CA2169750A1 - Di- and tetra-hydrobenzo[f]quinolin-3-ones - Google Patents

Abstract

Invented are di- and tetra-hydrobenzo¢f!quinolin-3-one compounds, pharmaceutical compositions containing these compounds, and methods for using these compounds to inhibit steroid 5-.alpha.-reductase. Also invented are processes used in preparing these compounds.

Description

WO95/05821 21 B 9 7 5~ PCTtUS91~'~5382 DI- AND TETRA- HYDROBENZO[F]QUINOLIN-3-ONES

FIF.T .n OF I~EINVF.~rllO N
The present invention relates to certain novel di and tetra-hydrobenzo[f~quniolin-3-one compounds, ph~m~e ~ti~l compositions cont~ining these co~ ou.~ds, and m~tho~l~ for using these co-l-~ounds to inhibit steroid 5-a-reduct~e Also invented are novel processes useful in pl~p~ulg these con~ounds.

nF.. ~Cp~TlYIlON OF ~F~ ~TF.l ) ART
The class of steroidal ho. .~ones known as androgens is responsible for the physical ch~nc~ tir~s that dirrt;l ,nliate males from females. Of the several organs that produce androgens, the testes produce these ho"l.ones in the greatest ~
Centers in the brain exert pl;lll~ y control over the level of androgen production.
15 NUI11~LUUS physical manifest~tion~ and disease states result when inerr~cli~e control results in excessive androgen hc)rmone production. For example, acne vulgaris, sebo,ll,ea, female hirs~lti~m, male pattern b~l-1ness and ~,u~le ~ e~ces such asbenign prostatic 1IY~ I~Y are correlated with elevated androgen levels.
ition~lly, the re(l-~ctinn of androgen levels has been shown to have a lh~dl)eulic 20 effect on prostate cancer.
Te;,lu~L~,lone is the plinci~al androgen secreted by the testes and is the ~filll~u ~ androgenic steroid in the plasma of males. It now is known that 5-a-reduced androgens are the active hol...olles in some tissues such as the ~lu~late and seb~<eous gland. Circ~ ting les~ one thus serves as a ~,ohc",llolle for 25 dihydlole~lo~lel~nc (DHT), its 5-oc-reduced analogue, in these tissues but not in others such as muscle and testes. Steroid 5-oc-reductase is a nicotin~mi~le adenine dinucleotide ~hosphale (NADPH) dependent el-zy-llc that converts lei,lo~ unc to DHI'. The i.l.~ ce of this el.zyll~c in male development was dr~m~tic~lly und~scol~,d by the disco~ely of a genetic steroid 5-a-re~luct~e deficiency in male 30 pse~ldoherm~rhrodites. Imperato-McGinley, J., et al., (1979), J. Steroid Biochem.
1 1 :637-648.

Recognition of the illl~lUl ~ulce of elevated DHT levels in various disease states has stim-ll~te~l many efforts to synthç~i7ç inhibitors of this enzyme.

A number of 5-a-redllct~e inhibiting compounds are known in the art. For example, 2,16 975~ PCT/USS1~93&2 1. J. Steroid Biochem., Vol. 34, Nos. 1-6 pp. 571-575(1989), by M.A.
Levy, et al., describes the interaction mechanism bcl~een rat prostatic steroid 5-alpha re~ ct~ce and 3-chlbuAy-17~-s~1bstituted steroids;
2. J. Med. Chem. (1990) Vol. 33, pp. 937-942, by D.A. Holt, et al., 5 describes the new steroid class of A ring aryl carboxylic acids;

3. T~S (December 1989) Vol. 10, pp. 491-495, by B.W. Metcalf, et al., describes the effect of inhibitors of steroid Sa-reductase in benign prostatic hyperplasia, male pattern b~l~lnrss and acne; and 4. EPO Publn. No. 0 343 954 A3, to D. A. Holt, et al., (SmithRline 10 Beckmann) des~ibes steroidal 3-carboxylic acid derivatives as useful 5-a-re~lnrt~e inhibitnrs.

5. EPO Publn. No. 0531026 Al, to K.S. Hirsch, et al., describes hexahydrobenzotf~quinolin-3-ones as 5-a-re~l-lr-t~ce inhi-hitors.

However, none of the above references specifir~lly suggests that any of the novel di and tetra-hyd~ zo[f~quinolin-3-one cc,lllpou"ds of the present invention would have utility as potent l~ ,s~ e 5-a-reductase inhihitors.

SUMl~AT~Y OF THE INVl~ ON
The present invention resides in the discovery that steroid 5-a-re~uct~ce is inhihite~l by certain di and tetra-hydrobenzo[f~quinolin-3-one coml)oLIl-ds. Theco~ ou~lds are potent ell~ymc inhihitors.
~selltly ~l~re.l~d comyounds of the invention and co~ oullds used in the ,nted pharm~re~ltir~l co-ll~o~;lion~ and the invented methods include:
8-chloro-4-methyl-3,4-dihydrohenzo[f~quinolin-3-one, 8-chloro- 1 '~ ,3,4-tetrahy~llul~n~o[f~quinolin-3-one, 8-methoxy-4-methyl- 1,2,3,4-tetrahydrobenzo[f~quinolin-3-one, 4-methyl-3,4-dihydrohenzo[flquinolin-3-one, 8-chloro-4-methyl-1,2,3,4-tetrahydrobenzo[f~quinolin-3-one and 8-chloro-4-methyl-3,4,5,~tetrahydrobenzo[f~quinolin-3-one.
The invention also is a m~th~l for inhibi~ing 5-a-reductase activity in ",;.."",~l~, incl~ ing hnm~n~, that comrri~es ~rlmini~tering to a subject an effective amount of a ph~rm~reutir~lly active compound of the invention. In a further aspect of the invention there are provided novel processes useful in pl~a ing the presently 35 invented 5-a-re~ ct~e inhibiting compounds. Tncl~l(le l in the present invention are pha~ cc;~l~;r~l compositions compri~ing a ph~rm~re~1tic~l carrier and compounds useful in the mPthods of the invention. Also included in the present invention are Wo 95J05821 21 6 g is~ PCT/US~11J'1~&2 mtothorl~ of co-fl~lmini~tering the pharm~eutic~lly active co~ x>ul.ds of the invention with further active ingre~lientc nF.TA~ .Fn DF.~CRl~TlON OF THF. INVF.~TION
The ~l~sellLly invented colllpoullds that inhibit 5-a-re~luct~ce have the following Formula (I):
R

R ~=
in which the A and B rings have optional double bonds where inrlic~te~l by the broken lines, provided that the A ring or the B ring or both the A and B rings have a double bond where inrlic~t~ by the broken lines;
Rl is hydl`~gel~ or -CH3; and R~ is h~log~n, or lll.,~ y;
and ph~ elltir~lly acceptable salts, hydrates and solvates thereof.

~rell~d among the ~lGse.~lly invented colllpc,ullds are those having the following formula (Ia) Rl in which the A and B rings have optional double bonds where in~lic~te~ by the broken llines, provied that the A ring or the B ring or both the A and B rings have a double bond where intli~ted by the broken lines;
R1 is hydrogen or -CH3; and R2 is halogen or meLllo,~y;
and ph~ ce~lti~lly acceptable salts, 1IY~ L~,S and solvates thereof.
Particularly pl~rcllGd among Formula (Ia) compounds are those in which R2 is chloro or methoxy, preferably chloro.

P~,Çell~d among the plesellLly invented Formula I compounds are 8-chloro-4-methyl-3,4-dihydrobenzo[flquinolin-3-one, 8-chloro-1,2,3,4-tetrahydrobenzo[flquinolin-3-one, 8-methoxy-4-methyl- 1 ,2,3,4-tetrahydrobenzo[flquinolin-3-one 8-chloro-4-methyl-1,2,3,4-tetrahy~l,obellzo[flquinolin-3-one and Wo 95/05821 2 ~ ~ 9 7 5 ~ Pcrruss4/09382 ~

8-chloro-4-methyl-3 ,4,5,6-tetrahydrohenzo[f~quinolin-3-one and ph~rrn~c~e ltic~lly acceptable salts, hydrates and solvates thereof.

S Co.l.pc,ullds of Formula (II) are included in the ~h~ "~ 1 compositions of the invention and used in the m~thol~ of the invention.

3 /~\

~
in which:
the A and B rings have optional double bonds where in~ t~ by the broken 10 lines, provided that the A ring or the B ring or both the A and B rings have a double bond where inflir~te l by the broken lines;
Rl is hydrogen or-CH3; and R3 is hydrogen, methoxy or halogen;
and ph~ .-l;c~lly flçcept~hle salts, hydl~les and solvates Ih~.~,or, 15 except co-l.~ot,nds in which Rl and R3 are both hydrogen.
,r~ d among the presently invented Formula II co~ ounds are:
8-chloro-4-methyl-3,4~ihydrobenzo[f]quinolin-3-one, 8-chloro- 1,2,3,4-tetrallyL~,~n~o[f~quinolin-3-one, 8-methoxy-4-methyl-1,2,3,4-tetrahyd~benzo[f~quinolin-3-one 20 ~methyl-3,4-dihydrobenzo[f]quinolin-3-one, 8-chloro-4-methyl-1,2,3,4-tetrahydl~,~nzo[f~quinolin-3-one and 8-chloro-4-methyl-3,4,5,~tetrallyd,uben,o[f~quinolin-3-one and ph~ ce~tic~lly acceptable salts, llyd~~Les and solvates thereof.
The term "a-receplor antagonist", as used herein, refers to a known class of 25 alpha-and~ ic n,cel)Lor antagonist col--o~ , such as described in Lafferty, et al. U.S. Patent No. 4,963,547, which are utilized in treating vascular disorders such as ~ hetes~ cardiovascular (li~e~e, benign prostatic hy~ uphy and ocular lJ~l t -~ic)n.
~Ç~llcd alpha-andrtnel ~,ic l._ce~t(~- ~nt~)ni~t~ for use in the compositions 30 and methocls of the invention include pm~lllo~in, ~ ~osin, dox~7osin, alruzo~in, in~or~min, pra7~sin~ 7-chloro-2-ethyl-3,4,5,6-tetrahydro-4-methylthieno[4,3,2-ef~[3]-bPn7~7~oFine and 8-{3-[4-(2-methoxyphenyl)-1-pipt;.~zinyl)-propylc~l,a.--oyl} -3-methyl-4-oxo-2-phenyl-4H-1-bell~ol,yldll.

WO95/05821 $'~ 75~ PCT/US94/09382 By the term "~m~nlosin" as used herein is meant a cc,~ tJulld of the u~;lu~e 'I CH3~CH2-GNHCH2CH2G~

and salts, hydrates and solvates thereof.
S ~hPmirAlly, Am~nlQ~in is-lP~i nAtyl as (-)-(R)-5-[2-[[2-(O-e~ho~yl)h~noxy)ethyl]amino]propyl]-2-methoxyben7~qnesulfor~mi~1p~
~mS111OS;n iS rlisclQse~ in U.S. Patent Number 4,703,063 and claimed in U.S. Patent Number 4,987,125 as being useful in treating lower urinary tract dysfunction.
By the term "t~ 7os;l-" as used herein is meant a coll~.,ulld of the structure il ~N' CH30~N ~N~J
I
CH30~ ~ ~ N

and salts, h~ al~s and solvates thereof.
C'h-o.mic~lly, t~aL~s~n iS rlPSip~n~t~PA as 1-(4-amino-6,7--limethoxy-2 7~1inyl)-4-[(tetrahydro 2-furoyl)call,onyl]~ e~,.,;.le Tera_osin is ~ close~l in U.S. Patent Number 4,251,532.
By the term ~1OY~O:~h- as used herein is meant a compound of the structure /--N-C0~0~, H3CO~N ~N~J ~ ,~

H3C0~ N

and salts, hydlatcs and solvates thereo ~hPmi~lly "~loy~7~sin" is tlesi~n~tPA as 1-(4-amino-6,7--limPthoxy-2-~ ,olinyl)-4-t(2,3-dihydro-1,4-ben~1ioxin-2-yl)carbonyl]-piper~7in-o.
Dc"~azo~ is disclosed in U.S. Patent Number 4,188,390.
By the term "alru~osill" as used herein is meant a compound of the structure WO 95/05821 21~ 9 7 5 a PCT/US91~'0~82 H3C0~6~N I~

and salts, hydrates and solvates thereof.
Ch~mi~lly ~lr,.,osin is ~lesign~t~ as N-[3-[(4-amino-6,7-dill,cll.oxy-2-~l...n~7~1inyl)methylamino]propyl]tetrahydro-2-furanc~L~o~ ...;r1e Alru~sin is ~1isclose~1 in U.S. Patent Number 4,315,007.
By the term ~in~-.. ;.. ~ as used herein is meant a c~ ou-ld of the ~llu~lul~, ~ ~ CH2CH2--N~ ) NHC~
and salts, hydrates and solvates thereof.
~h~.mi~lly ind~l in as design~ted N-[[1-[2-(lH-indol-3-yl)ethyl]-4-pi~ri(linyl]be~ e Tn~lo. ~- - .h~ is disclosed in U.S. Patent Number 3,527,761.
By the term "~zosin" as used herein is meant a compound of the s~ucture ~N~3 CH30~N ~N~J
CH30~ N

15 and salts, hydrates and solvates thereof.
~ hemi~lly lJIazO~ is r~ te~ as 1-(4-amino-6,7-~limetho~y-2-uina~olinyl)-4-(2-furanylc~bonyl)piper~7ine Prazosin is disclosed in U.S. Patent Number 3,511,836.
"7-chloro-2-ethyl-3,4,5,6-tetrahydro-4-methylthienot4,3,2-ef~ -20 [3]ben7~7~,pine" as used herein is meant a co-ll~oulld of the ~llu~;lulc;
C~l N-CH3 (/ \~

and salts, hydrates and solvates thereo WO 95/05821 5 PCT/US91~-~9382 7-chloro-2-ethyl-3,4,5,6-tetrahydro-4-methylthieno[4,3,2-ef~-[3]ben7~7epine is disclosed in U.S. Patent No. 5,006,521. ~ lition~lly, all cc ~ ou-lds disclosed in U.S. Patent No. 5,006,521 as alpha-acLe.~clgic receptor antagonist are preferredalpha-a~n~gic l~c~lor antagonist as used herein.

"8-{3-[4-(2-methc,~yl,hc..yl)-1-pip~azinyl)-propylc~l,~l~oyl}-3-methyl-4-oxo-2-phenyl-4H-1-l~nzopyl~l" as used herein is meant a compound of the ~I-ul;lul~

N N --~

and salts, hydrates and solvates thereof.

8- { 3-[4-(2-~ ho~cy~henyl)- l -~ dLL~yl] -propylc~u bdll~yl } -3-methyl-4-oxo-2-phenyl-4H-1-l~rlzul~yl~ul is disclosed in EPO Publn. No. 0558245 A1, to Leonal~li, et al., (Recol~ti S.A.).
~ ddition~lly all cc,lll~c ullds disclosed in EPO Publn. No. 0558245 A1, as alpha-adl~,ncr~,ic l~,c~lol antagonists are ~l~,fc lcd alpha-aL~.~el~ic l~,cc~ r15 antagonists as used herein.

Persons skilled in the art can readily ~lch~ r if a cc,lllpuulld other than one specific~lly referred to herein is a alpha-andl~n~,-gic ~ec~lu ~nt~oni~t by ntili7ing the assay described in Lafferty I. Thus, all such co-ll~Jou-lds are inr~
20 within the scope of the term "alpha-an~llcncl~ic receptor ~nt~goni~tll as used herein.

By the term "minoxidil" as used herein is meant the coll-~)ound of the structure:
O
H2N~N ~NH2 N~

Wo 95/05821 2 ~ ~ 9 ~ ~ Q PCrlUSs4/09382 ~

chemir~lly minoxidil is design~t~A as 2,4-pyrimiAine~Ai~mine, 6-(1-piperidinyl)-,3-oxide. ~inQ~iAil is the active ingredient in Rogaine~ which is sold as topical solution for stimnl~ting hair growth by the Upjohn Company, T~ IA7 l~irhig~n S The term "h~u-nAI~e inhihitor", as used herein, refers to a known class of cc,~ ounds, steroidal and non-steroidal, which prevent the conversion of androgens to estrogens, such as described in Gormley et al. Tnt.orn~tional Publication Number WO 9V18132. Arom~t~ce inhihit~r~ are disclosed in Gormley et al. as having utility in treating benign prostatic hyperplasia when used in combination with a 5-a-reAl~rt~ce inhihitor.
A ~ rel~d a~ AI~e inhibit~r for use in the cc ll-po~i~ions and methoA~ of the invention 4-(5,6,7,8-tetrahy~, .i . . .iA~7Q-[ 1 ,5-a]pyridin-5-yl)benzonillile (fadrazole). Fadrazole is Ai~rloseA in U.S. Patent No. 4,728,645. AdAition~lly, all co.,.~uunds di~closeA in Go~mley, et al. Intern~tion~l P~lhliration No. WO
9V18132 as having ~ul~AlAce inhihhing activity are plcrcll~d ~rul--~ e inhibitors as used herein.
As used herein, when a 5-a-reA~lct~e inhibitrlr, as described herein and a further active in~,-cdient or ingredients are utilized together, said 5-a-reA~lct~e inhibitor can he co-~Amini~t~red with said further active ingredient or ingredients.
By the term "co-aAmini~t~qring" and derivatives thereof as used herein is meant either ~imnll~ eous aAmini~tr~tion or any manner of sc~ale se~uell~ial arlmini~tr~tion of a s-a-reduct~ce inhibiting cc,ll-~uund, as described herein, and a further active ingredient or ingreAiPnt~, such as other co~ ounds known to treat the disease states of acne vulgaris, sebo"llea, female hirsutism, male pattern b~lAness, benign ~,~ s~te hy~t~ ~hy or prostatic adenocarcinoma or compounds known to have utility when used in cQmhin~tion with 5-a-re-i~lct~e inhibitors. Preferably, if the atlmini~tration is not simlllt~neous, the coml)uu-lds are ~Amini.~tered in a close time ~,r~ ity to each other. Furthell"u,c, it does not matter if the compounds are ~Amini~t~ red in the same dosage form, e.g. one cc)lll~ûulld may be ~Amini~t~redtopically and another com~ou-ld may be aAmini~tered orally.
As used above and throughout the rem~inA~r of the application and claims the carbons of the benzo[flquinolin~ nucleus are numbered as follows WO95/05821 ~1~97SD PCT/US94/09382 .. ~0 The novel co"l~ou"ds of Formula I of the present invention can be ~ d by mP.thorl~ outlin~A in Schemes I - V below and in the ex~mrles from the st~rting S material de-s~ribeA. in each l~ ~ecli~e scheme.

S(~F.~F. I

Me DDQ Me R2~N PhMe R ~N
~0 ~0 (a) (b) Scheme I shows ~"ll,e ,is of dihy~u~nzo[f~quinolin-3-one col"pounds of Formula (I) wh~ ;n the B ring is aromatic and in which R l is methyl. The formula (a) st~ing m~tto.ri~l~ are known and can be synth~si7Yl from available m~teri~l~using known methorl~ such as tlescribeA in EPO Publn. No. 0531026Al. Formula (b) com~ou"ds are ~ d by treating formula (a) co",l,ou"ds in a suitable 15 organic solvent, such as, toluene with an o~ci~l~nt, such as 2,3-dichloro-5,6-dicyano-1,4-l~n~uh~one and heated preferably at reflux.

SCHF.l~F. Il H

~2~= PhMe ~=

(c) (d) ~0 Scheme II shows synthesis of tetrahy&u~"zû[f~quinolin-3-one colll~oullds of Formula (I) wherein the B ring is aromatic and Rl is hydrogen. The formula (c) starting materials are known and can be synthesi7.~1 from available m~teri~l.c using known methotl~ such as described in EPO Pub!n. No. 0531026A1. Formula (d) Wo 9S/05821 2 1 6 9 7 ~ ~ PCT/US9~J~g382 ~

compounds are prepared by treating formula (c) co~ ,unds in a suit~ble organic solvent, such as, toluene, with an oxi-l~nt, such as 2,3-dichloro-5,6-dicyano-1,4-benzo~luillone, and heated, preferably at reflux.
5SC~HF.l~F. n~

Me R2~= NaH, MeI 2~=

(d) (e) Scheme m shows ~y~lLhe~is of tetrahydrobenzo[f~quinolin-3-one compounds of 10 Pormula (I), wherein the B ring is aromatic and Rl is methyl. The starting m~teri~l~ are formula (d) com~ounds from Scheme II. Formula (e) cclll~oullds arepl~cd by first treating formula (d) cc,lllpoullds in a suitable organic solvent, such as 1,2-dimethoxy ethane, with a base, such as sodium hydride, and heated, preferably at reflux. The~carl~r, the Illixlulc is treated with a methylating agent, 15 preferably methyl iodide, and heated, preferably at reflux.

S~F.MF. IV
Me Me R2~ 1 KN (S 'Me ) (a) (f) Scheme IV shows synthesis of tetrahydrobenzo[flquinolin-3-one compounds of 20 Formula I, wherein there is a Cl-C2 double bond and R1 is methyl. The starting m~t~ are form~ (a) compounds from Scheme I. Formula (f) compounds are p~ d by first treating a formula (a) colllpc,und in a suitable organic solvent suc as tetrahyd~orul~n, with a base such as pOIi.c~ ..l bis(trimethylislyl) amide, at reduced l~ . . .pe - iltllres, preferably -78C, followed by the addition of a selenation 25 agent, preferably phenyl selenyl chloride and allowed to return to room L~ lule. Tht;,tarLer, Ll~n-~ with and oxifli~ing reagent, such as hydrogen peroxide, yields formula (f) cc Ill~uullds.

wo 95/05821 16~ 75~ PCT/US91~'~3382 Schem~ V
5= DDQ,PhMe 2 (a) (e) Scheme V shows ~ylllhesis of tetrahydl~nzû[f~quinolin-3-one cc,~ oul~ds of S Formula (I) wherein the B ring is aromatic and R1 is methyl. The starting materials are foImula (a) colll~uullds from Scheme I. Porrnula (e) cc -llpounds are ~ ,d by treating formula (a) com~c,unds in a suitable organic solvent, such as, tol~lene, with an oxidant, such as 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, and heated.
Com~ounds of Formula II, in which R l is methyl and R3 is hydrogen, ar 10 ~l~ared as described in Bunting, et al. J. Or~. Chem. 1986, 51, 2068-2071.
rh;.. ~. e~";r~lly ~rcept~hle salts, hydrates and solvates of Formula (II) co,.l~ounds are formed, where ~pl~liate, by m-otho~s well known to those of skill in the art.

A l,le~.l.,d ~ cess for ~ ~hlg a colll~ound of the formula in which the A ring has an optional double bond where inrlir~ted by, the broken line;
R1 is hydrogen or-CH3; and R2 is halogen or ~elho~y;
and ~h~ ., .~c~,l ;r?lly ~cept~hle salts, hydl~les, solvates and esters thereof comprises reacting a cc,-llpc)und of the formula ~ /
R2~l~e in which Rl and R2 are as desçrihed above with 2~6~7~0 2,3-dichloro-5,6-dicyano-1,4-bellzoruillolle (DDQ)in a solvent, preferably toluene, preferably at a ~ JC~IU1G above room te.~ dture and thereafter optionally forming a ph~~ c-~l;c~lly acceptable salt, hydrate or solvate thereof.

Rec~ e the ph~ e~ lly active compounds of the invention inhibit steroid 5-a-reductase activity, they have therapeutic utility in treating rii~e~es and conditions wherein de~ ,ases in DHT activity produces the desired therarelltic effect. Such rli~e~es and con~lition~ include acne vulgaris, seborrhea, female hirsutism, male pattern b~kln~ss, prostate fii~e~es such as benign prostatic hylJGllluphy, and prostatic adenocal~ ol.l~
In ~e~ ;ng potency in inhibiting the human Sa--reclnct~ce ~,n2ylllc, the following procedure was employed:

~c~ ion of rn~mbrane p~rticulates used ~ source for recombinant steroid Sa-reduct~ isozyme 1.
Chinese h~ r~ ovary (CHO) cells conl;.;l.;.-g e~p-~,ssed, l~coll-binallt human steroid 5a-re~llct~e isoenzyme 1 (Andersson, S., Rerm~n, D.M., Jenkins, E.P., and Russell, D.W. (1991) Nature 354 159-161) were homogenized in 20 mM
pot~ m phosphate, pH 6.5, buffer con~ g 0.33 M sucrose, 1 mM
dithiothreitol, and S0 pM NADPH (buffer A) using a Dounce glass-to-glass hand homogenizer (Kontes Glass Co., Vineland, N.J.). Membrane particulates were i~ol~ted by ce-,l,irugation (100,000 x g at 4C for 60 minlltes) and resuspended in 20 mM pot~ m phosphate, pH 6.5, cont~ining 20% glycerol, 1 mM
dithiothreitol, and S0 ,uM NADPH (buffer B). The suspended particulate solution was stored at -80C.

Wo95/OS821 1 G9 ~D PCT/US91~'~93&2 ualion of ...~....k~ e p~ticulates used as source forreconlbil~al-t steroid S-a-reduc~ . iso~y",e ?
Chinese h ~ t~ ovary (CHO) cells cont~ining ex~l~ssed, recomhin~nt human steroid S-a-re~uct~ce iSOZylllC 2 were homog~ni~ç~ in 20 mM pot~sillm S phosl)h~to, pH 6.5, buffer co~ g 0.33 M sucrose, lmM lithio~ ;iLoL and SO
yM NADPH (buffer A) using a Douce hand homogenizer. ~ ..hl"ne parti~ tPs co~.l;.il.;..g the recomhin~nt human enzyme were i~ol~t~l by cen~-;rugation (100,000 x g at 4C for 60 ~ es) and l~i,usl,~,nded in 20 mM pot~ lm phosph~te, pH 6.5 col-~ g 20% glycerol, lmM dithioll~ ol, and SO yM
10 NADRH (buffer B). The sucpeT~ particulate solution was stored at -80C until used.

A~y for enzy!..Ps activities ~nd inhibitors potency.
A CQl.~ t of [14C]tei,~o~crone (SO to SS mCi/mmol) in ethanol 15 and varying ~Illolul~ls of potential inhibitor in ethanol were dc~,o~i~ed in test tubes and co,-ce-.l.a~ed to d~yness in vacuo. To each tube was added buffer, 10 ,uL
(l~iCOI~-h;~ isC;lellLy~llc 1 orisoel-zyll-e 2) or 20 yL (isoellzyl,.c 2 from human e tissue) of 10 mM NADPH and an aliquot of a steroid Sa-l~~ ; ce ~ua~ion to a final volume of 0.5 mL. Assays for human steroid Sa-re~lct~e 20 isoenzyme 1 were conducted with a sample of the l~co...hil-~nt protein ~ ,sed in CHO cells in SO mM pho5~h~te buffer, pH 7.5 while assays of isoenzyllle 2 were conrl-Tcted with a ~ en~;on of human prostatic particulates and/or recombin~nt protein G~pl~,ssed in CHO cells in SO mM citrate buffer at pH S.O.
After in~ub~ting the solution at 37C for 20 or 30 minlltes the reaction was 25 quenched by the addition of 4 mT. ethyl acetate and 0.25 ,umol each of testoste~one, Sa-dihy&utcslushl~Jne~ androst~ne~liol, and androstanedione as ca~Tiers. The organic layer was removed to a second test tube and e~,a~ul~ted to dryness in a SpeedL Vac. The residue was dissolved in 40 ~"L chlorufolll~ spotted on an individual lane of a 20 x 20 cm prech~nnell~ silica gel TLC plate (Si 250F-PA, 30 Baker Chemic~l) and developed twice with ~etc-nP:chlol~fol-l- (1:9). The radiochPmi~l content in the bands of the substrate and the products was ~lct~,....;nyl with a BIOSCAN ~m~ing Sc~nner (Bioscan Inc., W~hington, D.C.).
The ~.;.,n~ of leco~ d r~liol~bel converted to product was c~ te~l, from which enzyme &.i~ivity was ~let~Prmin~l. All in~ub~tions were conducted such that 35 no more than 20% of the substrate (testosterone) was con~l-mPA
The e~p~ lly obtained data was computer fit to a linear function by plotting the l~;ipl~ocal of the ell~y~le activity (1/velocity) against the variable W0 95/os82~ 69~5 Pcr/uS~4,~93~2--inhibitor con~entration; a~palent inhibition constants (Ki app) were determined by the Dixon analysis (Dixon, M. (1953).
The value for the inhibition constant (Ki) was c~lc~ t~-d from known procedures (Levy, M. (1989), Biochemistry. 29:2815-2824).
S All of the cc -llpC unds within the scope of this invention are useful in inhibiting steroid s-a-reduct~ce in a .~ l, including hnm~nc, in need thereo CollJp~ul.ds within the scope of this invention have been tested and have shown an activity of from 0.5 Ki(nM) to 100 Ki(nM) against iso~ylllc 1 and an activity of >2000 Ki(nM) against isozyme 2. Particularly "lerellGd among the cc,lllpc,ullds of the invention and the cc,lll~unds used in the i--~ntGd ~h~ c~l cc,.llpo~ ;o~c and il,~rented methods are 8-chloro-4-methyl-3,4-dihydrobenzo[f~quinolin-3-one, 8-chloro- 1,2,3,4-tetrahydrobenzo[f~quinolin-3-one, 8-methoxy-4-methyl- 1,2,3,4-tetrahydrobenzo[f~quinolin-3-one, 4-methyl-3,4-dihydrobenzo[f~quinolin-3-one, 8-chloro-4-methyl-1,2,3,4-tetrahyd.v~nzo[f~quinolin-3-one and 8-chloro-4-methyl-3,4,5,~tetrahy&ubenzo[f~quinolin-3-one.
The ph~rm~el1*~lly active cc,lllpou..ds of the present invention are preferably incul~ulalGd into convenient dosage forms such as c~rslllPs, tablets, or inject~hle preparations. Solid or liquid ph~rm~eutif ~1 carriers are employed.
Solid carriers include, starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, m~gnesillm st~r~te, and stearic acid. Liquid carriers include syrup, peanut oil, olive oil, saline, and water. Similarly, the carrier or diluent may include any prolonged release m~tçri~l, such as glyceryl mono~lri~,.le or glyc.,lyl flicte~r~te, alone or with a wax. The arnount of solid carrier varies widely but, preferably, will be from about 25 mg to about 1 g perdosage unit. When a liquid carrier is used, the pl~a~ion will preferably be in the form of a syrup, elixir, emlllcion~ soft gelatin c~ps~lle, sterile inject~hle liquid such as an ampoule, or an aqueous or nonaqueous liquid suspension.
The ph~-~ lG~aLions are made following conventional techniques of a ph~rm~elltic~l ch~mict involving mixing, gr~nnl~ting, and co~ ~ssing, when neces~.y~ for tablet forms, or mixing, filling and dissolving the ingre;dentc, as ~,u~liate, to give the desired oral or l~enle.~l products.
Doses of the ~l~,sGnlly invented ph~rm~relltic~lly active cull.pou-lds in a pharm~reutir~l dosage unit as described above will be an effi~aeious, nontoxic ~lualltily preferably selected from the range of 0.01 - 1000 mgtkg of active compound, preferably 0.1 - 100 mg/kg. When treating a human patient in need of WO95/05821 21 6 9 7 5~ PCT/US~ 382 steroid S-a-reA~uct~cç inhibition, the selected dose is AAmini~tPred preferably from 1-6 times daily, orally or parenterally. ~efell~d forms of palcnlel~l A(lmini~trAtiQn include topically, rectally, trAn~dprmAlly~ by injection and continuoucly by infusion.
Oral dosage units for human ~lmini~tration preferably contain from 0.1 to 500 mg S of active colnrolln-l Oral AAmini~tration~ which uses lower ~1rJs~ges is pl~ d.
P~"l, r~l Arlmini~tration7 at high dosages, however, also can be used when safe and convenient for the patient.
The methotl of this invention of inhibiting steroid S-a-reductase activity in Al~, in~ ling h~ An~, comrri~es ~r1mini~tering to a subject in need of such 10 inhibition an effective stervid S-a-re~uctA~e inhibiting _lllounL of a phh.,,~A~ ;r~lly active ccl~l~ound of the present invention.
The hl~ lioll also provides for the use of a com~vund of Formula (II) in the ..~.,..r~ of a mPAi~ ~m~Pnt for use in the inhihition of steroid 5-a-reA~.ctA~e The invention also provides for a ph~rm~ellticAl comrosition for use in the 15 L~ l n~ of benign l rv~L~Le hy~Jel LIv~lly which coInrri ~eS a coll,~vulld of Formula II and a l,hh....~cellticAlly acceptable carrier.
The invention also provides for a ~h; - . . .~eutiC~l colll~o~ilion for use in the Ll~ nl of prostatic A~l~noc~n~inoma which co~nrri~es a compound of Formula II
and aph~....A~..I;r~lly acceptable calTier.
The invention also prvvides for a ~r~cess for ~ uing a pharmAceuti~Al colll~osiLion co~ ;n;.lg a phh. ,nA~euticAlly acceptable calTier or diluent and a cvlll~ound of Formula II which comrri~es bringing the co,l,~vund of Formula II
into Aecoci~tion with the ph;~ --A~eutic~lly ~cceptAble carrier or rlilnPnt No unacceptable toxirologi~Al effects are eYrected when col~oullds of the invention are Arlminictpred in accol.l~lce with the present invention.
In ~ lition~ the l~hh~ rce~lticAlly active com~uullds of the present invention can be co-~lminict~red with further active ingreAiP-ntc, such as other com~ ullds known to treat the disease states of acne vulgaris, seborrhea, female hirsutism, male pattern bAl~lnP,ss, benign prostate hy~ ~hy or prostatic adenocal.iinoma or colll~)oullds known to have utility when used in combination with S-a-re~ ctAce inhibitors. Particularly plGfellcd is the co-A~lminic~Tation of a 5-a-re~l~lctAce inhikitor, as disclosed herein, and minoxidil for use in the l~ IIeI~t of male pattern bAl-lness. Particularly ~r~ ,d is the co-A~lminict~tion of a Sa-re lllctAce inhibitor~ as disclosed herein, and a a-r~,cG~or antagonist for use in the tre~tmPnt of benign prostatic hy~lr~hy. I~GrellGd is the co-A~lminictration of a S-a-re~lllc~ce inhibitor, as disclosed herein, and an a ~ malase inhibitor for use in the l~c~ t of benign pl~ ic hy~ phy. I~Gre~lGd is the co-~lminic~Tation of a S-a-re~l~lctAce WO95/05821 2~ Pcr/uS9~ 3~2 inhibitor~ as disclosed herein, a a -receptor antagonist and an aromatase inhibitor for use in the L ~ l of benign prostatic hY~G1~hY.
Without further elaboration, it is believed that one skilled in the art can, using the prece~ling desc~ ion, utilize the present invention to its fullest extent.
S The following E~u~les are, th~lG~olG, to be consLIued as merely illustrative and not a limit~tion of the scope of the present invention in any way.

F.Y~n~le I - cu~ ,yo-~din~ to Scheln~o- I
8-chloro-4-n~thyl-3.4-dihydro-benzorflquinolin-3-one a) 8-chloro-4-methyl-3,4-dihydro-benzo[f~quinolin-3-one 2, 3-Dichlor~5, 6-dicyano-1,4-benzoquinone (DDQ) ( 550 mg, 2.4 mmol) was added to a solution of 8-chloro-4-methyl-1,2,3,4,5,6-hexahydrobenzo[f~quinolin-3-one (200 mg, 0.8 mmol) in toluene (10 ml) and was heated to reflux. After 48 hours the reaction was filtered, concentrated, and flash cLlullla~ographed (silica gel, 30% EtOAc/h~nes) to yield the title compound (85 mg, 43%). MS (DCUNH3) m/e 244[M+H]+

FY~n~ple 2 - ccllcs~ondin~ to Scheme II
8-chloro-1. '~ 3. 4-t~tr~ lrobenzomqJlinolin-3-one a) 8-chloro-1, 2, 3, 4-tetrahydrobenzo[f~quinolin-3-one 2, 3-Dichloro-5, ~di-,y~1,4-ben~uinone (DDQ) ( 55 mg, 0.24 mmol) was added to a solntion of 8-chloro-1,2,3,4,5,6-hexahydrobenzo[f~quinolin-3-one (20 mg, 0.08 mmol) in toluene (5 ml) and was heated to reflux. After 48 h the reaction was filtered, conc~-lL-~led, and flash chromatographed (silica gel, 30%EtOAc/heY~nes) to yield the dtle com~ound (7 mg, 35%). MS (DCVNH3) m/e 232 [M+Hl+

F.x~m~?le 3 - cc,l..,s~ondin~ to Scheme III
8-chloro-4-methyl-1. 2. 3. 4-tetrahyrlrobenzorflquinolin-3-one a) 8-chloro-4-methyl-1, 2, 3, 4-tetrahydrobenzo[f~quinolin-3-one Sodium hydride (2 mg, 0.06 mmol) was added to a solution of 8-chloro-1, 2, 3, 4-tetrahydrobenzo[f~quinolin-3-one (7 mg, 0.03 mmol)in 1,2--limethoxy ethane (4 ml) and was refluxed for 2 h. Methyl iodide (0.01 ml, 0.18 mmol) was added, and the reacdon was refluxed an ~ lition~l 2 h. The reaction was treated with H20 and was extr~te~ with EtOAc. The combined organic extracts were dried (MgSO4), filtered, con~e..t~aLcd, and flash chromatographed (silica gel, 30%

~ WO 95/05821 1 69 75~ PCT/US~ 382 EtOAc-hPY~ne) to yield the title compound (Smg, 68%). MS (DCVNH3) m/e 246 [M+Hl+

F.x~ml?le 4 - col~syol~din~ to Schem.o IV
8-r~hll ro-4-In~th~yl-3.4 ~6-tetralw~llo~nzorflquinolin-3-one a) 8-chlor~4-methyl-3,4,5,6-tetrahy~Lo~nzo[f~quinolin-3-one Pot~illm bis(LIil-~ell-ylsilyl)amide (0.5 M in toluene, 3.4 ml, 1.7 mmol) was added to a solution of 8-chloro-4-methyl-1,2,3,4,5,6-hexahydrobenzo[f~quinolin-3-one (210 mg, 0.85 mmol) in THF (10 ml) at -78 degrees. After 2 h phenyl selenenyl chlnrirle (390 mg, 2.04 mmol) was added and the reaction was wa~ cd toRT and stirred 2 ~lrlition~1 hours. The reaction was treated with aqueous NH4Cl and was extr~cte~ with EtOAc. The col"~ cd ~;~L~ s were dried (MgSO4), filtered, then c(.nc~;nL~ted. The residue was dissolved in THF (10 ml) and was I~U~C,l`~,d with excess solid NaHCO3, then was cooled to 0 degrees and treated with 30% H22 ( 0.29 ml, 2.5 mmol) and was stirred for lh. Then, the reaction was treated with H20 and was eAIl~cLed with EtOAc. The combined organic eA~ s were dried (MgSO4), filtered, co.~ ~l, and flash ch~ alo~l,hed (silica gel, 30% EtOAc-k.,~ es) to yield the title co,ll~)uund (14 mg,7 %). MS (DCUNH3) m/e 246 [M+H]+
F.XA~PI.F. 5 - COII~O11~ g to Scheme V
8-"~ w~y-4-methyl-l~2~3~4-tetrally~ rLo[f~quinolin-3-one a) 8-",~ Ll,o~y-4-methyl-1,2,3,4-tetrahydrobenzo[f~quinolin-3-one 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ)(110 mg, 0.5 mmol) was added to a solution of 8-,1,~ Ll,.,~y-4-methyl-1,2,3,4,5,6-hexahydrobenzo[f~quinolin-3-one (50 mg,0.22 mmol) in toluene (10 ml) and was heated to 80C. After 4 hoursthe reaction was flash ch~ aLc~ 2 l,hed (silica gel,30% EtOAc/hexanes) to yield the title co"l~wld (17 mg, 35%). MS (DCVNH3) m/e 242 [M+H}+.

Fxample 6 An oral dosage form for ~rlmini~t~ring Formula II co"loul-ds is produced by scl~,mulg~ mixing, and filling into hard gelatin c~ps~lles the ingredients in the ~r~ol~ions shown in Table 1, below.

WO95/05821 ?~G~S~ PCT/US~ 3&2--Table I
Tn.~redients Amounts 8-chloro-4-methyl-3,4- 50 mg dihydrobenzo[flquinolin-3-one m~g-.e~ - stearate S mg lactose 75 mg F.X~PT F. 7 The sucrose, calcium sulfate dihydrate and Formula (II) cc,~ ound shown S in Table II below, are mixed and gr~nnl~tr~l in the iJr~o,lions shown with a 10%
gelatin solution. The wet ~nnle are sCi~ned, dried, mixed with the starch, talcand stearic acid, scl~.led and con-~,~ ssed into a tablet.

Table II
In~ . ~die.~ls Amo-ln 8-chloro-4-methyl-3,4- 100 mg dihy~ ,Lo[f~quinolin-3-one c~lri-lm sulfate dihydrate 150 mg sucrose 20 mg starch 10 mg talc 5 mg stearic acid 3 mg F~MPT.F. 8 8-chloro-4-methyl-3,4-dilly~lnob~n~o[f~quinolin-3-one, 75 mg, is dispursed in 25 ml of normal saline to ~,epal~i an injectable p~ ion.
While the p,ef~.,.,d ell-bo ~ F ~ of the invention are ;11~ A by the 15 above, it is to be understood that the invention is not limited to the precise instructions herein rlisclosF~ and that the right to all mr)rlifir~tions comming within the scope of the following claims is ~esel ~ed.

Claims (27)

What is claimed is:

1. A compound represented by the formula:

I

in which the A and B rings have optional double bonds where indicated by the broken lines, provided that the A ring or the B ring or both the A and B rings have a double bond where indicated by the broken lines;
R1 is hydrogen or -CH3; and R2 is halogen or methoxy; and and pharmaceutically acceptable salts, hydrates and solvates thereof.

2. A compound of claim 1 that is 8-chloro-4-methyl-3,4-dihydrobenzo[f]quinolin-3-one, 8-chloro-1,2,3,4-tetrahydrobenzo[f]quinolin-3-one, 8-methoxy-4-methyl-1,2,3,4-tetrahydrobenzo[f]quinolin-3-one, 8-chloro-4-methyl-1,2,3,4-tetrahydrobenzo[f]quinolin-3-one or 8-chloro-4-methyl-3,4,5,6-tetrahydrobenzo[f]quinolin-3-one, and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

3. A compound according to claim 1 substantially as hereinbefore defined with reference to anyone of the examples.

4. A pharmaceutical composition comprising a compound of the formula:

(II) in which:
the A and B rings have optional double bonds where indicated by the broken lines, provided that the A ring or the B ring or both the A and B rings have a double bond where indicated by the broken lines;
R1 is hydrogen or -CH3; and R3 is hydrogen, methoxy or halogen;
and pharmaceutically acceptable salts, hydrates and solvates and esters thereof; - 19 -except compounds in which R1 and R3 are both hydrogen and a pharmaceutically acceptable carrier.

5. A composition of claim 4 wherein the compound is:
8-chloro-4-methyl-3,4 dihydrobenzo[f]quinolin-3-one, 8-chloro- 1,2,3,4-tetrahydrobenzo[f]quinolin-3-one, 8-methoxy-4-methyl-1,2,3,4-tetrahydrobenzo[f]quinolin-3-one, 4-methyl-3,4-dihydrobenzo[flquinolin-3-one, 8-chloro-4-methyl-1,2,3,4-tetrahydrobenzo[f]quinolin-3-one or 8-chloro-4-methyl-3,4,5,6-tetrahydrobenzo[f]quinolin-3-one.

6. A compound according to any one of claims 1 or 2 for use in therapy.

7. A compound according to any one of claims 1 or 2 in the manufacture of a medicament for use as a steroid 5-.alpha.-reductace inhibitor.

8. A compound according to any one of claims 1 or 2 in the manufacture of a medicament for use in treatment to reduce prostate size.

9. A compound according to any one of claims 1 or 2 in the manufacture of a medicament for use in treatment of prostatic adenocarcinoma.

10. A composition according to anyone of claims 4 or 5 for use in therapy.

11. A composition according to any one of claims 4 or 5 in the manufacture of a medicament for use as a steroid 5-.alpha.-reductase inhibitor.

12. A composition according to any one of claims 4 or 5 in the manufacture of a medicament for use in treatment to reduce prostatic size.

13. A composition according to any one of claims 4 or 5 in the manufacture of a medicament for use in treatment of prostatic adenocarcinoma.

14. A process for the preparation of a compound of the formula in which the A ring has an optional double bond where indicated by the broken line;
R2 is halogen or methoxy; and and pharmaceutically acceptable salts, hydrates, solvates and esters thereof which comprises reacting a compound of the formula in which R1 and R2 are as described above with 2,3-di chloro-5,6-dicyano-1,4-benzofuinone (DDQ)in a solvent, preferably toluene, preferably at a temperature above room temperature and thereafter optionally forming a pharmaceutically acceptable salt, hydrate or solvate thereof.

15. A process for preparing a pharmaceutical composition containing a pharmaceutically acceptable carrier or diluent and an effective amount of a compound of the Formula (II) as described in claim 4 and pharmaceutically acceptable salts, hydrates, solvates and esters thereof which process comprises bringing the compound of the Formula (II) into association with the pharmaceutically acceptable carrier or diluent.

16. Use of a compound according to any one of claims 1 or 2 in the manufacture of a medicament for use in inhibiting steroid 5-.alpha.-reductase

17. A method of inhibiting steroid 5-.alpha.-reductase in mammals which comprises the administration to a mammal in need of such inhibition, an effective amount of a compound according to any one of claims 1 or 2.

18. The use of a compound according to anyone of claims 1 or 2 and an alpha-receptor antagonist compound as an active therapeutic substance which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 or 2 and an alpha-receptor antagonist compound.

19. The use of a compound according to anyone of claims 1 or 2 and an alpha-receptor antagonist compound in the manufacture of medicament for use in the treatment of benign prostatic hypertrophy which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 or 2 and an alpha-receptor antagonist compound.

20. The use of a compound according to anyone of claims 1 or 2 and minoxidil as an active therapeutic substance which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 or 2 and minoxidil.

21. The use of a compound according to anyone of claims 1 or 2 and minoxidil in the manufacture of a medicament for use in the treatment of male pattern baldness which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 or 2 and minoxidil.

22. Use of a composition according to anyone of claims 4 or 5 in the manufacture of a medicament for use in inhibiting steroid 5-.alpha.-reductase.

23. A method of inhibiting steroid 5-.alpha.-reductase in mammals which comprises the administration to a mammal in need of such inhibition, an effective amount of a composition according to anyone of claims 4 or 5.

24. The use of a composition according to anyone of claims 4 or 5 and an alpha-receptor antagonist compound as an active therapeutic substance which use consist of separate sequential or simultaneous administration of a composition according to anyone of claims 4 or 5 and an alpha-receptor antagonist compound.

25. The use of a composition according to anyone of claims 4 or 5 and an alpha-receptor antagonist compound in the manufacture of medicament for use in the treatment of benign prostatic hypertrophy which use consist of separate sequential or simutaneous administration of a composition according to anyone ofclaims 4 or 5 and an alpha-receptor antagonist compound.

26. The use of a composition according to anyone of claims 4 or 5 and minoxidil as an active therapeutic substance which use consist of separate sequential or simultaneous administration of a composition according to anyone of claims 4 or 5 and minoxidil.

27. The use of a composition according to anyone of claims 4 or 5 and minoxidil in the manufacture of a medicament for use in the treatment of male pattern baldness which use consist of separate sequential or simultaneous administration of a composition according to anyone of claims 4 or 5 and minoxidil.