WO1998046631A1 - 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

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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 pharmaceutical industry.

Synthesis of compounds is an expensive and time consuming phase of research and development. Historically, research chemists individually synthesized and analyzed high purity compounds for biological screening 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 considerably 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 (10^ - 10^) 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 synthesis offers the advantage that the solid support-bound products are easily washed free of excess reagent. Solution phase 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 class 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 is 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 . , Biopolv ers 32, 1519-33 (1992); Szewczuk et al., Biochemistry 31, 9132-40 (1992); Schiller et al . , J. Med Chem. , 34 3125-32 (1991); Toniolo, Int. J. Pept.

Prot. Res., 35, 287-300 (1990); Veber et al . , Nature. 280, 512-4 (1979)]

Barker et al . fJ. Med. Chem. 35, 2040-8 (1992)] and McDowell and Gadek fJACS, 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 Ilb-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 " [ψCH2NH] " 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. Peot . Prot. Res. 42, 284-93 (1993); Anwer et al . , Int. J. Peot. Prot. Res., 36, 392-9 (1990); Spatola et al . , Colloσ. INSERM. 174, 2nd Peptide Forum, 45-54 (1989); and Darlak et al . , Pept. Proc. Eur. Pept. Sv p. 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 compounds from one or more aminothioether 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 the central amide linkage, such as, for example, those described by Spatola, A.F., in Chemistry and Biochemistry of Amino Acids, Peptides and Proteins , B. Weinstein, 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 and acid functionality, including both proteiogenic amino acids and non- proteiogenic amino acids, to construct diverse libraries of peptidomi etic 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 .

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- methylpyrrolidinone (NMP) ; Steps b') through d') as for Step a') but with the respective amino acid (AA) or ATA; e') 30% piperidine in DMF, then 5eq bromoacetic acid and DIC in NMP; f) 38:1:1 trifluoroacetic acid (TFA) :H2θ:Et3SiH for 2 hours, lyophilize, the 2 eq of diisopropylethylamine in 1:1 CH3CN:H2θ.

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 readily accessible source of library compounds .

Brief Description of the Drawings

Fig. 1 is a top view of a well plate in accordance with this invention. Fig. 2 is a side view of a well plate apparatus for use in the process of this invention.

Detailed Description 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. "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 peptido imetic 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 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 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 stored without prior and subsequent intentional mixing.

"Simultaneous synthesis" means making of library compounds within one production cycle of a combinatorial method (not making all library 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 substituents" 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, Cι~ Cio alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkoxy, C7-C12 aralkyl, C7-C12 alkaryl, C1-C10 alkylthio, arylthio, aryloxy, arylamino, C3-C10 cycloalkyi, C3-C10 cycloalkenyl, di (C1-C10) -alkylamino, C2-C12 alkoxyalkyl, Ci-Cβ alkylsulfinyl, C1-C10 alkylsulfonyl, arylsulfonyl, aryl, hydroxy, hydroxy (Cχ-Cιo) alkyl, aryloxy (C1-C10 ) alkyl , Cl-Cio alkoxycarbonyl , aryloxycarbonyl , C1-C10 alkanoyloxy, aryloyloxy, substituted alkoxy, fluoroalkyl, nitro, cyano, cyano (C1-C10 ) alkyl, C1-C10 alkanamido, aryloylamido, arylaminosulfonyl, sulfonamido, amidino, carbamido, , carboxy, heterocyclic radical, nitroalkyl, and - (CH2)m-Z- (Ci-Cio 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, C7-C12 aralkyl, C7-C12 alkaryl, Cl-Cio alkylthio, arylthio, aryloxy, arylamino, C3-C10 cycloalkyi, C3-C10 cycloalkenyl, di (Cl-Cio ) -alkylamino, C2-C12 alkoxyalkyl, Ci-Cβ alkylsulfinyl , C1-C10 alkylsulfonyl, arylsulfonyl , aryl, hydroxy, hydroxy (Cι~ Cio) alkyl, aryloxy (Cl-Cio ) alkyl, Ci-Cio alkoxycarbonyl, aryloxycarbonyl , Ci-Cio alkanoyloxy, aryloyloxy, substituted (Ci-Cio) alkoxy, fluoroalkyl, nitro, cyano, cyano (Ci-Cio) alkyl, Ci-Cio alkanamido, aryloylamido, arylaminosulfonyl, sulfonami o, amidino, carbamido, protected amino, protected carboxy, protected amino, carboxy, heterocyclic radical, nitroalkyl, and -(CH2)m~Z- (Cl-Cio alkyl), where m is 1 to 8 and Z is oxygen or sulfur.

The term "amino acid" as 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-butylalanine, phenylglycine, cyclohexylalanine, beta-alanine, 4-aminobutyric acid and other compounds of the general formula:

HN Q COOH

wherein R_n 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 0, N and S; or R_n 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

HN CHRiCOOH

Rn wherein Ri is selected from the group consisting of hydrogen, methyl, isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, sulfhydrylmethyl, 2 (methylthio) ethyl, benzyl, 4-hydroxybenzyl , 3-indolylmethyl, carboxymethyl, 2-carboxyethyl, carbamidomethy1, 2-carbamidoethyl, 4-aminobutyl, 3-guanadinylpropyl and 4-imidazolylmethyl, and R_n is hydrogen, or R_n and Ri 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(0)X wherein X is a leaving group subject to nucleophilic displacement by nucleophiles. Exemplary of the group X is chloro (an acid chloride), -0C(0)Rχ (an anhydride), or -

0R_X (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 -NHR_r, where R_r 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

wherein

L is a divalent group of the formula - (CH₂) ₀-_ι-C₆H₄CH₂- 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 -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and R is hydrogen, a solid support, or Rt is a group of the formula

N E

wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and Ry" 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

⁽AAι⁾al-⁽AA2⁾a2-⁽ATAι⁾ _tl-⁽AA3⁾a3-⁽ATA2⁾t2-⁽AA4⁾a4-

⁽ATA3 ⁾ 3- ⁽ A₅ ⁾ _a5- ⁽AA6 ⁾ a6^~

wherein each of AAi-β is independently a divalent group of the formula

0

-N- -Q-

Rn

and each of ATAχ-3 is independently a divalent sulfur- linked group of the formula

and wherein al, a2, a3 , a4, a5, a6, tl, t2, and t3 are independently 0 or 1, provided that tl + 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 AAi-6 and ATAχ-3, W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from 0, 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 0, 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;

Rn ' 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 Rn ' or R_s 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 embodiment the library compounds are constructed to have 12 to 24 ring atoms, at least two of which are sulfur. The ring atoms are derived from four types of reactants during the solid phase synthesis: 1) a solid suppor -anchored orthogonally protected trifunctional (protected thiol, protected amino, carboxy) compound (viz. cysteine or penicillamine) ; 2) aminoacids, selected from proteiogenic amino acids and non- proteiogenic amino acids; 3) ATAs (a inothioether acids and their corresponding sulfoxide and sulfone derivatives) ; and 4) alpha-halo acids, preferably alpha- bromo acids or carboxy- or carboxymethyl-substituted benzyl bromides. Generally, macrocycle 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 and 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 (AAχ-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 is hydrogen, alkyl, aryl or heterocycle and Rt is hydrogen, a solid support, or Rt is a group of the formula

T N E

wherein T, Ry" and E are as defined above. The compounds where E is hydrogen can be derived from corresponding library compounds wherein E is a solid support, particularly a Wang resin, by trifluoroacetic acid mediated cleavage, or they can 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 precursor compounds) also represent embodiments of the present invention. A general compilation of solid supports (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 Gϋnther 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) :

Wang p-nitrophenylcarbonate resin:

Rink Amide AM resin:

Rink Amide MBHA resin:

Knorr Resin:

The library compounds of Formula I of this invention wherein B is the group -NRyRt, Rt is a group of the formula

-T- -N-

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

HN T N CO—CH₂-C₆H₄-0-CH₂-C₆H-Polymer

R_y R_y" 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

HN T NH

Ky 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-DIAMINOCYCLOHEXANE

1, 3-BIS (AMINOMETHYL) CYCLOHEXANE

D-CYSTINE

L-ARGININE

2,6-DIAMINOPIMELIC ACID

2, 6-DIAMIN0PIMELIC ACID N-EPSILON-CBZ-L-LYSINE N-EPSILON-ACETYL-L-LYSINE N- ( 3 -AMINOPROPYL) CYCLOHEXYLAMINE ETHYLENEDIAMINE-N, N ' -DIACETIC ACID

N, N ' -DIBENZYLETHYLENEDIAMINE PIPERAZINE 2 -METHY PIPERAZINE 2 , 6-DIMETHYLPIPERAZINE 1- ( 2-AMINOETHY ) 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) ETHYLENEDI MINE 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 2-N-BUTYLAMINOETHYLAMINE

1 , 4-DIAMINOCYCLOHEXANE

S- (2 -AMINOETHYL) -L-CYSTEINE

HYDROCHLORIDE

N, N-DIETHYLDIETHYLENETRIAMINE

TERT-BUTYL N-(6-

AMINOHEXYL) CARBAMATE

HYDROCHLORIDE

1 , 4-BIS (AMINOMETHYL) CYCLOHEXANE

1 , 8-DIAMINO-3 , 6-DIOXAOCTANE

N-BENZYLETHYLENEDIAMINE

PIPERAZINE-2-CARBOXYLIC ACID

DIHYDROCHLORIDE

L-CYSTINE DIHYDROXAMATE -ARGININE HYDROXAMATE

HYDROCHLORIDE

DIAMINOBIOTIN

L-LYSINE HYDROXAMATE

HYDROCHLORIDE

L-LYSINAMIDE DIHYDROCHLORIDE

3 -AMINOPYRRO IDINE

DIHYDROCHLORIDE

(lR,2R)-(-)-l,2- DIAMINOCYCLOHEXANE

(lS,2S)-(+)-l,2- DIAMINOCYCLOHEXANE CIS-1, 2 -DIAMINOCYCLOHEXANE TRANS-1 , 2-DIAMINOCYCLOHEXANE L-CYSTINE L-LYSINE

L ( - ) -ALPHA-AMINO-EPSILON- CAPROLACTAM

L-ARGININE MONOHYDROCHLORIDE L-ARGININE MONOHYDROCHLORIDE L-2,4-DIAMINOBUTYRIC ACID DIHYDROCHLORIDE

L-ORNITHINE MONOHYDROCHLORIDE L-ORNITHINE MONOHYDROCHLORIDE DL-LYSINE MONOHYDROCHLORIDE DL-LYSINE MONOHYDROCHLORIDE L- YSINE MONOHYDROCHLORIDE DL-ORNITHINE MONOHYDROCHLORIDE L-LYSINE DIHYDROCHLORIDE DL-HOMOCYSTINE

TRANS-2 , 5-DIMETHYLPIPERAZINE ETHAMBUTOL DIHYDROCHLORIDE

(lS,2S)-(-)-l,2- DIPHENYLETHYLENEDIAMINE

(lR,2R)-(+)-l,2- DIPHENYLETHYLENEDIAMINE DL-CYSTINE 1 , 3 -DIAMINOPENTANE (4S,5S) -4, 5-DI (AMINOMETHYL) -2,2- DIMETHYLDIOXO ANE PIPERAZINE HEXAHYDRATE (S) - (+) -2-METHYLPIPERAZINE (S)-(+)-2-

( MINOMETHYL) PYRROLIDINE Nl-ISOPROPYLDIETHYLENETRIAMINE (R)-(-) -2-METHYLPIPERAZINE N-PHENYL-4-PIPERIDINAMINE 2 -AMINO-6-FLUOROBENZYLAMINE 2 -AMINOBENZYLAMINE 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 ai + a2 + a3 + a4 + as + aβ = 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 ti + 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 ATAi k, a AAl-ATAl-ATA2-AA4 fc, a ATA1-AA3-ATA2 k, a AAl-ATAl-ATA2-AA4-AA5 -, ATA1-AA3-AA4-ATA3 k, ATA1-ATA2-ATA3-AA5 k, a AAl-ATA2-ATA3-AA5-AAg -- a AA3-ATA2-AA4 b, a ATA2-AA4-ATA3-AA5 k, a AAl-ATAl-ATA2 -ATA3 -, a AA4-ATA3-AA5-AA6 b, a AAl-ATAl -, a ATA1-AA3 k, and a ATAl-AA3 -ATA2 -ATA3 - • One preferred embodiment of the invention is a library of macrocyclic compounds of the formula

wherein

L is a divalent group of the formula ^~ (CH₂) ₀-₁-C₆H₄CH₂ ¹ 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 -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and Rt is hydrogen, a solid support, or Rt is a group of the formula

N

Ry,, wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and Ry" 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 4- 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 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

-(AAl) -(ATAl) -(AA3)-

wherein AAi and AA3 are independently a divalent group of the formula

O

N Q c

Rn. 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 0, 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 0, N and S; m is 0, 1 or 2 ;

Rs 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 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

wherein Ri is hydrogen, or a non-interfering substituent; and

R2 is hydrogen or an organic moiety; or Rl taken together with R_n forms a 4- to 7-membered ring, or Ri taken together with R_s forms a 4- to 7- membered ring; or Ri 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 -CHR8-, -CR4R5CR6R7- , and a 5- or 6- membered aromatic ring containing 0 to 4 heteroatoms selected from 0, N and S; wherein R8 is hydrogen or an organic group;

R4, Rs, 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.

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

wherein L, Z, R_z 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

NHP

wherein R_z is defined as above, and P_s 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 material;

(c) coupling the resulting solid support bound thio- protected starting material with an amino-protected amino acid of the formula

R,

or an amino-protected compound of the formula

(O)_r

N Q' CH- W. COOH

R„ R.

wherein in said formulas P is an amino-protecting group and, R_n, Rn ' / Q> Q'> Rs, ™ 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

1

L COOH

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 C IIO— CH2-C6H -O -CH2-C6H4- Polymer

Ry Ry"

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 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, stoichiometrically 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 optimize yield without complicating product isolation. Using solid phase synthesis techniques, effective work-up to remove any excess reagents comprises washing the solid support bound product one or more times with 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

NHP wherein R_z is defined as above, and P_s 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-methoxypheny1-diphenylmethy1 , 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-butvloxvcarbonvl (Boc) , allyloxycarbonyl (Alloc) , 2 -trimethylsilylethoxycarbonyl (Teoc) , biphenylisopropyloxycarbonyl (Bpoc) , nitroveratryloxycarbonyl (Nvoc) ,

( 4-methoxyphenyl ) diphenylmethy1 (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 30% piperidine in dimethylformamide.

The resulting solid support bound thio-protected starting material is then coupled with an amino-protected amino acid of the formula

N Q COOH

R n

or an amino-protected ATA of the formula

(O) m

N CH- w. COOH

R„ R. wherein P is an amino-protecting group and R_n, Rn ' Q, Q ' , R_s, 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- 10 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 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:

1-AMINO-1-CYCLOPENTANECARBOXYLIC ACID

1-AMINO-1-CYCLOHEXANECARBOXYLIC ACID

N-ALPHA-CBZ-L-ARGININE

N-ALPHA-BENZOYL-L-ARGININE

CHLOROACETYL-L-TYROSINE

DL-M-TYROSINE

L-DOPA

4-BROMO-DL-PHENYLALANINE

DL- -FLUOROPHENYLALANINE

DL-4-CHLOROPHENYLALANINE

O-METHYL-L-TYROSINE

0-BENZYL-L-TYROSINE

L-TYROSINE

3-IODO-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 5-METHYL ESTER

L-GLUTAMIC ACID GAMMA-BENZYL ESTER

L-GLUTAMIC ACID

L-ARGININE

L-ALPHA-AMINOADIPIC ACID

2,6-DIAMINOPIMELIC ACID 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

2-IODOHIPPURIC ACID

2-HYDROXYHIPPURIC ACID

Z-L-ASPARTIC ACID

L-ASPARTYL-L-PHENYLALANINE METHYL ESTER

N-CBZ-L-GLUTAMIC ACID

N-ACETYL-L-GLUTAMIC ACID

D -ALPHA-METHYLTYROSINE

N-CARBOBENZYLOXY-2 -METHYLALANINE

2- (METHYLAMINO) ISOBUTYRIC ACID

ALPHA-HYDROXYHIPPURIC ACID

ETHYLENEDIAMINE-DI (O-HYDROXYPHENYLACETIC ACID)

D- ( - ) -P-HYDROXYPHENYLGLYCINE

D-PENICILLAMINE DISULFIDE

D-TERT-LEUCINE

DL-DOPS

DL-VALINE

DL-ISOLEUCINE

D-SERINE

D-PHENYLALANINE

DL-2-FLUOROPHENYLALANINE

6-HYDROXYDOPA

DL-3 -FLUOROPHENYLALANINE

N-ACETYLGLYCINE

N-TRITYLGLYCINE

TRICINE

GUANIDOACETIC ACID

SARCOSINE

IMINODIACETIC ACID

ETHYLENEDIAMINE-N,N' -DIACETIC ACID

D-ALLO-THREONINE

N-CBZ-S-BENZYL-L-CYSTEINE

N-ACETYL-DL-PENICILLAMINE

DL-PENICI LAMINE

DL-CYSTEINE

DL-HOMOCYSTEINE

L-AZETIDINE-2 -CARBOXYLIC ACID

L-CARNOSINE

DL-HISTIDINE

L-THIAZOLIDINE-4-CARBOXYLIC ACID

3 , 4-DEHYDRO-DL-PROLINE

3 -AMINOPYRAZOLE-4-CARBOXYLIC ACID DL-PROLINE

CIS-4-HYDROXY-D-PRO INE

DL-ALPHA- (2 -THIENYL) GLYCINE

BETA-2 -THIENYL-DL-ALANINE

L-ABRINE

D-TRYPTOPHAN

5-BROMO-DL-TRYPTOPHAN

5-FLUORO-DL-TRYPTOPHAN

DL-5-METHOXYTRYPTOPHAN

DL-5-HYDROXYTRYPTOPHAN

5-METHYL-DL-TRYPTOPHAN

6-FLUORO-DL-TRYPTOPHAN

1-METHYL-DL-TRYPTOPHAN

L-PIPECOLINIC ACID

NIPECOTIC ACID

ISONIPECOTIC ACID

NITRO-L-ARGININE

3 -NITRO-L-TYROSINE

4-NITROHIPPURIC ACID

DL-HOMOCYSTEIC ACID

P-AMINOOXANILIC ACID

4-AMINOHIPPURIC ACID

ALBIZZIIN

DL-CITRULLINE

S-CARBAMYL-L-CYSTEINE

N-ALPHA-CBZ-L-ASPARAGINE

D-ASPARAGINE

N-CBZ-L-GLUTAMINE

L-GLUTAMINE

2 , 2-DIPHENYLGLYCINE

2-AMINOISOBUTYRIC ACID

D- ( - ) -ALPHA-PHENYLGLYCINE

DL-3-AMINO-3-PHENYLPROPIONIC ACID

D-ALANINE

DL-3-AMINOBUTYRIC ACID

DL-2-AMINOBUTYRIC ACID

D-NORVALINE

D-NORLEUCINE

DL-2-AMINO-N-CAPRYLIC ACID

DL-2-AMINO-N-CAPRYLIC ACID

GLYCINE

DL-ISOSERINE

DL-4-AMINO-3-HYDROXYBUTYRIC ACID

DL-3-AMINOISOBUTYRIC ACID

BETA-ALANINE

4-AMINOBUTYRIC ACID

5-AMINOVALERIC ACID

N-ALPHA-ACETYL-L-LYSINE

Z-L-VALINE

N- (4-NITROBENZOYL) -BETA-ALANINE

N- (4-AMINOBENZOYL) -BETA-ALANINE 1-AMINOCYCLOPROPANE-l-CARBOXYLIC ACID (R) - (-) -N- (3 , 5-DINITROBENZOYL) -ALPHA-PHENYLGLYCINE N- (3 , 5-DINITROBENZOYL) -DL-LEUCINE (+/-)-3-AMINOADIPIC ACID ALPHA-METHYL-DL-PHENYLALANINE DL-2-AMINO-5-PHOSPHONOVALERIC ACID DL-3 -HYDROXYNORVALINE HYDRATE (+/-)-INDOLINE-2-CARBOXYLIC ACID BETA- (2-THIAZOLYL) -DL-ALANINE L-GAMMA-CARBOXYGLUTAMIC ACID

L-2- (BENZYLOXYCARBONYLAMINO) -4-SULFAMOYLBUTYRIC ACID L-2-AMINO-4-SULFAMOYLBUTYRIC ACID

1-AMINO-1-CYCLOPROPANECARBOXYLIC ACID HYDROCHLORIDE 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

2-AMINO-3-PHENYLBUTANOIC ACID HYDROCHLORIDE (+/-)-l,2,3, 4-TETRAHYDRO-3 -ISOQUINOLINECARBOXYLIC

ACID HYDROCHLORIDE DL-PIPECOLINIC ACID HYDROCHLORIDE 5-AMINOLEVULINIC ACID HYDROCHLORIDE GLYCINE HYDROCHLORIDE 5-HYDROXY-DL-LYSINE HYDROCHLORIDE DL-2,3-DIAMINOPROPIONIC ACID HYDROBROMIDE DL-2,3-DIAMINOPROPIONIC ACID MONOHYDROCHLORIDE DL-2,4-DIAMINOBUTYRIC ACID DIHYDROCHLORIDE D-ORNITHINE HYDROCHLORIDE D-ORNITHINE HYDROCHLORIDE 5-AMINOVALERIC ACID HYDROCHLORIDE D-LYSINE MONOHYDROCHLORIDE DL-2-AMINO-4-PHOSPHONO BUTYRIC ACID DL-2-AMINO-4-PHOSPHONO BUTYRIC ACID N-PHENYLGLYCINE

DL-2-AMINO-3-PHOSPHONOPROPIONIC ACID N-CBZ-L-PHENYLALANINE N-(CARBOXYMETHYL) ANTHRANILIC ACID N- ( P-TOLUOYL) -GLYCINE L-ASPARTIC ACID MONOPOTASSIUM SALT L-ASPARTIC ACID MONOPOTASSIUM SALT N-CARBOBENZYLOXY-L-LEUCINE N-CARBOBENZYLOXY-L-ISOLEUCINE

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 4-CHLORO-L-PHENYLALANINE P-IODO-D-PHENYLALANINE O-PHOSPHO-DL-TYROSINE D-TYROSINE DL-TYROSINE 3 -FLUORO-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

DL-2-AMINO-4-PENTENOIC ACID 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 L-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

CIS-4-HYDROXY-L-PROLINE L-HYDROXYPROLINE BETA- (2-THIENYL) -L-ALANINE DL-TRYPTOPHAN L-TRYPTOPHAN L-5-HYDROXYTRYPTOPHAN 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- YSINE

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 MONOHYDROCHLORIDE L-2,4-DIAMINOBUTYRIC ACID DIHYDROCHLORIDE L-ORNITHINE MONOHYDROCHLORIDE DL-LYSINE MONOHYDROCHLORIDE L-LYSINE MONOHYDROCHLORIDE N- (TERT-BUTOXYCARBONYL) -L-ASPARTIC ACID 4-BENZYL ESTER N-ALPHA- (TERT-BUTOXYCARBONYL) -L-GLUTAMINE N-ALPHA- (TERT-BUTOXYCARBONYL) -L-HISTIDINE BOC-LEU H20 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 3-CHLORO-L-ALANINE HYDROCHLORIDE (S) -(-) -INDOLINE-2 -CARBOXYLIC ACID

N- ( PHOSPHONOMETHYL) GLYCINE, MONOISOPROPYLAMINE SALT MIMOSINE D-HOMOSERINE ( + ) -OCTOPINE

N-CARBOBENZYLOXY-L-GLUTAMIC ACID 1-METHYL ESTER DL-CYSTINE

3- (CARBOXYMETHYLAMINOMETHYL) -4-HYDROXYBENZOIC ACID CIS-3 -HYDROXY-DL-PROLINE (S)-(-)-l,2,3, 4-TETRAHYDRO-3 -ISOQUINOLINECARBOXYLIC

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

SESQUIHYDRATE DL-2-METHYLGLUTAMIC ACID HEMIHYDRATE N-METHYL-D-ASPARTIC ACID MONOHYDRATE DL-3-PHENYLSERINE HYDRATE 3-METHYL-L-HISTIDINE HYDRATE

N-(4-NITROBENZOYL) -L-GLUTAMIC ACID HEMIHYDRATE 4-NITRO-DL-PHENYLALANINE HYDRATE L-CYSTEIC ACID MONOHYDRATE 4-AMINO-DL-PHENYLALANINE HYDRATE D-ASPARAGINE MONOHYDRATE D-LYSINE HYDRATE

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

MONOHYDRATE 2-METHYLORNITHINE HYDROCHLORIDE MONOHYDRATE 3-AMINO-L-TYROSINE DIHYDROCHLORIDE MONOHYDRATE 4-AMINO-L-PHENYLALANINE HYDROCHLORIDE HEMIHYDRATE L-GLUTAMIC ACID, MONOSODIUM SALT, MONOHYDRATE 3 , 5-DIIODO-L-TYROSINE DIHYDRATE

N- ( 9-FLUORENYLMETHOXYCARBONYL) -L-ALANINE MONOHYDRATE 3 , 5-DINITRO-L-TYROSINE MONOHYDRATE 4-NITRO-L-PHENYLALANINE MONOHYDRATE DL-GLUTAMIC ACID MONOHYDRATE L-CYSTEINESULFINIC ACID MONOHYDRATE KAINIC ACID MONOHYDRATE 4-AMINO-L-PHENYLALANINE HYDRATE IBOTENIC ACID MONOHYDRATE

L-HISTIDINE MONOHYDROCHLORIDE MONOHYDRATE DL-HISTIDINE MONOHYDROCHLORIDE MONOHYDRATE DL-CYSTEINE HYDROCHLORIDE MONOHYDRATE DL-ARGININE HYDROCHLORIDE MONOHYDRATE L-LYSINE MONOHYDRATE L-ASPARAGINE MONOHYDRATE DL-ASPARAGINE MONOHYDRATE

(S)-(-) -ALPHA-AMINOCYCLOHEXANEPROPIONIC ACID HYDRATE N-BENZYLGLYCINE HYDROCHLORIDE 2-AMINO-2-NORBORNANECARBOXYLIC ACID CIS-4-AMINO-l-CYCLOHEXANECARBOXYLIC ACID

(+/- ) -TRANS-3-AZABICYCLO (3.1.0) HEXANE-2-CARBOXYLIC ACID 3-AZETIDINECARBOXYLIC ACID N-CARBOBENZYLOXY-L-TYROSINE 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 = 0, an aminothioether acid, a compound of the formula

P N Q—CH X

II

Rn is reacted with about 1 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, R_n, 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

3 -MERCAPTOPROPIONIC ACID

DL-HOMOCYSTEINE

2-MERCAPTONICOTINIC ACID

D-CYSTEINE HYDROCHLORIDE

3-MERCAPTOBENZOIC ACID

4-MERCAPTOBENZOIC ACID

DL-2-MERCAPTOMETHYL-3 -

GUANIDINOETHYLTHIOPROPANOIC ACID

2-THIOURACIL-5-CARBOXYLIC ACID

L-THIOHISTIDINE

4-MERCAPTOBUTYRIC ACID

CYS-GLY

DL-THIORPHAN 4-MERCAPT0HYDR0CINNAMIC 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

6-AZA-2-THIOURACIL-5-CARBOXYLIC ACID

2-THIOACETIC ACID-5-MERCAPTO-1 , 3 , 4-

THIADIAZOLE

N-ISOBUTYRYL-L-CYSTEINE

N-ISOBUTYRYL-D-CYSTEINE

CAPTOPRIL

5-MERCAPTO-TETRAZOLE-l-PROPIONIC ACID

SALOR S98, 217-2

2,6-CSBA (2-Cl-6-mercaptobenzoic acid)

6-MERCAPTONICOTINIC ACID

MAYBRIDGE RJC 01025, and the like.

In the preparation of aminothioether acid compounds the mercaptide salt is reacted with a compound of the

Formula II. The substituent X is a good leaving group subject to nucleophilic displacement by the mercaptide salt. Exemplary of suitable X groups include, but are not intended to be limited to, mesylate, tosylate, halo, and the like. Compounds of Formula II are typically derived from an amino alcohol or protected-amino alcohol of the formula p _N Q _CH0H

wherein P, R_n, Q and R_s 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:

TRANS-2 -AMINOCYCLOHEXANOL HYDROCHLORIDE 3 -AMINOMETHYL-3 , 5 , 5-TRIMETHYLCYCLOHEXANOL (1R,2R) -(-)-PSEUDOEPHEDRINE -ADRENALINE DL-HOMOSERINE Z-L-SERINE 2 -ANILINOETHANOL N-ACETYLETHANOLAMINE 2- (METHYLAMINO) ETHANOL N-BENZYLETHANOLAMINE 2- (ETHYLAMINO) ETHANOL DIETHANOLAMINE 2 - ( PROPYLAMINO) ETHANOL D-SERINE

(lS,2S)-(+) -2-AMINO-l-PHENYL-l,3-PROPANEDIOL D-ALLO-THREONINE DIISOPROPANOLAMINE 2-AMINO-2-METHYL-1 , 3-PROPANEDIOL TRIS (HYDROXYMETHYL) AMINOMETHANE N-METHYL-D-GLUCAMINE DL-2 -AMINO-3 -METHYL-1-BUTANOL L-ISOLEUCINOL L-PHENYLALANINOL DL-4-CHLOROPHENYLALANINOL L-METHIONINOL CIS-4-HYDROXY-D-PROLINE L-PROLINOL 3-PYRROLIDINOL 3-PYRROLIDINOL 2-PIPERIDINEMETHANOL 2 -PIPERIDINEETHANOL 3 -PIPERIDINEMETHANOL 4-HYDROXYPIPERIDINE 2 -AMINO-2-METHYL-1-PROPANOL D- ( - ) -ALPHA-PHENYLGLYCINOL (-) -NOREPHEDRINE DL-2-AMINO-l-PROPANOL (+/-) -2-AMINO-l-BUTANOL 2 - MINO-1-PHENYLETHANOL DL-ISOSERINE 1-AMINO-2 -PROPANOL 3 -AMINO-1 , 2 -PROPANEDIOL DL-4-AMINO-3-HYDROXYBUTYRIC ACID 1 , 3 -DIAMINO-2 -PROPANOL 2 - (2 -AMINOETHYLAMINO) ETHANOL ETHANOLAMINE 3 -AMINO-1-PROPANOL 4-AMINO-1-BUTANOL 5-AMINO-1-PENTANOL 6-AMINO-1-HEXANOL DL-2 -AMINO-1-PENTANOL DL-2-AMINO-l-HEXANOL 1-AMINO-1-CYCLOPENTANEMETHANOL

(1S,2S) -(+) -2-AMINO-3-METHOXY-1-PHENYL-1-PROPANOL DL-PROPRANOLOL HYDROCHLORIDE TRANS-4-AMINOCYCLOHEXANOL HYDROCHLORIDE TRIS (HYDROXYMETHYL) AMINOMETHANE HYDROCHLORIDE DL-SERINE METHYL ESTER HYDROCHLORIDE L-SERINE ETHYL ESTER HYDROCHLORIDE L-PHENYLEPHRINE HYDROCHLORIDE 3 -HYDROXYPIPERIDINE HYDROCHLORIDE DL-OCTOPAMINE HYDROCHLORIDE DL-NORMETANEPHRINE HYDROCHLORIDE ETHANOLAMINE HYDROCHLORIDE 3 -HYDROXYPIPERIDINE N-CYCLOHEXYLETHANOLAMINE L-NORADRENALINE L-ADRENALINE BITARTRATE 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-PROPANOL HYDROCHLORIDE 2-AMINO-1,3 -PROPANEDIOL OXALATE

METHYL 3-AMINO-3-DEOXY-BETA-D-GLUCOPYRANOSIDE 2-AMINO-1, 3-PROPANEDIOL DL-SERINE HYDROXAMATE L-SERINE BETA-NAPHTHYLAMIDE N-CBZ-D-GLUCOSAMINE

N- (TERT-BUTOXYCARBONYL) ETHANOLAMINE DL-THREONINE HYDROXAMATE SER-BETA-ALA N-T-BOC-L-HOMOSERINE

METHYL 3 -AMINO-3 -DEOXY-ALPHA-D-MANNOPYRANOSIDE HYDROCHLORIDE

L-THREONINAMIDE HYDROCHLORIDE 3-AMINO-2 , 2-DIMETHYL-1-PROPANOL 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 CIS-4-HYDROXY-L-PROLINE L-HYDROXYPROLINE D-PROLINOL

L- (+) -ALPHA-PHENYLGLYCINOL (1S,2R) - (+) -PHENYL-PROPANOLAMINE (S) -(+) -2 -AMINO-1-PROPANOL D-ALANINOL

(S)-(+) -2-AMINO-l-BUTANOL (R) - (-) -2-AMINO-l-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-l,2-DIPHENYLETHANOL (1R,2S) -(-) -2-AMINO-l,2-DIPHENYLETHANOL S-BENZYL-L-CYSTEINOL BOC-L-PHENYLALANINOL L-HISTIDINOL DIHYDROCHLORIDE

D-(-)-THREO-2-AMINO-l-(4-NITROPHENYL) -1,3- PROPANEDIOL

ETHAMBUTOL DIHYDROCHLORIDE (+/-) -ARTERENOL BITARTRATE SALT S-ALPHA-HYDROXYMETHYL TYROSINE ( 1S , 2S ) - ( + ) -THIOMICAMINE

CIS-2-AMINOMETHYL-l-CYCLOHEXANOL HYDROCHLORIDE CIS-2 -HYDROXYMETHYL-1-CYCLOHEXYLAMINE HYDROCHLORIDE TRANS-2 -AMINOMETHYL-1-CYCLOHEXANOL TRANS-2-HYDROXYMETHYL-1-CYCLOHEXYLAMINE

HYDROCHLORIDE

DL-3-PHENYLSERINE HYDRATE

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 3-PHENYL-DL-SERINE 3-PHENYL-DL-SERINE

(+/-) -NOREPINEPHRINE L-BITARTRATE HYDRATE

(1S,2R) -(-) -CIS-l-AMINO-2-INDANOL 4-AMINO-2-BUTANOL

2- [2- (AMINOPHENYLTHIO] BENZYL 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

I III n

wherein P, R_n and Q are as defined above. Available amino acid compounds can be protected with an amino-protecting group and converted in good yields to the corresponding protected-amino alcohols of the formula

P N Q CHOH

Rn R

For example, when R_s 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 = R_n = hydrogen and Q =

wherein Ri 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 steps proceed smoothly and can be carried out on multigram scale.

3b

3a P=Fmoc

P=Alloc P=Alloc or Boc or Boc

Scheme II a) Allyl chloroformate or di-t-butyl dicarbonate, Na2Cθ3,

1:1 H2θ:dioxane, 4°C, 90-95%, P=Alloc or Boc, respectively; b) isobutyryl chloroformate, N-methylmorpholine (NMM) in ethylene glycol dimethyl ehter (DME) , -15°C, then aq NaBH4, -15°C, 65-90%; c) CH3SO2CI, TEA in THF, 4°C, 90-95%; d) mercapto acid (leq) , NaOMe (2eq) in DMF, r.t., 65-95%; e) P= Alloc→Fmoc, PPh3 (0.25eq), Pd(PPh3)4 (0.03eq), HOAc (2eq) in THF, then Fmoc-N-hydroxysuccinimide ester,

Na2C03 in 1:1 H2O: dioxane, 4°C to r.t., 50%; P=

Boc→Fmoc, 4N HCl in dioxane or EtOAc, 1.5h, then Na2C03 and F oc-OSu in 1:1 H2O: dioxane, 90-95%.

The aminothioether compounds can be converted to their corresponding sulfoxides (m=l) 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, periodate, 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 92-100% yield] .

2) . D. M. Hedstrand, S. R. Byrn, A. T. McKenzie and P. L. Fuchs J. Org. 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 corresponding sulfone:

4). B. M. Trost and R. Braslau J. Org. 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 1

_L COOH

2 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 acid with the terminal amino group on the solid support bound product is carried out under essentially the same reaction 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 carbodiimides to form the intermediate "active" HOBT ester 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" 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

HN T N CO— CH₂-C₆H4-0-CH₂- C₆H - Polymer

Ky Ky" the product macrocycle library compounds of the invention are those of Formula I wherein B is a group of the formula

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 agents include, but are not intended to be limited to, organic halides, acyl halides, sulfonic acid esters, organohaloformates, organosulfonyl halides, organic isocyanates, organic isothiocyanates, aldehydes, ketones, and the like. Examples of such electrophilic agents include, but are not intended to be limited to:

3 , 5-bis ( trifluoromethyl) benzoyl chloride benzoyl chloride

2-bromobenzoyl chloride

2-fluorobenzoyl chloride pentafluorobenzoyl chloride

2 , 4-difluorobenzoyl 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 chloride

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 pal itoyl 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-bromohexanoy1 chloride pimeloyl chloride heptanoyl chloride suberoyl chloride octanoyl chloride

10-undecenoyl chloride

2 -chloro-2 , 2 -diphenylacetyl chloride dichloroacetyl chloride alpha-chlorophenylacetyl chloride

2-chloropropionyl chloride

2-iodobenzoyl chloride

4-iodobenzoyl chloride cyclopropanecarbonyl chloride trans-2 -phenyl-1-cyclopropanecarbonyl chloride cyclobutanecarbonyl chloride 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-dimethoxyphenylacetyl chloride

3-methoxyphenylacetyl chloride iminodibenzyl-5-carbonyl chloride

2 , 4, 6-trimethylbenzoyl chloride tetrafluorosuccinyl chloride perfluorooctanoyl chloride diphenylacetyl chloride alpha-methyl valeroyl chloride methyl alonyl 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-l-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-l-naphthoyl chloride pyrrolidine carbonyl chloride m- (chlorosulfonyl) benzoyl chloride

2-n-propyl-n-valeroyl chloride 2-chloro-4-nitrobenzoyl chloride

2 -phenoxybutyry1 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-trifluoropropionylchloride

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 -carbonyl chloride carbazole-n-carbonyl chloride

2-nitrophenoxyacetyl chloride

2-bromo-2-methylpropionyl chloride

2-fluoro-3- (trifluoromethyl) benzoyl chloride 2-fluoro-4- (trifluoromethyl) benzoyl chloride

2-fluoro-5- (trifluoromethyl) 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 (trifluoromethyl) benzoyl chloride 3- (trifluoromethoxy) benzoyl chloride m- (fluorosulfonyl ) benzoyl chloride trans-1, 2-cyclobutanedicarboxylic acid chloride 3-cyclohexylpropionyl chloride

4-ethyl-2 , 3-dioxo-l-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 l-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-di ethylsuccinyl chloride alpha-bromoisovaleryl chloride benzoyl chloride oleoyl chloride methyl suberyl chloride gamma-linolenoyl chloride

(-) -camphanic acid chloride 4, 4 ' -stilbenedicarbonyl chloride chlorinated benzoyl chloride

(lr) -(+) -camphanic chloride

2- (4-nitrophenoxy) tetradecanoyl chloride

7- [ (chlorocarbonyl)me hoxy] -4-methylcoumarin n,n-bis (2 -chloroethyl) carbamoyl chloride

(s) - (-) -2-acetoxypropionyl chloride linoleoyl chloride

3 -chlorotetrafluoropropionyl 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-naphthalenesulfonyl) -1-phenylalanyl chloride n- (4-nitrophenylsulfonyl) -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 l-bromo-2-butanone l-bromo-2 , 2-dimethoxypropane l-bromo-2 , 2-dimethylpropane bromoacetaldehyde dimethyl acetal bromoacetaldehyde diethyl acetal l-bromo-2-methylpropane l-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-bromomethy1-6,7-dimethoxycoumarin 2, 4-difluorobenzyl bromide methyl 2- (bromomethyl) acrylate

3-bromopropionaldehyde dimethyl acetal.

(r) - (-) -3-bromo-2-methyl-l-propanol, and the like.

Sulfonic Acid Esters ethyl trifluoromethanesulfonate 2,2, 2-trifluoroethyl p-toluenesulfonate 2-chloroethyl-p-toluenesulfonate 1 , 3 -propane sultone 5 ' -tosyladenosine

1,4-butane sultone cyanomethyl benzenesulfonate hexadecyl methanesulfonate ethyl methanesulfonate

2-chloroethyl methanesulfonate ethyl p-toluenesulfonate trans-2 -hydroxycyclohexyl p-toluenesulfonate

(2r) -(-) -glycidyl tosylate (s) - (+) -2-methylbutyl methanesulfonate

(s) - (+ ) -2-methylbutyl p-toluenesulfonate

(s) - (+) -1-phenyl-1, 2-ethanediol 2 -tosylate

(2r) - (-) -glycidyl 3-nitrobenzenesulfonate propargyl benzenesulfonate 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-o- isopropylidene-alpha-d-glucofu

(r) - (-) -l-benzyloxy-3- (p-tosyloxy) -2-propanol (s) - (+) -l-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 chloroformate phenyl chloroformate 4-chlorophenyl chloroformate methyl chloroformate benzyl chloroformate vinyl chloroformate isobutyl chloroformate 2-ethylhexyl chloroformate 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-l, 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 l-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-triisopropylbenzenesulfonyl chloride

2-mesitylenesulfonyl chloride 3-nitrobenzenesulfonyl chloride p-bromobenzenesulfonyl chloride

4-fluorobenzenesulfonyl chloride

4-chlorobenzenesulfonyl chloride

4-chloro-3-nitrobenzenesulfonyl chloride 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- (chlorosulfonyl) benzoate

2-nitro-4- (trifluoromethyl) benzenesulfonyl chloride 3- (trifluoromethyl) benzenesulfonyl chloride

1-octanesulfonyl chloride

4- (trifluoromethoxy)benzenesulphonyl chloride (lr) - (-) -10-camphorsulfonyl chloride d- (+ ) -10-camphorsulfonyl chloride (+/-) -10-camphorsulfonyl chloride

2-nitro-alpha-toluenesulfonyl 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 isocyanate

2 , 4-dichlorophenyl isocyanate 2 , 5-dichlorophenyl isocyanate

2 , 6-dichlorophenyl isocyanate 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 -methoxycarbonylphenyl isocyanate -ethoxycarbonylphenyl isocyanate -isopropylphenyl isocyanate , 3 -dimethylphenyl isocyanate -methoxy-2-methylphenyl isocyanate , 4-dimethylphenyl isocyanate , 5-dimethylphenyl isocyanate -ethyl-6-methylphenyl isocyanate -cyanophenyl isocyanate -chloro-2 , 4-dimethoxyphenyl isocyanate -chloro-4-methylphenyl isocyanate , 5-dichlorophenyl isocyanate -chloro-2 -methoxyphenyl isocyanate ,4, 5-trimethoxyphenyl isocyanate , 5-dimethoxyphenyl isocyanate - (methylthio) phenyl isocyanate -ethoxycarbonylphenyl isocyanate -acetylphenyl isocyanate , 4-dimethylphenyl isocyanate 3 , 5-dimethylphenyl isocyanate -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 isocyanate 4-ethylphenyl isocyanate 4-n-butylphenyl isocyanate 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 -bro opropyl isocyanate 2 , 4, 6-trimethylphenyl isocyanate 2 -isopropyl-6-methyl henyl isocyanate 2 , 6-diethylphenyl isocyanate

5-chloro-2-methylphenyl isocyanate 4-chloro-2-methylphenyl isocyanate 4- (trifluoromethoxy) phenyl isocyanate 4-trifluoromethylthiophenylisocyanate 2, 4-dibromophenyl isocyanate

2, 6-dibromo-4-ethylphenyl 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-isocyanatopropionate 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) enzene n-amyl isocyanate n-heptyl isocyanate

2-chloroethyl isocyanate, [ethyl-1, 2-14c] 1,1,3, 3-tetramethylbutyl isocyanate 3 , 5-dinitrophenyl isocyanate, and the like.

Aldehydes :

Ethyl 2-formyl-l-cyclopropanecarboxylate

Cyclohexanecarboxaldehyde

1,2,3, 6-Tetrahydrobenzaldehyde

Diphenylacetaldehyde

2-Phenylpropionaldehyde 2 , 3-Dimethylvaleraldehyde

Isobutyraldehyde

2 , 6-Dimethyl-5-hepten-l-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-l-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-Tri ethylhexanal

Succinic semialdehyde

(+/-) -3-Phenylbutyraldehyde

2,6, 6-Trimethyl-l-cyclohexene-l-acetaldehyde Cyclopropanecarboxaldehyde

3 -Cyclohexylpropionaldehyde

Hydroxycitronellal

Cis-4-heptenal

Cis-6-nonen-l-al Tetrahydrocitral

Cis-7-decen-l-al

Cis-8-undecen-l-al

3,5, 6-Trimethyl-3-cyclohexene-l-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 for yl succinate

4-Pregnene-20-beta-carboxaldehyde-3-one

Cis-7-tetradecenal

Cyclopentylmethanal 3, 4-Dimethyl-3 -eyelohexenylmethanal

2,4, 6-Trimethyl-3-cyclohexen-l-carboxaldehyde

Adipic semialdehyde methyl ester

Cis-14-methyl-8-hexadecenal

Cis-3-hexen-l-al Trans-4-decen-l-al

2 , 2-Dichlorooctadecanal 2 , 2-Dichlorotetradecanal

2 , 2 -Dichlorooctanal

2 , 2-Dichlorohexanal

(r) - (+) -Citronellal 8-Methyl-7-nonenal

2- (p-Tolyl) propionaldehyde

Aldehyde C-ll MOA (2-methyldecanal)

Alpha-methylhydrocinnamaldehyde

(s) - (-) -Citronellal 4-Hydroxybutanal

4-Oxobutyric acid methyl ester

3,3,4,4,5,5, 5-Heptafluoropentanal

3-Methylbutanal-l-13c

6-Methyl-3 -cyclohexene-1-carboxaldehyde 4- (4-Methyl-2-pentenyl) -3-cyclohexene-l- carboxaldehyde

3-Pentyn-l-al

3-Pyridylacetaldehyde n-oxide

2 , 3-Dihydro-5-methoxy-3-phenyl-2- indolecarboxaldehyde

2, 4-Diphenyl-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 -Acetoxypropionaldehyde 2-Methyl-4-phenylpentanal (lr, 2s, 3r, 4s) - (+) -2-Benzyloxy-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 l-Formyl-6- (dimethylamino) fulvene

1 , 4-Dioxaspiro (4.5) decane-7-acetaldehyde Alpha-citronellal

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, [l-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, lO-Dihydro-9, 10-ethanoanthracene-ll-carboxaldehyde Methyl hexyl acetaldehyde

2 , 3-Dihydro-2-oxo-lH-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:

Ketones :

1-BROMO-2 -BUTANONE 2-FLUOROPHENYLACETONE 4-FLUOROPHENYLACETONE 3 -TRIFLUOROMETHYLPHENYLACETONE 1,1, l-TRIFLUORO-2 , 4-PENTANEDIONE FLUOROACETONE 4 ' -CHLOROACETOACETANILIDE 3 -CHLORO-2 -BUTANONE CHLOROACETONE 1 , 3 -DICHLOROACETONE METHYL 4-CHLOROACETOACETATE ETHYL 4-CHLOROACETOACETATE 1-CHLORO-3 -PENTANONE 5-CHLORO-2 -PENTANONE CYCLOTRIDECANONE CYCLOPENTADECANONE DICYCLOPROPYL KETONE CYCLOPROPYL METHYL KETONE (-) -THUJONE CYCLOBUTANONE 2-CYCLOPENTEN-1-ONE 3 -METHYL-2 -CYCLOPENTEN-1-ONE 4-CYCLOPENTENE-1, 3 -DIONE 1 , 3 -CYCLOPENTANEDIONE 2 -METHYL-1 , 3 -CYCLOPENTANEDIONE CYCLOPENTANONE 2 -CHLOROCYCLOPENTANONE

CYCLOPENTANONE-2-CARBOXYLIC ACID METHYL ESTER ETHYL 2 -OXOCYCLOPENTANECARBOXYLATE 2-ACETYLCYCLOPENTANONE 2 -METHYLCYCLOPENTANONE 2 , 4-DIMETHYLCYCLOPENTANONE 3 -METHYLCYCLOPENTANONE BETA-IONONE

BETA-IONONE

ALPHA-IONONE

2-CYCLOHEXEN-l-ONE

3-ETHOXY-2-CYCLOHEXEN-1-ONE

3 -METHYL-2 -CYCLOHEXEN-1-ONE

4-CARBETHOXY-3-METHYL-2-CYCLOHEXEN-1-ONE

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

ISOPHORONE

1 , 3-CYCLOHEXANEDIONE

2 -ACETYL-1 , 3 -CYCLOHEXANEDIONE

2 -METHYL-1 , 3 -CYCLOHEXANEDIONE

DIMEDONE

1 , 4-CYCLOHEXANEDIONE

DIMETHYL 1 , 4-CYCLOHEXANEDIONE-2 , 5-DICARBOXYLATE

DIETHYL 1 , 4-CYCLOHEXANEDIONE-2 , 5-DICARBOXYLATE

CYCLOHEXANONE

2 -CHLOROCYCLOHEXANONE

2 -NITROCYCLOHEXANONE

2 -PHENYLCYCLOHEXANONE

2 - ( 3 -METHOXYPHENYL) CYCLOHEXANONE

ETHYL 2-CYCLOHEXANONECARBOXYLATE

ETHYL 4-METHYL-2-CYCLOHEXANONE-1-CARBOXYLATE

2 -ACETYLCYCLOHEXANONE

L-MENTHONE

2 -METHYLCYCLOHEXANONE

( + ) -DIHYDROCARVONE

2 , 6-DIMETHYLCYCLOHEXANONE

ETHYL-2 -CYCLOHEXANONE ACETATE

3 -METHYLCYCLOHEXANONE

3,3,5, 5-TETRAMETHYLCYCLOHEXANONE

4-PHENYLCYCLOHEXANONE

4-TERT-BUTYLCYCLOHEXANONE

4-METHYLCYCLOHEXANONE

4-ETHYLCYCLOHEXANONE

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

NAPHTHALENEDIONE

2-ACETYL-1-TETRALONE

1-DECALONE

BETA-TETRALONE

1-METHYL-2 -TETRALONE

6-METHOXY-2-TETRALONE

7-METHOXY-2 -TETRALONE

2 -DECALONE

CYCLOOCTANONE

CYCLONONANONE

ALPHA-ACETYL-PHENYLACETONITRILE

ANTI-PYRUVIC ALDEHYDE 1-OXIME

4- (4-HYDROXYPHENYL) -2 -BUTANONE

1 , 3-ACETONEDICARBOXYLIC ACID LEVULINIC ACID

4,6-DIOXOHEPTANOIC ACID

4-KETOPIMELIC ACID

3 -ACETYL-1-PROPANOL

4-ANDROSTENE-3 , 17-DIONE

CYCLODECANONE

CYCLOUNDECANONE

CYCLODODECANONE

CIS-BICYCLO (3.3.0) OCTANE-3 , 7-DIONE

CIS-l,5-DIMETHYLBICYCLO(3.3.0 ) OCTANE-3 , 7-DIONE

2 -INDANONE

2 -CYCLOHEPTEN-1-ONE

CYCLOHEPTANONE

4-ACETYLBUTYRIC ACID

4-HYDROXY-4-METHYL-2-PENTANONE

3 -HYDROXY-2 -BUTANONE

HYDROXYACETONE

4-HYDROXY-3 -METHYL-2 -BUTANONE l-BENZYL-3-PYRROLIDINONE

2-ACETYLBUTYROLACTONE

TETRAHYDROTHIOPHEN-3 -ONE

TROPINONE

6-HYDROXYTROPINONE

4-OXO-TEMPO

N-CARBETHOXY-4-PIPERIDONE

N-BENZOYL-4-PIPERIDONE l-ACETYL-4-PIPERIDONE l-METHYL-4-PIPERIDONE l-BENZYL-4-PIPERIDONE

1- (BETA-PHENYLETHYL) -4-PIPERIDONE

TETRAHYDRO-4H-PYRAN-4-ONE

1,4-CYCLOHEXANEDIONE MONO-2,2-

DIMETHYLTRIMETHYLENE

KETAL TETRAHYDROTHIOPYRAN-4-ONE METHYLGLYOXAL DIMETHYL ACETYLSUCCINATE DIMETHYL 1 , 3 -ACETONEDICARBOXYLATE DIMETHYL 3-OXOADIPATE 4- (4-ACETOXYPHENYL) -2 -BUTANONE METHYL 2 -CHLOROACETOACETATE METHYLGLYOXAL DIMETHYL ACETAL 1 , 1-DIPHENYLACETONE 1, 1-DIPHENYLACETONE METHYL 4-ACETYL-5-OXOHEXANOATE 3 -METHYLTHIO-2 -BUTANONE 3-METHYLTHIO-2-BUTANONE 3 -METHYL-2 , 4-PENTANEDIONE 3 -ETHYL-2 , 4-PENTANEDIONE ACETONE (DIMETHYLAMINO) ACETONE

PHENOXYACETONE

METHOXYACETONE

DIMETHYL ( 2 -OXOPROPYL) PHOSPHONATE

2 -METHOXYPHENYLACETONE

3-METHOXYPHENYLACETONE

3 , 4-DIMETHOXYPHENYLACETONE

4-METHOXYPHENYLACETONE

4-METHOXYPHENYLACETONE

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

5-HEXEN-2-ONE

N-TERT-BUTYLACETOACETAMIDE

TERT-BUTYL ACETOACETATE

4 , 4-DIMETHYL-2 -PENTANONE

MESITYL OXIDE

PHORONE

6-METHYL-5-HEPTEN-2-ONE

GERANYLACETONE

GERANYLACETONE

DIISOPROPYL KETONE

3 -METHYL-2 -BUTANONE

METHYL ISOBUTYL KETONE

6-METHYL-2 , 4-HEPTANEDIONE

2 , 6-DIMETHYL-4-HEPTANONE

5-METHYL-2 -HEXANONE

2-TRIDECANONE

DIETHYLAMINOACETONE

1-DIETHYLAMINO-3 -BUTANONE

5-DIETHYLAMINO-2 -PENTANONE

ETHYL 2,4-DIOXOVALERATE

ETHYL 2 -CHLOROACETOACETATE

ETHYL 2-BENZYLACETOACETATE

DIETHYL ACETYLSUCCINATE DIETHYL 2 -ACETYLGLUTARATE

ETHYL 2 -METHYLACETOACETATE

ETHYL ISOBUTYRYLACETATE

ETHYL ACETOACETATE

DIETHYL 1 , 3 -ACETONEDICARBOXYLATE

ETHYL LEVULINATE

DIETHYL 4-OXOPIMELATE

ETHYL 4-ACETYL-5-OXOHEXANOATE

ETHYL 4-ACETYLBUTYRATE

DIETHYL 3-OXOPIMELATE

3-PENTEN-2-ONE

2-METHYL-3 -PENTANONE

1-PHENYL-2 -BUTANONE

ETHYL VINYL KETONE

ETHYL PROPIONYLACETATE

2,4-HEXANEDIONE

3 -PENTANONE

3 -METHYL-2 -PENTANONE

5-METHYL-3 -HEPTANONE

2 -METHYL-3 -HEXANONE

2 -PENTANONE

ETHYL BUTYRYLACETATE

3 -HEXANONE

3 -HEXANONE

4-HEPTANONE

BUTYL LEVULINATE

2-METHYL-3 -HEPTANONE

2 -HEXANONE

3 -HEPTANONE

5-NONANONE

2-HEPTANONE

DIMETHYL ( 2 -OXOHEPTYL) PHOSPHONATE

3 -OCTANONE

6-UNDECANONE

2-OCTANONE

3 -NONANONE

4-DECANONE

7-TRIDECANONE

2 -NONANONE

3 -DECANONE

8-PENTADECANONE

2 -DECANONE

3 -UNDECANONE

9-HEPTADECANONE

2 -UNDECANONE

10-NONADECANONE

2 -METHOXYCYCLOHEXANONE

2 -METHOXYETHYL ACETOACETATE

3 -CHLOROACETYLACETONE

1, 1-DICHLOROACETONE 6 , 7 -DIMETHOXY-2 -TETRALONE 4, 4 -DIMETHYL-2 -CYCLOHEXEN-1-ONE 2 -TERT-BUTYLCYCLOHEXANONE N- (ACETOACETYL) GLYCINE

3 , 5-DIACETYLTETRAHYDROPYRAN-2 , 4 , 6-TRIONE ALLYL ACETOACETATE

4- (TRIMETHYLSILYLOXY) -3 -PENTEN-2-ONE ETHYL DIACETOACETATE (R) - ( - ) -4 , 4A, 5 , 6 , 7 , 8-HEXAHYDRO-4A-METHYL-

2 ( 3H) NAPHTHALENONE 4, 4-DIMETHOXY-2 , 5-CYCLOHEXADIEN-l-ONE 3-OXOADIPIC ACID TETRONIC ACID

METHYL 4-METHOXYACETOACETATE DI-TERT-BUTYL 1 , 3 -ACETONEDICARBOXYLATE 1, 4-CYCLOHEXANEDIONE MONOETHYLENEKETAL DIMETHYL ( 3 -PHENOXYACETONYL ) PHOSPHONATE 4-ACETOXY-2 -BUTANONE 3-NONEN-2-ONE 1-HYDROXY-2 -BUTANONE 5 -METHYL-1 , 3 -CYCLOHEXANEDIONE 2 -METHYLTETRAHYDROFURAN-3 -ONE 1-PROPYL-4-PIPERIDONE 5ALPHA-ANDROST-16-EN-3 -ONE

2 -CHLORO-5, 5-DIMETHYL-1, 3 -CYCLOHEXANEDIONE 4-ACETYL-1-METHYL-1-CYCLOHEXENE 4-ACETYL-1-METHYL-1-CYCLOHEXENE 4-HEXEN-3-ONE

4- (TERT-BUTYLDIMETHYLSILYLOXY) -3 -PENTEN-2-ONE 5-KETOHEXANENITRILE

ETHYL 2 - ( TRIMETHYLSILYLMETHYL ) ACETOACETATE ETHYL 4-HYDROXY-6-METHYL-2 -OXO-3 -CYCLOHEXENE-

1-CARBOXYLATE METHYL 4-ALLYL-3 , 5-DIOXO-l-CYCLOHEXANECARBOXYLATE METHYL 2 -0X0-1 -CYCLOHEPTANECARBOXYLATE METHYL ETHYL KETONE METHYL 3 -OXOPENTANOATE 5-METHYL-3-HEXEN-2-ONE

4- (4-HYDROXY-3 -METHOXYPHENYL) -3 -BUTEN-2-0NE (S) - (+) -2 , 3 , 7, 7A-TETRAHYDRO-7A-METHYL-1H-INDENE- 1,5(6H) -DIONE

2 -HYDROXYMETHYL-6-METHOXY-1 , 4-BENZOQUINONE 8-CYCLOHEXADECEN-1-ONE 8-MERCAPTOMENTHONE

2,2, 6, 6-TETRAMETHYL-4-PIPERIDONE HYDROCHLORIDE METHYL 4-0X0-3 -PIPERIDINECARBOXYLATE HYDROCHLORIDE ETHYL 1-BENZYL-3-OXOPIPERIDINE-4-CARBOXYLATE

HYDROCHLORIDE l-ETHYL-3-PIPERIDONE HYDROCHLORIDE METHYL 1-BENZYL-4-OXO-3-PIPERIDINE-CARBOXYLATE

HYDROCHLORIDE 1-BENZYL-3-CARBETHOXY-4-PIPERIDONE HYDROCHLORIDE DELTA-AMINOLEVULINIC ACID, METHYL ESTER

HYDROCHLORIDE 5-AMINOLEVULINIC ACID HYDROCHLORIDE 1 , 4-DIAMINO-2-BUTANONE DIHYDROCHLORIDE DIACETONAMINE HYDROGEN OXALATE ACETOACETIC ACID LITHIUM SALT S-TERT-BUTYL ACETOTHIOACETATE ETHYL 4-OXOCYCLOHEXANECARBOXYLATE l-ETHYL-4-PIPERIDONE

N-HYDROXYSUCCINIMIDYL ACETOACETATE 3 -BROMO-2 -BUTANONE 1 , 3 -DIBROMOACETONE (4-CHLOROPHENYLTHIO) PROPAN-2-ONE 3 -AMINO-2 -CYCLOHEXEN-1-ONE 2 - ( 1-CYCLOHEXENYL) CYCLOHEXANONE 2 - ( BETA-CYANOETHYL) CYCLOHEXANONE 4-TERT-AMYLCYCLOHEXANONE 3 -ACETYLACRYLIC ACID TROPONE

METHYLSULFONYLACETONE BIS (4-METHOXYBENZYLIDENE) ACETONE BIS ( 4-METHOXYBENZYLIDENE) ACETONE 3 , 4-DIMETHOXYBENZYLIDENEACETONE 5-METHYL-5-HEXEN-2-ONE 2 , 6-DIMETHYL-3 , 5-HEPTANEDIONE 2 -DODECANONE 3 , 5-HEPTANEDIONE 2,4-OCTANEDIONE 2,4-NONANEDIONE 5-DODECANONE 4-DODECANONE 3 -DODECANONE (METHYLTHIO) ACETONE (+/-) -2-ETHOXYCYCLOHEXANONE 3-HEPTEN-2-ONE 3-OCTEN-2-ONE

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

2 ' , 5 ' -DICHLOROACETOACETANILIDE 4-CHLOROPHENYLSULFONYLACETONE 4-CHLOROBENZYLIDENEACETONE 2,4, 4-TRIMETHYLCYCLOPENTANONE CYCLOHEXYLACETONE 3,3, 5-TRIMETHYLCYCLOHEXANONE 4-PYRAZOLINO-2 -BUTANONE ACETOACETAMIDE 3-PHENYLAZOACETYLACETONE 3-N-BUTYL-2 , 4-PENTANEDIONE 3 -ACETYLOCTANONE-2 SALOR S17, 446-7 (BENZYLTHIO) ACETONE 6-PHENYLHEXA-3 , 5-DIEN-2-ONE 3-TRIDEC NONE 5-PENTADECANONE 7-HEPTADECANONE 4-HEPTADECANONE 3 -OCTADECANONE N, N-DIETHYLACETOACETAMIDE ETHYL 2 - ( PHENYLAZO) ACETOACETATE DIETHYL ACETOMALONATE ETHYL-2 -N BUTYLACETOACETATE (ETHYLTHIO) ACETONE 1-PHENYL-2 -PENTANONE 1-PHENYL-2 -HEXANONE N-AMYL ISOPROPYL KETONE 7-PENTADECANONE 4-TRIDECANONE 5-TETRADECANONE 6-PENTADECANONE 7-HEXADECANONE 1-OCTEN-3-ONE NOOTKATONE

CYCLOBUTYL METHYL KETONE l-ISOPROPYL-4-PIPERIDONE N, N-DIMETHYLACETOACETAMIDE FURFURALACETONE ACETOACET-M-XYLIDIDE ETHYL 2-ETHYLACETOACETATE DICYCLOHEXYL KETONE CYCLOHEXYL METHYL KETONE

1,2:5, 6-DI-0-ISOPROPYLIDENE-ALPHA-D-RIBO-3- HEXOFURANOSULOSE 4-HYDROXYPYRIDINE BENZYL ISOPROPYL KETONE 1,1,3 -TRICHLOROACETONE METHYL BUTYRYLACETATE CHLORODIFLUOROACETYLACETONE l-(2-FURYL) -1,3-BUTANEDIONE 4, 6-NONANEDIONE 2 -HEXADECANONE

N, N-DIMETHYL-2-CHLOROACETOACETAMIDE 3-ETHOXY-2-CYCLOPENTEN-1-ONE 2, 6, 6-TRIMETHYL-2-CYCLOHEXENE-1, 4-DIONE ETHYL 4-PIPERIDONE-3-CARBOXYLATE HYDROCHLORIDE CYCLOPENTYLACETONE

4-ISOPROPYLCYCLOHEXANONE

(+/-)-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-

7-CARBOXYLIC ACID ETHYL 2-OXO-CYCLOPENTYLACETATE ETHYL-6- (2-OXOCYCLOPENTYL) -HEXANOATE DIETHYL (2-OXOPROPYL) PHOSPHONATE MAYBRIDGE BTBG 0108 4-ACETYLPIPERIDINE HYDROCHLORIDE 4-CYANO-4-PHENYLCYCLOHEXANONE 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 3-ACETYLHEXANONE-2 l-PHENYL-2 , 4-PENTANEDIONE 3-HEXADECANONE 5-UNDECANONE 2-METHYL-3-DECANONE 4-UNDECANONE 6-TRIDECANONE 5-TRIDECANONE

1,1, l-TRIFLUORO-2 , 4-HEXANEDIONE 2-CYCLOPENTYLCYCLOPENTANONE 2-SEC-BUTYLCYCLOHEXANONE 2-BENZYLCYCLOHEXANONE 5-ACETYLVALERIC ACID 4-NITROPHENYL ACETONE 6-HEXADECANONE 5-HEXADECANONE 6-TETRADECANONE 1-HEXEN-3-ONE

2-CHLORO-6-FLUOROBENZYLIDENEACETONE MAYBRIDGE CD 09843

3-CHLORO-5,5-DIMETHYL-2-CYCLOHEXEN-l-ONE 2-ACETOXY-3-BUTANONE 5-PHENYLCYCLOHEXANE-1 , 3-DIONE TERT. -BUTYL-4-CHLOROACETOACETATE ISOPROPYL 4-CHLOROACETOACETATE 3-METHYLENE-2 , 6-HEPTANEDIONE ( 4-METHYLPHENYLTHIO) ACETONE l-(THIEN-2-YL)BUT-l-EN-3-ONE 2 -PENTADECANONE

ACETOACETOXYETHYL METHACRYLATE 4-PROPYLCYCLOHEXANONE 4-HYDROXY-2 -BUTANONE METHYL TRANS-4-OXO-2-PENTENOATE METHYL TRANS-4-OXO-2 -PENTENOATE ISOBUTYL ACETOACETATE 2 -N-HEXYLCYCLOPENTANONE 3-FLUOROPHENYLACETONE BENZYL LEVULINATE (R) - ( + ) -3 -METHYLCYCLOHEXANONE (R)-(+)-3-METHYLCYCLOPENTANONE TRANS-1-DECALONE

(S) -( + ) -10-METHYL-K9) -0CTAL-2-0NE (IS) -(-) -VERBENONE DEHYDROACETIC ACID PINONIC ACID PINONIC ACID

1 , 3 -DIAMINOACETONE DIHYDROCHLORIDE MONOHYDRATE (5S) -5, 6-ISOPROPYLIDENEDIOXY-6-METHYL-HEPTAN-2- ONE

N-OCTYL 4-CHLOROACETOACETATE MAYBRIDGE KM 02248 BICYCLO[3.2.1]OCTAN-2-ONE 2 -ADAMANTANONE BICYCLO (3.3.1) NONAN-9-ONE BICYCLO (3.3.1) NONANE-3 , 7 -DIONE (IR) -(+) -NOPINONE NORCAMPHOR NORCAMPHOR NERYLACETONE

4,4-DIMETHYL-2-CYCLOPENTEN-l-ONE 4, 4-DIPHENYL-2-CYCLOHEXEN-1-ONE THIOTETRONIC ACID

4 , 4-DIMETHYL-1 , 3 -CYCLOHEXANEDIONE 2 -CHLORO-1 , 4-BENZOQUINONE 7-OXOOCTANOIC ACID METHYL 3 -OXO-6-OCTENOATE

5 , 5-DIMETHY -2 -PHENACYL-1 , 3 -CYCLOHEXANEDIONE 5-OXOAZELAIC ACID

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

1-ONE ABSCISIC ACID ABSCISIC ACID N-CARBETHOXY-4-TROPINONE 2-METHYLTETRAHYDROTHIOPHEN-3 -ONE CYCLOHEXIMIDE PSEUDOPELLETIERINE HYDROCHLORIDE DEHYDROCARNITINE HYDROCHLORIDE

3- (PHENYLAMINO) -CYCLOHEX-2-ENE-l-ONE

3 -HEPTADECANONE

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 4-HYDROXY-3 -METHOXYPHENYLACETONE 5 , 7 -DIMETHYL-3 ,5,9-DECATRIEN-2 -ONE

ETHYL 6-METHYL-2 -OXO-3 -CYCLOHEXENE-1-CARBOXYLATE 3 -CHLOROTETRONIC ACID

2,4-DIHYDRO-5-METHYL-2-PHENYL-4-PROPIONYL-3H- PYRAZOL-3-ONE 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

3-QUINUCLIDINONE HYDROCHLORIDE 1 , 5-DIAMINO-3-OXAPENTANE 1-DIMETHYLAMINO-BUT-1-EN-3 -ONE (1R,3S) -2,2-DIMETHYL-3-(2-OXOPROPYL) - CYCLOPROPANEACETONITRILE (IS, 3S) -3-ACETYL-2, 2 -DIMETHYLCYCLOBUTANE

ACETONITRILE 5 , 5-DIMETHYLHEXANE-2 , 4-DIONE 3 -ISOBUTOXY-2 -CYCLOHEXEN-1-ONE

3 -METHYL-5-METHOXYCARBONYL-1-BENZYL-4-PIPERIDONE HYDROCHLORIDE

3-METHYL-5-METHOXYCARBONYL-4-PIPERIDONE HYDROCHLORIDE

DL-3- (1-ACETOXY-l-METHYLETHYL) -6- OXOHEPTANENITRILE

DL-3- (1-METHYL-1-ETHENYL) -6-OXOHEPTANENITRILE ETHYL 2-OXO-1-CYCLOOCTANECARBOXYLATE METHYL (lR,3S)-2,2-DIMETHYL-3- (2 -0X0PROPYL) -

CYCLOPROPANEACETATE N-BENZYLPSEUDOPELLETIERINE D-(-) -FRUCTOSE (S) - ( +) -ERYTHRULOSE HYDRATE 2,2,6, 6-TETRAMETHYL-4-PIPERIDONE MONOHYDRATE 1-BENZYL-3-PIPERID0NE HYDROCHLORIDE HYDRATE 5-ISOPROPYL-l, 3 -CYCLOHEXANEDIONE HYDRATE D-(+) -SORBOSE L-(-) -SORBOSE METHYL JASMONATE N-BENZYLTROPINONE

N-TERT-BUTOXYCARBONYL-4-PIPERIDONE (+/-) -BICYCLO(3.3.DNONANE-2, 6-DIONE ANTI-5-CARBOXYTRICYCLO[2.2.1.0 (2, 6) ] HEPTAN-3-ONE EXO-2-CHLORO-SYN-7-HYDROXYMETHYL-5-OXO- BICYCLO [2.2.1] HEPTANE L-RIBULOSE HYDRATE

4-ACETYL-2,4-DIHYDRO-5-METHYL-2-PHENYL-3H- PYRAZOL-3 -ONE MONOHYDRATE 2 -ACETYL-5-NORBORNENE

7-SYN-METHOXYMETHYL-5-NORBORNEN-2-ONE 3 -CHLORO-2 -NORBORNANONE TRICYCLO[5.2.1.02, 6] DECAN-8-ONE METHYL 4-METHOXY-2 -OXO-3 -CYCLOPENTENE-1- CARBOXYLATE

5, 5-DIMETHYL-3- (METHYLAMINO) -2-CYCLOHEXEN-l-ONE 2 -ACETYL-1 , 3 -CYCLOPENTANEDIONE 2 , 4-DIMETHOXYPHENYLACETONE 2 , 6-DIPHENYLCYCLOHEXANONE 5-HYDROXY-2-ADAMANTANONE 3 -METHOXY-2 -CYCLOPENTEN-1-ONE 2,2' -METHYLENEBIS (1,3 -CYCLOHEXANEDIONE) ( +/- ) -7-OXABICYCLO (4.1.0) HEPTAN-2-ONE ETHYL 2 -OXOCYCLOTRIDECANECARBOXYLATE 2- (METHYLTHIO) CYCLOHEXANONE

(S) - ( + ) -3 , 4, 8, 8A-TETRAHYDRO-8A-METHYL-1, 6 (2H, 7H) ^■ NAPHTHALENEDIONE, and the like.

Where aldehydes and ketones are utilized as the electrophilic reagent, the resulting 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 .

FmocH

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 [ψCH₂S] -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) 38:1:1 TFA:H2θ:Et₃SiH for 2h, lyophilize, wash (ether, 2x) then 2eq of diisopropylethylamine in 1:1 CH₃CN:H2θ, 24h. Macrocyclization kinetics of 7 is accelerated 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 B_n is benzyl) .

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 other(s) 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 e novo lead generation where the active shape ( s ) and conformation (s) 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 and 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 plate 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 in 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, Bacteriocidal/Bacteriostatic, Fungicidal/fungistatic, and the like; and

Add, Incubate, & Read assays:

Scintillation Proximity Assays, Glucan Synthase (GS) inhibition assays, Angiotensin II IPA receptor binding assay, Endothelia converting enzyme [¹²⁵I] SPA assay, HIV proteinase [1²⁵I] SPA 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

N- -Fmoc-N-ε-Boc-L-Lvs- [ψCH Sl -Glv (27)

Referring to the Method A Scheme shown above (boldface compound numberals correspond to bold-face compound numberals referred to below) , in a one liter 3-necked flask fitted with a magnetic stir bar, 22.65 grams (92mmol) of commercially available N-ε-tert- butoxycarbonyl-L-lysine 22A and 11.66 grams of sodium carbonate (llOmmol) were added to 400ml of water and stirred to give a suspension. Addition of 400ml 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 (11.72ml, llOmmol) 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 IN NaOH, then washed twice with 800ml portions of ethyl acetate (EtOAc) . The aqueous layer was acidified to pH ~3 with IN sodium bisulfate (NaHSθ4) whereupon a heavy slurry formed. The slurry was extracted twice into 800ml portions of EtOAc, then the organic layers were combined, washed with saturated brine solution, dried over sodium sulfate (Na2Sθ4) and concentrated in vacuo to produce

35grams (>100%) of 23A, a viscous pale yellow oil. TLC analysis (50:50:1 hexanes :EtOAc : cetic acid) showed clean product and this was carried to the next step assuming quantitative yield. The crude mass of 23A was dissolved in 150ml ethylene glycol dimethyl ether (DME) , chilled to -15°C (ice/water/NaCl slurry) with stirring, then 9.3 grams (10.11ml) of N-methylmorpholine (NMM) was added and the solution re-chilled. Isobutyryl chloroformate (12.56 grams, 11.93ml) 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 erlen eyer flask, retentate washed 5 times with 25ml portions of DME and the combined filtrate and rinses were re-chilled to -15°C prior to portion-wise addition of a solution 5.22 grams (138mmol, 1.5eq) sodium borohydride (NaBH4) in 150ml 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 CH₂CI₂ :EtOAc) revealed good conversion to the alcohol. The emulsion was made basic with a few ml IN NaOH, then extracted three times with 600ml portions of EtOAc. The organic layers were combined, washed with saturated brine, then dried over Na2Sθ4 and concentrated in vacuo to yield 25 grams (86%) of crude product 24A as a pale yellow oil.

The product was purified on a 500ml silica column packed and eluted with 7:3 CH2CI₂ :EtOAc to afford 17.5 grams of a nearly colorless oil 24A, 60% overall isolated yield for alloc protection and acid reduction steps . In a 250ml roundbottom flask, a portion of pure 24A (7.65 grams, 24mmol) was dissolved in 100ml dry THF under a nitrogen atmosphere, chilled on an ice bath then 5.06ml triethylamine (3.67 grams, 36mmol) was added via syringe. The solution was stirred for a few minutes on ice, then 2.06ml (3.05 grams, 26mmol) 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 CH₂CI₂ : EtOAc) indicated complete consumption of starting alcohol. The slurry was filtered, retentate washed 3 times with 100ml 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 (NaHCθ₃) and once with saturated brine before drying over Na₂S04 and concentration m. vacuo . High vacuum drying of the residue produced 9.22 grams (96%) of 25A as a colorless oil.

In a 250ml flame-dried flask fitted with a magnetic stir bar, 1.28 grams (0.97ml) of mercaptoacetic acid was dissolved in 30ml of dry dimethylformamide (DMF) under nitrogen atmosphere and to the mercapto acid was added via syringe 6.4ml (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 15ml 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 hexanes : EtOAc : acetic acid) showed 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 200ml water with a few ml of IN NaOH, and the resulting turbid solution was washed 3 times with 150ml portions of ethyl ether. 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 200ml portions of EtOAc. The combined EtOAc extracts were washed with saturated brine, dried over Na2S0₄, then concentrated to afford 4.9 grams 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 250ml flame- dried flask fitted with stir bar was added 60ml dry 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-l, 3- cyclohexanedione (dimedone) , 787 milligrams (0.25eq)of solid triphenylphosphine (PPh₃) and 693 milligrams (0.05eq) of solid tetrakis (triphenylphosphine) palladium(O) (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 50ml EtOAc and extracted 4 times with 25ml portions of water. To the combined aqueous layers was then added 2 grams solid sodium carbonate and 100ml of dioxane.

The resulting light emulsion 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 IN NaOH to the mixture and washed 3 times with 200ml portions of ethyl ether. The aqueous layer was then acidified to pH ~2 with IN NaHS04 and the off-white slurry was extracted 3 times with 150ml portions of EtOAc. The EtOAc layers were combined, washed with saturated brine, dried over N 2S04, then concentrated in vacuo to provide a yellow oil contaminated with dimedone. The oil was applied to a 500ml 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

Referring to the Method B Scheme shown above (boldface compound numerals correspond to bold-face compound numerals referred to below) , in a one liter 3 -necked flask, 11.51 grams (lOOmmol) of ±3-piperidinemethanol (28) and 10.6 grams (lOOmmol) sodium carbonate were dissolved in 200ml of water, then 200ml of dioxane was added and the mixture stirred to produce a pale yellow light emulsion. To the stirring emulsion was next added 24 grams (llOmmol) of di-tert-butyl-dicarbonate and stirring continued overnight. TLC (75:25 CH₂CI₂ : EtOAc) revealed complete conversion, so the light yellow slurry was stripped of dioxane, then extracted twice with 300ml 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 (93mmol) of 29 under nitrogen in a flame-dried one liter flask equipped with a stir bar was added 400ml dry THF via cannula. The resulting pale yellow-tan solution was chilled on an ice-water bath, then 14.12 grams (19.44ml, 139mmol) of triethylamine was added by pipette and the solution stirred for a few minutes under nitrogen. Next, 11.72 grams (7.92ml, 102mmol) 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 100ml portions of EtOAc. The combined filtrate and rinses were then washed once each with cold water, cold dilute hydrochloric acid, saturated NaHCθ₃, and saturated brine. The organic layer was dried over Na₂S04 and concentrated in vacuo to provide 26.94 grams (98%) of 30 as a pale yellow crystalline solid.

2-Mercaptonicotinic acid (11 grams, 71mmol) was dissolved under nitrogen in 400ml 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 (32.5ml, 142mmol) of a 25 weight per cent solution of NaOMe in MeOH was added to the stirring mercaptonicotinic acid/DMF solution, then 20.8 grams (71mmol) of 30 dissolved in DMF (52ml 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 400ml of water. The turbid yellow aqueous solution was partially clarified by 3 washes with 300ml portions of ethyl ether. The partially clarified aqueous layer was acidified to pH ~2 with IN NaHSθ₄ to produce a heavy yellow slurry that was then extracted 3 times with 300ml portions of EtOAc. The organic layers were combined, washed with saturated brine, dried over Na2S0₄, and concentrated in vacuo 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 (83% isolated yield) . Purified acid 31 (19 grams, 54mmol) was combined with 108ml of a 4N hydrochloric acid solution in dioxane and stirred in a one liter flask to produce a heavy white-caked precipitate. 100ml 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 HCl was removed under light vacuum on a rotovap, then the remains were neutralized with IN NaOH solution. To the neutralized solution was then added with stirring 250ml of water, 5.72 grams (54mmol) of solid sodium carbonate and another 100ml of dioxane. The resulting emulsion was chilled on an ice-water bath, then 19.12 grams (56.7mmol) 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. then the mixture was washed 3 times with 300ml portions of ethyl ether. The aqueous layer was then acidified to pH ~2 with IN NaHSθ₄ and the resulting heavy slurry extracted 3 times with 300ml portions of EtOAc. The combined EtOAc layers were washed with saturated brine, dried over Na₂Sθ4, 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

33 34 35

37 36

N-α-Fmoc-O-benzyl-L-Ser- [ψ-CH?S1 -DL-Alanine (37)

Referring to the Method C Scheme shown above (boldface compound numerals correspond to bold-face compound numerals referred to below) , in a 500ml round-bottomed flask fitted with a magnetic stir bar, 25 grams (84.6 mmol) of N-α-Boc-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/ice 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 (140-mmol, 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 CH₂CI₂ : EtOAc) revealed good conversion to the alcohol. Reaction was quenched with 1000 ml of water and the resulting emulsion was made basic with a few ml IN NaOH, then extracted three times with 300ml portions of EtOAc. The organic layers were combined, washed with saturated brine, then dried over Na₂Sθ₄ 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 1000ml 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 MsCl addition when TLC (3:1 CH2CI2 : EtOAc) 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 (NaHCθ₃) and once with saturated brine before drying over Na₂S0₄ and concentration . vacuo . High vacuum drying of the residue produced 12.7 grams (>100%) of 35 as a colorless oil.

In a 250ml 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 dimethylformamide (DMF) under nitrogen atmosphere and to the mercapto acid was added via syringe 8.24 ml (1.94 grams, 2eq) of a 25 weight percent solution of sodium ethoxide in methanol (NaOMe/MeOH). About 5.4 grams of mesylate 35 (-15 mmol) dissolved in 35 ml 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 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 yield a grayish-white paste. The paste was dissolved in 200ml water with a few ml of IN NaOH, and the resulting turbid solution was washed 3 times with 150ml portions of ethyl ether. The partially clarified aqueous layer was then acidified to pH -3 with NaHSθ₄ to yield a heavy white slurry that was extracted 3 times with 200ml portions of EtOAc. The combined EtOAc extracts were washed with saturated brine, dried over

Na₂Sθ₄, 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 : acetic acid) indicated complete conversion of starting material to a baseline product within one hour of HCl addition. Most of the HCl 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 rn vacuo, the mixture was diluted with 200 ml and then washed 3 times with 200ml portions of ethyl ether. The aqueous layer was then acidified to pH -2 with IN NaHS0₄ and the resulting slurry extracted 3 times with 100ml portions of EtOAc. The combined EtOAc layers were washed with saturated brine, dried over a₂S04, 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.

Preparation 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.

NHFmo ^■cΥ"

NHFmo ^■cΥ"

Example 1

Method for Preparation of Libraries of Macrocyclic Pseudopeptides and Organopeptides

FmocHI

e

Cvclo - (Cvs-Tyr-2-Nic rψCHSl -Phe-D-Ala-α-acetate- rψCH_ZS1 )-(!!) lOOmg (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 (β) were performed robotically at room temperature according to a custom program. The resin was then washed twice with 1.5ml of N-methyl-pyrrolidinone (NMP) . In Step (a) , the Fmoc protecting group was removed by agitating the resin with 1 ml of 30% 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 1.5ml portions of NMP. To the washed resin was then added 300uL of NMP followed by 1ml 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 250ul 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 1ml 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-

[ψCH2S] -2-mercaptonicotinate and 0.5M HOBT in NMP to give compound 8

Compound 9 was produced in step ( ) 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 α-bromoacetic acid in NMP 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 1.5ml portions of NMP, then 5 times with 1.5ml 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 2.5ml cleavage cocktail (38:1:1 trifluoroacetic acid(TFA) :H2θ: triethylsilane (Et₃SiH) or 19:1 TFA:H20) for 1.5 hours at room temperature. Another 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 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 20ml 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 centrifugation run. Ether was decanted and the pellet allowed to air dry, at which time the white to off-white solid was dissolved in 10ml 1:1 CH₃CN:H2θ, filtered through a 0.45uM teflon filter disc, then lyophilized from a tared scintillation vial. The crude material was then cyclized by dissolution in 10ml

1:1 CH₃CN:H₂0 plus lOOuL of 1M DIPEA in CH₃CN.

Other bases that promoted efficient cyclization were 2,6-lutidine and proton sponge ([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-80%) having an MH+ of 502.2, consistent with the calculated mass for 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.

38 39

Example 2

Methods for Constructing Tailed Macrocyclic

Pseudopeptides and Organopeptides .

According to a general method for constructing resins bearing both free amine group (s) 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 75ml of 1:1 dichloromethane (DCM or CH₂CI2 :N-methylpyrrolidinone (NMP), then combined with a 100ml solution containing 429mmols (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 200ml each of DCM, NMP and methanol. The wash cycle (DCM, NMP and methanol) is repeated twice 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: 0.81mmol/gram of resin) as well as both p- and m-xylenediamine-N-carbamoyl-Wang resins (resins PX and MX, respectively at 0.78mmol/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.

General Scheme for Appending Diamines and Higher Order Amines fo pNP-Wang Resin

N-p-xylenediamιno-Cyclo-(Cys-Phe-Gly[ψCH₂S]-Phe-Arg-α- acetate- [ψCH₂S] ) (18, Scheme IV)

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[ψCH2S]-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 HOBT; f) 38:1 :1 TFA:H2θ:Et3SiH for 24 hr., lyophilize, then 2 eq of diisopropylethylamine in 1 :1 CH3CN:H2θ.

With respect to Scheme IV (bold-face compound numerals and lettered steps correspond to bold-face compound numerals and lettered steps referred to below) , 65mg (50micromoles ) of PX 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 IV, steps (a) through (e) were performed robotically at room temperature according to a cumtom program written by the bench chemist. The resin was

-121-

washed and drained twice with 1.5ml of N-methyl- pyrrolidinone (NMP) . In Step (a) , to washed resin was then added 300uL of NMP followed by 1ml 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 250ul 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 1ml 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 1ml portion of piperidine/DMF solution was added, agitated for 10 minutes, then drained. Resin was then washed and drained 5 to 6 times with 1.5ml portions of NMP. To washed resin was then added in succession 300uL of NMP, 1ml of a solution 0.25M each in N-Fmoc-L-

Phenylalanine and HOBT and 250ul 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 1ml 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- [ψCH2S] -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 - 122 -

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

1.5ml portions of NMP, then 5 times with 1.5ml 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 2.5ml cleavage cocktail (38:1:1 trifluoroacetic acid (TFA) :H₂θ: triethylsilane (Et₃SiH) or 19:1 TFA:H20) for 1.5 hours at room temperature. Another 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 twice with 2.5 ml portions of water and the combined filtrate and washes were frozen and lyophilized to yield a yellow amorphous solid. Workup and cyclization were performed as in Example 1 above. Cyclization was allowed 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 used 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. -123 -

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 .

-124-

-125-

-126-

NH

H ^S_N

UeX ' HaN^N' H0^/N* H

-127-

-128-

-129-

-130-

-131-

Claims

-132 -I claim :

1. A library of structurally related macrocyclic compounds of the formula

wherein

L is a divalent group of the formula ^~ (CH₂) o-╬╣-C₆H₄CH₂- or -CHRa~ wherein R_a is hydrogen or an organic moiety;

R_z 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

wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and Ry" 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 -133-

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

- (AAl) _al-(AA2)a2 -(ATAl) tl-(AA3)_a3 -(ATA2)t2 -(AA4) _a4- (ATA3 ) _t3- (AA₅ ) _a5^~ ( A6 ) a6^~

wherein each of AAi-6 is independently a divalent group of the formula

0

N Q C

each of ATAi-3 is independently a divalent sulfur group of the formula

wherein al, a2 , a3 , a4, a5, a6, tl, t2, and t3 are independently 0 or 1, provided that tl + 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 AAi-╬▓ and ATA╬╣_3, -134-

is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from 0, 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 0, N and S; m is 0, 1 or 2 ;

Rs 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 Γûá 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.

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

3. The library of claim 1 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

T N E

Ry. "

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 -135-

6. The library of claim 1 wherein al + a2 + a3 + a4 + a5 + a╬┤ = 1, 2 or 3.

7. The library of claim 6 wherein tl + 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 0, N and S; wherein R8 is hydrogen or an organic group;

R4, R5, aⁿ⁽3 P-6 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 Ri is hydrogen, or a non-interfering substituent; and R2 is hydrogen or an organic moiety; or

Rl taken together with R_n forms a 4- to 7-membered ring, or Ri taken together with R_s forms a 4- to 7- membered ring; or Ri taken together with R2 forms a 3- to 6-membered ring. -136-

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 ^~~ (CH₂) 0-₁-C6H₄CH₂- 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 -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and Rt is hydrogen, a solid support, or Rt is a group of the formula

N E

R ΓÇóy" wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and Ry" is hydrogen or a substituent derived from an electrophilic reagent; or Ry and Ry_" taken together with the atoms to which they are - 137 -

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

(AAl) - (ATAl) - ( A3:

wherein AAl and AA3 are independently a divalent group of the formula

0

N Q c

R ^Γûán and

ATA is a divalent sulfur group of the formula

wherein is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from the group consisting of 0, 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 0, N and S; m is 0, 1 or 2 ;

R_s is hydrogen or an organic moiety; and -138 -

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 Γûá 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

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 -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and Rt is hydrogen, a solid support, or Rt is a group of the formula

N E

Ry

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 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 0, N and S; wherein R8 is hydrogen or an organic group; -139 -

R , ^R5 , and R╬╢ 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 AAl 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 Ri is hydrogen, an organic moiety or a non- interfering substituent; and

R2 is hydrogen or an organic moiety; or Rl taken together with R_n forms a 4- to 7-membered ring, or Ri taken together with R_s forms a 4- to 7- membered ring; or Ri 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 -140-

wherein

L is a divalent group of the formula ^~ (CH₂) o-╬╣-C₆H₄CH₂ ^~ 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 -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and Rt is hydrogen, a solid support, or Rt is a group of the formula

^ΓûáT- N E

y,, wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and Ry" 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 4- 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 5- to 7-membered ring or Ry and Ry" each taken together with T and the nitrogen atom - 141 -

to which they are respectively bonded form a 5- to 6- membered ring;

Z is a divalent group of the formula

^a (AAl) _a╬╣-(AA2)a2- (ATAi) _tl-(AA3)a3- (ATA2)t2-(AA4 ) _a4- ⁽ATA3⁾t3-⁽AA5⁾a5-⁽AA6⁾a6^~

wherein each of AAχ-6 is independently a divalent group of the formula

0

N Q C

R ^ΓÇón and

each of ATA1-3 is independently a divalent sulfur group of the formula

wherein al, a2 , a3 , a4, a5, a6, tl, t2, and t3 are independently 0 or 1, provided that tl + 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 AAi-6 and ATA1-3, is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from 0, N and S;

Q and OJ are each independently an organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from 0, N and S; -142-

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 ; wherein said process comprises the steps of (a) covalently bonding an amino-protected, thiol- protected starting material of the formula

NHP

wherein R_z is hydrogen or methyl, and P_s 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

-N Q COOH

Rn - 143 -

or an amino-protected thioether acid of the formula

wherein in said formulas P is an amino-protecting group and, R_n, Rn ' , Q> Q ' . ^Rs, 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

1

L COOH

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 C IIOΓÇöCH₂-C₆H₄-0-CH₂-C₆H-Polymer - 144 -

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 amino-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

O

HN T N C IIO ΓÇö CH₂ - C₆H4-0 - CH₂~ C₆ H4- Polymer

Ry Ry"

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 -145-

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 -146-

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 from the group consisting of in vi tro 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 ^~ (CH₂) o-╬╣-C₆HCH- 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 -NRyRt wherein Ry is hydrogen, alkyl, aryl or heterocycle, and - 147 -

Rt is hydrogen, a solid support, or Rt is a group of the formula

^ΓûáN

wherein E is hydrogen, a solid support or a substituent derived from an electrophilic reagent, T is a divalent linking group, and Ry" 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 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 (}AAl⁾ _a╬╣-⁽AA₂ ⁾a2- ⁽ATAl⁾ tl-⁽AA₃ ⁾a3- ⁽ATA2⁾t2-⁽AA ⁾ a4- ⁽ATA3⁾t3-⁽AA5⁾a5-⁽AA6⁾a6^"

wherein each of AA╬╣_╬▓ is independently a divalent group of the formula

0

N Q c

R ^Γûá.n and

each of ATA1-3 is independently a divalent sulfur group of the formula -148-

wherein al, a2 , a3 , a4, a5, a6, tl, t2, and t3 are independently 0 or 1, provided that tl + 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 AAi-6 and ATA1-3,

W is a divalent organic group comprising 1 to 12 carbon atoms and 0 to 4 heteroatoms selected from 0, 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 0, N and S; in 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

Rn¹ or Rs 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. - 149 -

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