Synthesis. and characterization of hie.lug,,laird, and photoactivatable neuroleptics. Novel bifunctional probes for dopamine receptors V u k k Soski{7

a n d A l f r e d M~:te~Ic~.e

i~

We have s~/m~esizcd and characterized a seliv, of ;~ovc! dcm.a~,~:, of -¢..,abl>!'~cd ;M~.~ay~aaed.e-o~'~,'< -"-'-2~+. ~{+:;{~~ ,+

~d:~.,:+:~:'~h.vd:,,/4.~'.:,~]m;%k, :~:,d:~,cbk::-~,i~ic.aget co!~m,~ ~smg ~ ~adi,cn~ {}{., ; / ; }:::>;,.¢).F~:;'.,:L

4-amm,ob~tb'g:-;S-Jk.un'~,-5(4:/b~md~iwm::2..~,4,&~.5,,&. :wxah)vbo.l:l..pprid~ff4,3d,,]md~ak" :B},+) A m i x ~ r c ~f 42q mg ,.~{ ~m~i~c H I . . ; ~ d 72::~ mgz ~ N-{ N o(4,-:azjd N -by drox~.~.uccia{midc, disv~{vcd in i*~ ~{ ol dry d{meU'~yltor~,~:m~{dc. x~,.~.,sseared ovcrifi:ab!, dlc c>:cc~,s (}f ' ,a.-~q~ -a ;tas rc~mwcd t-.', ~actmm evapor;:,do.n, and p~ K ?("0~ amJ 3{~{} rng of Nal m l~){) mi f -i-mcthyl-2-pcnta~mc u a s sdrrcd at~d ~c{]u~x:d fi~:: N} ~0 th,¢ i ~ ica~{hm rp,i',.~r¢ u a s h{{crcd. ~md ai~c >.~)I~

This comf~)und was synlh.csi/*cd in analog5' ~o ~?ompc)und A3 ex~.,'pt lhat N*(4-ae~dobcnzylJbcnpcridol ~ a used as s|arli),g m a | e d a t , l | t N M R : :,?.7 ,(m, 4.!I{, :mnnalicL 7.3 (m. 41t. aromat~ck 7.i) (m. 4H. a~',.maadc}. 6 12 (d. 211"t. - N H C O N H - trom biotinl. 5.(i) (so 21L b~:nzimidazolin}t-CH---..). 4.(~1 (s,. 2tI. = N O C t t ,,-1. 4.2

clh.~no~ yielded ~5,2 g (~i; ~,p ~73-.. ~75'C,

Z2.:2. 4./.~.[:#-:2-7)'((k~t*e p:cr~si~ IM7~I{I{! f i a & phot0,Tys{ ,tl f ff! l~{~'= C;bL{~t 13ClWCCta CC!1II'i!tI[##'A~IiO~:t'},} u,m~j ;qJp~L'L #(i ml ~i N K | / E I Y l A / Tris b~f.~:ci {>c~ a~.e ~{' i{ssuc. ] b e mrt~mnl ~d ~` ' " was dc!c~wincd b> :~>~ >~d~ii~g coridi!ion'> ~ d c,crit,cd abo~c v,c:l'c c~'~l~}+c~ m eta,~.~,iand t~,, ,, } a~cac ~bscrved fo~ lhL i:i?:., ~d,~ af~piicd

Crcc.~., 19gTk 13~tyr~phcnc~ncs wcrc derivatized by Scheme ! s h ~ s the chc'mica~ ~qructt,~rc~ ~ff bk~:my[atcd comp~und~ synthesized. We d~:rivatizcd bury-

i~trodt~cing a carbc~x}t group as suggested by Scnogles ~t aI. (19~6L The carb~x3"t gr{~up ~:~fthe resulting car-

A (BUTYROPHENONES}:

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(HEXAHYDROCARBOL!NES):

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~.qth l A ' - c a r N m y ] d i i ~ i d a z o t c and . r c a a c d ~vi,q~ a~ c x . . . . . . . eess :ef c l h q c n c d i a m i n c . Finally. the obtained aminc~ we++c b i o t i n y l a l e d +.~hh the N-hydroxy~uccim-~+,o,~] t~i>Cl~ 2 ~ ~+ ~:.mm>~,{mza] ,+nem~;,+anc~imm b~wir+c~:aa~da~c nud,:i. A rat-d :Ai, +~'{:a~+P.++'~ a s s a y c++~pi~sia# Wh;+lmam, (;+::/(" +.me,r- aas t:,cd ~.g,~:~k~dc~ +d.. ~.',51, ,~cc a:t,m M,~wria~-ard .mc~i'+ud-~cc~',~ AI'I c×p~-+mscnlsv,c~c pc,-.qm~cd :~ 23(L incub;,~i+m,.~c+c p,.cHormcd b'.+

3.2, Equilil)rimn binding slu,: +i~

tac+~.pclm.et+~md:~~c>~;,, ~++cm, l:,Ol~nd the hcxahydr,:,carbolinc and phcr~othiazh"~e d,~ti'val/vJ~ A: (°~mlpcIilto~t ~iih bul~l'~phcrtollcs: ( !. i \ l t I ,; } AT; ( ~ ) AJ; { i t A & { I ) ~%+5.:( I 1 .~\g: .~ft~c!( Y: t A~J. iJ: C'.~ll1I)~:litiOll ~,illt ht.xahydr~cark)linu~; t ) Rh ( a 1 P;2; and (Ll t B3. C: ( ' e m ~ t i ~ m with phc'nothktzine~: l b ) 22:1 = ! ("3: t {.;!) ('5; f l l (%: am~it • t ('7

, |ll~>lltp,.:lol¢ {II the ;tbkC~C~: o{ i~|!oli~activa~;;~hlc fig;rod ibl~ill~ cc~htrtttisL al!er [thttl ~};d ¢tfi.~.7 t,,ilh ibm: t~.~peftiw;" iig~tfld l[tt;llehCdk ;il;tt aftt'l 9ht.~fRslz~b¢tli~t ill tltc' t~fcsctl¢~ ol ~ ~l M ~l}il;~'fi)ll~2 lcl'os~;h~id~edk The rc~uti~ a ~ ~k~- m~an v~t!ut-s lff 1h177 imk-Nnd~ml exr>efime=lla t a c k peTti~rmed ie triplicate.

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m~xlttrc o{ h i t c h - a f f i n i t y D . a n l a e o n i s t s stlch a s { 4. )-buta~dg~m-o] ( e } a n d ~,ftipc~(mc ( a } ;~o-lcc~cJ a). ~cce?~.c-;'~ {ram'l ~¢~ ~{~*Ld~ ! D a , >-hdc l h c imtc~i~.e c n a n t i o m e ~ ~ -}-bmacLam~_d { } a n d ffic sdk-'c1:}v,c D~ ~'v'c,c{~¢~* awA,~,eo~'A04 S ( ' } l Z:33911 { z. } h~:d ~¢** ~'a~m:c1~'.c clk'c*

is irreversible but the c o m p e t i n g t g a n d interacts reversibly with the receptor. A s a furlher test of the specificity of the photorcaclion, each of the pho¢oacl~r a t a b l e iigands was prcphotoiysed (according to our standard protocol, see Materials and methods seclion} prior to incubation with m e m b r a n e s and reportm~ ligand. This treatment did not affec~ the potency of competitively inhibiting the binding of tile reporting ligand suggesting that the conditiotas of pholoactivation only affected the azido function but no{ those parts of the molecules invotvcd m reversible binding to d o p a m i n e receptors, Fig. 4 is a pharmacologic;:d!y rhOrC detailed rvq~reo scnlation of the specificity of photoa[fil~ity :[ah.c|Iilg o~ D., rcceptors l'~y com|~)tt/lds B3, C6 aml C7. |tcac the protection from labelling of D , sties was tested tic .a set of d o p a m i n e r g i c ligands of d:isd~,ct pi~armacoloD', Only De-specific ligands but not D,-speci~ic ,or~es. nor inactive i ~ m e r s , were c a p a b l e of protcction~ s:uggcs~i~lg that the photoreaction was specific,

radioactive k ~ b c i t ~ g w i t h higl~, - ~ - ' ~ - ...... ~'~. . . . . . . . . . mJucd h e d e t e r m i n a t i o n offi~,c re~,Aona~ !ocaiizau, o~n i~'i .1he '~crtebrate ce~traI nervous syslem of ti.~esc re.ccp,'