CA2286867A1 - Combinatorial libraries of peptidomimetic macrocycles and processes therefor - Google Patents

Abstract

The present invention relates to a novel diverse library of peptidomimetic macrocyclic compounds, processes for the preparation thereof, and to apparatus providing a readily accessible source of individual members of the library. The apparatus can be used in assay kits and as a replaceable element in automated assay machines.

Description

-~1-COMBINATORIAL LIBRARIES OF PEPTIDOMIMETIC
MACROCYCLES AND PROCESSES THEREFOR
Field of the Invention . This Application claims the benefit of U.S. Provisional Application No. 60/043,532, filed April 11, 1997.
The present invention relates to diverse libraries of peptidomimetic organopeptide macrocyclic compounds, combinatorial methods of making such libraries, and an apparatus for storing and providing a readily accessible source of such libraries. The apparatus harboring the present combinatorial libraries is a useful component of assay systems for identifying compounds for drug development.
Background of the Invention Research and development. expenses account for a large outlay of capital in the pha~__°maceutical industry.
Synthesis of compounds is an expensive and time consuming phase of research and development. Historically, research chemists individually synthe:~ized and analyzed high purity compounds for biological scrE:ening to develop pharmaceutical leads. Although such methods were successful in bringing new drugs to the market, the limitations of individual synthesis and complete compound characterization considerable slowed the discovery of new pharmaceutically active compounds. The need for more rapid and less expensive drug discovery methodology is increasingly important in today's competitive pharmaceutical industry.

Recently, modern drug discovery has utilized combinatorial chemistry to generate large numbers (102 -106) of compounds generally referred to as "libraries".
An important objective of combinatorial chemistry is to generate a large number of novel compounds that can be screened to identify lead compounds for pharmaceutical research.
Theoretically, the total number of compounds which may be produced for a given library is limited only by the number of reagents available to form substituents on the variable positions on the library's molecular scaffold.
The combinatorial process lends itself to automation, both in the generation of compounds and in their biological screening, thereby greatly enhancing the opportunity and efficiency of drug discovery.
Combinatorial chemistry may be performed in a manner where libraries of compounds are generated as mixtures with complete identification of the individual compounds postponed until after positive screening results are obtained. However, a preferred form of combinatorial chemistry is "parallel array synthesis", where individual reaction products are simultaneously synthesized, but are retained in separate vessels. For example, the individual library compounds can be prepared, stored, and assayed in separate wells of a microtiter plate, each well containing one member of the parallel array. The use of standardized microtiter plates or equivalent apparatus, is advantageous because such an apparatus is readily accessed by programmed robotic machinery, both during library synthesis and during library sampling or assaying.
Combinatorial chemistry can be carried out in solution phase where both reactants are dissolved in solution or in solid phase where one of the reactants is covalently bound to a solid support. Typically, completion of the solution phase reactions in combinatorial chemistry schemes are ensured by selecting high yielding chemical reactions and/or by using one reagent in considerable excess. When one reagent is used in excess, completion of the reaction produces a mixture of a soluble product with at least one soluble unreacted reagent. Solid phase synthe:~is offers the advantage that the solid support-bound products are easily washed free of excess reagent. Solution ph<~se synthesis typically requires use of one or more reaction mixture work-up procedures to separate reaction product from unreacted excess reagent.
Combinatorial chemistry may be used at two distinct phases of drug development. In the discovery phase diverse libraries are created to find lead compounds. In a second optimization phase, strong lead compounds are more narrowly modified to find optimal molecular configurations.
Small peptides as a cla~~s of compounds possess rich diversity and potent bioactivity and thus remain a vital starting point for drug discovery. However, due to a litany of undesirable pharmacokinetic properties of peptides, peptidomimetics are frequently used to redeploy important peptide main chain and side chain pharmacophores onto less peptide-like scaffolds. Peptidomimetics vary greatly in their resemblance to peptides. Success has been achieved with peptides containing hydrolytically stable amide surrogates, mixed organo-peptide frameworks, and with non-peptide scaffolds alike. Benzodiazepines, penicillins and cephalosporins are well-established peptidomimetic drugs and it is easy to see that organo-peptide chimeras are essential to both rational design and combinatorial chemistry approaches to drug discovery.
Conformational restriction is important in drug - pharmacology and efficacy because i) less flexible molecules usually bind with much higher affinity to their targets because less energy i;> expended in freezing out non-active conformations and ii) conformationally restricted peptides are usually less susceptible to digestion by proteases, hence are expected to be more active drugs. Thus, preparation of cyclic compounds has been an important drug discovery tactic to take advantage of those features. [See Hruby, Life Sciences, 31, 189-199 (1982); Kates et al., Tet Lett 34, in 1549-52 (1993);
Kataoka et al., Biooolvmers 32, 1519-33 (1992); Szewczuk et al., Biochemistry 31, 9132-40 (1992); Schiller et al., J. Med Chem., 34 3125-32 (1992); Toniolo, Int. J. Pent Prot. Res., 35, 287-300 (1990); Veber et al., Nature, 280, 512-4 (1979)]
Barker et al. [J. Med. Chem. 35, 2040-8 (1992)] and McDowell and Gadek [JACS, 114, 9245-9253, (1992)] have made constrained cyclic peptides having a single thioether bond (at the point of ring closure) and bearing the sequence Arginine-Glycine-Aspartic acid, a high affinity ligand for glycoprotein IIb-IIIa, an important receptor in the blood clotting cascade. Wen and Spatola have synthesized and characterized linear pseudopeptide libraries bearing 1 to 3 dialkylamino replacements "[i~rCH2NH]~~ for defined amide bonds [Abstract #60, Division of Medicinal Chemistry, American Chemical Society 207th National Meeting in San Diego, CA, March 13-17, 1994.]
For other reports of linear and cyclic peptides bearing a single thioether replacement for an amide bond, see Paladino et al., Int. J. Pept Prot Res. 42, 284-93 (1993); Anwer et al., Int. J. Pept Prot Res., 36, 392-9 (1990); Spatola et al., Colloa. INSERM, 174, 2nd Peptide Forum, 45-54 (1989); and Darlak et al., Pept. Proc Eur Pent. Sump 20th, 634-6 (1989), June and Gayer, Eds.]
Summary of the Invention The present invention is directed to the construction of a series of novel structurally related macrocyclic WO 98!46631 PCT/US98/07135 compounds from one or more a~minothioether compounds or one or more of their corresponding sulfoxide or sulfone derivatives, collectively referred to herein as "ATAs".
ATA compounds are used in combination with other ATAs and other organic compounds that bear amine and acid functionality, to generate the present novel, diverse macrocyclic compounds having at least 2 sulfur linkages in the macrocycle ring structure, wherein the oxidation state of those linkages may vary. By linking ATAs together in sequence, one may generate an oligomer with a backbone alternating between sulfur and amide linkages. Moreover, the flexibility of the present process for preparing the macrocyclic libraries enables construction of peptidomimetics that are not constrained to the normal spacing or "register" found in normal peptides. The process also enables facile synthesis of macrocycle libraries having the same number or a different number of ring atoms of from about 12 to about 40 ring atoms.
Libraries of these macrocyclic ATA compounds are screened to identify lead compounds through their biological activity.
In their simplest form, ATAs are dipeptides where a thioether bridge replaces th~~ central amide linkage, such as, for example, those described by Spatola, A.F., in Chemistry and Biochemistry o.f Amino Acids, Peptides and Proteins, B. VJeinstein, Ed.; Marcel Dekker: New York, 1983; Vol. 7; pages 267-357. A facile synthesis of such compounds from commercially available amino acids or amino alcohols and mercapto acids provides a diverse array of ATA compounds for use in preparation of novel macrocycle libraries described in the present invention. The diverse ATA compounds are used in combination with other organic compounds that bear amine anti acid functionality, including both proteiogenic ~~mino acids and non-proteiogenic amino acids, to construct diverse libraries of peptidomimetic macrocyclic compounds of the present invention.
The present invention is also directed to a simple and direct process for the production of diverse libraries of novel macrocyclic organo-peptide compounds useful in the identification of new pharmaceutical lead compounds.
For example, as depicted in Scheme I below, the present library compounds may be easily prepared using solid phase synthesis techniques via carbodiimide-mediated coupling of one or more ATAs with proteiogenic and non-proteiogenic amino acids, followed by solid phase or solution phase cyclization by a thioetherforming intramolecular nucleophilic displacement reaction. The anchor residue and the terminal bromoacid, as well as the identity, number and coupling order of the intervening amino acid and ATA components all contribute to the unique structural identity of the macrocycle product.
;.>
a' b' c' FmocHN --> cys --~ ~yS ---- cys 1s 2s 3s 4s BrA
Cys ~ AA2 Cys ATA AAA ATA
7s 6s 5s Scheme I
Steps: a') N-Fmoc-rink amide MBHA resin, 30% piperidine in DMF, then 5 eq Fmoc-S-trityl-L-cysteine, diisopropyl carbodiimide/hydroxybenzotriazole (DIC/HOBT) in N-WO 98/46631 PCT/tJS98/07135 methylpyrrolidinone (NMP); ~;teps b') through d') as for Step a') but with the respective amino acid (AA) or ATA;
e') 30o piperidine in DMF, then 5eq bromoacetic acid and DIC in NMP;
f') 38:1:1 trifluoroacetic acid (TFA):H20:Et3SiH
for 2 hours, lyophilize, the 2 eq of diisopropylethylamine in 1:1 CH3CN:H20.
The library is created, stored, and used as an apparatus comprising a two-dimensional array of reservoirs, each reservoir containing a predetermined library reaction product differing from those in adjacent reservoirs.
Another embodiment of the present invention provides an assay kit for the identification of pharmaceutical lead compounds, said kit comprising assay materials and a well plate apparatus or equivalent apparatus providing a two-dimensional array of defined reservoirs. The well plate apparatus provides a diverse combinatorial library, wherein each well (reservoir) contains a unique macrocycle compound, or stereoisomers and/or regioisomers thereof.
The well plate apparatus is 'used to provide multiple reaction zones for making the library, to store the library and to provide a rea~~ily accessible source of library compounds.
Brief Description of the Drawincrs Fig. 1 is a top view of a well plate in accordance with this invention.
Fig. 2 is a side view o~= a well plate apparatus for use in the process of this invention.
Detailed Descrit~tion of the Invention The term "assay kit" as used in accordance with the present invention refers to an assemblage of two cooperative elements, namely (1) a well plate apparatus and (2) biological assay materials.

_g_ "Biological assay materials" are materials necessary to conduct a biological evaluation of the efficacy of any library compound in a screen relevant to a selected disease state.
A "library" is a collection of compounds created by a combinatorial chemical process, said compounds having a common scaffold with one or more variable substituents.
A "library compound" is an individual reaction product, a single compound or a mixture of isomers, in a combinatorial library.
A "Lead compound" is a library compound in a selected combinatorial library for which the assay kit has revealed significant activity relevant to a selected disease state.
A "diverse library" means a library where the substituents on the combinatorial library scaffold or core structure, are highly variable in constituent atoms, molecular weight, and structure, and the library, considered in its entirety, is not a collection of closely related homologues or analogues (compare to "directed library").
A "directed library" is a collection of compounds created by a combinatorial chemical process, for the purpose of optimization of the activity of a lead compound, wherein each library compound has a common scaffold, and the library, considered in its entirety, is a collection of closely related homologues or analogues to the lead compound (compare with "diverse library").
The term "scaffold" as used in accordance with the present invention comprises a peptidomimetic macrocycle bearing at least two sulfur linking groups. The scaffold may be further derivatized using conventional combinatorial techniques.
"Substituents" are chemical radicals which are bonded to or incorporated onto the scaffold through the combinatorial synthesis process. The different functional groups account for the diversity of the molecules _.g_ throughout the library and are selected to impart diversity of structure, function and biological activity to the scaffold in the case of diverse libraries, and optimization of a particular biological activity in the a 5 case of directed libraries.
"Reagent" means any chemical compound used in the combinatorial synthesis process to incorporate substituents on the scaffold of a library.
"Parallel array synthesis" refers to the method of conducting combinatorial chemical synthesis of libraries wherein the individual combinatorial library compounds are separately prepared and storf=d without prior and subsequent intentional mixing.
"Simultaneous synthesis"' means making of library compounds within one production cycle of a combinatorial method (not making all libra~_y compounds at the same instant in time).
The "reaction zone" refers to the individual vessel location where the combinatorial chemical library compound preparation process of the invention is carried out and where the individual library compounds are synthesized.
Suitable reaction zones include, but are not intended to be limited to the individual wells of a well plate apparatus.
"Well plate apparatus" refers to the structure capable of holding one or more library compounds in dimensionally fixed and defined positions.
"Non-interfering substit.uents" are those chemical radicals that do not significantly impede the process of the invention and yield stable aminothioether macrocyclic library compounds.
"Aryl" means one or more aromatic rings, each of 5 or 6 ring carbon atoms and includes substituted aryl having one or more non-interfering substituents. Multiple aryl rings may be fused, as in naphthyl, or unfused, as in biphenyl.

"Alkyl" means straight or branched chain or cyclic hydrocarbon having 1 to 20 carbon atoms.
"Substituted alkyl" is alkyl having one or more non-interfering substituents.
"Halo" means chloro, fluoro, iodo or bromo.
"Heterocycle" or "heterocyclic radical" means one or more rings of 5, 6 or 7 atoms with or without unsaturation or aromatic character, optionally substituted with one or more non-interfering substituents, and at least one ring atom which is not carbon. Preferred heteroatoms include sulfur, oxygen, and nitrogen. Multiple rings may be fused, as in quinoline or benzofuran, or unfused as in 4-phenylpyridine. Suitable substituents on the heterocyclic ring structure include, but are not limited to halo, C1-C1p alkyl, C2-C10 alkenyl, C2-C1p alkynyl, C1-C10 alkoxy, C~-C12 aralkyl, C~-C12 alkaryl, C1-C10 alkylthio, arylthio, aryloxy, arylamino, C3-C10 cycloalkyl, C3-C10 cycloalkenyl, di(C1-C10)-alkylamino, C2-C12 alkoxyalkyl, C1-C6 alkylsulfinyl, C1-C1p alkylsulfonyl, arylsulfonyl, aryl, hydroxy, hydroxy(C1-C10)alkyl, aryloxy(C1-C10)alkyl, C1-C10 alkoxycarbonyl, aryloxycarbonyl, C1-C10 alkanoyloxy, aryloyloxy, substituted alkoxy, fluoroalkyl, nitro, cyano, cyano(C1-C10)alkyl, C1-C1p alkanamido, aryloylamido, arylaminosulfonyl, sulfonamido, amidino, carbamido, , carboxy, heterocyclic radical, nitroalkyl, and -(CH2)m-Z-(C1-C10 alkyl), where m is 1 to 8 and Z is oxygen or sulfur.
"Organic moiety" means a substituent comprising a non-interfering substituent covalently bonded through at least one carbon atom. Suitable substituents onto a connecting carbon atom include, but are not limited to hydrogen, halo, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkoxy, C~-C12 aralkyl, C~-C12 alkaryl, C1-C10 alkylthio, arylthio, aryloxy, arylamino, C3-C10 cycloalkyl, C3-C10 cycloalkenyl, di(C1-C1p)-alkylamino, C2-C12 alkoxyalkyl, C1-C6 alkylsulfinyl, C1-C10 alkylsulfonyl, arylsulfonyl, aryl, hydroxy, hydroxy(C1-C1p)alkyl, aryloxy(C1-Cl0)alkyl, C1-C10 alkoxycarbonyl, aryloxycarbonyl, C1-C10 alkanoyloxy, aryloyloxy, substituted (C1-C10) alkoxy, fluoroalkyl, nitro, cyano, cyano(C1-Cl0)alkyl, C1-C10 alkanamido, aryloylamido, arylaminosulfonyl, sulfonamido, amidino, carbamido, protected amino, protected carboxy, protected amino, carboxy, heterocyclic radical, nitroalkyl, and -(CH2)m-Z-(C1-C10 alkyl), where m is 1 to 8 and Z is oxygen or sulfur.
The term "amino acid" ass used in accordance with the present invention includes the 20 proteiogenic amino acids encoded by the genetic code, as well as hydroxyproline, alpha-aminoisobutyric acid, sarcosine, citrulline, cysteic acid, t-butylglycine, t-buty:Lalanine, phenylglycine, cyclohexylalanine, beta-alan:ine, 4-aminobutyric acid and other compounds of the general formula:
HN ~! COOH
Rn wherein Rn is hydrogen or an organic moiety, and Q is an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S; or Rn taken together with Q and the nitrogen atom to which they are bound form a 4 to 7-membered ring.
"Proteiogenic amino acids" are those amino acids of the formula Rn wherein R1 is selected from the group consisting of hydrogen, methyl, isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, sulfhydrylmethyl, 2(methylthio)ethyl, benzyl, 4-hydroxybenzyl, WO 98/46631 PCT/US98/0'7135 3-indolylmethyl, carboxymethyl, 2-carboxyethyl, carbamidomethyl, 2-carbamidoethyl, 4-aminobutyl, 3-guanadinylpropyl and 4-imidazolylmethyl, and Rn is hydrogen, or Rn and R1 taken together with the common bonded nitrogen atom form a pyrrolidine ring.
"Activated acid" or "activated acid group" refers to a carboxylic acid that has been reacted to form a group -C(O)X wherein X is a leaving group subject to nucleophilic displacement by nucleophiles. Exemplary of the group X is chloro (an acid chloride), -OC(O)Rx (an anhydride), or -ORx (an active ester). Preparation of such activated acid derivatives and their use in preparation of other acid derivatives are well known in the art "Acid reactive groups" refer to those nucleophilic groups capable of reacting with activated acids to form a covalent bond. Exemplary of acid reactive groups are -OH, -SH or -NHRr, where Rr is hydrogen or an organic moiety, and stabilized carbon anions.
"Solid support" refers to a solvent insoluble substrate having acid reactive groups for forming cleavable covalent bonds with acid reagents, such as S-protected amino-protected mercapto amino acids for use in preparing the present library compounds.
The term "ring atoms" in defining this invention refers to those atoms or "ring members" covalently bonded serially, one to another, to form a ring structure.
A diverse library of macrocyclic compounds is provided in accordance with the present invention. The macrocycle library embodied as an apparatus of this invention serves as a readily accessible source of diverse macrocyclic compounds for use in identifying new biologically active macrocyclic compounds through pharmaceutical and agricultural candidate screening assays, for use in studies defining structure/activity relationships, and/or for use in clinical investigation.

According to the present invention, there is provided structurally related macrocyclic compounds of the formula B
. ~C O
Rz Rz S

L
~ L.
C
O
wherein L is a divalent group o:E the formula 1 lCH2 ) o-1-C6H4CH-~?
or -CHRa- wherein Ra is hydrogen or an organic moiety;
RZ is hydrogen or methy:L;
B is hydroxy, or a group of the formula -NRyRt wherein Ry is hydrogen, alky=L, aryl or heterocycle, and Rt is hydrogen, a solid support, or Rt is a group of the formula T "_- N E
Ry"
wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and F:y~~ is hydrogen or a substituent derived from an electrophilic reagent; or Ry and Ry~~ taken together with the atoms to which they are bonded form a 6- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or Ry or Ry~~ taken together with -T- and the atoms to which they are commonly bonded form a 4- to 7-membered ring; or Ry and Ry~~ each taken together with T and the nitrogen atom to which they are respectively bonded form a 5- to 7-membered ring;
Z is a divalent group of the formula a(~1)a1-(~2)a2-(ATA1)t1-(AA3)a3-(ATA2)t2-(AA4)a4-(ATA3 ) t3- (~5 ) a5- (~6 ) a6b wherein each of AA1-6 is independently a divalent group of the formula O

I I

I

Rn and each of ATA1-3 is independently a divalent sulfur-linked group of the formula (~~r, O
II
Rn' Rs and wherein a1, a2, a3, a4, a5, a6, t1, t2, and t3 are independently 0 or 1, provided that t1 + t2 + t3 - 1, 2 or 3; and provided further that -Z- is selected to provide a 12- to 40-membered macrocycle ring of Formula I;
wherein in the above formulas for AA1_6 and ATA1-3, W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;
Q and Q' are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;

m is 0, 1 or 2;
Rs is hydrogen or an or~~anic moiety; and Rn' is hydrogen, or an organic moiety, or Rn taken together with Q and the atom: to which they are bonded form a 4- to 7-membered ring,;
Rn~ and Rs taken together with -Q'- and the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or Rn~ or Rs taken together with -Q'- and the atoms to which they are commonly bonded form a 4- to 7-membered ring.
The library compounds of this invention have a molecular weight of about 200 to about 1500, more typically about 250 to about 1250. The present macrocycle library compounds as represented by Formula I include 12 to 40 ring atoms. In one emb~~diment the library compounds are constructed to have 12 to 24 ring atoms, at least two of which are sulfur. The rin~~ atoms are derived from four types of reactants during the solid phase synthesis: 1) a solid support-anchored orthogonally protected trifunctional (protected thio:L, protected amino, carboxy) compound (viz. cysteine or penicillamine); 2) aminoacids, selected from proteiogenic amino acids and non-proteiogenic amino acids; 3) ATAs (aminothioether acids and their corresponding sulfo~s:ide and sulfone derivatives); and 4) alpha-halo acids, preferably alpha-bromo acids or carboxy- or carboxymethyl-substituted benzyl bromides. Generally, m.acrocycle synthesis is initiated by deprotecting the support anchored amine functional group. The linear (or at least acyclic) macrocycle precursor is synthesized by serial peptide coupling of at least one ATA a:nd zero, one or more amino acids. The resulting terminal amine functionality is finally coupled, by peptide bond formation, to an alpha-halo acid or a carboxy/carboxymethyl substituted benzyl bromide to provide an electrophilic (bromomethyl) terminus. Upon deprotection of the support- bound thiol and treatment with a non-nucleophilc base, the acyclic precursor undergoes intramolecular thioether-forming cyclization. Accordingly, the support bound orthogonally protected cysteine or penicillamine contributes 4 ring atoms to the macrocycle structure, use of an alpha-halo acid contributes two ring atoms, and use of a carboxy/carboxymethyl substituted benzyl bromide contributes 4 to 7 ring atoms to the macrocycle structure.
The remaining ring members are determined by the nature, coupling order and number of the amino acids and ATAs coupled during synthesis of the acyclic support bound macrocycle precursor.
The libraries in accordance with this invention can be synthesized to have diversity in the number of ring atoms, or they can be prepared so that each member of the library has the same number of ring atoms with diversity in the library being introduced by the nature and coupling order of the component amino acids (AA1-6) and ATAs.
Additional elements of diversity can be introduced into the present macrocycle library by the preparation and use of solid supports having selected acid reactive groups such as, for example, those acid reactive groups derived from reaction of diamines with p-nitrophenylcarbonate Wang resin. Trifluoroacetic acid mediated cleavage of the synthesized macrocycles or acyclic macrocycle precursors from such diamine reacted resin provides a "side chain"
having amine functionality that can be reacted with a wide variety of electrophilic reagents.
In one embodiment of the invention there is provided a diverse library of macrocycle compounds of Formula I
wherein B is NH2. Such library compounds are derived from the corresponding library compounds wherein B is a solid support, particularly a Rink-Amide AM resin or Rink-Amide MBHA resin by trifluoracetic acid mediated cleavage of the covalently bound library compound or its acyclic precursor from the solid support.
Another embodiment of the present invention provides macrocycle library compounds of Formula I wherein B is a group of the formula -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle and Rt is hydrogen, a solid support, or Rt is a group of the formula 'I' --- N E
Ry"
wherein T, Ry~~ and E are as defined above. The compounds where E is hydrogen can be d~srived from corresponding library compounds wherein E :is a solid support, particularly a Wang resin, b~~r trifluoroacetic acid mediated cleavage, or they c<~n be derived by cyclization of the corresponding acyclic polymers. The library compounds of Formula I wherein E is hydrogen are useful as core structures for the synthesis of directed macrocycle libraries having diversity in the group E through reactions with various electrophilic reagents to construct variable "side chain" functionality on the macrocycle library compounds.
The library compounds of: Formula I wherein the group B or E is a covalently bonded solid support (and the corresponding acyclic precur:~or compounds) also represent embodiments of the present irwention. A general compilation of solid support~~ (resins) can be found in "Supports for Solid Phase Organic Synthesis," Martin Winter, pp 465-510, in Combinatorial Peptide and Non-peptide Libraries, edited by Gunther Jung (1996), VCH
Publishers (Weinheim, Germany). Many solid supports having acid reactive groups, such as hydroxy and primary or secondary amino groups (or precursors thereto) are commercially available. The acid reactive groups covalently bound to solid supports are typically designed to have selectively cleavable covalent bonds linking it to the solid support. Illustrative of commercially available solid supports include, but are not intended to be limited to, the following resins:
Wang resin (where the sphere represents commercial divinylbenzene-crosslinked polystyrene):
Wang resin functionalized with various amino acids (many varieties are commercially available):
~a~R
NIHFawo Wang p-nitrophenylcarbonate resin:
NO=

~o Rink Amide AM resin:
FmooHN OMo H I
N' ~
OMo O
O
Rink Amide MBHA resin:
FmeWIH OIM
O OAAo H O

_.1g-Knorr Resin:
_b ~ I
H ~ 1 ~ G 0W
The library compounds cf Formula I of this invention wherein B is the group -NRyRt, Rt is a group of the formula T' N E
Ry and wherein T and Ry~~ are as defined above and E is a solid support, are derived from the modified Wang resin bearing acid reactive groups of the formula O
HN T N CO-CH2-CsH4-O-CHZ-C6Hd--Polymer Ry Ry"
wherein Ry, T, and Ry~~ are as defined above. The modified Wang resin is prepared by reacting Wang p-nitrophenylcarbonate resin with an excess of diamine of the formula HI T IH
Ry Ry"
Exemplary of diamine compounds suitable for preparing modified Wang resins for use in preparing library compounds of this invention wherein E is a solid support or hydrogen (after cleavage) include, but are not intended to be limited to the following:
1, 2 -DIAMINOCYCLOHEX,D.NE
1,3-BIS(AMINOMETHYL)CYCLOHEXANE
D-CYSTINE
L-ARGININE
2,6-DIAMINOPIMELIC i~CID
2,6-DIAMINOPIMELIC ~~CID

WO 98/46b31 PCT/US98/07135 N-EPSILON-CBZ-L-LYSINE
N-EPSILON-ACETYL-L-LYSINE
N-(3-AMINOPROPYL)CYCLOHEXYLAMINE
ETHYLENEDIAMINE-N, N'-DIACETIC
ACID
N,N'-DIBENZYLETHYLENEDIAMINE
PIPERAZINE

2,6-DIMETHYLPIPERAZINE
1-(2-AMINOETHYL)PIPERAZINE
4,4'-BIPIPERIDINE DIHYDROCHLORIDE
4-(AMINOMETHYL)PIPERIDINE
1,4-BIS(3-AMINOPROPYL)PIPERAZINE
HOMOPIPERAZINE
ACETALDEHYDE-AMMONIA TRIMER
ACETALDEHYDE-AMMONIA TRIMER
M-XYLYLENEDIAMINE
1,10-DIAMINODECANE
1,12-DIAMINODODECANE
N-ISOPROPYLETHYLENEDIAMINE
N-ETHYLETHYLENEDIAMINE
TETRAETHYLENEPENTAMINE
TRIETHYLENETETRAMINE
2-(2-AMINOETHYLAMINO)ETHANOL
DIETHYLENETRIAMINE
N-(N-PROPYL)ETHYLENEDIAMINE
N-(2-AMINOETHYL)-3-AMINOPROPYLTRIMETHOXYSILANE
TRIS(2-AMINOETHYL)AMINE
N-(2-AMINOETHYL)-1,3-PROPANEDIAMINE
3,3'-IMINOBISPROPYLAMINE
1,4-DIAMINOBUTANE
1,5-DIAMINOPENTANE
1,6-HEXANEDIAMINE
1,7-DIAMINOHEPTANE
1,8-DIAMINOOCTANE
1,9-DIAMINONONANE
N,N'-DIETHYL-2-BUTENE-1,4-DIAMINE
P-XYLYLENEDIAMINE
D-ARGININE MONOHYDROCHLORIDE
(+/-)-3-AMINOPIPERIDINE
DIHYDROCHLORIDE
2,2'-OXYBIS(ETHYLAMINE) DIHYDROCHLORIDE
CYSTAMINE DIHYDROCHLORIDE
D-ORNITHINE HYDROCHLORIDE
D-LYSINE MONOHYDROCHLORIDE
AGMATINE SULFATE
2-METHYL-1,5-DIAMINOPENTANE
ISOPHORONEDIAMINE

-~21-2-N-BUTYLAMINOETHS'LAMINE
1,4-DIAMINOCYCLOHE~XANE
S-(2-AMINOETHYL)-I~-CYSTEINE
HYDROCHLORIDE
N,N-DIETHYLDIETHYL~ENETRIAMINE
TERT-BUTYL N-(6-AMINOHEXYL)CARBAMP,TE
HYDROCHLORIDE
1,4-BIS(AMINOMETHYL)CYCLOHEXANE
1,8-DIAMINO-3,6-DIOXAOCTANE
N-BENZYLETHYLENEDIAMINE

DIHYDROCHLORIDE
L-CYSTINE DIHYDROXAMATE
L-ARGININE HYDROXAMATE
HYDROCHLORIDE
DIAMINOBIOTIN
L-LYSINE HYDROXAMATE
HYDROCHLORIDE
L-LYSINAMIDE DIHYDROCHLORIDE

DIHYDROCHLORIDE
(2R,2R)-(-)-1,2-DIAMINOCYCLOHEXANE
(1S, 2S) - (+) -1, 2-DIAMINOCYCLOHEXANE
CIS-1,2-DIAMINOCYC:LOHEXANE
TRAMS-1,2-DIAMINOC'i~CLOHEXANE
L-CYSTINE
L-LYSINE
L(-)-ALPHA-AMINO-El?SILON-CAPROLACTAM
L-ARGININE MONOHYDROCHLORIDE
L-ARGININE MONOHYDROCHLORIDE
L-2,4-DIAMINOBUTYRIC ACID
DIHYDROCHLORIDE
L-ORNITHINE MONOHYI)ROCHLORIDE
L-ORNITHINE MONOHYI)ROCHLORIDE
DL-LYSINE MONOHYDROCHLORIDE
DL-LYSINE MONOHYDROCHLORIDE
L-LYSINE MONOHYDROC'_HLORIDE
DL-ORNITHINE MONOHYDROCHLORIDE
L-LYSINE DIHYDROCHI~ORIDE
DL-HOMOCYSTINE
TRAMS-2,5-DIMETHYLF'IPERAZINE
ETHAMBUTOL DIHYDROC.'HLORIDE
(1S,2S)-(-)-1,2-DIPHENYLETHYLENEDIp,MINE
(2R,2R)-(+)-1,2-DIPHENYLETHYLENEDIP,MINE
DL-CYSTINE

1,3-DIAMINOPENTANE
(4S,5S)-4,5-DI(AMINOMETHYL)-2,2-DIMETHYLDIOXOLANE
PIPERAZINE HEXAHYDRATE
(S)-(+)-2-METHYLPIPERAZINE
(S) - (+) -2-(AMINOMETHYL)PYRROLIDINE

(R)-(-)-2-METHYLPIPERAZINE

4-AMINOBENZYLAMINE, and the like.
In accordance with another embodiment of this invention there is provided a library of macrocycle compounds of Formula I wherein the divalent group Z is selected so that a1 + a2 + a3 + a4 + a5 + a6 = 0 to 6 in integral steps. In other words, one to six amino acids can be integrated into the macrocycle ring. In a related embodiment of the invention the library compounds are also defined so that t1 + t2 + t3 - 1, 2 or 3. Illustrative of the divalent group Z in those embodiments include, but are not intended to be limited to:
a ATA1 _b, a AA1-ATA1-ATA2-AA4 b, a ATA1-AA3-ATA2 b, a AA1-ATA1-ATA2-AAg-AA5 b, ~ ATA1-AA3-AA4-ATA3 b, a ATA1-ATA2-ATA3-AA5 b, a AA1-ATA2-ATA3-AA5-AA6 b, a AA3-ATA2-AAg b, a ATA2-AA4-ATA3-AA5 b, a AA1-ATA1-ATA2-ATA3 b, a AA4-ATA3-AA5-AA( b, ~ AA1-ATA1 b, 2 5 a ATA1-AA3 b, and a ATA1-AA3-ATA2-ATA3 b .

-:23-One preferred embodiment of the invention is a library of macrocyclic compounds of the formula B
- C O
S
NH
L
z C -O
wherein L is a divalent group of the formula 1 (CHZ) o_1-C6H9CH2?
or -CHRa- wherein Ra is hydrogen or an organic moiety;
RZ is hydrogen or methyl;
B is hydroxy or a group of the formula -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and Rt is hydrogen, a solid support, or Rt is a group of the formula T -"--. N E
Ry ~~
wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and R.~,~~ is hydrogen or a substituent derived from an e:Lectrophilic reagent; or Ry and Ry~~ taken together with the atoms to which they are bonded form a 4- to 7-memberec3 ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or Ry or Ry~~ taken together with -T- and the atoms to which they are commonly bonded form a 6- to 7-membered ring or Ry and Ry~~ each taken together with T and the nitrogen atom to which they are respectively bonded form a 5- to 7-membered ring;
Z is a divalent group of the formula a(AA1)-(ATA1)-(AA3)~
wherein AA1 and AA3 are independently a divalent group of the formula O
Q C
Rn and ATA is a divalent sulfur group of the formula O
N-Q'-CH-S W C
Rn' RS
wherein W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from the group consisting of O, N and S;
Q and Q' are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;
m is 0, 1 or 2;
RS is hydrogen or an organic moiety; and Rn' is hydrogen, or an organic moiety, or Rn taken together with Q and the atoms to which they are bonded form a 4- to 7-membered ring;

Rn~ and Rs taken together with -Q'- and the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or - 5 Rn~ or Rs taken together with -Q'- and the atoms to which they are commonly bonded form a 4- to 7-membered - ring.
Another embodiment of this invention provides a library of macrocycle compounds of Formula I wherein each amino acid used to form the macrocycle is a proteiogenic amino acid.
Still another embodiment: of this invention is directed to a library of macrocycle compounds of the Formula I above wherein Q and/or Q~ is a group of the formula - C -_ _.- CH 2 - CH

r 1 wherein R1 is hydrogen, or a non-interfering substituent; and R2 is hydrogen or an organic moiety; or R1 taken together with Rn forms a 4- to 7-membered ring, or R1 taken together with Rs forms a 4- to 7-membered ring; or R1 taken together with R2 forms a 3- to 6-membered ring.
In still another embodiment there is provided a diverse library of macrocycle compounds of Formula I
wherein W is a divalent organic group selected from the group consisting of -CHRg-, -~~R4R5CR6R~-, and a 5- or 6-membered aromatic ring containing 0 to 4 heteroatoms selected from O, N and S;

wherein Rg is hydrogen or an organic group;
R4, R5, and R6 are independently hydrogen or alkyl;
and R~ is hydrogen, hydroxy, protected hydroxy, amino or protected amino or substituted amino wherein the substituent is derived from an electrophilic group.
The present library compounds can be prepared using combinatorial synthesis protocols, or individual macrocycle library compounds can be prepared by standard chemical synthesis techniques and used as core structure for preparation of directed libraries.
Further in accordance with this invention there is provided a process for preparing a combinatorial library of compounds of the formula B
~C O
Rz z S
NH
L a/
bb ~~z O
wherein L, Z, RZ and B are as defined above. The process comprises the steps of:
(a) covalently bonding an amino-protected, thiol-protected starting material of the formula -:27-RZ RZ
COOH
PSS
I~THP
wherein RZ is defined as abo~re, and PS and P are thiol- and amino-protecting groups, respectively, to a solid support including acid reactive groups;
(b) removing the amino protecting group on the solid support bound starting mater~_al;
(c) coupling the resulting solid support bound thio-protected starting material with an amino-protected amino acid of the formula p N -"-" Q C OOH
Rn or an amino-protected compound of the formula ~~~m P IV Q' CH S W COOH
Rd Rs wherein in said formulas P is an amino-protecting group and, Rn, Rn~, Q, Q', Rs~ m and W are as defined above;
(d) removing the amino-;protecting group from the coupled solid support bound product;

(e) optionally repeating step (c) and step (d) one or more times using the same or a different amino-protected amino acid or amino-protected thioether acid (f) coupling the solid support bound product with an organic acid of the formula COON

X
wherein L is as defined above and X is a leaving group subject to nucleophilic displacement;
(g) removing the thiol-protecting group to enable macrocycle formation by nucleophilic displacement of the group X and cleaving the covalently bound product from the solid support; and (h) when the acid reactive group on the solid support is a covalently bound group of the formula O
HN T N CO-CH2-C6H~0-CHZ-C6H,~Polymer Ry Ry optionally reacting the resulting product of Formula I
wherein B is a group of the formula N T NH
2 5 Ry Ry ,.
with an amine reactive organic electrophilic agent;

provided that at least one of the steps (c) and (e) is carried out using an amino-protected aminothioether acid.
Each of the reaction steps, whether conducted on solid phase or in solution phase, is carried out using a dry, inert, polar, aprotic solvent at a temperature of about 0° to about 30°C. In the case of solution phase reactions, stoichiometricall:y equivalent amounts or near stoichiometric amounts of reactants are typically employed. The use of solid :phase synthesis for most of the process steps (the cyclization step can be conducted in solid phase or solution phase) allows the flexibility of using excess reagents to ~~ptimize yield without complicating product isolation. Using solid phase synthesis techniques, effective work-up to remove any excess reagents comprises wa:~hing the solid support bound product one or more times wit=h reaction solvent. Suitable solvents for carrying out the process steps include dimethylformamide, dimethylsulfide, tetahydrofuran, N-methylpyrrolidone, dioxane, ethyl acetate, diethyl ether and the like. The reaction :steps can be carried out using standard combinatorial chemistry protocols to produce arrays of the present library. compounds, or they can be carried out on larger scale using standard chemical synthesis, work-up, and product isolation and purification procedures.
The initial step in the process for preparation of the present library compounds. of formula I comprises covalently bonding to a solid support an amino-protected, thiol-protected starting material of the formula RZ RZ
COOH
PSS
NHP
wherein Rz is defined as above, and Ps 'and P are thiol-protecting and amino-protecting groups, respectively, wherein the solid support includes covalently bound acid reactive groups (e. g. hydroxy or primary or secondary amino). Suitable thiol-protecting groups include, but are not intended to be limited to, trityl(triphenylmethyl), 4-methoxyphenyl-diphenylmethyl, di(4-methoxyphenyl)phenylmethyl, and the like. Exemplary of monovalent amino-protecting groups include, but are not intended to be limited to:
9-fluorenylmethyloxycarbonyl (Fmoc), tert-butyloxycarbonyl (Boc), allyloxycarbonyl (Alloc), 2-trimethylsilylethoxycarbonyl (Teoc), biphenylisopropyloxycarbonyl (Bpoc), nitroveratryloxycarbonyl (Nvoc), (4-methoxyphenyl)diphenylmethyl (MMTr), (4,4'-dimethoxyphenyl)phenylmethyl (DMT) and benzyloxycarbonyl (Cbz or Z), and the like.
Reaction conditions for the preparation of protected amines utilizing such protecting groups and conditions for the selective removal of such groups to provide the corresponding amines are well-known in the art. See for example, "Protective Groups in Organic Synthesis" 2nd ed., by Theodora W. Greene and Peter G.M. Wuts (1991), John Wiley and Sons (New York).
Covalent coupling of orthogonally protected cysteine or penicillamine to the solid support is carried out by forming an activated form of the acid, such as an active ester, and reacting the activated form with the solid support, typically using an excess (2-10 fold) of the activated form to optimize yield of the support-coupled product. Excess reagent is 'washed from the reacted solid support.
The support-coupled product is then reacted to remove amino-protecting groups using the protecting group dependent cleavage conditions. Thus, for example, the fluorenylmethyloxycarbonyl group, a preferred amino-protecting group for solid phase synthesis, is removed by treatment of the solid support bound product with from about 20% to about 30o piper:idine in dimethylformamide.
The resulting solid support bound thio-protected starting material is then coupled with an amino-protected amino acid of the formula p N '-"' Q "- COOH
Rn or an amino-protected ATA of the formula ~~~m Q' CH- S W COOH
R~, RS
wherein P is an amino-protecting group and Rn, Rn~, Q, Q~
Rs, m and W are as defined above. The coupling reaction is carried out under conditions paralleling those described above for coupling the orthogonally protected cysteine or penicillamine to the solid support using a 2-fold excess of an activated form of the protected amino acid or ATA.
Any of a wide variety of commercially available or readily synthesized amino acids can be employed in this 5 step. The protected amino acids typically have a molecular weight of about 75 to about 800. Exemplary of such amino acid reactants (and protected derivatives thereof) suitable for use in this process include, but are not intended to be limited to, the following:

N-ALPHA-CBZ-L-ARGININE
N-ALPHA-BENZOYL-L-ARGININE
CHLOROACETYL-L-TYROSINE
DL-M-TYROSINE
L-DOPA

O-METHYL-L-TYROSINE
O-BENZYL-L-TYROSINE
L-TYROSINE

S-(TERT-BUTYLTHIO)-L-CYSTEINE HYDRATE
D-CYSTINE
S-TRITYL-L-CYSTEINE
S-METHYL-L-CYSTEINE
S-BENZYL-L-CYSTEINE
S-CARBOXYMETHYL-L-CYSTEINE
LANTHIONINE
L-ASPARTIC ACID
L-LEUCINE
DL-HOMOSERINE
L-HOMOPHENYLALANINE
DL-METHIONINE SULFOXIDE
L-METHIONINE SULFOXIMINE
D-METHIONINE
L-ETHIONINE
L-C-ALLYLGLYCINE
GAMMA-L-GLUTAMYL-L-GLUTAMIC ACID

L-GLUTAMIC ACID GAMMA-BENZYL ESTER
L-GLUTAMIC ACID
L-ARGININE
L-ALPHA-AMINOADIPIC ACID
2,6-DIAMINOPIMELIC ACID

_ ?~ 3 _ N-EPSILON-CBZ-L-LYSINE
N-EPSILON-ACETYL-L-LYSINE
N-CBZ-L-ALANINE
Z-L-SERINE
N-(TERT-BUTOXYCARBONYL)-L-PHENYLALANINE
5-BROMO-N-{CARBOXYMETHYL)ANTHRANILIC ACID
N-(4-HYDROXYPHENYL)GLYCINE
N-(TERT-BUTOXYCARBONYL)GLYCINE
N-CBZ-GLYCINE
HIPPURIC ACID

Z-L-ASPARTIC ACID
L-ASPARTYL-L-PHENYLALANINE METHYL ESTER
N-CBZ-L-GLUTAMIC ACID
N-ACETYL-L-GLUTAMIC ACID
DL-ALPHA-METHYLTYROSINE
N-CARBOBENZYLOXY-2-METHYLAI~ANINE
2-(METHYLAMINO)ISOBUTYRIC ACID
ALPHA-HYDROXYHIPPURIC ACID
ETHYLENEDIAMINE-DI(0-HYDROXYPHENYLACETIC ACID) D-(-)-p-HYDROXYPHENYLGLYCI1'dE
D-PENICILLAMINE DISULFIDE
D-TERT-LEUCINE
DL-DOPS
DL-VALINE
DL-ISOLEUCINE
D-SERINE
D-PHENYLALANINE

N-ACETYLGLYCINE
N-TRITYLGLYCINE
TRICINE
GUANIDOACETIC ACID
SARCOSINE
IMINODIACETIC ACID
ETHYLENEDIAMINE-N, N'-DIACE'I'IC ACID
D-ALLO-THREONINE
N-CBZ-S-BENZYL-L-CYSTEINE
N-ACETYL-DL-PENICILLAMINE
DL-PENICILLAMINE
DL-CYSTEINE
DL-HOMOCYSTEINE

L-CARNOSINE
DL-HISTIDINE

3,4-DEHYDRO-DL-PROLINE
3-AMINOPYRAZOLE-4-CARBOXYLIC' ACID

DL-PROLINE

DL-ALPHA-(2-THIENYL)GLYCINE

L-ABRINE
D-TRYPTOPHAN

L-PIPECOLINIC ACID
NIPECOTIC ACID
ISONIPECOTIC ACID
NITRO-L-ARGININE

DL-HOMOCYSTEIC ACID
P-AMINOOXANILIC ACID

ALBIZZIIN
DL-CITRULLINE
S-CARBAMYL-L-CYSTEINE
N-ALPHA-CBZ-L-ASPARAGINE
D-ASPARAGINE
N-CBZ-L-GLUTAMINE
L-GLUTAMINE
2,2-DIPHENYLGLYCINE

D-(-)-ALPHA-PHENYLGLYCINE

D-ALANINE

D-NORVALINE
D-NORLEUCINE

GLYCINE
DL-ISOSERINE

BETA-ALANINE

N-ALPHA-ACETYL-L-LYSINE
Z-L-VALINE
N-(4-NITROBENZOYL)-BETA-ALANINE
N-(4-AMINOBENZOYL)-BETA-ALANINE

WO 98!46631 PCT/US98/07135 (R)-(-)-N-(3,5-DINITROBEN~~OYL)-ALPHA-PHENYLGLYCINE
N-(3,5-DINITROBENZOYL)-DL--LEUCINE
(+/-)-3-AMINOADIPIC ACID
ALPHA-METHYL-DL-PHENYLALANINE
DL-2-AMINO-5-PHOSPHONOVALE;RIC ACID
DL-3-HYDROXYNORVALINE HYDF;ATE
(+/-)-INDOLINE-2-CARBOXYLIC ACID
BETA-(2-THIAZOLYL)-DL-ALANINE
L-GAMMA-CARBOXYGLUTAMIC ACID
L-2-(BENZYLOXYCARBONYLAMInfO)-4-SULFAMOYLBUTYRIC ACID
L-2-AMINO-4-SULFAMOYLBUTYR.IC ACID

SARCOSINE HYDROCHLORIDE
4-(METHYLAMINO)BUTYRIC ACID HYDROCHLORIDE
L-TYROSINE HYDROCHLORIDE
L-GLUTAMIC ACID HYDROCHLORIDE
D-ARGININE MONOHYDROCHLORIDE
N-EPSILON-METHYL-L-LYSINE HYDROCHLORIDE
L-HOMOARGININE HYDROCHLORIDE

(+/-)-1,2,3,4-TETRAHYDRO-3-ISOQUINOLINECARBOXYLIC
ACID HYDROCHLORIDE
DL-PIPECOLINIC ACID HYDROCHLORIDE

GLYCINE HYDROCHLORIDE
5-HYDROXY-DL-LYSINE HYDROCI~iLORIDE
DL-2,3-DIAMINOPROPIONIC ACID HYDROBROMIDE
DL-2,3-DIAMINOPROPIONIC ACID MONOHYDROCHLORIDE
DL-.2,4-DIAMINOBUTYRIC ACID DIHYDROCHLORIDE
D-ORNITHINE HYDROCHLORIDE
D-ORNITHINE HYDROCHLORIDE

D-LYSINE MONOHYDROCHLORIDE

N-PHENYLGLYCINE
DL-2-AMINO-3-PHOSPHONOPROP7_ONIC ACID
N-CBZ-L-PHENYLALANINE
N-(CARBOXYMETHYL)ANTHRANIL7:C ACID
N-(P-TOLUOYL)-GLYCINE
L-ASPARTIC ACID MONOPOTASSI:UM SALT
L-ASPARTIC ACID MONOPOTASSI:UM SALT
N-CARBOBENZYLOXY-L-LEUCINE
N-CARBOBENZYLOXY-L-ISOLEUCI:NE
3-AMINO-2,3-DIHYDROBENZOIC ACID HYDROCHLORIDE
FMOC-L-VALINE
N-ALPHA-FMOC-L-ISOLEUCINE
FMOC-L-TRYPTOPHAN
N-ALPHA-FMOC-L-PHENYLALANINE
N-ALPHA-FMOC-L-ASPARAGINE

FMOC-L-LEUCINE
FMOC-L-METHIONINE
N-ALPHA-FMOC-L-GLUTAMINE
FMOC-GLYCINE
N-(TERT-BUTOXYCARBONYL)-L-TYROSINE
N-EPSILON-BOC-L-LYSINE
N-(TERT-BUTOXYCARBONYL)-L-ALANINE
N-(TERT-BUTOXYCARBONYL)-L-SERINE
N-(P-TOLUENESULFONYL)-L-PHENYLALANINE
STATINE
BOC-L-ASPARTIC ACID
N-ALPHA-BOC-L-ASPARAGINE
N-ALPHA-BOC-L-LYSINE
N-ALPHA-CBZ-L-LYSINE
N-(TERT-BUTOXYCARBONYL)-L-ISOLEUCINE
N-(TERT-BUTOXYCARBONYL)-D-METHIONINE, DICYCLOHEXYLAMMONIUM SALT
BOC-L-ARGININE
DL-(2-FLUOROPHENYL)-GLYCINE
L-ALPHA-(2-AMINOETHOXYVINYL)GLYCINE HYDROCHLORIDE
N-P-TOSYLGLYCINE
BETA-GUANIDINOPROPIONIC ACID
N-(PHOSPHONOMETHYL)-GLYCINE
N-ALPHA-FMOC-D-TRYPTOPHAN
DL-3-(3,4-DIHYDROXYPHENYL)ALANINE

P-IODO-D-PHENYLALANINE
O-PHOSPHO-DL-TYROSINE
D-TYROSINE
DL-TYROSINE

D-ASPARTIC ACID
DL-ASPARTIC ACID
DL-LEUCINE
D-LEUCINE
L-HOMOSERINE
D-HOMOPHENYLALANINE
DL-HOMOPHENYLALANINE
DL-METHIONINE
L-METHIONINE
D-ETHIONINE
DL-ETHIONINE

D-GLUTAMIC ACID
D-ARGININE
D-ALPHA-AMINOADIPIC ACID
DL-ALPHA-AMINOADIPIC ACID
N-CBZ-DL-ALANINE
N-CARBOBENZOXY-DL-SERINE
DL-THREONINE
L-ALPHA-METHYLTYROSINE

L-TERT-LEUCINE
D-VALINE
L-VALINE
D-ISOLEUCINE
L-ISOLEUCINE
DL-SERINE

DL-PHENYLALANINE
L-PHENYLALANINE
L-CYSTINE
L-ALLO-THREONINE
D-THREONINE
L-THREONINE
D-PENICILLAMINE
L-PENICILLAMINE
L-CYSTEINE
L-HISTIDINE
D-PROLINE
L-PROLINE

L-HYDROXYPROLINE
BETA-(2-THIENYL)-L-ALANINE
DL-TRYPTOPHAN
L-TRYPTOPHAN

D-PIPECOLINIC ACID
DL-PIPECOLINIC ACID
L-CITRULLINE
L-ASPARAGINE
DL-ALPHA-PHENYLGLYCINE
L(+)-ALPHA-PHENYLGLYCINE
DL-ALANINE
L-ALANINE
(R)-(-)-2-AMINOBUTYRIC ACID
(S)-(+)-2-AMINOBUTYRIC ACID
DL-NORVALINE
L-NORVALINE
DL-NORLEUCINE
L-NORLEUCINE
DL-LYSINE
L-LYSINE
N-ACETYL-L-METHIONINE
N-(3,5-DINITROBENZOYL)-DL-ALPHA-PHENYLGLYCINE
(S)-(+)-N-(3,5-DINITROBENZOYL)-ALPHA-PHENYLGLYCINE
N-(3,5-DINITROBENZOYL)-L-LEUCINE
L-ARGININE MONOHYDROCHLORII)E
L-2,4-DIAMINOBUTYRIC ACID I)IHYDROCHLORIDE
L-ORNITHINE MONOHYDROCHLOR7_DE
DL-LYSINE MONOHYDROCHLORIDF;
L-LYSINE MONOHYDROCHLORIDE
N-(TERT-BUTOXYCARBONYL)-L-~~SPARTIC ACID 4-BENZYL ESTER

N-ALPHA-(TERT-BUTOXYCARBONYL)-L-GLUTAMINE
N-ALPHA-(TERT-BUTOXYCARBONYL)-L-HISTIDINE

BOC-L-METHIONINE
N-(TERT-BUTOXYCARBONYL)-L-TRYPTOPHAN
N-(TERT-BUTOXYCARBONYL)-L-VALINE
L-CYSTEINE HYDROCHLORIDE MONOHYDRATE
Z-L-TRYPTOPHAN
DL-TERT-LEUCINE
BOC-L-THREONINE
N-CBZ-L-THREONINE
N-CBZ-L-HISTIDINE
3,4-DEHYDRO-L-PROLINE
NALPHA-ACETYL-L-ASPARAGINE
L-LYSINE DIHYDROCHLORIDE
L-BUTHIONINE-(S, R)-SULFOXIMINE
P-AMINO-D-PHENYLALANINE
DL-BUTHIONINE (S, R)-SULFOXIMINE

(S)-(-)-INDOLINE-2-CARBOXYLIC ACID
N-(PHOSPHONOMETHYL)GLYCINE, MONOISOPROPYLAMINE SALT
MIMOSINE
D-HOMOSERINE
(+)-OCTOPINE

DL-CYSTINE
3-(CARBOXYMETHYLAMINOMETHYL)-4-HYDROXYBENZOIC ACID

(S)-(-)-1,2,3,4-TETRAHYDRO-3-ISOQUINOLINECARBOXYLIC
ACID
N-(GAMMA-L-GLUTAMYL)-1-NAPHTHYLAMIDE MONOHYDRATE
3-(3,4-DIHYDROXYPHENYL)-2-METHYL-L-ALANINE, SESQUIHYDRATE

N-METHYL-D-ASPARTIC ACID MONOHYDRATE

N-{4-NITROBENZOYL)-L-GLUTAMIC ACID HEMIHYDRATE

L-CYSTEIC ACID MONOHYDRATE

D-ASPARAGINE MONOHYDRATE
D-LYSINE HYDRATE
L-(+)-CANAVANINE SULFATE MONOHYDRATE
D-CYSTEINE HYDROCHLORIDE MONOHYDRATE
D-HISTIDINE MONOHYDROCHLORIDE MONOHYDRATE
DL-PENICILLAMINE ACETONE ADDUCT HYDROCHLORIDE
MONOHYDRATE

-;3 9 -L-GLUTAMIC ACID, MONOSODIUM SALT, MONOHYDRATE
3,5-DIIODO-L-TYROSINE DIHYDRATE
N-(9-FLUORENYLMETHOXYCARBONYL)-L-ALANINE MONOHYDRATE
3,5-DINITRO-L-TYROSINE MONOHYDRATE

DL-GLUTAMIC ACID MONOHYDRATE
L-CYSTEINESULFINIC ACID MONOHYDRATE
KAINIC ACID MONOHYDRATE

IBOTENIC ACID MONOHYDRATE
L-HISTIDINE MONOHYDROCHLORIDE MONOHYDRATE
DL-HISTIDINE MONOHYDROCHLO:RIDE MONOHYDRATE
DL-CYSTEINE HYDROCHLORIDE MONOHYDRATE

L-LYSINE MONOHYDRATE
L-ASPARAGINE MONOHYDRATE
DL-ASPARAGINE MONOHYDRATE
(S)-(-)-ALPHA-AMINOCYCLOHE:KANEPROPIONIC ACID HYDRATE
N-BENZYLGLYCINE HYDROCHLORIDE
2-AMINO-2-NORBORNANECARBOX'~'LIC ACID

(+/-)-TRANS-3-AZABICYCLO(3.1.0)HEXANE-2-CARBOXYLIC ACID

N-CARBOBENZYLOXY-L-TYROSINI~ HYDRATE
4-HYDROXY-2,2,6,6-TETRAMETHYL-4-PIPERIDINECARBOXYLIC
ACID, and the like.
It is to be understood that the amino acids utilized according to the present invention can also be selected from proteiogenic amino acids or optical isomers thereof.
ATAs for use in the process of this invention for preparation of macrocycle libraries are readily available by synthesis from available amino alcohols, more particularly protected-amino alcohol mesylates or tosylates, and mercapto acids.
In the first step in preparation of ATA compounds of the above formula wherein m = O, an aminothioether acid, a compound of the formula Q--CI -X
II
Rn Rs is reacted with about 2 to about 1.2 stoichiometric equivalents of a mercaptide salt of a mercapto compound of the formula O
HS--W C-OH
wherein in the above formulas P, Rn, Rs, X and W are defined above. The reaction is typically carried out in solution phase, and a solution of the compound of Formula II is added slowly to a solution of the mercaptide salt.
The mercaptide salt is most typically generated by reacting the corresponding mercaptan in solution with about two stoichiometric equivalent amounts of an alkali metal alkoxide base, for example sodium methoxide. Other mercaptide forming bases can be used (e. g. alkali metal dimsylates or hydrides), but without advantage. Examples of suitable mercapto acids for use in preparing the library compounds of this invention include, but are not intended to be limited to, the following:
THIOSALICYLIC ACID
N-ACETYL-DL-PENICILLAMINE
DL-PENICILLAMINE
2,3-DIMERCAPTOSUCCINIC ACID
MERCAPTOSUCCINIC ACID
N-(2-MERCAPTOPROPIONYL)GLYCINE
N-ACETYL-L-CYSTEINE
DL-CYSTEINE

DL-HOMOCYSTEINE

D-CYSTEINE HYDROCHLORIDE

GUANIDINOETHYLTHIOPROPANOIC ACID

L-THIOHISTIDINE

CYS-GLY
DL-THIORPHAN

4-MERCAPTOHYDROCINNAMIC' ACID
D-PENICILLAMINE
L-PENICILLAMINE
L-CYSTEINE
DL-CYSTEINE HYDROCHLORIDE
BOC-CYS-OH
GLUTATHIONE
D-CYSTEINE
MESO-ALPHA, ALPHA'-DIMERCAPTOADIPIC ACID
N-ACETYL-D-PENICILLAMINE
MERCAPTOTETRAZOLYLACETIC ACID
ALPHA-MERCAPTO-P-TOLUIC ACID

2-THIOACETIC ACID-5-MERCAPTO-1,3,4-THIADIAZOLE
N-ISOBUTYRYL-L-CYSTEINE
N-ISOBUTYRYL-D-CYSTEINE
CAPTOPRIL
5-MERCAPTO-TETRAZOLE-1-pROPIONIC ACID
SALOR S98,217-2 2,6-CSBA (2-Cl-6-merc~~ptobenzoic acid) 6-MERCAPTONICOTINIC ACI:~
MAYBRIDGE RJC 01025, and the like.
In the preparation of aminothioether acid compounds the mercaptide salt is reactE=d with a compound of the Formula II. The substituent :~ is a good leaving group subject to nucleophilic disp:Lacement by the mercaptide salt. Exemplary of suitable X groups include, but are not intended to be limited to, me~sylate, tosylate, halo, and the like. Compounds of Formula II are typically derived from an amino alcohol or protected-amino alcohol of the formula N 'Q CHOH
Rn Rs wherein P, Rn, Q and RS are as defined above. The amino alcohol is selected to have a molecular weight of about 60 to about 450, most typically about 60 to about 300. In one preferred embodiment the amino alcohol starting material is reacted with methane sulfonyl chloride to provide the corresponding mesylate in high yield using the mesylation protocol of Crossland and Servis [JOC 35, 1952-6 (1970)]. Suitable amino alcohols for use in preparing the present library compounds include, but are not intended to be limited to, the following:

3-AMINOMETHYL-3,5,5-TRIMETHYLCYCLOHEXANOL
(1R,2R)-(-)-PSEUDOEPHEDRINE
L-ADRENALINE
DL-HOMOSERINE
Z-L-SERINE

N-ACETYLETHANOLAMINE
2-(METHYLAMINO)ETHANOL
N-BENZYLETHANOLAMINE
2-(ETHYLAMINO)ETHANOL
DIETHANOLAMINE
2-(PROPYLAMINO)ETHANOL
D-SERINE
(1S,2S)-(+)-2-AMINO-1-PHENYL-1,3-PROPANEDIOL
D-ALLO-THREONINE
DIISOPROPANOLAMINE
2-AMINO-2-METHYL-1,3-PROPANEDIOL
TRIS(HYDROXYMETHYL)AMINOMETHANE
N-METHYL-D-GLUCAMINE

L-ISOLEUCINOL
L-PHENYLALANINOL

L-METHIONINOL

L-PROLINOL

D-(-)-ALPHA-PHENYLGLYCINOL
(-)-NOREPHEDRINE

(+/-)-2-AMINO-1-BUTANOL

DL-ISOSERINE

3-AMINO-1,2-PROPANEDIOL

1,3-DIAMINO-2-PROPANOL
2-(2-AMINOETHYLAMINO)ETHANOL
ETHANOLAMINE

(1S,2S)-(+)-2-AMINO-3-METHOXY-1-PHENYL-1-PROPANOL
DL-PROPRANOLOL HYDROCHLORIDE

TRIS(HYDROXYMETHYL)AMINOMETHANE HYDROCHLORIDE
DL-SERINE METHYL ESTER HYDROCHLORIDE
L-SERINE ETHYL ESTER HYDROCHLORIDE
L-PHENYLEPHRINE HYDROCHLORIDE
3-HYDROXYPIPERIDINE HYDF;OCHLORIDE
DL-OCTOPAMINE HYDROCHLOF;IDE
DL-NORMETANEPHRINE HYDRC)CHLORIDE
ETHANOLAMINE HYDROCHLORIDE

N-CYCLOHEXYLETHANOLAMINE
L -NOF;ADRENAL INE
L -ADRENAL INE B I TARTFLF,TE
D-SPHINGOSINE
N-(TERT-BUTOXYCARBONYL)-L-SERINE
N-ACETYL-DL-SERINE
L-THREONINE METHYL ESTER HYDROCHLORIDE
L-ARGININIC ACID
D-GLUCOSAMINIC ACID
L-TYROSINOL HYDROCHLORIDE
L-SERINE BENZYL ESTER HYDROCHLORIDE
2-AMINO-2-METHYL-1-PROPAI~OL HYDROCHLORIDE
2-AMINO-1,3-PROPANEDIOL OXALATE

2-AMINO-1,3-PROPANEDIOL
DL-SERINE HYDROXAMATE
L-SERINE BETA-NAPHTHYLAM:IDE
N-CBZ-D-GLUCOSAMINE
N-(TERT-BUTOXYCARBONYL)E'L'HANOLAMINE
DL-THREONINE HYDROXAMATE
SER-BETA-ALA
N-T-BOC-L-HOMOSERINE

HYDROCHLORIDE
L-THREONINAMIDE HYDROCHLORIDE
3-AMINO-2,2-DIMETHYL-1-PF;OPANOL
L-HOMOSERINE

N-T-BOC-D-SERINE
N-CARBOBENZOXY-DL-SERINE
L-LEUCINOL
L-SERINE METHYL ESTER HYDROCHLORIDE
DL-THREONINE
DL-SERINE
L-SERINE
(+)-PSEUDOEPHEDRINE
(-)-EPHEDRINE
L-ALLO-THREONINE
D-THREONINE
L-THREONINE
L-VALINOL
D-VALINOL
D-PHENYLALANINOL

L-HYDROXYPROLINE
D-PROLINOL
L-(+)-ALPHA-PHENYLGLYCINOL
(1S,2R)-(+)-PHENYL-PROPANOLAMINE
(S)-(+)-2-AMINO-1-PROPANOL
D-ALANINOL
(S)-(+)-2-AMINO-1-BUTANOL
(R)-(-)-2-AMINO-1-BUTANOL
(R)-(-)-1-AMINO-2-PROPANOL
(S)-(+)-1-AMINO-2-PROPANOL
(+)-EPHEDRINE HYDROCHLORIDE
D-MANNOSAMINE HYDROCHLORIDE
D(+)-NOREPHEDRINE HYDROCHLORIDE
NOREPHEDRINE HYDROCHLORIDE
BOC-L-THREONINE
N-CBZ-L-THREONINE
DL-ALLOTHREONINE
L-SERINAMIDE HYDROCHLORIDE
ALPHA-D-GLUCOSAMINE HYDROCHLORIDE
DL-METHIONINOL
(1S,2R)-(+)-2-AMINO-1,2-DIPHENYLETHANOL
(1R,2S)-(-)-2-AMINO-1,2-DIPHENYLETHANOL
S-BENZYL-L-CYSTEINOL
BOC-L-PHENYLALANINOL
L-HISTIDINOL DIHYDROCHLORIDE
D-(-)-THREO-2-AMINO-1-(4-NITROPHENYL)-1,3-PROPANEDIOL
ETHAMBUTOL DIHYDROCHLORIDE
(+/-}-ARTERENOL BITARTRATE SALT
S-ALPHA-HYDROXYMETHYL TYROSINE
(1S,2S)-(+)-THIOMICAMINE

HYDROCHLORIDE

N-(2-HYDROXYETHYL)CARBAMIC ACID BENZYL ESTER
N-(TERT-BUTOXYCARBONYL)-L-SERINE METHYL ESTER
(R)-(+)-3-HYDROXYPIPERIDINE HYDROCHLORIDE
(S)-TERT-LEUCINOL
N-TRITYL-L-SERINE METHYL ESTER

(+/-)-NOREPINEPHRINE L-:BITARTRATE HYDRATE
(1S,2R)-(-)-CIS-1-AMINO-2-INDANOL

2-[2-(AMINOPHENYLTHIO]B:ENZYL ALCOHOL, and the like.
Amino alcohol starting materials can also be prepared in high yield from available amino acids and protected amino acids of the formula N Q' COOH
III
Rn wherein P, Rn and Q are as defined above. Available amino acid compounds can be protected with an amino-protecting group and converted in good ~~ields to the corresponding protected-amino alcohols of t:he formula CHOH
Rn Rs For example, when Rs is hydrogen these compounds can be obtained from sodium borohydride reduction of the mixed carbonate formed, for example, between the starting acid and isobutyryl chloroformate. Any of the above exemplified amino acids are available for conversion to the corresponding amino alcohols for use in preparing amino thioether acids for use in this invention.
Where the thioether-forming reaction is carried out using a dialkali metal of a mercaptoacid in excess of a stoichiometric amount relative to the amino alcohol derived compound of Formula II above, the excess reagent frequently migrates with the thioether acid product on a flash silica column and cannot be removed by simple extraction. The excess acid can be separated from the product by dissolving the reaction mixture in ethyl acetate and adding a 2-fold molar excess of solid mercuric acetate to the solution. After 3-20 hours the resulting slurry is filtered through a celite pad, the filtrate containing the desired thioether product is subjected to flash chromatography on silica, and the chromatographed material is evaporated to provide the purified reaction product.
Reaction Scheme II below (bold-face numberals correspond to compound numerals in Scheme II) illustrates of the preparation of ATA compounds, for use in accordance with this invention using amino acid starting materials, such as, for example, amino acid 1 [Formula III, where P =
Rn = hydrogen and Q =
C

wherein R1 is hydrogen and R2 - CH2R]. With reference to Scheme II below, amino acid 1 is protected, reduced to the corresponding alcohol, and then converted to mesylate 2.
The ATA is formed by condensing the disodium salt of a mercapto acid with mesylate 2 to form 3a. The next step of the process involves exchanging protecting groups to yield a N-Fmoc-protected ATA 3b. Each of the reaction -~~7-steps proceed smoothly and c;~n be carried out on multigram scale.
R O R OMs S ' W OOOH
R S-- W~-COOH
~ a, b, c 'OH ->
- a 3b 3a P=Fmoc P=Alloc P=Alloc or Boc or Boc Scheme II
a) Allyl chloroformate or d=-t-butyl dicarbonate, Na2C03, 1:1 H20:dioxane, 4oC, 90-950, P=Alloc or Boc, respectively;
b) isobutyryl chloroformate, N-methylmorpholine (NMM) in ethylene glycol dimethy:l ehter (DME), -lSoC, then aq NaBH4, -lSoC, 65-90a;
c) CH3S02C1, TEA in THF, 4oC, 90-95%;
d) mercapto acid (1eq), NaOMe (2eq) in DMF, r.t., 65-950;
e) P= Alloc~Fmoc, PPh3 (0.25eq), Pd(PPh3)4 (0.03eq), HOAc (2eq) in THF, then Fmc>c-N-hydroxysuccinimide ester, Na2C03 in 1:1 H20:dio:~ane, 4oC to r.t., 500; P=
Boc-~Fmoc, 4N HCl in clioxane or EtOAc, 1.5h, then Na2C03 and Fmoc-OSu in 1:1 H20:dioxane, 90-950.
The aminothioether compounds can be converted to their corresponding sulfoxides (m=1) and sulfones (m=2) (included in the ATAs useful in accordance with this invention) by oxidation. The oxidation can be accomplished in high yields using stoichiometric amounts of any one of several oxidizing agents including m-chloroperbenzoic acid, period,~te, and oxone (a mixture of potassium hydrogen persulfate, potassium bisulfate and potassium sulfate). The oxidation reaction is carried out in any one of a wide variety of solvents at a temperature of about -10 C to about 30°C. The preparation of sulfoxides and sulfones by oxidation of thioethers are well known in the art.

Oxidation of a sulfide to its corresponding sulfoxide~
1). P. G. Hunt, J. I. Grayson, S. Warren and J. Durman _J.
Chem. Soc. Perkin Trans 1 (1986), p. 1939. [Uses m-CPBA (meta-chloro-peroxybenzoic acid) or sodium periodate to give desired products in 92-1000 yield].
2). D. M. Hedstrand, S. R. Byrn, A. T. McKenzie and P. L.
Fuchs J. Ora. Chem. 52 (1987), pp. 592-598.
3). C. Jouen, M. C. Lasne and J. C. Pommelet Tet. Lett.
37 (1996), pp. 2413-2416.
Oxidation of a sulfide to its corres~ondina sulfone~
4). B. M. Trost and R. Braslau J. Ora. Chem. 53 (1988), pp. 532-537. [Uses Oxone from DuPont which is "...a mixture of potassium hydrogen persulfate, potassium bisulfate and potassium sulfate...."].
Amino-protected ATAs useful in the present process have a molecular weight of about 150 to about 800. A
preferred amino-protecting group for the amino acids and ATAs used in this process is 9-fluorenyloxymethylcarbonyl.
According to the process of the present invention, following the first amino acid or ATA coupling reaction of process step (c) as described above, in process step (d) the amino-protecting group is removed from the solid support coupled product following the same procedures as described above for process step (b). Thereafter, process step (c) and process step (d) may be repeated one or more times using the same or a different amino-protected amino acid or amino-protected ATA. However, at least one of the process steps (c) and (e) is carried out using an amino-protected ATA.
In the next process step (f) for preparing the present library compounds, the support-bound product resulting from process step (d) (or process step (e), if optionally performed) is coupled with an organic acid of the formula L --COON

X
where L is as defined above and X is a leaving group subject to nucleophilic displacement. Exemplary of such acids include, but are not intended to be limited to:
bromoacetic acid alpha-bromophenylacetic acid alpha-bromopropionic acid iodoacetic acid chloroacetic acid 4-carboxybenzyl bromide 4-carboxymethylbenzyl bromide 3-carboxybenzyl bromide 2-carboxybenzyl bromide, and the like.
The coupling of the organic <~cid with the terminal amino group on the solid support bound product is carried out under essentially the same rE~action conditions and stoichiometry described above for the coupling of the protected amino acids or the protected ATAs in steps (c) and (e) above except that the hydroxybenzotriazole (HOBT) reagent typically used with <:arbodiimides to form the intermediate "active" HOBT e:~ter is not used in the coupling reaction mixture due to the fact that HOBT is sufficiently nucleophilic that it reacts to displace the terminal leaving group. The coupling reaction proceeds without added HOBT via the active ester formed by the reaction of the organic acid and the carbodiimide reactant.
In the next step of the process for the preparation .. of the present macrocycle library compounds, the support- .
bound product from step (f) above is cyclized and cleaved from the solid support. While those two reactions can be conducted in either order, i.e., cyclization before cleavage, or cleavage before cyclization, it is preferred that cyclization be conducted in solution phase after cleavage of the acyclic precursor from the solid support.
The optimum reaction conditions for cleavage of the product of step (f) or of the pre-cyclized product from the solid support are dependent on the nature of the acid reactive groups on the support. Typically such cleavage reactions are carried out by treating the solid support with trifluoroacetic acid in the presence of water, optionally in the presence of triethylsilane, conditions which also effect removal of the thio-protecting group from the acyclic precursor product. The cleavage product is isolated by separation of the "cleavage cocktail"
25 solution from the solid support and lyophilization of the "cleavage cocktail". Cyclization of the acyclic macrocycle precursor product can be efficiently carried out in solution in the presence of a non-nucleophilic base such as, for example, 2,6-lutidine, diisopropylethylamine (DIPEA), and the like.
When the acid reactive group on the solid support comprises a covalently bound group of the formula O
HN T N CO-CH2-C6H~0-CH2-CsHd-Polymer Ry Ry the product macrocycle library compounds of the invention are those of Formula I wherein B is a group of the formula --- N T NH
Ry Ry ~, wherein Ry, T and Ry~~ are as defined above. Such library compounds are optionally reacted with an electrophilic agent having a molecular weight of about 30 to about 600 to provide library compounds of Formula I wherein E is a substituent derived from an electrophilic reagent.
Suitable electrophilic ageni:s include, but are not intended to be limited to, organic halides, acyl halides, sulfonic acid esters, organohaloformates, organosulfonyl halides, organic isocyanate;~, organic isothiocyanates, aldehydes, ketones, and the like. Examples of such electrophilic agents include, but are not intended to be limited to:
3,5-bis(trifluoromethy7.)benzoyl chloride benzoyl chloride 2-bromobenzoyl chloride 2-fluorobenzoyl chloride pentafluorobenzoyl chloride 2,4-difiuorobenzoyl chloride 2,6-difluorobenzoyl chloride 2-chlorobenzoyl chloride 2,4-dichlorobenzoyl chloride 2,6-dichlorobenzoyl chloride o-acetylsalicyloyl chloride 2-methoxybenzoyl chloride 2,6-dimethoxybenzoyl chloride 2-(trifluoromethyl)benzoyl chloride o-toluoyl chloride 3-bromobenzoyl chloride 3-fluorobenzoyl chloride 3-chlorobenzoyl chloride 3,4-dichlorobenzoyl chloride m-anisoyl chloride 3,4-dimethoxybenzoyl chloride 3,4,5-trimethoxybenzoyl chloride 3,5-dimethoxybenzoyl chloride 3-ethoxybenzoyl chloride isophthaloyl chloride trimesoyl chloride 3-(trifluoromethyl)benzoyl chloride m-toluoyl chloride 3-(chloromethyl) benzoyl chloride 4-bromobenzoyl chloride 4-fluorobenzoyl chloride 4-chlorobenzoyl chloride p-anisoyl-chloride 4-ethoxybenzoyl chloride 4-n-butoxybenzoyl chloride 4-n-hexyloxybenzoyl chl~~ride 4-heptyloxybenzoyl chloride 4-biphenylcarbonyl chloride terephthaloyl chloride 4-(trifluoromethyl)benzoyl chloride 4-tert-butylbenzoyl chloride p-toluoyl chloride 4-ethylbenzoyl chloride 4-n-propylbenzoyl chloride 4-butylbenzoyl chloride 4-pentylbenzoyl chloride 4-hexylbenzoyl chloride 4-n-heptylbenzoyl chloride methyl oxalyl chloride ethyl oxalyl chloride heptafluorobutyryl chloride 2-acetoxyisobutyryl chloride pivaloyl chloride 3-chloropivaloyl chloride 2-bromopropionyl chloride 2,3-dibromopropionyl chloride 2,3-dichloropropionyl chloride o-acetylmandelic acid chloride itaconyl chloride methacryloyl chloride isobutyryl chloride 2-ethylhexanoyl chloride acetyl chloride bromoacetyl chloride chloroacetyl chloride phenoxyacetyl chloride 4-chlorophenoxyacetyl chloride methoxyacetyl chloride phenylacetyl chloride 3,3-dimethylacryloyl chloride cinnamoyl chloride fumaryl chloride ethyl malonyl chloride tert-butylacetyl chloride isovaleryl chloride undecanoyl chloride lauroyl chloride myristoyl chloride palmitoyl chloride heptadecanoyl chloride stearoyl chloride propionyl chloride 3-bromopropionyl chloride 3-chloropropionyl chloride hydrocinnamoyl chloride succinyl chloride 3-carbomethoxypropionyl chloride ethyl succinyl chloride butyryl chloride 4-bromobutyryl chloride 4-chlorobutyryl chloride valeryl chloride 5-chlorovaleryl chloride adipoyl chloride hexanoyl chloride 6-bromohexanoyl chloride pimeloyl chloride heptanoyl chloride suberoyl chloride octanoyl chloride 10-undecenoyl chloride 2-chloro-2,2-diphenylac~etyl chloride dichloroacetyl chloride alpha-chlorophenylacet5rl chloride 2-chloropropionyl chloride 2-iodobenzoyl chloride 4-iodobenzoyl chloride cyclopropanecarbonyl chloride traps-2-phenyl-1-cyclopropanecarbonyl chloride cyclobutanecarbonyl ch~_oride cyclopentanecarbonyl chloride 3-cyclopentylpropionyl chloride cyclohexanecarbonyl chloride 4-cyanobenzoyl chloride 2-furoyl chloride 1-naphthoyl chloride 2-naphthoyl chloride thiophene-2-carbonyl chloride 2-thiopheneacetyl chloride trimellitic anhydride chloride 2,6-pyridinedicarboxylic acid chloride 2-quinoxaloyl chloride 2-nitrobenzoyl chloride 3-nitrobenzoyl chloride 3,5-dinitrobenzoyl chloride 4-nitrobenzoyl chloride 3,4-dimethoxyphenylacetyl chloride 3-methyladipoyl chloride 3,5-dichlorobenzoyl chloride 2,5-difluorobenzoyl chloride 3,4-difluorobenzoyl chloride 9-fluorenone-4-carbonyl chloride 3,5-difluorobenzoyl chloride (s)-(-)-n-(trifluoroacetyl)prolyl chloride benzyloxyacetyl chloride acetoxy acetyl chloride 3-cyanobenzoyl chloride 2,5-dimethoxyphenylacet~~l chloride 3-methoxyphenylacetyl chloride WO 98/46631 PCTlLTS98/07135 iminodibenzyl-5-carbonyl chloride 2,4,6-trimethylbenzoyl chloride tetrafluorosuccinyl chloride perfluorooctanoyl chloride diphenylacetyl chloride alpha-methyl valeroyl chloride methyl malonyl chloride ethyl glutaryl chloride 5-bromovaleryl chloride methyl adipyl chloride 3-cyclohexenecarbonyl chloride 3-isocyanato benzoyl chloride 2,4,6-triisopropylbenzoyl chloride fluoroacetyl chloride 2-ethoxybenzoyl chloride piperonyloyl chloride 2,4-dimethoxybenzoyl chloride 2,3,5,6-tetrachloroterephthaloyl chloride 5-(dimethylsulfamoyl)-2-methoxybenzoyl chloride 2-(4-chlorobenzoyl)benzoyl chloride 2,2-bis(chloromethyl)propionyl chloride cinnamylidenemalonyl chloride 2-phenoxypropionyl chloride 2-phenylbutyryl chloride 2-ethylbutyryl chloride p-tolylacetyl chloride gamma-methylvaleroyl chloride 3,3-dichloropivaloyl chloride 1-methyl-1-cyclohexanecarboxylic acid chloride 2-(2,4,5-trichlorophenoxy)acetyl chloride 4-chloro-3-nitrobenzoyl chloride 4-methyl-3-nitrobenzoyl chloride 2,3-dichlorobenzoyl chloride morpholine-4-carbonyl chloride p-chlorophenylacetyl chloride bicyclo[2.2.1]heptane-2-carbonyl chloride d(-)-alpha-formyloxy-alpha-phenylacetyl chloride d(-)-alpha-phenylglycine chloride hydrochloride trifluoroacetyl chloride pentafluoropropionyl chloride hexafluoroglutaryl chloride 2-chlorocinnamoyl chloride o-methoxycinnamyl chloride 5-nitro-2-furoyl chloride 2-chlorobutyryl chloride 4-phenylazobenzoyl chloride 4-n-amyloxybenzoyl chloride 4-decylbenzoyl chloride 4-octylbenzoyl chloride dl-2-methylbutyryl chloride linolenoyl chloride linolelaidoyl chloride llh-eicosafluoroundecanoyl chloride 9h-hexadecafluorononanoyl chloride 2,3-difluorobenzoyl chloride 2-(benzoyloxymethyl)benzoyl chloride 2,2-dimethylvaleroyl chloride 3,5,5-trimethylhexanoyl chloride phenothiazine-10-carbonyl chloride 3,4-dimethyl benzoyl chloride (+)-p-(2-methylbutyl)benzoyl chloride 2,4-dichlorophenoxyacetic chloride pentadecanoyl chloride nonadecanoyl chloride neoheptanoyl chloride 9-anthracenecarbonyl chloride 2-ethoxy-1-naphthoyl chloride pyrrolidine carbonyl chloride m-(chlorosulfonyl)benzoyl chloride 2-n-propyl-n-valeroyl chloride 2-chloro-4-nitrobenzoyl chloride 2-phenoxybutyryl chloride 2-chloronicotinyl chloride 6-chloronicotinyl chloride 4-(trifluoromethoxy)benzoyl chloride 2-(trifluoromethoxy)benzoyl chloride 2,6-dichloropyridine-4-carbonyl chloride 3-chlorobenzo(blthiophene-2-carbonyl chloride 4-chloromethylbenzoyl chloride neodecanoyl chloride (phenylthio)acetyl chloride 4-carbethoxyhexafluorobutyryl chloride octafluoroadipoyl chloride 2-diazo-3,3,3-trifluoro;r~ropionylchloride 2-bromobutyryl chloride arachidoyl chloride cis-vaccenoyl chloride _ 11-eicosenoyl chloride behenoyl chloride petroselinoyl chloride palmitoleoyl chloride tridecanoyl chloride 2-chloro-5-nitrobenzoyl chloride 3-methylthiopropionyl chloride methyl 4-chlorocarbonylbenzoate anthraquinone-2-carbony:L chloride carbazole-n-carbonyl ch:Loride 2-nitrophenoxyacetyl ch:Loride 2-bromo-2-methylpropionyl chloride 2-fluoro-3-(trifluoromet:hyl)benzoyl chloride 2-fluoro-4-(trifluoromet:hyl)benzoyl chloride 2-fluoro-5-(trifluoromet:hyl)benzoyl chloride 3-fluoro-5-(trifluoromethyl)benzoyl chloride 4-fluoro-2-(trifluoromethyl)benzoyl chloride 4-fluoro-3-(trifluoromethyl)benzoyl chloride 2-fluoro-6-(trifluoromethyl)benzoyl chloride 2,3,6-trifluorobenzoyl chloride 2,4,5-trifluorobenzoyl chloride 2,4-di(trifluoromethyl)benzoyl chloride 2,6-di(trifiuoromethyl)benzoyl chloride 3-(trifluoromethoxy)benzoyl chloride m-(fluorosulfonyl)benzoyl chloride trans-1,2-cyclobutanedicarboxylic acid chloride 3-cyclohexylpropionyl chloride 4-ethyl-2,3-dioxo-1-piperazinecarbonylchloride isoxazole-5-carbonyl chloride bromodifluoroacetyl chloride erucoyl chloride 2,4,6-trifluorobenzoyl chloride dichlorochrysanthemic acid chloride isononanoyl chloride 1-adamantanecarbonyl chloride 2,5-bis(trifluoromethyl)benzoyl chloride 2,3,4-trifluorobenzoyl chloride 2,3,4,5-tetrafluorobenzoyl chloride 2,4,6-trichlorobenzoyl chloride 2,4-dichloro-5-fluorobenzoyl chloride 4-methoxyphenylacetyl chloride trans-3-(trifluoromethyl)cinnamoyl chloride 3-(dichloromethyl) benzoyl chloride 4-isocyanato benzoyl chloride heneicosanoyl chloride 2-chloroisobutyryl chloride trans-4-nitrocinnamoyl chloride 3,4,5-trifluorobenzoyl chloride 5-fluoro-2-(trifluoromethyl)benzoyl chloride 2,3,5-trifluorobenzoyl chloride 2-chloro-4-fluorobenzoyl chloride (-)-alpha-chlorophenylacetyl chloride 2-(para-tolylsulfonyl)acetyl chloride 4-methyl-4-nitrohexanoyl chloride 1-chloro-4-fluorosulfonyl-2-naphthoyl chloride 2,3-dibromo-3-phenylpropionyl chloride 2-menthoxyacetyl chloride 2-phenyl-2-(phenylsulfonyl)acetyl chloride 4,4,4-trifluorocrotonyl chloride 4,4,4-trifluorobutyryl chloride 3,4-dichloro-2,5-thiophenedicarbonyl chloride pentachlorobenzoyl chloride 4,4,7,7-tetranitrosebacoyl chloride alpha,alphal-dimethylsuccinyl chloride alpha-bromoisovaleryl chloride benzoyl chloride _. 5 ~ -oleoyl chloride methyl suberyl chloride gamma-linolenoyl chloride (-)-camphanic acid chloride S 4,4'-stilbenedicarbony7_ chloride chlorinated benzoyl ch~.oride (1r)-(+)-camphanic chloride 2-(4-nitrophenoxy)tetradecanoyl chloride 7-[(chlorocarbonyl)methoxy]-4-methylcoumarin n,n-bis(2-chloroethyl)carbamoyl chloride (s)-(-)-2-acetoxypropionyl chloride linoleoyl chloride 3-chlorotetrafluoroprop~ionyl chloride 3,4-dichloropentafluorobutyryl chloride 7h-dodecafluoroheptanoyl chloride 5h-octafluoropentanoyl chloride perfluorononanoyl chloride 3h-tetrafluoropropionyl chloride 2-bromo-2,3,3,3-tetrafluoropropanoyl chloride arachidonoyl chloride pentachloropropionyl chloride 4-decenoyl chloride tridecafluoroheptanoyl chloride undecafluorocyclohexanecarbonyl chloride 4-n-nonylbenzoyl chloride 3-(trichlorogermyl)propionylchloride 3,4,5-triiodobenzoyl chloride 2-(phenylthio)propionyl chloride 2,2,2-triphenylacetyl chloride d(-)-alpha-azido-phenyl acetyl chloride 4-azido-benzoyl chloride difluoroacetyl chloride 5-chloropyrazine-2-carbonyl chloride n-(1-naphthalenesulfony.L)-1-phenylalanyl chloride n-(4-nitrophenylsulfony:L)-1-phenylalanyl chloride n-(p-toluenesulfonyl)-1-phenylalanyl chloride dimethylmalonyl chloride methyl sebacoyl chloride 2,5-:dichloropyridine-3~-carbonyl chloride 3-(2,5 xylyloxy) propionyl chloride, and the like.
Organic Halides:
benzyl bromide alpha-bromo-o-xylene alpha-bromo-m-xylene 4-(tert-butyl)benzyl bromide alpha-bromo-p-xylene tert-butyl bromoacetate methyl bromoacetate benzyl bromoacetate ethyl bromoacetate 2-bromoacetophenone 2-bromo-2'-methoxyacetophenone 2-bromo-2',4'-dimethoxyacetophenone 2-bromo-2',5'-dimethoxyacetophenone 3-methoxyphenacyl bromide 2-bromo-4'-methoxyacetophenone 2-bromo-4'-phenylacetophenone 2-bromo-4'-methylacetophenone ethyl bromopyruvate 1-bromopinacolone 1-bromo-2-butanone 1-bromo-2,2-dimethoxypropane 1-bromo-2,2-dimethylpropane bromoacetaldehyde dimethyl acetal bromoacetaldehyde diethyl acetal 1-bromo-2-methylpropane 1-bromo-2-ethylbutane 2-ethylhexyl bromide I-bromodecane 1-bromoundecane 2-bromoacetamide iodoacetamide 4-(bromomethyl)phenylacetic acid phenacyl ester isopropyl bromoacetate 5-bromo-2-methyl-2-pentene 3,4-difluorobenzyl bromide 2,5-difluorobenzyl bromide 3,5-bis(trifluoromethyl)benzyl bromide 2-bromo-2'-nitroacetophenone 3,5-difluorobenzyl bromide 2,4-bis(trifluoromethyl)benzyl bromide 8-bromo-1-octanol 4-(bromomethyl)phenylacetic acid methyl (r)-(+)-3-bromo-2-methylpropionate 4-iodobutyl acetate 7-acetoxy-4-bromomethylcoumarin 4-bromomethyl-6,7-dimethoxycoumarin 2,4-difluorobenzyl bromide methyl 2-(bromomethyl)acrylate 3-bromopropionaldehyde dimethyl acetal.
(r)-(-)-3-bromo-2-methyl-1-propanol, and the like.
Sulfonic Acid Esters:
ethyl trifluoromethanesulfonate 2,2,2-trifluoroethyl p-toluenesulfonate 2-chloroethyl-p-toluenesulfonate 1,3-propane sultone _.5g_ 5'-tosyladenosine 1,4-butane sultone cyanomethyl benzenesulf:onate hexadecyl methanesulfonate ethyl methanesulfonate 2-chloroethyl methanesulfonate ethyl p-toluenesulfonat.e trans-2-hydroxycyclohexyl p-toluenesulfonate (2r)-(-)-glycidyl tosyl.ate (s)-(+)-2-methylbutyl methanesulfonate (s)-(+)-2-methylbutyl ~~-toluenesulfonate (s)-(+)-1-phenyl-1,2-ethanediol 2-tosylate (2r)-(-)-glycidyl 3-nitrobenzenesulfonate propargyl benzenesulfon.ate 2,2-dimethyl-1,3-dioxolan-4-ylmethyl p-toluenesulfonate (r)-(-)-2,2-dimethyl-1,3-dioxolan-4-ylmethyl p-toluenesulfonate (s)-(+)-2,2-dimethyl-1,3-dioxolan-4-ylmethyl p-toluenesulfonate 1,2:5,6-di-o-isopropylidene-3-o-(methylsulfonyl)-alpha-d-glucofuranose ethyl 1-2-((methylsulfonyl)oxy)propionate (2s)-(+)-glycidyl tosylate (2s)-(+)-glycidyl 3-nitrobenzenesulfonate 3-o-acetyl-6-o-benzoyl-5-o-(methylsulfonyl)-1,2-0-isopropylidene-alpha-d-glucofu (r)-(-)-1-benzyloxy-3-(p-tosyloxy)-2-propanol (s)-(+)-1-benzyloxy-3-(p-tosyloxy)-2-propanol ethyl 1-2-((trifluoromethylsulfonyl)oxy)propionate 2-(2-chloroethoxy)ethyl methanesulfonate 1-cyanoethyl p-toluenesulfonate, and the like.
Organohaloformates:

9-fluorenylmethyl chlor~~formate phenyl chloroformate 4-chlorophenyl chlorofo:rmate methyl chloroformate benzyl chloroformate vinyl chloroformate isobutyl chloroformate 2-ethylhexyl chloroform<~te ethyl chloroformate 2-bromoethyl chloroformate 2-chloroethyl chloroformate 1-chloroethyl chloroformate allyl chloroformate n-propyl chloroformate butyl chloroformate n-hexyl chloroformate octyl chloroformate 2,2,2-trichloro-1,1-dimethylethyl chloroformate 2,2,2-trichloroethyl chloroformate cholesteryl chloroformate 4-nitrophenyl chloroformate 4-nitrobenzyl chloroformate (-)-menthyl chloroformate 4-t-butylcyclohexyl chloroformate cetyl chloroformate (+)-1-(9-fluorenyl)ethyl chloroformate isopropyl chloroformate 3-chlorocyclohexyl chloroformate decyl chloroformate oleyl chloroformate octadecyl chloroformate butenediol bischloroformate 2-chlorobenzyl chloroformate 4-chlorobutyl chloroformate (+)-menthyl chloroformate 4,5-dimethoxy-2-nitrobenzyl chloroformate cyclopentyl chloroformate t-butylcyclohexyl chloroformate menthylchloroformate p-tolyl chloroformate 4-bromophenyl chloroformate 4-fluorophenyl chloroformate 4-methoxyphenyl chloroformate 2-nitrophenyl chloroformate 4-methoxycarbonylphenyl chloroformate 1-chloro-2-methylpropyl chloroformate (+/-)-1,2,2,2-tetrachloroethyl chloroformate 2,2-dichloroethyl chloroformate myristyl chloroformate cyclohexyl chloroformate chloromethyl chloroformate, and the like.
Organosulfonylhalides:
1-naphthalenesulfonyl chloride dansyl chloride 2-naphthalenesulfonyl chloride 2-acetamido-4-methyl-5-thiazolesulfonyl chloride 2-thiophenesulfonyl chloride B-quinolinesulfonyl chloride benzenesulfonyl chloride pentafluorobenzenesulfonyl chloride 2,5-dichlorobenzenesulfonyl chloride 2-nitroberizenesulfonyl chloride 2,4-dinitiobenzenesulfonyl chloride 3,5-dichloro-2-hydroxybenzenesulfonyl chloride 2,4 6-triisopropylbenzE~nesulfonyl chloride 2-mesitylenesulfonyl chloride 3-nitrobenzenesulfonyl chloride p-bromobenzenesulfonyl chloride 4-fluorobenzenesulfonyl. chloride 4-chlorobenzenesulfonyl. chloride 4-chloro-3-nitrobenzene~sulfonyl chloride 20 pipsyl chloride 4-nitrobenzenesulfonyl chloride 4-methoxybenzenesulfonyl chloride 4-tert-butylbenzenesulfonyl chloride p-toluenesulfonyl chloride trifluoromethanesulfonyl chloride trichloromethanesulfonyl chloride isopropylsulfonyl chloride methanesulfonyl chloride alpha-toluenesulfonyl chloride trans-beta-styrenesulfonyl chloride 2,2,2-trifluoroethanesulfonyl chloride 1-hexadecanesulfonyl chloride ethanesulfonyl chloride 2-chloroethanesulfonyl chloride 1-propanesulfonyl chloride 3-chloropropanesulfonyl chloride 1-butanesulfonyl chloride methyl 2-(chlorosulfony;L)benzoate 2-vitro-4-(trifluoromethyl)benzenesulfonyl chloride 3-(trifluoromethyl)benzenesulfonyl chloride 1-octanesulfonyl chloride 4-(trifluoromethoxy)benzenesulphonyl chloride (1r)-(-)-10-camphorsulfonyl chloride d-(+)-10-camphorsulfonyl. chloride (+/-)-10-camphorsulfonyl. chloride 2-vitro-alpha-toluenesul.fonyl chloride, and the like.
Isocyanate Reagents:
trans-2-phenylcyclopropyl isocyanate phenyl isocyanate 2-bromophenyl isocyanate 2-fluorophenyl isocyanate 2,4-difluorophenyl isocyanate 2,6-difluorophenyl isocyanate 2-chlorophenyl isocyanate 2,3-dichlorophenyl isocy.~nate 2,4-dichlorophenyl isocy,~nate 2,5-dichlorophenyl isocy~nate 2,6-dichlorophenyl isocy~~nate 2-methoxyphenyl isocyanate 2,4-dimethoxyphenyl isocyanate 2,5-dimethoxyphenyl isocyanate 2-ethoxyphenyl isocyanate 2-(trifluoromethyl)phenyl isocyanate o-tolyl isocyanate 2,6-dimethylphenyl isocyanate 2-ethylphenyl isocyanate 3-bromophenyl isocyanate 3-fluorophenyl isocyanate 3-chlorophenyl isocyanate 3,4-dichlorophenyl isocyanate 3-methoxyphenyl isocyanate 3-(trifluoromethyl)phenyl isocyanate m-tolyl isocyanate 4-bromophenyl isocyanate 4-fluorophenyl isocyanate 4-chlorophenyl isocyanate 4-methoxyphenyl isocyanate ethyl 4-isocyanatobenzoate 4-(trifluoromethyl)phenyl isocyanate p-tolyl isocyanate n-(chlorocarbonyl) isocyanate benzoyl isocyanate tert-butyl isocyanate (s)-(-)-alpha-methylbenzyl isocyanate isopropyl isocyanate methyl isocyanate ethyl isocyanatoacetate octadecyl isocyanate ethyl isocyanate 2-chloroethyl isocyanate allyl isocyanate n-propyl isocyanate butyl isocyanate cyclohexyl isocyanate 1-naphthyl isocyanate (r)-(-)-1-(1-naphthyl)ethyl isocyanate 4-fluoro-3-nitrophenyl isocyanate 2-nitrophenyl isocyanate 3-nitrophenyl isocyanate 4-nitrophenyl isocyanate 2,6-diisopropylphenyl isocyanate benzyl isocyanate 3-chloropropyl isocyanate ethoxycarbonyl isocyanate 3,5-bis(trifluoromethyl)phenyl isocyanate 2,4,6-tribromophenyl isocyanate 2,5-difluorophenyl isocyanate 2,4,5-trichlorophenyl isocyanate 2,4,6-trichlorophenyl isocyanate 2-methoxycarbonylphenyl. isocyanate 2-ethoxycarbonylphenyl isocyanate 2-isopropylphenyl isocyanate 2,3-dimethylphenyl isocyanate 4-methoxy-2-methylphenyl isocyanate 2,4-dimethylphenyl isocyanate 2,5-dimethylphenyl isocyanate 2-ethyl-6-methylphenyl isocyanate 3-cyanophenyl isocyanate 5-chloro-2,4-dimethoxyphenyl isocyanate 3-chloro-4-methylphenyl isocyanate 3,5-dichlorophenyl isocyanate 5-chloro-2-methoxyphenyl isocyanate 3,4,5-trimethoxyphenyl isocyanate 3,5-dimethoxyphenyl isocyanate 3-(methylthio)phenyl isocyanate 3-ethoxycarbonylphenyl isocyanate 3-acetylphenyl isocyanate 3,4-dimethylphenyl isocyanate 3,5-dimethylphenyl isocyanate 2-methoxy-5-methylphenyl isocyanate 3-ethylphenyl isocyanate 4-chloro-2-methoxyphenyl isocyanate 4-chloro-2-trifluoromethylphenyl isocyanate 4-chloro-3-trifluoromethylphenyl isocyanate 4-iodophenyl isocyanate 4-phenoxyphenyl isocyanate 4-ethoxyphenyl isocyanate 4-(methylthio)phenyl isocyanate 4-acetylphenyl isocyanate 4-isopropylphenyl isocy~~nate 4-ethylphenyl isocyanat~~

4-n-butylphenyl isocyan;~te 3-(dichloromethylsilyl)propyl isocyanate octyl isocyanate 4-methyl-3-nitrophenyl :isocyanate 4-chloro-2-nitrophenyl :isocyanate 2-methyl-4-nitrophenyl :isocyanate 4-methyl-2-nitrophenyl :isocyanate 2-fluoro-5-nitrophenyl :isocyanate 2-methyl-5-nitrophenyl isocyanate 3-bromopropyl isocyanatE~

2,4,6-trimethylphenyl i;;ocyanate 2-isopropyl-6-methylphenyl isocyanate 2,6-diethylphenyl isocy<~nate 5-chloro-2-methylphenyl isocyanate 4-chloro-2-methylphenyl isocyanate 4-(trifluoromethoxy)phenyl isocyanate 4-trifluoromethylthiophenylisocyanate 2,4-dibromophenyl isocy~~nate 2,6-dibromo-4-ethylphen~~l isocyanate 2,3,4,5-tetrachlorophenyl isocyanate 2-chloro-5-trifluoromethylphenyl isocyanate 2-chloro-6-methylphenyl isocyanate 2-n-carbobutoxyphenyl isocyanate 2,4,5-trimethylphenyl isocyanate 2-methyl-6-(t-butyl)phenyl isocyanate 2-ethyl-6-isopropylphenyl isocyanate 3-chloro-2-methoxyphenyl isocyanate 3-chloro-2-methylphenyl isocyanate 3-chloro-4-fluorophenyl isocyanate 4-cyanophenyl isocyanate 4-bromo-2-methylphenyl isocyanate 4-bromo-2,6-dimethylphenyl isocyanate 2,6-dibromo-4-fluorophenyl isocyanate 4-n-butoxyphenyl isocyanate 4-butoxycarbonylphenyl isocyanate phenethyl isocyanate 2-methyl-3-nitrophenyl isocyanate hexyl isocyanate hexadecyl isocyanate methylene bis(o-chlorophenyl isocyanate) 4-chloro-3-nitrophenyl isocyanate 2-chloro-4-nitrophenyl isocyanate 4,5-dimethyl-2-nitrophenyl isocyanate 2-chloro-5-nitrophenyl isocyanate 2-methoxy-4-nitrophenyl isocyanate 3-fluoro-4-methylphenyl isocyanate 5-fluoro-2-methylphenyl isocyanate 3,5-dicarbomethoxyphenyl isocyanate 2,4-dichlorobenzyl isocyanate 2-(methylthio)phenyl isocyanate n-(methoxycarbonyl)isocyanate n-(phenoxycarbonyl)isocyanate 2-biphenylyl isocyanate 3-iodophenyl isocyanate 4-phenylphenyl isocyanate tetrahydro-2-pyranyl isocyanate 4-(tert-butyl)phenylisocyanate 1-(4-bromophenyl)ethyl isocyanate isocyanatoacetic acid n-butyl ester dodecyl iqocyanate 6,7-methylenedioxy-4-isocyanate-methylcoumarin (r)-(+}-alpha-methylbenzyl isocyanate (+/-)-I-(1-naphthyl)ethyl isocyanate (s)-(+)-1-(1-naphthyl)ethyl isocyanate 3,4-difluoropher,yl isocyanate 2-methoxy-5-nitrophenyl isocyanate undecyl isocyanate ethyl 2-isocyanato-4-methyl valerate ethyl 6-isocyanatohexanoate ethyl 2-isocyanato-4-methylthiobutyrate ethyl 2-isocyanatoprop_~onate ethyl 3-isocyanatopropionate ethyl 2-isocyanato-3-methylbutyrate tert-butyl 3-isothiocyanatopropionate ethyl 2-isocyanato-3-phenylpropionate 1,3-bis(isocyanatom ethyl)cyclohexane 2-(trifluoromethoxy)phenyl isocyanate 4-(chloromethyl) phenyl. isocyanate 1-adamantyl isocyanate 1,3-bis(2-isocyanato-2-~propyl)benzene n-amyl isocyanate n-heptyl isocyanate 2-chloroethyl isocyanat.e, [ethyl-1,2-14c]
1,1,3,3-tetramethylbutyl isocyanate 3,5-dinitrophenyl isocyanate, and the like.
Aldehydes:
Ethyl 2-formyl-1-cyclopropanecarboxylate Cyclohexanecarboxaldehyde 1,2,3,6-Tetrahydrobenzaldehyde Diphenylacetaldehyde 2-Phenylpropionaldehyde 2,3-Dimethylvaleraldehyde Isobutyraldehyde 2,6-Dimethyl-5-hepten-1-al 2-Methylbutyraldehyde 2-Ethylbutyraldehyde 2-Methylpentanal 2-Ethylhexanal 2-Methylundecanal Phenylacetaldehyde Isovaleraldehyde 7-Methoxy-3,7-dimethyloctanal Undecanal Dodecanal Tridecanal Tetradecyl aldehyde Propionaldehyde 3-Phenylpropionaldehyde 3-(Methylthio)propionaldehyde Butyraldehyde Cis-4-decen-1-al N-valeraldehyde Hexanal Heptaldehyde Octanal Nonanal Decanal Undecylenic aldehyde Cis-11-hexadecenal Cis-13-octadecenal Cis-9-hexadecenal 2,5-Dimethoxy-3-tetrahydrofurancarboxaldehyde 3,5,5-Trimethylhexanal Succinic semialdehyde (+/-)-3-Phenylbutyraldehyde 2,6,6-Trimethyl-1-cyclohexene-1-acetaldehyde Cyclopropanecarboxaldehyde 3-Cyclohexylpropionaldehyde Hydroxycitronellal Cis-4-heptenal Cis-6-nonen-1-al Tetrahydrocitral Cis-7-decen-1-al Cis-8-undecen-1-al 3,5,6-Trimethyl-3-cyclohexene-1-carboxaldehyde Lyral(r) Bis(2-chlorophenyl)acetaldehyde 2-Thioglyceraldehyde 3-(4-Isopropylphenyl)isobutyraldehyde 2-Ethyl-3-methylbutanal 2-Ethylcaprylaldehyde 3-Methylvaleraldehyde 3-Phenyl-3-(p-tosyl)propionaldehyde 3-Hexenal 3-(Methylthio)butanal Veltonal Citronellal 2-(Trifluoromethyl)propionaldehyde 3,3-Dimethylbutyraldehyde Campholene aldehyde 2-Formylpropionic acid methyl ester 5-Hydroxypentanal p-Methylphenylacetaldehyde Omega-ketoheptanoic acid 4-Chlorophenylcyanoacetaldehyde Hexadecanal Methyl 7-oxoheptanoate Diethyl formyl succinate 4-Pregnene-20-beta-carboxaldehyde-3-one Cis-7-tetradecenal Cyclopentylmethanal 3,4-Dimethyl-3-cyclohexenylmethanal 2,4,6-Trimethyl-3-cyclohexen-1-carboxaldehyde Adipic semialdehyde methyl ester Cis-14-methyl-8-hexadecenal Cis-3-hexen-1-al Trans-4-decen-1-al 2,2-Dichlorooctadecanal ._67_ 2,2-Dichlorotetradecanal 2,2-Dichlorooctanal 2,2-Dichlorohexanal (r)-(+)-Citronella) 8-Methyl-7-nonenal 2-(p-Tolyl)propionaldehyde Aldehyde C-11 MOA (2-mE~thyldecanal) Alpha-methylhydrocinnarnaldehyde (s)-(-)-Citronella) 4-Hydroxybutanal 4-Oxobutyric acid meth~rl ester 3,3,4,4,5,5,5-Heptafluoropentanal 3-Methylbutanal-1-13c 6-Methyl-3-cyclohexene--1-carboxaldehyde 4-(4-Methyl-2-pentenyl)-3-cyclohexene-1-carboxaldehyde 3-Pentyn-1-al 3-Pyridylacetaldehyde rf-oxide 2,3-Dihydro-5-methoxy-~~-phenyl-2-indolecarboxaldehyde 2,4-biphenyl-3-oxobutyraldehyde 3,3,3-Triphenylpropionaldehyde 2-Bromo-n-(3-formyl-1-methylpropyl)benzamide 3-(Phenylthio)butyraldehyde Diethyl 2-(diethoxymethyl)-3-formylsuccinate 2-Chloro-3-(4-nitrophenyl)-propionaldehyde 2-Acetoxypropionaldehycle 2-Methyl-4-phenylpentan.al (lr,2s,3r,4s)-(+)-2-Ben.zyloxy-3-formyl-oxybornane 5-(4'-Chlorophenoxy)-1-pentanal Boc-ala-CHO
Boc-leu-CHO
Boc-phe-CHO
Boc-tyr(OBzl)-CHO
Boc-tyr(OMe)-CHO
Boc-val-CHO
4-Pentenal 1-Formyl-6-(dimethylamino)fulvene 1,4-Dioxaspiro(4.5)decane-7-acetaldehyde Alpha-citronella) Diethyl 2-Acetamido-2-(2-formylethyl)malonate 3,4,4,5,5,5-Hexafluoro-3-(trifluoromethyl)pentanal 3,4,4,4-Tetrafluoro-3-(heptafluoropropoxy)butanal 3,4,4,4-Tetrafluoro-3-(trifluoromethoxy)butanal 3,4,4,4-Tetrafluoro-3-(trifluoromethyl)butanal 3,3,4,4,5,5,6,6,7,7,8,8,8-Tridecafluorooctanal 3,3,3-Trifluoropropanal Beta,beta-dimethylhydrocinnamaldehyde 5-Norbornene-2-carboxaldehyde Chrysanthal 9-Decenal Decyl aldehyde, [1-14c]
4,4,4-Trifluorobutyraldehyde 3-Methyl-3-butenal 3-(5-Methyl-2-furyl)butanal 3-Phenyl-4-pentenal Tert-butyl (s)-4-formyl-2,2-dimethyl-3-oxazolidinecarboxylate Trans-2-dodecenal 9,10-Dihydro-9,10-ethanoanthracene-11-carboxaldehyde Methyl hexyl acetaldehyde 2,3-Dihydro-2-oxo-1H-imidazol-4-carboxaldehyde N-Acetylmuramic acid, and the like.
Other suitable aldehydes useful in preparation of the present libraries are further illustrated by the following formulas, wherein L is -CHO:
L\ ~ i v L
O / 1 L i L
/ 1 ~ o / ~ ' ~ o / \ i L L L
i la _ N.~ o ~ CH3 L L L
O
a / / C~ /
O
OH
OH
L L
~C/ O \
i / ~ C H
/ o H3C~O~ O~ CH3 OH
L L L
OH °' a i I \
.-- /
OH
s H,c ~
L L
L
OH
O O
1 _ I .
N
H3C~ ~ CHI
L L
HD
O OH

_7p_ L
HN, ~ I ~ I

O
L L
( ~ I I \
~c~ ~ ~ a~

Q"I C H 3 N O O~
L L
\
' /
ci I - Br F
L L
\
f i ~ c~ \

o CI CH3 L
L
\ ~ ~c o = . / ~ cH3 'o off B

L O L
i i / O ~ /
Br L L

F /
I
O
L L L
Br F nl L L L
O~N ~ ~ W /
O
O O
O\

L
F
o' _cH, F
~0 L L L
I ~ ~ ~ F
/ / /
I~C~ N~ CI~
F
L L L
i i L
CI
OH
L
L
L
n O CHs ,3 S L
HaC L

L
f"~C O CJ"~ I w I
O CHs L L L
li ' \ i H3C o L o t L
~C i ' I I _ I
HsC ~ / ~ I ~ o ~' ~ o H3C ' L
O L L
I . ' i ~ I I
'3C~° H3C ~ /
V

L
' L
L
I ~ o II

w ~C~/~/~ O
HOC O
I
L L

H3~~ J ~ ~

L L L
a I . . I C~

roc L L
L

w ~ ~u ~ ~ /J
V
i Br L
H3C~
O
/O

Ketones:

1,1,1-TRIFLUORO-2,4-PENTANEDIONE
FLUOROACETONE
4'-CHLOROACETOACETANILIDE

CHLOROACETONE
1,3-DICHLOROACETONE

CYCLOTRIDECANONE
CYCLOPENTADECANONE
DICYCLOPROPYL KETONE
CYCLOPROPYL METHYL KETONE
(-)-THUJONE
CYCLOBUTANONE

4-CYCLOPENTENE-1,3-DIONE
1,3-CYCLOPENTANEDIONE
2-METHYL-1,3-CYCLOPENTANEDIONE
CYCLOPENTANONE

2,4-DIMETHYLCYCLOPENTANONE

_75-BETA-IONONE
BETA-IONONE
ALPHA-IONONE

3-METHYL-2-CYCLOHEXEN-:1-ONE

3,5-DIMETHYL-2-CYCLOHE:KEN-1-ONE
ISOPHORONE
1,3-CYCLOHEXANEDIONE
2-ACETYL-1,3-CYCLOHEXAI~EDIONE
2-METHYL-1,3-CYCLOHEXAI~TEDIONE
DIMEDONE
1,4-CYCLOHEXANEDIONE
DIMETHYL 1,4-CYCLOHEXANEDIONE-2,5-DICARBOXYLATE
DIETHYL 1,4-CYCLOHEXANEDIONE-2,5-DICARBOXYLATE
CYCLOHEXANONE

2-(3-METHOXYPHENYL)CYCI~OHEXANONE
ETHYL 2-CYCLOHEXANONEC~~RBOXYLATE
ETHYL 4-METHYL-2-CYCLOFiEXANONE-1-CARBOXYLATE

L-MENTHONE

(+)-DIHYDROCARVONE
2,6-DIMETHYLCYCLOHEXANC)NE

3,3,5,5-TETRAMETHYLCYCI~OHEXANONE

3,4,8,8A-TETRAHYDRO-8A-METHYL-1,6(2H,7H)-NAPHTHALENEDIONE

BETA-TETRALONE

CYCLOOCTANONE
CYCLONONANONE
ALPHA-ACETYL-PHENYLACETONITRILE

4-(4-HYDROXYPHENYL)-2-BUTANONE
1,3-ACETONEDICARBOXYLIC ACID

LEVIJLINIC ACID
4,6-DIOXOHEPTANOIC ACID

4-ANDROSTENE-3,17-DIONE
CYCLODECANONE
CYCLOUNDECANONE
CYCLODODECANONE
CIS-BICYCLO(3.3.0)OCTANE-3,7-DIONE
CIS-1,5-DIMETHYLBICYCLO(3.3.0)OCTANE-3,7-DIONE

CYCLOHEPTANONE

HYDROXYACETONE

TROPINONE

1-(BETA-PHENYLETHYL)-4-PIPERIDONE

1,4-CYCLOHEXANEDIONE MONO-2,2-DIMETHYLTRIMETHYLENE
KETAL

METHYLGLYOXAL
DIMETHYL ACETYLSUCCINATE
DIMETHYL 1,3-ACETONEDICARBOXYLATE

4-(4-ACETOXYPHENYL)-2-BUTANONE

METHYLGLYOXAL DIMETHYL ACETAL
1,1-DIPHENYLACETONE
1,1-DIPHENYLACETONE

3-METHYL-2,4-PENTANEDIONE
3-ETHYL-2,4-PENTANEDIONE
ACETONE

(DIMETHYLAMINO)ACETONE
PHENOXYACETONE
METHOXYACETONE
DIMETHYL (2-OXOPROPYL)F'HOSPHONATE

3,4-DIMETHOXYPHENYLACE'TONE

METHYL VINYL KETONE
4-METHOXY-3-BUTEN-2-ONE.
4-METHOXY-3-BUTEN-2-ONE.
BENZYLIDENEACETONE
BENZYLIDENEACETONE
ACETOACETANILIDE
O-ACETOACETANISIDIDE
O-ACETOACETOTOLUIDIDE
P-ACETOACETANISIDIDE
METHYL ACETOACETATE
BENZYL ACETOACETATE
BENZOYLACETONE
ACETYLACETONE
DIACETONE ACRYLAMIDE
ACETYLACETALDEHYDE DIMETHYL ACETAL
BENZYLACETONE
4-(4-METHOXYPHENYL)-2-BUTANONE
ACETONYLACETONE

N-TERT-BUTYLACETOACETAMIDE
TERT-BUTYL ACETOACETATE
4,4-DIMETHYL-2-PENTANONE
MESITYL OXIDE
PHORONE

GERANYLACETONE
GERANYLACETONE
DIISOPROPYL KETONE

METHYL ISOBUTYL KETONE
6-METHYL-2,4-HEPTANEDIOIVE
2,6-DIMETHYL-4-HEPTANONE

DTETHYLAMINOACETONE

ETHYL 2,4-DIOXOVALERATE

DIETHYL ACETYLSUCCINATE

_78-ETHYL ISOBUTYRYLACETATE
ETHYL ACETOACETATE
DIETHYL 1,3-ACETONEDICARBOXYLATE
ETHYL LEVULINATE

ETHYL VINYL KETONE
ETHYL PROPIONYLACETATE
2,4-HEXANEDIONE

ETHYL BUTYRYLACETATE

BUTYL LEVULINATE

DIMETHYL (2-OXOHEPTYL)PHOSPHONATE

1,1-DICHLOROACETONE

_7g_ 6,7-DIMETHOXY-2-TETRALC)NE
4,4-DIMETHYL-2-CYCLOHE~;EN-1-ONE

N-(ACETOACETYL)GLYCINE
3,5-DIACETYLTETRAHYDROF'YRAN-2,4,6-TRIONE
ALLYL ACETOACETATE
4-(TRIMETHYLSILYLOXY)-3-PENTEN-2-ONE
ETHYL DIACETOACETATE
(R)-(-)-4,4A,5,6,7,8-HE~XAHYDRO-4A-METHYL-2(3H)NAPHTHALENONE
4,4-DIMETHOXY-2,5-CYCLOHEXADIEN-1-ONE

TETRONIC ACID

DI-TERT-BUTYL 1,3-ACETONEDICARBOXYLATE
1,4-CYCLOHEXANEDIONE MONOETHYLENEKETAL
DIMETHYL (3-PHENOXYACETONYL)PHOSPHONATE

5-METHYL-1,3-CYCLOHEXANEDIONE

2-CHLORO-5,5-DIMETHYL-1,3-CYCLOHEXANEDIONE

4-(TERT-BUTYLDIMETHYLSILYLOXY)-3-PENTEN-2-ONE

ETHYL 2-(TRIMETHYLSILYL:METHYL)ACETOACETATE

METHYL 4-ALLYL-3,5-DIOXO-1-CYCLOHEXANECARBOXYLATE
METHYL 2-OXO-1-CYCLOHEP'rANECARBOXYLATE
METHYL ETHYL KETONE

4-(4-HYDROXY-3-METHOXYPHENYL)-3-BUTEN-2-ONE
(S)-(+)-2,3,7,7A-TETRAH'YDRO-7A-METHYL-1H-INDENE-1,5(6H)-DIONE
2-HYDROXYMETHYL-6-METHO:?~Y-1,4-BENZOQUINONE

2,2,6,6-TETRAMETHYL-4-P:LPERIDONE HYDROCHLORIDE

HYDROCHLORIDE
ETHYL 1-BENZYL-3-OXOPIP1~RIDINE-4-CARBOXYLATE
HYDROCHLORIDE

HYDROCHLORIDE

DELTA-AMINOLEWLINIC ACID, METHYL ESTER
HYDROCHLORIDE

1,4-DIAMINO-2-BUTANONE DIHYDROCHLORIDE
DIACETONAMINE HYDROGEN OXALATE
ACETOACETIC ACID LITHIUM SALT
S-TERT-BUTYL ACETOTHIOACETATE

N-HYDROXYSUCCINIMIDYL ACETOACETATE

1,3-DIBROMOACETONE
(4-CHLOROPHENYLTHIO)PROPAN-2-ONE

2-{1-CYCLOHEXENYL}CYCLOHEXANONE
2-(BETA-CYANOETHYL)CYCLOHEXANONE

TROPONE
METHYLSULFONYLACETONE
BIS(4-METHOXYBENZYLIDENE)ACETONE
BIS(4-METHOXYBENZYLIDENE)ACETONE
3,4-DIMETHOXYBENZYLIDENEACETONE

2,6-DIMETHYL-3,5-HEPTANEDIONE

3,5-HEPTANEDIONE
2,4-OCTANEDIONE
2,4-NONANEDIONE

(METHYLTHIO)ACETONE
(+/-)-2-ETHOXYCYCLOHEXANONE

2,6-DIMETHOXY-P-BENZOQUINONE
4-(P-HYDROXYPHENYL)-3-BUTEN-2-ONE
3,4-METHYLENEDIOXYBENZYLIDENE ACETONE
3,4-METHYLENEDIOXYBENZYLACETONE
PHENYLACETONE
2',5'-DICHLOROACETOACETANILIDE

2,4,4-TRIMETHYLCYCLOPENTANONE
CYCLOHEXYLACETONE
3,3,5-TRIMETHYLCYCLOHEXANONE

__gl-ACETOACETAMIDE

3 -N-BUTYL-2 , 4-PENTANED7:ONE

SALOR 517,446-7 (BENZYLTHIO)ACETONE
6-PHENYLHEXA-3,5-DIEN-2-ONE

N,N-DIETHYLACETOACETAMIDE
ETHYL 2-(PHENYLAZO)ACETOACETATE
DIETHYL ACETOMALONATE

(ETHYLTHIO)ACETONE

N-AMYL ISOPROPYL KETONE

NOOTKATONE
CYCLOBUTYL METHYL KETONE

N,N-DIMETHYLACETOACETAM=CDE
FURFURALACETONE
ACETOACET-M-XYLIDIDE
ETHYL 2 -ETHYLACETOACETA'l'E
DICYCLOHEXYL KETONE
CYCLOHEXYL METHYL KETONE
1,2:5,6-DI-O-ISOPROPYLII)ENE-ALPHA-D-RIBO-3-HEXOFURANOSULOSE

BENZYL ISOPROPYL KETONE
1,1,3-TRICHLOROACETONE
METHYL BUTYRYLACETATE
CHLORODIFLUOROACETYLACETONE
1-(2-FURYL)-1,3-BUTANEDIONE
4,6-NONANEDIONE

N,N-DIMETHYL-2-CHLOROACETOACETAMIDE

2,6,6-TRIMETHYL-2-CYCLOHEXENE-1,4-DIONE

CYCLOPENTYLACETONE

(+/-)-2-ALLYLCYCLOHEXANONE
6-METHYL-3,5-HEPTADIEN-2-ONE
FLUORAL-P
FLUORAL-P
ISOPROPYL ACETOACETATE
N-METHYLACETOACETAMIDE
2-ETHYL-1,3-CYCLOPENTANEDIONE
3,5-DIMETHYL-4-HEPTANONE
EXO-2-CHLORO-5-OXO-BICYCLO[2.2.1]HEPTANE-SYN-ETHYL-6-(2-OXOCYCLOPENTYL)-HEXANOATE
DIETHYL (2-OXOPROPYL)PHOSPHONATE

1,1,1-TRIFLUORO-5-METHYL-2,4-HEXANEDIONE
O-(CHLOROPHENYL)ACETONE
3-AMINO-5,5-DIMETHYL-2-CYCLOHEXEN-1-ONE
PHENYL SULFONYL ACETONE
1,3-DIPHENYL-2,4-PENTANEDIONE

1-PHENYL-2,4-PENTANEDIONE

S-UNDECANONE

1,1,1-TRIFLUORO-2,4-HEXANEDIONE

3-CHLORO-5,5-DIMETHYL-2-CYCLOHEXEN-1-ONE

5-PHENYLCYCLOHEXANE-1,3-DIONE
TERT.-BUTYL-4-CHLOROACETOACETATE

3-METHYLENE-2,6-HEPTANEDIONE
(4-METHYLPHENYLTHIO)ACETONE

._g3_ 1-(THIEN-2-YL)BUT-1-EN-3-ONE

ACETOACETOXYETHYL METHACRYLATE

METHYL TRAMS-4-OXO-2-PI~NTENOATE
METHYL TRAMS-4-OXO-2-PI~NTENOATE
ISOBUTYL ACETOACETATE

BENZYL LEVULINATE
(R)-(+)-3-METHYLCYCLOHEXANONE
(R)-(+)-3-METHYLCYCLOPENTANONE

(S)-{+)-10-METHYL-1(9)--OCTAL-2-ONE
(1S)-{-)-VERBENONE
DEHYDROACETIC ACID
PINONIC ACID
PINONIC ACID
1,3-DIAMINOACETONE DIHS.'DROCHLORIDE MONOHYDRATE
(5S)-5,6-ISOPROPYLIDENE:DIOXY-6-METHYL-HEPTAN-2-ONE
N-OCTYL 4-CHLOROACETOAC.'ETATE

BICYCLO[3.2.1]OCTAN-2-ONE

BICYCLO(3.3.1)NONAN-9-ONE
BICYCLO(3.3.1)NONANE-3,7-DIONE
(1R)-(+)-NOPINONE
NORCAMPHOR
NORCAMPHOR
NERYLACETONE
4,4-DIMETHYL-2-CYCLOPEN'TEN-1-ONE
4,4-DIPHENYL-2-CYCLOHEXEN-1-ONE
THIOTETRONIC ACID
4,4-DIMETHYL-1,3-CYCLOHEXANEDIONE
2-CHLORO-1,4-BENZOQUINONE

5,5-DIMETHYL-2-PHENACYL-1,3-CYCLOHEXANEDIONE

3-(2-HYDROXYETHYLAMINO)-5,5-DIMETHYL-2-CYCLOHEXEN-ABSCISIC ACID
ABSCISIC ACID

CYCLOHEXIMIDE
PSEUDOPELLETIERINE HYDROCHLORIDE

DEHYDROCARNITINE HYDROCHLORIDE
3-(PHENYLAMINO)-CYCLOHEX-2-ENE-1-ONE

EXO-2-BROMO-5-OXO-BICYCLO[2.2.1]HEPTANE-SYN-7-CARBOXYLIC ACID
ANTI-3-OXOTRICYCLO(2.2.1.02,6)HEPTANE-7-CARBOXYLIC
ACID
(+/-)-ISOPHORONE OXIDE
PHTHALIMIDOACETONE
1-PHENYL-1,4-PENTANEDIONE
(+/-)-2-PHENYLCYCLOHEPTANONE
ACETYLMALONONITRILE

5,7-DIMETHYL-3,5,9-DECATRIEN-2-ONE

2,4-DIHYDRO-5-METHYL-2-PHENYL-4-PROPIONYL-3H-D-(-)-TAGATOSE
3-(DIMETHYLAMINO)-5,5-DIMETHYL-2-CYCLOHEXEN-1-ONE
(3AS,7AS)-(+)-HEXAHYDRO-3A-HYDROXY-7A-METHYL-1,5-INDANDIONE
(+/-)-EXO-6-HYDROXYTROPINONE
(1R-(1ALPHA,2BETA,3ALPHA))-(+)-3-METHYL-2-(NITROMETHYL)-5-OXOCYCLOPENTANEACETIC
MENTHONE

1,5-DIAMINO-3-OXAPENTANE

(1R,3S)-2,2-DIMETHYL-3-(2-OXOPROPYL)-CYCLOPROPANEACETONITRILE
(1S,3S)-3-ACETYL-2,2-DIMETHYLCYCLOBUTANE
ACETONITRILE
5,5-DIMETHYLHEXANE-2,4-DIONE

HYDROCHLORIDE

HYDROCHLORIDE
DL-3-(1-ACETOXY-1-METHYLETHYL)-6-OXOHEPTANENITRILE
DL-3-(1-METHYL-1-ETHENYL)-6-OXOHEPTANENITRILE

METHYL (1R,3S)-2,2-DIMETHYL-3-(2-OXOPROPYL}-CYCLOPROPANEACETATE
N-BENZYLPSEUDOPELLETIERINE
D-(-)-FRUCTOSE
(S)-(+)-ERYTHRULOSE HYDRATE
2,2,6,6-TETRAMETHYL-4-PIPERIDONE MONOHYDRATE

-g5-5-ISOPROPYL-1,3-CYCLOHE;XANEDIONE HYDRATE
D-(+)-SORBOSE
L- ( - ) -SORBOSE
METHYL JASMONATE
N-BENZYLTROPINONE

(+/-)-BICYCLO(3.3.1)NON.'ANE-2,6-DIONE
ANTI-5-CARBOXYTRICYCLO[2.2.1.0(2,6)]HEPTAN-3-ONE
EXO-2-CHLORO-SYN-7-HYDR.OXYMETHYL-5-OXO-BICYCLO[2.2.1]HEPTANE
L-RIBULOSE HYDRATE
4-ACETYL-2,4-DIHYDRO-5-METHYL-2-PHENYL-3H-TRICYCLO[5.2.1.02,6]DECAN-8-ONE

CARBOXYLATE
5,5-DIMETHYL-3-(METHYLAMINO)-2-CYCLOHEXEN-1-ONE
2-ACETYL-1,3-CYCLOPENTAI~EDIONE
2,4-DIMETHOXYPHENYLACETONE
2, 6-DIPHENYLCYCLOHEXANO:I~E
5-HYDROXY-2-ADAMANTANON:E

2,2'-METHYLENEBIS(1,3-C'YCLOHEXANEDIONE) (+/-)-7-OXABICYCLO(4.1.0)HEPTAN-2-ONE

2-(METHYLTHIO)CYCLOHEXA1VONE
(S)-(+)-3,4,8,8A-TETRAH'~DRO-8A-METHYL-1,6(2H,7H)-NAPHTHALENEDIONE, and the like.
Where aldehydes and ketones are utilized as the electrophilic reagent, the rE~sulting imine compounds can be reduced using sodium borohydride or other borohydride reducing agents to provide the corresponding secondary amine (Formula I; Ry~~ _ hydrogen and E is a substituent derived from an electrophilic: agent). Still further diversity can be introduced into the present library by reacting the secondary amine library compounds with an electrophilic reagent as described above to provide compounds of Formula I wherein Ry~~ and E are each independently a substituent derived from an electrophilic reagent.
Solid phase synthesis methods have been modified in accordance with the present invention to provide a general route to preparing the novel macrocycles of the present invention (See Scheme III). In one embodiment (bold-face numerals corresond to compound numerals in Scheme III), a foundation was laid by anchoring to solid support an orthogonally protected trifunctional compound such as N-Fmoc-S-trityl-L-cysteine to give 2 (step a).
The cycle was then built up (steps b,c,d) by coupling desired amino acid and ATA building blocks to provide intermediates such as 5. To facilitate cyclization, a bromoacid was installed at the terminal position (step f) to give intermediate 6. Bromoamide 6 was then treated trifluoroacetic acid (TFA) to liberate it from solid support and to unmask its side chain functional groups.
Intramolecular attack of the revealed cysteine sulfhydryl function on the terminal alpha-bromoamide then formed macrocycle 7, a 19-membered bicycle that possesses a number of significant backbone modifications including two thioether linkages and the rigid 2-mercaptonicotinamide unit whose pyridine type nitrogen may accept hydrogen bonds.

_87-Trt o Trt o N
FmocHtQ - a". N'~ bi NH H
FmocHN H
'O -2 FmocH ~'~ ~ ~ Otbu c HFmoc bu .I
O
NH H f ----O -' ~ ~ Otbu H
Scheme III
Method for assembling a specific pseudopeptide macrocycle (shaded spheres represent a solid support):
a) Fmoc-Rink amide MBHA resin, 30% piperidine in DMF, then 5 eq Fmoc-S-trityl-L-cysteine, DIC/HOBT in NMP; b) as for a), but with 5 eq Fmoc-L-tyrosine-O-t-butyl ether; c) as for a), but with 5eq Fmoc-L-Phe[y~CH2S)-2-mercaptonicotinate; d) as for a), but with 5eq Fmoc-D-alanine; e) 30% piperidine in DMF, then 5eq bromoacetic acid and DIC in NMP; f) 38:1:2 TFA:H20:Et3SiH for 2h, lyophilize, wash (ether, 2:x) then 2eq of diisopropylethylamine in 1:1 CH3CN:H20, 24h.

_88_ Macrocyclization kinetics of 7 is accelarated by the addition of a base. LCMS is utilized for monitoring ring closure. The two major natural isotopes of bromine in linear material were plainly visible at 80 and 82 m.u.
(mass units) over the calculated mass of the cycle. Ring closure was accompanied by a shift in HPLC retention time and a change in the mass spectrum to a non-doubled peak consistent with the mass calculated for cyclic product (for 7, calculated MW=501.41, found MH+=502.2). Under the conditions described, LCMS revealed little or no dimer formation during closure of 7 or any of the other macrocycles made regardless of ring size or structural rigidity. HPLC traces of macrocycles bearing racemic ATAs, such as compound 32 as shown in Method B below, often displayed a pair of closely eluting peaks having similar abundance and identical mass spectra. Thus both ring diastereomers formed and were separable by HPLC.
By utilizing the large number of ATA building blocks and the large number of amino acids and amino/acid bearing compounds that are available commercially, large numbers of unique macrocyclic pseudopeptides can be synthesized. Approximately seven hundred macrocycles have been made using automated multiple parallel synthesis and representatives from this set are shown below (numerals shown within ring structures represent the number of macrocycle ring atoms and Bn is benzyl).

__gg_ H ;O
~('~N
'13 S
~O
O
NHy The synthesis method of the present invention allows the construction of a library of about 12- to about 24-membered macrocycles in integral steps. This capability is important for varying molecular size, volume and shape. Each library compound also contains at least two thioether linkages to enhance lipophilicity and protease resistance relative to simple cyclic peptides. The position of one thioether is fixed at the cysteine or penicillamine used for ligation, but the others) can be installed at any internal site depending on the coupling order of the ATA and amino acid compounds. As judged by LCMS, macrocyclization occurred readily with a variety of L- and D-amino acids without regard to sequence within the ring. Macrocycles containing single or multiple aryl backbone segments also formed readily, exemplified by compounds 14, 16, 17 and 19-24 This is notable since aryl backbones will impart structural rigidity to macrocycles.
Even more important, ortho-(20), meta-(21) and para-substituted (22) aryl ring backbones all permitted efficient ring closure. Model building suggests that macrocycle conformational bias can be influenced by the ortho- vs. meta- vs. para-substitution pattern of the ATAs used during construction. This is a highly desirable feature for de novo lead generation where the active shapes) and conformations) are unknown.
The process of the present invention utilized in preparation of a library of the macrocycle of Formula I
above may be carried out in any vessel capable of holding the liquid reaction medium. In one embodiment, the process of the invention can be carried out in containers adaptable to parallel array synthesis. In particular, the macrocycle library of this invention can be formed in a well plate apparatus 1 or 3 as illustrated in Figures 1 and 2, respectively, and as described in greater detail below. Such apparatus provide multiple reaction zones most typically in a two-dimensional array of defined reservoirs, wherein one member of the macrocycle library of this invention is prepared in each reservoir. Thus the diverse macrocycle library of the present invention comprises a plurality of reservoir arrays (e. g. well plates), each reservoir or well containing a library compound of the macrocycle library. Accordingly the library compounds are typically identified by reference to their well plate number a;nd their X column and Y row well plate coordinates.
Following simultaneous preparation of the library member compounds in the reservoir array, the compounds can be transferred in whole or in part to other reservoir arrays (e.g. well plates), to prepare multiple copies of the library apparatus or to subject the library to additional reaction conditions. Copies of the library apparatus (daughter well plates, each comprising a 2-dimensional array of defined reservoirs with each reservoir containing a predetermined member of the library) are useful as replaceable elements in automated assay machines. The apparatus of this invention allows convenient access to a wide variety of structurally related macrocyclic compounds. One preferred reservoir array for use in making and using this invention is a multi-well titer plate, typically a 96-well microtiter plate.
Figure 1 illustrates the top surface of a well plate apparatus (1) of the present invention. The well plate (1) is a plastic plate with 96-wells (depressions) capable of holding liquids for parallel array synthesis.
Individual reaction products are prepared in each well and are labeled by the well ~?late coordinates. For example, the library compound at location (2), is identified by the alpha numeric coordinate, "A6".
Figure 2 illustrates a side view of a modified well plate apparatus (2) for use in preparation of the library of the present invention. Well plate (3) contains wells (4) with a filter (5), and a retaining frit (6), and a liquid reaction medium used un carrying out the process (7). The wells have an outlet at the bottom which is sealed by gasket (8) held in place by a top cover (9) and bottom cover (10) maintained in position by clamps (11).
Such well plates are typically prepared using conventional 96-well plates. A hole is drilled in the bottom of each well in the plates and a porous frit is placed in the bottom of each well. The plate is then placed in the clamp assembly to seal the bottom of the wells.
Synthesis is initiated by adding reagents to their individual wells according to their assigned plate coordinates. The plate is then capped and tumbled to mix the reagents. Following completion of the reaction, the solvent and residual volatile reagents may be evaporated with a Speed-vac. The residual products are redissolved in appropriate liquid solvent and the reaction products analyzed, for example, by thin layer chromatography, mass spectrometry and/or nuclear magnetic resonance spectrometry.
One embodiment of the present invention is an assay kit for the identification of pharmaceutical lead compounds. The assay kit comprises as essential parts, (1) a well plate apparatus (containing one of the tetrahydroquinoline compounds in each of its individual wells), and (2) biological assay materials. The biological assay materials are generally known to be predictive of success for an associated disease state.
Illustrative of biological assay materials useful in the kit of this invention include, but are not intended to be limited to, those for conducting various assays such as:
In vi tro assays Enzymatic inhibition, Receptor-ligand binding, Protein-Protein interaction, Protein-DNA interaction, and the like;
Cell based, functional assays:
Transcriptional regulation, Signal transduction/Second messenger, Viral Infectivity, __g3_ Bacteriocidal/Bacteriostatic, Fungicidal/fungistatic,, and the like; and Add, Incubate, & Read assays:
Scintillation Proximit~~ Assays, Glucan Synthase (GS) inhibition assays, Angiotensin II IPA receptor binding assay, Endothelia converting Enzyme [125I] SPA assay, HIV proteinase [125I] ;;pA enzyme assay, Cholesteryl ester transfer (CETP) [3H] SPA assay, Fluorescence Polarization Assays, Fluorescence Correlation Spectroscopy, Calorimetric biosensors, Ca2+ -EGTA Dyes for Cell-based assays, Receptor Gene Constructs for cell based assays, Luciferase, green fluorescent protein, beta-lactamase, Electrical cell impedance sensor assays, and the like, and the like assays.

Preparation 1 Methods for Constructing ATA compounds Method A
O O

BocHN BocHN
-~

OH OH

NH2 NHAlloc BocHN
OMs ---- BocHN

NHAlloc ~OH

NHAlloc BocH ~H BocHN ~
~~~OH

NHAIIoc O NHFmoc O

N-oc-Fmoc-N-E-Boc-L-Lvs-f~CH2Sl-Glv (27) Referring to the Method A Scheme shown above (bold-face compound numberals correspond to bold-face compound numberals referred to below), in a one liter 3-necked 25 flask fitted with a magnetic stir bar, 22.65 grams (92mmo1) of commercially available N-E-tert-butoxycarbonyl-L-lysine 22A and 11.56 grams of sodium carbonate (110mmo1) were added to 400m1 of water and stirred to give a suspension. Addition of 400m1 dioxane to the suspension produced a clear pale yellow solution on stirring. This solution was chilled on an ice-water bath with stirring, then 13.2 grams (11.72m1, 110mmo1) of allyl chloroformate was added over several hours via a dropping funnel. Stirring was continued overnight without replenishing the ice bath to yield a light slurry.
The reaction mixture was basified with a few ml of 1N NaOH, then washed twice with 800m1 portions of ethyl ._g5-acetate (EtOAc). The aqueous layer was acidified to pH ~3 with 1N sodium bisulfate (N;~HS04) whereupon a heavy slurry formed. The slurry w;~s extracted twice into 800m1 portions of EtOAc, then the organic layers were combined, washed with saturated brine solution, dried over sodium sulfate (Na2S04) and concenl:rated in vacuo to produce 35grams (>100%) of 23A, a viscous pale yellow oil. TLC
analysis (50:50:1 hexanes:Et~OAc:Acetic acid) showed clean product and this was carried to the next step assuming quantitative yield. The crude mass of 23A was dissolved in 150m1 ethylene glycol dirnethyl ether (DME), chilled to -15°C (ice/water/NaCl slurry) with stirring, then 9.3 grams (10.11m1) of N-methylmorpholine (NMM) was added and the solution re-chilled. Isobutyryl chloroformate (12.56 grams, 11.93m1) was then added in one portion to produce a heavy white precipitate and the resulting slurry was stirred vigorously for 10 minutes on the brine/ice bath.
The slurry was filtered through a fritted funnel into a two liter erlenmeyer flask, retentate washed 5 times with 25m1 portions of DME arid thE~ combined filtrate and rinses were re-chilled to -15°C prior to portion-wise addition of a solution 5.22 grams (138mmo1, l.5eq) sodium borohydride (NaBH4) in 150m1 water. Vigorous evolution of gas was observed after each portion of NaBH4 was added and the mixture was stirred for an additional 10 minutes on the ice/brine bath to produce a colorless emulsion.
TLC (3:1 CH2C12:Et0Ac) revea:Led good conversion to the alcohol. The emulsion was made basic with a few ml 1N
NaOH, then extracted three times with 600m1 portions of EtOAc. The organic layers were combined, washed with saturated brine, then dried over Na2S04 and concentrated in vacuo to yield 25 grams (860) of crude product 24A as a pale yellow oil.
The product was purified on a 500m1 silica column packed and eluted with 7:3 C:H2C12:EtOAc to afford 17.5 grams of a nearly colorless ~~il 24A, 60o overall isolated yield for alloc protection and acid reduction steps. In a 250m1 roundbottom flask, a portion of pure 24A (7.65 grams, 24mmo1) was dissolved in 100m1 dry THF under a nitrogen atmosphere, chilled on an ice bath then 5.06m1 triethylamine (3.67 grams, 36mmo1) was added via syringe.
The solution was stirred for a few minutes on ice, then 2.06m1 (3.05 grams, 26mmo1) of methanesulfonyl chloride (MsCl) was added over a 10 minute period. The clear solution immediately turned cloudy on addition of the MsCl and became a heavy white suspension after all of the MsCl had been added. Stirring was continued for 10-15 minutes following MsCl addition when TLC (3:1 CH2C12:EtOAc) indicated complete consumption of starting alcohol. The slurry was filtered, retentate washed 3 times with 100m1 portions of EtOAc, then the combined filtrate and washes were washed once with cold water, once with cold dilute hydrochloric acid, once with saturated sodium bicarbonate (NaHC03) and once with saturated brine before drying over Na2SOg and concentration in vacuo High vacuum drying of the residue produced 9.22 grams (96%) of 25A as a colorless oil.
In a 250m1 flame-dried flask fitted with a magnetic stir bar, 1.28 grams (0.97m1) of mercaptoacetic acid was dissolved in 30m1 of dry dimethylformamide (DMF) under nitrogen atmosphere and to the mercapto acid was added via syringe 6.4m1 (1.5 grams, 2eq) of a 25 weight percent solution of sodium methoxide in methanol (NaOMe/MeOH).
The clear acid solution turned cloudy white during NaOMe/MeOH addition then returned to its clear and colorless appearance within 30 seconds after the addition was complete. 5 grams of mesylate 25A dissolved in 15m1 dry DMF was then added by pipette in one portion to the stirring dibasic acid and the resulting mixture stirred overnight at room temperature under a blanket of nitrogen. After ~15 hours of stirring, the mixture turned to a heavy slurry with a gray-pinkish color. TLC (50:50:1 ._g7-hexanes:EtOAc:acetic acid) ahowed good conversion, so most DMF was removed with a rotovap at 45°C bath temperature to yield a grayish-white paste. The paste was dissolved in 200m1 water wii:h a few ml of 1N NaOH, and the resulting turbid solution was washed 3 times with 150m1 portions of ethyl ethE~r. The partially clarified aqueous layer was then acidified to pH ~3 with NaHS04 to yield a heavy white slurry that was extracted 3 times with 200m1 portions of EtOAc. The combined EtOAc extracts were washed with saturated brine, dried over Na2S04, then concentrated to afford 4.9 crrams of 26 (98%) as an oil.
The crude product looked relatively clean via TLC
(50:50:1 hexanes:EtOAc:acetic acid) and was carried on without purification.
To 5.1 grams of 26 under nitrogen in a 250m1 flame-dried flask fitted with stir bar was added 60m1 dzy THF
via cannula and the mixture stirred to give a clear and colorless solution. The septum was then removed and in succession were added 3.36 grams solid 5,5-dimethyl-1,3-cyclohexanedione (dimedone), 787 milligrams (0.25eq)of solid triphenylphosphine (PPh3) and 693 milligrams (0.05eq) of solid tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4) to produce a clear orange solution. The flask was recapped, covered with aluminum foil and flushed with nitrogen while stirring at room temperature. TLC in 50:50:1 hexanes:EtOAc:acetic acid showed complete conversion of starting material to a baseline spot after about 1 hour of stirring. Solvent was removed with a rotovap to give an orange oil that was then redissolved in 50m1 EtOAc and extracted 4 times with 25m1 portions of water. To the combined aqueous layers was t:hen added 2 grams solid sodium carbonate and 100m1 of. dioxane.
The resulting light emu7.sion was chilled on an ice bath and when the temperature' of the solution was below 10°C, 4.46 grams of solid fluorenylmethyloxycarbonyl chloride (Fmoc-Cl) was added portion-wise over a one hour period with stirring. The reaction mixture was then stirred overnight without replenishing the ice bath and TLC indicated good conversion of starting baseline material. Next added a few drops of 1N NaOH to the mixture and washed 3 times with 200m1 portions of ethyl ether. The aqueous layer was then acidified to pH ~2 with 1N NaHS04 and the off-white slurry was extracted 3 times with 150m1 portions of EtOAc. The EtOAc layers were combined, washed with saturated brine, dried over Na2S04, then concentrated in vacuo to provide a yellow oil contaminated with dimedone. The oil was applied to a 500m1 silica gel column packed with 50:50:1 hexanes:EtOAc:acetic acid and then eluted with the same solvent. Appropriate fractions were pooled, concentrated, then co-evaporated 3 times with toluene/EtOAc to azeotropically remove excess acetic acid. The resulting colorless oil was extensively dried under high vacuum to yield 2.2 grams (35%) of pure 27.
Method B
OH ~OH OMs -' ' J
H Boc Boc 28 2g 30 32 ~ ~ ~ 1 moc O OH Boy O OH
Referring to the Method B Scheme shown above (bold-face compound numerals correspond to bold-face compound numerals referred to below), in a one liter 3-necked flask, 11.51 grams (100mmo1) of ~3-piperidinemethanol (28) and 10.6 grams (100mmo1) sodium carbonate were _99_ dissolved in 200m1 of water, then 200m1 of dioxane was added and the mixture stirred to produce a pale yellow light emulsion. To the stirring emulsion was next added 24 grams (110mmo1) of di-tert-butyl-dicarbonate and stirring continued overnight. TLC (75:25 CH2C12:EtOAc) revealed complete conversion, so the light yellow slurry was stripped of dioxane, then extracted twice with 300m1 portions of EtOAc. The organic layers were combined, washed with saturated brine, dried over Na2S04, then concentrated in vacuo to yield 20.6 grams (95~) of clean 29 as an off-white solid.
To 20 grams (93mmo1) of 29 under nitrogen in a flame-dried one liter flask equipped with a stir bar was added 400m1 dry THF via cannula. The resulting pale yellow-tan solution was chilled on an ice-water bath, then 14.12 grams (19.44m1, 1:39mmo1) of triethylamine was added by pipette and the solution stirred for a few minutes under nitrogen. Next, 11.72 grams (7.92m1, 102mmo1) of methanesulfonyl chloride was added by syringe over a 10 minute period with vigorous stirring. The clear solution turned to a cloudy whitish-tan suspension immediately upon mesylate addition and became a heavy slurry within a few minutes after all methanesulfonyl chloride had been added. TLC (3:1 CH2C12:EtOAc) revealed complete conversion of the product within 10 minutes of final sulfonyl chloride addition. The slurry was filtered through a fritted funnel and the retentate rinsed 4 times with 100m1 portions of EtOAc. The combined filtrate and rinses were then washed once each with cold water, cold dilute hydrochloric acid, saturated NaHC03, and saturated brine. The organic layer was dried over Na2S04 and concentrated in vacuo to provide 26.94 grams (980) of 30 as a pale yellow crystalline solid. , 2-Mercaptonicotinic acid (11 grams, 7lmmol) was dissolved under nitrogen in 400m1 dry DMF under nitrogen in a one liter 3-necked flask. fitted with a large egg-shaped stir bar to give a bright yellow solution. Next, 7.67 grams (32.5m1, 142mmo1) of a 25 weight per cent solution of NaOMe in MeOH was added to the stirring mercaptonicotinic acid/DMF solution, then 20.8 grams (71mmo1) of 30 dissolved in DMF (52m1 final volume) was added by syringe pump over a two day period. The resulting deep yellow viscous slurry was stripped of DMF, then redissolved in 400m1 of water. The turbid yellow aqueous solution was partially clarified by 3 washes with 300m1 portions of ethyl ether. The partially clarified aqueous layer was acidified to pH ~2 with 1N NaH504 to produce a heavy yellow slurry that was then extracted 3 times with 300m1 portions of EtOAc. The organic layers were combined, washed with saturated brine, dried over Na2S04, and concentrated in va uo to provide 24.6 grams (98%) of crude 31 as a canary yellow solid.
The crude product was recrystallized from neat EtOAc to provide two crops of pale yellow crystals weighing 20.95 grams (83o isolated yield). Purified acid 31 (19 grams, 54mmo1) was combined with 108m1 of a 4N
hydrochloric acid solution in dioxane and stirred in a one liter flask to produce a heavy white-caked precipitate. 100m1 of dioxane was added to loosen the slurry and after one hour of stirring, TLC (50:50:1 hexanes:EtOAc:acetic acid) indicated complete conversion of starting material to a baseline product. A large part of the HC1 was removed under light vacuum on a rotovap, then the remains were neutralized with 1N NaOH solution.
To the neutralized solution was then added with stirring 250m1 of water, 5.72 grams (54mmo1) of solid sodium carbonate and another 100m1 of dioxane. The resulting emulsion was chilled on an ice-water bath, then 19.12 grams (56.7mmo1) of fluorenylmethyloxycarbonyl N-hydroxysuccinimide (Fmoc-OSu) was added portion-wise over a 10 minute period. Stirring was continued for 36 hours to produce an ivory slurry. Dioxane was removed in vacuo, -1.01-then the mixture was washed 3 times with 300m1 portions of ethyl ether. The aqueous layer was then acidified to pH ~2 with 1N NaHS04 and the resulting heavy slurry extracted 3 times with 300m1 portions of EtOAc. The combined EtOAc layers were w~~shed with saturated brine, dried over Na2S04, then concentrated and dried under high vacuum to yield 23.54 grams (92%) of 32 as pale yellow solid that was judged clean by TLC in 50:50:1 hexanes:EtOAc:acetic acid.
Method C
O
Bn0 OOH --- BnO~OH ---~ BnO~OMs oc NHB , NH Boc Me Me BnO~S OH OH
'-- BnO~S
NHFmoc O NHBoc O

N-a-Fmoc-O-benzvl-L-Ser-ft-CH~S1-DL-Alanine !37>
Referring to the Method C Scheme shown above (bold-face compound numerals correspond to bold-face compound numerals referred to below), in a 500m1 round-bottomed flask fitted with a magnetic stir bar, 25 grams (84.6 mmol) of N-oc-Bac-O-benzyl-L-Serine (33) (Novabiochem, San Diego, CA, USA) was dissolved in 150 ml DME and chilled to -15°C with an ice/water/brine bath. 10.23 ml N-methylmorpholine (93 mmol, 1.1 eq) was added to the chilled solution, stirred a few minutes, then 12 ml of isobutyl chloroformate (93 mmol, 1.1 eq) was added in one portion at which time the solution quickly turned to a heavy white slurry. The slurry was stirred vigorously for 10 minutes on the brine/i~e bath, then filtered through a fritted funnel into a two liter erlenmeyer flask. The retentate was washed 5 times with 15 ml portions of DME and the combined filtrate and rinses were re-chilled to -15°C prior to portion-wise addition of a solution 5.3 grams (140mmo1, 1.66eq) sodium borohydride (NaBH4) in 100 ml water. Vigorous evolution of gas was observed after each portion of NaBH4 was added. The mixture was stirred for an additional 10 minutes on the ice/brine bath to produce a colorless emulsion. TLC (3:1 CH2C12:EtOAc) revealed good conversion to the alcohol.
Reaction was quenched with 2000 ml of water and the resulting emulsion was made basic with a few ml 1N NaOH, then extracted three times with 300m1 portions of EtOAc.
The organic layers were combined, washed with saturated brine, then dried over Na2S04 and concentrated in vacuo to yield a pale yellow oil. Product recrystallized from 400 ml of 7:1 petroleum ether: diethyl ether to give 20.28 grams (85%) of alcohol 34 as white needles after washing with hexanes and drying under high vacuum.
In a 1000m1 round-bottomed flask, a portion of pure 34 (9.0 grams, 32 mmol) was dissolved in 125 ml dry THF
under a nitrogen atmosphere, chilled on an ice bath, then 6.69 ml triethylamine (4.86 grams, 48 mmol) was added via syringe. The solution was stirred for a few minutes on ice, then 2.73 ml (4.03 grams, 35.2 mmol) of methanesulfonyl chloride (MsCl) was added over a 10 minute period. The clear solution immediately turned cloudy on addition of the MsCl and became a heavy white suspension after all of the MsCl had been added. Stirring was continued for 10-15 minutes following MsCI addition when TLC (3:1 CH2C12:Et0Ac) indicated complete consumption of starting alcohol. The slurry was filtered, retentate washed 3 times with 100 ml portions of EtOAc, then the combined filtrate and washes were washed once with cold water, once with cold dilute hydrochloric acid, once with saturated sodium bicarbonate (NaHC03) and once with saturated brine before drying over Na2S04 and concentration in vacuo. High vacuum drying of the residue produced 12.7 grams (>100%) of 35 as a colorless oil.
In a 250m1 flame-dried flask fitted with a magnetic stir bar, 1.91 grams (1.6 ml) of DL-thiolactic acid was dissolved in 40 ml of dry di:methylformamide (DMF) under nitrogen atmosphere and to t:he mercapto acid was added via syringe 8.24 ml (1.94 gr,~ms, 2eq) of a 25 weight percent solution of sodium m~~thoxide in methanol (NaOMe/MeOH). About 5.4 grams of mesylate 35 (~15 mmol) dissolved in 35 ml dry DMF w~~s then added by pipette in one portion to the stirring dibasic acid and the resulting mixture stirred ovc=rnight at room temperature under a blanket of nitrogen. After ~15 hours of stirring, the mixture turned to a heav~r slurry with a grayish color. TLC (50:50:1 hexanes:EtOAc:acetic acid) showed good conversion, so most DMF was removed with a rotovap at 45°C bath temperature to ~~ield a grayish-white paste.
The paste was dissolved in 200m1 water with a few ml of 1N NaOH, and the resulting turbid solution was washed 3 times with 150m1 portions of ethyl ether. The partially clarified aqueous layer was then acidified to pH ~3 with NaHS04 to yield a heavy whites slurry that was extracted 3 times with 200m1 portions of EtOAc. The combined EtOAc extracts were washed with saturated brine, dried over Na2SOq, then concentrated to afford 5.6 grams of 36 (101%) as an oil. The crude product looked relatively clean via TLC (50:50:1 hexanes:EtOAc:acetic acid) and was carried on without purification. 3.8 grams of 36 (10.4 mmol) in a 1000 ml round bottomed-flask was chilled on an ice-water bath and combined with 100 ml of a 4N hydrochloric acid solution in ethyl acetate. TLC (50:50:1 hexanes:EtOAc:acet:ic acid) indicated complete conversion of starting material to a baseline product within one hour of HC1 addition. Most of the HC1 was removed under vacuum on a rotovap, then the remains were dissolved in 100 ml of water. To this solution was then added with stirring 4.4 grams (42 mmol, 4 eq) of solid sodium carbonate and then 75 ml of dioxane. The resulting emulsion was chilled on an ice-water bath, then 3.86 grams (11.4 mmol) of fluorenylmethyloxycarbonyl N-hydroxysuccinimide (Fmoc-OSu) was added portion-wise over a 10 minute period. Stirring was continued overnight and TLC (50:50:1 hexanes:EtOAc:acetic acid) of the resulting ivory slurry showed complete consumption of starting 36.
Dioxane was removed in vacuo, the mixture was diluted with 200 ml and then washed 3 times with 200m1 portions of ethyl ether. The aqueous layer was then acidified to pH ~2 with 1N NaHS04 and the resulting slurry extracted 3 times with 100m1 portions of EtOAc. The combined EtOAc layers were washed with saturated brine, dried over Na2S04, then concentrated and dried under high vacuum to yield 5.6 grams (110%) of 37 as a colorless oil.
Recrystallization from acetone-hexanes failed, but TLC
(50:50:1 hexanes:EtOAc:acetic acid) indicated ~80%
purity.

-_L05-Prebaration 2 The following ATA compounds have been prepared in accordance with the general methods of Scheme II and can be utilized to form the macrocycle library of the present invention.
O OH N/
OH \
/ S OH ~'' I \ S
\ I NHFmoc NHFmoc O / NHFmoc O OH
FmocH
/ iN
/S OH \ I N
S S OH
NHFmoc O / p OH NHFmoc O O
~~~ ~ OH ~~
~S~OH ~S~ S~~~OH
NHFmoc " NHFmoc HFm~c ~ NH~II~c / /
O
~ ~ \ I S \ I OH
S' Y 'OH S
O
NHFmoc NHBoc NHFmoc N
O OH Fmoc N/
O
BocH OH ~ ~ ~ S \
S :> Y 'OH
NHFmoc NHFmoc O WHFmoc NHBoc O OH
NHBoc / I NHFmoc S OH
Bn~S OH \ \S \
S O
NHFmoc O I / NHI=moc / ~ I \
O OH \ /
OH ~ OH
_S ~ wS
NHFmoc O NHFmoc p BocH OH
NHFmoc O
OH
S
NHFmoc O
O OH
'S
O NHFmoc O
O
OH
CbzH OH
S
NHBoc O
H
OH
~S
O NHFmoc O
0 S~OH O OH
S
NHFmoc O , NHFmoc O

-~_07-OH
~S OH
S
Fmoc O
NHFmoc O
O
~OH
OH
'S OH
S
Fmoc O
NHFmoc O
O
~S OH S OH
NHFmoc O NHFmoc O
S~ OH
F
H moc O
~S OH S OH
NHFmoc , NHFmoc p O
~S ~ ~OH
NHFmoc NHBoc O
~S ~ ~OH
NHFmoc NHBoc _ OH
S ~ ~S
NHFmoc O
OH
~S
NHFmoc O
OH
~S ~ ~S ~ S OH
NHFmoc 0 NHFmoc 0 , O
S OH Si~OH S OH
NHFmoc O , NHFmoc , NHFmoc O
S OH S~ /OH
NHFmoc O , NHFmoc ~O
O'~S~OH
NHFmoc IIO

O~S OH S \
NTHFmoc O NHFmoc O OH
O O
\~~S OH ~S OH
NHFmoc NHBoc , NHFmoc NHBoc O
/~S OH
NHFmoc NHBoc O OH
S \
\ ~' ~S
FmocN / ~ /- NHFmoc O' ~OH
O
J
OH ~ S
FmocN NHI=moc O OH
N/ O
s 'S OH
Fmoc O OH F~nocN
\ ~ OH ~O S \
\~S
NHFmoc NHFmoc O O OH

'OH
I ~II(S
O
N
Fmoc S \ S \
NHFmoc NHFmoc O OH O OH
N~
~S
F oc O ~ ~OH
S \
/OH
I ~S
NHFmoc O ~ moc O OH ' H
\ OH
\ S
J
Fmoc O
O OH , Fmoc -:L11-N~N~N O
S \ OH ~ N\ ~
S~ v 'OH
NHFmoc O
NHFmoc O O
S~~~OH
~S = OH
NJ ~ Fmoc NHAlloc Fmoc N~N~N O
~--~N\ ~
S- v 'OH
NHFmoc Fmoc H
FmocHl OH

FmocHN
and NHBoc FmocHN S OH
/ S \ O
\ ~ ~ /

Example 1 Method for Preparation of Libraries of Macrocvclic Pseudopeptides and Oraanopeptides Trt O
Trt O N
FmocHN~ ~ N'~~ _~ NH H
FmocHN H
'O -6 FmocH ~~ ~ ~ Otbu c Trt p N'HFmoc N
mr~
~~u H
of H
Cvclo - (Cvs-Tvr-2-Nic fy~H,~.,;;~ -Phe-D-Ala-a-acetate-CH Sl)-(11) 100mg (50micromoles) of Fmoc-Rink-Amide AM or Fmoc-Rink-Amide MBHA resin was placed in a polypropylene reaction vessel (one of 36 such vessels contained in the reaction block of an Advanced Chemtech Model 357 multiple peptide synthesizer). Referring to Scheme III above (bold-face compound numerals and lettered steps correspond to bold-face numerals and lettered steps referred to below), steps (a) through (e) were performed robotically at room temperature according to a custom program. The resin was then washed twice with 1.5m1 of N-methyl-pyrrolidinone (NMP). In Step (a), the Fmoc protecting group was removed by agitating the resin with 1 ml of 30o piperidine in dimethylformamide (DMF) for 3 minutes. The resin was drained and a second 1ml portion of piperidine/DMF solution was added, agitated for 10 minutes, then drained. The resin was then washed and drained 5 to 6 times with 1.5m1 portions of NMP. To the washed resin was then added 300uL of NMP followed by 1m1 of a 0.5M solution of N-Fmoc-S-trityl-L-Cysteine (5 equivalents of protected amino acid to resin-bound amine) and 0.5M hydroxybenzotriazole (HOBT) in NMP and 250u1 of a 1.OM solution of diisopropylcarbodiimide (DIC) in NMP.
This mixture was then agitated for 45 min., drained and the resin rinsed 4 times with 1m1 portions of NMp.
The coupling and rinse steps were repeated one time to yield resin-bound intermediate 6. Single coupling was also acceptable in many cases. Intermediate 6 was then subjected piperidine, NMP washes and coupling as for step (a), but with a 0.5M solution of N-Fmoc-L-Tyrosine-O-tert-butyl ether and 0.5M HOBT in NMP instead of the protected cysteine derivative to produce resin-bound intermediate 7. In step (c), 7 was subjected to the piperidine, washes and coupling regimen of steps (a) and (b), but with a 0.5M solution of N-Fmoc-L-phenylalanyl-[~lCH2S]-2-mercaptonicotinate and 0.5M HOBT in NMP to give compound 8 Compound 9 was produced in step (d) by subjecting compound 8 to the piperidine wash and coupling conditions outlined in step (a) but with 0.5M N-Fmoc-D-Alanine and 0.5M HOBT in NMP. Terminal alpha-bromoamide 10 was made in step (e) by treating compound 9 with piperidine/DMF, washing as in step (a), and coupling with a 0.5M solution of Oc-bromoacetic acid in NMF~ without added HOBT. HOBT is sufficiently nucleophilic to displace the bromine and should thus be omitted when coupling bromoacids.
After coupling of the terminal bromoacid, the resin was washed 4 times with 1.5m1 portions of NMP, then 5 times with 1.5m1 portions of methanol. After methanol washes were complete, resin containing attached compound 10 was dried at room temperature under high vacuum for at least 3 hours. The dried resin was transferred to a screw-capped vial and cleavage effected by agitating the resin with 2.5m1 cleavage cocktail (38:1:1 trifluoroacetic acid(TFA):H2O:triethylsilane (Et3SiH) or 19:1 TFA:H20) for 1.5 hours ~~t room temperature. Another 2 ml of cleavage cocktail wa;~ added and agitated for 1.5 hours, then the entire mixture was filtered through a porous plastic frit. The retentate was washed twice with 2.5 ml portions of water and the combined filtrate and washes were frozen and lyophilized to yield a yellow-orange amorphous solid. This solid was mixed vigorously with 20m1 diethyl ether, chilled to -20°C and centrifuged at 15K rpm, 4°C for 10-15 minutes, then the ether was decanted. The remaining off--white pellet subjected to another ether wash and centr~_fugation run. Ether was decanted and the pellet allowed to air dry, at which time the white to off-white solid was dissolved in 10m1 1:1 CH3CN:H20, filtered through a 0.45uM teflon filter disc, then lyophilized from a tareci scintillation vial. The crude material was then cycli.zed by dissolution in 10m1 1:1 CH3CN:H20 plus 100uL of 1M DIPEA in CH3CN.
Other bases that promotE~d efficient cyclization were 2,6-lutidine and proton sponcre ([1,8-Bis(dimethylamino)naphthalene]). Triethylamine, pyridine and tetramethylguanidine were sufficiently nucleophilic to displace the bromine in preference to promoting cyclization. Cyclization was allowed to proceed for 24 hours at room temperature. LCMS analysis showed a single major peak (purity~80o) having an MH+ of 502.2, consistent with the calculated mass far 11 of 501.4.
The abbreviations "Cys" and "BrA" represent cysteine and bromoacetic acid residues, respectively, and these two residues define the ligation point for ring closure.
Although the majority of cycles synthesized to date use L-Cysteine for ring closure, a number of other cycles have been successfully constructed with D-Cysteine. In addition, it is anticipated that other trifunctional compounds bearing an amine, an acid and a mercapto function (for example, penicillamine), are likely to function in the scheme as does the cysteine residue.
Also, other bromoacids will likely function in place of bromoacetic acid.
Macrocycles 38 and 39 below have been constructed and demonstrate ATA compatibility with commercially available amino acids. Further, macrocycle 38 shows that ATAs provide easy access to novel macrocycles containing aryl backbone elements.

H
O

a S

NH 'Nh~
~O
O
OH

Example 2 Methods for Construc:_tina Tailed Macrocvclic Pseudopeotides and Oraanopeotides According to a general method for constructing resins bearing both free amine groups) and an acid labile carbamate linkage, described and shown below, 11 grams (8.58 millimoles) of p-nitrophenyl carbonate Wang resin (~~pNP-Wang" a commercially available material from Novabiochem, Catalog# 01-64-0123) was swollen with 75m1 of 1:1 dichloromethane (DCM or CH2C12:N-methylpyrrolidinone (NMP), then combined with a 100m1 solution containing 429mmo1s (5 equivalents) of the diamine of choice. The resulting bright yellow slurry is then agitated on a peptide shaker, rotor or other mixing device for 24-48 hours at room temperature. The slurry is next drained using a fritted funnel or peptide shaker flask, then washed with 200m1 each of DCM, NMP and methanol. The wash cycle (DCM, NMP
and methanol) is repeated i:wice more, then the resin is dried under high vacuum for several days to provide the desired product in quantitative yield based upon mass change.
By this method was prepared piperazine-N-carbamoyl-Wang (resin P) (loading: 0.81mmo1/gram of resin) as well as both p- and m-xylenediamine-N-carbamoyl-Wang resins (resins PX and MX, respectively at 0.78mmo1/gram) in multigram quantities.
Alternatively, the desired diamine (or higher order amine) can be similarly attached to resin in situ by mixing at least a five-fold molar excess of di- or higher order amine with pNP-Wang resin in 1:1 DCM:NMP
for 16-48 hours at room temperature. After washing as above, the resin contains at least one free amino function ready for elaboration and at least one acid labile carbamoylated amine function as the link to resin.

-~119-r s a x s ~ c 5 c w > w ~i O Q Q Q
a Q e: o' c c O ~ ~ ~ -v '° 3 w a a. 3 3 O a ~ W i4 S. p 'a.
4i E ~ E~
d ~_ ~ p _Of ul ci X w x U w Z a eo - p. av ~Z amz Q
d aii 5d c~ -wc - _ac co ~c :°. ~« ~E
and ~ ~ c c : ~ d v GC C'~ c: c v tEo C aft G ~ a X m p es 'o Z 8 d y X ' ao : "' 3 oC c~ a E '~_ c G. o h a C
d a oVr m a-° ~r.e n a N o~~ E
w_C o ~
E ~ a to \ /
dd o r~ d NO N o / ~ 0 0 0 '~° E

v' / /
o v o-~~ 3 o z o 0 0 0 O O O
z~ a zs z \ //
z = \ / \

x z z n n x x SUBSTITUTE SHEET (RULE 26) N-r~-xvlenediamino-Cvclo-(Cys-Phe-Glv[t(1CH?S1-Phe-Arg-a acetate-[~rCHZSJ? (18, Scheme IV) w N ~' U
s < < m m <
\ zx \ g a o I o I /
O / Z 'GO :.
N . ~
, H ~. I~ H
O
zx x z ~o v / O ~~~ vi < a x N ~ a z~ xz_ ,_z o =-~!~ n a _~ \ <
I\
I / m m H E
d ~ a w t vd ~ U
ewg ~ \ o a I \ ~
I / .- z o / ~ a m = o m N //
_ a z z ,.
J

z x xz o ."~ a.
U y C ~ ~ ~<n v a cn '_ ~ a /z o U c_si \ a z I /
o a t z ~ w m z a \

.,.
e.
r O OD ~ U
Q ~ O _~o __ Z
\ _ _ /
SUBSTITUTE SHEET (RULE 26) WO 98/46631 - ~ 2~~~ -_ PCT/US98/07135 Steps: a) PX-Wang resin, then 5 eq Fmoc-S-trityl-L-cysteine, DIC/HOBT in NMP, wash; b) 30% piperidine in DMF, wash, then 5 eq Fmoc-L-phenylalanine; c) as for b), but with 5 eq of Fmoc-L-Phe[yrCH2S]-Gly-OH; d) as for b), but with 5 eq of Fmoc-L-Arg(Pmc)-OH where Pmc is defined below; e) as for b), but with 5 eq bromoacetic acid and DIC in NMP without HOST; f) 38:1:1 TFA:H20:Et3SiH for 24 hr., lyophilize, then 2 eq of diisopropylethylamine in 1:1 CH3CN:H20.
With respect tc Scheme IV (bold-face compound numerals and lettered step~~ correspond to bold-face compound numerals and lettered steps referred to below), 65mg (50micromoles) of PX resin was placed in a polypropylene reaction ves~;el Cone of 36 such vessels contained in the reaction flock of an Advanced Chemtech Model 357 multiple peptide synthesizer). Referring to Scheme IV, steps (a) through (e) were performed robotically at room temperature according to a cumtom program written by the bench chemist. The resin was SUBSTITUTE SHEET (RULE 26) washed and drained twice with 1.5m1 of N-methyl-pyrrolidinone (NMP). In Step (a), to washed resin was then added 300uL of NMP followed by 1m1 of a 0.25M
solution of N-Fmoc-S-trityl-L-Cysteine (5 equivalents of protected amino acid to resin-bound amine) and 0.25M
hydroxybenzotriazole (HOBT) in NMP and 250u1 of a 1. OM
solution of diisopropylcarbodiimide (DIC) in NMP. This mixture was then agitated for 45 min., drained and the resin rinsed 4 times with 1m1 portions of NMP. The coupling and rinse steps were repeated one time to yield resin-bound intermediate 13. Single coupling was also acceptable in many cases. In Step (b), the Fmoc protecting group of 13 was removed by agitating resin with 1 ml of 30% piperidine in dimethylformamide (DMF) for 3 minutes. The resin was drained and a second 1m1 portion of piperidine/DMF solution was added, agitated for 10 minutes, then drained. Resin was then washed and drained 5 to 6 times with 2.5m1 portions of NMP.
To washed resin was then added in succession 300uL of NMP, 1m1 of a solution 0.25M each in N-Fmoc-L-Phenylalanine and HOBT and 250u1 of a 1.OM solution of diisopropylcarbodiimide (DIC) in NMP. This mixture was then agitated for 45 min., drained and the resin rinsed 4 times with 1m1 portions of NMP. The coupling and rinse steps were repeated one time to yield resin-bound intermediate 14. In step (c), 14 was subjected to piperidine, washing and coupling regimen of step (b), but instead with N-Fmoc-L-phenylalanyl-[ylCH2S]-glycine to give 15. Compound 16 was produced in step d by subjecting 15 to the piperidine, wash and coupling conditions outlined in step (b) but with N-Fmoc-L-Arginine(Pmc) as the amino acid [where Pmc=2,2,5,7,8-Pentamethylchroman-6-sulfonyl, an acid labile arginine side chain protection function]. Terminal alpha-bromoamide 17 was made in step (e) by treating 16 with piperidine/DMF, washing as in step (b), and coupling with a 0.25M solution of alpha-bromoacetic acid in NMP
without added HOBT. HOBT is sufficiently nucleophilic to displace the bromine and should thus be omitted when coupling bromoacids in this step. After coupling of the terminal bromoacid, the resin was washed 4 times with l.5ml portions of NMP, then 5 times with l.5ml portions of methanol. After methanol washes were complete, resin containing attached 17 was dried at room temperature under high vacuum for at least 3 hours. The dried resin was transferred to a screw-capped vial and cleavage effected by agitating the resin with 2.5m1 cleavage cocktail (38:1:2 trifluoroacetic acid(TFA):H20:
triethylsilane (Et3SiH) or 19:1 TFA:H20) for 1.5 hours at room temperature. Anoi:her 2 ml of cleavage cocktail was added and agitated for 1.5 hours, then the entire mixture was filtered through a porous plastic frit. The retentate was washed twices with 2.5 ml portions of water and the combined fi=Ltrate and washes were frozen and lyophilized to yield ~~ yellow amorphous solid.
Workup and cyclization were performed as in Example 1 above. Cyclization was a~_lowed to proceed for 24 hours at room temperature. LCMS analysis showed a single major peak (purity~80%) having an MH+ of 790.4 and an [M+2H]2+ of 395.8, consistent with the calculated mass for 18 of 789.
The bottom part of the scheme depicts a generic form of the synthetic method u~;ed to construct macrocycles such as 18."Cys" and "BrA" represent the cysteine and bromoacetic acid residues, respectively and these two residues define the ligation point for ring closure.
Though the majority of cycles synthesized used L-Cysteine for "Cys", a number of cycles were successfully constructed with D-Cysteine at the "Cys"
position.

Example 3 Following the procedures of Example 1 above, the following library compounds have been prepared. The number within the ring specifies the number of ring atoms.
S O
HN 12 ' NH2 NH
S B n S ~~O
O
-'S
HN O

S
O

-1.24-HN
,, 1 ~3 ~N

S
O

U
Met R ~ H N~ R~=
S H
15 °
Me NH NHt ~
~ Me' S-r \\
O M ,"
/ Me \ H~~'~a~~~~"
O Me ~.
H H x~"
S
15 ° H
HN °
O
M~''~~, NH N~
M,~~
S O H C
Me I
w."
NHZ \ ~,"

HO /

WO 98/46631 PCT/iJS98/07135 -:126-O
O R'~ N~ R~ H
p ,~-r' N
" 16 0 O

~O
O
O ~ ~ ; H O
N
S
O S ~ S
16 o S 16 0 ~N V N
,NH NH2 NH NHZ
N
O H r';~' O N-T' R2 O H ~~' O

O Me R~_ H2N\/\/H / xIIII
y"~ ~ W~ ~ ~S'y~ ~~S'H'~ HO~SSyh y"4 HO~
Me HO
Me Me' ' S '~ Met HO
HO~
O
Rr H~\/~\/~'w1 \ ~~b \~°"~~ HO~~'v, O
HO / O HO~~""~ ~
Me NH
Me~SW~v H2N~NW~~~ HO/~~'', H
Me Me' ' \ \ ~~ HO HO~
/
HO

y H O
s S
O
NH N~
o M8 H Nw, Rt- 2 ~..~ I \ ~~.y~ I \ '.y~ ~~~.y~ Hp~~.ys HO~sy, HO
Me Me' ' \ S ~~ HO HO~
I /
Rx HpNw.,~~ I \ ',~Y \ y, ~ Hp~y~., p ~ , '~, ~ /'~a HO / O HO' v "~
NH
H Me~S~~~~~ Hz~'~~N~~~'~ HO/v,''.
H
Me Me' ' \ \ ~~ HO HO~
I~ I/
HO

-~_28-O
N
R' / H
O S, O

~O
~l N
R~_ H2N "~"~ \ /"w, \ ", ~, ""' "~"" Me~"~""", HO ~ ~ Me O Me Me ~ /., ~
HO' v ""'~ "'"w HO Me' ' \
S ~ Met f.~0 HO~
O

WO 98!46631 PCT/US98/07135 HI
Me~."",~
R
Rte. ~-~ ,~' ~
s HN
18 ~ Me 18 g ~ S
O OH
O ~ ~ O
O
R2 N~ R
H

~-- Me' I/ H Me~, O C
Me H 2N ~/\/""'".", M a "" ~ /""
"x '"'' HO- v """~' HO °'' Me ''"i~,"
HO
Met H~ Me HO
Me HpN ~","w" HO ~~~,~"~ HO/"~,~~
U

~~ R~ ~~

-:L30-H N g' Me O

S NH N~
O
H2N O H \
Me OH
R~= H2N h~wh, M
Me hAyAyN, \ ~hM~~Yhi HO / / /
O Me M "' ~ /~
~~b~ ~~~h~h DIY
HO ~ HO
Me ~ HO~
\ 'OI HO~

H O
S
O
N!~
NH
S
i p H ~ ~ ~ OH
O
Me R~- H2N ~~'~~~ M ~ H O
M
b w w4 \ .
/ / H / \
O Me M~/,~,.,~ H
~~~~', Me O
Rz H O Rz H O
O ~'''~ N ~ N
S O
O \~ S O
N
NH NH z Z 1 NH NH z O S
N ~O
t H ~ ~ OH ~ ~ H OH
O
R 2. H zN w~~,~ H O 'ww i"w HO '~
O

Claims (36)

I claim:

1. A library of structurally related macrocyclic compounds of the formula wherein L is a divalent group of the formula 1(CH2)0-1-C6H4CH2 2 or -CHR a- wherein R a is hydrogen or an organic moiety;
R z is hydrogen or methyl;
B is hydroxy, or a group of the formula -NR y R t wherein R y is hydrogen, alkyl, aryl or heterocycle, and R t is hydrogen, a solid support, or R t is a group of the formula wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and R y" is hydrogen or a substituent derived from an electrophilic reagent; or R y and R y" taken together with the atoms to which they are bonded form a 6- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or R y or R y" taken together with -T- and the atoms to which they are commonly bonded form a 4- to 7-membered ring or R y and R y" each taken together with T and the nitrogen atom to which they are respectively bonded form a 5- to 7-membered ring;
Z is a divalent group of the formula a (AA1)a1-(AA2)a2-(ATA1)t1-(AA3)a3-(ATA2)t2-(AA4)a4-(ATA)t3-(AA5)a5-(AA6)a6b wherein each of AA1-6 is independently a divalent group of the formula and each of ATA1-3 is independently a divalent sulfur group of the formula and wherein a1, a2, a3, a4, a5, a6, t1, t2, and t3 are independently 0 or 1, provided that t1 + t2 + t3 - 1, 2 or 3; and provided further that -Z- is selected to provide a 12- to 40-membered macrocycle ring of Formula I;
wherein in the above formulas for AA1-6 and ATA1-3, W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N
and S;
Q and Q' are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;
m is 0, 1 or 2;
R s is hydrogen or an organic moiety; and R n' is hydrogen, or an organic moiety, or R n taken together with Q and the atoms to which they are bonded form a 4- to 7-membered ring;
R n' and R s taken together with -Q'- and the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 22 carbon atoms; or R n' or R s taken together with -Q'- and the atoms to which they are commonly bonded form a 4- to 7-membered ring.

2. The library of claim 1 wherein B is amino.

3. The library of claim 1 wherein B is a group of the formula -NR y Rt wherein R y is hydrogen, alkyl, aryl or heterocycle and R t is hydrogen, a solid support, or R t is a group of the formula

4. The library of claim 3 wherein E is hydrogen.

5. The library of claim 3 wherein E is a substituent derived from an organic electrophilic agent.

6. The library of claim 1 wherein a1 + a2 + a3 + a4 + a5 + a6 - 1, 2 or 3.

7. The library of claim 6 wherein t1 + t2 + t3 = 1 or 2.

8. The library of claim 1 wherein each -AA- is a divalent group derived from a proteiogenic amino acid and isomers thereof.

9. The library of claim 1 wherein W is a divalent organic group selected from the group consisting of -CHR8-, -CR4R5CR6R7-, and a 5- or 6-membered aromatic ring containing 0 to 4 heteroatoms selected from O, N and S;
wherein R8 is hydrogen or an organic group;
R4, R5, and R6 are independently hydrogen or alkyl;
and R7 is hydrogen, hydroxy, protected hydroxy, amino or protected amino or substituted amino wherein the substituent is derived from an electrophilic group.

10. The library of claim 1 wherein Q is a group of the formula wherein R1 is hydrogen, or a non-interfering substituent; and R2 is hydrogen or an organic moiety; or R1 taken together with R n forms a 4- to 7-membered ring, or R1 taken together with Rs forms a 4- to 7-membered ring; or R1 taken together with R2 forms a 3- to 6-membered ring.

11. The library of claim 1 wherein -Z- is selected to provide a 12 to 24 membered macrocycle of Formula I.

12. A library of macrocyclic compounds of the formula wherein L is a divalent group of the formula 1(CH2)0-1-C6H4CH2 2 or -CHR a- wherein R a is hydrogen or an organic moiety;
R z is hydrogen or methyl;
B is hydroxy or a group of the formula -NR y R t wherein R y is hydrogen, alkyl, aryl or heterocycle, and R t is hydrogen, a solid support, or R t is a group of the formula wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and R y" is hydrogen or a substituent derived from an electrophilic reagent; or R y and R y" taken together with the atoms to which they are bonded form a 6- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or R y or R y" taken together with -T- and the atoms to which they are commonly bonded form a 4- to 7-membered ring or R y and R y" each taken together with T and the nitrogen atom to which they are respectively bonded form a 5- to 7-membered ring;
Z is a divalent group of the formula a(AA1)-(ATA1)-(AA3)b wherein AA1 and AA3 are independently a divalent group of the formula and ATA is a divalent sulfur group of the formula wherein W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from the group consisting of O, N and S;
Q and Q' are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;
m is 0, 1 or 2;
R S is hydrogen or an organic moiety; and R n' is hydrogen, or an organic moiety, or R n taken together with Q and the atoms to which they are bonded form a 4- to 7-membered rind;
R n' and R s taken together with -Q'- and the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or R n' or R s taken together with -Q'- and the atoms to which they are commonly bonded form a 4- to 7-membered ring.

13. The compound of claim 12 wherein B is amino.

14. The compound of claim 12 wherein B is a group of the formula -NR y R t wherein R y is hydrogen, alkyl, aryl or heterocycle, and R t is hydrogen, a solid support, or R t is a group of the formula

15. The compound of claim 14 wherein E is hydrogen.

16. The compound of claim 14 wherein E is an organic substituent derived from an electrophile.

17. The compound of claim 12 wherein W is a divalent organic group selected from the group consisting of -CHR8-, -CR4R5CR6R7-, and a 5- or 6-membered aromatic ring containing 0 to 4 heteroatoms selected from O, N and S;
wherein R8 is hydrogen or an organic group;

R4, R5, and R6 are independently hydrogen or alkyl;
and R7 is hydrogen, hydroxy, protected hydroxy, amino or protected amino or substituted amino wherein the substituent is derived from an electrophilic group.

18. The compound of claim 12 wherein AA1 and AA3 are independently a divalent residue of a proteiogenic amino acid or isomer thereof.

19. The compound of claim 12 wherein Q is a group of the formula wherein R1 is hydrogen, an organic moiety or a non-interfering substituent; and R2 is hydrogen or an organic moiety; or R1 taken together with Rn forms a 4- to 7-membered ring, or R1 taken together with R s forms a 4- to 7-membered ring; or R1 taken together with R2 forms a 3- to 6-membered ring.

20. A process for preparing a diverse library of structurally related macrocyclic compounds of the formula wherein L is a divalent group of the formula 1 (CH2)0-1-C6H4CH2 2 or -CHR a- wherein R a is hydrogen or an organic moiety;
R2 is hydrogen or methyl;
B is hydroxy or a group of the formula -NR y R t wherein R y is hydrogen, alkyl, aryl or heterocycle, and R t is hydrogen, a solid support, or R t is a group of the formula wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and R y" is hydrogen or a substituent derived from an electrophilic reagent; or R y and R y" taken together with the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or R y or R y" taken together with -T- and the atoms to which they are commonly bonded form a 5- to 7-membered ring or R y and R y" each taken together with T and the nitrogen atom to which they are respectively bonded form a 5- to 6-membered ring;
Z is a divalent group of the formula a(AA1)a1-(AA2)a2-(ATA1)t1-(AA3)a3-(ATA2)t2-(AA4)a4-(ATA3)t3-(AA5)a5-(AA6)a6b wherein each of AA1-6 is independently a divalent group of the formula and each of ATA1-3 is independently a divalent sulfur group of the formula and wherein a1, a2, a3, a4, a5, a6, t1, t2, and t3 are independently 0 or 1, provided that t1 + t2 + t3 - 1, 2 or 3; and provided further that -Z- is selected to form a 12- to 40-membered macrocycle ring of Formula I;
wherein in the above formulas for AA1-6 and ATA1-3, W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N
and S;
Q and Q' are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;

m is 0, 1 or 2;
RS is hydrogen or an organic moiety; and Rn' is hydrogen, or an organic moiety, or Rn taken together with Q and the atoms to which they are bonded form a 4- to 7-membered ring;
Rn" and Rs taken together with -Q'- and the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or Rn" or Rs taken together with -Q'- and the atoms to which they are commonly bonded form a 4- to 7-membered ring;
wherein said process comprises the steps of (a) covalently bonding an amino-protected, thiol-protected starting material of the formula , wherein RZ is hydrogen or methyl, and PS and P are thiol- and amino-protecting groups, respectively, to a solid support comprising acid reactive groups;
(b) removing the amino protecting group on the solid support-bound starting material;
(c) coupling the resulting solid support-bound thio-protected starting material with an amino-protected amino acid of the formula or an amino-protected thioether acid of the formula wherein in said formulas P is an amino-protecting group and, Rn, Rn', Q, Q', Rs, m and W are as defined above;
(d) removing the amino-protecting group from the coupled solid support-bound product;
(e) optionally repeating step (c) and step (d) one or more times using the same or a different amino-protected amino acid or amino-protected thioether acid (f) coupling the solid support-bound product with an organic acid of the formula wherein L is as defined above and X is a leaving group subject to nucleophilic displacement;
(g) removing the thiol-protecting group to enable macrocycle formation by nucleophilic displacement of the group X and cleaving the covalently bound product from the solid support; and (h) when the acid reactive group on the solid support is a covalently bound group of the formula , optionally reacting the resulting product of Formula I
wherein B is a group of the formula with an amine reactive organic electrophilic agent;
provided that at least one of the steps (c) and (e) is carried out using an amine-protected aminothioether acid.

21. The process of claim 20 wherein the solid support is a Merrifield resin, Wang or Rink amide resin.

22. The process of claim 20 wherein the acid reactive group on the solid support is a covalently bound group of the formula

23. The process of claim 22 wherein the product is reacted with an electrophilic agent having a molecular weight from about 30 to about 600 and selected from the group consisting of aldehydes, organic halides, acyl halides, sulfonic acid esters, organohaloformates, organosulfonyl halides, organic isocyanates, organic isothiocyanates, aldehydes and ketones.

24. The process of claim 20 wherein the thiol-protecting group is removed to enable macrocycle formation by nucleophilic displacement of the group X

before the covalently bound product is cleaved from the solid support.

25. The process of claim 20 wherein the thiol-protecting group is removed to enable macrocycle formation by nucleophilic displacement of the group X
after the covalently bound product is cleaved from the solid support.

26. The process of claim 20 wherein each amino-protected amino acid used in the process is a proteiogenic amino acid or isomer thereof.

27. The process of claim 26 wherein P is 9-fluorenylmethyloxy carbonyl.

28. The process of claim 10 wherein each protected amino acid used in the process has a molecular weight of about 75 to about 700.

29. The process of claim 20 wherein each aminothioether acid used in the process has a molecular weight of about 150 to about 800.

30. The process of claim 20 wherein steps (c) and (d) are repeated again using the same or a different amino-protected amino acid or thioether acid.

31. The process of claim 20 further comprising contacting the product of step (g) in solution with a non-nucleophilic base selected from 2,6-lutidine, diisopropylethylamine or proton sponge.

32. An assay kit for identification of pharmaceutical lead compounds, said kit comprising biological assay materials and a well plate apparatus wherein each well in said apparatus contains a library compound of the library of claim 1.

33. The assay kit of claim 32 wherein the biological materials are selected for performing at least one assay test selected front the group consisting of in vitro assays, cell based functional assays, and add, incubate, and read assays.

34. An apparatus suitable as a replacement element in an automated assay machine as a source of individual members of a library of structurally related compounds, said apparatus comprising a 2-dimensional array of defined reservoirs, each reservoir containing a library compound from said library, wherein said structurally related compounds are of the formula wherein L is a divalent group of the formula 1 (CH2)0-1-C6H4CH2 2 or -CHR a- wherein R a is hydrogen or an organic moiety;
R Z is hydrogen or methyl;
B is hydroxy or a group of the formula -NR y R t wherein R y is hydrogen, alkyl, aryl or heterocycle, and R t is hydrogen, a solid support, or R t is a group of the formula wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and R y" is hydrogen or a substituent derived from an electrophilic reagent; or R y and R y" taken together with the atoms to which they are bonded form a 6- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or R y or R y" taken together with -T- and the atoms to which they are commonly bonded form a 6- to 7-membered ring or R y and R y" each taken together with T and the nitrogen atom to which they are respectively bonded form a 5- to 7- membered ring;
Z is a divalent group of the formula a(AA1)a1-(AA2)a2-(ATA1)t1-(AA3)a3-(ATA2)t2-(AA4)a4-(ATA3)t3-(AA5)a5-(AA6)a6 b wherein each of AA1-6 is independently a divalent group of the formula and each of ATA 1-3 is independently a divalent sulfur group of the formula and wherein a1, a2, a3, a4, a5, a6, t1, t2, and t3 are independently 0 or 1, provided that t1 + t2 + t3 = 1, 2 or 3; and provided further that -Z- is selected to form a 12- to 40-membered macrocycle ring of Formula I;
wherein in the above formulas for AA1-6 and ATA1-3, W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N
and S;
Q and Q' are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from O, N and S;
m is 0, 1 or 2;
R s is hydrogen or an organic moiety; and R n' is hydrogen, or an organic moiety, or R n taken together with Q and the atoms to which they are bonded form a 4- to 7-membered ring;
R n' and R s taken together with -Q'- and the atoms to which they are bonded form a 4- to 7-membered ring or a bicyclic or tricyclic ring comprising 6 to 12 carbon atoms; or R n' or R s taken together with -Q'- and the atoms to which they are commonly bonded form a 4- to 7-membered ring.

35. The apparatus of claim 32 wherein the library compound in each reservoir is prepared in accordance with the process of claim 19 and wherein each reservoir provides one reaction zone.

36. The apparatus of claim 32 wherein the 2-dimensional array of defined reservoirs is a multi-well microtiter plate.