CA2176127A1 - Methods for the treatment or prevention of conditions associated with amyloidogenic reptides - Google Patents

Abstract

This invention provides methods for the treatment or prevention of physiological disorders associated with an amy loidogenic peptide by the administration of a series of substituted naphthalenes, dihydronaphthalenes, benzofurans, benzothiophenes indoles, or the pharmaceutically acceptable salts or solvates thereof. This invention also provides methods for the treatment or prevention of Alzheimer's Disease or Down's Syndrome by the administration of these same benzofurans or benzothiophenes or the pharmaceutically acceptable salts thereof.

Description

Wog5/l7os~ - 1 - PCT~S94/1~655 METHODS FOR THE TREATMENT OR PREVENTION OF
CONDITIONS ASSOCIATED WITH AMYLOIDOGENIC PEPTIDES
.

Alzheimer's disease is a degenerative disorder of t the human brain. Clinically, it appears as a progressive dementia. Its histopathology is characterized by degeneration of neurons, gliosis, and the abnormal deposition of proteins in the brain. Proteinaceous deposits (called 10 "amyloid" ) appear as neurofibrillary tangles, am,vloid plaque cores, and amyloid of the congophilic angiopathy. [For a review, see, D.J. Selkoe, Ne~lron~ 6:487-498 (1991)~
While there is no general agreement as to the chemical nature of neurofibrillary tangles, the major constituent of both the amyloid plaque cores and the amyloid o~ the congophilic angiopathy has been shown to be a 4500 Dalton protein originally termed ~-protein or amyloid A4.
Throughout this document this protein is referred to as ~-amyloid peptide or protein.
~-amyloid peptide is proteolytically derived from a tr~n~emhrane protein, the amyloid precursor protein.
Different splice forms of the amyloid precursor protein are encoded bv a widely expressed gene. see, e.a., K. Beyreuther and B. Mhller-Hill, AnntlAl Reviews in Bioch~mistrv, 58:287-307 (1989). ~-amyloid peptide consists, in its longest forms, of 42 or 43 amino acid residues. J. Kang, et ~1., N~tllre (To~on), 32S:733-736 (1987). These peptides, however, vary as to their amino-termini. C. Hilbich, et ~l., Jol~rn~l of Moleclllar B;oloav, 218:149-163 (1991).
Because senile plaques are invariably surrounded ~y dystrophic neurites, it was proposed early that ~-amyloid peptide is involved in the loss of neuronal cells that occurs in ~l~heim~r's disease. B. Yankner and co-workers were the first to demonstrate that synthetic ~-amyloid peptide could be neurotoxic i n v; tro and in vivo. B.A. Yankner, et ~1., Science, 245:417 (1989); ~ee ~lso, N.W. Kowall, et ~l., Procee~inas of the Nat;on~l Ac~emv of Sciences~ U.S.A., ,

2 1 76 1 ~7 PCT~S9~1146S5 88:7247 (1991). Other research ~roups, however, were unable to consistently demonstrate direct toxicity with ~-amyloid peptide. see. e.a., Nellrohioloav of Aain~, 13:535 (K. Kosik and P. Coleman, eds. 1992). Even groups receiving ~-amyloid peptide from a common source demonstrate conflicting results.
P. May, et al., Nellrohioloav of Aaina, 13:605-607 (1992).
Resolution of this controversy was achieved once a better underst~n~;ng of the role of secondary conformation of ~-amyloid peptide as it relates to neurotoxicity was achieved.
10 Now, numerous researchers repor~ direct neurotoxicity of ~-amyloid peptide.
In addition to Alzheimer's disease, Down~s syndromeis also characterized by an accumulation of ~-amyloid peptide. In patients suffering from Down~s syndrome the ~-amyloid peptide is the primary constituent of senile plaquesand cerebrovascular deposits.
Recently, it has been shown that ~-amyloid peptide is released from neuronal cells grown in culture and is present in cerebrospinal fluid (CSF) of both normal individuals and those persons suffering from Al7he;mer's Disease. See, D. Seubert, et ~l., N~tllre (Ton~on), 359:325-327 (1992).
A possible correlation to the plaque pathology hasbeen developed by several groups demonstrating the direct ~-amyloid peptide-induced neurotoxicity toward cultured neurons.
More recently, in addition to the direct neurotoxicity, an inflammatory response in the ~l7he; m~r ' S
Disease brain, perhaps elicited by ~-amyloid peptide, also contributes to the pathology of the disease. A limited clinical trial with the non-steroidal anti-inflammatory drug indomethacin exhibited a retardation in the progression of Alzheimer's dementia. Rogers et ~l., Sc;~nce, 260:1719-1720 (1993).
Direct neurotoxicity of ~-amyloid peptide was recently reported to be attenuated by co-treatment with beta transforming growth factor (TGF-~). Chao et ~1 , societv of WO95/17095 ~ ~ 2 ~ 7 6 1 2 7 PCT~Sg4114655 .

Nellrosci ences Ahstr~c~s, 19: 1251 (1993). The beta transforming growth factors are multifunctional cytokines produced by many types of cells, including hematopoietic, neural, heart, fibroblast, and tumor cells, that can regulate the growth and differentiation of cells from a variety of tissue origins. Sporn, et ~l., SciPnce, 233:532 (1986).
There are at least five isoforms of TGF-~ currently identified.
TGF-~ has been shown to have numerous reaulatory actions on a wide variety of both normal and neoplastic cells. This protein is multifunctional, as it can either stimulate or inhibit cell proliferation, differentiation, and othr critical processes in cell function. For a general review of TGF-~ and its actions, see Sporn, et ~1., Jollrnal of Cell Bioloav, 105:1039-1045 (1987); Sporn and Roberts, N~ture (rondon), 332:217-219 (1988); and Roberts, et ~l., Recent Proaress in Hormone Rese~rch, 44:157-197 (1988).
In addition to Alzheimer's Disease and other conditions associated with the amyloidogenic peptide ~-amyloid peptide, there exist conditions associated with other amyloidogenic peptides which are structurally si m; 1 Ar to ~-amyloid peptide but which share no sequence homology with the ~-amyloid peptide. Recent studies have demonstrated the functional interchangeability of many of these amyloidogenic peptides with regard to neurotoxicity. P.C.
May, et al., Jol7rnal of Neuroch~mistrv, 61:2330-2333 (1993);
co-p~n~;ng U.S. Patent Application 08/109,782, filed August 19, 1993.
Human amylin (also known as diabetes-associated peptide and islet amyloid polypeptide) is a 37 amino acid peptide first isolated from amyloid deposits in the pancreas of non-insulin-dependent diabetic individuals. C.J.S. Cooper et ~l , Procee~;nas of the N~t;o~l Ac~emv of Sc;encs (USA), 84:8628-8632 (1987). The rat homologue of amylin is 95~
similar in primary sequence to human amylin but has amino acid substitutions in the region con~erring ~-pleated WO95/17095 2 ~ 76 ~ 27 PCT/US94/1465S

secondary structure which render it non-amyloidogenic. See e.a., J. Asai et ~l., Biochemical ~nd ~iophvsic~l Rese~rch commllnic~tions, 164:400-405 (1989)i L.R. Mchean and A.
Balasubr~m~ni~m~ Riochimic~ ~n~ Bio~hvs;ca Act~, 112:317-320 (1992).
~ 2-microglobulin is another protein unrelated to ~-amyloid peptide but capable of forming amyloid deposits in the kidney. [for recent review, ~, J.M. Campistol and M.
Skinner, S~mi~rs in Di~lvsis, 6:117-126 (1993)]. This nonpolymorphic peptide is homologous to the C3 ~om~in of the immlln~globulin class IgG and is one subunit of class I major histocompatibility antigens.
Because of the debilitating effects of Alzheimer's disease, Down's syndrome, and these other conditions associated with amyloidogenic peptides and proteins there continues to exist a need for effective treatments. This invention provides compounds efficacious in the treatment and prevention of these disorders.
This invention ~ncomrasses a method for the inhibition of a physiological disorder associated with an amyloidogenic peptide, which method comprises administering to a mammal in need thereof an effective amount of a compound of Formula I

~5 R

wherein:
A is -O-, -S(O) m~, -N(Rl1)-, -CH2CH2-, or -CH=CH-;

WO95/1709S; 2 1 7 6 ~ 2 7 PCT~ss4/14655 m is 0, 1, or 2;

X is a bond or C1-C4 alkylidenyl;

R~ is a group of the f rmula wherein Rg and R5 are independently C1-C6 alkyl or combine to form, along with the nitrogen to which they are attached, a heterocyclic ring selected from the group consisting o.f hexamethylPnei m; nyl, piperazino, heptamethyl~n~- mi nyl, 4-methylpiperidinyl, imidazolinyl, piperidinyl, pyrrolidinyl, or morpholinyl;

R is hydroxy, halo, hydrogen, C3-C8 cycloalkyl, C2-C7 alkanoyloxy, Cl-C6 alkoxy, or phenyl, said phenyl being optionally substituted with one, two, or three moieties selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, nitro, chloro, fluoro, trifluoromethyl -OSO2-(C1-C1o alkyl) o 3 -OCN-R
or H

R1 is hydroxy, halo, hydrogen, C3 -C8 cycloalkyl, C2-C7 alkanoyloxy, C1-C6 alkoxy, or phenyl, said phenyl being optionally substituted with one, two, or three moieties selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, nitro, WO 95/17095 ~ ' ; 2 ~ 7 6 1 2 7 PCT/US9~/14655 chloro, fluoro, t_ifluoromethyl -OSO2-(C1-Clo alkyl) -oCN-R3 or H

each R3 is independently C1-C6 alkyl, C3-C8 cycloalkyl, unsubstituted or substituted phenyl where the substituent is halo, C1-C6 alkyl or C1-C6 alkoxy;

with the proviso that when X is a bond and A is -S-, R and R1 are not both selected from the group consisting of hydroxy, methoxy, and C2-C7 alkanoyloxy;

or a pharmaceutically acceptable salt or solvate thereof.5 The present invention also provides a method of inhibiting amyloidogenic peptide production comprising A~m;ni~tering to a m~mm~1 in need thereof an effective amount of a compound of Formula I or a pharmaceutically acceptable0 salt thereof.

The present invention also provides a method of increasing TGF-~ expression in the brain, comprising A~m;n; stering to a human in need thereof an effective amount of a compound of Formula I or a pharmaceutically acceptable salt or prodrug thereof.

The present invention also provides a method of inhibiting the inflammatory response associated with Alzheimer's Disease comprising ~m; n; stering to a human in need thereof an effective amount of a compound of Formula I
or a pharmaceutically acceptable salt or prodrug thereof.
The term ~amyloidogenic peptideU refers to those peptides which have the ability to self-associate into higher ordered aggregates and eventually assemble into an amyloid WO95/17095 ~ 2 1 7 6 1 2 7 PCT~S94114655 ~ ! ',,, ', . .

plaque. An especially preferred amyloidogenic peptide target in methods of this invention is the ~-amyloid peptide.
As used herein the term ~inhibit~ or ~inhibiting"
includes its generally accepted me~ning which ~ncompasses prohibiting, preventing, restraining, and slowing, stopping, or reversing progression, severity, or a resultant symptom.
As such, the methods of this invention enc~p~s both therapeutic and prophylactic administration.
The term. ~physiological disorder associated with an amyloidogenic peptide" includes diseases related to the ina~u~riate or undesirable deposition, such as in the brain, liver, kidney, or other organ, of at least one amyloidogenic peptide, and as such includes Alzheimer~s Disease (including familial Alzheimer~s Disease), Down~s Syndrome, advanced aging of the brain, hereditary cerebral hemorrhage with amyloidosis of the Dutch-type tHCHWA-D), and the like.
The term "effective amount~ as used herein refers to the amount of compound necessary to inhibit physiological effects or disorders associated with an amyloidogenic peptide, or inhibit amyloidogenic production or deposition, or inhibit Alzheimer's Disease, as the case may be.
The term "conservative variant~ as used herein refers to amino acid substitutions which alter the primary se~uence of the peptide but do not alter its seCon~ry structure or ~im;nish its amyloidogenic properties.
The terms "human amylin(20-29)u, ~-amyloid11-43)"
and the like refer to amino acids 20-29 of the human amy~in peptide as defined in SEQ ID NO:3 or amino acids 1-43 of ~he ~-amyloid peptide as defined in SEQ ID NO:l. In all such references, the first number listed is the amino-terminus of this peptide sequence.
~-amyloid peptide naturally occurs as a series of - peptides which are 39 to 43 amino acids long, with the shorter, more soluble forms being present in cerebrovascular deposits and the longer forms being found primarily in sènile plaques. F. Prelli, et ~1., Jollrnal of Ne1~roch~m;strv, , WO95/17095 2 1 76 t 27 PCT/US94/14655 51:648-651 (1988). The primary structure of the 43 amino acid long peptide (~1-43) is depicted in SEQ ID NO:l:

Asp Al~ Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln 15 5 Lys Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Gly Ala 30 Ile Ile Gly Leu Met Val Gly Gly Val Val Ile Ala Thr 43 Even though the full length peptide of SEQ ID NO:l: has sufficient solubility in water for the following experiments, for the purposes of convenience, a more water-soluble form of the peptide is often desired. For that reason, the following examples were performed using peptides cont~;ning just the first 40 amino acids of the ~-amyloid peptide (~1-40). The sequence of this preferred peptide is SEQ ID NO:2:
Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln 15 Lys Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Gly Ala 3 0 Ile Ile Gly Leu Met Val Gly Gly Val Val 40 It is understood by those in the art that other fragments of ~-amyloid peptide, comprising amino-truncated, carboxy-truncated, or internal deletions, or any combination of these, may be employed in this invention so long as that peptide fragment demonstrates the requisite neurotoxicity.
Amylin is a 37 amino acid peptide having the primary sequence Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe 15 Leu Val His Ser Ser Asn Asn Phe Gly Ala Ile Leu Ser Ser Thr 3 0

3 0 Asn Val Gly Ser Asn Thr Tyr 37 hereinafter referred to as SEQ ID NO:3.
The peptide ~2-microglobulin has a molecular weight of about 11,000 Daltons. It can be readily purified from urine samples using known techniques. See, e.a., I. Berg-aardr et al., Jollrn~l of B;oloaic~l Chemi strv, 243:4095 (1968).

"C1-C6 alkoxy" represents a straight or branched alkyl chain having from one to six carbon atoms attached to an oxygen atom. Typical C1-C6 alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tibutoxy, pentoxy and the like. The term "C1-C6 alkoxy" includes within its definition the term "C1-C4 alkoxy".
As used herein, the term "C1-C6 alkyl" refers to straight or branched, monovalent, saturated aliphatic chains of 1 to 6 carbon atoms and includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, and hexyl. The term "C10-C6 alkyl" includes The current invention concerns the discovery that a select group of substituted benzofurans, benzothiophenes, Formula I, are useful as in treating or preventing physiological disorders associated with an amyloidogenic peptide.
The terms and abreviations used in the instant examples have their normal "°C" refers to degrees Celsius; "N"
refers to normal or normality; "mmol" refers to millimole or millimoles; "g" refers to gram or grams; "ml" means milliliter or milliliters; "M" refers to molar or molarity;
"MS" refers to mass spectrometry; "IR" refers to infrared spectroscopy; and "NMR" refers to nuclear magnetic resonance spectroscopy.
As used herein, the term "C1C10 alkyl" refers to straight or branched, monovalent, saturated aliphatic chains of 1 to 10 carbon atoms and includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, and hexyl. The term :C1-C10 alkyl"
includes within its definition the terms "C1-C4 alkyl" and "C1-C6 alkyl".
"C1-C6 alkoxy" represents a straight or branched alkyl chain having from one to six carbon atoms attached to an oxygen atom. Typical C1C6 alkoxy groups include methoxy, WO95/17095 i ~ 2 1 76 ~ 27 PCT/US94114655 ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, pentoxy and the like. The term "Cl-C6 alkoxy~ includes within its definition the term "Cl-C4 alkoxy~.
~Cl-C6 alhylidenyl~ refers to a straight or branched, divalent, saturated aliphatic ch~in~ of 1 to 6 carbon atoms and includes, but is not limited to, methylenyl, ethylenyl, propylenyl, isopropylenyl, butylenyl, isobutylenyl, t-butylenyl, pen~ylenyl, isopentylenyl, hexylenyl, and the like. The term "Cl-C4 alkylidenyl~ is encompassed within the term ~Cl-C6 alkylidenyl~.
The term ~halo~ encompasses chloro, fluoro, bromo and iodo.
The term "leaving group~ as used herein refers to a group of atoms that is displaced from a carbon atom by the attack of a nucleophile in a nucleophilic substitution reaction. The term "leavin~ group~ as used in this document encompasses, but is not limited to, activating groups.
The term "activating group'~ as used herein refers a leaving group which, when taken with the carbonyl (-C=O) group to which it is attached, is more likely to take part in an acylation reaction than would be the case if the group were not present, as in the free acid. Such activating groups are well-known to those skilled in the art and may be, for example, succinimidoxy, phth~lim;doxy, benzotriazolyloxy, benzenesulfonyloxy, methanesulfonyloxy, toluenesulfonyloxy, azido, or -O-CO-(C4-C7 alkyl).
Many of the compounds employed in the present invention are derivatives of naphthalene which are named and numbered according to the RING INDs, The American Chemical Society, as follows.

WO95/17095 r - ~ r I 2' F 7 6 t 27 PCT/US9~/14655 ~ 11 ~

In a similar manner some of the compounds employed in the present invention are derivatives of 1,2-dihydronaphthalene which are named and numbered according to the RING INDEX as follows.

q ~

Many of the compounds of the present invention are derivatives of benzofuran which are named and numbered according to the RING INDEX, The American Chemical Society, as follows.

~0 Some of the compounds of the present invention are derivatives of benzo[b]thiophene which are named and numbered according to the RING INDEX as follows.
X~
In a similar m~nner some of the compounds of the present invention are derivatives of indole which are named and numbered according to the RING INDEX as follows.

H

WO 95/17095 1 : i 2 1 7 6 1 2 7 PCT/US94/14655 The more preferred com~ounds employed in the methods of this invention are those compounds of Formula I
wherein a) A is -O-, -S-, -CH2-CH2-, or -CH=CH-i b) R is hydrogen, hydroxy, C1-C3 alkoxy, or -OSO2-(Cl-C10 alkyl);
c) R1 is hydrogen, hydroxy, C1-C3 alkoxy, or -OSO2-(C1-C1o alkyl);
d) X is a bond or methylene; and e) R2 is piperidinvl, hexamethyleneiminyl, pyrrolidinyl, or -NR4R5, where R~ and R5 are C1-C4 alkyl; and the pharmaceutically acceptable acid addition salts and solvates.
The most preferred compounds employed in the methods of this invention are those compounds of Formula I
wherein a) A is -S-;
b) R is hydrogen, hydroxy, C1-C3 alkoxy, or -oSO2-(C1-C1o alkyl);
c) Rl is hydrogen, hydroxy, Cl-C3 alkoxy, or -oSO2-(C1-C1o alkyl);
d) X is a bond or methylene; and e) R2 is piperidinyl, hexamethyleneiminyl, pyrrolidinyl, or -NR4R5, where R4 and R5 are C1-C4 alkyl; and f) at least one of R and R1 is -OSO2-(C1-C1o alkyl);
and the pharmaceutically acceptable acid addition salts and solvates thereof.
The compounds of the present invention can be prepared by a variety of procedures well known to those of ordinary skill in the art. The particular order of steps re~uired to produce the compounds of Formula I is dependent upon the particular compound being synthesized, the starting compound, and the relative lability of the substituted moieties.
A. Preparation of Dihydronapthalenyl Compounds ~ = ~

WO 95/17095 ~ ~ . ` ~ 2 ~ 7 6 1 Z 7 PCT/lJS94/1465~;

The compounds employed in the present invention in which A is -CH2-CH2- or -CH=CH- may be prepared essentially as descri~ed in U.S. Patent 4,230,862, issued to T. Suarez and C.D. Jones on October 28, 1990, which is herein incorporated by reference.
These compounds are generally prepared by the following sequences, the dihydronaphthalene structures in general being precursors to the napththalene compounds.
The naphthalenes and dihydronaphthalenes employed `n the methods of the instant invention may be prepared ~y reacting a tetralone of Formula II

R~

II
in which Rc is hydrogen, Cl-C6 alkoxy, or benzyloxy with a phenyl benzoate of Formula III

Il ~3~o-~

III

in which yl is methoxy, benzyloxy, or ~O-(CH2)n-NR~Rb, where n is 1-6, and -NR~Rb is R2. This reaction is generally carried out in the presence of a moderately strong base such as sodium amide and at room temperature or ~elow.
The product which is obtained is a substituted tetralone of Formula IV.

WO95/17095 ; - ;. 2 1 7 6 1 2 7 PCT~S94/14655 yl C=O
RC ~

IV

This substituted tetralone is then reacted under Grignard reaction conditions with the Grignard reagent of the formula Rla ~ Y~ MgBr in which Rla is hydrogen, Cl-C6 alkoxy, or benzyloxy and ya is a bond, methylene, or ethylene.
The compounds which are produced,a 3-phenyl-4-aroyl-l,2-dihydronaphthalenes, have the following formula, Formula V.
~yl C=O

RC ~ ~ Rla In those instances in which yl is methoxy, a compound of Formula V can be treated with pyridine hydrochloride at reflux to produce the corresponding hydroxy compound. Under these conditions, should Rc or Rla be alkoxy WO95/17095 ' ` 2 1 76 ~ 27 PCTIUS94/14655 or benzyloxy, these groups will also be cleaved, resulting in hydroxy groups, In those instances in which yl is methoxy or benzyloxy, and Rc or Rl~ is alkoxy or benzyloxy, the group at yl can be selectively cleaved by treating a compound of Formula V with an equivalent of sodium thioethoxide in N,N-dimethylformamide at a moderately elevated temperature of about 80C to about gOC. The process of the selective cleavage may be monitored by periodic thin layer chromatography analysis. The reaction is complete when little or no starting material rem~i n~, Once the compound of Formula V in which yl has been converted to hydroxy has been generated, that compounds can then be treated with a compound of Formula VII
L-(CH2)n-NR~R~
VII

wherein L is a good leaving group such as halo, especially chloro. Under the usual reaction conditions, of course, alkylation will be effected at each of the unprotected hydroxy groups which are present in the molecule. This can be avoided, and alkylation at the 4-ben20yl groups alone can be achieved, by carrying out the reaction in the presence of an excess of finely powdered potassium carbonate and using an equivalent or slight excess of the compound of Formula VII.
Dep~i n~ upon the intended structure of the final product, the compound contAin;ng the substituent of Formula VII can then be further treated with an additional quantity of sodium thioethoxide in N,N-dimethylformamide as aforedescribed to effect cleavage of any rP~Aining alkoxy or benzyloxy groups, thereby providing another sequence for achieving formation of those compounds employed in this invention in which Rl and/or R2 are hydroxy.
In any of the above, it is evident that the particular se~uence of synthetic steps designed to produce a compound having substituents of particular definition and WO95/17095 ; ;-- 21 76 1 ~7 PCT/US94/14655 location is such as one of ordinary skill in the art will well recognize.
In another route for preparing the compounds of Formula I, compounds of Formula VI

RaRbN- ( CH2 ) n~~

R2a /~yc VI

wherein: R2a is -H or C1-C6 alkoxy; and yc is Cl-C6 alkoxy-substituted phenyl or benzyl, are prepared essentially as described by C.D. Jones, et ~1., Jollrn~l of Me~;c; n~l ~hPm;strv, 53:931-938 (1992), which is herein incorporated by reference.
Generally, a tetralone, as described above, or a salt thereof, is acylated using st~n~rd Friedel Crafts conditions to provide a highly enolized diketone of formula VIa R2a ~ ~ O-C ~ OCH3 H3C ~ H3C ~

R2a ~ R2a ~ OH

VIa PCT~Sg4/14655 WOg5/17095 ~.~ ~ ~ ;~..
2 1 76 ~ 27 wherein R2a is -H or Cl-C6 alkoxy.
Subsequent deriva~ization using sodium hydride, followed by the addition of diphenyl chlorophosphate, gives the enol phosphate derivative tentatively assigned the Formula VIb ~0 R2a ~ opo(oPh)2 VIb ~0 wherein ~2a iS as de~ined a~ove.
Addition of phenyl- or benzyl-, substituted phenyl-or substituted benzylmagnesium bromide to a compound of formula VIb, and su~sequent selective demethylation provide compounds of formula VIc and VId, respectively, as described by Jones, sl~nr~.

R2a J~ R2aJ~yc VIc VId wherein R2A and yc are as defined a~ove.
Finally a compound of formula VId is alkylated with a compound of the formula L-(CH2)n-NR~Rb , WOg5/17095 2 1 7 6 1 2 7 PCT~Sg4/14655 ln which L is a bromo or, preferably, a chloro moiety, and R2a and yc optionally are dealkylated by standard procedures, to provide compounds of formulae VIe and VIf, respectively.

R~R~N- ( C~z ) n~~ RaRbN- ( CH ~ ) n~~

R2a J~ R2b J~
VIe VIf wherein R2b is -H or -OH and yd is phenyl, benzyl, hydroxyphenyl, or hydroxybenzyl.
In the process for preparing compounds of formula VIe or VIf, it is evident that the particular sequence of synthetic steps designed to produce a compound having substituents of par~icular definition and location is such as one of ordinary skill in the art will recognize.
The compounds of Formula VIf can be substituted using stAn~rd means, if desired, to produce the corresponding dihydronaphthenyl compounds of Formula I.

B. Preparation of Napthalenyl Compounds Those compounds of Formula I which are substituted naphthalenes are readily prepared from the correspo~in~
dihydronaphthalenyl compounds. Selective dehydrogenation of the dihydronaphthalene structure to produce specifically the correspo~;n~ naphthalene can be accomplished by treatment with 2,3-dichloro-5,6-dicyano-l,4-benzo~lino~e ~DDQ) at a temperature of from about 50C to about 100C. The naphthalene which is produce may be further col~ve Led to other naphthalene compounds by means of the derivatizing reactions described ~l~r~.

=
WO 95/17095 ~. ~ 2 1 7 6 ~ 2 7 PCT/US9~/14655 ~,nl e Preparation of 3-(4-methoxyphenyll-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl-1,2-dihydronaphthalene, citrate salt The title compound was prepared as described in United States Patent 4,230,862. To a suspension of sodium amide (15.2 g, 0.38 mol) in 2~0 ml o~ tertrahydrofuran were added 50 grams (0.34 mol) of ~-tetralone. The mixture was stirred for 15-20 minutes, and 78 grams of phenyl p-methoxybenzoate dissolved in tetrahydrofuran were added. The temperature of the reaction mixture was maintained below 10C, and the mixture was then stirred at room temperature overnight. The reaction mixture was concentrated and the water was added to the residue. The aqueous mixture was extracted with ethyl acetate, and the ethyl acetate extract was washed and concentrated.
The residue was chromatographed on silica using benzene as eluant. The purer fractions obtained by the chromatographic separation were combined and concentrated, and the residue was dissolved in a ~in;m~m of methanol. The methanol was cooled, and 35.2 grams of 1-(4-methoxybenzoyl)-2-tetralone were collected by ~iltration.

4-Bromoanisole (18.7 g, 0.1 mol) was added dropwise in ether to tetrahydrofuran cont~;~ 7 ng 5 drops of 1,2-dibromoethane and 3.6 grams (0.15 mol) o~ magnesium.
Reaction occurred almost ;mme~;~tely, and the addition was continued at a slow rate with evolution of heat sufficient to maintain a general re~lux. Upon completion of the addition, the above substituted ~-tetralone dissolved in acetone was added dropwise with stirring over a two hour period, the mixture ~eing maintained at 40C. The resulting mixture was then poured into cold, dilute hydrochloric acid, and the acidic mixture was extracted with ethyl acetate. The ethyl acetate extract was washed, dried, and concentrated to an oil. The oil was chromatographed over silica using benzene as eluant. A subsequent elution of the column with a mixture

5 ~ . 2 t 7 6 1 2 7 PCT/US94/14655 , ---- GO

of benzene containing two percen~ ethyl acetate yielded 15 grams of 3-(4-methoxyphenyl)-4-(4-methoxybenzoyl)-1,2-dihydronaphthalene as an oil.
A mixture of 11.1 grams (0.03 mol) of the above dimethoxy product, 7.2 grams of sodium hydride (50 percent in oil), and 11 ml of ethyl mercaptan in N,N-dimethylformamide was prepared. The mixture was heated to 65-70C and maintained at that temperature for about two hours. The mixture was then cooled and cone~rated. The concentrate was acidified and extracted with ethyl acetate. The ethyl acetate extract was washed, dried, and evaporated. The residue was dissolved in benzene and chromatographed over silica to obtain five grams of an oil comprising relatively pure 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene.
The above phenolic product (4.3 g, 0.01 mol) wasdissolved in N,N-dimethylformamide. To this solution was added 0.7 grams of sodium hydride (50 percent in oil), and the resulting mixture was warmed to 40C for one hour and then was cooled to room temperature. To the mixture then were added 1.6 grams of 1-chloro-2-pyrrolidinylethane, and the mixture was warmed to 60C and maintained at this temperature for about two hours. The reaction mixture was then stirred at room temperature overnight.
The mixture was concentrated, and water was added to the residue. The aqueous mixture was extracted with ethyl acetate. The ethyl acetate extract was washed and concentrated to a residue. The residue was extracted with hexanes, the insoluble portion was dissolved in ethyl acetate, and the etllyl acetate solution was extracted with 1 N hydrochloric acid. The acid extract was rendered alkaline, and then was extracted with ethyl acetate. The ethyl acetate extract was washed and concentrated. One equivalent of citric acid in acetone then was added to the concentrate, and the mixture was concentrated to dryness. The residue was dissolved in a large volume of methyl ethyl ketone. The ketone solution was concentrated to a~out 300 ml and was wo 95/17095 ~ 6 t 2 7 PcT/uss4ll46s5 cooled to 0C. The title product, the citrate salt of 3-(4-methoxyphenyl)-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl-1,2-dihydronaphthalene, was collected by filtration and vacuum dried. mp 82-85C.
5 ~nalysis for C36H39NlO:
Theory: C, 66.96; H, 6.09; N, 2.17; O, 24.78.
Found: C, 66.70; H. 6.27; N, 2.27; O, 24.54.

~ mn 1 e 2 Preparation of 3-phenyl-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl~-7-methoxy-1,2-dihydronaphthalene.

The title product was prepared as described iin United States Patent 4,230,862. To 300 ml of N,N-dimethylformamide were added 107 grams of phenyl p-hydroxybenzoate and 26 grams of sodium hydride (50 percent in oil). The mixture was heated to 60C and maintained at this temperature for about two hours. To this mixture was added 1-chloro-2-pyrrolidin-1-ylethane (67 g), and the mixture was stirred overnight at 85C. The bulk of the N,N-dimethylformamide then was evaporated from the mixture.
Water was added to the residue, and the aqueous mixture was extracted with ethyl acetate. The ethyl acetate extract was concentrated, and the residue was dissolved in a 1:1 mixture of ether and ethyl acetate. The organic solution was then extracted with 2 N hydrochloric acid, and the acid extract was added dropwise to 2 N sodium hydroxide. The resulting mixture was extracted with ethyl acetate, and the ethyl acetate extract was washed and then dried over r~ nesium sulfate. The ethyl acetate was concentrated to obtain 110 grams of crude phenyl p-(2-pyrrolidin-1-ylethoxy)~enzoate.
To a suspension of 20 grams (0.5 mol) of sodium amide in tetrahydrofuran were added dropwise 41.7 grams of 6-methoxy-2-tetralone in tetrahydrofuran, the temperature of the mixture being maintained below 10C. Upon completion of the addition, the mixture was stirred for 20 minutes, the -WO 95/17095 5 ~ 2 ~ 7 6 1 2 7 PCT/US94114655 reaction mixture being maintained below 10C, after which time an exothermic reaction occurred, the reaction temperature rising ~o about 20C.
The above prepared phenyl p-(2-pyrrolidin-1-ylethoxy)benzoate, dissolved in tetrahydrofuran, was thenadded dropwise, and the mixture was stirred overnight at room temperature. The mixture was poured into water, and the resulting mixture was extracted with ethyl acetate. The ethyl acetate extract was washed several times with water, and dried over magnesium sulfate. The ethyl acetate was concentrated to obtain about 100 grams of crude material which was dissolved in 1.5 liters of acetone, and one equivalent of citric acid in 400 ml of ethyl acetate was added. The resulting solid was isolated by filtration and vacuum dried to obtain 85.9 grams of 6-methoxy-1-[4-(2-pyrrolidin-1-ylethoxy)benzoyl]-2-tetralone. The product was then chromatographed over silica using ethyl acetate as eluant, and the citrate salt was prepared from the recovered product.
The above product (8.6 g, 0.02 mol) was added to a solution of phenylmagnesium bromide in tetrahydrofuran. The resulting mixture was stirred for one hour at room temperature and then was warmed to 50C and maintained at this temperature for three hours. The resulting mixture was poured into a mixture of ice and hydrochloric acid, and the acid mixture was extracted with ethyl acetate. The ethyl acetate extract was washed, dried, and concentrated to obtain 10.5 grams of a red-brown oil. The oil was added to 500 ml of acetic acid, and the mixture was heated on a steam bath for about 30 minutes. The acid was stripped off, and water as added to the residue.
The aqueous mixture was rendered alk~line by addition of base, and the alk~l;ne mixture was extracted with ethyl acetate. The extract was dried and concentrated to obtain 8.7 grams of product which was dissolved in acetone, and one equivalent of citric acid was added to the mixture.
The acetone was stripped off, and methyl ethyl ketone was WO 95/17095 ` 2 t 7 6 t 2 7 PCT/US94/14655 . = ~'C . .

added to the residue. The mixture was maintained at 0C
overnight, and the crystals which formed were collected by filtration and washed with cold methyl ethyl ketone and vacuum dried. The solid was recrystallized from acetone to obtain the title compound in the form of its citrate salt.
- mp 98-100C.
AnalysiS of C36H3sNolo:
Theory: C, 6~.96; H, 6.09; N, 2.17; O, 24.78.
Found: C, 66.72; H, 6.27; N, 2.09; O, 24.50.
The title compound in the form of its free base was generated by treatment of the citrate salt with dilute alkali.
Analysis for C30H3lNO5:
Theory: C, 79.44; H, 6.89; N, 3.09.
Found: C, 79.19; H, 6.68; N, 2.91.

~x~mnl e 3 Preparation of 3-phenyl-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl]-1,2-dihydronaphthalene The title product was prepared as described in United States Patent 4,230,862. To a solution of 5.0 grams (18 mmol) of 1-(4-methoxybenzoyl)-2-tetralone (prepared as described in Example 1) in 50 ml of ether was added dropwise at 0C a solution of phenylmagnesium bromide (18 mmol) in 9 ml of ether. Upon completion of the addition, the mixture was stirred for twenty minutes. Thin layer chromatography of the reaction mixture indicated the presence of starting material. An additional 13.5 ml o~ the phenylmagnesium bromide solution were added.
The mixture was refluxed for two hours and then was cooled and poured over iced aqueous ~mm~nium chloride s~lution. The organic layer was separated and washed with brine. The mixture was then dried over m~gnesium sulfate, filtered, and evaporated to give about ten grams of a yellow :
WO 95/17095 . . . ~ PCT/US94/14655 2~76127 oil. After a wash with hexanes, the product was further purified by chromatography ~o give 4.67 grams of 3-phenyl-4-(4-methoxybenzoyl)-1,2-dihydronaphthalene.
To 2.0 grams (6 mmol) of the above dihydronaphthalene, dissolved in 10 ml of N,N-dimethylformamide, were added sodium thioethoxide (7.5 mmol), dissolved in 15 ml of N,N-dimethylformamide. The addition was carried out under a nitrogen atmosphere and at 80C. The mixture was maintained at 80C for fifteen hours. The mixture was then cooled and poured in~o an iced aqueous ammonium chloride solution. The resulting mixture was extracted with ethyl acetate, and the ethyl acetate extract was washed four times with brine.
The ethyl acetate extract was dried over magnesium sulfate an evaporated to give an oil which was further purified by chromatography on a silica column, using benzene to elute impurities. The product was then eluted with ethyl acetate to give, upon evaporation of the ethyl acetate, 1.69 grams of 3-phenyl-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene as a clear pale yellow oil.
A mixture of 1.61 grams (4.95 mmol) of the above product in 10 ml of dry N,N-dimethylformamide cont~;n;ng 119 mg (4.95 mmol) of sodium hydride and freshly distilled 1-chloro-(2-pyrrolidin-1-yl)ethane. The addition was made under a nitrogen atmosphere with the temperature being maintained at about 10C. Upon completion of the resulting efferverscence, the mixture was heated to 80C and maintained at that temperature for about two hours. The mixture was then poured into water, and the total was extract with ether.
The ether extract was washed five times with brine, and dried over magnesium sulfate. The ether layer was then filtered and evaporated to give a gray oil, which was further purified by chromatography to give 3-phenyl-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl]-1,2-dihydronaphthalene.
The product was converted to the corresponding citrate salt by treatment with 0.59 grams of citric acid in 50 ml of hot acetone. The resulting mixture was evaporated WO 95/17095 ' " i ~ ~ 7 6 T 2 7 PCT/US94/146~!i ~o dryness, and the residue was stirred for about fifteen hours with ether to obtaln the cltrate salt. mp 89-93C.
Analysis for C33H37N09 0.5 H2O:
- Theory: C, 67.34: H, 6.13; N, 2.25.
Found: C, 67.06; H, 6.41; N, 2.66.

~.x~mnl e 4 Preparation of 1-~4-(2-pyrrolidin-1-ylethoxy)benzoyl]-2-phenylnaphthalene, citrate salt The title product was prepared as described in United States Patent 4,230,862. To 30 ml of dioxane were added 3-phenyl-4-(4-methoxybenzoyl)-1,2-dihydronaphthalene (1.90 g, 5.58 mmol), prepared as described in Example 3, sllnr~, and 2~3-dichloro-5~6-dicyano-1~4-benzo~;non~ (2.00 g, 8.81 mmol). The resulting mixture was heated to reflux and refluxed for twelve hours under a nitrogen atmosphere. The mixture was then cooled and evaporated to dryness. The residue was partitioned between ether and water. The organic fraction was washed 5 N sodium hydroxide (5 x 20 ml), followed by a wash with brine. The mixture was then dried over magnesium sulfate and evaporated to give 1.9 grams of substantially pure 1-(4-methoxybenzoyl)-2-phenylnaphthalene.
Employing substantially the same demethylation procedure as described in Example 3, 1.83 grams (5.41 mmol) of the above product were treated with sodium thioethoxide to obtain 1.4 grams of 1-(4-hydroxybenzoyl)-2-phenylnaphthalene.
To 10 ml of N,N-dimethylformamide were added 1.25 grams of the above product. The resulting mixture was added at about 10C to a mixture of 20 ml of N,N-dimethylformamide cont~;n;n~ 120 mg (5.0 mmol) of sodium hydride and 800 mg of 1-chloro-2-(pyrrolidin-1-yl~ethane. Upon completion of the resulting effervescence, the mixture was heated to 80C and maint~;ne~ at that temperature for about three hours, during which time sodium chloride precipitated. The mixture was cooled and evaporated to dryness. The resulting residue was WO95/17095 ~ 2 1 76 1 ~7 PCT~Sg~114655 ~

partitioned between water and ethyl acetate. The organic fraction was washed with brine (5 x 25 ml). The organic fraction was dried and evaporated to give 1.62 grams of 1-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-2-phenylnaphthalene as a yellow oil.
The above free base was converted to the corresponding citrate salt in accordance with the method of Example 3, employing 0.811 grams of citric acid hydrate. The title compound was obtained as an amorphous solid which crystallized on standing overnlght in ether. mp 105-108C.
Analysis for C33H3sMOg H2O:
Theory: C, 65.55; H, 5.90; N, 2.22.
Found: C, 66.90; H, 5.85; N, 2.25.

F~mnle 5 Preparation of 3-(4-methoxyphenyl)-4-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, citrate salt.

The title compound was prepared as described in United States Patent 4,230,862. To a suspension of sodium hydride (0.269 g, 11 mmol), washed free of mineral oil, and 1-chloro-2-(piperidin-1-yl)ethane (1.82 g, 12 mmol) in N,N-dimethylformamide (50 ml) at 0C, and under a nitrogen atmosphere, were added 4.0 grams (10 mmol) of 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene, prepared as described in Example 1, dissolved in 20 ml of N,N-dimethylformamide. The solution was added dropwise with stirring. When the effervescence had ceased for the most part, the mixture was heated to 50C and m~;nt~ine~ at that temperature for several hours. The progress of the reaction was monitored by thin layer chromatography.
Once the reaction had progressed suf~iciently, the N,N-dimethylformamide was evaporated, and the concentrated mixture was poured over ice water and ethyl acetate. The ethyl acetate fraction was washed with brine, dried over potassium carbonate, ~iltered, and evaporated, The resulting - ~ t 7 6 ~ 2 7 PCTIUS94/14655 oil was chromatographed over a 1.5" x 12" silica column using the following as a double gradient:

(i) 10 percent ethyl acetate in ~enzene (500 ml) 20 percent ethyl acetate in benzene (2 liters);
(ii) 20 percent ethyl acetate in benzene (1.5 liters) ~ 1:1 mixture of methanol and ethyl acetate (1.5 liters).

The appropriate frac~ions were concentrated to give an almost colorless oil. The oil was dissolved in ethyl acetate, and the ethyl acetate solution was dried over potassium carbonate, filtered, and evaporated to give 4.7 grams of the free base of the title compound as a pale yellow5 oil.
The free ~ase (3.4 g, 7.28 mmol) was treated with citric acid monohydrate (1.49 g, 7.1 mmol) in about 20 ml of boiling acetone. When a clear solution was obt~ine~, the acetone was evaporated, 300 ml of anhydrous ether was added, and the resulting precipitate was stirred overnight. The title compound (5.2 grams) was collected as a white powder.
AnalysiS for C37H41NO1o:
Theory: C, 67.36; H, 6.26; N, 2.12.
Found: C, 67.25; H, 5.96; N, 1.84.
~x;~mn 1e 6 Preparation of 3-(4-methoxyphenyl)-4-[4-(2-dimethyl ~m noethoxy)benzoyl~-1,2-dihydronaphthalene, citrate salt.

The title compound was prepared as described in United States Patent 4,230,862. To 50 ml of acetone were added 4.0 grams (11.2 mmol) of 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene, prepared as described in Example 1, 1.81 grams (16.8 mmol) of 1-chloro-2-dimethylaminoethane (freshly prepared from the hydrohloride), WO95117095 ` - 2 1 76 1 27 PCT~Sg~/146~5 ~

and 2.32 grams (16.8 mol) of finely powdered potassium chloride. The resulting mixture was refluxed under nitrogen with stirring for about 72 hours. The progress of the reaction was monitored by thin layer chromatography.
The resulting mixture was then poured over ice, and the resulting mixture was extracted with ether. The ether was washed three times with brine, dried over potassium carbonate, filtered, and evaporated to obtain 4.51 grams of the free base of the title compound as a brown oil.
The oil was vacuum dried and then was converted to the citrate salt by treatment with 2.17 grams (10.4 mmol) of citric acid monohydrate in 50 ml of hot acetone. Evaporation of the acetone and stirring of the residue with ether gave 5.2 grams of the title compound as an amorphous solid.
Analysis for C34H37NOlo:
Theory: C, 65.90; H, 6.02; N, 2.26.
Found: C, 66.17; H, 6.23; N, 2.37.

~x~mnl e 7 Preparation of 3-(4 hydroxyphenyl)-4-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, mesylate salt The title compound was prepared as described in United States Patent 4,230,862. To 25 ml of methyl ethyl ketone were 10 grams (2.92 mmol) of 3-(4-hydroxyphenyl)-4-(4-hydroxybenzoyl)-1,2~dihydronaphthalene, 0.497 grams (2.92 mmol) of l-chloro-2-(pyrrolidin-1-yl)ethane, and 1.21 grams (8.77 mmol) offinely powdered potassium carbonate. The resulting mixture was refluxed for 16 hours. The mixture was then cooled and poured into a mixture of water and ethyl acetate. The resulting mixture was rendered acidic by addition of 1 N hydrochloric acid and then alk~l;ne by the addition of sodium bicarbonate.
The organic fraction was washed with brine, dried over m~nP~ium sulfate, and evaporated to give a yellow oil.
The resulting oil was further purified by chromatography.

woss/~70ss ~ 7 6 1 2 7 PCT~ss4/l4655 The free base (362 mg, 0.825 mmol) as converted to the mesylate aslt by treatment with an equivalent of methanesulfonic acid in acetone to yield the title compound ~ as an amorphous solid.
~nalysis for C31H37NO6S:
Theory: C, 67.27; H, 6.21; N, 2.61.
Found: C, 67.25; H, 6.19; N, 2.69.

~X~ mn 1 e 8 Preparation of 3-(4-methoxyphenyl)-4-~4-~2-(hexamethyleneimin-1-yl)benzoyl]-1,2-dihydronaphthalene, mesylate salt The title compound was prepared as described in United States Patent 4,230,826. To 50 ml of methyl ethyl ketone were added 3.0 g (8.43 mmol) of 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene, 1.84 g (9.27 mmol) of 1-chloro-2-(hexamethyleneimin-1-yl)ethane hydrochloride, and 3.25 grams (25.3 mmol) of finely powdered potassium carbonate. The mixture was refluxed for 48 hours.
The mixture was then poured into water, and ethyl acetate was added. The resulting organic layer was separated, washed with brine, dried, and evaporated to a yellow oil. The oil was ~urther purified by chromatography.
The free base of the title compound was recovered (2.51 g) as a pale yellow oil. The oil was treated with 0.431 g (4.48 mmol) of meth~ne~ulfonic acid in 10 ml of acetone. Upon scratching and cooling of the mixture, crystals formed. The mixture was cooled overnight and 1.97 grams of the title compound were obtained as a white crystals. mp 123-125~C.
Analys iS for C34H4lNO6S:
Theory: C, 68.61; H, 6.80; N, 2.42.
Found: C, 68.38; H, 6.62; N, 2.40.

WO95/17095 ` ~ 76 1 27 PCT/US9~/1465~i ~.~rAmnle 9 Preparation of 3-(4-methoxyphenyl)-4-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, mesylate salt The title compound was prepared as described in United States Patent 4,230,862. To 150 ml of methyl ethyl ketone were added 7.8 g (21.9 mmol) of 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene, 4.84 grams (23.6 mmol) of 1-chloro-2-(piperidin-1-yl)ethane hydrochloride, and 14.S grams (109 mmol) of potassium carbonate. The resulting mixture was refluxed overnight.
The mixture was then poured into a mixture of water and ethyl acetate. The resulting orgnaic fraction was spearated, washed with brine, dried over magnesium sulfate, and evaporated in vacuo to obtain the free base of the title compound as a yellow oil.
The oil was dissolved in 30 ml of acetone and was treated with 2.105 grams (21.9 mmol) of methanesulfonic acid.
The mixture was cooled and scratched, and the title compound was collected at -40C and ashed well with acetone and ether cooled to about -60C. The solid was then vacuum dried at 100C to obtain 11.21 grams of the title compound as a white crystalline solid. mp 157-158C.5 Analysis for C33H3gNO6S:
Theory: C, 68.18; H, 6.62; N, 2.48.
Found: C, 68.11; H, 6.76; N, 2.50.

~x~mn le 10 Preparation of 3-(4-methoxyphenyl)-4-(4-diethyl ~m; n~ethoxybenzoyl)-1,2-dihydronaphthalene, mesylate salt To 75 ml of methyl ethyl ketone were added 4.0 grams (11.2 mmol) of 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene, 2.41 grams (14 mmol) WO 95/17095 r ~ 7 6 1 2 7 PCT/US94/14655 ~ 31 ~

of 1-chloro-2-diethylaminoethane hydrochloride, and 7.93 grams (56 mmol) of finely powdered potassium carbonate. The mixture was refluxed overnight, and, employing the method of Example 9, 5.67 grams of the free base of the title compound were obtained as a yellow oily material.
The oil was treated with 1.07 grams (11.2 mmol) of meth~ne~ulfonic acid in about 15 ml of acetone. The resulting mixture was maintained with cooling for several days after which white crystals appeared. The crystals were somewhat hygroscopic and were collected as quickly as possible and vacuum-dried. There were obtained 4.3 grams of the title compound as a white c~ystalline solid.
AnalysiS for C31H3gNO6S:
Theory: C, 67.24; H, 7.10; N, 2.53.
Found: C, 67.48; H, 6.92; N, 2.43.

~x~mnle 11 Preparation of 3-(4-methoxyphenyl)-4-(4-diiso~o~laminoethoxybenzoyl)-1,2-dihydronaphthalene, mesylate salt To 75 ml of methyl ethyl ketone were added 3.84 grams (10.8 mmol) of 3-(4-methoxyphenyl)-4-(4-hydroxybenzoyl)-1,2-dihydronaphthalene, 2.70 grams (13.5 mmol) of 1-chloro-2-diiso~o~ mi noethane hydrochloride, and 7.11 grams (S4 mmol) of finely powdered potassium carbonate.
The mixture was allowed to reflux overnight, and, upon workup, in accordance with the procedure of Example 9, 5.64 grams of the free base of the title compound were obtained as a yellow oily substance. The oily product was treated with 1.04 grams (10.8 mmol) of methAneculfonic acid in about 25 ml of acetone. The mixture was cooled, and c~stals slowly appeared. The crystals collected at -40C with the aid of acetone cooled to -60C. Vacuum drying of the product gave 5.1 grams.
Analysis for C33H41NO6S:

WO 95117095 r-~ e PCT~Sg~/1465 Theory: C, 68.37; H, 7.31, N, 2.42.
Found: C, 68.08; H, 6.91; N, 2.21.

The following compounds were prepared essentially as described in the above examples:

~x~mnle 12 3-hydroxy-4-[4-t2-(pyrrolidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, sodium salt F.xam~le 13 2-(4-methoxyphenyl)-1-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]naphthalene, mesylate salt Fx~mnle 14 3-(4-methoxyphenyl)-~4-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-7-methoxy-1,2-dihydronaphthalene, mesylate salt Fx~mnle 15 3-(4-methoxyphenyl)-4-t4-(2-dimethyl ~mi noethoxy)benzoyl]-l~2-dihydronaphthalene, 2-hydroxy-1,2,3-propanetricarboxylic acid salt Fx~mnle 16 3-(4-methoxyphenyl)-4-[4-[2-(N-methyl-l-pyrrolidinium)ethoxy]benzoyl]-1,2-dihydronaphthalene, iodide salt Fx~mnle 17 3-(4-methoxyphenyl)-4-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaPhthalene~ mesylate salt WO95/17095 `. , ~. ~ 2 i 7 6 PCT/US94/14655 C. Preparation of Indoles, Benzofurans and Benzothiophenes The benzofurans, benzothiophenes and indoles employed in the methods of the instant invention were made essentially as described in United States Patents 4,133,814, issued January 9, 1979, 4,418,068, issued November 29, 1983, and 4,380,635, issued April 19, 1983, all of which are herein incorpora~ed by reference. This process provides a convenient process which acylates a methylated starting compound and then optionally demethylates it to obtain the desired dihydroxy product. The acylation and demethylation may be performed in successive steps in a single reaction mixture or the intermediate may be isolated and the demethylation step be performed in a separate reaction.
The methyl-protected compound of Formula VII

H3CO ~ OCH3 VII

is most easily obtained by reacting 3-methoxyphenol and a-bromo-4-methoxyacetophenone in the presence of a strong base at a relatively low temperature, to form ~-(3-methoxyphenoxy)-4-methoxyacetophenone, which is then ring closed with an agent such as polyphosphoric acid at a high temperature to obtain the intermediate compound of Formula VII.
The acylation of this invention is a Friedel-Crafts acylation, and is carried out in the usual way, using alllm;~llm chloride or bromide, preferably the chloride, as the acylation catalyst.
The acylation is ordinarily carried out in a solvent, and any inert organic solvent which is not significantly attacked by the conditions may be used. For WO 95/17095 ~ .. 2 ~ 7 6 1 2 7 PCT/US94/14655 i example, halogenated solvents such as dichloromethane, 1,2-dichloroethane, chloroform, and the like may be used, as can aromatics such as benzene, chlorobenzene, and the like. It is preferred to use a halogenated solvent, especially dichloromethane.
It has been found that toluene is rather easily acylated under the conditions used in the Friedel-Crafts acylation, and so it is important, when toluene is used in an earlier step of the process, to remove it as completely as possible from the protected starting compound, to avoid wasting the acylating agent.
The acylations may be carried out at temperatures from about -30 C to about lOO C, preferably at about ambient temperature, in the range of from about l5 C to about 30 C.
The acylating agent is an active form of the appropriate benzoic acid of Formula VIII

:Ra- C~ - ( CH2 ) n-R2 VIII
wherein R~ is chloro or bromo. The preferred acylating agents are those wherein Ra is chloro. Thus, the most highly preferred individual acylating agents are 4-[2-(piperidin-1-yl)ethoxy3benzoyl chloride, 4-[2-(hexamethyleneimin-1-yl)ethoxy]benzoyl chloride, 4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl chloride, 4-[2-(dimethyl ~m; no ) ethoxy]-benzoyl chloride, 4-[2-(diethyl ~m; no)ethoxy]benzoyl chloride, and 4-[2-(diisopropyl~m; no)ethoxy]benzoyl chloride.
The acyl chloride used as an acylating agent may be prepared from the correspon~;ng carboxylic acid by reaction with a typical chlorinating agent such as thionyl chloride.
Care must be taken to remove any excess chlorinating agent from the acyl chloride. Most conveniently, the acyl chloride is formed ;n S; tn, and the excess chlorinating agent is distilled off under vacuum.

WO9S/17095 , ; 2 ~ 7 ~ l 2 7 PCT~S94/146S5 It is generally preferred tha~ an equimolar amount of the compounds of Formula VII and VIII are reacted together. If desired, a small excess of either reactant may be added to assure the other is fully consumed. It is generally preferred to use a large excess of the acylation catalyst, such as about 2-12 moles per mole of product, preferably about S-lO moles of catalyst per mole of product.
The acylation is rapid. Economically brief reaction times, such as from about 15 minutes to a few hours provide high yields of the acylated intermediate. Longer reaction times may be used if desired, but are not usually advantageous. As usual, the use of lower reaction temperatures call for relatively longer reaction times.
The acylation step is ended and the optional demethylation step is begun by the addition of a sulfur compound selected from the group consisting of methionine and compounds of the formula xl-S _ya wherein xl is hydrogen or unbranched Cl-C4 alkyl, and ya is Cl-C4 alkyl or phenyl. The sulfur compounds are, preferably, the alkylthiols, such as methanethiol, ethanethiol, isopropanethiol, butanethiol, and the like; dialkyl sulfides, such as diethyl sulfide, ethyl propyl sulfide, butyl isopropyl sulfide, dimethyl sulfide, methyl ethyl sulfide, and the like; benzenethiol; methionine; and alkyl phenyl sulfides, such as methyl phenyl sulfide, ethyl phenyl sulfide, butyl phenyl sulfide, and the like.
It has been found that demethylation is most efficient when a substantial excess of the sulfur compound is used, in the range of about 4 to about lO moles per mole of the starting benzofuran. The process may be carried out, although less efficiently, with a smaller amount of the sulfur compound (in the range of abou~ 2 to 3 moles per mole of the starting compound). It is also possible to use a small amount of the sulfur compound, and to improve the yield WO 95tl7095 ; ; ` 2 1 7 6 1 2 7 PCT/US94114655 by the addition of about 1 to 3 moles of an alkali metal halide, such as sodium, potassium, or lithium chloride, bromide, or iodide.
The demethylation reaction goes well at about ambient temperature, in the range of from about 15-C to about 30 C, and such operation is preferred. The demethylation may be carried out, however, at temperatures in the range of from about -30 C to about 50 C if it is desired to do so. Short reaction times, in the range of about one hour, have been found to be sufficient.
After the product has been demethylated, it is recovered and isolated by conventional means. It is customary to add water to decompose the complex of the acylation catalyst. Addition of dilute aqueous acid is advantageous. The product precipitates in many instances, or may be extracted with an or~anic solvent according to conventional methods. The examples below further illustrate the isolation.
In an alternative process an intermediate compound of Formula IX

~OCH3 H3 coJ~ OCH3 IX

is synthesized by the reaction of 2-hydroxy-4-methoxybenzaldehyde and l-(4-methoxyphenyl)-2-(4-methoxyphenyl)e~h~none, essentially as described in Preparation 3a, ;n~r~. This reaction usually employs equimolar amounts of the two reactants although other ratios are operable. The reaction is performed in a non-reactive solvent such as ethyl acetate, chloroform, and the like, in the presence of an acid. Hydrochloric acid, particularly WOg5/17095 ; i ~ 2 i 7 6 ~ 2 7 pcT~ss4ll465s .

when created by bubbling anhydrous hydrogen chloride, is an especially preferred acid. Lower alkyl alcohols are usually added to the non-polar solvent so as to retain more of the hydrochloric acid created ;n situ, with ethanol and methanol being especially preferred. The reaction is performed at temperatures ranging from ambient temperature up to the reflux temperature of the mixture. This reaction results in the synthesis of a compound of Formula X

H3CO ~)~(OOCH3 or an equivalent anion if hydrochloric acid is not used, which is then oxidized to the compound of Formula IX by the lS addition of hydrogen peroxide. The intermediate of Formula X
may be isolated or may preferably be converted to the compound of Formula IX in the same reaction vessel.
The compound of Formula IX is then selectively demethylated, essentially as described in Preparation 4a, infra to yield the compound of Formula XI

Q~C~

XI

The ether of the compounds of Formula I is then produced by the substitution o~ the hydrogen on the hydroxy group by an alkyl or halide.

WO95/17095 ,. ,A ' _' 2 ~ 76 t 27 PCT/US9-1/1465 Those compounds of Formula I in which ~A~ equals -N(R~ are prepared in essen~ially the same mAnner as the substituted benzofurans described su~ra. Example 33, infra, provides one such protocol for synthesizing the substituted indoles of this invention.
Those compounds of Formula I in which UA~ equals -S ()m~ are prepared in essentially the same mAnnPr as the substituted benzofurans described $uDra. The examples infra provide several exempli~ications of these benzothiophenes and the oxidated derivatives thereof.
Those compounds of Formula I in which m is one or two may be prepared by oxidation of the corresponding benzothiophene in which m is zero. Oxidation may be carried out by treating the benzothiophene with an oxidizing agent, for example, m-chloroperbenzoic acid, or the like, for a time su~ficient to achieve formation of the sulfoxide group. The progress of the oxidation reaction may be monitored by thin layer chromatography methods.
The compounds used in the methods of this invention form pharmaceutically acceptable acid and base addition salts with a wide variety of organic and inorganic acids and bases and include the physiologically acceptable salts which are often used in pharmaceutical chemistry. Such salts are also part of this invention. Typical inorganic acids used to form such salts include hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric and the like.
Salts derived from organic acids, such as aliphatic mono and dicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, may also be used.
Such pharmaceutically acceptable salts thus include acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, ~-hydroxybutyrate, butyne-1,4-dicarboxylate, hexyne-1,4-dicarboxylate, caprate, caprylate, cinnAmAte~ citrate, formate, fumarate, glycollate, heptanoate, hippurate, hydrochloride, lactate, malate, maleate, hydroxymaleate, ~alonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate, oxalate, phthalate, teraphthalate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, propiolate, propionate, phenylpropionate, salicylate, sebacate, succinate, suberate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzene-sulfonate, p-bromobenzenesulfonate, chlorobenzenesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate, naphthalene-l-sulfonate, naphthalene-2-sulfonate, p-toluenesulfonate, xylenesulfonate, tartarate, and the like. A preferable salt is the hydrochloride salt.
The pharmaceutically acceptable acid addition salts are typically formed by reacting a compound of Formula I with an equimolar or excess amount of acid. The reactants are generally combined in a mutual solvent such as diethyl ether or benzene. The salt normally precipitates out of solution within about one hour to 10 days and can be isolated by filtration or the solvent can be stripped o~f by conventional means.
Bases commonly used for formation of salts include ammonium hydroxide and alkali and alkaline earth metal hydroxides and carbonates, as well as aliphatic and aromatic amines, aliphatic ~i~m;nes and hydroxy alkyl~m;ne~. Bases especially useful in the preparation of addition salts include Amm~nium hydroxide, potassium carbonate, calcium hydroxide, methyl~m;ne, diethylamine, ethylene ~;Am;ne~
cyclohexyl~m;np and ethanolamine.
The pharmaceutically acceptable salts frequently have ~nh~nced solubility characteristics compared to the compound from which they are derived, and thus are often more ~m~n~hle to formulation as liquids or emulsions.

-WO 95/17095 ~ , ,, 2 ~ 7 6 1 2 7 PCTtUS94/14655 ~xAmnles The following experiments illustrate the preparation of the benzofurans, benzothiophenes and indoles employed in the present invention. The terms "NMR", "IR" or ~MS~ following a synthesis protocol indicates that the nuclear magnetic resonance spectrum, infrared spectrum, or the mass spectrometry was performed and was consistent with the title product.
Pre~ArAt;on 1 A

Synthesis of 2-(3-methoxyphenoxy)-1-(4-methoxyphenyl)ethAno~e.
H3C~O O~OCH3 In a one liter round-bottom flask, fitted with a condenser and nitrogen inlet, were added 3-methoxyphenol (12.4 g, 0.1 mole), 4-methoxyphenacyl bromide (22.9 g, 0.1 mole), potassium carbonate (17.3 g, 0.125 mole) in 100 ml of 2-butanone. This mixture was heated to 80 C and was maintained at this temperature for about four hours. The progress of the reaction was monitored by thin layer chromatography (silica gel, 9:1 toluene:ethyl acetate).
After the four hours at 80 C the reaction mixture was cooled and the reaction mixture was partitioned by the addition of water. The organic phase was L~ ved and the a~ueous layer was washed with 2-but~none. The organic layers were then combined, dried over ~gn~sium sulfate, and the solvents were removed in vacuo to yield 31.1 grams of a yellow oil. The yellow oil was further purified by chromatography, the fractions contA;ning the desired product were then crystallized. A11 of the crystAll;ne fractions WO 95/1709~ 2 ~ 7 6 1 2 7 PCT/US94/14655 -- 41 ~

were combined and then dissolved in 80 ml of hot ethanol.
Fifteen milliliters of hot water was then added, the product was crystallized, and subsequently washed with an ethanol/water mixture to yield 19.1 g (70%) of the desired title product. mp 52.5 -53.5 C.
AnalysiS for Cl6Hl64:
Theory: C, 68.08; H, 5.71; N, 2.84.
Found: C, 67.86; H, 5.51; N, 2.88.

PreD~r~t;on 2~

Synthesis of 2-methoxyphenyl-6-methoxybenzofuran.

1{3Co OCH~
The cyclization of the product of Preparation la was performed essentially as described in C. Goldenberg, h;mi e Ther~el~ti~ue, 398-411 tl973). In a 500 ml 3-neck round bottom flask polyphosphoric acid (30 g) was added to 200 ml of xylene. The mixture was then heated to about 120'C To this heated mixture was then added 2-(3-methoxyphenoxy)-l-(4-methoxyphenyl)ethanone (10 g, 0.037 mole), prepared as described sll~ra, and the temperature was raised to about 170-C, and maintained at that temperature for about eight hours. The reaction mixture was then cooled and water was added.
The dark aqueous layer was separated from the yellow organic phase. The organics were washed with waterand by aqueous sodium carbonate, and then dried over anhydrous magensium sulfate. The solvents were removed in vacuo, resulting in a yellow-orange solid. The product was recrystallized from a m;nimllm of hot acetone, followed by the addition of ethanol and wa~er. The residual acetone was 2 t 7 7 PC~Sg~/146~5 WO95/17095 ~ If U~ L 7 removed by boiling. Cooling to room temperature yielded white crystals (2.09 g, 22% yield). mp 158-C.
Analysis for C16H143: -Theory: C, 75.58; H, 5.55; O, 18.88.
Found: C, 75.33; H, 5.67; 0, 18.62.

Pre~ r~ t ion 3~

Synthesis of 2-(4-methoxyphenyl)-3-(4-methoxybenzoyl~-6-methoxybenzofuran ~OCH3 ~C
~_~ OCH3 In a 250 ml 3-neck round bottom flask were added 2-hydroxy-4-methoxybPn~ phyde (10 g, 65.7 mmol), 1-(4-methoxyphenyl)-2-(4-methoxyphenyl)eth~none (16 g, 62.6 mmol), ethyl acetate (100 ml~ and ethanol (25 ml). The reaction mixture was then warmed to about 45 C until all the starting ma~erials were dissolved. Hydrogen chloride gas was then bubbled in for about 30 minutes, resulting in the ormation of a bright red coloration. The reaction was then allowed to stand at room temperature for about two hours at which time the solvents were lel,.o-ved in vacuo to leave a bright red oil.
The red oil was dissolved in 180 ml of methanol and 30 ml of 20% sulfuric acid was added with stirring and cooling. Hydrogen peroxide (30 ml) was added dropwise and the ~;xt~re was allowed to stir for about 30 minutes. A
saturated sodium chloride solution (500 ml) and ethyl acetate (300 ml) were added to the reaction mixture and the organic fraction was r~.ov~d. The organic layer was washed with a saturated sodium chloride solution, dried, and the solvents were removed in vacuo to provide 25 g of a reddish brown oil WO 95/17095 ; - ; ~ PCT/US94/14655 ~ 1 7 ~

which was further purified by chromatography to yield the title product (1.25 g) as a yellow oil. mp 106-lO9 C.
Analysis for C24H205:
~heory: C, 74.21; H, 5.19; O, 20.60.
Found: C, 74.07; H, 5.22; O, 20.38.

Pre~r~tion 4~

Synthesis of 2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxybenzofuran C
~ OCH3 In a three-neck round bottom flask under a nitrogen atmosphere and cooled in an ice bath, ethanethiol (0.95 ml, 1.288 mmol) was dissolved in 10 ml of anhydrous N,N-dimethylformamide. To this solution was added n-butyllithium (0.60 ml of a 1.6 M in hexane solution, 0.966 mmole) followed by the addition of 2-(4-methoxyphenyl)-3-(4-methoxybenzoyl)-

6-methoxybenzofuran (250 mg, 0.644 mmole), prepared as described in Preparation 3, sllnr~. The reaction mixture was then heated to 80 C and allowed to remain at that tempeature for about 16 hours.
The reaction mixture was then poured into 1 N
hydrochloric acid and extracted with ethyl acetate. The organic layer was then washed with a saturated sodium chloride solution, dried over magnesium sulfate, filtered and the solvents were removed in vacuo. The desired product was further purified by column chromatography. The product was then crystallized from methanol yielding 130 mg (81%) of the desired product. mp 148-149 C.
Analysis for C23Hl8os:

WO95/17095 ~ 76 1 27 PCTIUS94/1465~ ~
., Theory: C, 73.79; H, 4.85, O, 21.37.
Found: C, 73.68; H, 5.12; o, 21.17.

FX~mnle 18 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran ~ O--CH2 ~ CH2 - N~
O~J

H3COJ~OCH3 Method A: Acylation of Benzofuran 4-[2-(Piperidin-l-yl)ethoxy]benzoyl chloride (0.562 g, 1.96 mmol) was added to ethylene chloride (20 ml), followed by the addition of 2-methoxyphenyl-6-methoxybenzofuran (0.500 g, 1.96 mrnol), prepared as described in Preparation 2a, suDra. This mixture was stirred at room temperature as alllminllm trichloride (1.96 g, 14.7 mmol) was added. This reaction mixture was then stirred overnight.
The reaction mixture was then poured over ice, and extracted with warm chloroform (3 x 50 ml). The chloroform was removed by evaporation. Sodium carbonate, water and ethyl acetate were then added and the organic layer was ~einoved, ~fied over magnesium sulfate, and the solvents were removed in vacuo to provide a yellow oil. The desiredproduct was further purified by chromatography of the yellow oil to yield the desired title product.
NMR, IR, MS.

wo 95/17095 ~ 7 6 ~ 2 7 PCT/USg4/l4655 Anaiysis for C30H31NOs:
Theory: C, 74.21; H, 6.44; N, 2.88; O, 16.47.
Found: C, 74.11; H, 6.71; N, 2.75; O, 16.57.

5 Method B: Alkylation of 2-(4-mechoxyphenyl)-3-(4-hydroxybenzoyl)-6-me~hoxybenzofuran.

In 100 ml of anhydrous N,N-dimethylformamide in a 500 ml round bottom flask were added 2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxybenzofuran (10.50 g, 28 mmol), prepared as described in Preparation 4a, su~r~, and potassium carbonate (6.20 g, 34 mmol). This mixture was heated to lOO C and then 2-(piperidin-1-yl)ethyl chloride (6.20 g, 34 mmol) was added gradually. The reaction mixture was kept at 15 lOO'C for about one hour.
The N,N-dimethylformamide was evaporated and the residue was dissol~red in ethyl acetate and water. The ethyl acetate layer was removed and the aqueous layer was washed with more ethyl acetate. The organic fractions were 20 combined, dried over magnesium sulfate, and the solvents were removed in vacuo, yielding 13.3 g of a yellow oil which crystallized upon st~nfli ng . The product was recrystallized from methanol cooled to -30 C prior to filtra~ion, yielding 11.4 g (84%) of the desired product as pale yellow cr~rstals.
mp 87-89 C.
AnalysiS for C3oH3lNos:
Theory: C, 74.21;H, 6.44; N, 2.88; O, 16.47.
Found: C, 74.31;H, 6.34; N, 2.63; O, 16.47.

3 o F.x~m~l e 1 9 SynthesiS of 2-(4-hydroxyphenyl)-3-[4-t2-(piperidin-1-yl)ethoxy3benzoyl]-6-hydroxybenzo~uran WO9S/17095 ;- 2 ~ 76 1 27 PCT~Sg4/1465~ ~

~ O-CH,-cH~-N ~
o~l HO~

The title product was prepared by the demethylation of 2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran, the product of Example la, $u~ra. In a 250 ml three-neck round bottom flask were combined ethylene chloride (50 ml) and alllminllm trichloride (9.60 g, 72 mmol) and ethanethiol (6.39 g, 103 mmol) to create a pale yellQw liquid. To this liquid was then added the product of Example la (5.00 g, 10.3 mmol) in a gradual fashion. A red oil precipitated and the mixture was stirred for about 20 minutes. After cooling the reaction mixture in an ice bath 100 ml of tetrahydrofuran was added and the mixture was allowed to stir until all of the oil had gone into solution.
The reaction mixture was then poured over ice (200 ml) and water (500 ml) and concentrated hydrochloric acid (10 ml) were added. The oil which precipitated was separated from the liquid by decantation. The liquid was extracted with chloroform (warm, 2 x 300 ml). The oil was dissolved by mixing with ethyl acetate, chloroform, sodium bicarbonate, and a small amount of sodium hydroxide. The chloroform extract and the dissolved oil were transferred to separatory funnel and washed with sodium bicarbonate. The organic phase was then dried over magnesium sulfate and the solvents were ~e,.,oved by evaporation to yield a yellow foam, which was further purified by high performance liquid chromatography.
NMR, IR, MS.

95/17095 ? 1 7 6 ! 2 7 PcT~ss4/14655 Analysis for C2gH27Nos:
Theory: C, 73.51; H, 5.95; N, 3.06.
Found: C, 70.45; H, 6.34; N, 4.02.

~xAmnle 20 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran hydrochloride ~ o-CH2 CH2-0~1 H3CO ~ OCH~

The title compound is prepared essentially as described in the process for preparing the compound of Example 18 except that 4-[2-(pyrrolidin-1-yl)ethoxy3benzoyl chloride is employed in the synthesis of Method A in place of 4-[2-(piperidin-1-yl)ethoxy]benzoyl chloride or 2-(pyrrolidin-l-yl)ethyl chloride is employed in the synthesis of Method B in place of the 2-(piperidin-1-yl)ethyl chloride.
FxAmnle ~1 Synthesis of 2-(4-hydroxyphenyl)-3-~4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-6-hydroxybenzofuran hydrochloride WO95117095 ~ PcT~S94114655 _ ~ o-CH, cH2-HO ~ HCl The title compound is prepared essentially as described in Example l9 except that 2-(4-methoxyphenyl)-3-[4-[2-(pyrrolidin-l-yl)ethoxy]benzoyl]-6-methoxybenzofuran is used as the startiny material instead of 2-(4-methoxyphenyl)-3-[4-[2-(piperidin-l-yl)ethoxy]benzoyl]-6-methoxybenzofuran.
NMR, IR, MS.
Analysis for C27H26NOsCl:
Theory: C, 67.57; H, 5.46; N, 2.92.
Found: C, 67.84; H, 5.56; N, 2.87.

~mnle 22 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(diethyl~mino)ethoxy]benzoyl]-6-methoxybenzofuran ~CH2-CH3 o CH2CH2-N~
o9~

H~C~ ~ OCH~

The title compound was prepared by reacting the compound of Preparacion 4a sl7~rr7, 2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxybenzofuran (l0 g, 26.7 mmol) which WO 95/17095 ,,~ . i 2 1 7 6 ~ 2 7 PCT/US94/14655 is dissolved in 200 ml of N,N-dime~hylformamide with an e~uimolar amount of 2-(N,N-diethyl~mino)ethyl chloride (6.4 g, 32 mmol) and potassium carbonate (11.06 g, 80.2 mmol).
The mixture was heated to 100-C and was maintained at that temperature for about two hours. The reaction mixture was then cooled to room temperature and maintained at this temperature overnight while stirring.
The solven~s were then removed by evaporation and th~ residue was extracted from wa~er with ethyl acetate and washed twice with a saturated sodium chloride solution. The organic phase was dried over sodium sufate and the solvents were removed in vacuo. The material was crystallized from hexane and recrystallized in methanol.
NMR, IR, MS.5 Analysis for C29H31NO5:
Theory: C, 73.55; H, 6.60; N, 2.96.
Found: C, 73.29; H, 6.50; N, 2.84.
Fx~mn l e 2 3 synthesis of 2-~4-hydroxyphenyl)-3-[4-t2-( diethyl ~m i no ) ethoxy]benzoyl]-6-hydroxybenzofuran hydrochloride ~CH2 CH3 OcH

~ HCl HO ~ OH

The title compound was prepared essentially as described in Example 19, sl~nr~, except that the compound of Example 5, 2-(4-methoxyphenyl)-3-[4-[2-WO95/1709~ . 2 1 76 1 ~7 PCT/US94/14655 ~

(diethylamino)ethoxy]benzoyl]-6-methoxybenzofuran, was used as the starting material to be demethylated.
NMR, IR, MS.
Analysis for C27~28NO5Cl:
Theory: C, 67.29; H, 5.86; N, 2.91.
Found: C, 67.54; H, 5.64; N, 2.92.

Fx~mnle 24 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(diiso~ulu~yl~m;no)e~hoxy]benzoyl]-6-methoxybenzofuran CH_C
O-CH2CH2 N~
, CH-CH3 O ~ ~ CH3 15H3CO ~ OCH~
The title compound was prepared by reacting the compound of Preparation 4a ~nnr~, 2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxybenzofuran (10 g, 26.7 mmol) which is dissolved in 200 ml of N,N-dimethylformamide with 2-(N,N-diiso~Lo,uyl~mino)etllyl chloride (6.4 g, 32 mmol) andpotassium carbonate (11.06g, 80.2 mmol). The mixture was heated to 100-C and was maint~;ne~ at that temperature for about two hours. The reaction mixture was then cooled to room temperature and maint~;ne~ at this temperature overnight while stirring.
The solvents were then removed by evaporation and the residue was extracted from water with ethyl acetate and washed twice with a saturated sodium chloride solution. The organic phase was dried over sodium sufate and the solvents ~ WO95/17095 ~ ~ ; 2 1 7 6 1 2 7 PCT~S94/14655 were removed in vacuo. The material was crystallized from hexane and recrystallized in methanol.
NMR, IR, MS.
Anaiysis for C33H3gNOs:
Theory: C, 74.83; H, 7.42; N, 2.64.
Found: C, 74.68; H, 7.14; N, 2.76.

F.x~mr~ 1 e ~ ';

Synthesis of 2-(4-hydroxyphenyl)-3-[4-[2-(diisopropylamino)ethoxy]benzoyl]-6-hydroxybenzofuran hydrochloride CH-CH~
~0 CH 2-CH 2 - N

o ~ CH3 / HCl lS HO ~

The title compound was prepared essentially as described in Example l9, ~a, except that the compound of Example 24, 2-(4-methoxyphenyl)-3-[4-[2-(diisopropylamino)ethoxy]benzoyl]-6-methoxy~enzofuran, was used as the starting material to be demethylated.
NMR, IR, MS.
Analysis for C29H32NOsCl:
Theory: C, 68.29; H, 6.32; N, 2.75.
Found: C, 68.53; H, 6.49; N, 2.74.

:
WO95/17095 2 l 7 6 t 2 7 PCT~Sg~/1465~ ~

~x~le 26 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(dimethyl A mln n )ethoxy]benzoyl]-6-methoxybenzofuran ~ O-CH,CH2-H3CO ~ OCH3 The title compound was prepared essentially as described in Example 24, supra, except that 2-(N,N-dimethylamino)ethyl chloride was reacted with 2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxybenzofuran instead of the 2-(N,N-diisopropyl~mino)ethyl chloride employed in that example.
NMR, IR, MS.
Analysis for C27~27NOs:
Theory: C, 72.79; H, 6.11; N, 3.14.
Found: C, 72.51; H, 6.27; N, 3.10.

Fx~mnle 27 Synthesis of 2-(4-hydroxyphenyl)-3-[4-t2-(dimethyl~m~no)ethoxy]benzoyl]-6-hydroxybenzofuran WO9S/17095 = ;. = - -` i '' 2 t 76 t 27 PCT/US91/146S~i -CH2-CH~-N ~

9~

~ OH

The title compound was prepared essentially as described in Example 19, sll~r~, except that the compound of Example 26, 2-(4-methoxyphenyl)-3-[4-[2-(dimethylamino)ethoxy]benzoyl]-6-methoxybenzofuran, was used as the starting material to be demethylated.
NMR, IR, MS.
Analysis for C2sH23NOs:
Theory: C, 71.93; H, 5.55; N, 3.36.
Found: C, 70.69; H, 5.51; N, 3.16.

~.~mn l e 28 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(hexamethyleneimin-l-yl)ethoxy]benzoyl]-6-methoxybenzofuran 0-CH2-CH2--N~ ) o~

H3Co~oc~3 The title compound was prepared essentially as described in Example 24, supra, except that 2-thexamethyl~ne;m;n-l-yl)ethyl chloride was reacted with 2-(4-WO95/17095 ~ 2 ~ 76 1 27 PCT/US9~/14655 ~
. . .

methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxybenzofuran instead of the 2-(N,N-diisopropylamino)ethyl chloride employed in that example.
NMR, IR, MS.
AnalysiS for C3lH33NOs:
Theory: C, 74.53; H, 6.66; N, 2.80.
Found: C, 74.69; H, 6.70; N, 2.75.

~x~mnl e 29 Synthesis of 2-(4-hydroxyphenyl)-3-[4-[2-(hexamethyleneimin-l-yl)ethoxy]benzoyl]-6-hydroxybenzofuran hydrochloride ~, o- CH2- CH2--N~

HCl J~ OH

The title compound was prepared essentially as described in Example 19, sllnr~, except that the compound of Example 28, 2-(4-methoxyphenyl)-3-[4-[2-~hexamethyleneimin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran, was used as the starting material to be demethylated.
NMR, IR, MS
Analysis for C2gH30ClNOs:
Theory: C, 68.57; H, 5.95; N, 2.76.
Found: C, 67.28; H, 6.13; N, 2.66.

~ 21~6127 ~x~m~le 30 Synthesis of 2 - ( 4-hydroxyphenyl)-3-[4-~2-(piperidin-1-5 yl)ethoxy]benzoyl]-6-hydroxybenzofuran hydrochloride ~0-CH2-CH2 N~
0~~1 HCl HO~

The title compound was prepared ~y dissolving the compound of Example 19, 2- (4-hydroxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-hydroxybenzofuran, (3.1 g, 6.8 mmol) in 15 ml of methanol and treating with an excess of 3% hydrochloric acid in methanol. The volume was then reduced by boiling to 15 ml. Warm water (20 ml) was then 15 added and the reaction mixture was further warmed to clarify.
The reaction mixture was then filtered, followed by gradual cooling to O'C, at which temperature the mixture was maintained for about one hour. The crystals, which had precipitated, were collected by filtration and washed with 20 cold water. The pale yellow c~ystals were dried overnight, resulting in 2.82 g (84%) of the desired title product. mp 213-215-C.
NMR, IR, MS.
Analysis for C2gH2gNOsCl:
Theo~: C, 68.08; H, 5.71; N, 2.84; O, 16.19.
Found: C, 67.86; H, 5.51; N, 2.88; O, 15.93.

!~ .. , : 2 1 7 6 1 2 7 ~.x~mn1 e 31 Svnthesis of 2-(4-hydroxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzofuran hydrochloride --CH2cH2-HCl ~ OH

The 2-(4-hydroxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzofuran was prepared essentially as described in Example 19, except that phenol was used as a starting material in the synthesis described in Preparation 2a instead of 3-methoxy phenol. The hydrochloride salt of this substituted benzofuran was prepared essentially as described in Example 30, supra.
NMR, IR, MS.
Analysis for C2gH2gNO4Cl:
Theory: C, 70.36; H, 5.91; N, 2.93.
Found: C, 70.46; H, 5.84; N, 2.84.
~mnl e 32 Synthesis of 2-phenyl-3-[4-[2-(piperidin-1-2S yl)ethoxy~benzoyl]-6-hydroxybenzofuran hydrochloride WO 95/17095 ~ 7 ~ 1 ~ 7 PCT/US94/14655 O- CH~CH2 -N~

~J HC l H0~

The 2-phenyl-3-[4-[2-(piperidin-1-yl~ethoxy]benzoyl]-6-hydroxybenzofuran was prepared essentially as described in Example 19, except that phenacylbromide (also known as a-bromoacetophenone) was used as a starting material in the synthesis described in Preparation la instead of 4-methoxyphenacylbromide. The hydrochloride salt of this substituted benzofuran was prepared essentially as described in Example 30, ~llnr~.
NMR, IR, MS.
Analysis for C2gH2gNO4Cl:
Theory: C, 70.36; H, 5.90; N, 2.93.
Found: C, 70.39; H, 6.01; N, 2.91.

~x~mnl e 33 Synthesis of 1-ethyl-2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-hydroxyindole hydrochloride salt , WO 9S/17095 . ~ 2 ~ 7 6 1 2 7 PCT/US94/14655 N~
~O--CH2~CH --~ .HCl HO ~ OH

To 814 milliliters of concentrated hydrochloric acid in a 3 liter, 3-neck round bottom flask which had been cooled to 0 C was added 3-methoxyaniline (99.26 g, 0.806 mole). Sodium nitrate (55.61 g, 0.806 mole), dissolved in 249 milliliters of water, was added dropwise to the 3-methoxyaniline solution at such a rate that the reaction temperature never exceeded 0 C. This mixture was then stirred for about 90 minutes.
Stannous chloride (545.57 g, 2.418 mol), dissolved in 497 milliliters of concentrated hydrochloric acid, was added dropwise to the reaction mixture at such a rate that the reaction temperature never exceeded 5'C. This mixture was then stirred for about two hours after the addition of the stannous chloride was completed, resulting in the formation of a thick, beige, chalky emulsion. The solid was removed by filtration, stored overnight in one liter of water and then basified with a 25~ solution of sodium hydroxide.
This aqueous solution was extracted with diethyl ether (3 x 1 liter) and then dried over sodium sulfate. The solvents were removed in vacuo, resulting in a brown oil of 3-methoxyphenylhydrazine (76.3 g, 69% yield).
The 3-methoxyphenylhydrazine (76.3 g, 0.~52 mole) prepared sllnra, was dissolved in 400 milliliters of ethanol.
To this mixture was added p-methoxyacetophenone (82.80 g, 0.552 mole) followed by the addition of about 6 drops of WO95117095 ~ 2 1 7~ ~ 2~ PCT/US94/1465S

hydrochlorlc acid. This mlxture was then stirred for about seven hours under a nitrogen atmosphere, followed bv storage at 4 C for about 3 days.
The white solid was then removed from the suspension by filtration under vacuum and then dried L~
vacuo, resulting in 135.2 grams (91% yield) of ~(3-methoxyphenyl)hydrazono]-l-methyl-4-methoxybenzylidene of the following formula as a pale gray solid.

H3 COJ~ N--N= C~ OCH3 Zinc chloride (66.5 g, 0.49 mole) was added to a 3-neck round bottom flask under a nitrogen atmosphere. The flask and its contents were then heated to 200 C at which time the hydrazone (26.4 g, 0.098 mole) prepared ~nl~ was added. The mixture was stirred for about 17 minutes, resulting in the formation of a brown tar and the evolution of some gas. The brown tar was then poured into two liters of 0.075 N hydrochloric acid and this mixture was stirred for about 48 hours, resulting in the formation of a yellow solid.
The solids were removed by filtration and were then recrystallized from methanol. The solids were again removed by filtration and the solvents were removed ;n V~CUO to yield the desired 2-(4-methoxyphenyl)-6-methoxyindole (5.50 g, 22 yield) as a white cryst~ e product.
The 2-(4-methoxyphenyl)-6-methoxyindole (2.0 g, 8 mmol) was dissolved in 40 milliliters of N,N-dimethylformamide. This solution was added dropwise to a solution of sodium hydride (O.48 g, 12 mmol) in ten milliliters of N,N-dimethylformamide. This reaction mixture was then stirred at room temperature for 1 hour at which time a solution of ethyl iodide (1.9 g, 12 mmol) in N,N-dimethylformamide (10 ml) was added dropwise over five WO95117095 . ; ~ 2 ~ 76 1 27 PCT~S9~/1465~

minutes. This mixture was then stirred at room temperature for about two hours.
The reac~ion was quenched by the addition of methanol. The volume of the solvents was reduced by vacuum, leaving a brown oil. This oil was diluted with chloroform, washed with 5 N sodium hydroxide (3 x 75 ml), followed by washing with water (2 x 200 ml). The organic layer was dried over sodium sulfate and the solvents were removed in v~cuo leaving 2.3 g of the desired intermediate 1-ethyl-2-(4-methoxyphenyl)-6-methoxyindole as white crystals.
The preceding intermediate was acylated at the 3-position by first placing N,N-dimethyl-4-methoxybenzamide (1.43 g, 8 mmol), in a 100 ml flask cooled to O C. To this was then added phosphorous oxychloride (6.1 g, 40 mmol) dropwise at such a rate that the reaction temperature never exceeded 20 C. The reaction mixture was allowed to warm to room temperature and was stirred for about 30 minutes. The reaction mixture was then cooled to O C and the l-ethyl-2-(4-methoxyphenyl)-6-methoxyindole (1.5 g, 5.33 mmol) prepared $~ra, was added and the reaction mixture was then heated ~o 75 C and maint~ineA at this temperature for about three hours.
A~ter this incubation, the reaction mixture was poured over ice and diluted with water. The layers were separated and the organic phase was washed with water (150 ml). The organic layer was dried over sodium sulfate and ~he oslvents were removed ; n V~Cllo to yield a dark brown/black oil. This oil was taken up in 50 milliliters of methanol and cooled to O C. Thi.s solution was then basified by the dropwise addition of 2N sodium hydroxide (50 ml). The mixture was then heated to reflux for about 5 minutes, then cooled overnight at ~'C.
The precipitate was then removed by filtration and recrystallized from methanol, resulting in 2.21 grams (86%
yield) of the intermediate 1-ethyl-2-(4-methoxyphenyl)-3-(4-methoxybenzoyl)-6-methoxyindole as a yellow precipitate.

WO95/17095 ~ ; . '-; ~` 2 1 76 1 2;7 PCT/US94/1465 The above lntermedia~e (2.1 g, 5.05 mmol) was then admixed with sodium thioethoxide (0.85 g, 10.11 mmol) in N,N-dimethylformamide (12 ml). The reaction mixture was then heated to 85 C and maintained at this temperature for about six hours. The desired intermediate l-ethyl-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)-6-methoxyindole was then recrystallized from ethyl acetate.
This intermediate (1.5 g, 3.74 mmol) was then reacted with 2-(piperidin-1-yl)ethyl chloride hydrochloride (1.38 g, 7.5 mmol) in N,N-dimethylformamide (60 ml) in the presence of cesium carbonate (3.26 g, 10 mmol). This admixture was heated to 80 C and maintained at this temperature for about two hours.
The precipitate was collected by filtration and then taken up in chloroform, and washed with 2 N sodium hydroxide (3 x 125 ml) and water (3 x 100 ml). The organic fraction was then dried over sodium sulfate and the solvents were removed ;n v~c-lo to yield 2.05 grams (95% yield) of 1-ethyl-2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxyindole as a gray foam.
This intermediate (1.0 g, 1.82 mmol) was dissolved in dichloromethane (10 ml) and cooled to O'C. To this mixture was then added the Lewis acid alllmlnllm chloride (1.2 g, 9 mmol) and the reaction mixture was then stirred for five minutes. Ethanol (3 ml) were then added and the reaction mixture was stirred on ice for about 15 minutes. The temperature of the reaction mixture was slowly raised to reflux and maintained at reflux for about 1.5 hours.
The reaction mixture was then cooled to O'C and this temperature was maintained as tetrahydrofuran (5 ml) was added. To this mixture was then added 20% hydrochloric acid in water (5 ml) and the reaction mixture was cooled back to O C at which time five milliliters of water was then added, resulting in the formation of a yellow gum. This suspension was then placed at -40 C and kept at this temperature for about 48 hours, after which time a grayish material was removed from the mixture by filtration. Thin layer WO95/l7095 2 1 7 6 1 2 7 PCT~S9~/146~5 chromatography confirmed this precipitate as the desired title product.
NMR, MS.
AnalySis for C30H33ClN2O4:
Theory: C, 69.15; H, 6.38; N, 5.38.
Found: C, 69.09; H, 6.43; N, 5.53.

~x~mnl e 34 Synthesis of 2-(4-hydroxyphenyl)-3-[4-t3-(piperidin-l-yl)propoxy]benzoyl~-6-hydroxybenzo[b]thiophene hydrochloride ~ocH2cH2cH2--N3 ~C

HOJ~ OH
The title compound was prepared essentially as described in U.S. Patent 4,380,635, which is herein incorporated by reference with the exception that 4-[3-(piperidin-1-yl)propoxy]benzoyl chloride was used to acylate the substituted benzo[b]thiophene rather than the 4-[2-(piperidin-1-yl)ethoxy]benzoyl chloride employed therein.

~xAmnle 35 Synthesis of 2-phenyl-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]benzo~b]thiophene WO 95/1709S ~ PCT/US94/1465!i ~ 2176127 OCH7CH,--N

~c~

The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.~m~ l e 36 Synthesis of 2-phenyl-3-[4-~2-(pyrrolidin-1-yl)ethoxy3benzoyl~-6-methoxybenzo[b]thiophene citrate o~ ~OCH2CH2--N~

HO~{~

The title compound was prepared as described in 15 u.s. Patent 4,133,814, which is herein incorporated by reference.

~x~mn l ~ 37 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene citrate WO 9S/17095 . 2 1 7 6 1 ~ 7 PCT/US94/14655 ~OCH2CH2--N~

~C
~ OCH3 The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.X~mnle 38 Synthesis of 2-(4-ethoxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-6-methoxybenzo[b]thiophene citrate ~ C OCH2 CH2--N~

HO~ OCH2 CH3 The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

~.x~mnl e 39 Synthesis of 2-(4-acetoxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl~benzo[b]thiophene citrate ~ OCH2 CH2 - N~

~C
~_ o' ` CH3 The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

~xAmnle 40 Synthesis of 2-phenyl-3-[4-~2-(piperidin-1-yl)ethoxy3~enzoyl~benzo[b]thiophene ~OCH2CH2--N~

~C
~ .' The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.~mnl e 41 Synthesis of 2-phenyl-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxybenzo[b]thiophene citrate WO95/17095 ~= ~. PcT~Sg~/14655 2 1 7 6 ~ 2 7 O~ ~ ocH2cH2-HO ~

The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.~mnle 4~

Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene citrate ~OCH2CH2--N~>

~C
~ OCH3 The title compound was prepared as described in u.S. Patent 4,133,814, which is herein incorporated by reference.

F.x~mn 1 e 43 SynthesiS of 2-(4-ethoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxybenzo[b]thiophene citrate WO 95/1709~ , , 2 1 ~ 7 2 7 PCT/US94/146SS

~OCH2CH2--N~

~C

HO f ~ OCH2 CH3 The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.~mnle 44 Synthesis of 2-(4-acetoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxyibenzoyl]benzo[b]thiophene citrate e c~OCH2 CH2 - N~>

~_ ..

The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.x~mnle 45 Synthesis of 2-(4-pentanoylphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene citrate WO 95/17095 ~ 2 1 7 6 t 2 7 PCT/US94/14655 ~OCH7CH2--N~_~

~C o ~_ ~ C~ CH3 The title compound, also known as 2-(4-valerylphenyl)-3-[4-[2-~piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene citrate, was prepared asdescribed in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.x~mnle 46 Synthesis of 2-(4-chlorophenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene citrate r~
~OCH2CH2--N~,) ~C
~-Cl The title co...yound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

~.x~mnl e 47 Synthesis of 2-phenyl-3-[4-[2-(hexamethyl~nei m i n-1-yl)ethoxy]benzoyl]benzo[b]thiophene WO95/17095 ~ . ~ 2 1 7 6 1 ~ 7 PCT~Sg4/146S5 ~ OCH2CH2-N ~

~c The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.x~m~ 1 e 48 Synthesis of 2-phenyl-3-[4-E2-(hexamethyleneimin-1-yl)ethoxy]benzoyl]-6-methoxybenzo[b]thiophene citrate ~ocH2cH2--N~

~C

HO~

The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.x~mnle 49 20 Synthesis of 2-(4-methoxyphenyl)-3-[4-[2-(hexamethylen~;min-l-yl)ethoxy]benzoyl]benzo[b]thiophene citrate WO95/17095 ~ 2 1 76 1 ~7 PCT/US94/14655 ~

OCH2 CH2--N~
~ OCH3 The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

~xAmnle 50 Synthesis of 2-(4-ethoxyphenyl)-3-~4-[2-(hexamethyleneimin-1-yl)ethoxy]benzoyl]-6-methoxybenzo~b]thiophene citrate ~OCH2CH2--N~J

~C

HO~} OCH2CH3 The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

~xAmnle 51 Synthesis of 2-(4-acetoxyphenyl)-3-[4-[2-(hexam.ethylene;min-1-yl)ethoxy]benzoyl]benzo[b]thiophene citrate WO95/17095 '~ ~ . ` . . 2 1 76 ~ 27 PCT/US94/14655 N~
~OCH2 CH,--0~ ' ~

The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

F.x~mnle 52 Synthesis of 2 - ( 4-pentanoylphenyl)-3-[4-[2-10 (hexamethyleneim;n-l-yl)ethoxy]benzoyl~benzo~b~thiophene citrate ~OCH2CH2--N~J

~C o ~_o~C ~CH3 The title compound, also known as 2-~4-valerylphenyl)-3-[4-[2-(hexamethyl~n~;m;n-1-yl)ethoxy]benzoyl]benzo~b]thiophene citrate, was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

WO 95/17095 ; . ~ - ' 2 ~ 76 1 2 7 PCT/US94/1465S

F.x~ mn 1 e 53 Synthesis of 2-(4-chlorophenyl)-3-~4-[2-(hexamethyleneimin-1-yl)ethoxy3benzoyl]benzo~b]thiophene citrate ~OCH2CH2--N~J

~C
~}Cl The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference. .

F.x~ mn 1 e 54 Synthesis of 2-(4-chlorophenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene-1-oxide /~
~ OCH2 CH2--N~_~

~C
~;~Cl The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

f ~ ~

-- 73 ~

~x;~mnle 55 Synthesis of 2-(4-chlorophenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy3benzoyl]benzo[b]thiophene-1-oxide ~OCH~CH2--N~

~}Cl The title compound was prepared as described in U.S. Patent 4,133,814, which is herein incorporated by reference.

Those compounds employed in the methods of the instant invention in which R or Rl are -OSO2-(C1-clo alkyl) or -oCN-R3 H were made essentially as described in European Patent Application 617,030, published September 28, 1994.
Those compounds employed in the methods of the instant invention wherein at least one of R1 and R is -OSo2-(cl-clo alkyl) were generally prepared by reacting a compound of Formula II

~ O-CH2CH2-X-R2 0~

HO~S{~OH II

WO95/17095 ~ ~ 76 ~ 27 PCT~Sg4/146 with an alkyl sulfonyl of Formula IIa xl-o- -(Ci-C10 alkyl) ~) .
IIa where xl is a leaving group, preferably a chloro or bromo group. This reaction is usually performed in a basic environment in the presence of a coupling catalyst such as 4-dimethylaminopyridine (DMAP). Most preferred solventsinclude the lower alkyl amines, especially triethyl ~mi ne .
While this thioester formation reaction may be performed at equal molar ratios of the two reactants, it is usually preferred to employ a 2-3 molar excess of the alkyl sulfonyl compound so as to complete the reaction.

The following examples will illustrate preparation of these compounds of this invention but are not intended to limit it in any way.
~mnl e 56 Preparation of [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy3-phenyl] meth~none ~OCH2CH2--N~_~

~C

~S 0~ 11 H3C~ CH3 y-~ ` i i WO 95/17095 . ~ `. ! . 2 ~ 7 6 PCT/US94/14~55 In dry tetrahydrofuran (250 ml) ~6-hydroxy-2-(4-hydroxyphenyl)-benzo[b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxyphenyl]-meth~none, hydrochloride (5.1 g, 10 ~mol) was suspended and 7.1 g (70 mmol) of triethylamine was added. The reaction mixture was cooled to 0C in an ice bath and 10 mg of 4-dimethyl~mi nopyridine (DMAP~ was added, followed by the slow addition of n-butylsulfonyl chloride (4.7 g, 30 mmol). The reaction mixture was placed under a nitrogen atmosphere and allowed to warm slowly to room temperature and continued for 72 hours. The reaction mixture was filtered and evaporated to an oil. The oily residue was dissolved in chloroform and chromatographed on a silica gel column and eluted wi~h a linear gradient of chloroform to chloroform-methanol (19:1; V:V). The desired fractions were com~bined and evaporated to dryness to afford 5.60 g of the title compound as a tan amorphous powder.

C36H43NO8s3 20 ~S (FD) m~e=714 (M+l) NMR was consistent with the proposed structure.

Fx~m~le 57 Preparation of ~6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzotb]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone, Hydrochloride ~OCH2 CH2--N~_~

~SI-o~
H3C~ HCl CH3 WO95/17095 ~ . . ; 2 1 76 ~ 27 PCT/US94/14655 The commpound of Example 1, [6-(n-Butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]-benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone (5.4 g) was dissolved in ethyl acetate (EtOAc) and a solution of ether, saturated with hydrochloric acid, was added until no more precipitate was formed. The liquid was decanted off and the solid was triturated with ether. The title compound was crystallized from hot ethyl acetate to afford 3.74 g, as a white powder.
C36H43WOgS3-HCl Elemental Analysis: C H N
Calculated:57.7 5.88 1.87 Found: 57.75 5.93 1.93 NMR was consistent with the proposed structure.

F.~mnl e 58 Preparation of [6-(n-pentylsulfonoyl)-2-[4-(n-pentylsulfonoyl)phenyl~benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone ~ocH2cH2--N~

~C

o:p8 ~ CC~O--S
~S--O

In dry tetrahydrofuran (100 ml) o~ [6-hydroxy-2-(4-hydroxyphenyl)-benzo[b]thien-3-yl]-[4-[2-~1-piperidinyl)ethoxy]phenyl-]meth~no~e, hydrochloride (3 g, 5.9 WO 95/17095 ~ . 2 t 7 6 1 2 7 PCT/US9~/14655 mmol) was suspended and 10 mg of DMAP was added followed by 3 g (30 mmol) of triethylamine. The reaction mixture was stirred at room temperature and under a nitrogen blanket for about 20 minutes. n-Pentyl sulfonyl chloride (2.5 g, 14.7 mmol) was dissolved in 25 ml of tetrahydrofuran and slowly added to the stirring reaction mixture. The reaction was allowed to proceed at room temperature and under nitrogen for eighteen hours. The reaction mixture was filtered and the volatiles were removed i~ v~cl~o. The resulting material was dissolved in a small amount of chloroform and chromatographed (HPLC) on a silica gel column eluted with a linear gradient starting with chloroform and Pn~;n~ with chloroform-methanol (19:1 v/v). The desired fractions were determined by thin layer chromatography, combined and evaporated down to afford 3.82 g of the title compound as thick oil.

C3gH47No8s3 NMR: consistent with the proposed structure MS: (FD~ m/e=743 (M+2) 20 Elemental Analysis: C H N
Calculated: 61.51 6.39 1.89 Found: 57.63 6.44 1,50 ~mn 1 e 59 Preparation of [6-(n-pentylsulfonoyl)-2-[4-(n-pentylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone, Hydrochloride WO 95/17095 `, :; 2 1 7 6 t 2 PCTIUS94/14655 ~OCH2CH2--N~

~C

~ 5l"~

HCl [6-(n-Pentylsulfonoyl)-2-[4-(n-pentylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone (3.7 g) was dissolved in 25 ml of ethyl acetate and a solution of hydrochloric acid saturated diethyl ether was added. A precipitate formed and the liquid decanted off. The gummy solid was triturated with diethyl ether and dried n v~cno at room temperature to afford 2.12 g of the title compound as a white amorphous and hygroscopic solid.

C38H47NOgS3 HCl NMR: consistent wi.th the proposed structure 15 Elemental Analysis: C H N
Calculated: S8.63 6.22 1.80 Found: 57.35 6.45 1.38 F.~mnl e 60 Preparation of [6-(n-hexylsulfonoyl)-2-[4-(n-hexylsulfonoyl~phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] meth~none WO95/17095 ,~ 2 1 7 6 1 2 7 PCT/US94/14655 ~oCH2CH2--N~

- ~ C)~

O~e_ ~\o-~

In dry tetrahydrofuran (250 ml) 3 g (5.9 mmol) of [6-hydroxy-2-(4-hydroxyphenyl)-benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]-methanone hydrochloride was suspended and 10 mg of DMAP was added. Triethylamine (4 g, 40 mmol) was then added and the reaction mixture was stirred for 20 minutes at room temperature under a nitrogen blanket.
n-Hexylsulfonyl chloride (3.6 g, 19.6 mmol) in 25 ml of tetrahydrofuran was slowly added to the reaction mixture.
The reaction was allowed to proceed at room temperature and under nitrogen for 3 days. The reaction mixture was evapora~ed down in vacno and resuspended in ethyl acetate and washed with water. The organic layer was dried by filtering it through anhydrous sodium sulfate and evaporated to a yellow oil. The oil was dissolved in chloroform and chromatographed (HPLC) on a silica gel column and eluted with a linear gradient starting with chloroform and en~; ng with chloroform-methanol (19:1 v/v). The desired fractions were determined by thin layer chromatography, comh; n e~ and evaporated down to afford 3.14 g of the title compound as a thick oil.

NMR: consistent with the proposed structure MS: (FD) m/e=771 (M+1) ~- 2~ 76~27 1~

Elemental Analysis: C H N
Calculated: 62.39 6.68 1.82 Found: 62.33 6.62 2.03 FxAmnle 61 Preparation of [6-(n-Hexylsulfonoyl)-2-[4-(n-hexylsulfonoyl)phe~yl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone, Hydrochloride r~
~ OCH, CH2--N~_~

~C
o~o_ ~

~ HCl CH3 [6-(n-Hexylsulfonoyl)-2-[4-(n-hexylsulfonoyl)phenyl]benzo[b3thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone (3 g) was dissolved in 20 ml of ethyl acetate and hydrochloric acid-saturated diethyl ether was added. No precipitate formed. The reaction mixture was evaporated to a thick oil and was triturated several times with diethyl ether and dried i~
y~Q at room temperature to afford 1.64 g of the title compound as a white amorphous and hygroscopic powder.

NMR: consistent with the proposed structure Elemental Analysis: C H N
Calculated: 59.67 6.50 1.74 Found: 59.47 6.59 1.77 C4oHslNogs3-Hcl WO 95117095 2 ~ 7 6 1 2 7 PCT/US9~/14655 , ~ 81 -~x~mnle 62 Preparation of [6-(n-Butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone, Citrate 2 g (2.8 mmol) of [6-(n-Butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxyj-phenyl] methanone was dissolved in 200 ml of acetone and 0.63 g (3 mmol) of citric acid was added. The reaction mixture remained at room temperature and under a nitrogen blanket for eighteen hours. The reaction mixture was evaporated 1~ v~cuo at 50 C. The reaction mixture was triturated several times with ether and dried at room temperature L~ vacuo to afford 2.35 g of the title compound as a white amorphous and hygroscopic powder.

Elemental Analysis: C H N
Calculated: 55.68 5.67 1.55 Found: 55.39 5.60 1.60 NMR: consistent with the proposed structure FxAmnle 63 Preparation of [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[3-(1-piperidinyl)propoxy]phenyl]methAnone o~ ~30CH2CH2CH2--N~

~SI-O~~
~ CH3 WO 9S/17095 ' ' PCT/US94/146~S

2.5 g (4.77 mmol) of [6-hydroxy- 2 - [ 4-hydroxyphenyl]benzo-[b]thien-3-yl][4-[3-(1-piperidinyl)propoxy]-phenyl]meth~none hydrochloride was dissolved in 100 ml of tetrahydrofuran, 3.9 g (39 mmol) of triethylamine and lOmg of DMAP were added.
The reaction mixture was stirred for 15 minutes at room temperature and under a nitrogen blanket. 4 g (25.5 mmol) of n-butylsulfonyl chloride in 15 ml of tetrahydrofuran was slowly added. The reaction was allowed to proceed for eighteen hours at room temperature and under nitrogen. The reaction was quenched with the addition of 25 ml methanol and volume reduced Ln vacuo. The crude product was chromatographed on a silica gel column, eluted with chloroform-methanol (19:1 v/v). The desired fractions were determined by thin layer chromatography, combined, and evaporated to a tan oil.

F~mnle 64 Preparation of [6-(n-bu~ylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl]~4-[3-(1-piperidinyl)propoxy]-phenyl] methanone, hydrochloride [6-(n-Butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]-benzo[b]thien-3-yl][4-[3-(1-piperidinyl)propoxy]-phenyl]
meth~none was dissolved in ethyl acetate-hexane and hydrogen chloride gas was bubbled in. The reaction mixture was evaporated down and chromatographed (HP~C) on a silica gel column eluted with chloroform and then with chloroform-methanol (19:1 v/v). The desired fractions were determinedby thin layer chromatography and combined and evaporated down to a tan amorphous powder to afford 2.5 g of the title compound.

NMR: consistent with the proposed structure MS: (FD) m/e=728 (M-HCl) WO95/17095 ~ S 2 ~ 76 1 27 PCI~/US94/1~655 ~ 83 ~

r lemen~al Analysis: C H N
Calculated: 58.14 6.07 1.83 Found: 57.90 6.05 1.82 C37H46NOgS3-HCl Fx~mnle 65 Preparation o~ [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]~enzo[b]thien-3-yl][4-~2-(1-pyrrolidinyl)ethoxy]phenyl~methAno~e.

1.5 g of [6-hydroxy-2-[4-hydroxyphenyl]benzo[b]thien-3-yl~[4-[2-~l-pyrrolidinyl)ethoxy]-phenyl]methanone hydrochloride (3 mmol) was suspended in 200 ml of tetrahydrofuran. 1.5 g of triethylamine (15 mmol) and 10 mg of 4-N,N-dimethylaminopyridine were added. The reaction mixture was stirred for several minutes under a nitrogen atmosphere.
1.56 g of n-butylsulfonyl chloride tlO mmol) was dissolved in 50 ml of tetrahydrofuran and slowly added to the reaction mixture over a twenty minute period. The reaction mixture was stirred for eighteen hours at room temperature and under a nitrogen atmosphere. The reaction mixture was evaporated to a gum i~ v~cllo. The crude product was suspended in 100 ml o~ ethyl acetate and washed with sodium bicarbonate solution and subsequently with water. The organic layer was dried ~y filteration through anhydrous sodium sulfate and evaporated to a yellow oil. The final product was crystallized from hot ethyl acetate-hexane to afford 410 mg of the title compound.
NMR was consistent with the proposed structure MS: m/e = 700 (M+l~ FD
Elemental Analysis: C H N
Calculated: 60.20 5.86 2.01 Found: 59,94 5.94 2.00 MW = 699 C35XD.lN08S

WO 95117095 ; ` ~ PCTIUS94/1465S

mn l e 5 6 Preparation of [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)-phenyl]benzo[b]thien-3-yl]-[4-[2-(1-pyrrolidinyl)ethoxy]-phenyl]methanone hydrochloride;

350 mg of [6-(n-Butylsulfonoyl)-2-[4-Butylsulfonoyl)-phenyl]benzo[b]thien-3-yl]-[4-[2-~1-pyrrolidinyl)ethoxy]-phenyl]methanone (0.5 mmol) was dissolved in 10 ml of ethylacetate and a saturated solution of hydrogen chloride in ether was added. No precipitate formed and the reaction mixture was evaporated to a gummy, white solid. The product was triturated with diethyl ether (2x) and filtered and dried Ln vacuo at room temperature to afford 220 mg of the title compound.

NMR: consistent with the proposed structure Elemental Analysis: C H N
Calculated 57.09 5.75 1.90;
Found: 57.27 5.91 1.86 MW = 736.37 C3sH4lNOsS3 - HCl F~mnle 67 Preparation of [6-hydroxy-2-[4-(n-butylsulfonoyl)-phenyl]benzo[b]-thien-3-yl]-[4-[2-(1-piperidinyl)-ethoxy]phenyl]methanone.

~, , ~ -WO 95117095~ ~ . 2 1 7 6 1 2 7 PCT/US94/14655 ~1 '` ~s ~ocH2cH2--N~>

HO~O--S~
~ CH3 20 g of [6-hydroxy-2-[4-hydroxyphenyl]benzo[b]thien-3-yl]-[4-t2-(1-piperidinyl)ethoxy]phenyl]methanone (Raloxifene) hydrochloride (0.04 mol) was suspended in 250 ml of tetrahydrofuran. Ten grams of triethyl~m;ne (0.1 mol~ and 10 mg of 4-N,N-dimethylAminopyridine were added. The reaction mixture was stirred for several minutes under nitrogen. 6.25 g of n-butylsulfonylchloride (0.04 mol) was dissolved in 2S
ml of tetrahydrofuran and slowly added to the reaction mixture over a period of twenty minutes. The reaction was allowed to continue for 5 days at room temperature and under nitrogen atmosphere. The reaction mixture was evaporated to a gum and suspended in ethyl acetate. The ethyl acetate mixture was washed successively with water, dilute sodium bicarbonate, and water. The ethyl acetate solution was dried by filteration through anhydrous sodium sulfate and evaporated to an amorphous solid.

The resulting solid was dissolved in 50 ml of methylene chloride and chromatographed (HPLC) on a silica gel column eluted with a l; npAr gradient of chloroform to chloroform-methanol (l9:1~(v/v). Four fractions were determined by thin layer chromatography and evaporated ~n vacuo to amorphous solids:
Fraction A: [6-(n-Butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(l-piperidinyl)ethoxy]phenyl] methanone, 5.4~ a WO 95/1709~ ; ` 2 1 7 6 1 ~ 7 PCI~/US94tl4655 Fraction B: [6-~lydroxy-2-[4-(n-butylsulfonoyl)-phenyl]benzo[b]-thien-3-yl]-[4-[2-(1-piperidinyl)ethoxv]phenylJmethanone, 2.~9 a.
Rf=0.50, silica gel, CHCl3-MeOH ~l9:1)v/v Fraction C: [6-(n-butylsulfonoyl)-2-(4-hydroxyphenyl)benzo[b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, 3.60 a Rf=0.41, silica gel, CHC13-MeOH (l9:1)v~v Frac~ion D: Raloxifene, 3.94 a All of Fraction B was dissolved in hot ethyl acetate and hexane was added and the title compound crystallized out to afford 1.89 g of the title compound.

NMR: consistent with proposed structure MS: m~e=594(M~l) FD
Elemental Analysiso C H N
Calculated:64.80 5.90 2.36 Found: 64.85 6.07 2.49 C32H3sN652 F.x~mnl e 68 Preparation of [6-hydroxy-2-[4-(n-butylsulfonoyl)-phenyl]benzo[b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy3phenyl]methanone hydrochloride.

1.7 g of [6-hydroxy-2-[4-(n-butylsulfonoyl)-phenyl]benzo[b~-thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy]phenyl]meth~no~e (2.86 mmol) was dissolved in ethyl acetate and a saturated solution of hydrogen chloride-diethyl ether was added. A
thick white precipitate formed. The li~uid was decanted off.
The r~m~in;n~ solid was triturated with diethyl ether (2x) and dried to afford 1.57 g of the title compound as a white amorphous powder.

NMR: consistent with the proposed structure.

WO 95/17095 ~ 5 2 t 7 6 ~ 2 7 PCT/US94/14655 Elemental Analysis: C H N
Calculated: 60.99 5.76 2.22;
Found: 61.17 5.88 2.27 MW = 630.23 5 C32H3sNO6S2 - HCl MS: m/e = 594 (M-HCl)F.D.

~x~mn l e 69 Preparation of [6-n-butylsulfonoyl-2-[4-hydroxyphenyl]benzo~b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy]phenyl]methanone.

~OCH2 CH2--N3 ~C

o )~~ OH

All of fraction C from Example 67 was dissolved in 50 ml of hot ethyl acetate and hexane. No crystallization occurred The solvents were evaporated i~ V~CllO to afford 3.17 g of the title compound as oily, white solid.
NMR: consistent with the proposed structure.
MS: m/e = 594 (M+1) FD
Elemental Analysis: C H N
Calculated: 64.84 5.90 2.36.
Found: 64.37 5.87 2.28.
MW = 593 C32H3sNO6S

WO 95/17095 C t~ ~ r ~ 2 1 7 6 1 2 7 PCTIUS9~/14655 F.x~mn 1 e 7 0 Preparation of [6-n-butylsulfonoyl-2-[4-hydroxyphenyl]benzo[b]thien-3-yl]-[4-[2-tl-piperidinyl)ethoxy]phenyl]methanone hydrochloride.

3 g of [6-n-butylsulfonoyl-2-[4-hydroxyphenyl]-benzo[b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy]-phenyl]meth~no~e was dissolved in 50 ml of ethyl acetate and a solution of diethyl ether saturated wi~h hydrogen chloride was added. A thick white precipitate formed and the liquid was decanted off.
The solid was triturated (2x) with diethyl ether and dried.
This afforded 2.51 g of the title compound as a white amorphous powder.
NMR: consistent with the proposed structure.
Elemental Analysis: C H N
Calculated: 60.99 5.76 2.22;
Found: 60.71 5.84 2.21 MW = 630.23 C32H3sNO6S2 - HCl MS: m/e = 594 (M-HCl) F.D.

F.~mnl e 71 Preparation of [6-[N-(4-chlorophenyl)carbamoyl]-2-[4-[N-(4-chlorophenyl)carbamoyl]phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl] meth~no~e.

~ ocH2c~2-N ~

Cl ~ ~ O~C~ N ~ Cl 95/17095 ~ 76 1 27 PCT~Sg4/1~65S

5.~6 g (10.7 mmol) of [6-hydroxy-2-(4-hydroxyphenyl)benzo~b]thien-3-yl]-!4-[2-(1-piperidinyl)ethoxy~phenyl]meth~no~e was dissolved in 200 ml of dry tetrahydrofuran and 5.45 g (35.2 mmol) of 4-chlorophenyl isocynate was added. The reaction mixture was stirred at room temperature under an atmosphere of nitrogen.
After 18 hours, the solvent was removed by evaporation V~Ct~O, and redissolved in chloroform. The chloroform solution was cooled to -20C for 24 hours and the precipitate formed was filtered off. The r~m~-nin~ solution was chromatographed (Waters Prep 500, HPLC) on a silica gel column, eluted with a linear gradient of chloroform ending with chloroform-methanol (l~:l)(v/v). The desired fractions were determined by thin layer chromatography, combined and evaporated to dryness to afford 4.01 g of the title compound as a tan amorphous powder.

C42C3sCl2N3O6s 20 Elemental Analysis: C H N
Calculated: 64.64 4.48 5.38 Found: 65.69 4.81 4.83 MS (FD) m/e=779,781 ~x~mnl e 72 Preparation pf [6-~N-(4-chlorophenyl)carbamoyl~-2-~4-[N-(4-chlorophenylJcarbamoyl]phenyl]benzotb]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl] meth~no~e hydrochloride.
4.01 g of [6-[N-(4-Chlorophenyl)carbamoyl]-2-[4-[N-(4-chlorophenyl)carbamoyl]phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl] methanone was dissolved in 200 ml of ether and a small amount of tetrahydrofuran added to affect solution. A solution of ether, which had been saturated with hydrogen chloride, was added until no further precipitate formed. The reaction mixture was evaporated to WO95/17095 . ~ 2 1 ~6 1 27 PCT/US9~/146~5 dryness and triturated with ether several times. An attempt was made to crystalize the salt from hot ethyl acetate and absolute EtOH, which did work. Evaporation of the solvent, afforded 2.58 g of the title compound as a tan amorphous powder.

C42H35cl2N3o6s-Hcl Elemental Analysis: C H N
Calculated: 61.73 4.44 5.14 Found: 57.43 4.29 4.19 NMR: Consistent with the proposed structure and contains an indeterminate amount of solvent.

F.x~mn l e 73 Preparation of [6-(N-(n-butyl)carbamoyl]-2-[4-(N-(n-butyl)carbamoyl)phenyl]benzo[b]thien-3-yl]~4-[2-(1-piperidinyl)ethoxy]phenyl] methanone.

~OCH2CH2--N~>

~C

H3C~-- 1' ~O~C~ N--~'~ CH3 4.47 g (9 mmol) of [6-hydroxy-2-(4-hydroxyphenyl)benzo-[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]meth~nonP
was dissolved in 250 ml of tetrahydrofuran and 4 g (40 mmol) of n-butylisocyanate was added. The reaction mixture, at room temperature and under nitrogen, was allowed to react for 72 hours. The reaction mixture had evaporated by the end of this time and the residue was dissolved in a m;n;m~l amount WO 95/17095 ..~ t ~ 2 ~ 7 6 ~ 2 7 PCT/US94/146~i5 of chloroform. This solution was chromatographed (HPLC) on a silica gel column, eluted with a linear gradient of chloroform to chloroform-methanol (19:1) to afford 4.87 g of the title compound as a tan amorphous powder.

Elemental Analysis: C H N
Calculated: 67.73 6.75 6.52 Found: 66.43 6.67 6.24 MS (FD) m/e=672 (M+1) NMR was consistent with the proposed structure.

~x~mnl e 74 Preparation of [6-(N-methylcar~amoyl)-2-[4-(N-methylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone.

~OCH2CH2--N~

~C

H3C~ ~ ~ C~ o~ H

A suspension of 3 g (5.9 mmol) of [6-hydroxy-2~4-hydroxyphenyl)]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]meth~no~e hydrochloride in 250 ml of anhydrous tetrahydrofuran was prepared. To this suspension was added 2 g (10 mmol) of triethylAm;ne and the reaction mixture was stirred at room temperature for a~.oximately 15 minutes under a nitrogen atmosphere. To the stirring mixture was added 5.8 g (20 mmol) of methylisocyanate. The reaction was allowed to continue for 36 hours, The reaction mixture was filtered and evapoated to dryness ;n v~cl~O. The residue was dissolved in 30 ml of WO95/17095 . ~: ! . 2 1 76 1 27 PCTtUS94/14655 chloroform and chromatographed (HPLC) on a silica gel column, eluted with a linear gradient of solvent of chloroform to chloroform-methanol (19:1). The fractions were analyzed by thin layer chromatography and the desired fractions were 5 combined and evaporated to dryness i n vacllo to afford 2.2 g of the title compound as an amorphous powder.

NMR: Consistent with the proposed structure.
IR: 3465, 2942, 1741 cm-1 (CHCl3) MS: m/e=588 (M+1) FD
C32H33N306S .

~m~le 75 Preparation of [6-~N-methylcarbamoyl)-2[4-~N-methylcarbamoyl)-phenyl]benzo[b]thien-3-yl][4-[2-~1-piperidinyl)ethoxy]phenyl]me~h~n~e Hydrochloride.

Two grams of the compound of [6-(N-Methylcarbamoyl)-2-[4-(N-methylcarbamoyl)-phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone was disolved in 20 ml of ethyl acetate and a solution of hydrochloric acid-ether was added, forming a white precipitate. The reaction mixture was evaporated to dryness ; n V~ cllo . The solids were crystallized from acetone-ethyl acetate, filtered and washed with ethyl acetate and dried to afford 1.98 g of the title compound.

NMR: Consistent with the desired structure.
Elemental Analysis: C H N
Calculated: 61.58 5.49 6.73 Found: 61.25 S.96 5.97.
C32H34clN3o6s -WO 95/17095 r ~ 7 6 ~ 2 7 PCT/US94/14655 . ~. . ! ~ ~

F.x~mn I e 76 Preparation of [6-(N-ethylcarbamoyl)-2-[4-(N-ethylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone.
.

r~
~ OCH~ CH2--N~>

~C

H3C~N~ ~~0~ ~N~cH3 4 g (7.85 mmol) of [6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]meth~nQ~e hydrochloride was suspended in 250 ml of anhydrous tetrahydrofuran and 3 g (30 mmol) of triethylamine was added. The reaction mixture was stirred at room temperature under nitrogen for 15 minutes.
1.67 g ~23.5 mmol) of ethylisocyanate was added. After 24 hours, the reaction was checked by thin layer chromatography, and was not complete. An additional 4.5 g of the isocyanate was added. After 96 hours, the reaction mixture was filtered and chromatographed as in Example 74 to afford 4.23 g of the title compound as a white amorphous powder.

NMR: Consistent with the proposed structure.
MS: m/e=616 (M+l) FD
C3~H37N306S -~x~mrle 77 Preparation of [6-(N-ethylcarbamoyl)-2-[4-(N-ethylcarbamoyl)phenyl]benzo[b]thien-3-yl]t4-[2-(1-piperidinyl)ethoxy]phenyl]meth~ no~ e hydrochloride.

WO 95117095 . 2 1 7 6 1 2 7 PCT/US94/146~i5 This compound was prepared by substantially the sameprocedures of Example 75, to afford 3.58 g of the title compound.
NMR: Consistent with the proposed structure.
Elemental Analysis: C H N
Calculated: 62.61 5.87 6.44;
Found: 62.33 6.16 6.41.
C34H38ClN306s~

FxAmnle 78 Preparation of [6-(N-iso~y~lcarbamoyl)-2[4-(N-iso-propylcarbamoyl~phenyl]benzo~b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone.
r~
~OCH2CH2--N~_~

~C

CH~ ~C~, ~ ~C~ ~CH

4 g (7.85 mmol) of [6-hydroxy-2-(4-hydroxyphenyl)-benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]-methanone hydrochloride was suspended in 250 ml of anhydrous tetrahydrofuran and 3 g (30 mmol) of triethyl~m;ne was added.
The reaction mixture was stirred for 15 minutes at room temperature and under nitrogen. 2.77 g (32.6 mmol) of isopropylisocyanate was added. After 24 hours, the reaction was checked by thin layer chromatography for completeness and was not complete. An additional 10.8 g (130.4 mmol) of the isocyanate was added and the reaction was allow to continue for another 96 hours. The desired compound was isolated WO 95/17095 ~, ~ ; 2 t 7 6 ~ 2 7 PCT/US94/14655 substantially according to the procedures described in Example 19 to afford 4.01 g of the title compound as a tan amorphous powder.

5 NMR: Consistent with the proposed structure.
MS: m/e=644 tM+1) FD
C36H~lN306S -~ x~mnl e 79 Preparation of [6-(N-iso~o~ylcarbamoyl)-2-[4-(N-isopropylcarbamoyl)phenyl]benzo[b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy]phenyl]methanone hydrochloride.

This compound was prepared by substantially following the procedures of Example 75 to afford 3.58 g of the title compound as a white crystalline powder.

NMR: Consistent with the proposed structure.
Elemental Analysis: C H N
Calculated: 63.56 6.22 6.18 Found: 63.63 6.52 5.95 C36H42ClN3O6s-~x~mn 1 e 80 Preparation of [6-(N-cyclohexylcarbamoyl)-2[4-(N-cyclohexylcarbamoyl)phenyl]benzo[b]thienyl-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone.

WO 95/1709~ ~ 2 1 7 ~ t 2 7 PCT/US94/14655 ~OCH~ CH~--N~

~C

H H ~

3 g (5.9 mmol) of [6-hydroxy-2-(4-hydroxyphenyl)-benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]-methanone hydrochloride was suspended in 250 ml of anhydroustetrahydrofuran and 2 g (20 mmol)of triethylamine was added.
The reaction mixture was stirred for 15 minutes at room temperature under nitrogen. 14.5 g (105 mmol) of cyclohexylisocyanate was added. The reaction was allowed to continue for 48 hours, then an additional 20 mmol of the isocyanate was added. After a further 24 hours, the desired product was isolated substantially according to the procedures of Example 19 to afford 4.07 g of the the title compound as a tan amorphous powder.
NMR: Consistent with the proposed structure.
MS: m/e=724 (M+1) FD

Fx~mnle 81 Preparation of 6-(N-cyclohexylcarbamoyl)-2-[4-(N-cyclohexylcarbamoyl)phenyl]benzo[b]thienyl-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]meth~nQne Hydrochloride 3.9 g of 6-(N-cyclohexylcarbamoyl)-2[4-(N-cyclohexylcarbamoyl)phenyl]benzo[b]thienyl-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone was converted to its hydrochloride salt by substantially the same procedures as described for Example 75 and crystallized from hot ethyl WO 9S/17095 2 ~ 7 6 t 2 7 PCT/US94114655 ., ,~ i .~ t _ 97 _ acetate. This afforded 3 g of the title compound as a white powder.

NMR: Consistent with the proposed structure.
5 Elemental Analysis: C H N
Calculated: 66.34 6.63 5.53 Found: 66.32 6.92 5.62 C~2H50ClN306S .

~x~mnl e 82 Preparation of [6-(N-phenylcarbamoyl)-2-~4-(N-phenylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone.
r~
~OCH2CH2--N~J

~C

~ ~C~ ~ H~

3 g (5.9 mmol~ of [6-hydroxy-[2-(4-hydroxyphenyl)benzo [b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]meth~none hydrochloride was suspended in 250 ml of anhydrous tetrahydrofuran and 2 g (20 mmol) of triethylamine was added.
The reaction mixture was stirred for 15 minutes at room temperature under nitrogen. 15 ml of phenylisocyanate was added and the reaction was allow to continue for 96 hours.
An additional 5 ml of isocyanate was added. After a further 48 hours, the reaction mixture was filtered and evaporated to an oil, The oil was triturated with heptane and the li~iud decanted off. The oil was dissolved in chloroform and chromatographed (HPLC) on a silica gel column, eluted with a linear gradient of chloroform to chloroform-methanol (19:1).

i ~

9821 7~7 The desired fractions were combined and evaporated to an oil to afford 3.31 g of the title compound.

NMR: Consistent with the proposed structure.
MS: m/e=711 and some 212 (diphenylurea) C42H37N306S .

F~Amnle 83 Preparation of [6-(N-phenylcarbamoyl)-2-[4-(N-phenylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone Hydrochloride.

3.2 g of [6-(N-phenylcarbamoyl)-2[4-(N-phenylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methAno~e was disolved in ethyl acetate and filtered. Hydrogen chloride-ether was added to the solution and a white precipitate formed. The li~uid was decanted off. The solid was dissolved in a small amount of acetone and ~iltered, then it is was evaporated to dryness to afford 270 mg of the title compound as a tan amorphous powder.

Elemental Analysis: C H N
Calculated: 67.42 5.12 5.62 Found: 67.51 5.37 5.50 C42H38clN3o6s -By substantially following the procedures described above one skilled in the art can prepare the other compounds of Formula I.

~-Amvloi~ Pro~uct;on Tnh;h; tion A~SAV
Two cell lines [human kidney cell line 293 and Chines hamster ovary cell line (CHO)] were stably transfected WO95/17095 ~ 2 1 7 5 ~ 2 7 PcT~ss4/l465~

_ 99 _ with the gene for amyloid precursor protein (APP-751) cont~i n ng the double mutation Lys6sl-Met6s2 to Asn6sl-~eu6s2 (APP-751 numbering system) commQnly called the Swedish mutation using the method described in Citron, et ~l., N~tllre (T.on~on), 360:672-674 (1992). The transfected cell lines wre designated as 293 7~1 SWE and CHO 751 SWE, and were plated in 96-well plates at an initial density of 2.5 x 10~ and 1 x 104 cells per well, respectively, in Dulbeccols ~i n i mA 1 Essential Media (DMEM) supplemented with 10% fetal bovine serum.
Following overnight incubation at 37 C in an incllhAtor equilibrated with 10% carbon dioxide, the media was removed and replaced with 200 ~l per well of DMEM cont~in;ng test compound at various concentration. Controls were also performed in which no test compound was added to the media in 1~ the wells. After a two hour pretreatment period, the media werew again removed and replaced with fresh media supplemented with the test compound and the cells were incubated for an additional two hours.
The test compound stocks were prepared in dimethylsulfoxide (DMS0) such that, at the highest conentra~ion of tet compound, the concentration of DMSO in the wells never exceeded 0.5~.
After treatmen~, plates were centrifuged at 1200 rpm for about five minutes at room temperature to pellet cellular debris from the conditioned media. From each well, 100 ~1 of conditioned media were transferred into an enzyme-linked ;mmllnosorbent assay (ELISA) plate pre-coated with antibody 266, antibody specific for ~-amyloid(13-28). D.
Seu~ert, et al., N~tllre (Ton~on~, 359:325-327 (1992). These ELISA plates were then stored at 4-C overnight. An ELISA
assay employing labeled antibody 6C6, an Antihody specific for ~-amyloid~1-16), was run the next day to measure the amount of ~-amyloid peptide produced.
The cytotoxic effects of the compounds of this invention were measured by a modification of the method of Hansen, et ~l., Jol7rn~1 of Tmml~noloa;c~l Methods, 119:203-210 (1989). To the cells rPm~ i n i ng in the tissue culture plates Ir WO95/17095 ~ - ~ 2 1 76 1 27 PCT/US9~/146!;5 after the pelleting above was added 25 ~1 of 3-(~,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ~MTT) stock solution (5 mg/ml) to a final concentration of 1 mg/ml.
The cells were incubated at 37 C for about one hour at which time the cellular activity was stopped by the addition of an equal volume of MTT lysis buffer (20% w/v sodium dodecylfulfate in 50% N,N-dimethylformamide, pH 4.7).
Complete extraction was achieved by overnight shaking at room temperature. The difference in the optical density at 562 nm (ODs62) and at 650 nm (OD6so) was measured in a spectrophotometer as an indicator of cellular viability.
The results of ~-amyloid ELISA were fit to a st~n~rd curve and expressed as ng/ml of ~-amyloid peptide.
In order to normalize for cytotoxicity, the ~-amyloid concentrations were divided by the MTT results and expressed as a percentage of the results from the drug-free controls.

Many of the compounds of Formula I were very active in the inhibition of ~-amyloid production in the above assay while not causing significant cytotoxicity. The aggregation of ~-amyloid peptide results in the activation of immune and inflammatory responses in affected areas of the brain. The ~-amyloid peptide, upon assuming the conformation necessary for aggregation, also potentiates cytokine secretion, especially interleu]~in-6 and interleukin-8 release. The compounds of Formula I, in preventing the formation of ~-amyloid aggregates, inhibit this cytokine release and, thereby, ameliorate the inflammatory component of Al7he;m~r's Disease.
The compounds of Formula I are usually ~m;ni stered in the form of pharmaceutical compositions. These compounds can be ~m; n; stered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intr~n~l. These compounds are effective as both injectable and oral compositions. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.

WO 95/17095 ~ ` 2 ~ ~ 6 ~ Z 7 PCT/US941146SS

The present invention also includes pharmaceutical compositions which contain, as the active ingredient, the compounds of Formula I associated with pharmaceutically acceptable carriers. In making the compositions of the present invention the active ingredient is usually mixed with an excipient, diluted ~y an excipient or enclosed within such a carrier which can be in the form of a capsule, cachet, paper or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments contA; n 1 ng for example up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.
In preparing a formulation, it may be necessary to mill the active compound to provide the a~pLoy~iate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it ordinarily is milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size is normally adjusted by milling to provide a substantiallY uniform distribution in the formulation, e.g.
about 40 mesh.
Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulations can additionally include:
lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and susp~n~i~g agents; preserving agents such as methyl- and propylhydroxybenzoates; sweetening agents; and flavoring agen~s. The compositions of the invention can be formulated WO95/1709S t ,' 2 1 76 1 27 PCT/US94/14655 so as to provide quick, sustained or delayed release of the active ingredient after ~min;stration to the patient by employing procedures known in the art.
The compositions are preferably formulated in a unit dosage form, each dosage containing from about 5 to about 100 mg, more usually about 10 to about 30 mg, of the active ingredient. The term "unit dosage forml' refers to physically discrete units suitable as unitary dosages dosages for human subjects and other m~m~ 1 S, each unit cont~; n; ng a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient.
For examples, dosages per day normally fall within the range of about 0.05 to about 30 mg/kg of body weight. In the treatment of adult hllm~n~, the range of about 0.1 to about 15 mg~kgJday, in single or divided dose, is especially preferred. However, it will be understood that the amount of the compound actually ~m; n; stered will be determined by a physician, in the light of the relevant circumstances, including the condi.tion to be treated, the chosen route of ~m; n; stration, the actual compound ~m; n; stered, the age, weight, and response of the individual patient, and the severity of the patient~s symptoms, and therefore the above dosage ranges are not intended to limit the scope of the invention in any way. In some instances dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several smaller doses for ~m;n; st~ation throughout the day.
For prophylactic applications, the compounds of Formula I are A~m; n; stered to a host susceptible to Alzheimer'S Disease or another physiological condition associated with an amyloidogenic peptide, but not necessarily 3~ already suffering from such disease. Such hosts may be identified ~y genetic screening and clinical analysis, as .. . J j ~_ ~ WO 95/17095 2 ~ 7 6 ~ 2 7 PCT/US94/1465~i described in the medical literature. See e.a., Goate, N~tllre 349:704-706 (1991).
For preparing solid compositions such as tablets the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition cont~;nin~ a homogeneous mixture of a compound of the present in~ention. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This solid preformulation is then subdivided into unit dosage forms of the type described above containing from 0.1 to about 500 mg of the active ingredient of the present invention.
The tablets or pills of the present invention may be coated or otherwise compounded to provide a dosage form affording the ad~àntage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into`
the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.
The liquid forms in which the novel compositions of the present invention may be incorporated for a~m;n;stration orally or by injection include aqueous solutiona, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and s;m; l~r pharmaceutical vehicles.
Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, 2~76~27 WO95/17095 PCT~Sg~/14655 _ aaueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described t su~ra. Preferably the compositions are administered by the 5 oral or nasal respiratory route for local or systemic effect.
Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be breathed directly from the nebulizing device or the nebulizing device may be attached to a face mask, lO tent, or intermittent positive pressure breathing machine.
Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices which deliver the formulation in an appropriate mAnner~

The following examples illustrate the pharmaceutical compositions of the present invention.

FormlllAt;on F~Amnle l Hard gelatin capsules contAin;ng the following ingredients are prepared:

Quantity Tnared;ent (ma/c~sule) 25 Active Ingredient 30,0 Starch 305 0 Magnesium stearate 5,0 The above ingredients are mixed and filled into hard gelatin capsules in 340 mg quantities.

WO95/17095 ~ r 2 ~ 7 6 1 2 7 PCT~Sg4/14655 Form~ tion ~.x~m~le 2 A tablet formula is prepared using the ingredients below:

Quantity Tn~re~i~nt (ma/tAhlet) Active Ingredient 25.0 lO Cellulose, microcrystalline 200.0 Colloidal silicon dioxide lO.0 Stearic acid 5.0 The components are blended and compressed to form tablets, each weighing 240 mg.

Form~ tion ~xAmnle 3 A dry powder inhaler formulation is prepared containing the following components:

Tnare~i~nt We;aht Active Ingredient 5 Lactose 95 The active mixture is mixed with the lactose and the mixture is added to a dry powder i nhA 1; ng appliance.

WO95/17095 , ~, 2 ~ 76 ~ ~7 PCT/US94/1465~

Formul~tion Fx~m~le 4 Tablets, each cont~ining 30 mg of active t ingredient, are prepared as follows:
Quantity Inare~;ent (ma/t~hlet) Active Ingredient 30.0 mg 10 Starch ~ 45.0 mg Microcrystalline cellulose 35.0 mg Polyvinylpyrrolidone 15 (as 10% solution in water) 4.0 mg Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1.0 ma Total 120 mg The active ingredient, starch and cellulose are passed through a No. 20 mesh U.S. sieve and mixed thoroughly.
The solution of po].y~inylpyrrolidone is mixed with the resultant powders, which are then passed through a 16 mesh U.S. sieve. The granules so produced are dried at 50-60C
and passed through a 16 mesh U.S. sieve. The sodium carboxymethyl starch, m~gnesium stearate, and talc, previously passed through a No. 30 mesh U.S. sieve, are then added to the granu].es which, after mixing, are compressed on a tablet m~ch;ne to yield tablets each weighing 120 mg.

WO 95/17095 ~ . 2 1 7 6 ~ ~ 7 PCT/US94/14655 Formulation ~ le S

Capsules, each cont~ining 40 mg of medicament are made as follows:

Quantity Tnare~i~nt (m~/c~nsnle!
Ac~ive Ingredient 40.0 mg 10 Starch 109.0 mg Magnesium stearate 1 0 m~

Total 150.0 mg The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 150 mg ~uantities.

Form~ tion ~ mn l e 6 Suppositories, each cont~ining 25 mg of active ~5 ingredient are made as follows:

Tnare~;~nt ~m~n nt Active Ingredient 25 mg Saturated fatty acid glycerides to 2,000 mg The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the m; n;mnm heat necessary. The mixture is then poured into a suppository mold of nominal 2.0 g capacity and allowed to cool.

WO95/17095 ~ 2 ~ 76 ~ 27 PCT~Sg4/14655 FOr~l71 ~ tion ~ mn le 7 Suspensions, each containing 50 mg of medicament per 5.0 ml dose are made as follows:
Tn~re~;~nt ~m~lnt Active Ingredient 50.0 mg Xanthan gum 4.0 mg Sodium carboxymethyl cellulose (11%) Microcrystalline cellulose (89%) 50.0 mg Sucrose 1.75 g Sodium benzoate 10.0 mg Flavor and Color q.v.

20 Purified water to 5.0 ml The medicament, sucrose and xanthan gum are blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of the microcrystalline 2S cellulose and sodium carboxymethyl cellulose in water. The sodium benzoate, flavor, and color are diluted with some of the water and added with stirring. Sufficient water is then added to produce the required volume.

WO95/17095 ~ 2 1 7 6 1 2 7 PCT~Sg4/14655 Formul~tion ~x~mn 1 e 8 Capsules, each containing l~ mg of medicament, are made as follows:

r Quantity Tnare~ient(ma/c~nsllle) Active Ingredient 15.0 mg 10 Starch 407.0 mg Magnesium steara~e3.0 ma Total 425.0 mg The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 425 mg quantities.

For~ t;on ~m~1 e 9 An intravenous formulation may be prepared as 2 5 f ollows:

Tnare~7;~nt Ol7~nt;ty Active Ingredient 2~0.0 mg Isotonic saline 1000 ml WO 95/1709S- 2 1 7 6 1 2 7 PCTtUS94/146~5 Formulation Fxam~le 10 A topical formulation may be prepared as follows:

Tnaredi~ntoll~ntitv .~ctive Ingredient 1-10 g Emulsifying Wax30 g 10 Liquid Paraffin 20 g White Soft Paraffin to 100 g The white soft paraffin is heated until molten. The li~uid paraffin and emulsifying wax are incorporated and stirred until dissolved. The active ingredient is added and stirring is continued until dispersed. The mixture is then cooled until solid.

For~~ tion ~mn 1 e 11 Sublingual or buccal tablets, each cont~;ning 10 mg of active ingredient, may be prepared as follows:
Quantity Tn~re~;ent Per T~h]et Active Ingredient 10.0 mg Glycerol 210.5 mg 30 Water 143.0 mg Sodium Citrate 4.5 mg Polyvinyl Alcohol 26.5 mg Polyvinylpyrrolidone 15 5 ma Total 410.0 mg ~ WO95/17095 ; ~ 2 ~ 7 6 1 ~ 7 PCT~S94~146~5 The glycerol, water, sodium citrate, polyvinyl alcohol, and polyvinylpyrrolidone are admixed together by continuous stirring and maintaining the temperature at about 90C. When the polymers have gone into solution, the solution is cooled to about 50-5~C and the medicament is slowly admixed. The homogenous mixture is poured into forms made of an inert material to produce a drug-cont~- n j ng diffusion matrix having a thickness of about 2-4 mm. This diffusion matrix is then cut to form individual tablets having the a~ropLiate size.

Another preferred formulation employed in the methods of the present invention employs transdermal delivery devices (l~patches~). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds of the present invention in: controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See.
e. a ., U. S. Patent 5,023,252, issued June ll, l99l, herein incorporated by reference. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.
Frequently, it will be desirable or necessary to introduce the pharmaceutical composition to the brain, either directly or indirectly. Direct techniques usually involve placement of a drug delivery catheter into the host~s ventricular system to bypass the blood-brain barrier. One such implantable delivery system, used for the transport of biological factors to specific anatomical regions of the body, is described in U.S. Patent 5,0ll,472, issued April 30, l99l, which is herein incorporated by refernce.
Indirect techni~ues, which are generally preferred, usually involve formulating the compositions to provide for drug latentiation by the conversion of hydrophilic drugs into lipid-soluble drugs or prodrugs. Latentiation is generally achieved through blocking of the hydroxy, carbonyl, sulfate, and primary amine groups present on the drug to render the WO95/17095 2 1 76 1 27 PCT/US94/14655 ~

drug more lipid soluble and ~men~hle to transportation across the blood-brain barrier. Alternatively, the delivery of hydrophilic drugs may be enhanced by intra-arterial infusion of hypertonic solutions which can transiently open the blood-brain barrier.

Claims (7)

Claims

1. The use of a compound of the formula wherein:
A i s -O-, -S(O)m-, -N(R11)-, -CH2CH2 -, or -CH-CH-;

m is 0, 1, or 2;

x is a bond or C1-C4 alkylidenyl;

R2 is a group of the formula wherein R4 and R5 are independently C1-C6 alkyl or combine to form, along with the nitrogen to which they are attached, a heterocyclic ring selected from the group consisting of hexamethyleneiminyl, piperazino, heptamethyleneiminyl, 4-methylpiperidinyl, imidazolinyl, piperidinyl, pyrrolidinyl, or morpholinyl;

R is hydroxy, halo, hydrogen, C3-C8 cycloalkyl, C2-C7 alkanoyloxy, C1-C6 alkoxy, or phenyl, said phenyl being optionally substituted with one, two, or three moieties selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, nitro, chloro, fluoro, trifluoromethyl -OSO2-(C1-C10 alkyl) or ;

R1 is hydroxy, halo, hydrogen, C3-C8 cycloalkyl, C2-C7 alkanoyloxy, C1-C6 alkoxy, or phenyl, said phenyl being optionally substituted with one, two, or three moieties selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, nitro, chloro, fluoro, trifluoromethyl -OSO2-(C1-C10 alkyl) or ;

each R3 is independently C1-C6 alkyl, C3-C8 cycloalkyl, unsubstituted or substituted phenyl where the substituent is halo, C1-C6 alkyl or C1-C6 alkoxy;

with the proviso that when X is a bond and A is -S-, R and R1 are not both selected from the group consisting of hydroxy, methoxy, and C2-C7 alkanoyloxy;

or a pharmaceutically acceptable salt or solvate, in the preparation of a medicament useful for the treatment or prevention of a physiological disorder associated with an amyloidogenic peptide.

2. The use of a compound as claimed in Claim 1 wherein the compound is selected from the group consisting of 3-(4-methoxyphenyl)-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl-1,2-dihydronaphthalene, 3-phenyl-4-[4-(2-pyrrolidin-1-ylethoxy)benzoyl]-7-methoxy-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, 3-(4-hydroxyphenyl)-4-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-[4-[2-(hexamethyleneimin-1-yl)benzoyl]-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-7-methoxy-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-[4-[2-(N-methyl-1-pyrrolidinium)ethoxy]benzoyl]-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-[4-(2-dimethylaminoethoxy)benzoyl]-1,2-dihydronaphthalene, 3-(4-methoxyphenyl)-4-(4-diethylaminoethoxybenzoyl)-1,2-dihydronaphthalene, and 3-(4-methoxyphenyl)-4-(4-diisopropylaminoethoxybenzoyl)-1, 2-dihydronaphthalene, or a pharmaceutically acceptable salt or solvate thereof.

3. The use of a compound as claimed in Claim 1 wherein the compound is selected from the group consisting of 2-(4-hydroxyphenyl)-3-[4-[2-(hexamethyleneimin-1-yl)ethoxy]benzoyl]-6-hydroxybenzofuran, 2-(4-hydroxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-hydroxybenzofuran, 2-(4-hydroxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-6-hydroxybenzofuran, 2-(4-hydroxyphenyl)-3-[4-[2-(N,N-diethylamino)ethoxy]benzoyl]-6-hydroxybenzofuran, 2-(4-hydroxyphenyl)-3-[4-[2-(N,N-diisopropylamino)ethoxy]benzoyl]-6-hydroxybenzofuran, 2-(4-hydroxyphenyl)-3-[4-[2-(N,N-dimethylamino)ethoxy]benzoyl]-6-hydroxybenzofuran, 1-ethyl-2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-hydroxyindole, 2-(4-methoxyphenyl)-3-[4-[2-(hexamethyleneimin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran, 2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran, 2-(4-methoxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]-6-methoxybenzofuran, 2-(4-methoxyphenyl)-3-[4-[2-(N,N-diethylamino)ethoxy]benzoyl]-6-methoxybenzofuran, 2-(4-methoxyphenyl)-3-[4-[2-(N,N-diisopropylamino)ethoxy]benzoyl]-6-methoxybenzofuran, 2-(4-methoxyphenyl)-3-[4-[2-(N,N-dimethylamino)ethoxy]benzoyl]-6-methoxybenzofuran, and 1-ethyl-2-(4-methoxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]-6-methoxyindole, or a pharmaceutically acceptable salt or solvate.

4. The use of a compound as claimed in Claim 1 wherein the compound is selected from the group consisting of 2-(4-methoxyphenyl)-3-[4-[3-(hex amethyleneimin-1-yl)propoxy]benzoyl]benzo[b]thiophene, 2-(4-methoxyphenyl)-3-[4-[2-(hexamethyleneimin-1-yl)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-methoxyphenyl)-3-[4-[3-(piperidin-1-yl)propoxy]benzoyl]benzo[b]thiophene, 2-(4-methoxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-methoxyphenyl)-3-[4-[2-(N,N-diethylamino)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-methoxyphenyl)-3-[4-[2-(N,N-diisopropylamino)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-methoxyphenyl)-3-[4-[2-(N,N-dimethylamino)ethoxy]benzoyl]-benzo[b]thiophene, 2-(4-chlorophenyl)-3-[4-[2-(hexamethyleneimin-1-yl)ethoxy]benzoyl]-6-hydroxybenzo[b]thiophene, 2-(4-hydroxyphenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-hydroxyphenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-hydroxyphenyl)-3-[4-[2-(N,N-diethylamino)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-hydroxyphenyl)-3-[4-[2-(N,N-diisopropylamino)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-hydroxyphenyl)-3-[4-[2-(N,N-dimethylamino)ethoxy]benzoyl]benzo[b]thiophene, 2-(4-chlorophenyl)-3-[4-[2-(pyrrolidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene-1-oxide, and 2-(4-chlorophenyl)-3-[4-[2-(piperidin-1-yl)ethoxy]benzoyl]benzo[b]thiophene-1-oxide, or a pharmaceutically acceptable salt.

5. The use of a compound as claimed in Claim 1 wherein the compound is selected from the group consisting of [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl]methanone, [6-(n-pentylsulfonoyl)-2-[4-(n-pentylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl]methanone, [6-(n-hexylsulfonoyl)-2-[4-(n-hexylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]-phenyl]methanone, [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl][4-[3-(1-piperidinyl)propyloxy]phenyl]methanone, [6-(n-butylsulfonoyl)-2-[4-(n-butylsulfonoyl)phenyl]benzo[b]thien-3-yl]-[4-[2-(1-pyrrolidinyl)ethoxy]-phenyl]methanone, [6-hydroxy-2-[4-(n-butylsulfonoyl)-phenyl]benzo[b]-thien-3-yl]-[4-[2-(1-piperidinyl)-ethoxy]phenyl]methanone, [6-n-butylsulfonyl-2-[4-hydroxyphenyl]benzo[b]thien-3-yl]-[4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, [6-[N-(4-chlorophenyl)carbamoyl]-2-[4-[N-(4-chlorophenyl)carbamoyl]phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, [6-(N-(n-butyl)carbamoyl]-2-[4-[N-(n-butyl)carbamoyl]phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, [6-(N-methylcarbamoyl)-2-[4-(N-methylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, [6-(N-ethylcarbamoyl)-2-4-(N-ethylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, [6-(N-isopropylcarbamoyl)-2-[4-(N-iso-propylcarbamoyl)phenyl]benzo[b]thien-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, and [6-(N-cyclohexylcarbamoyl)-2-[4-(N-cyclohexylcarbamoyl)phenyl]benzo[b]thienyl-3-yl][4-[2-(1-piperidinyl)ethoxy]phenyl]methanone, or a pharmaceutically acceptable salt or solvate thereof.

6. The use of a compound as claimed in Claim 1 wherein said physiological disorder associated with an amyloidogenic peptide is Alzheimer's Disease or Down Syndrome.

7. A pharmaceutical formulation useful for the treatment or prevention of a physiological disorder associated with an amyloidogenic peptide, said formulation comprising as an active ingredient a compound as specified in any one of claims 1 to 5, associated with one or more pharmaceutically acceptable carriers, diluents, or excipients therefor.