CA2189634A1 - Combinatorial dihydrobenzopyran library - Google Patents

Abstract

Combinatorial libraries are disclosed which are represented by the Formula (I): (T'-L)q- S -C(O)-L'-II' wherein S is a solid support; T'-L- is an identifier residue; and -L'-II' is a ligand/linker residue. These libraries contain dihydrobenzopyrans of formula (II) which interact (i.e., as agonists or antagonists) with .alpha. adrenergic receptors, dopamine receptor, .sigma.-opiate receptors, and K+ channels and are inhibitors of carbonic anhydrase isozymes. They are useful in the treatment of ocular diseases such as glaucoma.

Description

~ WO 95/30642 r~ 2 ~ 8 9 6 3 4 P~ C~
TITLE OF T~3E INVF~ITIQN
COMBINATORIAL DIHYDROBENZOPYRAN LIBRARY
BACKGROUNDOFTEJF IN\~FI~TION
There is interest i~, methods for the synthesis of large 5 numbers of diverse compounds which can be screened for various possible physiological or other activities. Techniques have been developed in which one adds i~1dividual units sequentially as part of the chemical synthesis to produce all or a ~ number of the possible compounds which can result frc~n all the different choices possible at 10 each s~qllPnti~l stage of the syn~hesis. For these techniques to be s-l~c~ccfi-l, it is necessary for the compounds to be amenable to methods by which one can li~otPmfin~ the structure of the compounds so made.
Brenner and Lerner (PNAS USA 81: 5381-83 (1992)) and WO
93/20242, for example, describe a synthesis wherein oligonucleotides 15 are produced in parallel with an~, are chemically linked as genetic tags to oligopeptides as the compoun~is of interest. WO 93/06121 teaches methods for particle-based syntllesis of random oligomers wherein n~ifi~tion tags on the particl~s are used to facilitate i~Pn~ifi~ri~n of the oligomer sequence synthesized. A detachable tagging system is 20 described in Ohlmeyer et al., Proc. Natl. Ac~d. Sci. USA, 9Q~ 10922-10926, Dec. 1993.
SUMl~ARY OF THE INV~TIQN
The present invention relates to ~lllbilldl.JIidl chemical libraries of compounds encoded with tags and to the use of these 25 libraries in assays to discover biologically active compounds. The present invention also relates to libraries containing dih~dlube~ )yrans and using these libraries to identify biologically active members by screening for inhibition of carbonic anhydrase isozymes. The present invention also relates to members of the library 30 which interact (i.e., as agonists or antagonists) with c~ adrenergic receptors, dopamine receptors, ~J-opiate receptors, and K+ channels. In particular, the present invention also relates to members of the library

2 ~ 5i, t ~ 2 1 8 ~ 6 3 4 r.~ v37~o which are inhibitors of carbonic anhydrase. The invention also relates to methods for their preparation, int~rm(~ t~, and to methods and ph~ l formulations for using these dih~d~h~llzo~yl~lls in the treatment of m~mm~l~, especially humans.
Because of their activity as inhibitors of carbonic .ydl~se isozymes, compounds of Ihe present invention are useful in the treatment of such diseases as glaucoma.
DETAILED DESCRIPTION OF THE INVENTION
The COl~ la~olidl libraries of the present invention are I~ sc.lltd by Formula 1:
(T'-L)q- (~) -C(O~-L'-II' wherein:
(~) is a solid support;
T'-L- is an identifier residue;
-L'-II' is a ligandllinker residue; and q is 3-30.
Preferred compounds of Formula I are those wherein:
T'-L- is of the Formula:
~ O--(cH2)n =C~l \0 111 wherein n = 3-12 when Ar is pentachlorophenyl and n = 3-6 when Ar is 2,4,6-trichlorophenyl;
q is 4-12; and 2 1 89 63~
95130642 '-`' ' ~

-L'- is ~O or ~NoB2~
~a) (b) wherein the left-hand bond as shown is the point of to the solid support and the right hand bond is the point of ~ hm~n~ to the ligand, and B is O or N(CH2)1 6R17, with the proviso that in (b) when B is N(~H2)1 6R17, the ligand is attached to B through a carbonyl group.
Othe} preferred ~ oullds of Formula I are those of For,nulae Ia, Ib, or Ic wherein -C(O)-L'-II' is:

O~ ~Rs Ia ~CH2)1 Rl? ~Rs Ib or (CH2) l 6RI7 ~Z\~\O--~Rs lC

WO 95130642 ' 2 i 8 q 6 3 4 ~ c~
Depending on the ch~ice of L' (see Table 1), the ligands of Formula 11 may be detached by photolytic, oxidative, or other cleavage techniques. For example, when -L'- is (b) and B is O (or N(CH2)1 6R17), photolytic detachment may be IG~lGS~ Gd by:
I (T l-L)q~3C(O)-LII + II'OH (B = O) or + II'NH(CH2), 6RI7 (B = N(CH2)~ 6RI7) wherein L" is the residue from ;~' and II'OH (or Il'NH(CH2)1-6Rl7)is II.
Therefore, compouL(ls of the pres~nt invention are also GSelllGd by Formula II

Rl--~'~ 3Rs R2~ o~4 wherein:
Rl is OH, O(CH2)1 2OH, OCH2CO2H, CO2H, o-Z-C(o)NH(cH2)l-6Rl7~ or OCH2-~Phe-C(O)N~J(CH2)1 6R17;
1~ R2 is H or lower alkyl;
R3 is H, alkyl, aryl, or arylalkyl;
R4 and RS is each independently ~1, lower alkyl, or substituted lower alkyl where the s~lhstit~lpntc are 1-3 alkoxy, aryl, s--hstit--t~d aryl, carboalkoxy, carboxamido, or diloweralkylamido; or R4 and Rs taken together are -(CH2)n-, -(CH2)2-O-(CH2)2-, -cH2-o-(cH2) -(CH2)2 NR8-(CH2)2-, -CH2 NR8-(CH2)m~, -(CH2)2CH(NHR8)(CH2)2-, -(CH2)2-S(O)0-2-(cH2)~-, or -CH~CH(I~-loweralkyl)(CH2h~HCHr;

WO 95/30642 ~ ~ " ~ 2 1 8 9 6 3 4 ~ .c lc one of R6 and R7 is H and the other is H, OH, or N(CH2) 1 6R14R15; or R6 and R7 taken together are 8 s s o s o, o S\ / ,or S\ /S ' R8is H, COOR9, CONHR10, CSNHRIl, COR12, So2R13, S loweralkyl, aryl loweralkyl, h~e.uLlyl, or heteroaryl lower alkyl, wherein aryl is optionally s--hstitlltl~d with 1-3 Sll~ llf ~ i selected from lower alkyl, lower alkoxy, halo, CN, NH2, COOH, CONH2, carboalkoxy, and mono- or di-lower alkylamino and wherein heteroaryl is a mono- or bicyclic h~ ualu-lldlic ring system of S to 10 members including 1 to 3 hel. lu~Llullls selected from O, N, and S and 0-3 substituents selected from halo, amino, cyano, lower alkyl, carboalkoxy, CONH2, and S-lower alkyl;
R9 is lower alkyl, aryl, aryl lower alkyl, heteroaryl, aryl s~hstit-~t~d by 1-3 substituents selected from alkyl, alkenyl, alkoxy, methylene dioxy, and halo, or a 5 to 6-membered heterocyclic ring wherein the hetero atom is O or N, wherein heteroaryl is a hetelualul.latic ring of 5 to 6 members including 1 to 2 heteroatoms selected from O, N, and S and 0-2 s~hstituPnt~ selected from lower alkyl, dialkylamino, lower alkoxy, and halo;
R10 and Rl I is each independently lower alkyl, aryl, aryl lower alkyl, or aryl sub~ u~L-d by 1-3 substituents selected from lower alkyl, halo, alkoxy, and haloalkyl;
25 R12 is lower alkyl, aryl, heteroaryl, aryl lower alkyl, hf t~Ludlyl lower alkyl, a 5- or 6-membered heterocyclic ring c~nt~ining 1-2 hL~Iud~ùllls selected from O, S, and N, a S- or 6-membered heterocyclic ring containing 1-2 ll~t~lu~ullls selected from O, S, and N lower alkyl, or aryl sl~h~titlltPd with 1-3 sllhstit~lfnt~ selected from lower , . ... .. . .. . ... . . .... ... ... . .. . .. .. ....... .... ....

WO95/30642 ~ ,n ~ ~ 2 1 89634 PCT/US95/05940 ~

alkyl, alkoxy, halo, sulfamoyl, loweralkyl sulfamoyl, cyano, and phenyl;
R13 is lower alkyl, aryl, or aryl ~ rd with 1-3 sl-hstit~lf ntc selected from lower alkyl, alkoxy, halo, CN, and haloalkyl;
5 R14 is H; alkyl ~u~ilL~,d by 1-3 alkoxy, S-loweralkyl, sulfamoyl, halo, alkylsulphonamido, or arylsulphonamido;
alkenyl; alkynyl; aryi; substituted aryl; heteroaryl;
~uL,~ilul~d heteroaryl- heterocycloalkyl;
~H2NR16C(O)R16; -C(O)NR16R16; -CH20C(O)R16; or ~H2SC(O)R16;
R15 is H, alkyl, -C(O)X, -C(S)X, or-C(NCN)NR3R3;
R16 is lower alkyl, sl-hctifllttod lower alkyl, aryl, or substituted aryl;
R17is H; alkyl ~ rd by 1-3 alkoxy, S-loweralkyl, sulfamoyl, halo, alkylsulphonamido, or aryl~"ll,h. "A",i~
alkenyl; alkynyl; aryl; sllhstitlltfd aryl; heteroaTyl;
il".~ d heteroaryl, heterocycloalkyl;
-CH2NR16C(o)Rl6; -C(O)NR16R16; -CH20C~O)R16; or CH2SC(O)R16;
X is alkyl, aryl, arylalkyl, O-loweralkyl, or NR3R3 Z is -(CH2)1 6-, optionally s--hctih-tf d with 1-3 lower alkyl;
CHR2; Phe-cH2-~ where Phe is optionally mono-~",1.~,~ illllf d with halogen, lower alkyl, or alkoxy; or heteroarylene-(CH2)-;
mis 20r3;
n is 4-9;
or a phar n~e~-ti~lly acceptable salt thereof.
Preferred compounds of Formula 11 are those wherein R12 is sulfamoylphenyl, most preferably p-sulfamoylphenyl.
A preferred embodiment of the invention is a compound of Forinula 1I wherein-Rl is OH, oCH~C(o)NH(CH2)1 6R14, orOCH2-~Phe-C(O)NH(CH2)1 6R14;
R2 is H or lower alkyl;

WO 95/30642 ~j ~ r, ~ ~ ~. 2 1 8 9 6 3 4 F~~ 710 R4 and R5 is each lower alkyl; or R4 and R5 taken together are -(CH2)s-, -(CH2)2-O-(CH2)2-, -(CH2)2 NR8-(CH2)2-, -(CH2)2-CH(NHR8)(CH2)2-, -(CH2)2-S-(CH2)2-, or -CH2CH(~NCH3)(CH2)2Cl HCH2-;
S R6/R7 are H/OH, =O, or -S(CH2)2S-;
R8 is H, COOR9, CONHR10, CSNHRIl, COR12, So2Rl3~ .
Iower alkyl, aryl lower alkyl, heteroaryl wherein the ring members include I to 3 N atoms and the substituents are halo or amino, h~t~,lu~lyl lower alkyl wherein heteroaryl is 6-membered and the h~,t~luatollls are N, or aryl lower alkyl sllh~titl~t~d with 1 ~ f ,l selected from lower alkyl, alkoxy, and halo;
R9 is lower alkyl, aryl lowe} alkyl, aryl, tetrahyd}ofuranyl, tetrahyd}opyranyl, o} aryl substituted by 1 to 2 ~ selected from lowe} alkyl, alkenyl, alko~y, methylene dioxy, and halo;10 and Rl l is each in~rren i~ntly aryl, aryl lower alkyl, or aryl substituted by 1 ~UIJ~ U~llL selected from lowe} alkyl, halo, alkoxy, trifluoromethyl, and pentafluoroethyl;0 R12 is lower alkyl, aryl, aryl lower alkyl, heteroaryl lower alkyl wherein the heteroatoms are N, a 5- or 6-membered heterocyclic ring cont~inin~ 1-2 heteroatoms selected from S and N lower alkyl, o} aryl ~ul~Liluled witn I ~ l ;l "~ ., selected from lower alkyl, alkoxy, halo, sulfamoyl, cyanû~ o} phenyl;
R13 is lowe} alkyl, aryl, or aryl ~--hstit -t~d with I sub~liLu~
selected from lowe} alkyl, alkoxy, and halo;
or a pharm~e--tic~lly acceptable salt thereof.
~lost preferred compounds of the invention are IG~ llt~d by the formula:

^i:
WO95/306 2 ': .' ? ~ ' S 2 ~ ~ 9~34 r~ 5 ~ R6 R~
Rl--~, Rs R2 Ila wherein:
Rl is 6- or 7-OH;
R2 is H or lower alkyl;
5 R4 and R5 is each methyl; or R4 and RS taken together are -(CH2)5-, -(CH2)2 0-(CH2)2-, -(CH2)2 NR8-(CH2)2-, -CH2-NR8-(CH2)3-, -CH2 NR8-(CH2)2-, or -(CH2)2-CH(NHR8)(CH2)2-;
one of R6 and R7 is H and the other is OH or R6 and R7 taken together are =O or -S(CH2)2S-;
R8is H, COOR9, CONHR10, CSNHRIl, COR12, So~R13, benzyl, -CH2-Ph4-F, -CH2-Ph-4-OCH3, -cH2-4 n-butyl, -CH2-c-propyl, ~ ~ ~N ~ ~ =S , or ~ ~ cCI;
R9 is i-propyl, phenyl, phenethyl, t-butyl, ~ ~ > ~0 R10 is phenyl, p-chlorophenyl, or p-trifluoromethylphenyl;
Rll is phenyl, benzyl, or l-naphthyl;

WO95/306~2 ~ C 21 89634 r~"~ 4^
R12 iS
~CN 0~5NyO ~;NH -CH2~3 H H
~3~ ~S02NH2; and R13 is 1- or 2-naphthyl, phenyl, ~chlorophenyl, ~
methylphenyl, ~t-butylphenyl, n-butyl, or i-propyl;
S or a plld~ lly acceptable salt thereof.
Most preferred compounds of the invention are also represented by the formula:

Rl--~ R5 lla wherein:
Rl is 6- or 7-OH when R2 is H
Rl is 7-OH when R2 is CH3 R4 and R5 is each methyl; or R4 and RS taken together are -(CH2)s-, -(CH2)2 0-(CH2)2-, -(cH2)2-NR8-(cH2)2-, ,-CH2-NR8-(CH2)3-, -cH2-NR8-(cH2)2-~ or-(cH2)2 CH(NHR8)(CH2)2-;
one of R6 and R7 is H and the other is OH or R6 and R7 taken together are =O or -S(CH2)2S-; and R8 is WO95/306~2 ~ t ~ r~l,. '^- ^

~ o~ ~--H J~' [~
~Y~ H2N02S~
~o~o~ o H ~S~
~~ o~c~'L' Cl{~So~
--~ ~ [~ ~ Me~O~
~ ~S S~ ~2~

WO 951306~2 ~ ., t ~ 2 1 8 9 6 3 4 PCTIUS95/05940 ~N--8 N~--C~ H3C~ 2 5 Cl--aH--8~ ~f 8 H C>--2 F3C~ ~, ~ t-Bu~
~N 1-13CO~/ H2N~
N Cl ~-- N~ N~--Cl~ ,~,, F~
D--~ t- BU~oJi~S, or H.

Most preferred compounds of the invention are ~ s~llt~d by the formula:

Rl--~Rs R2 lIa wherein:
S Rl is 6- or 7-ocH2c(o)NH(cH2)l-6Rl7~ or 6- or 7-OCH2-4-Phe-C(O)NH(CH2) 1 6R17 when R2 is H;
Rl is 7-OCH2C(O)NH(CH2)1 6R~7, or 7-OCH2-4-Phe-C(O)NH(CH2) 1 6R17 when R2 is CH3;
R4 and RS is each methyl; or R4 and R5 taken together are -(CH2)s-, -(CH2)2-0-(CH2)2-, -(CH2)2 NR8-(CH2)2-~
-(CH2)2-CH(NHR8)(CH2)2-, -(CH2)2-S-(CH2)2-, or -CH2CH(jNCH3)(CH2~2CI HCHr;
or R4 is methyl and RS is C~20CH3 or -(CH2)3N(Et)2;
one of R6 and R7 is H and the other is OH~ or R6 and R7 taken together lS are =O or -S(CH2)2S-; or one of R6 and R7 is H and the other is NAB, where A is methyl, 2-methoxyethyl, 2-phenylethyl, 4-methoxybenzyl, 2-tetrahydro-furanylmethyl, 2(3,4-dimethoxyphenyl)ethyl, or 2,2-diphenylethyl and B is H, -SO2C~3 . ~OJ~ ' ~
~, (CH2)4NH2 ~ (CH2)sNJ~ NH2 C~ t ~ 2 1 ~ 9 WO~S/31)642 634 r~-~u~

NCN O O
N(n-propyl)2 ~ ~ NHCH3 ~ ~ NHPh S S
~ NHCH3 or ~ NHPh R8 is H, CONHCH3, S02Ph, (CH2)3CH3, CO(CH2)2CH3, benzyl, -C(0)-(4-Phe)-S02NH2, or N=\
N~ -~13 is O O
H, -S02CH3, ~ oJ~ ' ~
(CH2~4NH2 ~(CH2)5NJ~ NH2 NCN O o ~N(n-propyl)2 ~ ~NHCH3 , ~NHPh S S
~ NHCH3 or ~ NHPh ~
25 (CH2) 1 -6R14 is methyl, n-butyl, 3-methoxy-n-propyl, CH2-c-propyl, or -(CH2)1 3-phenyl; and (CH2)1 6R17 is methyl, 2-methoxyethyl, 2-phenylethyl, 4-methoxybenzyl, methyl-2-tetrally-llu~u.anyl~
2(3,~dimetho~yphenyl)ethyl, or 2,2-diphenylethyl;
30 or a pharmaceutically acceptable salt thereof.

~`` ? ~ 21 8 9 6 3 4 WO 95/30642 P~ ,.3.'C'. I^
-1~
Especially preferred. as inhibitors of carbonic al~tly~llas~.
are compounds of formu~ae ~b, ~c, ari~ ~d.

Rl--~N-R8 ~ ~N R8 llb lIc Rl ~LN~R8 lId wherein:
Rl is 6- or 7-OH, 6- or 7-oCH2C(o)NH(cH2)l-6Rl7, or 6- or 7-OCH2~Phe-C(O)NH(CH2)1 6R17;
R2 is H or CH3;
R8 is -CO-Ph-p-SO2NH2; and R6 and R7 together are =O or -SCH2CH2S-.
Most preferred of these are the following compounds:
Formula Formula lIc Formula lld llb R1 7-OH 6-OH ~OH 6-OH 7-OH

R6/R7 o -SCH2CH2S- -O- -SCH2CH2S- -SCH2CH2S-R14 and R17 may each be any pharmq~olo~irqlly relevent orgaic radical, such as those de~ived by removal of H2NCH2- from the following compounds:

~ WO 95/30642 ~ C ~ ~ 8 9 6 3 4 r~l~u~ ~c ~

CH
H2N-CH2--CH~ ~C CH3 H2N-CH2-cH2~", H

H2N-CI~12 ' CH \H2 H2N-CH __~

O HN
H2N-CHz~CH2~CH2 N\ Cl (~ CH2 H2C~ NH2 2~ H2N -CH2 ~ CH~
H2N \CH3 CH--CHz--CHz--CHz--CH2--NHz / ~CH2--NH
H3C--O--C~ o ~ NH HzC--CHz ~,CH2 H2N-CH2-CH2--N/ \

WO g5r306~2 2 ~ ~ 9 ~ 3 4 PCTIIIS95105940 ~
HzC--CH2 2 CH2 CH2 CH2 N~
H2N CH2-CH2 NH~N ~ H2N CH
H2N--CH2 ~ , / C~
2C H2N-CH2-cH2~ H2N--CH2 ~CI
H N--CH ~> H2N-cH2~>
H ~C

H2N--CH2~ ~3~-C~2 [~

~ ~ Ir~ ~1 2 ~ 8 9 6 3 4 H2N-CHZ (~ ~ H3C~CH2~NH2 H N
0 z ~CH2 H2N CH2 ~ r F
H2N_CH2~CI H2N CH2 ~--Cl H C NHz

3 ~ H3C O~CHz-NH2 H3C--O~CH2-NHz 3 ~ \ ~CH2--NH2 H2N~ F\C/F
2~ F H2N-cH2--~ F

f~ ~S ~89634 WO95130612 r~ J.. ,3,v~j~0 ~-~'~) /cH - c~ ;

H3C--Cl--CH2-NH2 H3C--CH2 NH2 ,CH~
H3C~ ~CH3 2C o,CCH~o H2N-CH2-CH2--NH~>
,CH2~ ~CH2~

2~c~ ~ /C--NH_CH
HzN
H~C
,CH2 CH--NH_CH

2 ~ 8 9 6 3 W095~30642 -19- 4 r~
H3C~o H3C--NH--CH2-CH2--NH2 ~\
H3C--o~cH2-cH2-NH2 H3C-CH2-CH2--NH--CH2-CH2-NH2 H2N--CH2--CH2 \~ / rBr 1~ H3C--O--CH2-CH2-NH2 H2N-CH2-CH2 ((~ F
20 HZN-cH2-cH2~ H2N--CH2--CH2 rC
H2N--CH2-cH2~
2~; H3C--O~CH2-CH2-NH2 NH
30 H3c~o CH~CH~ ~CH2-CHz-NHz 2 1 8 9 6 3 ~
W0 95/306~2 F ~ ~ t ' ~c H2N--CH2--C=CH H3C CH2-CH

H2N--CH2 CH=CH2 H3C--CH2--0--CH2-CH2--CH2-NH2 CH
1~ H3C--C--CH2--CH2--NH2 H2N-CH2-CH2-cH2 ~ H2N-CH2-CH2-cH2-cH2 25 ~H--CH ~ H3C CH2 CH2 CH2 CH2 NH2 H3C--CH2 CH2 NH2 H3c-cH2-cH2-cH2-cH2-cH2-NH2 t ~ r ~, ~ 2 1 8 9 6 3 4 ~ wo 95/30642 -21- r ~
H3C-CHz~CH2~CHz~CH2~cH2~cH2-NH2 H2N-CH
F
H2N-CH2-cH2~ ~ =~F
IC . H31c ~15 H2N_CH2-CH2-CH2 N~ H2N-CH2 ~ ~ C--F

20 ,CH2~/ ~F <~C--F
2~ H2N-CH2~ \--F ' Cll2`C~
o CH,~ 'F H3C-CH2-cH2-cH2-NH2 ~' ~ S 2 1 ~ 9 6 3 4 WO95/30612 r.~ Oi~0 HzC--C.~ H N_CH2 2--~C--'~-H2 CH2 ,~
10 CH CH \ ~,CH2 CH2CH2NH2 r\\'~\1 F ~ ~ ~, CH2-CHz_NH

a~ ~Br H2N-CH2--< '~ ' ~H3 CH~ ,o' ~

` ;.'~ `. ~189634 wo 9s/30642 r~ J., .'C~q 1^

CHz`~"~

CHz~ CH/
HZh Z CH ~\ ~ o/C~c 1~ Z ~ H3C,CHZ NHz H2N - CH2 ~Br NHz H2N--CH2 ~ F--C--F
z-_ N~/ ¦

3~ 89634 WO 9S/30642 -24 r~
H2N-CH2-cH2~ HZN-CH2 _H2-CH2 Cl CH2-CH2 ; ~C~
\ O N CH2 2 CH2 --\\ H3C CH
0 2~NH2 15 H3C--CH2--S--CH2--CH2--NH2 H2N-cH2-cH2-cH2 N~ CH
Cl H2N--CH2--C CH ~ CH -NH
Ol F F F
O/c~ H2N-CH2--C--C--C--F
H3C cH2~NH2 F F F

H2N-CHz-CH2-CHz--Br O B
CH2-oH2 \(~ , W0 95/306~2 ~ t ~' ' 2 ~ 8 9 6 3 4 r~
-25- .
H2N CH2--CH2--S~ ~ H~C-CH2-CH2-CH2-CH2-CH2--O--CH2-CH2-CH2-NH2 H2C~
H3C--S--CH2-CH2-NH2 H2c-o~cH2-NH2 ~o 15 H3C--CH2--S--CH2-CH2-NH2 H2N-CH2-CH2-CH2-cH2 C\ \H2 //o H2N-CH2--C~
20 H3C--CH2-CH2--CH2--S--CH2--CH2--NH2 ~ / ~--Br H3C--O--CH2-CH2--O--CHz-CHz-CHz-NHz /~\ ' - HzN-CHz~

~ ~ ,(" ~ 2 ~ 89634 WO 9~/30642 i ' ~

H3C~CH2_NH2 ,CH~
10~'~ CH2-NH2 H3C--CH2--O--CH2--CH2 NH2 15H2N-CH --~-- CH3 CHZ-CH CH
C\H3 C\H3 O Cl CH3 20,CH2 ~ ,CH~
H3C 1l `CH,~NH2 ;~N-~/ 2~NH H C--CH -CH2--CH2--O--CH2-CH2--CH2 N 2 30 ~ , 2 ~ , 2 ~ 89634 WO 95/30642 ~ `. r~ .'c~, l^

~\ CH2 CH2_CH

~3Co H \H~H2 H3C~ ~H2 ~; H,C--o~ N~ N.

H2C--C ,~

_CH2-CH2 CHz A
25 H2N ~ H2N-CHz~
N~

WO 95/30612 ~ F~ 10 ~0 0~
C ~C CH
,~ 2 CH2 CH2--CH2--N 1 2 0 o CHz 5 ~ l\ H2~NH2 H3C~ ~_CH -CH2-NH2 o H2N
l l C~H2 ,c~ 'IH2 , CH CH2 H3C_N~
H3C \__N
I ~ CH2 ~/ CH2-CH C ~ --CH--C\H
~/CH2-CH ~N' ~wo 95r30642 ~ 2 1 8 q ~ 3 4 . ~ s ,^

~CH, ~ CH~ ~ -- Z CHz_NH
H2N CH2 CH2~N H2c-cHz-cH2-cH2-cH2-cH2-cH2-cH2-NH2 H C-CH2-CHz-CH--0~ CH ~ ~>

H3C-CHZ-CH2-CHZ-CH2-CH2-NH2 H~-CH -CH ~--O--CH2_~
H C CH CHz 2 CH2_CH2_CH CH3 H N C _~
Cl r~ ~C 2189634 wo gs/30642 ~ r~

N ~ NH \ CHI ~C--H H_CH_C/
- CH~
g~ oN~o~
O N~

\C/ CH2`NH ~

~ ~ 1l 20 ~~ '' 'H2 ~ NH2 a~, O
H,C--CH2~ r / . a~2--Q~-~H2-CH2--NH2 ' CH2 CHz~ H \\o ~ C

0~ ~O~ ~,C~

2 t 8 9 6 3 4 WO 95/30642 T.,IIL

~ H3C-CH2-CH2--O--CH2-CH2-NH2 s H2N-CH ~Br H3C-CH2-CH2-CH2--O--CH2-CH2-NH2 H2N-CH2 / ~Cl 2 CH2-CH ~Cl HzN-CH2~ O I H2N-CH2-CH2 `~ ~rI
~ 0 c_cR2-NH2 r~

~2 ~2~o H3C-CH2--O--CH2-CH2--D--CH2-CH2-CH2-NH2 \c~~~
. _ . . .

,~ 2t89634 WO 95/30642 ~ C -32- r~-~u~ C
\C--NH--CH2-cH2-cH2-cH2-cH2-cH2-NH2 CH2~ ~
H3C '~ 1~2 H2N NHz 2 ~"Cl CH~
CH2~,CH3 HZN-C

H C---~CH2-NH2 H2N-CH2--C--r--F

CH2 HzN-CH2--I--l C--C--~--~--C--F
H3C 2~NH2 2 ~ 8 9 6 3 4 ~ c~ ~c H2N-CH2 = Z
0 HZN-cH2 ~ F H2N-cH2-cH
/ CH2--CH HzN-CH2-CHz--S--CHz~
H3C/ 2 cl CIH3 Cl H3C ~CH2,NHZ HzN-CHz-CHz--S--G~z~
25, ~_H3 H2N--CH2----NH
CH~ NH2 CH/2 2 H3C` ~,CH2' H C-- C ro CH, WO95/30642 ~ ' ' F~,1/L .

~--NH.~ C CH2_CH

C--NH~ CHz~cH
0H2N 2~NH2 15; H2N CH2~0~ /
20 ~ 0~a~-~ H2N-C~2-CH2 H3C C~C~
CH3 ---N~//O NHz H3C 0~CH2-CH2-NH2 ~N-C1 12 C~

Br \~\ ~ NH2 H ~ 2 CH2 \\
H3C--CH2 NH2 H N-cH2-cH2-cH2 1 O 2 HC~N
CH3 H2\ \
1~ H~C--I--S--CH2-CH2-NH2 / CH2_NH
C~ \Cl H2N--CH2--CH=CH2 ~ _o~
H2N--CH2-CH2--~cl H~ CH-CH. ~-a~
Hf--O~O--C~1z-CH2 ~H~ H2N C~2 1 I F

W0 95130642 . 2 1 8 9 6 3 4 iF~~

N lH2 H3C~o I ,CH3 H2N`-CH2-CH2 0~
0 2 ~ CH2`C~ Cl 3C~o~CH2 H2N_CH2~ ~o cCI 3 H N--CH2~Cl H2N-CH2--C_C~
cH3C
H2N--CHz~CI C~ ~ C\,CH2NH2 Cl CH3 ~ Cl ~C~

t.
wo ss/30642 2 ~ 8 9 6 3 4 r~ J

H2N 2 ~/ ~0 ~_CHZ-cH2~
0 N~(~__ NH2 \\C
HzN-CH2-CHz-CHz-CH--NH--~CI C--NH--CH ~ ¦
~ NH~ ,~/ / \0~

~ z-~z-a~z-c~z-c~z~-~--s~ 3c~o~ CH2 ,NH2 H2N-CHz-CHz-CHz-~Hz--NH--~ H3C CH~ H2 .

W0 95130642 ~ ; r~

b H2C--CHz H,C--0~CHz~CH2~NH2 H2N--CH2--CHz--c ~ CH2 o H2C--CH2 3 \ ~0/ CH2 H2N--CH2-CH2-CH2--CH \CH2 , ~ CHz~ HzC--CHz H2N_CH HZC CH
Z CH2_NH // H2N--CH2--CH2 CH~

~\ ~ `CH--CH3 20 CH-~HZ- z \o ~c~ CH2 C~ 2-cH2 ~ H C-CH2-CH2 ~H2 ,C~ CH
\\

r ~ t ~
~W095/306~2 2189~34 P~

HzN-CHz-CH2~ C~'2_c ~

H2N--CH2--CH2~CI H3r-~ ,CH2 H3C-CHz--O~CH2-CH,-NH, H2N-CH2-CH2~Br ~~ r ~ C~2 ~ 3 F NH
'~

HZ~I--CH2~0--C--F CH2~F
F

WO95/30642 ~ ` 2 1 89634 P ~

HzN-CH2-CHz~CI H2N--CH2--CH2--<0 H2N--CH2 CH2~ CH _NH~
Cl H3C--C/~H H3C ,~CH2-cH2-NH
CH--CHZ-CHz-CHz-CHz--NHZ O l \\
,CH~ H3C--CH2-cH2--NH2 CHz CH
H3C 2~
~C,cH3 H2N~ F
H3C CH2~,~/F
HzN
CHi H2N--CH2 ~ C--F

1~ t ~ 2 ~ 8 9 6 3 4 WO95130642 ~ F~IIIJ '~' I^
~1-H2N~ " ,F
H2C H~ C;C--cH N CH _CH2_NH \\ \~ Cl 5 CH2--CH ~ H C ~ ~-->
20 H2N--CH2 CH2 )~ ~ ~ CH _NH--~/o 25 ~CH,-O--\~ CH2-'H2-NH2 , _~

~c~ H3c-cH2-NH2 r~ r ~ ~n ~ ~ 21 89634 H2N~ ~C C C~ --C\

C~Z`CH~ ~ CH2~c 20~,-c~-c~ C7~2 c~ c~ CH _NH--[\ CH
25H~N-CHz-CH2-CHz-CH2-CH2~ ~c~ , ~O\H3 NH~ H2N-CHz~

WO 95/30642 ~ ; 2 1 8 9 6 3 4 P~
-43- . -H
3C H ;~
H3C-CH2--o~CH2 CH2-NH2 \\f--S_C~Z~ Cl ~C~-CHz-c~ H3C--11--CH2--CHZ--CH2--NH2 H3C--CH2~CH2-CH2-NH2 ~~

2~ Hjc ~c, ~-~z-c~-c~z~ ~\Cl CH/
~ ~0 F
H2N-CHz-CHz~O~ H2N-cH2-cH2-cH2--S~

i '~' J 1' ~ 2~ ~963~
WO 95/306.12 r~ 9 o H2N--CH2 ~ CH2 ~ j CH
0 ~D--C--F H3C--O--CH2-cH2-cH2-cH2-NH2 H2C--CH2 H2N--CH2 C 2 \ ~c12 H3~C
Cl'/ 2----CH ~) - ~J' I CHz-CHz-NHz [~
25H~N-CHz-CH~--CH~-CH2-CH~--CH~--CH~--o~) CHz--f~
C, ~ o ~
H N-CH2--CH2-CH2--S C\ ~ ,_ CHz \

2 ~ 8 9 6 3 4 WO 95/30642 - P~ '.'C', IC

C~, ~;~C~2-CH~ 2 2 CHZ
5c~
0 ~5~ \~ Cl~O--CH2~(~
5 5 ~ CH2 HzN CHz CH3 \\ ~C--~
20 CH _S-- NH_CH/ CH2 H3C--CH2--CH2--O--CH2-CH2--CH2 NH2 2N 112C~ ,CH, CH ~ `CH \i~, ~ CH2~CH
zN H,C--~ Cl 2~NH2 ~H
-CH ~ C~2~CH2~

~r ,` -` f ~t1~; 21 89634 r~l,.J. )~~~ l~

H3C~o \~C--NH--CH,-CH2-CH2-NH2 H2N--CH2-CH2--0~ ~ ~0--CH~
S
H3C H2N- 2 ,~
H3C_o @ ~ H2'NHZ H2N 2 Xo N
H2N-cH2-cH2 S~ I H2N--CH2--CH2--~
Cl~--~H2~ F ~C--F
CL~z H2N-CH2-cH2 // --CH2 o ~O
~ H NH--CH2--cH2--NH2 H3C--O~cH2--cH2-NH2 Cl CH
Cl H3C

t ` ~ ~ 8 ~ 6 3 4 W095130642 ~ r.

~-CH2-ch,~N~ Cl 2 , ~ ~1 H2N` ~
Z\ ~ H2N-CH2-CH2 ~ : ~ ~F

~ ~H2` ~`~9 H3C CH2 ~ 01 HN R,C N~
25~ H2~ ~2-~ ~) H2N--cH2-cH2~ r ~ 2 CH2~

WO95/30612 ~ t'~ 2189634 r ~u~

CH3~sCH3 H OY
H3C-(CH2)7CHCH(CH2)~NH2 H3C-(CH2)9-NH2 H3C-(CH2)12-NH2 ' ,0 <C~
H3C-(CH2)1~-NH2 c~
(Cf~2,~
(CH2~4NH2 2~; H3C-(CH2)1~NH2 o~Nl~f (cH2)4NH2 0 H3C ~H~o,O(CH ~ N~

n tS 2~8963~

H N-CHZ-CH2~ 2--CH=C/

~N H2N CH2 CH2 Cl -- \CH--CH2 Cl l2 H C--o--l~o CH ~CH2~
CH3 ,c~7. ~f H3C Cl CH3 C~3 J~

H2N ~ F~ --F
~Ch _NHZ \\C--NH--~

S ~ 1 89634 W0 951306 12 . ~ I / 1),. ~ '~ I ^
-~0-c' F
H3C~ ~CH CHz H2N_CH ~B~
' H3C~ cHz_l~H~
15 ~; ~\`F CHz 20 ~ C~ H3C--CH2-cH2-cH2-NH2 25 H N-CH ~) F H2N-c~y~Cl ~ ~Br ,CH2 ~2.~H H2N

WO9!i/30642 u ;~ i,'C 2 ~ 8~634 p~ c NH2 @~
o o o ~ \\ cH~CHz \CH2 O--NH O
15H N_CH2--~ ~0 CH-CH2-CH2 2 2~

CHz H,N~-0~2~--11l!~ H c C /

~o ~ H_~ O-t2 2 ~, ~-CH2-a~ff~,-~H,-C~U--11~ CH _CH2 \Ci' c 2l 8963~
W095/30642 ~ 52- r~ o~ Ic H2N--CH ~ ~ ~ CH ~~
~ f~ H2N--CHz--CH2-CH2--cec~
~
~J NH~ C ~ C~

Cl ~ H3C--O~CH2-CH2-NHz ~ I ~ CH2 ~
~/ CH2`CH

`. S,~ ~! Q ~'C`.
WO95/30642 2 ~ 89~34 r~-~u~ 4~ -~C~-~ --C\

Cll ~ B CH2~CH
0 h3C 1~ N 2 5 ~--~H~, ,r~2 I~iH~
H~'l H3C_r~ ~ f' H2N--CH2-C ~`--F
~ ~~f H2 ~ ~ i CH2 ~ ~\r~~ F
HzN-CH2 ( ~ ~ ~Br b~

2 t 89634 WO 95/306~2 ~ r.

NH
H3C-(CH2)l1-NH2 c=o H3C O~
H3C C~12 5 ~ ~N
(CH3)3NH2 .

WO 9S/30642 r~~ ,S't One embodiment of the invention is the use of the combinatorial library of Formula I in assays to discover biologically - active compounds (ligands) of Formula 11. Thus, an aspect of the invention is a method of identifying a compound having a desired - 5 characteristic which comprises 5yntht~ci7.ing, a combinatorial library of Formula I and testing the compounds of Formula I and the ligands of Formula II, either attached to the solid support or detached therefrom, in an assay which identifies compounds having the desired characteristic. A further embodiment of the invention is determining the structure of any compound so identified.
Another embodiment of the invention is a process for preparing a compound of the formula:

3 ~~ ~OLH
where R2 is H or lower alkyl;
which comprises a) reacting allyl or methyl ~(hydroxymethyl)-3-ni~lùb~ ual~ with a compound of the formula:
o HO--~ Me in the presence of triphenylphosphine, toluene, and DEAD and stirring the mixture at room tr~ atul~ to produce R2C`~O ~ Me where R is allyl or methyl and b) when R is allyl reacting said compound with methylene chloride, tetrakistriphenylphosphine p~ lm(0), and pyrrolidine and stirring , . . . . .. , . . .. . .. . . . .. . . . . _ _ _ i~ 21 89634 WO 95B06~t P~~

the mixture at 0 C, or when R is methyl reacting said compound with dilute NaOH and THF and stirring the mixture at 0 C.
Forthis reaction, R=allyl is preferable to the t-butyl or methyl esters since the milder con-litionc would not induce aldol type S c-~n-i~nC~ n of the a~t;lupllc. olle portion of the molecule.
Another embodiment of the invention is a method for identifying ~u~ oullds that are inhibitors of carbonic anhydrase which comprises preparing a mixture of 20-300 pmol test compound and aqueous solutions (total volume: 25-100, preferably about 50, IlL) of 10 0.03-0.12, preferably about 0.06, ,uM carbonic anhydrase and 0.04 0.16, preferably about 0.08, IlM d~ ylalllide7 exposing said mixture to U.V. (preferably 274 nm) light, and d~ g the amount of emitted U.V. ~preferably 454 nm) light.
Another embodiment of the invention is a method for lS identifying compounds that are enzyme inhibitors which is a lawn assay which co.u~.is~s cont~ctin~ a colloidal matrix c ~"~i.i";"~ enzyme, which matrix has ~mherirll~d therein a mono-layer of solid supports with attached ligands, with a layer of fluL~Ivgt;llic substrate-co-ltdillillg material, eluting said ligands by exposure to U.V. Iight, and detecting 20 zones of inhibition in the colloidal matrix produced thereby. A
preferred such lawn assay comprises cont:~fin~ an agarose matrix cr~nt:~inin~ bovine carbonic al~l~ydl~ with a llu~ ceill diacetate-cont~inin~ layer of agarose.
Another embodiment of the invention is a compound of the 25 formula:
Br o N2 )l /~R
(~N)~ ~ `(CH2)l~RI7 wherein:

WO 95/30642 2 r~l",~

~. .
Is a solld support;
R is H or alkyl;
- R16 is lower alkyl, sllh~itllted lower alkyl, aryl, or substituted aryl;
R17 is H; alkyl substitute~ by 1-3 alkoxy, S-loweralkyl, sulfamoyl, halo, a~kylsllirhon:~mi(ln, or arylsulphonamido;
alkenyl; alkynyl; .~ryl; substituted aryl; heteroaryl;
~llhstitll~ed heteroarYI; heterocycloalkyl;
-CH2NR16C(O)R16; -C(OiNRl6Rl6; -CH2OC(O)R16; or CH2SC(O~R16; an~
Y is aryl or heteroaryl.
Compounds of forrn.nla 14 are useful as i~ l-.cli~tcs in the construction of combinatorial libraries and are especially useful in automated or batch mode syntheses thereof.
5 Definitio~s The following abbreYiations have the indicated meaning:
Boc = t-butyloxycarbonyl c- = cyclo DEAD = diethylazodicarboxylate DBU = 1,8-diazabicyclo[5,4,0]undec-7-ene DCM = dichlolomethane = methylene chloride DIC = diisopiopylcarbodiimide DMAP = 4-N,N dimethylaminopyridine DMF = N,N-dimethylformamide DMSO = dimethyl sulfoxide DVB = 1 ,~divinylbenzene EDT = 1,2-eth~nP~iithi equiv. = equivalent Et = ethyl FACS = fluorescence activated cell sorting Fmoc = 9-fluorenylmethoxycarbonyl GC = gas chromatography HOBt = N-hydroAyb~--,vlLid~ule hr = hour, hours ~ ~ ., 111 t ~ 2 1 ~ 9 6 3 WO 95/30642 ' r~ a.

im = imidazole in = indole m- = meta Me = methyl 5 Mtr = 4-methoxy-2,3,6-trimethylhen7~n~s--1fonyl n- = norrnal Naph = naphthyl p- = para PEG = polyethyleneglycol Ph = phenyl Phe = ph~llylc;ll~
Pmc = 2,2,5,7,8-pentamethylchroman-6-sulfamoyl py = pyridyl r.t. = room temperature sat'd = saturated s- = secondary t- = tertiary t-Boc = t-butyloxycarbonyl TFA = trif~uoroacetic acid THF = tetrallydluruldll "Alkyl" is intended to include linear, branched, o} cyclic structures and combinations thereof of from 1 to 20 carbon atoms.
"Lower alkyl" includes alkyl groups of from 1 to 8 carbon atoms.
Examples of lower alkyl groups include methyl, ethyl, p}opyl, isopropyl, butyl, s- and t-butyl, pentyl, hexyl, octyl, c-propyl, c-butyl and the like. "Lower cycloalkyl" includes cycloalkyl groups of from 3 to 8 carbon atoms. Examples of lower cycloalkyl groups include c-propyl, c-Dutyl, c-pentyl, 2-methylcyclopropyl, cyclopropylmethyl, and the like.
"Alkenyl" is C2-C6 alkenyl of a linear, branched, or cyclic (Cs-C6) configuration and combinations thereof. Examples of alkenyl groups include allyl, is~lu~ lyl, pentenyl, hexenyl, c-hexenyl, 1-propenyl, 2-butenyl, 2-methyl-2-butenyl, and the like.
"Alkynyl" is C~-C6 alkynyl of a linear, branched, or cyclic (Cs-C6) cnnfiEI~inn and combinations thereof. Examples of alkynyl groups include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl W095/30642 ~ 2 ~ ~d q~34 r~",J. ~c l^

isopropynyl, pentynyl, hexynyl, c-hexynyl, I-propynyl, 2-butynyl, 2-methyl-2-butynyl, and the like.
"Alkoxy" means alkoxy groups of from I to 6 carbon atoms of a straight, branched, or cyclic configuration. Examples of 5 alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy, and the like.
"Substituted loweralkyl" means lowe~ alkyl ~ub~ilul~d with 1-3 alkoxy, carboalkoxy, call,u~a-llido, di-loweralkylamino, aryl, substituted aryl, or heteroaryl.
"Aryl" means phenyl or naphthyl.
"S--hstit~t~ d aryl" means aryl ~ul~liluled with 1-3 halo, loweralkyl, alkoxy, aryl, S-loweralkyl, alkylsulphonamido, arylsulphonamido, or sulfamoyl.
"Heteroaryl" means a 5 or 6 membered aromatic ring 15 containing 1-3 hetero atoms selected from O, N, and S.
"S-~hstitl-t~d heteroaryl" means heteroaryl s~bstitllt~d with 1-3 halo, loweralkyl, alkoxy, aryl, S-loweralkyl, alkylsulphonamido, arylsulphonamido, or sulfamoyl.
"Heterocycloalky" means lower cycloalkyl containing 1-3 20 hetero atoms selected from O, N, and S.
Halogen includes F, Cl, Br, and 1.
L and L' are depicted in Table I, which also shows cleavage reagents. In designing a synthetic scheme, L and L' are chosen such that they are orthogonally reactive; i.e., they must allow for 25 removal of either T or Il (where T = T'-OH) without removal of the other since ~".".ll~,-rous cleavage of both T and 11 from the solid support is &sadv~nt~o~c. In the structu~es as shown, the left-hand bond is the point of ~tt~t~hm~nt to the solid support and the right-hand bond is the point of ~tt/~hm~nt to either T or 11.

WO 95130642 ~ t~ 2 1 8 9 6 3 4 r~ c TABLE I
LINK-~R GROUPS
Linker Group Cleavage Reagent NO2 hv 1............ ~CH2B- or CH2O~LB-~\ hv 2. 02N~ CH20-,=<OR Ce(NH4)2(NO3)6 3. ~0-~\ Ce(NH4)2(N03)6

4. RO~o s. --Cl:l=CH(CH2)2- 03, 0sO4/104-. or KMnO4 6 --CH=CHCH2- 03, 0sO~/104-, or KMnO4 7 --CH2CH=CH- 03, OsO4/104-, or KMnO4 2G 8. ~;~ 2) H30+
9 . --CH=CHCH20- (Ph3P)3RhCl(H) Br Li, Mg, or BuLi 10. ~0-11- --S-CH2-O Hg+2 Zn or Mg 12. I CH2-O- ~
OH Oxidation, e.g., Pb(OAc)4 13. I CH2-O- or H5106 30 R = H or lower alkyl X = electron withdrawing group such as Br, Cl, and 1.

p ~ ~ ~ 2 t WO95130642 - ~9634 ~ c~ ,^

The tags of this invention, T, are chemical entities which possess several properties: they must be detachable from the solid supports, preferably by photolysis or oxidation; they must be individually dir~ lidble, and preferably separdble; they must be

5 stdble urlder the synthetic con~itinn~; they must be capable of being detected at very low con~nt~ti~ n~ e.g., 10-18 to 10-9 mole; they should be identifiable with readily-available equipment which does not require sophisticated technical capabilities to operdte; and they should be relatively economical. The tags may be structurally related or 10 unrelated, e.g., a homologous series, repetitive functional groups, related members of the Periodic Chart, different isotopes, co,--l,i,-alions thereof, and the like At the end of the ~ol,l~ dluridl synthesis, to each solid support, there will usually be attached at least 0.01 femtomol, usually 0.001-50 pmol, of each tag. The tags may be aliphatic, 15 alic~clic, aromatic, heterocyclic, or combinations thereof.
D;~ ";~ features may be the number of repetitive units, such as methylene groups irl an alkyl moiety; alkyleneoxy groups in a polyalkyleneoxy moiety; halo groups in a polyhalo compound; -and/or ~ s..l)~ d ethylene groups where the substituents may be 20 alkyl groups, oxy, carboxy, amino, halo, or the like; isotopes; etc.
The materials upon which the c~,---bil-dL~rial syntheses of the invention are performed are referred to as solid supports, beads, and resins. These terms are intended to include:
a) beads, pellets, disks, fibers, gels, or particles such as 2~ cellulose beads, controlled pore-glass beads, silica gels, polystyrene beads optionally cross-linked with divinylbenzene and optionally grafted with polyethylene glycol and optionally f ln( tir)n~li7l-d with amino, hydroxy, carboxy, or halo groups, grafted co-poly beads, poly-acrylamide beads, latex beads, dimethylacrylamide beads optionally 30 cross-linked with N,N'-bis-acryloyl ethylene diamine, glass particles coated with hydrophobic polymer, etc., i.e., material having a rigid or semi-rigid surface; and b) soluble supports such as low molecular weight non-cross-linked polystyrene.

.~ 2 1 89634

6.12 ~ /i5~ 10 ~

lt is intended that the definitions of any ~ ;l",-."l or symbol (e.g., R3) in a particular molecule be independent of its definitions elsewhere in the molecule. Thus, NR3R3 ~ ~ NH2, NHCH3, N(CH3)2, etc.
5 Optical Isomers - Di~ctf reomers - Geometric Isomers - Tautomers Some of the compourlds described herein contain one or more aùy~ lic centers and may thus give rise to el~sn~ir rnPrc~
liaù~-c;olll~,lù, and other stereoisomeric forms which may be defined in terms of absolute stereochemistry as (R) or (S). The present invention 10 is meant to colll~ ld all such possible dia~ olll~l~ as well as their racemic and optically pure forms and mixtures thereof. Optically active (R) and (S) forms may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic double bonds or other 15 centers of geometric asymmetry, and unless specified otherwise, it is intended to include both E and Z geometric isomers. Likewise, all ~dU~OIl.C~ic forms are intended to be included.
Salts The ph~rm~e~l~ic~l compositions of the present invention 20 comprise a compound of Formula 11 as an active ingredient or a pl.,.l ,~ lliC~lly acceptable salt thereof, and may also contain a p~ ic~lly acceptable carrier and, optionally, other ~ ld~uiic ingredients. The term "ph:lrrs~ell~ic~llly acceptable salts" refers to salts prepared from ph~ c llic:~lly acceptab~e non-toxic acids or bases 25 including organic and inorganic acids or bases.
When a compound of the present invention is acidic, salts may be prepared from pl-~""~r~ llic~lly acceptable non-toxic bases.
Salts derived from all stable forms of inorganic bases include ~ minllm, ~mmf~ni~lm calcium, copper, iron, lithium, m~nt-cillm, 30 m~ng:~nl~Se, potassium, sodium, zinc, etc. Particularly preferred are the ~...",(~,~i",.., calcium"""~ "~ 1-, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, sl~hstitl~ted amines including naturally occurring ùlll~illllPd amines, cyclic amines 35 and basic ion-exchange resins such as arginine, betaine, caffeine, ~ WO95130642 ' ~ t 8 9~34 r~ .a~ 5 IA

choline, N,N'-dibenzylethyli ..r l; ,~-;"P, diethylamine, 2-diethylaminoethanol, 2-dimethyl~min-ethsnol, ethanolamine, ethyl, nPrii~minP N-ethylmorpholine, N-ethylpiperidine, g~ minP
~ co-,llll;ll~ histidine, isopropylamine, Iysine, methylglucosamine, 5 morpholine, pi~ dLil~C, piperidine, polyamine resins, proeaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, etc.
When a eompound of the present invention is basic, salts may be prepared from pharm:~re~-tic~lly acceptable non-toxie acids.
Such aeids inelude aeetic, t~ lllfonic, benzoic, r~mrhorclllfonic, 10 citrie, eth~nPc~lfonie, fumaric, gluconic, glutamic, hydrobromie, hydroehlorie, isethionic, lactic, maleic, mandelic, mP.thqn.oclllfonic, mueie, nitrie, pamoie, pantothenie, phosphoric, suceinic, sulfuric, tartaric, p-toluenesulfonie, etc. Partieularly preferred are eitrie, llydlul)luLIlie~ maleic, phosphoric, sulfuric, and tartaric acids.
In the ~lic( llcQion of methods of treatment herein, reference to the compounds of Formula II is meant to also include the ph~. "~ c~lly :~cept~hlp salts thereof.
Utilities The ability of the c~ lpuun~ of Formula Il to interact with a adrenergic receptors indicates that the compounds are useful to treat, prevent, or amelioate lly~ t~,~ion and benign prostate hypertrophy in m~mm~lc, especially in humans.
The ability of the compounds of Formula 11 to interaet with dopamine reeeptors indieates that the eompounds are useful to treat, prevent, or :~mPli~ tP Alzheimer's disease and depression in humans.
The ability of the GGIllpoulld~ of Formula 11 to inteaet with ~J-opiate reeeptors indieates that the compounds are useful to treat, prevent, or :~m~linrltP sclli~u~,l-l- llld in m~mm:llc especially in humans.
The ability of the coLupoul~ds of Formula Il to interact with K+ channels indieates that the eompounds are useful to treat, prevent, or amelioate hypertension, astnma, and pulmonary insufficiency in mzlmm~lc, espeeially in humans.
The ability of eertain eompounds of Formula Il to inhibit earbonic alLllyd-dse isozymes makes them useful for p~ve~ g or reversing the symptoms induced by these enzymes in a mammal. This .. . .

WO 95/306~2 ~ ; 2 1 8 9 6 3 4 Pi~ 'J ~ ~ ~

enzyme inhibition indicates that the compounds are useful to treat, prevent, or ameliorate ocular diseases, particularly glaucoma in mz mm~lc especially in humans.
Dose Ran,aes The msi~nitlld!~ of the prophylactic or thP-, re-ltin dose of the co~ Jc ullds of Formula 11 will vary with the nature and severity of the condition to be treated and with the particular compound of Formula Il and its route of ~ , l inn In general, the daily dose range for anti-en~ymic use lies in the range of 20 to 0.001 mg/kg body weight of a mammal, preferably 10 to 0.01 mg/kg, and most preferably 1.0 to 0.1 mg/kg, in single or divided doses. In some cases, it may be necessary to use doses outside these rarlges.
When a comrncitinn for intravenous adlllil~ lion is employed, a suitable daily dosage range is from about 10 to 0.0005 mg (preferably 5 to 0.01 mg) compound of Formula Il per kg body weight When a composition for oral ~ (,,.lion is employed, a suitable daily dosage range is from about 20 to 0.001 mg (preferably 10 to 0.01 mg) compound of Formula 11 per kg body weight.
When a composition for ophthalmic adminis.~ration is employed, a suitable daily dosage range is from about 10-0 01%
(preferably 5 0-0.5% compound of Formula II, typically prepared as a 2.0-0 1% by weight solution or ~ ioll of a compound of Formula Il in an acceptable ophthalmic formulation The compounds of Formula 11 may also be used in combination with other rhz rm~ellticz lly active ingredients. For example, a typical ocular r~" "",l~li"" may comprise the compound alone or in combination with a ~-adrenergic blocking agent such as timolol maleate or a paras~",~ ,..",i"~r~ic agent such as pilocarpine.
When used in combination, the two active ingredients are present in 30 approximately equal parts.
Phzirm~celltin~l Compositions Any suitable route of administration may be employed for providing a mammal, especially a human, with an effective dosage of a compound of Formula 11. For example, oral, rectal, topical, parenteral~
35 ocular, pulmonary, nasal, etc. routes may be employed. Dosage forms ~ WO 95130642 ~I f' ~ f~ -r~ 2 1 ~ q ~i 3 ~ r~

include tablets, troches, ~i~y~ iulls, sllcrenci~nQ solutions, capsules, creams, ointments, aerosols, and the like.
The yll~l,llacrulical compositions of the present invention comprise a compound of Formula 11, or a plld~ r~ rAlly acceptable 5 salt thereof, as an active ingredient, and may also contain a ` pharm~rellti~llly acceptable carrier and, optionally, other ,~ Ally active ingredients.
The compositions include compositions suitable for oral, rectal, topical (including ~AI~ ",~1 devices, aerosols, creams, 10 ointments, lotions, and dusting powders), parenteral (including ~ub_u~ncous, ;..I.,..."~ 7r, and ill~ldv~nuus)~ ocular (ophthalmic), pulmonary (nasal or buccal inh~l~ti~n), or nasal A,~ l;On;
although the most suitable route in any given case will depend largely on the nature and severity of the condition being treated and on the 15 nature of the active ingredient. They may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of phall--a.y.
A compound of Forrnula 1I may be combined as the active ingredient in intimate admixture with a phd~ r~ul;~Al carrier 20 acco-ding to conventional ph~ r~ l cu...~uu--di--g techniques. The carrier may take a wide variety of forms ~IPpPn~lin~ on the nature of the ylt;~f l~ desired for ~ minictr~ion i.e., oral, parenterdl, etc. In preparing oral dosage forms, any of the usual ~ . Il.A~r--l ;~1 media may be used, such as water, glycols, oils, alcohols, flavoring agents, 25 pl~,se.~ali~i" coloring agents, and the like in the case of oral liquid prçr~ mc (e.g., suspensions, elixirs, and solutions); or carriers such as starches, sugars, microcrystalline cellulose, diluents, ~rAnlll~ing agents, lubricants, binders, flicintPgrAtin~ agents, etc. in the case of oral solid prep~rAti~lnC such as powders, capsules, and tablets. Solid oral 30 pl~pdlaliO~5 are preferred ove} liquid oral prepardtions. Because of their ease of ~,I---; -i~l~AI;~-n, tablets and capsules are the preferred oral dosage unit form. If desired, capsules may be coated by standard aqueous or non-aqueous ~Pf;hniq~lPc - In addition to the dosage forms described above, the 35 compounds of Formula 1I may be ~f1minictf~red by controlled release means and devices such as those described in U.S.P. Nos. 3,536,809;
.. . .. ... ....

WO 95/306~2 ~ 8 P~ll.J~,." I ~

3,598,123; 3,630,200; 3,845,770; 3,916,899; and 4,008,719, which are incorporated herein by reference.
Pharm~ellti~l compositions of the present invention suitable for oral a~ on may be prepared as discrete units such 5 as capsules, cachets, or tablets each containing a preAPt~rrninf~d amount of the active i..~ in powder or granular form or as a solution or sl~crencion in an aqueous or nonaqueous liquid or in an oil-in-water or water-in-oil emulsion. Such compositions may be prepared by any of the methods known in the art of pharmacy. In general, the compositions 10 are prepared by uniformly and intimately admixing the active lC;diCII~ with liquid carriers, finely divided solid carriers, or both and then, if necessary, shaping the product into the desired form. For example, a tablet may be prepared by coll.l,.c~;on or molding, optionally with one or more accessory ingredients. Cul~ ~cd tablets 15 may be prepared by CUIllL~ illg in a suitable machine the active ingredient in a free-flowing form such as powder or granule optionally mixed with a binder, lubricant, inert diluent, or surface active or Aicr~ rsin~ agent. Molded tablets may be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert 20 liquid diluent. Ophthalmic inserts are made from culllpl~s~ion molded films which are prepared on a Carver Press by subjecting the powdered mixture of active ingredient and HPC to a compression force of 12,000 Ib. ~gauge) at 149-C for 1-4 min. The film is cooled under pressure by having cold water circulate in the platen. The inserts are then 25 individually cut from the film with a rod-shaped punch. Each insert is placed in a vial, which is then placed in a humidity cabinet (88%
relative humidity at 30 C) for 2-4 days. After removal from the cabinet, the vials are capped and then autoclaved at 121-C for 0.5 hr.
The following are IC~ IlLltiv~ pharm~lrelltic~l dosage 30 forms of the compounds of Formula 11:

~ WO 95/30642 .';, s Ç '~ ~ ~ 2 1 ~ 9 6 3 4 r~.,~J,,,~.t~ l^

I.M. ~nj~rt~hle Suspension mglmL
Compound of Formula 11 10 Methylcellulose 5 - Tween 80 o.5 S Benzylalcohol 9 B~n7~1konillm chloride Water for injection to a total /o~ume of 1 mL
Taklet m~ltablet Compound of Formula Il 25 MicrQcrystalline cellulose 415 Povidone 14 Pl~gc~ rd starch 43.5 M~nPcillm stearate 2.5 Capsule m~/r~pc.. l,~
Compound of Formula 11 25 Lactose powder 573.5 M~n!~cillm stearate 1.5 Aerosol Per canister .
Compound of Formula 11 24 mg Lecithin, NF liquid concentrate 1.2 mg Trichlorofluoromethane, NF 4.025 gm DichlorQdifluoromethane, NF 12.15 gm Ophth~lmi~ Solution ~BL
Compound of Formula 11 Monobasic sodium rhocrh:~t~-2H2O 9.38 Dibasic sodium phosphate l2H2o 28.48 BPm:~lkoni~-m chloride Water for injection to a total volume of I mL
Ophthalmic Sus~enSiQn Compound of Formula 11 Petrolatum liquid to a total weight of 1 g WO 95/30642 ~ 2 1 8 9 6 3 4 Qphthalmic Insert m~/insert Compound of Formula II
Hydroxypropylcellulose 12 These compounds of Formulae I and I~ may also be used as 5 libraries for di~7COV~ g new lead structures by evaluation across an array of biological assays, including the discovery of selective inhibition patterns across isozymes. These libraries are thus tools for drug discovery; i.e., as a means to discover novel lead compounds by screening the libraries against a variety of biological targeLc and to 10 develop structure-activity relationships in large families of related ~o~ oullds. The libraries may be tested with the ligands attached to the solid supports as depicted in Formula I or the individual compounds II
may be detached prior to evaluation. With the compounds of Formula I, screening assays such as FACS sorting, bead lawn assays, and cell lawn 15 assays may be used. When a compound is detached prior to evaluation, its IGldliUll.711i~ to its solid support is ,,,~;,,li.i,,r~l for example, by location within the grid of a standard 96-well plate or by location of activity on a lawn of cells. The solid support ~cco~ t~d with bioactivity or the solid support related to the detached ligand may then be decoded 20 to reveal the structural or synthetic history of the active compound (Ohlmeyer et al., Proc. Natl. Acad. Sci. USA, 90, 10922-10926, Dec.
1993).
Assays for Det~ ninin~ Biolo~ical Activity The compounds of the present invention may be tested by 25 assays well known in the art for interaction with a adrenergic receptors, interaction with dopamine receptors, int~.~rtion with a-Qpiate receptors, interaction with K+ channels, and carbonic anhydrase inhibition. For example, I~ ~-c..e~ liv~ }eferences teaching carbonic alll.ydl~se inhibition assays are:
Carbonic Arlhydrase Inhibition - Maren and Couto, The Nature of Anion Inhibition of l{uman Red Cell Carbonic Anhydrases", Archiv. of Biochem. a~.~d Biophy., 196, No. 2, Sept., 501-510 (1979).

S 2 1 8~634 W0 95130642 ' P

Carbonic AnhYdrase lnhihi~isn - Ponticello et ~., "Thienothiopyran-2-sulfonamides: A Novel Class of Water-Soluble Carbonic Anhydrase Inhibitors", J. Med. Chem., Q, 591-597 (1987).
Carbonic Anhy lr;~P Inhibition - It has now been found that 5 the use of very low initial c-)nrPntr~ti-~ns (0.04-1.6, preferably about 0.6, IlM) of dansylamide and (0.03-1.2, preferably about 0.3, ~IM) of carbonic anhydrase to assay test compounds for carbonic anhydrase inhibition not only allows the use of very small total volumes (approx.
25-100, preferably about, 50 IlL) per assay but also allows one to 10 distinguish high-affinity from low-affinity compounds without either re-elution or re-synthesis of the test compound. By i.~ asing the c,~ rl,~ rinn of ~la~ ldlllide from ~0.1 ~IM to -200 ,uM directly in the assay sample, relatively weak inhibitors can be distinguished from relatively strong inhibitors on the same aliquot of test compound. The 15 small total volume advantageously permits high throughput assaying of small quantities of test COlllpuull~i~, for example, in 96-well plates, and the reduced c~n~ n of dd-l~ylal--ide advantageously permits the detection of test compounds that have a wide range (<500 nM) of .lr,~ lir~coci~ >n constants. The following materials are used:
100. mM sodium phosphate buffer, pH 7.4 0.6 ~M dansylamide (Sigma D-3882) 0.3 ,uM bovine carbonic anhydrase (Sigma C-3934) inhibitor Reactions are carried out in 50 ,uL total Yolume in 96-well plates, preferably, Dynatech MicroFluor plates, white with 'U' bottom, c~nt~inin~ the test compounds. The assay mix is prepared immP~ tPIy before use, and 50 ,uL of the assay solution is pipetted into each well of 30 plates in which the test compounds are previously dried. The plates are spun briefly in a tabletop c."li~iruge before reading fluorescence.
Fluorescence is read in a Perkin-Elmer LS 50B spectrofl~ rimPter fitted with a Well Plate Reader Accessory using an excitation wavelength of 274 nm (2.5 nm slit) and an emission wavelength of 454 35 nm (20 nm slit), with a 390 nm cutoff filter in place. ~luorescence Illen~u,t:lllc~ are averaged over 1 sec for each well. To identify wells in which inhibitors are present, first a plate with no exogenous .3i~,C 2189634 WO 9~/30642 ~ J,,,S.'C

inhibitors is read, which typically gives a fluorescence reading of 2.
3.1 (typical standard deviation i 0.06) for a given assay solution. In plates CUll~dillillg inhibitor ~ntli~ Pc, active inhibitors cause a decrease in the lluulci~c~,.lce signal of greater than S times the standard deviation.
S To di~ uish high-affinity from low affnity c~n~ t~c, 5 ,uL of a 2 mM stock of dansylamide in DMSO is added to the above test solution, and the assay repeated as above. Typical readings are 7.5 to 8.5iO.4 (standard deviation) among previously identified inhibitors.
High-affnity compounds lower the signal by greater than 3 standard deviations). Thus, the increased conr~n~r~tic n of dansylamide is sufficient to displace relatively weak inhibitors (e.g., chlorothiazide, Ki - 75 nM) without displacing relatively strong inhibitors (e.g., dcGld~ula,~,ide, Ki ~ 7.5 nM).
Bead Lawn Assay (General ~ethod). An enzyme of interest is illcolluùlaLed into a gellable gum such as silica gel, agar, agarose, pectin, pOlya~ ylalllide, gelatin, starch, and gellan gum, preferably a low melting-L~ agarose gel (0.5-2.0%, wt./vol.), which is layered on top of a lawn, no greater than one bead in thickness, of solid supports with attached ligands. The detection of an active combinatorial library member is accomplished by photoeluting the ligands from the beads in slru by exposure to U.V. Iight. To minimize ~ la~ult;
photoelution, the beads are preferably protected from ambient light sources prior to U.V. exposure. The beads are evaluated by placing a second layer, preferably low-melt agarose gel, G.~ E a substrate on top of the one co,.li~.. ~ the enzyme and the photoreleased library members, and allowing enzymic CUllvtil:~iull of substrate into product by diffusion of the substrate into the enzyme-con~inin~ gel. The substrate is preferably one that produces a photometric change upon conversion into product; e.g., the generation of a colored product, a fluorescent 30 product, or a chpmilllmin~scpnr reaction (where one of the products is a photon). The second layer may comprise a gellable gum such as silica gel, agar, agarose, pectin, polyacrylamide, gelatin, starch, and gellan gum, or a solid material such as a matrix c..~-ti.i..;l,~ an array of fluorogenic-pellets. Inhibition of the enzyme by a library member 35 results in a difference in d,u~ lallcc in the vicinity of the attached bead and allows for selection of the bead and the identifiers which encode for 2~ 89634 WO 95/30642 ~ PCT/US95/05940 the inhibitor. This technique may be used with a variety of enzymes, for example:
Acid Phosphatase Furin Activated Protein C y-Glutamyltranspeptidase 5 Alkaline Pl,n~l,h,.l~c~ G~dl~yllles A & B
All~ plidases B & M HIV Protease Amyloid A4-G~ dli--g Enzyme IL-lB Convertase An~io~ sindse Kallikrein Aryl Sulfatase Lysozyme ~-G~ tnc~ c~ Mast Cell Protease 3-Glll~nc~ e Peroxidase 3-Glucuronidase Plasmin Calpains I & 11 Prohormone Convertase C~h~p~in~ B, C, D, & G rANP Precursor Processing Enzyme Cholin~st~r~ce Renin Chymotrypsin Spleen Fibrinolytic Plul~i-la~c Collagenase Staphylocoagulase Dipeptidyl Peptidases 1- IV Thrombin Elastase Tissue Pl~ li.log~l~ Activator Endothelin Converting Enzyme Trypsin Factor Xa Tryptase Factor Xla Urokinase Factor Xlla Df-Protease A bead lawn assay for testing carbonic anhydrase inhibition 25 preferably ~Ulllplis~s agarose for both layers, bovine carbonic anhydrase, and fluorescein diacetate.
Bead Lawn Assay (Carbnni~ Anhydrase). Beads to be tested are arrayed in a minimal amount of methanol in a 60 mm polystyrene tissue culture dish and then all the methanol allowed to 30 evaporate. A 2.5% (wt.lvol.) mixture of agarose (SeaPlaque, FMC
BioProducts, Rockland, ME) in 20 mM sodium phosphate buffer (pH

7.4) is heated on a hot plate until the agarose dissolves and then is equilibrated to 37C in a water bath. A separate stock of the same buffer is also equilibrated to 37C. The enzyme layer is prepared as 35 follows: 100 IIL of a bovine carbonic anhydrase stock (0.5 mg/mL or 53 ~M based on absorbance at 280 nm, Sigma #C-3934) is added to 2.15 mL of buffer, and 1.25 mL agarose solution is added to the mixture. The agaroselenzyme solution is poured onto the dish WO 951306S2 2 1 8 9 6 3 4 ~ C~

Gn"li~i";.,P: the beads and the agarose is allowed to solidify at r.t. for 3-5 min. To identify zones of inhibition, the compounds, which are optionally photoeluted by exposure to 4.7-6 mW/cm2 365 nm UV light for 5 sec. to I hr., are overlayed with fluorescein diacetate (FLDA, 5 Molecular Probes, Eugene, OR), which is prepared as follows: to 2.25 mL pllo~h~t~ buffer is added 10 ,uL FLDA stock (10 mM in DMF at -20C) and 1.25 mL agarose (final FLDA c~ "~.,l""ion 30 ~M). The solution is mixed thoroughly then poured over the enzyme layer in the dish. Zones of inhibition appear after 1-2 min. and intensify over 30-10 45 min. They are dark against a yellow-green background when illllmin~ d by short-wave UV light (~ ax = 254 rlm).
Bead Lawn Assay (Inositol Mo~ n~l)l."l,.~e). The assay is similar to that for carbonic anhydrase, with the following ~ub~liluLioll~. The buffer used is 20 mM Tris, I mM EGTA, pH 7.8.
15 The enzyme layer contains I mg/mL It;CVIIIbill~ human inositol monophncrh~ ce (purified from E coli) and 10 mM MgC12. Three alternative substrates are used: m~tllylulllbelliferyl phosphate (Sigma, M-8883), a nuOI~g~l~iC substrate, detected using filters around 7~ex =
388 nm and ~em = 420 nm; or CSPD or CDP-Star (rh~milllminPccPn~
20 sllhs~c for alkaline phosphatase, Tropix, Bedford MA), detected directly without requiring filters. The preferred substrate is CSPD.

2t 8~63~
~ WO95/30642 ~ ' r~ .o~ 1^

Methods of Synthesis The compounds of the present invention can be prepared according to the following methods. At each step in the synthesis each solid support upon which a compound is being synthesi~ed is uniquely tagged to define the particular chemical event(s) occurring during that step. The tagging is accomplished using irl~ntifie.rc such as those of Formula rv, which record the sequential events to which the support is exposed during the synthesis, thus providing a reaction history for the compound produced on each support. The i~l.o.ntifierc are used in ~G~ uind~ioll with one another to form a binary or higher or~er encoding scheme permitting a relatively small number of identifiers to encode a relatively large number of reaction products. For example, when used in a binary code, N ille.ntifi~r.c can encode up to 2N different compounds and/or cnntlitinnc By associating each variable or combination of variables at each step of the synthesis with a combination of i-l~nfifierc which uniquely define the chosen variables such as reactant, reagent, reaction Gon~1ition.c, or combinations of these, one can use the identifiers to define the reaction history of each solid support.
In car~ying out the syntheses, one begins with at least 103, desirably at least 104, and generally not exceeding 1015 solid supports.
Depending on the pre-determined number of RI/R2 choices for the first step, one divides the supports a.,c~ ,ly into as many containers. The d~ t; reagents and reaction corl~litinnc are applied to each container and the c~ tillll of itlentifiP-c which encode for each Rl/R2 choice is added and attached. Depending on the ch~mictri~c involved, the tagging may be done prior to, co-~o--~ ly with, or aher the reactions which comprise each choice. As a control, sample supports may be picked at any stage and a portion of their tags detached and decoded to verify that the correct tags are bound to the sample supports. As needed, one may wash the beads free of any excess reagents or by-products before proceeding. At the end of each step, the supports are usually combined, mixed, and again divided, this time into as many containers as pre d~,t~ -cd for the number of choices for the second step in the synthesis. This procedure of dividing, reacting, c WO 95/306.12 2 1 8 9 6 3 4 F~.ll.J:,. '~, ~^

. tagging, and remixing is repeated until the ~o.llbi~.dL~ l synthesis is completed.
Scheme 1 Functionalized supports such as amino-fi~n~ tion~1i7Pd or 5 hydroxy-lc,.llilldLi-~g PEG grdfted polystyrene beads are divided into a pre~ r. ,..;..Pd number of reaction vessels and are reacted with a cleavable linkerlligand element_, which has been pre-formed, to generate _. Unique tagging of the supports in each reaction vessel is achieved with combinations of identifiers encoded in a binary scheme, lO e.g., as depicted in Table 1-1 for three choices of Rl and R2. The j(1Pntifiprs are attached by adding a solution of the itiPntifiPr~ (in a 1.5%
wt./wt. identifier:solid support rdtio) to a batch of supports sllcpPn-l~d in CH2C12 and shaking the mixture for 30 min. A dilute solution of rhodium Llillu(~l~Jdct~ dimer is added and the mixture is immP~ tPly 15 shaken 4 hr and washed in CH2C12. The procedure is repeated and the mixture shaken for 14 hr and then washed in DMFIDCI~JI.
Scheme 2 The compounds _ are pooled, mixed, and divided into a pre-determined number of reaction vessels, each of which is treated 20 with one reagent corresponding to ligand element =CR4R5, in the presence of pyrrolidine to produce 5a, 5b, and 5c. Unique tagging of the supports in each reaction vessel is achieved with comhin~innc of additional i~iPntifiPr~ encoded in a binary scheme, e.g., as depicted in Table 1-2 for seven choices of R4R5-2~ Scheme 3 The colllpoul-ds 5c, where R4/R5/x I~ IL~ the residue of pipe~idine, pyrrolidine, or aminocyclohexane, are pooled, mixed, and then divided into a pre-determined number of reaction vessels. The supports in each reaction vessel are uniquely tagged with combinations 30 of additional if lPntifi~Prs encoded in a binary scheme, e.g., as depicted in Table 1-3 for 30 choices of R8 and in Table 1-5 for six choices of Rl4 and four choices of heteroaryl groups. After removal of any N-C 2 ~ 8 963~
WO 95/30642 PCT/I~S9!i/059~0 protecting Boc group in R4R5, each reaction vessel is treated with one reagent corresponding to ligand element R8 in the presence of solvents such as CH2C12, DMF, or EtOH and, when required, bases such as - triethylamine or 2,6-lutidine to produce 6 having an R8 s~ titllPnt at 5 C-2 and a ketone at C-4, i.e., when R6R7 together are O. In Scheme 3 R14 is ber~7yl, -CH2-Ph-~F, -CH2-Ph~OCH3, -cH2-~py~ n-pentyl, or-CH2-c-propyl; and h~ u~l yl is N~ ' N~ ' ~, or ~N~CI
Scheme 4 A portion of the compounds 5a, ~, ~, and 6 may be poûled, mixed, and then divided into a pre-determined number of reaction vessels where they may be uniquely tagged with comhin~tinnc of ~-lfliti-)n~l identifiers encoded in a binary scheme, e.g., as in Table 1-4 for three choices of R6/~7. Each vessel is treated with sodium borohydride to yield 7 as an alcohol at C~ or is treated with 1,2-dithio~thS~nP and a Lewis acid such as BF3-Et20 to yield 8 as a dithiolane at C4, or is treated with an dp~lulJ~idt~ non-beta branched primary amine in the presence of NaCNBH3 in MeOH, optionally with acetic acid, to yield secondary amine 9, or is left untreated.
Compounds 5a, ~2, ~, ~, k, 7, 8 ,and 9 are then exposed to UV light (~360 nm~ in polar solvents such as DMSO, H20, or a Iower alkanol such as MeOH to cleave the compounds of Formula 11 from the support/linker complex.
~cheme 5 TentaGel resin may be modified with bis-Boc Lysine to increase the available reaction sites for ligand ~tt~ hmPnt Bis-Boc-lysine in DMF, HOBt, and DIC are shaken at r.t. and then dry TentaGel resin is added. The mixture is shaken at r.t. for 17 hr and then washed alternately with methanol and DCM and then with THF and dried under vacuum. To deprotect the resin, DCM is added, followed by a 30 % TFA solution in DCM (100 mL). The vessel is shaken at room l~lllp~ldlul~ for 15 min. before adding neat TFA.. The vessel is shaken

8 9 6 3 4 WO 95/30642 2 1 PCTNS95/059~0 at room t~ d~UI~ for 2.5 hr at which time the resin is washed with DCM, then treated with a solution of 10% triethylamine in DCM, then washed with DCM and DMF.
For purposes of simplicity. the schemes do not show the use of this bis modification.
Scheme 6 Functionalized supports such as amino-~-n~tion~li7~d or hydroxy-lellllillaLillg PEG grafted polystyrene beads are divided into a pre-determined number of reaction vessels and are reacted with a cleavable linker/ligand element 10, which has been pre-formed, to generdte I 1. Unique tdgging of the supports in each redction vessel is achieved with combinations of idf~ntifi~rc encoded in a binary scheme, e.g., as depicted in Table 2-1 for seven choices of -(CH2)l 6R17. The identifiers are attdched by adding a solution of the identifiers (;n a 7%
wt./wt. id~ntifier:solid support rdtio) to each batch of supports 5-lcren.l~d in EtOAc and shaking the mixture for I hr. A dilute solution of rhodium ~linuuluac~L~te dimer in DCM is added and the mixture is shaken 15 hr and washed with DCM (4X) and EtOAc (2X). The ~lucedulci is repeated for each identifier.
To deprotect the encoded resin, it is suspended in DCM and then agitated with a TFA solution in DCM. The resin is then washed with DCM followed by treatment with triethylamine in DCM and then washed with DCM.
Scheme 7 The compounds L are pooled, mixed, and divided into a pre-determined number of reaction vessels, each of which is treated with one acetophenone reagent corresponding to ligand element R2, in the presence of DIC, HOBt, and DMF to produce 4'. Unique tagging of the supports in each reaction vessel is achieved with combinations of additional identifiers encoded in a binary scheme analogous to that in Table 2-1.
Scheme 8 The compounds 4' are mixed, pooled, and divided into a ~"~lt;L~ ed number of reaction vessels, each of which is treated with ~ W0 9S/30642 1. ~ 2 ~ 8 9 6 3 4 r~ r 1^
and aldehyde or ketone element cor~esponding to R4/R5 in the presence of pyrrolidine in methanol at 75C to produce the compounds ~, ~, and 5c'. Unique tagging of the supports in each reaction vessel is achieved with combinations of z~f-~tlitinn~ ntifil~rs encoded in a binary 5 scheme analogous to that in Table 2-1.
Scheme 9 The compounds 5c', where R4/R~/X l~?lc~??~ the residue of t-Boc protected piperidine, t-Boc protected aminocyclohexane, or other amine fi~nrtinn~li7~d molecules are mixed, pooled, and divided 10 into a ~ d~l~.lllil.ed number of reaction vessels. The supports in each reaction vessel are uniquely tagged with combinations of ~-lAitinn~l identifiers encoded in a binary scheme analogous to that in Table 2-1.
After removal of any N-protecting group in R4/R5, each vessel is treated with one reagent such as a chloroformate, iso, yalla~, 15 thioisocy2nate, carboxylic acid, alkyl or aryl sulfonyl halide, aldehyde, or a haloh~t~ loal.,u-dlic compound col~ .?l~dillg to ligand element R8 in the presence of solvents such as CH2C12, DMF, EtOH, or methanol.
When required, bases such as tri?~Lllylalllillc, DBU, or 2,6-lutidine and/or other reagents or combinations of reagents such as DIC, 20 NaCN~,H3, HOBt, and acetic acid are added to produce ~ having an R8 ??,~ ,t at C-2 and a ketone at C-4, i.e. when R6R7 together are O.
Scheme 1(`
A portion of compounds ~a~, ~, ~, and 6' may be pooled, mixed, and then divided into a pre-~ t~rminPA number of 25 reaction vessels where they may be uniquely tagged with combinations of ~r.~ itir~nz?l i('r.ntifirr.~ encoded in a binary scheme analogous to that inTable 2-1. Each vessel is treated with 1) sodium borohydride in met} anol to yield 7' as an alcohol at C-4; 2) 1,2-dithioethane and a Lewis acid such as boron trifluoride etherate to yield 8' as the 30 dithiolane at C-4; 3) an unhindered primary amine along witr.
NaCNBH3 in acetic acid/methanol solvent at cc~. 75C to yield 9' as an amine at C-4; or 4)is left untreated.

WO95/306~2 "i``~ C 21~9634 r~l~u~

Scheme 1 1 The compounds 5 or ~ are divided into a~ predetermined number of reaction vessels. Each vessel is treated with one reagent such as a chlolorullll~ , isocyanate, ~hioisocyanate, carboxylic acid, alkyl or aryl sulfonyl halide, aldehyde, ~r a haloh~elvdlullldlic compound corresponding to ligand element R15 in the presence of solvents such as Cf~2C12, DMF, EtOH, or methanol. When required, bases such as triethylamine, DBU, or 2,~1u~ic.ine and/or other reagents or combinations of reagents such as DIC, NaCNBH3, HOBt, and acetic acid are added to produce the corre~onding compound 13 or 13'.
Schemç 12 ~
Functionalized sur?orts such as amino-filn~ ti~n~li7~ or hydroxy-f~ . ,.,;.,~l;,,~ PEG grafte~ polystyrene beads are placed into a reaction vessel and are reacted with a cleavable linker/ligand element 10, which has been pre-formed, ~o generate 11'. To deprotect the resin, it is suspended in DCM and therl agitated with a TFA solution in DCM.
The resin is then washed with DCM followed by treatment with triethylamine in DCM and then washed with DCM to yield 12'.
In an ~p~ JIia~ly ~ized synthesis vessel is placed HOBt (3 equiv.) and the carboxylic acid ~? (X = OH) (3 equiv.) in a solvent such as DMF. DIC (3 equiv.) is added and the vessel agitated for 15 min.
before adding the amino resin 12' (I equiv. of amino sites). The resin is agitated for 5 hrs., then washed with ~If~rn~fin~ DCM and MeOH (SX
each) and then with THF (2X) to yield 14.
In an appropriately si_ed synthesis vessel is placed the amino resin 12' (I equiv. of amino sites). A solvent such as DCM is added, followed by an organic base such as triethylamine, pyridine, Hunig's base (di-isopropylethylamine), or 2,6-lutidine (10 equiv.). The resin is agitated for 15 min. before adding the acid halide Q (X = Cl, Br) (5 equiv.) as a dilute solution in a solvent such as DCM. The resin is agitated for 4 hrs. and then washed with DCM and MeOH (SX each) to yield 14.

W~95131~6~2 ' ~ 2~8963~ vcrrusgs/0s940 LINKERIlSt LIGAND ELEMENT
t-BuO~C~ HO--~Me OH
NO2 DEAD, PPh3, toluene t-BUOzC~o ~Me 2 DCMITFA ~O~ Me Identiders IVX ~o ~LMe 4 NO~ R2 OH
x is ]-30, depending on the binary code for the selected solid support WO 951306 ~2 ADDITION OF R4/Rs Pynolidine IV, H~
Lower alkanol Sa (I) NO2 R2 o Rs NO R~R5 (X is =CH2, or =O) ~ ~ ~Rs~
~X is =N(t-Boc) or =CHNH~t-Boc)) t ~;;;. 2 ~ ~ 9634 WO95/30642 r~ u.. ,',Jr I

f ~
ClCo2R9 2,6 lutidine l(lf~ntifif~r~/ O lo 50%TFA/DCM CNR
SCNRII
. / HO2RI2 d (n ~ ~ DlC, DMF
ClSo3RI3 Et3N
H(o)CRI4 NaCNBH3 Cl-heteroaryl ganic base IVx H~o~
where Y is _C~R8 or >C}NHR8 W095/306~2 -82- 634 r~ r~ lo ~ O
~O--~ Rs 7 (1) No2 R2 o R4 Identifiers/
NaBH4/~ ntifit~rc H2N(CH2)l 6R14 NaCNBH3, MeOH
b. \ NO
HS(CH2)2SH/
BF3-ether/
Identifiers \~
/(~` N~
NO2 R~ o R~
5a. 5b. 5c. 5d. 6 7 8 9 360 nm UV/alkanol t~ ' 21 ~634 WO 95/3064~ PCT/U59~/05940 BIS-LINKER ATTACHMENT
HO2C NHBoc DIC. HOBt. DMF
(~}NH2 + ~ ~
NHBoc (~N,I~, NHBoc H ~ 50% TFA,CH~CH2 NHBoc J~2 2 ~ P, 9 634 WOg5/306-12 F~l/-)~, ''i',1^ ~\
-8~

CLEAVABLE LINKER/Is LIGAND ELEMENT
(~NH2 + HO~O~< lQ
HOBt, DMF, DIC
ntifi~r~
O~
IV,~
I) TFA / DCM
2) NEt3 ~ DCM
o NO2 ~NHJ~,HN(CH2),~RI7 IV,~

2 ~ 8 q 6 3 4 WO95/30642 r~ s 1^

ATTACHMENT OF HYDROXYACETOPHENONES
.

H~2C~ ~OH

DIC, HOBt, DMF
IDENTIFIERS
IV, , ~ ~OH

r. 3 ~ 2~89634 W0 95/30642 F~ u~ 10 ADDITION OF R4/Rs ;~ R
0 5a' 5R~ R4 (X=CH2,O,S,N-Me) 5R~ R4 (X = N-t-Boc or CHNH(t-Boc)~
(~ N~ 17 R~ R~
Sc' 2 t 8 9 6 3 4 w Sc' CICO2R9 2,6 lutidine ~f.~f?nf,ifif.-rg/ OC 10 50%TFA/DCM NR
SCNR l I
r HO2R 12 ~d' (I) ~ DIC, DMF

Et3N
H~o)CRI4 NaCNBH3 Cl-hete~oa~
~ganic ba3e j V,~ R ~;~
where Y is ~R8 >O NHR
Me ~(CH 2)2C(O~ NR8 7 r~ 218 WO 95/30642 6 3 4 r~

ADDIl ION OF R6/R7 V ~ ~ Phe)o I ~ Rs T(l~nfifi~r~/NaBH4 Identifie~s/H2N(CH2)~ 6RI4 NaCNBH, MeOH
~ 3 \
H~,(CH2)l~6RI7 ¦
lVX NO2 ~(4-Phle)O I
T~lPnfifif~-c/ \ 0,~1 HS(CH~)2SH/\ R*=(CH2)~ 6RI4 ~ J~RS
BF3-ether \ ' 4 (~3 HN ~, NCH2) 1 6R17 8' (1) 2 o q~o ~_ R2; R4 Sa'. Sb'. Sc', Sd'. 6'. 7'. 8', 9' 360 nm UV/alkanol i~`; ` 7 ` ~ 2~ 89634 WO 95/30642 r~ ."10 -89- .

ADDITION OF Rl5

9.9' Reagents as in Scheme 9 (~3~ N~L~ NRIs(CH2)l 6RI4 l3 (1) NO2 R~ o R4 o P HN~,~CH2) l 6RI7 ~ 13' (I) ~Rs WO95130642 ` r~l".,~, ~tc, 1^

COMBINATOR~AL SYNTHONS
NO
O, NH2 + Ho~f (C~ Rl7 HOBt, DMF. DIC
o N2 0 (~}~ HJ~ (C~ 6RI7 I ) TFA / DCM
2) NEt3 / DCM

(~HR~HN(CH2), 6R17 Br ~I~ )`R =
X (Y)~l Br O I
o N2 )~ /` R
(~ H J~/N (Y~o~

X = OH, Cl, Br Y = aryl, heteroaryl R = H~ alkyl e~c 21 89634 WO95/30612 ` F~,l/~J~. '~'' I^
-91 - ~
.
Table 2 illustrates compounds of Formula 11 which are t;s~ dtive of the present invention:
~ Rs REPRESENTATIVE COMPOUNDS
lo Rl R2 R4 R5 R6 R7 6-OH 8-CH3 C2Hs C2Hs OH H

5-OH 7-CH2Hs H CH~Hs NH. H
6-0-(CH. )~OH H C3H7 CH3 =0 1~ 7-OCH2CO2H H -(CH2)4- H ¦ morpholino 8-O-(CH2)20H H -(CH2)5- N(CH3)2 ¦ H
6-C02H 8-CH3 -(CH2)6- -S(CH2)2S-6-OH H -(cH2)2o(cH2)2- =0 2C7-OH 8-CH3 CH3 ¦ CH3 -S(CH2~2S-6-OH H -(CH2)s- =0 - - ~ 2 1 8 9 6 3 4 WO95/30642 P~ ,. S.'~ 10 ~
-92- .
Table 3 illustrates ~d~ n~l compounds of Forrnula 11 scl-k Live of the present invention:

REPRESENTATIVE COMPOUNDS
RlR2 R3/R4 R6 R7 R8 6-OH H -(CH2)2NR8- OH H -CONH-Ph~-CF3 (CH2)2-

10 7-OH 8-CH3 -CH2NR8- '~ H -S02-2-Naph (CH2)3- 2 5-0(CH2)20H 7-c2H5 -(CH2)2NR8- _ N~ H -CSNH-Ph 6-OH H -(CH2)2NR8- =o -CO-Ph-4-S02NH2 (CH2)2-7-OH H -(CH2)2CH- -S(CH2)2S- -CO-Ph 1-so2NH2 (NR8)(CH2)2-2C~
6-OH H -(CH2)2NR8- =o -COCH2Ph (CH2)2-6-OH H -(CH2)2NR8- -S(CH2)2S- -C02-2-Py 257-OH 8-CH3 -(CH2)2NR8- -S(CH2)2S- -CO-Ph-4-S02NH2 (CH2)2-6-OH H -(CH2)2NR8- -S(CH2)2S- -CO-Ph-4-S02NH2 W095/30642 ~ .'i ,.f'~ 2 189634 r~l,u~ c~c ~o Table 3 (Corlt.) 7-OH H -(CH2)2NR8- =o -Co-Ph-4-S02NH2 (CH2)2-6-OH H -(cH2)2NR8- OH H CONH-Ph-4-C~3 (CH2).-7-OH 8-CH3 -(cH2)2NR8- N(CH3)2 H -S02-2-Naph 10 5-O(CH~)20H 7- -(CH2)2NR8- -SCH~CH- H -CSNH-Ph C2Hs CH2- (CH3!S-6-OH H -(CH2)2NR8- =o -CO-Ph-4-S02NiH2 (CH2)2-7-OH H -(CH2)2NR8- -S(CH2)2S- -CO-Ph-4-S02NH2 (CH2)2-6-OH H -(CH2)2NR8- =o COCH2Ph (CH2)2-6-OH H -(CH2)2NR8- -S(CH2)2S- -C02-2-Py 7-OH 8-CH3 -(CH2)2NR8- -S(CH2)2S- -CO-Ph ~ So2NH2 (CH2)2-256-OH H -(CH2)2NR8- -S(CH2)2S- -CO-Ph~S02NH2 7-OH H -(CH2)2NR8- =o -CO-Ph~S02NH2 (CH2)2-A '` ~ 21 ~ 9 6 3 4 WO 9~/306~2 P~ . 5.'~ ' 1^ ~

The invention is further defined by reference to the following examples, which are intended to be illustrative and not limiting.
PREPARATION
IDENTIFIERS
Twelve compounds of the general formula:
O ~ (CH2)n-O-Ar wherein:
n = 3-12 and Ar is pentachlorophenyl or n = 5-6 and Ar is 2,4,6-trichlorophenyl were prepared according to Scheme 13 and the following illustrative example.
a) Methyl vanillate (0.729 g, 4.0 mmol), 1-hydroxy-9-15 (2,3,4,5,6-pentachlorophenoxy)nonane (1.634 g, 4.0 mmol) and triphenylphosphine (1,258 g, 4.8 mmol) were dissolYed in 20 mL dry toluene under argon. DEAD (0.76 mL, 0.836 g, 4.8 mmol) was added dropwise and the mixture was stirred at 25'C for one hr. The solution was con~ontr~tPd to half volume and purified by flash c~uu-udlc~g-dphy 20 eluting with DC~ to give 1.0 g (1.7 mmol, 43%) of the product as a white crystalline solid.
b) The methyl ester from Step (a) (1.0 g, 1.7 mmol) was dissolved in 50 mL THF, 2 mL water was added, followed by LiOH
(1.2 g, 50 mmol). The mixture was stirred at 25 C for one hr. then 25 refluxed for 5 hr. After cooling to 25~C, the mixture was poured onto etnyl acetate (200 mL) and the solution was washed with I M HCI (3x 50 mL) then sat'd aq. NaCl (lx 50 mL) and dried over sodium sulfate.
The solvent was removed and the crude acid azeotroped once with toluene. The crude material was dissolved in 100 mL toluene, 10 mL
30 (1.63 g, 14 mmol ) thionyl chloride was added, and the mixture was refluxed for 90 min. The volume of the solution was reduced to p ,~ t ~ 2 1 8 9 6 3 4 WO 951306~2 F~ .'u' I^
_95_ approx. 30 mL by ~ till~ti~m, then the remaining toluene was removed by evapordtion.
c) The crude acid chloride from Step (b) was dissolved in 20 mL dry DCM and cooled to -70C under argon and a solution of S approx. 10 mmol dia~u~ Ll~dl~e in 50 mL anhydrous ether was added.
The mixture was warmed to r.t. and stirred for 90 min. Argon was bubbled through the solution for 10 min., then the solYents were removed by evapordtion and the crude material was purified by flash cl~ .atoglal,lly, eluting with IQ-20% ethyl acetdte in hexane. The 10 diazoketone (0.85 g, 1.4 mmol, 82% yield over three steps) was obtained as a pale yellow solid.
In alternate Step (c) there is a change to the final diazo-methylation step, whereby the â~id chloride is reacted with (trimethyl-15 silyl)did~ulll~llldile and triethylamine to give the identifier, which canthen be used without further puri~lcation. With this alternate step, the identifier can be obtained in high yield with no chloromethylketone byproduct. Also, purification by flash chromatogrdphy is no longer necessary, which in some cases h~s resulted in ~i~nifir:~nt acid-catalyzed 20 de~olllposi~ion of the identifier.
Alternate Step c). To a solution of the acid chloride (3.8 mmol, 1.00 equiv.) and 1.85 mL (13.3 mmol, 3.50 e~uiv.) of triethylamine in anhydrous THF/~cetonitrile (1:1) at O C under argon was added 5.7 mL (11.4 mmol, 3.00 equiv.) of â 2.0 M solution of 25 (trimethylsilyl)~ 7~m~h~n~ in hexanes. Th~ resulting ordnge solution was stirred at O C for 2 hr, then at 25 C for 17 hr. (If a pl~ a formed imml~di~ ly upon addition of (trimethylsilyl)diazomethane, CH2CI2 was added until the preCipitdte redissolved). EtOAc was added (250 mL), and the organic layer washed with saturdted aq. NaHCO3 30 (100 mL) and H20 (IOQ mL), then dried (anhydrous MgSO4). Removal of the volatiles in vacuo gave the product as yellow crystdls in 60-100%
yield.
The other 11 i~ nfifif-rs of Formula IV were prepared by analogous synthetic routes, steps (a), (b), and (c).

r~ rr~ 2 1 8 9 6 3 4 WO 95/30642 F~,ll~J,.,''~,Jg ~0 ln the synthesis of Example 1, the 12 identifiers were used to encode the combinatorial library. In Step 1, pentachlorophenyl i~lPlltifiPT~, where n = 11-12 (abbreviated Cl lCls and CI~Cls were used in the following binary encoding scheme: Ol = (n = 12) and 10 = (n =
S 11). In Step 2, pentachlorophenyl i~ntifiPrs where n = 8-10 (abbreviated CgCls, CgCls, and C1oCls) were used and encoded as follows: OOI = (n = 10), O10 = (n = 9), and 100 = (n = 8). In Step 3, pentachlorophenyl i~lPntifiPr~ where n = 3-7 (abl,l~vial~d C3Cls, C4Cls, CsCIs, C6Cls, and C7Cls) were used and encoded as follows:
10 00001 = (n = 7), O~O10 = (n = 6), OO100 = (n = 5), O1000 = (n = 4), and 10000 - (n = 3). In Step 4, trichlorophenyl i~lPn,~ifiPr.~i where n =
5-6 (al~l,lcv~ d CsC13 and C6C13) were used and encoded as follows:
Ol=(n=6) andlO=(n=5).
Thus, in Step I reagent 3 (Table 1-1) is encoded " I l "
15 which ~ ,s~llL, tagging this choice in the synthesis with the two pentachloro-phenyl i~1PntifiPr.C where n = 11 and 12.

~ t~ 2 ~ 8 9 6 3 4 WO 95/30612 , IDENnFlERS
OH HO-(CH2)n-O-Ar MeO~bMe PPh3, DEAD.Toluene ~~(CH2)n~0~Ar MeO~`OMe 1. LiOH, THF/MeOH
2. SOC12, toluene reflux Cl~o--(CH2)n-O-Ar TMS - CHN2 OMe Et3N /
/C. THF/MeCN (I:l) CDHc2MN2 Et20 N~O--(CH2)n-O-Ar O OMe lV

è r ~ 21 89634 WO95/30612 F~

t-BUTYL 4-(HYDROXYMETHYL)-3-NITRO~ENZOATE
t-Butyl 4-(acetoxymethyl)-3-nitrobenzoate was prepared as described by Barany and Albericio, J. Am. Chem. Soc. 1985, 107, 49364942. The reference's final ~lucedul~ for hydrazinolysis of the acetate using hydrazine hydrate in CHC13 at 25 C produces only trace amounts of the desired hydroxymethyl final product, which is the t-butyl ester pre-cursor of the photocleavable linker used herein.
However, hydrazinolysis using hydrazine hydrate in MeOH at 25 C
produces t-butyl 4-(hydroxymethyl)-3-nitrobenzoate in high yield.
Using MeOH as solvent, only the desired final product is obtained in near 4ua~ yield (93%).
t-Butyl 4-(hydroxymethyl)-3-nitrob~n70~t~: To a solution of 14.1 g (47.7 mmol, 1.00 equiv.) of t-butyl 4-(acetoxymethyl)-3-nitrobenzoate in MeOH (200 mL) was added 27.0 mL (477 mmol, 10.0 equiv.) of hydrazine hydrate (55% hydrazine). The resulting yellow solution was stirred at 25'C for 4 hr. EtOAc (250 mL) and sâturated aq. NâCI (85 mL) were added, and the organic layer collected after shaking. The orgânic layer was washed further with saturated aq. NaCI (2 x 85 mL), and then dried (MgSO4). Removal of volatiles in vacuo gave the product in g3% yield as yellow crystals.

ALLYL 4-(HYI:)ROXYMETHYL)-3-NITROBENZOATE
In a 100 mL round bottom flask was placed ~hydroxymethyl-3-1li~lubtl~oic acid (1.97 g, 10 mmol). Allyl alcohol (20 mL) was added, followed by p-toluenesulfonic acid (0.190 g. 1 mmol). The mixture was heated to reflux for 24 hr., at which time all the volatiles 30 were removed in vacuo. The residue was taken up in EtOAc and washed with sat'd KHCO3. The organic layer was dried over MgSO4 and cu" ~ ,.t~d to âfford the title compound as a cream colored solid;
2.4 g (100%).

W0 951306,~2 I ~ I / U J~ C~

METHYL 4-(HYDROXYMETHYL)-3-NITROBENZOATE
Following the procedure of Preparation 3, but using methanol instead of allyl alcohol, the title compound was prepared in 5 57% yield.

BIS-LINKER MQDIFIED RESIN
Step 1 Addition of bis-Boc Iysine In a 250 mL synthesis vessel was placed bis-Boc-(L)-lysine 10 (7.71g, 22.2 mmol) as a solution in DMF (150 mL). HOBt (2.84g, 21.0 mmol) was added followed by DIC (3.25 mL, 21.0 mmol) and the solution shaken at r.t for 15 min. before adding TentaGel resin (25.8 g, appr~ximately 7.2 mmol amino sites). The mixture was shaken at r.t.
for 17 hr and then washed alternately with methanol and DCM (SX
15 each) and then with THF (2X) and dried under vacuum.
Step 2 Deprotection Into each of seven 250 mL synthesis vessel was placed modified TentaGel resin (8.0 g, approx. 4.5 mmol of N-Boc amine sites). DCM (75 mL) was added followed by a 30 % TFA solution in 20 DCM (100 mL). The vessel was shaken at room ~ p~,ldlul~ for 15 min before adding neat TFA (15 mL). The vessel was shaken at room f ."ll.,G for 2.5 hr at which time the resin was washed with DCM
(2X). The resin was then treated with a solution of 10% t~iethylamine in DCM (2X150 mL) shaking for 20 min. each time. The resin was 25 then washed with DCM (4X) and DMF (lX).

t-Boc-PROTECTED AMINO ACID
In a I L flask was placed 3-nitro-4-(bromomethyl) benzoic acid (20.03 g, 77.0 mmol). THF (300 mL) was added followed by 4-30 methoxyl,~.~yla,~ e (10.0 mL, 77.0 mmol) and triethylamine (35 mL).The resulting clear solution was stirred at r.t. for 17.5 hr. Solid di-tert-butyl dicarbonate (16.8 g, 77.0 mmol) was added, followed by DMF (100 mL) and the resulting suspension stirred at r.t. for 72 hr.
_, _ _ _ .. . .... ... .... .. ..... ..... .. . . . .... . . ...

'tS 218~634 The reaction mixture was (~onc~ntr~f~d in vacuo and the residue taken up in ethyl acetate, washed with I N HCI (X2), dried (Na2SO4), filtered and cu..c~llt-dled to afford a dark bro~vn oil. Purification via flash cl..ullldlu~laphy (ethyl ~eet~t.?:h~xane) resulted in a yellow foam which 5 was triturated with acetonitrile to give the expected protected amino acid (Table 2-1, compound 4) as a fine white powder (9.9lg, 31%).

t-Bûc-pR~TEcTFl~ AMI~ A~rn Sllhst~nti~lly fûllowing the plucclul~ ûf ~ al~tiOII 6, but ~ ;l.g the ~ luL,-idle amine for 4-methoxybenzylamine, the aillillg cûlll~ùullds of Table 2-1 are prepared.
E~XAMPLE 1 1299 COMPOUND l lBRARY
15 Step 1 a) RI/R2 To a solution of t-butyl 4-hydroxymethyl-3-nillubel~udl~
(2 g, 7.89 mmol, I equiv.), 2,~dihydroxyacetophenone (1.20 g, 7.89 mmol, I equiv.), and triphellyll~h~ hin~ (2.69 g~ 10.26 mmol, 1.3 20 equiv.) in toluene (20 mL) was added dropwise DEAD (1.79 g, 10.26 mmol, 1.3 equiv.). After addition was complete the mixture was stirred for 16 hours at room ~ lalul~. The solvent was removed in vacuo and the residue was purified by flash chromatography (SiO2, eluted with 10% ethyl acetate in hexanes) affording 1.47 g ûf the product 25 (48% yield).
The t-butyl ester (500 mg, 1.29 mmol, I equiv.) above was dissolved/suspended in DCM (8 mL) and tre~ted with TFA (3 mL).
The mixture was stirred at room temperature for 8 hours. The DCM
and TFA were removed in vacuo affording a white sûlid. This was 30 a~cutluped once with toluene then dried in vacuo affording 427 mg (100% yield) of the carboxylic acid.
The acid (636 mg, 1.92 mmol, 1.5 equiv.) prepared above was dissolved in DMF (40 mL) and added to divinylbenzene-cross-linked, polyethyleneglycol-grafted polystyrene beads (TentaGel(~) S

?~T~. 2189~34 NH2, Rapp Polymere) (4.0 g, 0.32 mmol/g, 1.28 mmol, 1 equiv.) in a Merrifield reaction vessel. The resin was sllcpçn-3~d by agitation, then HOBt (259 mg, 1.92 mmol, 1.5 equiv.) and DIC (0.31 mL, 1.92 mmol, 1.5 equiv.) were added in that order. The resin was agitated at room 5 L~ tldlul~ for 7 hours at which time it gave a negative Kaiser test.
The resin was filtered and washed (DMF 3x50 mL, DCM 3x50 mL) then dried in v~.~cuo.
The two other dihydroxyac~ l~hel~ s were attached to the resin via the photocleavable linker in an analogous manner using the 10 reagents of Table 1-1.
Alternate a) In an analogous fashion the allyl and methyl esters were prepared from allyl 4-hydroxymethyl-3-nitrobpn7o~ (Preparation 3) and methyl 4-hydroxymethyl-3-1li~lu~l~odL~ (Preparation 4).
In a 10 mL flask was placed the allyl ester (110 mg. 0.3 mmol). Methylene chloride (2 mL) was added followed by tetrakis-triphenylphospine F~ lm(O) (11.5 mg, 0.01 mmol) and the mixture cooled to 0'C. Pyrrolidine (50 mL, 0.6 mmol) was added and the reaction stirred at 0C for 45 min. The mi~ture was diluted with 20 EtOAc (10 mL) and washed with 3.5N HCI. The organic layer was dried (MgSO4), filtered, and concentrated to afford a yellow solid; 90.6 mg.
In an analogous mamner the methyl ester was deprotected by basic hydrolysis using a mixture of dilute NaOH and THF.
25 b) Encoding of Step I
Quantities of the three resin batches (2.5 g) from Step l(a) were placed in separate synthesis vessels and each was s~lcren~ d in DCM (20 mL). The three appropriate binary coding mixtures (Table 1-1) for each batch of resin were prepared by dissolving the 30 apL~Iu~lid~c choice (37.5 mg) or choices (37.5 mg of each) of Cl2CI5 and Cl~Cls -linker diazoketone (Preparation 1) in DCM (1 mL for each solution). These solutions were added to the d~lUI)lid~t~ synthesis vessel and the resin was agitdted for 30 mins.
Rhodium trifluoroacetate dimer (I mL of a I mg/mL
35 solution in DCM) was added to each of the vessels and the resin was -r~ t ~ 2 1 89634 WO95/30612 r~

agitated at room telll,u~latul~ for 4 hours. Each batch of resin was then filtered and washed with DCM (2x20 mL) then each was rP~I~cpPnrlPd in DCM (20 mL) and treated a second time with the ap~uplial~ binary encoding mixture as described above. The resin was again agitated for 5 30 mins before addition of the rhodium triflouroacetate dimer. The same quantities of catalyst and ~ 7okpt~np cu~ Jvuud~ were used in the second coupling step as in the first. The resin was agitated for 14 hours. Each resin batch was then washed with DCM (5x20 mL) then the batches were combined and the entire library (three compounds) 10 was washed with DCM (l0x50 mL).
Ste~ 2.
a) Cycli~c~ P~ n Reactions The dried resin from Step I O was divided into four batches of 1.5 g (ca. 0.42 mmol) and three ~1rliti~ batches of 0.2g 15 (~. 0.056 mmol). The 1.5g batches were placed into 25 mL round-bottomed flasks and the 0.2 g batches were placed into 5 mL round-bottomed flasks. The portions of resin were sl-cpPnrlPd in methanol (15 mL in the four flasks with 1.5 g of resin, 2 mL in the three flasks with 0.2 g of resin) and pyrrolidine (0.6 mL, 7.2 mmol, ~. 15 equiv. in the 20 flasks with 1.5 g of the resin; 0.08 mL, 0.96 mmol, ~. 15 equiv. in the flasks with 0.2 g of resin) was added to each flask. The reaction vessels were then allowed to stand for 5 min. to allow mixing of the reagents.
The d~ ulJIial~ ketone (>10 equiv.) was then added to the vessels. The four BOC protected ~min-)kPt~-n~ were added to the flasks containing 25 1.5 g of resin and the other ketones were added to the flasks containing the 0.2 g of resin. The mixtures were heated at 75C for 16 hr. The flasks were then cooled to room t~ dLul~ and each batch of resin was poured into a separate sintered funnel and washed thoroughly with DMF (3x20 mL) and DCM (3x20 mL).
b) Encoding of Step 2 Each batch of resin from Step 2(a) was placed into a separate synthesis vessel and was suspended in DCM (5 mL for the batches of 1.5 g of resin, I mL for the batches containing 0.2 g of 35 resin). The seven d~.lu~l;al~ binary coding mixtures (see Table 1-2) for each batch of resin were prepared by dissolving the appropriate ç ~ 9 6 3 4 ~ WO 95130642 F~ J,,,''C', ~^

choice (22.5 mg if added to a batch of 1.5 g of resin; 3.0 mg if added to a batch of 0.2 g of resin) or choices (22.5 mg of each if added to a batch of 1.5 g of resin; 3.0 mg of each if added to a batch of 0.2 g of resin) of Clocls~ C9C15, and C8Cls linker-diazoketone (Preparation I ) in DCM (I mL for each solution). These solutions were added to the al~,u~uplidl~ synthesis vessel and the resin was agitated for 30 mins.
Rhodium trifluoroacetate dimer (I mL of a 1 mg/mL
solution in DCM) was added to each of the vessels and the resin was agitated at room ~ ld~U~C for 4 hr. Each batch of resin was then filtered and washed with DCM (2x20 mL) then each was rPcllcpPn~Pd in DCM (5 mL for the batches of 1.5 g of resin, 1 mL for the batches of 0.2 g of resin) and treated a second time with the d~J~lU~Jlid~t; binary encoding mixture as described above. The resin was again agitdted for thirty mins before addition of the rhodium trifluoroacetate dimer. (The same quantities of catalyst and did~ olle compounds were used in the second coupling step as in the first.) The resin was then agitated for 16 hr. Each resin batch was then washed with DCM (Sx20 mL). The four batches of 1.5 g of resin were combined and washed with DCM (lOxS0 mL). These combined batches were then reacted further in Step 3.
The three batches of 0.2 g of resin were combined and washed with DCM (lOx20 mL). These combined batches were not used in Step 3 but were saved for Step 4.
Step 3 a) Encoding of Step 3 25. The four batches of 1.5 g of resin which had been combined in Step 2(b) were now divided into thirty lots of 170 mg each in I dram shell vials (Fisher Scientific) and each was suspended in DCM
. (2 mL). The thirty dpl)lul~lidl~ binary coding mixtures (see Table 1-3) for each batch of resin were prepared by dissolving the a~J~lu~Ulidl~
choice (3 mg) or choices (3 mg of each) of C7C15, C6cl5~ CsCIs, C4 and C3C4 linker-diazoketone (Preparation 1) in DCM (I mL for each solution). These solutions were added to the dpl lu~lidL~ synthesis vessel and the resin was agitated for 30 mins.
Rhodium trifluoroacetate dimer (I mL of a I mg/mL
solution in DCM) was added to each of the vessels and the resin was agitated at room l~ d~ul~ for 4 hr. The supernatant solution was c ~ ~ 8 ~ 6 3 4 WO 95/306~12 ~ 10 ~

then decanted away from the resin with a Pasteur pipette. The resin was washed twice with DCM (3 mL) and the washings removed by Pasteur pipette. Each batch of resin was resuspended in DCM (2 mLl and treated a second time with the appropriate binary encoding mixture 5 as described above. The resin was again agitated for thirty minutes before addition of the rhodium trifluoroacetate dimer. (The same quantities of catalyst and ~ k~t~n~ compounds were used in the second coupling step as in the first.) The batches of resin were then agitated for 16 hr. Each resin batch was then l.dllar~.,,,d to a small 10 Merrifield reaction vessel and washed with DCM (3xlS mL), DMF
(2xlS mL), and DCM again (2x15 mL).
b) D~plvL~iol~
Each batch of resin was treated with a 50% solution of 15 TFA in DCM (6 mL:6 mL). The resin was agitated for 2 hr and then filtered and washed with DCM (3x15 mL). The resin was then treated with a 10% solution of triethylamine in DCM (I mL:9 mL) and agitated for 10 mins. This treadtment was repeated once. The resin was filtered and washed with DCM (4xlO mL).
c) Addition of R~
To each of the first six flasks was added DCM (5 mL) and the resin was agitated for 10 mins. 2,6-Lutidine (0.11 mL, 20 equiv.) was added to each flask followed by a solution of the a~L~Ivplid~e 25 chlvlorv-lllat~ (Table 1-3) in DCM (S mL) and the resin was agitdted for 4 hr. Except for isopropylchlolvrullllàte (Aldrich), the chlvlurvlllld~,a were prepared from the dp~JIuplid~ alcohols by treating the alcohols (0.1 g) with a solution of phosgene in toluene (5 mL of a 1.8 M solution) for 1 hr, then evaporating to dryness in vac~lo 30 ,and then redissolving in DCM (5 mL).
To flasks 8, 9, and 10 was added ethanol (10 mL) and the apl,lvplia~ isocyanate (Table 1-3) (0.1 mLI ca. 10 equiv.) and the resin was agitated for 4 hr.
To flasks 11, 1, and 13 was added ethanol (10 mL) and 35 the d~ u~Jlidl~; isothiocyanate (0.1 mL, 0.1 g of the n~rhth~l~n~
isothiocyanate, ca. 10 equiv.) and the resin was agitated for 4 hr.

WO 9~/30642 F~,l/lJ,.,~'~,5, I^

To flasks 7 and 14-22 was added DMF (10 mL) and the d~plU~lidt~ carboxylic acid (ca. 10 equiv.) and HOBt (0.103 g, ~. 15 equiv.). The flasks were agitated for 30 mins then DIC (0.12 mL, ca.
15 equiv.) was added to each flask and the resin was agitated for 4 hr.
S To flasks 23-30 was added DCM (10 mL) and triethylamine (0.15 mL, ca 15 equiv.) and the resin was agitated for 15 mins. The dL~lU~lial~ sulfonyl chloride (ca. 10 equiv.) was added to the reaction vessels and the resin agitated for 4 hr.
The flasks were filtered and the resin washed with DCM
(3xl0 mL). All of the resin was combined in one large synthesis vessel and was washed with DCM (3x50 mL), DMF (3xS0 mL), and DCM
again (3xS0 mL). The resin was dried in vac~o.
Altemative Step 3 a) Encoding of Alternative Step 3 . The remaining 900 mg of resin from the four combined batches of 1.5 g from Step 2(b) which had not been used in Step 3 was divided into ten portions of 90 mg. and each portion placed in a separate 1 dram shell vial (Fisher Scientific). The ten a,u~lu~,lid~
binary coding mixtures (see Table 1-5) for each batch of resin were prepared by dissolving the ap~lu~,idle choice (1.5 mg) or choices (1.5 mg each) of C7Cls, CsCls, C4Cls, and C3Cls linker-did~ùk~lc lle (~Ic~d~ ion 1) in DCM (1 mL for each solution). These solutions were added to the d~lu,u-idl~ synthesis vessels and the resin was agitated for 30 min.
Rhodium llinu~luac~ dimer (1 mL of a 1 mglmL
solution in DCM) was added to each of the vessels and the resin was agitated at room l~ ueldtul~ for 4 hr. The aulJ~IIld~dllL solution was . then decanr~ed from the resin. The resin was washed (DCM 2x 3 mL) and the washings removed by Pasteur pipette. The resin was then treated a second time with solutions of the d~l~luplia~e binary coding mixtures and agitated for 30 min. before the addition of the rhodium trifluoroacetate dimer. The same quantities of catalyst and diazoketone compounds were used in the second coupling as in the first. The - batches of resin were then agitated for 16 hr. The resin was then I~dn~r~ d into small Merrifield synthesis vessels and washed (DCM 6x 15 mL).

P5~ Sl ~1 89~34 WO95/306~2 F.l/~J,.,3,`~, 10 b) Deprotection.
Each batch of resin was treated with a solution of TFA in DCM (4 mL:4 mL). The resin was agitated for I hr, then filtered and washed with DCM (2x 15 mL) The resin was then treated with a 5 solution of piperidine in DCM ~4 mL:4 mL) and agitated for 15 min.
This treatment was repeated once. Each batch of beads was washed with methanol (2x 15 mL) and ~CM (4x 15 mL). Flasks 1-4 were washed with THF (3x 15 mL).
c) Heteroarylation Reactions The resin in flasks 1~ was s--cr~ n~ d in THF (6 mL).
Flasks 1-3 were then treated Wi'~l DBU (190 IlL, ca 40 equiv.) followed by the dlJIUlU~lia~ h~feroaryl ~hloride (ca 20 equiv.). Flasks 1 and 2 w~re heated at 55 C f~r 16 hr. Flask 3 was heated at reflux for 16 hr. Flask 4 was treated v~ith tri~llyla."i"e (700 ,uL) and the 15 app.opriate heteroaryl chloride (ca 20 equiv.). The resin was shaken at r.t. for 16 hr. Each batch of resin was then washed in THF (2x 15 mL) and dried in vacuo.
d) Reductive Alkylations The resin in flasks 5-10 was sl~rçn~d in DMF (8 mL) and 20 the d~lu,up.iale aldehyde (ca 67 equiv.) added. Acetic acid (160 ~aL) was added to each of the flasks t`ollowed by sodium cyanoborohydride (ca 67 equiv.). Flasks 5, 6, 7, 9~ and 10 were shaken at r.t. for 16 hr.
Flask 8 was heated to 55 C for 16 hr. Each batch of resin was filtered and washed with DMF (3x 15 m~). Each of the reductive alkylation 25 reactions was repeated under the same conditions. The batches of resin were washed with DMF (2x 15 rlL), methanol (3x 15 mL), and DCM
(3x 15 mL). The resin was then mixed, washed with DCM (2x 20 mL), and dried in vacuo.
This part of the library did not undergo further 30 elaboration.
Step 4 a) Encoding of Step 4 To the combined resin from Step 3(c) was added 45 mg of resin from each of the seven flasks from Step 2(b) and the resin was 35 washed and mixed thoroughly with DCM (3xS0 mL). From this ?~s~Q~ 21 89634 WO9~130542 r~ L",~ c 1^

mixture was weighed out three portions of 800 mg of resin and these were placed into three separate Merrifield synthesis vessels and suspended in DCM (10 mL). The three d~u~luplidl~ binary coding mixtures (see Table 1-4) for each batch of resin were prepared by S dissolving the a~u,ulvplidL~ choice (24 mg) or choices (24 mg of each) of the CkCI3 and C5C13 linker-diazoketone compound in DCM (I mL for each solution). These solutions were added to the dL~uluplia~ synthesis vessel and the resin was agitated for 30 mins.
Rhodium trifluoroacetate dimer (I mL of a I mg/mL
solution in DCM) was added to each of the vessels and the resin was agitated at room ~ dlUI~ for 4 hr. Each batch of resin was then filtered and washed with DCM (2x20 mL) then each was l~ ,e~h~d in DCM (10 mL) and treated a second time with the dL,uluplidle binary encoding mixture as described above. The resin was again agitated for 30 mins before addition of the rhodium trifluoroacetdte dimer. The same quantities of catalyst and ~ 7n~fon~ compounds were used in the second coupling step as in the first. The resin was agitated for 14 hr.
Each resin batch was then washed with DCM (3x20 mL) and then filtered.
b) Carbonyl Reaction (addition of R6 and R7) The resin in flask I was resuspended in DCM (6 mL) and 1,2-eth~n~-~ithiol (l mL) and boron trifluoride etherdte (I mL) were added. The flask was shaken at room ~elllp~-dlul~ for 6 hr. The resin was then washed with DCM (20 mL) and then r~sl-sr~nd~d in DCM (6 mL) and treated once more with eth~ln~difhi~ l (I mL) and boron trifluoride etherate (I mL). The resin was agitated at room t~ dlUle for 14 hr. The resin was then filtered and washed with DCM (Sx2.0 mL).
The resin in flask 2 was suspended in methanol (5 mL) and solid sodium borohydride (200 mg) was added cautiously. The flask was vented and allowed to shake gently for I hr. The resin was filtered and r~c~lcren~l~d in methanol and the reduction process repeated a tOt of 5 times at I hr intervals using 200 mg of sodium borohydride each time. After the final cycle the resin was washed with methanol (3x20 mL) and DCM (3x20 mL).
The resin in flask 3 was not reacted further.

c 21 8963~
WO 95/306~2 P~ ~ 0. ~

The resin from the three flasks was combined and washed with DCM (5x50 mL) and then dried in ~acuo. A portion (500 mg) of the resin was suspended in DCM (5 mL) and TFA (5 mL) and shaken for 2 hr. The resin was then treated twice with a 10% solution of 5 triethylamine in DCM (10 mL) and washed with DCM (Sx20 mL). The resin was then dried in vacuo.
d) Decoding Procedure A bead was placed in a 1.3 mm diameter pyrex capillary with 2 ~IL of acetonitrile. Ceric ~mmnnillm nitrate solution ~2 IIL of a 10 0.1 M aq.. solution) and hexarle (3 IIL) were added and the two-phase mixture c~ llL.irul5~d briefly. The tube was sealed and left at 35 C for 16 hrs, then opened. The organic layer was removed by syringe and mixed with I IIL of N,O-bis(trimethylsilyl)~ce~mi~i~ The silated tag solution (I ,uL) was analyzed by GC with electron capture (EC) 15 detection.
The GC analysis was performed with a Hewlett Packard 5890 plus gas cLv~ v~ On column injection into a 5 m, 0.32 mm retention gap connected to a 25 m, 0.2 mm crosslinked 5%
p~ .lyllll~Lllyl silicone column was used. The l~;lllp~ Ult~ program was 20 set at 200 C for I min and then increased at a rate of 15-C/min from 200-320 C. The pressure program was set at 20 psi for I min, then increased at 2 psi/min to 40 psi with a total run time of 10 min. The EC detector was l" .i"l~i.,fd at 400 C and the auxiliary gas was set at 35 psi.

2~ 8963~ -9~/30642 ~ C 1^
-109- ' ' Table 1-1 _1/R2 Rea~ents and Encodin~ S~hPm~ .
~ ~ ~o 1. 01 2. 10 ~O~OH
HO~ Me 3. 11 O
Table 1-2 R~5 R~a~ents and Encodin~ Scheme ~ BOC ~N~ BOC NH ~
1. 001 2. 010 4. 100 3. 011 5~1 6. 110 7. 111 woss/30642 h ~ / C 2 J 8 96 34 ~""~

Table 1-3 R~ Rea~ents and Encodin~ Scheme o~5~C~ CH2CHCH2è~, 5 1. 00001 2. 00010 3. 00011 <~~5~C~ 0~C~ rO'~CI
4. 00100 ~. 00101 6. 00110 NC~CO2H ~N=C=O Cl~tl=C=O
7. 00111 8. 01000 9. 01001 F3C~:=C=O ~ ~C~S ~3N=C--S
0. 01010 11. 01011 12. 01100 N=C=S - o H O
2(1 ~ ~COO2H O~CO2H
13. 01101 14. 01110 16. 01111 6~Co2H ~H N~C02H
16. 10000 17. 10001 18. 10010 25 . ~f CO2H ~ ~CO2H
- 19. 10011 20. 10100 21. 1010 H2NO2S J~C02H ~2CI SO2C
22. 10110 23. 10111 21.11000 ~1 89634 C
WO 95/30642 `'~ r~ ,05$1 CI~SO2CI Me~S02CI G~so2c 25. 1 IOOl 26. l l010 27. l l0] 1 ------SO2CI >--S02CI Cl02S~t-Bu 28. l1100 29. 11101 30. 11110 Table 1-4 _~/Rl Rea~ents lml En~otlin~ S-~h~ m~
¦ HS(CH2)2SH/BF3-0Et2 ¦ NaBH4 ¦ No reaction 1.01 12. 10 13. 11 Table 1-5 R8 and Heteroaryl Encodin~ Scheme >--Cl ~ ~NH2 1. 0001 2cl 0010 2~ N~NCI cl~
3. 0011 4 0100 F~CHO H3CO~CHO
5. 0101 6. 0110 ~CHO N~CHO
7. 1000 8. 1100 ~--CHO ~CHO
9. 1010 10. 1001 87.906 COMP()UND LIBRARY
Stçp I
a) Addition of (CH2) 1 6R17 S In a 2~0 mL synthesis vessel was placed the deprotected modified TentaGel resin (8.0 g, approx. 4.5 mmol of amine sites) from Preparation 5. HOBt (1.81g, 13.4 mmol) was added followed by the N-Boc-p-methoxybenzylamino acid (Table 2-1, compound 4) (5.60g, 13.4 mmol) and DMF (150 mL). The mixture was shaken at r.t. for 10 min.
before adding DIC (2.1 mL, 13.4 mmol). The mixture was shaken at r.t. for 16 hr and then washed alternately with methanol and DCM (4X
each) and then with EtOAc (2X). Analysis of the resin via the standard Kaiser ninhydrin test indicated that the coupling reaction was complete.
In six separate vessels, analogous couplings were carried out with the six other Boc-protected amino acids listed in Table 2-1.
All coupling reactions were repeated until ~ r~ Oly Kaiser ninhydrin test results were obtained (in all cases either one or two couplings).
b) Encoding of Step I
While still in their separate 250 mL synthesis vessels, resin batches number 4, 5, 6, and 7 from Step I were suspended in EtOAc (100 mL). Into each of these four vessels was placed the ClsC7-linker dia~oketone (0.56 g) and the mixtures agitated for I hr. To each of the four vessels was then added rhodium L~illuuludc~ldte dimer (6 mL of a I mg/mL solution in DCM) and the resin was agitated for 15 hr. The resin was then washed with DCM (4X) and EtOAc (2X).
In turn, the ClsC8-linker diazoketone was applied to resin batches numbered 2, 3, 6, and 7 and the ClsCI l-linker tli~70k~tt~Dc was applied to resin batches 1, 3, 5, and 7. Application of each tagging molecule was done separately and in analogous fashion to that of the ClsC7-linker ~I;A~UI~rI~ O outlined above. The seven batches of encoded resin were all combined in a 21. Erlenmeyer flask along with THF (I
L) and mixed thoroughly by swirling and stirring gently with a glass rod. The resin was then recovered by filtration and vacuum dried.

2 ~ ~9634 ~ WO 95/30642 r~ u.. 35'~

Step 2: Addition of R2 a) Deprotection In a 250 mL synthe~is vessel is placed mixed, encoded resin from Step I (9 g) along with DCM (just enough to suspend resin). TFA
5 (75 mL of a 30 % solution in D('M) is added and the resin agitated for 3.5 hr. The resin is then wash~d with DCM (2X) followed by treatment with 10% triethylamine in DC~ (2X 20 min. each) and then washed with DCM (4X).
b) Coupling The deprotected re~in from Step 2(a) (9 g) is suspended in DMF (7 mL). HOBt (2.04 g, 1~ inmol) is added followed by the acetophenone acid (Table 2-2, compound 31 (3.36g, 15 mmol) and the mixture agitated for 15 min. D~ (2.3 mL, lS mmol) is added and the mixture agitated for 21 hr. The xsin is washed alternately with DCM
15 and methanol (SX each) and then with EtOAc (4X).
In five separate vessels, analogous couplings are carried out with the five other acetophenone acids listed in Table 2-2.
c) Encoding of Step 2 The six batches of resin from Step 2 are binarily encoded 20 in a fashion analogous to that described above for encoding of Step I
The six batches of encoded resin are combined in a 2 L
Erlenmeyer flask along with TH~ (I L) and mixed thoroughly by swirling and stirring gently with a glass rod. The resin is then recovered by filtration and vacuum dried.
25 Step 3 Addition of R4R5 a) Cyclocon~lenc~tion reactions The mixed resin from Step 2 is divided into three batches of 14.4 g (ca 8.1 mmol) and seven ~d~liti~n~l batches of 1.5 g (ca 0.84 mmol). The 14.4 g batches are placed into 250 mL round bottom flasks 30 and the 1.5 g batches are placed in 25 mL round bottom flasks. The portions of resin are suspended in methanol (150 mL in the three flasks with 14.4 g resin, 15 mL in the seven flasks with 1.5 g resin) and pyrrolidine (10.1 mL, 121 mmol, ca 15 equiv. in the flasks with 14.4 g resin; 1.0 mL, 12.6 mmol, ca 15 equiv. in the flasks with 1.5 g resin) is ~, ~ `, fl,~ ~ C
W0 95/306~2 2 1 8 9 6 3 4 F~

added to each flask. The reaction vessels are then allowed to stand for 15 min. to allow mixing of the reagents. The d~lVpl;al~ ketone (5 to 10 equiv.) is then added to the vessels. The three Boc-protected ~",i"n~ c from Table 2-3 are added to the flasks containing 14.4 g 5 of resin and the seven other ketones, from Table 24, are added to the flasks cl,,ll~i,,i,,~ 1.5 g of resin. The mixtures are heated at 75C for 16 hr. The flasks are then cooled to r.t. and each batch of resin is poured into a separate synthesis vessel of ~ ul~iiaie size and washed thoroughly with DCM, DMF, and methanol (~Itf~rn:~tin~: SX each).

b) Encoding of Step 3 Each of the ten batches of resin from Step 3(a) is binarily encoded in a fashion analogous to that described for encoding Step 1.
c) Mixing and dividing The seven 1.5 g batches of encoded resin are combined in a 500 mL Erlenmeyer flask along with THF ~250 mL) and mixed thoroughly by swirling and stirring gently with a glass rod. The resin is then recovered by filtration and vacuum dried. This combined resin is kept separate from the three 14.4 g batches of resin and is not 20 subjected to the reaction conditions of Step 4, but rather re-divided into three 0.2 g portions and seven 1.4 g portions and saved to be used in Step 5 and alternate Step 5. The three 14.4 g batches of encoded resin are combined in a 2 L Erlenmeyer flask along with THF (I L) and mixed thoroughly by swirling and stirring gently with a glass rod. The 25 resin is then recovered by filtration, vacuum dried, and used in Step 4.
Step 4 a) D~lu~lioll Into each of seven 250 mL synthesis vessels is placed mixed, encoded resin from the three combined 14.4 g batches from Step 30 3 (6 g) along with DCM (just enough to suspend the resin). TFA (75 mL of a 30 % solution in DCM) is added and the resin agitated for 3.5 hr. The resin is then washed with DCM (2X) followed by treatment with 10% triethylamine in DCM (2X 20 min. each) and then washed with DCM (4X).

WO95130642 r.~ .,,s.

b) Nitrogen elaboration In the first of the seven 250 mL synthesis vessels containing dti~,.u~ ed resin from Step 4(a) (6 g, ca 3.4 mmol) is placed DCM
(150 mL) and triethylamine (15 equiv.). Phenylsulfonyl chloride (ca 5 10 equiv.) is added and the resin agitated for 4 hr. The resin is washed with ~It~-m~tin~ DCM and methanol (5X each) and then with EtOAc (2 X).
In the second of the seven 250 mL synthesis vessels containing deprotected resin from Step 4(a) (6 g, ca 3.4 mmol) is 10 placed DCM (150 mL) and triethylamine (15 equiv.). Butryl chloride (ca 10 equiv.) is added and the resin agitated for 4 hr. The resin is washed with ~Itt~rnzltin~ DCM and methanol (5X each) and then with EtOAc (2X).
In the third of the seven 250 mL synthesis vessels 15 cu~ ini-.g deprotected resin from Step 4(a) ~6 g, ca 3.4 mmol) is placed DMF (150 mL) and HOBt (ca 15 equiv.). 4-Carboxy-~.,,...f~ r,--, ",i~ (ca 10 equiv.) is added and the resin agitated for 30 min. DIC (ca 10 equiv.) is added and the resin agitated for 4 hr.
The resin is washed with alternating DCM and methanol (5X each) and then with EtOAc (2X).
In the fourth of the seven 250 mL synthesis vessels containing deprotected resin from Step 4(a) (6 g, ca 3.4 mmol) is placed DMF (150 mL) and acetic acid (3 mL). Benzaldehyde ~ca 50 equiv.) is added and the resin agitated for 30 min. Sodium ~;ya~lobOluhydride (ca 50 equiY.) is added and the resin agitated for 16 hr. The resin is washed with ~It~rn~tin~ DCM and methanol (5X each) and then with EtOAc (2X).
In the fifth of the seven 250 mL synthesis vessels containing d~yluL~ d resin from Step 4(a) (6 g, ca 3.4 mmol) is placed DMF (150 mL) and acetic acid (3 mL). Butyraldehyde (ca 50 equiv.) is added and the resin agitated for 30 min. Sodium cyanoborohydride (ca 50 equiv.) is added and the resin agitated for 16 hr. The resin is washed with altemating DCM and methanol (5X each~
and then with EtOAc (2X).
The resin from the sixth of the seven 250 mL synthesis vessels containing d~lJ-uL~L~d resin from Step 4(a) (6 g, ca 3.4 mmol) 2 1 8 ~ 6 3 4 WO 95/30642 P~ J..,5.'C', I^

is lldn~r~l,. d to a 250 mL round bottom flask. THF (150 mL) is added followed by DBU (ca 40 equiv.). 2-Chloropyrimidine (ca 20 equiv.) is added. The mixture s heated to 55C for 16 hr. The resin is transferred back to a 250 mL synthesis vessel, washed with ~ltrrn~tin~
5 DCM and methanol (SX each), and then with EtOAc (2X).
In the seventh of th~ seven 250 mL synthesis vessels ~UIILdillillg deprotected resin from Step 4(a) (6 g, ca 3.4 mmol) is placed absolute ethanol (150 mL ). Methyl iSO~ alla~ (ca 15 equiv.) is added and the resin agitated for 12 hr. The resin is washed with 0 ~It~rn~tin~ DCM and methanol (5X each) and then with EtOAc (2X).
c) Encoding of Step 4 Each of the seven ~)atches of resin from Step 4(b) are binarily encoded in a fashion an~logous to tha~ described for the encoding of Step 1.
The seven batches ~f encoded resin are combined in a 2 L
Erlenmeyer flask along with TH~ (I L) and mi~ed thoroughly by swirling and stirring gently with a glass rod. The resin is then recovered by filtration and vacuum dried. This resin is then divided into three batches of 0.7 g each a}ld seven batches of 5.7 g each. The 20 seven 5.7 g. batches are subjecte~ to Step 5. The three 0.7 g batches are subjected to alternate Step 5.
Step 5 a) Encoding Each of the seven 5.7 g. batches of resin from Step 4(c) 25 and the seven 1.4 g batches from Step 3(b) are binarily encoded in a fashion analogous to that described for the encoding of Step 1.
b) Reductive amination The seven encoded 5.7 g. batches of resin from Step 5(a) are placed in 200 mL round bottom flasks. The seven encoded 1.4 g.
30 batches from Step 3(c) are placed in 50 mL round bottom flasks. To each of the fourteen flasks is added a solution of 10% glacial acetic acid in methanol (60 mL in the 200 mL flasks, 15 mL in the 50 mL flasks).
The d~ idt~ amine from Table 2-6 (ca 40 equiv.) is added followed by sodium cyanoborohydride (ca 40 equiv.). Condensers are attached 30642 F~ . ''G' I~

and the mi%tures are heated to 75C for 48 hr. The resin is washed with alternating DCM and methanol (5X each) and then with EtOAc (2X).
Alternate Step 5 a) Thiokct~1i7~ n One of the three 0.7 g. batches of resin from Step 4(c) and one of the 0.2 g batches of resin from Step 3(c) are placed in two separate 30 mL synthesis vessels. To each is added DCM (6 mL), followed by 1,2-~ f~ -1 (I mL) and boron trifluoride etherate (I
mL). The resin is agitated at r.t. for 6 hr. The resin is washed with DCM (IX) and then treated once more with c~ (l mL) and boron t~ifllln~ etherate (I mL). The resin is agitated at r. t. for 14 hr. The resin is then filtered and washed with s~lf~rn:~fin~ DCM and methanol (5X each) and then with EtOAc (2X).
b) Reduction One of the three 0.7 g. batches of resin from Step 4(c) and one of the 0.2 g batches of resin from Step 3(c) are placed in two separate 30 mL synthesis vessels. To each is added methanol (6 mL) and (cautiously) solid sodium bolohyd.ide (200 mg). The flasks are vented and allowed to gently shake for 1 hr. The resin is filtered and resuspended in methanol (6 mL) and the reduction process repeated a total of S times at 1 hr. intervals using 200 mg portions of sodium borohydride each time. After the final cycle, the resin is washed with ~1tt~7T~tin~ DCM and methanol (5X each) and then with EtOAc (2X).
c) One of the three 0.7 g. batches of resin from Step 4(c) and one of the 0.2 g batches of resin from Step 3(c) is left unaltered.
Step 6 a) Mixing 30 The seven 5.7 g batches of encoded resin from Step S(b) are combined in a 2 L Erlenmeyer flask along with THF (1 L). The seven 1.4 g batches of encoded resin from Step S(b) are combined in a 500 mL Erlenmeyer flask. Each batch of resin is mixed thoroughly by i~
Wo 95r?~0642 ~ ' 2 1 8 9 6 3 4 r~ r ~ c~

swirling and stirring gently with a glass rod. The resin from each flask is recovered by filtration. vacuum dried, and kept separate.
b) Nitrogen elaboration.
The mixed and dried resin from the combined 5.7 g.
5 batches in Step 6(a) (total of ca 32.5 g.) is divided into ten 3.2 g.
batches and placed in 100 mL synthesis vessels. The mixed and dried resin from the combined 1.4 g. batches in Step 6(a) (total of ca 9.8 g.) is divided into ten 0.98 g. batches and p~aced in 30 mL synthesis vessels.
These vessels are paired up into ten sets of two where each set has one 10 100 mL vessel and one 30 mL vessel. Both members of each set are subjected to the same reaction conditions as outlined below.
In the first set of vessels is placed N,N'-bis Boc-(L)-lysine (ca 10 equiv.) as a solution in DMF (60 mL in the larger vessel, 15 mL
in the smaller). HOBt (c~ 15 equiv.) is added and the resin agitated for lS IS min. DIC (ca 10 equiv.) is added and the resin agitated for 4 hr.
The resin is washed with DCM (2X) and then treated with TFA (30%
solution in DCM) (1.5 hrs.). The resin is then washed with DCM (2X) and treated with 10% tricillyl~l~lille in DMF (2X, 30 min. each). The resin is washed with alternating DCM and methanol (5X each) and then 20 with EtOAc (2X).
In the second set of vessels is placed N-a-Fmoc-N-~-Pmc-(L)-arginine (ca 10 equiv.) as a solution in DMF (60 mL in the larger vessel, 15 mL in the smaller). HOBt (ca. lS equiv.) is added and the resin agitated for 15 min.. DIC (~. 10 equiv.) is added and the resin agihted for 4 hr. The resin is washed with DCM (2X) and then treated with TFA (50% solution in DCM) (l S hrs.). The resin is then washed with DCM (2X) and treated with 50% piperidine in DMF (2X, 30 min.
each). The resin is washed with alternating DCM and methanol (5X
each) and then with EtOAc (2X).
In the third set of vessels is placed DCM (60 mL in the larger vessel, IS mL in the smaller). N,N-di-n-propyl-N'-cyano-ethylthioformamidine (ca. 15 equiv.) is added, followed by triethylamine (ca 20 equiv.) and the resin agitated for 12 hrs. The resin is washed with :~ltern~tin~ DCM and MeOH (5X each) and then with EtOAc (2X).

~, WosS/30642 ' '' ` ~ 89634 r~ Jri~ ~c In the fourth set of vessels is placed absolute ethanol (60 mL in the larger vessel, 15 mL in the smaller). Methyl isocyanate (ca IS equiv.) is added and the resin agitated for 12 hr. The resin is washed with Alf~rnAting DCM and methanol (SX each) and then with S EtOAc (2X).
In the fifth set of vessels is placed absolute ethanol (60 mL
in the larger vessel, 15 mL in the smaller). Methyl isothiocyanate (ca IS equiv.) is added and the resin agitated for 12 hr. The resin is washed with Altf~rn~tin~ DCM and methanol (5X each) and then with 10 EtOAc (2X).
In the sixth set of vessels is placed absolute ethanol (60 mL
in the larger vessel, lS mL in the smaller). Phenyl isocyanate (ca. lS
equiv.) is added and the resin agitated for 12 hr. The resin is washed with AltPrn~tin~ DCM and methanol (SX each) and then with EtOAc lS (2X).
- In the seventh set of vessels is placed absolute ethanol (60 mL in the larger vessel, 15 mL in the smaller). Phenyl isothiocyanate (ca. 15 equiv.) is added and the resin agitated for 12 hr. The resin is washed with Al~PrnAtin~ DCM and methanol (SX each) and then with EtOAc (2X).
In the eighth set of vessels is placed DCM (60 mL in the larger vessel, 15 mL in the smaller) and 2,6-lutidine (ca. 20 equiv.).
Isopropyl chloroformate (ca. 15 equiv.) is added and the resin agitated for 4 hr. The resin is washed with AltPmAfin~ DCM and methanol (SX
each) and then with EtOAc (2X).
In the ninth set of vessels is placed DCM (60 mL in the larger vessel, 15 mL in the smaller) and triethylamine (15 equiv.).
Isobutryl chloride (ca. 10 equiv.) is added and the resin agitated for 4 hr. The resin is washed with alternating DCM and methanol (SX each) and then with EtOAc (2X).
~n the tenth set of vessels is placed DCM (60 mL in the larger vessel, 15 mL in the smaller) and triethylamine (15 equiv.).
MethAnPslllfonyl chloride (ca. 10 equiv.) is added and the resin agitated for 4 hr. The resin is washed with AltcrnAtin~ DCM and methanol (SX
35 each) and then with EtOAc (2X).

~ $- 2 t 8963~
W095130612 r~ O
-12~ .
Table 2-1 (CH_)L~R17 Reagents and Encodin~ Scheme ~N~oX
HO_~ Me HO~
0 C15Cl l 0 c15cc7l ~ N~ oX ~ N~ oX
HO_~ ~ HO~ ~
ClsC8 ClsC7 1~1 Cl5C8 ~~OMe OMe ~ N Ox 7 ~ X
ClsC~
Cl5C
4 ~ X
O Cl5C~ ~l OMe ~ 2 1 ~ 9 6 3 4 WO 95/30642 F~ JJ~O~

Table 2-2 Substituted Hvdroxvacetophenone Rea~ents . HOJ~'O`~D~ r~
~ OH ~ OH
OH

O ~f O
3 ~O~ 'X;
O Me HO~f ~ Me O
Table 2-3 R_R~ Step 3 Rea~nt~
O O O
1 ~) 2 1~ ~N~r ~+ ~
t t~l~t t:~ 2189634 WO 95/30642 P~ J,.. .'t 1^

Table 2-4 ~R- Step 3 Alt. Rea~ents O
5 =C
5 2 Ji OMe 6 O=CS
3 J~ N~ Me- IN~
7 ~

4 O=C) Table 2-5 R8 Rea~ents O
~ SO2CI 5 ~l~ H
f~
2 ~~ Cl 6 N~ N
Cl ~ O 7 Me-N=C=O
H2NO2S ~OH

H

T ~ . 2 ~ 8 9 6 3 4 Table 2-6 (CH,~)~14 Rea~ents 5 1 5 [~3 ~--NH2 f NH2 6 ~3 MeO ~~ N~'} ~
3 7 ~3 NH2 D/` NH2 Table 2-7 B~7 ~P~pl?ntC
HS SH 2 NaBH4 3 No Reaction BF3oEt2 WO 95/30642 P~ I I 1 ~,3. ~.~, I ---12~
Table 2-8 __ Rea~ents NHBOC /=\
5 1 ~(CH2)3--NHBOC 6 ~N=C=O
NHFmoc ~H ~N=C=S

3 EtS J~ N ~ >_ o_lL Cl 4 Me-N=C=O O
9 ~CI
Me-N=C=S 10 Me- SO2CI

2~ .

Claims (34)

WHAT IS CLAIMED IS:

1. A compound of the formula:
(T'-L)q-(S)-C(O)-L'-II' I
wherein:
(S) is a solid support;
T'-L- is an identifier residue;
-L'-II' is a ligand/linker residue; and q is 3-30.

2. A compound of Claim 1 wherein:
T'-L- is of the Formula:
III
wherein n = 3-12 when Ar is pentachlorophenyl and n = 3-6 when Ar is 2,4,6-trichlorophenyl;
q is 4-12; and -L'- is or (a) (b) wherein the left-hand bond as shown is the point of attachment to the solid support and the right hand bond is the point of attachment to the ligand, and B is O or N(CH2)1-6R17, with the proviso that in (b) when B is N(CH2)1-6R17, the ligand is attached to B through a carbonyl group.

3. A compound of Claim 1 wherein -C(O)-L'-II' is:
Ia Ib or Ic

4. A compound of the formula:
II
wherein:
R1 is OH, O(CH2)1-2OH, OCH2CO2H, CO2H, O-Z-C(O)NH(CH2)1-6R17, or OCH2-4-Phe-C(O)NH(CH2)1-6R17;
R2 is H or lower alkyl;
R3 is H, alkyl, aryl, or arylalkyl;
R4 and R5 is each independently H, lower alkyl, or substituted lower alkyl where the substituents are 1-3 alkoxy, aryl, substituted aryl, carboalkoxy, carboxamido, or diloweralkylamido; or R4 and R5 taken together are -(CH2)n-, -(CH2)2-O-(CH2)2-,-CH2-O-(CH2)3-, -(CH2)2-NR8-(CH2)2-, -CH2-NR8-(CH2)m-, -(CH2)2CH(NHR8)(CH2)2-, -(CH2)2-S(O)0-2-(CH2)2-, or -CH2CH(N-loweralkyl)(CH2)2CHCH2-;
one of R6 and R7 is H and the other is H, OH, or N(CH2)1-6R14R15; or R6 and R7 taken together are , , , , , or ;
R8 is H, COOR9, CONHR10, CSNHR11, COR12, SO2R13, lower alkyl, aryl lower alkyl, heteroaryl, or heteroaryl lower alkyl, wherein aryl is optionally substituted with 1-3 subtituents selected from lower alkyl, lower alkoxy, halo, CN, NH2, COOH, CONH2, carboalkoxy, and mono- or di-lower alkylamino and wherein heteroaryl is a mono- or bicyclic heteroaromatic ring system of 5 to 10 members including 1 to 3 heteroatoms selected from O, N, and S and 0-3 substituents selected from halo, amino, cyano, lower alkyl, carboalkoxy, CONH2, and S-lower alkyl;
R9 is lower alkyl, aryl, aryl lower alkyl, heteroaryl, aryl substituted by 1-3 substituents selected from alkyl, alkenyl, alkoxy, methylene dioxy, and halo, or a 5 to 6-membered heterocyclic ring wherein the hetero atom is O or N, wherein heteroaryl is a heteroaromatic ring of 5 to 6 members including 1 to 2 heteroatoms selected from O, N, and S and 0-2 substituents selected from lower alkyl, dialkylamino, lower alkoxy, and halo;

R10 and R11 is each independently lower alkyl, aryl, aryl lower alkyl, or aryl substituted by 1-3 substituents selected from lower alkyl, halo, alkoxy, and haloalkyl;
R12 is lower alkyl, aryl, heteroaryl, aryl lower alkyl, heteroaryl lower alkyl, a 5- or 6-membered heterocyclic ring containing 1-2 heteroatoms selected from O, S, and N, a 5- or 6-membered heterocyclic ring containing 1-2 heteroatoms selected from O, S, and N lower alkyl, or aryl substituted with 1-3 substituents selected from lower alkyl, alkoxy, halo, sulfamoyl, lower alkyl sulfamoyl, cyano, and phenyl;
R13 is lower alkyl, aryl, or aryl substituted with 1-3 substituents selected from lower alkyl, alkoxy, halo, CN, and haloalkyl;
R14 is H;alkyl substituted by 1-3 alkoxy, S-loweralkyl, sulfamoyl, halo, alkylsulphonamido, or arylsulphonamido;
alkenyl; alkynyl; aryl; substituted aryl; heteroaryl;
substituted heteroaryl; heterocycloalkyl;
-CH2NR16C(O)R16; -C(O)NR16R16; -CH2OC(O)R16; or -CH2SC(O)R16;
R15 is H, alkyl, -C(O)X, -C(S)X, or-C(NCN)NR3R3;
R16 is lower alkyl. substituted lower alkyl, aryl, or substituted aryl;
R17 is H; alkyl substituted by 1-3 alkoxy, S-loweralkyl, sulfamoyl, halo, alkylsulphonamido, or arylsulphonamido;
alkenyl; alkynyl; aryl; substituted aryl; heteroaryl;
substituted heteroaryl; heterocycloalkyl;
-CH2NR16C(O)R16; -C(O)NR16R16; -CH2OC(O)R16; or -CH2SC(O)R16;
X is alkyl, aryl, arylalkyl, O-loweralkyl, or NR3R3 Z is -(CH2)1-6-, optionally substituted with 1-3 lower alkyl;
CHR2; Phe-CH2-, where Phe is optionally mono-substituted with halogen, lower alkyl, or alkoxy; or heteroarylene-(CH2)-;
m is 2 or3;
n is 4-9;
or a pharmaceutically acceptable salt thereof.

5. A compound of Claim 3 wherein R12 is sulfamoylphenyl.

6. A compound of Claim 3 wherein R12 is p-sulfamoylphenyl.

7. A compound of Claim 4 wherein:
R1 is OH, OCH2C(O)NH(CH2)1-6R14, or OCH2-4-Phe-C(O)NH(CH2)1-6R14;
R2 is H or lower alkyl;
R4 and R5 is each lower alkyl; or R4 and R5 taken together are -(CH2)5-, -(CH2)2-O-(CH2)2-, -(CH2)2-NR8-(CH2)2-, -(CH2)2-CH(NHR8)(CH2)2-, -(CH2)2-S-(CH2)2-, or -CH2CH( HCH2-;
R6/R7 are H/OH, =O, or -S(CH2)2S-;
R8is H, COOR9,CONHR10, CSNHR11, COR12, SO2R13,.
lower alkyl, aryl lower alkyl, heteroaryl wherein the ring members include 1 to 3 N atoms and the substituents are halo or amino, heteroaryl lower alkyl wherein heteroaryl is 6-membered and the heteroatoms are N, or aryl lower alkyl substituted with 1 substituent selected from lower alkyl, alkoxy, and halo;
R9 is lower alkyl, aryl lower alkyl, aryl, tetrahydrofuranyl, tetrahydropyranyl, or aryl substituted by 1 to 2 substituents selected from lower alkyl, alkenyl, alkoxy, methylene dioxy, and halo;
R10 and R11 is each independently aryl, aryl lower alkyl, or aryl substituted by 1 substituent selected from lower alkyl, halo, alkoxy, trifluoromethyl, and pentafluoroethyl;
R12 is lower alkyl, aryl, aryl lower alkyl, heteroaryl lower alkyl wherein the heteroatoms are N, a 5- or 6-membered heterocyclic ring containing 1-2 heteroatoms selected from S and N lower alkyl, or aryl substituted with 1 substituent selected from loweralkyl, alkoxy, halo, sulfamoyl, cyano, or phenyl;

R13 is lower alkyl, aryl, or aryl substituted with 1 substituent selected from lower alkyl, alkoxy, and halo;
or a pharmaceutically acceptable salt thereof.

8. A compound of Claim 4 of the formula:
IIa wherein:
R1 is 6- or 7-OH;
R2 is H or lower alkyl;
R4 and R5 is each methyl; or R4 and R5 taken together are -(CH2)5-, -(CH2)2-O-(CH2)2-, -(CH2)2-NR8-(CH2)2-, -CH2-NR8-(CH2)3-, -CH2-NR8-(CH2)2-, or-(CH2)2-CH(NHR8)(CH2)2-;
one of R6 and R7 is H and the other is OH or R6 and R7 taken together are =O or -S(CH2)2S-;
R8 is H, COOR9, CONHR10, CSNHR11, COR12, SO2R13, benzyl, -CH2-Ph-4-F, -CH2-Ph-4-OCH3, -CH2-4-Py, n-butyl, -CH2-c-propyl, , , , or ;

R9 is i-propyl, phenyl, phenethyl, t-butyl, , , or ;

R10 phenyl, p-chlorophenyl, or p-trifluoromethylphenyl;
R11 is phenyl, benzyl, or 1-naphthyl;

R12 is , , , , , , , , , or ; and R13 is 1-or2-naphthyl, phenyl, 4-chlorophenyl, 4-methylphenyl, 4-t-bulylphenyl, n-butyl, or i-propyl;
or a pharmaceutically acceptable salt thereof.

9. A compound of Claim 4 of the formula:
IIa wherein:
R1 is 6- or 7-OH when R2 is H;
R1 is 7-OH when R2 is CH3;
R4 and R5 is each methyl; or R4 and R5 taken together are -(CH2)5-, -(CH2)2-O-(CH2)2-, -(CH2)2-NR8-(CH2)2-, -CH2-NR8-(CH2)3-, -CH2-NR8-(CH2)2-, or-(CH2)2-CH(NHR8)(CH2)2-;
one of R6 and R7 is H and the other is OH or R6 and R7 taken together are =O or -S(CH2)2S-; and R8 is

10. A compound of Claim 4 of the formula IIa wherein:
R1 is 6- or 7-OCH2C(O)NH(CH2)1-6R17, or 6- or 7-OCH2-4 Phe-C(O)NH(CH2)1-6R17 when R is H;
R1 is 7-OCH2C(O)NH(CH2)1-6R17, or 7-OCH2-4Phe-C(O)NH(CH2)1-6R17 when R is CH3;

R4 and R5 is each methyl; or R4 and R5 taken together are -(CH2)5-, -(CH2)2-O-(CH2)2-, -(CH2)2-NR8-(CH2)2-, -(CH2)2-CH(NHR8)(CH2)2-, -(CH2)2-S-(CH2)2-, or -CH2CH(NCH3)(CH2)2CHCH2-;
or R4 is methyl and R5 is CH2OCH3 or -(CH2)3N(Et)2;
one of R6 and R7 is H and the other is OH; or R6 and R7 taken together are =O or -S(CH2)2S-; or one of R6 and R7 is H and the other is NAB, where A is methyl, 2-methoxyethyl, 2-phenylethyl, 4-~nethoxybenzyl, 2-tetrahydro-furanylmethyl, 2,3,4-dimethoxyphenyl)ethyl, or 2,2-diphenylethyl and B is H, -SO2CH3, , , , , , , , or .

R8 is H, CONHCH3, SO2Ph, (CH2)3CH3, CO(CH2)2CH3, benzyl, -C(O)-(4-Phe)-SO2NH2, or ;

R13 is , , H, -SO2CH3, , , , , , or ;

(CH2)1-6R14 is methyl, n-butyl, 3-methoxy-n-propyl, CH2-c-propyl, or -(CH2)1-3-phenyl; and (CH2)1-6R17 is methyl, 2-methoxyethyl, 2-phenylethyl, 4-methoxybenzyl, methyl-2-tetrahydrofuranyl, 2(3,4-dimethoxyphenyl)ethyl, or 2,2-diphenylethyl;
or a pharmaceutically acceptable salt thereof.

11. A compound of Claim 4 of the formulae IIb, IIc, and IId:
IIb IIc IId wherein:
R1 is 6- or7-OH, 6- or 7-OCH2C(O)NH(CH)1-6R17, or 6- or 7-OCH2-4Phe-C(O)NH(CH2)1-6R17;
R is H or CH3;
R8 is -CO-Ph-p-SO2NH2; and R6 and R7 together are =O or -SCH2CH2S-.

12. A compound of Claim 11 wherein the substituents are as follows:

13. A compound of Claim 4 of the formula:
wherein the substituents are as follows:

14. A compound of Claim 4 of the formula:
wherein the substituents are as follows:

15. A pharmaceutical composition comprising a therapeutically effective amount of a compound of Claim 4 and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition comprising a therapeutically effective amount of a compound of Claim 9 and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising a therapeutically effective amount of a compound of Claim 10 and a pharmaceutically acceptable carrier.

18. A pharmaceutical composition for inhibiting carbonic anhydrase in a mammal comprising a therapeutically effective amount of a compound of Claim 11 and a pharmaceutically acceptable carrier.

19. A method of inhibiting carbonic anhydrase isozymes in a mammal which comprises administering to said mammal an effective amount of a compound of Claim 11.

20. A method of treating glaucoma in a mammal which comprises administering to a mammal in need of such treatment an effective amount of a compound of Claim 11.

21. A method of identifying a ligand having a desired characteristic which comprises synthesizing a combinatorial library of Claim 1 and testing the compounds in said library in an assay which identifies compounds having the desired characteristic.

22. A method of of Claim 21 wherein the compounds in said library are those wherein -C(O)-L'-II' is of the formula Ia.

23. A method of Claim 21 further comprising determining the structure of any ligand so identified.

24. A method of Claim 21 wherein said characteristic is carbonic anhydrase inhibition.

25. A method for identifying compounds that are inhibitors of carbonic anhydrase which comprises preparing a mixture of 20-300 pmol test compound and aqueous solutions (total volume: 25-100 µL) of 0.03-1.2 µM carbonic anhydrase and 0.04-1.6 µM
dansylamide, exposing said mixture to U.V. (preferably 274 nm) light, and determining the amount of emitted U.V. (preferably 454 nm) light.

26. A method of Claim 25 wherein the total volume is about 50 µL, the carbonic anhydrase concentration is about 0.3 µM, the dansylamide concentration is about 0.6 µM, the mixture is exposed to 274 nm U.V. light, and the emitted U.V. light is 454 nm.

27. A method for identifying compounds that are enzyme inhibitors which is a lawn assay which comprises contacting a first layer which is a colloidal matrix containing enzyme, which matrix has embedded therein a mono-layer of solid supports with attached ligands, with a second layer which is contains a substrate which can be monitored photometrically during its enzymic conversion to product, eluting said ligands by exposure to U.V. light, and detecting zones of inhibition in said first layer produced thereby.

28. A method of Claim 27 wherein said first layer is an agarose matrix containing bovine carbonic anhydrase and said second layer is a fluorescein diacetate-containing layer of agarose.

29. A method of identifying a ligand having a desired characteristic which comprises synthesizing a combinatorial library of Claim 1, detaching the ligands from the solid supports in said library, and testing said library of ligands in an assay which identifies compounds having the desired characteristic.

30. A method of Claim 29 further comprising determining the structure of any ligand so identified.

31. The use of divinylbenzene-cross-linked, polyethyleneglycol-grafted polystyrene beads as the solid supports for constructing combinatorial libraries of Claim 1.

32. A method of Claim 31 wherein the ligand is detached from said solid supports by photolysis.

33. A process for preparing a compound of the formula:
3 where R2 is H or lower alkyl;

which comprises a) reacting allyl or methyl 4-(hydroxymethyl)-3-nitrobenzoate with a compound of the formula:
in the presence of triphenylphosphine, toluene, and DEAD and stirring the mixture at room temperature to produce where R is allyl or methyl and b) when R is allyl reacting said compound with methylene chloride, tetrakistriphenylphosphine palladium(0), and pyrrolidine and stirring the mixture at 0°C, or when R is methyl reacting said compound with dilute NaOH and THF and stirring the mixture at 0°C.

34. A compound of the formula:
wherein:
? is a solid support;
R16 is lower alkyl, substituted lower alkyl, aryl, or substituted aryl;
R17 is H; alkyl substituted by 1-3 alkoxy, S-loweralkyl, sulfamoyl, halo, alkylsulphonamido, or arylsulphonamido;
alkenyl; alkynyl; aryl; substituted aryl; heteroaryl;
substituted heteroaryl; heterocycloalkyl; -CH2SC(O)R16;
-CH2NR16C(O)R16; -C(O)NR16R16; or -CH2OC(O)R16;
R is H or alkyl; and Y is aryl or heteroaryl.