United States Patent (19) 11 Patent Number: 5,766,963 Baldwin Et Al
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IIIIUS005766963A United States Patent (19) 11 Patent Number: 5,766,963 Baldwin et al. 45 Date of Patent: Jun. 16, 1998 54 COMBINATION HYDROXYPROPYLAMINE Primary Examiner-Ponnathapura Achutamurthy LIBRARY Attorney, Agent, or Firm-Heslin & Rothenberg, PC 75) Inventors: John J. Baldwin, Gwynedd Valley, Pa.; 57 ABSTRACT Ian Henderson, Plainsboro; Frank S. A combinatorial library is disclosed which is represented by Waksmunski. South River, both of N.J. Formula 73) Assignee: Pharmacopeia, Inc.. Princeton, N.J. (21) Appl. No.: 592,654 22 Filed: Jan. 26, 1996 (51) Int. Cl. .................... G01N 33/543; G01N 33/544; wherein: GO1N 33/545; GO1N 33/551 52 U.S. Cl. .......................... 436/518; 436/501; 436/523; (S) 436/524; 436/527; 436/528: 436/529; 436/530: 436/531; 435/4 is a solid support; T-L- is an identifier residue; and 58) Field of Search ................................ 435/4. 7.1, 7.21, -L-IT is a ligand/linker residue. This library contains 435/7.71, 7.8, 7.93, 7.4, 7.7, 7.9; 436/501, hydroxypropylamines of the formula: 518, 523, 524,527-531 56 References Cited wherein: U.S. PATENT DOCUMENTS Aa' and Aa is each an amino acid joined to each other 4.383,031 5/1983 Boguslaski et al. ........................ 435/7 through an amide bond; 5,525,735 6/1996 Gallop et al.............. ... 548/533 Ar" is an aromatic ring system; 5,573,905 11/1996 Lerner et al. ............................... 435/6 -CHAr" is attached to N on Aa; and OTHER PUBLICATIONS R" is H. Coalkyl, alkenyl, alkynyl, aryl, heteroaryl, aryl or heteroaryl fused to a 3- or 4-membered moiety to Brenner & Lerner, "Encoded Combinatorial Chemistry". form a non-aromatic second ring, or substituted C-2 Proc. Nat. Acad. Sci. USA 89,5381-5383 (1992). alkyl, alkenyl, or alkynyl. Burbaum et al. "A Paradigm for Drug Discovery Employing Encoded Combinatorial Libraries', Proc. Natl. Acad. Sci USA 92, 6027-6031 (1995). 6 Claims, No Drawings 5,766.963 1 2 COMBINATION HYDROXYPROPYLAMINE cannot contain a linear chain of 3, 4, or 5 atoms which LIBRARY separate the carboxyl carbonyl from the amino group of Aa", and Aa' cannot be an O-amino acid; CROSS-REFERENCE Ar' is aryl or heteroaryl; Lawn Assay for Compounds That Affect Enzyme Activity -CHAr' is attached to N on Aa’; or Bind to Target Molecules, U.S. Ser. No. 08/553.056, filed R" is H. Co alkyl, alkenyl, alkynyl, aryl, heteroaryl, Nov. 3, 1995, is incorporated herein by reference. substituted aryl or heteroaryl, aryl or heteroaryl fused All patents and other references cited herein are hereby to a 3- or 4-membered moiety to form a non-aromatic incorporated by reference. second ring, or substituted Co alkyl, alkenyl, or O alkynyl; and BACKGROUND OF THE INVENTION X is O. N-loweralkyl, S. S(O), or S(O). There is interest in methods for the synthesis of large Preferred libraries of Formula I are those wherein T-L- is numbers of diverse compounds which can be screened for of the Formula various possible physiological or other activities. Tech 15 niques have been developed in which one adds individual o-ch units sequentially as part of the chemical synthesis to O-Air produce all or a substantial number of the possible com pounds which can result from all the different choices --- OCH3 possible at each sequential stage of the synthesis. For these 20 O techniques to be successful, it is necessary for the com pounds to be amenable to methods by which one can wherein: determine the structure of the compounds so made. Brenner n=3-12; and Lerner (PNAS USA 81: 5381-83 (1992)) and WO Ar is halophenyl; and 93/20242, for example, describe a synthesis wherein oligo 25 q is 3-12. nucleotides are produced in parallel with and are chemically Other preferred libraries of Formula I are those wherein linked as genetic tags to oligopeptides as the compounds of interest. WO93/06121 teaches methods for particles-based -L"- is synthesis of random oligomers wherein identification tags (a) on the particles are used to facilitate identification of the 30 oligomer sequence synthesized. A detachable tagging sys Vxy tem is described in Ohlmeyer et al., Proc. Natl. Acad. Sci. USA, 90, 10922-10926. Dec. 1993. wherein the left-hand bond as shown is the point of SUMMARY OF THE INVENTION 35 attachment to the solid support and the right hand bond The present invention relates to a combinatorial library of is the point of attachment to the ligand. compounds encoded with tags and to the use of this library More-preferred libraries of Formula I are those wherein in in assays to discover biologically active compounds. The Formula III: 1) n=3-12 and Ar is pentachlorophenyl; or 2) present invention also relates to a library of hydroxypropy n=5-6 and Ar is 2,4,6-trichlorophenyl. lamine compounds containing two amino acid residues and 40 Depending on the choice of L' (see Table 1), the ligands a heteroatom-substituted hydroxypropyl residue and using of Formula II may be detached by photolytic, oxidative. this library to identify biologically active members by acidic, basic, or other cleavage techniques. For example, screening in bioassays. when -L"- is (a), acidic cleavage may be represented by: DETALED DESCRIPTION OF THE 45 INVENTION The combinatorial chemical library of the present inven wherein L" is the residue from L' and ITOH is II: tion is represented by Formula I: Aa'-Aa’-(CHAr")-CHCHOH-CHXR" I A preferred embodiment of the invention is a library of wherein: Formula I wherein: Aa' is selected from the seven residues of the amino acids (S) of Table 1-1; 55 Aa is selected from the 15 residues of the amino acids of is a solid support; Table 1-2; T-L- is an identifier residue; Ar' is selected from the 31 aryl and heteroaryl residues of -L-IT is a linker?ligand residue; and the aldehydes of Table 1-3; R" is selected from the 31 alkyl, aryl, arylalkyl, and het q is 2-30; and eroaryl residues of epoxides of Table 1-4; and II is X is O or S. One embodiment of the invention is the use of the combinatorial library of Formula I in assays to discover biologically active compounds (ligands) of Formula II. wherein: 65 Thus, an aspect of the invention is a method of identifying Aa' and Aa’ are each an amino acid joined to each other a compound having a desired characteristic which comprises through an amide bond with the provisos that Aa' synthesizing a combinatorial library of Formula I and testing 5,766.963 3 4 the library of Formula I, either attached to the solid support 1) reacting a compound of the formula 3: or detached therefrom, in an assay which identifies com pounds of Formula II having the desired characteristic. (S)-O-Aa-AaHFmoc 3 Another embodiment of the invention is a method of iden tifying a compound having a desired characteristic which with piperidine/DMF at room temperature for about 1 comprises testing the library of Formula I, either attached to hir: the solid support or detached therefrom, in an assay which 2) reacting the product of step 1 with a carboxaldehyde identifies compounds of Formula II having the desired reagent of formula Ar" CHO, dissolved in toluene. at characteristic. A further embodiment of the invention is room temperature for about 15 hr to produce an imine; determining the structure of any compound so identified. O and It is within the scope of the present invention that the 3) suspending the imine of step 2 in methanol with sodium determination of the structures of compounds having the cyanoborohydride at room temperature for about 4 hr to desired characteristic can be accomplished by decoding the produce a library of compounds 4. tags (represented by T-L- in Formula I) or, alternatively, by Another embodiment of the invention is the use of deconvolution of the library (Smith et al. BioMed. Chem. 5 divinylbenzene-cross-linked, polyethyleneglycol-grafted Lett, 4. 2821 (1994); Kurth et al., J. Org. Chem. 59, 5862 polystyrene beads optionally functionalized with amino (1994); Murphy et al. J. Am. Chem. Soc., 117.7029 (1995); groups (for example. TentaGel(ES NH. Rapp Polymere) as Cambell et al., J. An. Chem. Soc., 117.5381 (1995); and Erb the solid supports for constructing a combinatorial library of et al., Proc. Nat. Acad. Sci. USA, 91, 11422 (1994)). In the Formula I or I. latted case, the library of the present invention is represented 20 Definitions by Formula T The following abbreviations have the indicated meaning: (S)-CO)-L-II DEAD-diethylazodicarboxylate wherein the symbols are as defined for Formula I. 25 DCM=dichloromethane=methylene chloride Another embodiment of the invention is a method of DIC=diisopropylcarbodiimide synthesizing a library of Formula Ia which comprises react DMAP-4-N,N-dimethylaminopyridine ing a compound of Formula 4 DMF=N,N-dimethylformamide DVB-1,4-divinylbenzene 30 FACS-fluorescence activated cell sorting T \-Al Fmoc=9-fluorenylmethoxycarbonyl with an epoxide of the formula GC=gas chromatography HOBt-hydroxybenzotriazole O n-eleta 35 Me=methyl RX-1- Mtr=4-methoxy-2,3,6-trimethylbenzenesulfonyl NaBHCN=sodium cyanoborohydride dissolved in a suitable solvent such as acetonitrile or a lower PEG=polyethylene glycol alcohol, e.g. isopropanol, at 25°-82°C. to produce a library Ph=phenyl of Formula Ia 40 rt.=room temperature sat'd=Saturated (S-co-L-A-A1N1 XR (Ia) S-secondary OH t-=tertiary Arl TFA=trifluoroacetic acid 45 THF=tetrahydrofuran wherein: Thy=thienyl HO-Aa"-H=Aa" with the HO- incorporated into the Alkyl is intended to include linear, branched, or cyclic carboxyl group of the amino acid Aa" and the -H hydrocarbon structures and combinations thereof.