added to 400 gl of 67 mM Tris-HCl, pH 7.4, containing 3 pargyline, CaCl2, and L-ascorbic acid. Final concentrations of pargyline, CaC12 and L-ascorbic acid were 10 pM, 3 mM and respectively. Tubes were incubated at 37 0 C for 15 min or at 0°C for 2 hours (binding equilibria were verified for both of these conditions), then rapidly filtered using a Brandel cell harvester (Model MB-48R; Brandel, Gaithersburg, MD) through Whatman GF/B filters which had been presoaked in polyethylenimine and precooled with ice-cold 50 mM Tris-HCl, pH 7.4. The filters were then washed rapidly four times with one ml ice-cold 50 mM Tris-HCl, pH 7.4. The WO 95/24200 PCT/US95/03099 184 amount of 3 H]5-HT trapped on the filters was determined by liquid scintillation spectrometry (Ready Frotein) and automated using a Biomek 1000 (Beckman Instruments, Fullerton, CA) and were performed in triplicate in 0.8 ml total volume. Membrane suspension, 200 p1, (0.04-0.27 mg proten) and 200 .l of drug dilution in water were added to 400 .1 of 6" mM Tris-HCl, pH 7.4, containing 3 pargyline, CaC12, and L-ascorbic acid. Final concentrations of pargyline, CaC12 and L-ascorbic acid were 10 pM, 3 mM and respectively. Tubes were incubated at 37 0 C for 15 min or at 0C for 2 hours (binding equilibria were verified for both of these conditions), then rapidly filtered using a Brandel cell harvester (Model MB-48R; Brandel, Gaithersburg, MD) through Whatman GF/B filters which had been presoaked in polyethylenimine and precooled with ice-cold 50 mM Tris-HCl, pH 7.4. The filters were then washed rapidly four times with one ml ice-cold 50 mM Tris-HCl, pH 7.4. The amount of 3 H]5-HT trapped on the filters was determined by liquid scintillation spectrometry (Ready Protein and Beckman) and determined for best fit to a one-site or a two-site binding model using a partial F-test. De Lean, A. A.

Hancock, and R. J. Lefkowitz, Mol. Pharmacol. 21: 5-16 (1981). The following equation was used for a one-site binding model, Bound Bmax[L] Kd+[L] where Bound amount of 3 H15-HT specifically bound, Bmax maximum number of binding sites, Kd equilibrium dissociation constant and L] free concentration of 3 H]5-HT, or a two-site binding model, Bmaxlx[LJ Bmax2x[L] Bound= Kdl+[L] Kd2+[L] where Bound amount of 3 H]5-HT specifically bound, Bmaxl maximum number of high affinity binding sites, Bmax2 maximum number of low affinity binding sites, Kdl equilibrium dissociation constant for the high affinity site, Kd2 equilibrium dissociation constant for the low affinity WO 95/24200 PCT/US95/03099 185 site and free concentration of 3 H]5-HT. The values from the competition assays, the binding parameters for the IP3 standard curve and the EC50 and Eimax values from the IP3 assays were determined by nonlinear regression analysis of four parameter logistic equations (Systat, Systat Inc, Evanston, IL). De an, A. A. Hancock, and R. J. Lefkowitz, Mol. dharmacol. 21: 5-16 (1981). The values were converted to Ki values using the Cheng-Prusoff equation. Cheng, and W. H. Prusoff, Biochem. Pharmacol.

22: 3099-3108 (1973).

Compounds of this invention were tested using substantially the procedure describe in the Radioligand assay supra. and are summarized in Table I infra. The values in Table I are expressed as Ki values calculated as described supra. The blank values in Table I indicate that the compound was not tested in the corresponding assay.

co WO 95/24200 WO 9524200PCT[US95/03099 lable- 1

STRUCTURE

Ki Rat Ki Human 5-HT2B 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A

NH

N

H AVG. 6.62 N' SEM 0.09 N =4

OH

N

OH'

~OH AVG. =6.28 O0- N SEM 0.91 N =4 AVG, 12.20 SEM 54 N~kHN= 3 0 AVG, Fe,87 SEM =0.55 N=3 W 0 9 f 124 20 PCTUS9503I99 Iab-i-Q-

STRUCTURE

187 IDBzells r3fffSexatomn Ki Human Ki Rat 5-HT2A 5-HlT2A Ki Rat 5-HT2B Ki Humnan 5.IIT2B AVG. 18.98 090 5.12 0.32 1.21 =5 193525 N =6

H

0oN 0

N.,

NAVG. =15.11 15.09 2.05 SEM 2.43 2.26 0.29 237733 N =6 =5 0 NE~ AVG, 24.49 SEM N =2 AVG. 35.13

SEM=?

N=I

WO) 95124200 PTr[US95O3099 188 Iable1 IT2B 1ells [R~lJerotonin Ki flat Ki Human STRUCTURE 5-HT2B 5-HT2B 1125flDoT Ki Human Ki Rat 5-rT2A, 5-HT2A

I

AVG.

SEM=?

N 1 H OH 0ONo

N,

A17G. =10.24 74.49 8.91 215403 SEM 5.05 6.34 0.64 CIS ISOMER L N =4 =3 =3

H'OH

0 N.

N,N

N AVG. 11.36 107,74 12.44 SEM 4.21 16.32 2.29 215047 ISOMER L N 4 3 3 H{ OH 0 N.

N' AVG. 8.61 15.80 2.39 SEM 4.21 3.03 0.26 215046 ISOMER U N 4 3 3 WO 95/24200 WO 9524200PCTIUS95/03099 Ilh1lJ 189 52BHT I rM16em in f 1251]j)!2__ Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A STRUCTURE

OH

AVG. =9.35 SEM =4.36 215404 CIS ISOMER U N =4 18.60 2.42 3 2.44 0.35 3 c 0

HN

0

'N

AVG.= 11.87 SEM =1.93 N= 2 32.71 0.84 2 WO 95/24200 WO 95/4200 CT1US95/03099 Iabl

STRUCTURE

190 IT2Bicellr3H]Emwtoin

L-IMA

1125111201 Ki Rat 5-HT2B Ki Human 5-HT2B Y4 Human 5-HT2A Ki Rat 5-HT2A

ORK

AVG. =4115.29 8-OH-DPAT SEM =311.55 HBr N=3 AVG. =5153.15 SEM N =1

F

0 N N K AVG. =3019.67 SEW HCI N=1

NH~

0 I AVG.= 1501.95 N. SEM=?9 HCI N= 1 WO 95/24200 WO 95/4200 CTIUS95/03099 191 :CablI

STRUCTURE

5-H2. Q -ell rmsertonin I-MI2A rl25111D01 Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A AVG. =862.41

SEW-?

N= 1 HC1 AVG. 936.34 SEW N= 1 HCI 0 HcI AVG. =1752.56

SEM-?

N= 1

OH

0/0 N AVG. 79.27 =215.15 6158.98 SEM 5.39 4.97 2084.19 N N6 2 2 WO 95/24200 WO 9524200PCTfUS95/03099

IMLI

STRUCTURE

&H-B Ceh [r.Ti1 n~rn-nlnin LMiIA J125fl1DOI Ki Rat 5-HT2B Ki Human 5-HT2B Ei Human 5-HT2A Ki Rat AVG. =0.00 SEM N =1 =4749.52 =2229.48 508.94 45.08 3 354.03 41.34 =-3 0N Nk N AVG =784.17 127.21 L~il SEM= 32.35 28.34 HCI N= 3 3 AVG. -256.63 5690.86 SEW 9.56 560.80 HC1 N 3 3 WO 95124200 WO 9524200PCT[US95/03099 193 raHlSErotonin Ki Rat Ki Human STRUCTURE 5-HT2B 5-HT2B

LMIA

U25111201 Ki Human 5-HT2A Ki Rat 5-HT2A

N-

H

AVG. =-1018.01 6028.85 SEM HCI N1= N_,,AVG. =1789.87 0.00 6ac SEM== HCI N= =1 1 WO 95/24200 WO 9524200PCT[US95/03099 194

L-MIA

[125flDOI.

Ki Human i~ Rat 5-HT2A 5-HT2A Ki Rat 5-HT2B Ki Human 5-HT2B STRUCTURE T 0 B r CH AVG.= 37.58 SEM =4.76 CHi N=3

NH

2 B r AVG. =11.34 33.67 14.22 SEM =2.24 2.01 2.36 Hc1 N=3 3 3

NH

2 Br AVG. =32.03 SEM =3.49 Hc1 N=4

NH

2

NI{O

OI 8H/ AVG.= 283.84 -c( 8 H SEM 13.48 OH N =3 WO 95/24200 PCT/US95/03099 TableI 195 g1IBAQ1h mFMwerotoni _LI2AD12 Ki Human Kj Rat 5-HT2A 5-HT2A Ki ]Rat STRUCTURE 5-HT2B Ki Human 5-HT2B O NH 2

($OH

O N~ff AVG. 10.47 =11.91 SEM 1.46 =1.09 HON =4 =3

HO

B- NHI AVG. 130.79 Br SEM 18.70 HCI N 4 NH2 AVG. =10.19 9.84 7.77 15.80 ISEM =1.99 2.33 0.59 1.90 HO N=6 =5 3=3

N

2

NH

AVG. 9.15 0 SEM 1.47 \HC1 N =3 WO 95/24200 WO 9524200PCT/US95/0309 0 196 iaIl

STRUCTUR~E

5-HIM Cells r~qH-nSerotoni fL-HI2A r125nMOT Ej Rat 5-HT2B Kj Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A

NH

2

NH

C '9H ODHAVG. 146.84 =127.84 =120.16 0 SEM 13.49 =15.96

H

2 N N =7 3 -1 N

O

OH

~NH

I~.AVG. ,,69.22 0 SEM =50.27 N= 4

NH

2 AVG. =39.28 HC1SEM =14.04 Br N= 4

NH

2

NH

AVG. =112.90 SEM =5.61 HCI N=3 WO 95/24200 WO 95/4200 CT1US95/03099 197 32abe

STRUCTURE

a UBAICel L-UII2A f 125flDOI P'MlSeroto-niu- Kj Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Kj Rat 5-HT2A

NH

2 Br

HCI

AVG. =474.74 SEM =80.54 N

NH

AVG. =60.96 SEM =16.54 HCI

NH

2

NH

AVG. =32.78 HC1 I SEM =4.99 N=3 AVG. =5.65 HCI SEM =0.55 F N 3 WO 95/24200 WO 9524200PCT/US95O3099 TableJ

STRUCTURE

198 2Bll EMiIA Ki Rat 5-HT2B Ki Humnan 5-HT2B Ki Riunan 5-H? 4 Ki Rat 5-HT2A

NI{

2

NH

cI HC1

AVG.=

SEM=

6.21 0.55 6 AVG. =40.64 SEM =4.45 N= 3 3 ~F AVG. =15.37 F SEM =1.67 F---a/N=3 AVG. 30.18 SEM =0.90 N= 3 HCI WO 95/24200 PTU9/39 PCT/US95/03099 199 Table-2

STRUCTURE

5-H1I2B C-ells9 r3fflSerotonin

&-MIA

rL-2s~gTIDOI Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A

OH

OXO

HO

AVG. =84.49 SEM =8.44 N= 3

-N

HOX

NH

0 AVG. 38.48 SEM= 3.77 IN 0

NH

2

CH-

AVG. 49.29 0 SEM N=1

NH

HcI AVG-. =6.52 SEM =0.60 N =2 WO 95/24200 PCT[US9503099 Table 1

STRUCTURE

200 r3Hiserotoin LULI2A r12 511O i Human Rat 5-HT2A 6-HT2A Ki Rat 5-HT2B Ki Human 5-HT2B

NH

2 NH AVG. 49.92 SEM 11.09 F N=2

NH

2

NH

HO\/

HO AVG.= 41.62 creatinine SEM 10.13 sulfate salt N 2

NH

2

NH

HO OH AVG. 4571.89 creatinine SEM 499.67 sulfatesalt N 2

NH

2

NH

AVG. 154.84 \Hc N=1

I

WO 95/24200 WO 9524200PCT1U595/03099 Table I 2~ [311]Serotonin_ Ki Rat Ki Humian STRUCTURE 5-HT2B 5-JJT2B L M2 A Ki Human 5-HT2A Ki Rat 5-HT2A AVG. =20.85 SEM =5.29 N= 2 NH2 F AVG-'. 62.43 F SEM 7.52 HCI N =3

NH

AVG. =102.06 SEM 1.62 2 INH o AVG. =14.78 SEM N=1 'WO 95124200 WO 9524200PCT1US95/03099 202 r3nlbgerotonijn Ki Rat Ki Human STRUCTURE 5-HT2B 5-HT2B 51iI2A 121 of Ki Human 5-HrD2A Ki Rat 5-HT2A

NH

2 "c AVG. =7.94 11.21 27.55 =20,7q NI SEM 0.52 =3.48 01 N 6 =1 3 =3 0 NH 1 F

Q

\NH AVG. =336.55 SEM F N=1 WO 95/24200 WO 9524200PCT/US95/03099 203 Iablel

STRUCTURE

Nr ff Npratomi 5-HT2 rl25n1DOI Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A

(N

'N

HCI

AVG. ='4.50 SEM 0.47 N 12 3.79 0.80 8 0 s- N

I.

0 AVG. =3.58 SEM =1.67 N 3 WO 95/24200 PCTIUS9S03099 204 Iabk-

STRUCTURE

F3flserotnin I-HaIA [12511DOT Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-Hr 112A Ki Rtat 5-HT2A WC AVG.= 3.17 21.74 15.74 Br SEM 0.36 0.74 1.41 HCl N=3 3 3

NH

O0 SAVG. 34.49 0- SEM 2.85 0-H N=4 0' AVG. 5.61 1.40 44.46 OH ,0 0 SEM 0.91 0.08 0.75 ISOMERA N=5 5 3

HO'

0 0 0' AVG. 30.07 5.55 372.81 OH -SEM 8.51 0.40 23.51 ISOMER B N 5 4 3 -i Y I WO 95/24200 WO 9524200PCT[US95/03099 205 I-2ZBJfllls 5-HT2A F 12511DOT Ki Rat 5-HT2iB Ki Human 5-HT2B Ki Hum.An 5-11l' A Ki Rat.

5-HT2A STRUCTURE 110, AVG. =8.16 1.50 SEM =2.16 0.35 N= 4 4 23.39 3.81 3

HCI

AVG. =3.10 0.79 SEM =0.20 0.06 N=3 7 28.07 2.30 3

NH

1 SN=0.68 HC N N3 0

XN

00,HAVG. =28.37 K~0H0- SEM =2.08 0 N=3 WO 95/24200 WO 9524200PCT/US95/03099 iabi&-J

STRUCTURE

206 2:B Cells r3HlSerotonin Adfl2A r125flDOI Ki Rat 5-HT2B JKI Human 5-HT2B Ki Human 5-HT2A Ki Rat

NH

SOH '/AVG. =3.94 1.29 5.62 OH /SEM =0.88 0.12 0.63 O N=3 =33 N NH ll~: AVG. =39.70 SEM =5.09 HCI o N=3 AVG. =1463.01 SEM =110.95 N 4 0

,'OH

KOH

0 AVG.= 14.18 SEM =1.74 N 4 WO 95,24200 WO 9524200PCT/US95IO3099

ILII

STRUCTURE

207 5-~i2 Cella I3Hlberotnin Ki Human Ki Rat 5-HT2A 5-HT2A Kj Rat 5-HT2B Ki Him an 5-HT2B 0 AVG. =44.91 SEM 1.48 N= 3 AVG. =5.02 SEM =0.51 N= 3 1.64 0.23 4 0.83 0.10 3 2 HC1 H 2 0 0

OH

0 AVG. =4.82 SEM =0.23 N= 3 NN NH AG. =2.16 0' SEM= 0.33 HC1 N 3 WO 95/24200 WO 9524200PCTIUS95/03099 yzhb&-i

STRUCTURE

208 ~U2BJc1lls r[mslrtonin R125fl

OT

Ki Rat 5-HT2B Ki Human 5--TT2B Ki Human 5-HT2A R Rat 5-HT2pA

NNIH

AVG. =228.87 0SEM =18.34 HCI N= 3 H

NHI

OH

OH 0 AVG.= 6.42 =20 0 -1 SEM= 0.78 =-9 OH N=3 =1

F

N

NH

~OH AVG.= 4.38 SEM 1.25 o N=3

NH

3.31 0.74 4.63 HCI SEM =0.31 0.04 0.26 N=3 =3 3 wo 95/24200 WO 9524200PCT/US9SIO3099 209 5-2 BtClls F3fflSerotopin r125flDOI Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HTA Ki Rat 5-HT2A STRUCTURE

AVG.=

SEM=

N=

219.79 20.68 3 P .AVG. =4609.36 SEM =316.40 N= 3 HCI AVG. =379.15 SEM =16.72 N= 3

HCI

AVG. =114.16 SEM =7.17 N= 3 WO 95/24200 WO 9S24200PCT/US9SIO3O99 lable I

STRUCTURE

2 5-HT A'R ell [3111 Sertonin 5 2 A f1251nDOT Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A AVG. 404.85 SEM 51.64 N= 3

AVG.

SEM-

N-

97.53 AVG. =71.75 SEM N= 1 AVG. =70.71 SEM =10.08 N= 3

HCI

WO 95/24200 WO 95/4200 CTfLJS95/03099 Table 1

STRUCTURE

211

LEEBQPIE

[3nlserotonin ~flI2A [1251IDOI Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A AVG. =15.83 SEM =1.73 N=3 HCI

NH

AVG. =13.98 16.43 41.71 47.00 SEM =1.00 7.42 5.15 HC1 Br N=3 1 4 3

NH

NH'

/AVG. 6.20 9.68 22.72 25.61 Br SEM 0.62 1.12 2.15 3.43 HCI N =3 3 6 3

NH

~NH

HCI AVG. =62.09 SEM =1,76 Br N 3 WO 95/24200 PTU9139 PCTIUS95/03099 lable-

STRUCTURE

212 Ea3llerotonip-

LIMA

r1 Ki Rat 5-HT2B Ki Human 5-HT2B Kj Human 5-UiT2A Ki Rat 5-HT2A F AVG. =57.89 FF SEM =7.60 HCI F N3 AVG. =32.44 y SEM =3.48 Br HCI N=3 AVG. =322.26 SEM =35.50 N=3 F HC1

NH

\/NH AVG.= 25.63 F SEM 2.59 Fc N =3 WO 95124200 WO 9514200 PCTIUS95/03099 lable.1

STRUCTURE

213 L-M2A r12511D01) Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HT2A Ki Rat 5-HT2A AVG. =78.46 SEM =1.22 N= 2

HCI

HII~

cl

HCI

AVG. =60.10 SEM =2.07 N= 3 AVG. =2.69 2.68 20.61 15.49 SEM =0.24 0.24 1.28 0.79 N= 6 =4 4 =3 Cf--

N

Eaci I' AVG.= 3.19 0- SEM =0.33 N= 3 0.84 0.15 4 0.93 0.07 3 WO 95/24200 WO 9524200PCIVVS95/03099 214 Tablel

STRUCTURE

5-2TB CI1 rqmn~qrnfnin

L-MA

F 125flDOI Ki Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HrD2A Ki Rat 5-HT2A AVG. =8.77 SEW N= 1 P- \NH 0 AVG. =5652.17 6z ,SEM HCI N= 1

N

I AVG. =15.26 10.15 17.76 I SEM=? 2.19 0.68 HCII N= 1 =4 3

N'

H N

H

OH

0 AVG. ==1.69 49.70 0 o- SEM OH 0\N= =1 1 WO 95/24200 WO %I42O(~PCr/US95/03099 215 Tabl

STRUCTURE

X2B Clls r3Tqllserotonin EMiIA

K

1 Rat 5-HT2B Ki Human 5-HT2B Ki Human 5-HrP2A Ki Rat

NFH

H

O H

OH

AVG.=

SELM=

-2.08 =6.71

AVG.=

SEM=

-0.37 -1 =12.23 =1 HC1

OH

OH

0,AVG.

0\

SEM=

-0.87 =17.48 -9 N

N

HAVG.= 1.47 54.85 H I0 SE M? HC1 HO N=1 WO 95/24200 WO 9524200PCTIUJS95iO3O99 216 Table I 5-HT2BII [3HjSertonin p Ki Rat Iii Hiuman SIRUCTURE 5-HT2B 5-HT2B Ki Human Kj Rat 5-HT2A 5-HT2A

AVG.=

SEM=

=3.46 -1 -201.26 WO 95/24200 WO 9524200PCTUS95IO3O99 Tab-Je

STRUCTURLE

217 I2I B Cll [3H]Serot2in_ r1T2 Ki Rat 5-HT2B Ki Human 5-HT2B Y4 Human 5-HT2A Ki Rat 5-LIT2A N{-aOH AVG. =44.65 SEM =25.15 N=5 12.44 1.70 3 1.25 0.56 5 5.36 1.85 255747, Trans HN-aOH AVG. 22.29 SEM 6.65 N =5 =4.30 S0.30 =3 11.33 3.26 =-5 1.57 0.49 278458, C4IS

AVG.

SEM

N

-30.90 -4.52 -3 6.24 0.29 3 21.18 3.78 3 2.82 0.40 3 253535 (TRANS) WO 95/24200 WO 9524200PCT[US95/03099 Table

STRUCTURE

218 [,3H Serotonin L12 OLLQ Ki Rat 5- Flr'2B Ki Human 5-HT2B Kj Human 5-HT2A Ki Rat 5-HT2A

OH

AVG.=

SEM=

-7.87 =0.60 =1

OH

AVG.=

SEM=

=2.24 =21.92 I1 WO 95/24200 PCT/US95/03099 219 The following Cell assays use Human Cells Comoound 5HT Cells 5HT Cells 5HT2 CelIs Example 100 16.44 292.58 351.96 Example 105 22.07 86.48 195.44 Example 102 268.49 917.16 2172.86 Example 106 367.41 263.94 1108.87 Example 104 11.35 32.99 52.06 Example 97 9.56 123.93 220.51 Example 99 106.17 556.40 1117.00 Example 107 177.89 362.79 325.10 Isomer 107(1) 142.80 152.65 137.76 Isomer 107(2) 2894.33 1967.05 6211.80 Example 101 121.19 172.03 783.35 Example 98 52.54 53.65 202.60 Example 96 667.82 277.62 976.73 Example 95 839.63 3443.51 2641.21 Example 103 3520.31 1447.55 9247.06 Assay Methods 5-HT2B receptor in tissue in vitro: Male Wistar rats (150-375 g; Laboratory Supply, Indianapolis, IN) were sacrificed by cervical dislocation, and longitudinal section of the stomach fundus were prepared for in vitro examination. Four preparations were obtaine from one rat fundus. Cohen, M.L. and J. Pharmacol. Exp.

Ther. 233:75-79 (1985). Tissues were mounted in organ baths containing 10 mL of modified Krebs' solution of the following composition (millimolar concentrations): NaCl, 118.2, KCI, 4.6; CaC1 2 -H20, 1.6; KH 2

PO

4 1.2; MgSO4, 1.2; dextrose, 10.0; and NaHCO3, 24.8. Tissue bath solutions were maintained at 37 0 C and equilibrated with 95% 02 and 5% CO 2 Tissues were placed under optimum resting force (4 g) and were allowed to equilibrate for approximately 1 hour before exposure to the test compound. Isometric contractions were recorded as changes in grams of force on a Beckman Dynograph with Statham UC-3 transducers.

~_aY I WO 95/24200 PCT/US95/03099 220 Determination of Apparent Antagonist Dissociation Constant: Noncumulative contractile concentration-response curves for serotonin and other agonists in the fundus were obtained by a stepwise increase in concentration after washing out the.preceding concentrations every 15-20 minutes.

Each agonist concentration remained in contact with the tissue for approximately 2 minutes and maximum response to each compound concentration was measured ED50 values were taken as the concentration of agonist that produced halfmaximal contraction. After control responses were obtained, tissues were incubated with an appropriate concentration of buffer or antagonist for 1 hour. Responses to serotonin were then repeated in the presence of an antagonist.

Concentration responses utilized only one agonist and one antagonist concentration per tissue. In general, successive agonist responses in the presence of buffer treatment were unaltered (average dose ratio was 1.28 0.21).

Apparent antagonist dissociation constants (KB) were determined for each concentration of antagonist according to the following equation: KB= [B]/(dose ratio-1) where is the concentration of the antagonist and dose ratio is the ED 50 of the agonist in the presence of the antagonist divided by the control ED50. Generally, parallel shifts in the concentration-response curves occurred in the presence of antagonists. The results were expressed as the negative logarithm of the KB -log KB). Calculations were completed using known methods.

The results of the in vitro assay for certain compounds of this invention are presented in Table II. The values in Table II are expressed as -log Kg standard error (number of data points). The 5 -HT2B value represents the negative log of the concentration of an antagonist that will produce a two fold dextral shift in the concentration response curve to serotonin in the rat stomach fundus which is mediated by 5-HT2B receptors. Likewise, the 5-HT2A value -rs I~ -s~ R ~1I~II_ WO 95/24200 PCTUS9503099 221 represents the negative log of the concentration of an antagonist that will produce a two fold dextral shift in the concentration response curve to serotonin in the rat jugular vein which is mediated by 5 -HT2A receptors. The blank values in Table II indicate that the compouna was not tested in the indicated assay.

i I -N WO 95/24200 WO 9524200PCTIUS95/03099 222 _____TableI 11_ Example 5 -HT2B 5 -HT2A (Fundus) (jugu~lar) 1 9 .00 0 0.7 (3) 2 3 8.78 0 0.24 (4) 4 8.92 ±0.29 (4) 6 9.60 ±0.13 (7) 7 9.02 ±0.35 (3) 8 8.45 0.24 9 9.30 ±0.12 (7) 9.22 0.05 11 <7.52 12 9.29 ±0.18 13 8.50 ±0.13 (4) 14 9.61 0.22 9.34 ±0.12 16 9.71 ±0.14 8.15 0.28 (3) 17 9.46 ±0.11 7.66 0.13 (4) 18 8.80 ±0.17 19 10.12 ±0.18 (3) 9 .48 0.3 0 7 .21 0 .2 0 (4) 21 22 8.21 ±0.43 (3) 23 24 26__ 27 28 8.55 ±0.10 (4) 29 8.12 ±0.16 (7) 8.89 ±0.12 (4) 31 8.95 ±0.17 7.29 ±0.09 (4) 32 WO 95/24200 PCT/US95/03099 223 TableII_(cont.) Example 5-HT2B 5 -HT2A (Fundus) (jugular) 33 34 9.42 ±0.18 *9.06 ±0.2,7 (3) 36 9.80 ±0.15 8.14 0.10 (6) 37 9.19 ±0.14 38 39 8.32 ±0.17 9.75 ±0.11 41 9.81 ±0.18 7.94 ±0.15 (6) 42 9.56 ±0.22 (3) 43 9.44 ±0.16 (6) 44 8.40 ±0.40 (3) 8.14 ±0.32 (3) 46 9.37 ±0.11 8.22 0.07 (12) 47 48 49. *10.41 0.22 8.40 ±0.28 (3) 51 9.75 ±0.11 8.07 ±0.10 (8) 52 *9.10 ±0.28 (3) 53 54__ 8.95 ±0.07 (4) 56 *7.53 ±1.08 57 <8.0 (3) 58 <7.52 (4) 59 9.69 ±0.21 8.92 ±0.04 61 8.44 ±0.22 62, 8.58 ±0.23 63 9.09 ±0.23 64 ).73 ±0.05 Approximate value WO 95/24200 PCT/US95/03099 224 Non-competitive inhibitors at 30 nM Functional In vitro assay: Sprague-Dawley rats (200-250g; Laboratory Supply, Indianapolis, IN) were sacrificed by cervical dislocation and 8 cm segment of distal colon was removed and washed in ice cold modified Kreb's solution of the following composition (millimolar): Nacl, 118.2; Kcl, 4.6; Cacl2. H20, 1.6; KH2PO4, 1.2; MgSO4, 1.2; dextrose, 10.0; and NaHCO3, 24.8.

The colon was mounted on a glass rod and the longitudinal muscle layer with attached myenteric plexi was removed and mounted in organ baths, containing above described Kreb's solution maintained at 37 0 C and equilibrated with 95% 02 and CO2. Tissues were placed under 2 g tension and allowed to stabilize for 1 hour. Isometric contractions were recorded as changes in grams of force using grass FT03 transducers and

MI

2 computerized dynograph system. Cumulative concentration-response curves for serotonin were obtained by a stepwise increase in concentration after washing out the preceding concentration for 10-15 minutes. Each agonist concentration remained in contact with the tissue for minutes. Maximum response to each concentration was d'termined and digitized. EC50 values were taken as the concentration of agonist that produced half maximal contraction. After control responses were obtained, tissues were incubated with an appropriate concentration of antagonist for 15 minutes. Response to serotonin were then repeated in the presence of an antagonist. Concentrationresponse utilized only one concentration of antagonist per tissue. Apparent antagonist dissociation constants (KB) were determined for each concentration of antagonist according to the following equation: KB= [B]/(dose ratio where is the concentration of the antagonist and dose ratio is the ED50 of the agonist in the presence of antagonist divided by the control ED50. The

I--

WO 95/24200 PCT/US95/03099 225 results were expressed as the negative logarithm of the KB (i.e.,-log KB) (Br. J. Pharmacol. Methods 4:4165, (1980).

The functional in vitro method described supra. was used to test compounds of this invention. Results obtained using the functional in vitro assay are presented in Table III. Values are expressed as pKi and pA2 (-log KB). The following table illustrates the results obtained when the compounds were tested using the Radioligand assay supra.

(pKi) and the functional in vitro method described supra.

(PA2) Table III Compound Example 73 Example 49 Example 20 Example 72 Example 41 Example 17 Example 22 7-methyl-8-chloro-l,2,3,4tetrahydro-9H-pyrido[3,4b]indole 6-bromo-l,2,3,4-tetrahydro- 9H-pyrido[3,4b]-indole 6-chloro-l,2,3,4-tetrahydro- 9H-pyrido[3,4b]-indole nKi nA i-fLLL--- lin 7.85 8.4 8.51 7.8 8.19 8.09 8.27 8.57 7.21 7.15 8.9 8.2 7.8 7.2 6.2 4.8 8.3 8.2 7.2 In vivo Studies: Sprague-Dawley Rate (250-300 g) were fasted overnight. The rats were anesthetized with urethane (250 mg) delivered intraperitoneally. The abdominal cavity was opened and strain guage transducers were sewn on the antimesenteric border of the colon. The transducers were oriented to record circular muscle contractions. The animal body temperature was maintained by a heating pad. An intravenous catheter was inserted into the jugular vein for drug administration. The carotid blood pressure was also monitored. Output of the i- II--- WO 95/24200 PCTIUS95/03099 226 strain guage transducers was graphed on a Beckman Dynograph.

Baseline motility was monitored for 30 minutes. At the end of the 30 minute period, a vehicle control dose was administered and motility was recorded for an additional minutes. A serotonin dose -esponse was developed.

Successively higher doses of serotonin were administered at minute intervals. An ED 50 dose was calculated, which was the dose producing half maximal contraction. In antagonist experiments, historical ED 50 dose was administered to validate the experimental set up. Next, a dose of antagonist was given. The motility was monitored for 15 minutes. After the minute monitoring, an ED50 dose was administered. Motility was evaluated by measuring the number of contractions and multiplying them by the amplitude of contractions over a set time period to provide a Motility Index. The percent inhibition was calculated from the vehicle (no antagonist) treated group. A minimum of three rats were used for each concentration and data from different animals was pooled to determine ED 50 values.

Compounds Of this invention proved to be active using the in vivo method described supra. For example, the compound of Example 73 produced an ED 50 value of 3.2 rag/kg, i.v.

D-

Claims (2)

1. 9. 9 9 9**9 .9 9 9 9* 9 99**99 4 9 9* 9. *999 9, 00 90 009909 4 0090 9 0 9000 9 900000 l0 wherein: R, 5 is C,-C 4 alkyl; R 16 is allyl or CX- 4 straight chain alkyl; is hydrogen or CX- 4 straight chain alkyl; R 1 8 is hydrogen, C 1 -C 4 alkyl, hydroxy, or (J 1 -C 4 alkyloxy; in'is 0, 1, 2, or 3; a compound of Formula V N:IlibcO 1608 wherein: R 19 1S CI-C 4 alkyl; R 20 is allyl or C 1 -C 4 straight chain alkyl; R, 1 is hydrogen or CX- 4 straight. chain alkyl; R, 2 is pyridinyl or imidazolyl; alk is a divalent organic radical derived from a straight or branched C 1 -C 5 alkane; a comnpound of Formula VI 0 0 R 2 N ci:)3 N' IV wherein: R 2 3 is C 1 -C 3 alkyl or allyl; R 24 is C 1 -C 3 hydroxyalkyl or CI-C 3 dilydroxyalkyl; R 2 is hydrogen or CH 3 a compound of Formula VII H H 0IbOI0 *~CH 3 VII a compound of Formula ViII of. I..I ix whren yb ncmiainwt4h abnatmt hc ti ond eie aru coonditn of ruaI WAbU01608 'R29 R 2 9 R 26 is hydrogen, C 1 .Calkyl, allyl, or ;R 27 is hydrogen, C 1 -C~AIclk, allyl, Nor (CH 2 n' is 1 to 5; XV is an optionally substitte phenyl, C,- C~alkoxy,, -rl C 1 -C 3 alkylthio; R 2 8 and R 2 9 afe independently hydrogen, Cl-C 3 alky',, C 1 C 3 alkoxy, hydroxy, C,-C~alkylthio, halo, CN, phenyl; or together are (CH 2 P" is 3 to 6; Y" is CR 2 0, in" is 0, 1, or 2; and a compound of the Formula X \N N 0 NH I V a *09 or a pharrnaceutically acceptable salt or solvate thereof. 2. A method of Claim 1 wherein the compound is selected from the group consisting of Formula 1, 11, and IX. A method of Claim 2 wherein thle comipound of Formula IX has the following structure: 0 R 29 R 26 wherein R1 6 R 27 R, 8 R 19 and Ya are as defined supra. 4. A method of Claim 2 wherein the compound is selected from the group consisting of 2..(di-n-propylainino)-8-(isothiazol-3-yl)-1 ,2,3 ,4-tetrahiydronaphthalene, 2- ethylamino-8'(isoxazol-3-yl)- 1,2,3 ,4-tetrahlydronaphthalene, 2-(N-methyl-N-benzyl 1 ,2,3-oxadiazol-4-yl)- 1,2,3 ,4-tetrahydronap~hthalene, 2-dial lyl 2o amrino-8-(pyrazol-3-yl)- 1,2,3, 4-tetrahydronaphthalenie, 2-diethylamino-8-( 1,3 ,4-oxadiazol- 2-yl)-1 4-tetrahydronaplhthalene, 2-(di-n-propylamino)-8-(3-methioxypyrid-2-yl)- 1,2, ,4-tetrahiydronaphthialene, 2-benzylmethylaniiino-8-(3-methoxypyrid-2-yl)- 1,2,3 ,4-tetra Shydronaphithalene, 2-benzylmethylamino-8-(benzofuiran-2-yl)- 1,2,3 ,4-tetrahiydro N.A1ibcO I 6OB 228 naphthalene, 2-dimethylarnino-8-(1 5-triazin-2-yl>- 1,2,3 ,4-tetrahydronaphthalene, 2- (dicyclopropylmethylamino)-8-(oxazol-4-yl)- 1,2,3, 4-tetrahiydronaphthalene, 2-ethylam-ino- 1,2, 3-oxadiazol-4-yl)-thio- 1,2,3, 4-tetrahydronaphthalene, 2-n-butylamino-8-(5- m-ethoxypyrimidin-2-yl)- 1,2,3, 4-tetrahydronaphthalene, 2-(di-n-propylamino)-8-(5- chlorooxazol-2-yl)- 1,2,3, 4-tetrahydronaphthalene, 2-(di-n-propylamino)-8-(pyrimidin-2- yl)-l ,2 ,3 ,4-tetrahydronaphthalene, 2-(di-n-propylai-nino)-8-(2-aminopyrimiidin-4-yl)- 1,2, 3 ,4-tetrahydronaphthalene, 2-(di-n-propylamino)-8-(3-phenyl- 1,2 ,4-oxadiazol-5-yl)- I,2, 3, 4-tetrahiydronaphithalene, 2-(di-n-propylarnino)-8-(3-rnethyl- 1,2,4-oxadiazol-5-yi)- 1,2,3,4- tetrahydronaphthalene, 2-(di-n-propylamino)-8-(pyrazin-2- yl)-l 4-tetrahydro naphthalene, 2-(di-n-propylamiino)-6-(broniopyrazin-2-yl)- 1,2,3, 4-te-trahydronaphthalene, 2-(di-n-propylamino)-8-(benzothiazol-2-yl)- 1,2,3, 4-tetrahydronaphthalene, 2-(di-ii-propyl amino)-8-(benzoxazol-2-yl)-1 3,4-tetrahy!drona-phthalene, 2-(di-n-propylamino)-8-(indol- 3-yl)-l ,2 ,3,4 -tetrahydronaphithalene, 3-(di-n-pronylamino)-5-(isoxazol-2-yl)- 1,2,3 ,4-tetra to 0 0 0 hydronaphthalene, 3-(di-n-propylaimino)-5-(isoxazol-2-yl)-chromane, 5-(isoxazol-5-yl)-3- :0is 1 (dipropylar.-ino)chromane, 5-(3-methylisoxazol-5-yl)-3-(dipropylamino)chromane, 5-(4- methylisoxazol-5-yl)-3-(dipropylaminio)chromalie, 4-dimethiylisoxazol-5-yl)-3- (dipropylarnino)chromane, 5-(3-methylisoxazol-5-yl)-3-(dipropylamino)thiochromane, ~.:(4-m-ethylisoxa,1r4l-5-yl)-3-(Iipropylamino)thiochromane, 4-dimethylisoxazol-5-yl)-3- *(dipropylarnino)thio!.tromnane, and 8-(4 67-tetrahydrobenz[cisoxazol- 1-yl)-2-(dimiethyl amino)tetrahydronaphthalene.; or a pharmaceutical salt or solvate thereof. A method of Claim 2 wherein the compound is a compound of Formula I or 0000 6. A ethod of Claim 5 wherein the compound is selected from the group consisting of 7-bromo-8-miethyl- 1,2,3, 4-tetrahydro-9H-pyrido[3 ,4b]-indole, 6-isopropyl- ol. 25 8-methoxy- 1,2,3, 4-tetrahydro-9H-pyrido[3 ,4b]-inidole, 5-chloro-8-ethoxy- 1,2,3, 4-tetra hydro-9H-pyrido[3 ,4b]-indole, 6-chloro-7-methiyl-8-flutoro- 1,2,3, 4-tetrahiydro-9H-pyrido [3 ,4b]-indole, 5-dim-ethiylamino-8-hyclroxy-i ,2 ,3,4-tetrahiydro-9H-pyrido[3 ,4b]-indole, 6- nitro-8-butyl- 1,2 ,3,4-tetrahydro-91--pyrido[3 ,4b]-indole, 7-cyclohexyl-8-hydroxy- 1,2,3,4- tetrahydro-9H-pyrid'o[3 ,4b]-indole, 6-[3-miethiyl-cyclohexyl]-8-methiyl- 1,2,3 ,4-tetrahydro-
2. 911-pyrido[3 ,4b]-indole, 6-benzyl-8-fluloro- 1,2,3, 4-tetrahiydro-9H-pyrido[3,4b] -inidole, cyclohlexylmethyl-8-chloro- 1,2,3, 4-tetrahiydrc-91--pyrido 4b]-inidole, 6-carboxyl-8- bromno- 1,2,3, 4-tetrahydro-9H-pyrido[3 ,4b]-indole, 6-ethioxy- 8-isopropyl-3-mlethyl- 1,2,3, 4-tetrahiydro-9H-pyrido[3 ,4b]-indole, 6, 8-dichloro-4-naphthylmnethiyl- 1,2,3 ,4-tetrahlydro- 9}{-pyrido[3 ,4b]-indole, 6, 8-dim-ethiyl-3 ,4-dimethyl- 1,2,3 ,4-tetrahvdro-9HWpyrido[3 ,4b}- indole, 7, 8-difluoro-2-(N)-methyl- 1,2,3, 4-tetrahydro-9H--pyrido [3 ,4b]-indole, 6, 8-dibu~tyl- 2-(N)-cyclopropylmethiyl- 1,2,3, 4-tetrahydro-9H-pyrido[3 ,4b]-indole, 6, 8-dibromo-2-(N)- cyclohexenylmethyl-1 ,2,3 ,4-tetrahydro-9H-pyrido[3, 4b]-indole, 8-chloro-2-(N)-benzyl- 1 ,2,3,4-tetrallydro-9H-pyrido[3 ,4b]-indole, 8-fluloro-4-methlyl-2-(N)-cyctlhexyl-1,2,3,4- Stetr-ahydro-9H--py riclo 4b]-indole. 6-miethylainie-8-chlloro-3-isopropyl- 1,2,3, 4-tetra N:AWbcO 1608 hydro-9H-pyrido[3,4b]-indole, and 6-chloromethyl-8-chloro-1,2,3,4-tetrahydro-9H-pyrido [3,4b]-indole; or a pharmaceutical salt or solvate thereof. 7. A method as claimed in any one of Claims 1 to 6 wherein the compound is selected from the group consisting of Formula III, IV, V, and VI. s 8. A method for blocking a 5HT 2 receptor in a mammal, comprising administering a 5HT 2 B receptor blocking dose of a compound selected from the group consisting of Formula I, II, III, IV, V, VI, VII, VIII, IX, and X supra.; or a pharmaceutically acceptable salt or solvate thereof. 9. A method for selectively blocking a 5-HT 2 B receptor in a mammal, comprising administering a 5-HT 2 selective compound selected from the group consisting of Formula I, II, III, IV, V, VI, VII, VIII, and IX supra.; or a pharmaceutically acceptable salt or solvate thereof to a mammal. A method for blocking a human 5-HT2B receptor in a human, comprising administering a 5-HT 2 B blocking dose of a compound selected from the group consisting 15 of Formula I, II, III, IV, V, VI, VII, VIII, and IX supra.; or a pharmaceutically acceptable salt or solvate thereof to a human. Dated 19 September, 1996 Eli Lilly and Company 0 .a Patent Attorneys for the Applicant/Nominated Person 20 SPRUSON FERGUSON N:/libc/01608 -J Ic~CIBd~ INTERNATIONAL SEARCH REPORT ln..rnational application No. PCT/US95/03099 A. CLASSIFICATION OF SUBJECT MATTER IPC(6) A61K 31/55 US CL :514/220,255,285,288,292,339 According to Internaticral Patent Classification (IPC) or to both national classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) U.S. 514/220,255,5,28,288,292,339 Documentation searched other than minimum documentation to the extent that such documents are included inthe fields searched Electronic data base consulted during the international search (name of data base and, where practicable, search terms used) C. DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. A US, A, 4,520,024 (Cohen) 28 MAY 1985, see entire 1-42 document. Further documents art listed in the continuation of Box C. See patent family annex. Special categories of cited documents: documentdefinig the general state of the art which is not considered to be of particular relevance earlier document publihed on or after the international filing date document which may throw doubts on priority claim(s) or which is cited to establish the publication date of another citation or other speial reason (as specified) document referring to an oral disclosure, use, exhibition or other means document published prior to the internationl filing date but later than the nrioritv date claimed "T later document published after the intemational filing dole or priority date and not in conflict with the application but cited to understand the principle or theory underlying the invention document of particular relevance; the claimed invention cannot be considered novel or caonot be considered to involve an inventive step when the document ia taken alone document of particular relevance; the claimed invention cannot be considered to involve an inventive step when the document is combined with one or more other such documents, such combination being obvious to a person skilled in the art document member of the same patent family Date of the actual completion of the international search Date of 18 JUNE 1995 Name and mailing address of the ISA/US Commissioner of Patents and Trademarks Box PCT Washington, D.C. 20231 Facsimile No. (703) 305-3230 Form PCT/ISA/210 (second sheet)(July 1992)* i