Pharmacophore recombination for the identification of small molecule drug lead compounds

Abstract

A library of candidate target binding fragments (CTBF's) each CTBF being a small organic molecule and having a linkable functional group (LFG) or blocked form thereof (BLFG), wherein the LFG or BLFG contains a linking group (LG), the LG being a disulfide group.

What is claimed is:  
     
         1 . A method for identifying a drug lead compound that binds to a target biological molecule (TBM) of interest, the method comprising: 
 (a) assembling a library of candidate target binding fragments (CTBF) capable of being chemically cross-linked by a cross-linker to provide candidate cross-linked target binding fragments for binding to the target biological molecule;    (b) screening the library of candidate target binding fragments to identify at least first and second candidate target binding fragments that bind to the target biological molecule;    (c) chemically cross-linking the at least first and second candidate target binding fragments or structurally related analogs thereof with a cross-linker to provide a library of candidate cross-linked target binding fragments for binding to the target biological molecule; and    (d) screening the library obtained in (c) to identify a drug lead compound that binds to the target biological molecule.    
     
     
         2 . The method according to  claim 1 , wherein at least one of the candidate target binding fragments of the library of candidate target binding fragments binds to the target biological molecule with a K d  of from about 5 mM to about 0.05 mM.  
     
     
         3 . The method according to  claim 1 , wherein at least one of the candidate target binding fragments of the library of candidate target binding fragments binds to the target biological molecule with a K d  of from about 3 mM to about 0.1 mM.  
     
     
         4 . The method according to  claim 1 , wherein the drug lead compound identified in step (d) binds to the target biological molecule with a K d  of 500 nM or lower.  
     
     
         5 . The method according to  claim 1 , wherein the screening steps (b) and (c) consist essentially of an in vitro biological assay.  
     
     
         6 . The method according to  claim 1 , wherein the library of candidate cross-linked target binding fragments for binding to the target biological molecule comprises homodimeric or heterodimeric candidate cross-linked target binding fragments.  
     
     
         7 . The method according to  claim 1 , wherein the library of candidate target binding fragments comprises candidate target binding fragments of less than about 500 daltons.  
     
     
         8 . The method according to  claim 1  wherein the library of candidate cross-linked target binding fragments comprises candidate cross-linked target binding fragments of less than about 750 daltons.  
     
     
         9 . The method according to  claim 1 , wherein the target biological molecule is a human or human pathogen protein.  
     
     
         10 . The method according to  claim 9 , wherein the protein is an enzyme, a human hormone, a human receptor and fragments thereof having nitrogen's in the protein present in their naturally occurring isotopic abundance.  
     
     
         11 . The method according to  claim 1 , wherein at least one of the screening steps (b) and (d) is accomplished by ELISA assay.  
     
     
         12 . A method for inhibiting the binding of a first biological molecule to a second biological molecule that binds to the first biological molecule, the method comprising: 
 contacting a system comprising both the first and second biological molecules with a binding inhibitory amount of a drug lead compound identified according to the method of  claim 1 , wherein the drug lead compound binds to the first biological molecule and inhibits its ability to bind to the second biological molecule.    
     
     
         13 . The method according to  claim 12 , wherein the first and second biological molecules are human proteins.  
     
     
         14 . The method according to  claim 13 , wherein the first and second biological molecules are selected from the group; a human hormone, cytokine or chemokine, a human receptor and fragments thereof  
     
     
         15 . A method for identifying a drug lead compound that binds to a target biological molecule of interest, the method comprising: 
 (a) assenbling a library of candidate target binding fragments, each fragment containing an oxime linking group;    (b) screening the library of candidate target binding fragments or monomers to identify at least first and second oxime containing candidate target binding fragments that bind to the target biological molecule;    (c) chemically crosslinking the aldehyde analogs of the at least first and second oxime containing candidate target binding fragments with an O,O′-diamino-alkanediol cross-linker to provide a library of oxime containing candidate cross-linked target binding fragments for binding to the target biological molecule; and    (d) screening the library obtained in (c) to identify a drug lead compound that binds to the target biological molecule.    
     
     
         16 . A method comprising: 
 (a) assembling a library of candidate target binding fragments (CTBF), each fragment having a linkable functional group (LFG) or blocked form thereof (BLFG), the blocked form containing a linking group (LG);    (b) contacting the candidate target binding fragments with a target biological molecule (TBM);    (c) measuring a change in a first physical association (PA-1) of the target biological molecule;    (d) selecting target binding fragments (TBF) based on (c);    (e) reacting selected target binding fragments having a linkable functional group with a cross-linker, having chemically compatible cross-reactive groups (CFG) with the LFG, under conditions suitable for forming a library of candidate cross-linked target binding-fragments (CXL-TBF);    (f) contacting the candidate cross-linked target binding fragments with the target biological molecule (TBM);    (g) measuring a change in a second physical association (PA-2) of the target biological molecule;    (h) selecting cross-linked target binding fragments (XL-TBF) based on (g).    
     
     
         17 . The method of  claim 16  wherein the candidate target binding fragment contacted with the TBM contains a blocked linkage functional group (BLFG) containing linking group LG.  
     
     
         18 . The method of  claim 17  where in the linking group (LG) in BLFG is selected from the group; oxime, hydrazone, N-acyl hydrazone, secondary amine, tertiary amine, acetal, ketal, 1,2 amino alcohols, amide, N,N-disubstituted amides, thioamide, ureido, thioureido, carbamate, thiocarbamate, thiothiocarbamate, sulfonamide, carbonate, guanidino, amidino, thioester, ester, ether, 2-hydroxyether, 2-hydroxythioether, thioether, disulfide, alkane (alkylene), alkene (alkenylene) and alkyne (alkynylene).  
     
     
         19 . The method of  claim 18  wherein each candidate target binding fragment (CTBF) of step (b) contains the same linking group (LG) as is present in the candidate cross-linked target binding fragment (CXL-TBF) of step (f).  
     
     
         20 . The method of  claim 19  wherein the linking group (LG) is selected from the group; oxime, secondary amine, tertiary amine, amide, ureido, thioureido, sulfonamide and carbamate.  
     
     
         21 . The method of  claim 19  wherein two candidate target binding fragments (CTBF) selected from step (d) are cross-linked to form a candidate cross-linked target binding fragment (CXL-TBF) represented by  
       
         
           
           
               
               
           
         
       
       where 
 TBF m  is a first TBF selected from step (d) which contained LG, in its blocked linking group;  
 TBF n  is a second TBF selected from step (d) which contained LG 2  in its blocked linking group;  
 XL represents the cross-linker without the chemically compatible cross-reactive functional groups;  
 LG 1  represents the linking group in the first TBF; and  
 LG 2  represents the linking group in the second TBF.  
 
     
     
         22 . The method of  claim 21  wherein LG 1  and LG 2  are the same or are different and are selected from the group oxime, secondary amine, tertiary amine, amide, ureido, thioureido, sulfonamide and carbamate.  
     
     
         23 . The method of  claim 21  wherein TBF m  and TBF n  are the same or are different.  
     
     
         23 . The method of  claim 21  where XL is selected from the group of alkanes: methylene, ethylene, propylene, butylene, pentylene, hexylene and heptylene,optionally containing 0, 1, 2 or 3 ether linkages and from 1-3 double bonds and aryls: ortho-, meta- or para- C 0 -C 6 -alkyl-phenyl-C 0 -C 6 -alkylene.  
     
     
         24 . The method of  claim 16  wherein the candidate cross-linked target binding fragments are represented by the formula:  
       
         
           
           
               
               
           
         
         where  
         TBF m  represents a first TBF selected from step (d);  
         TBF n  represents a second TBF selected from step (d);  
       
       XL represents the cross-linker without the chemically compatible cross-reactive functional groups selected from the group 
 C 0 —C 10 ,-alkylene,  
 C 0—C   6 -alkyl-C 6 —C 10 -aryl-C 0 —C 6 -alkylene,  
 C 1 —C 6 -alkyl-N(R 1 )—C 1 —C 6 -alkylene,  
 (C 1 —C 6 -alkyl-O—C 1 —C 6 -alkylene) n , where n═1,2,3 or 4;  
 LG 1  and LG 2  are linking groups independently selected from the group  
 —C(R a )═N—O—, —O—N═C(R a )—, —CH 2 —N(R a )—, —N(R a )—CH 2 —,  
 —C(═O)—N(R a )—, —N(R a )—C(═O)—, —N(R a )—C(═O)—O—, —O—C(═O)—N(R a )—,  
 —N(R a )—C(═O)—N(R b )—, —N(R a )—C(═O)—N(R b )—, —SO 2 —N(R a )— and  
 —N(R a )—SO 2 —;  
 R a  and R b  are independently selected from the group 
 hydrogen, C 1 —C 10 -alkyl, C 0 —C 10 -alkyl-C 6 —C 10 -aryl, C 6 —C 10 -aryl-C 0 —C 10 -alkyl, 
 C 0 —C 10 -alkyl-heterocycle-C 0 —C 10 -alkyl, C 1 —C 6 -alkyl-NH-C 1 —C 6 -alkyl,  
 C 0 —C 10 -alkyl-O—C 0 —C 10 -alkyl, C 0 —C 10 alkyl-C(═O)-C 0   —C   10 -alkyl, C 0 —C 10 -alkyl-NH—C(═O)—C 0 —C 10 -alkyl,  
 C 0 —C 10 —alkyl-O—C(═O)—C 0 —C 10 -alkyl,  
 where any alkyl, aryl or heterocycle is optionally substituted with C 1 —C 10 —alkyl, C 1 —C 10 -alkoxy, C 6 —C 10 -aryl, C 6 —C 10 -aryloxy, halo(F, Cl, Br, I), hydroxy, carboxy, amino, nitro and S(O) 0−3 .  
 
     
     
         25 . The method of  claim 24  wherein the TBF m  and TBF, from step (d) each independently bind to the target biological molecule with a K d  of from about 3 mM to about 100 μM.  
     
     
         26 . The method of  claim 25  wherein the TBF m  and TBF n  from step (d) each independently bind to the target biological molecule with a K d  of from about 2 mM to about 500 μM.  
     
     
         27 . The method according to  claim 24 , wherein the target biological molecule is a human or human pathogen protein.  
     
     
         28 . The method according to  claim 24 , wherein the protein is an enzyme, a human hormone or a human receptor having nitrogen's in their naturally occuring isotopic abundance.  
     
     
         29 . The method according to  claim 24 , wherein at least one of steps (b) and (d) is accomplished by ELISA assay.  
     
     
         30 . The method of  claim 21  wherein  
       
         
           
           
               
               
           
         
       
       is selected from the group;  
       
         
           
           
               
               
           
         
       
     
     
         31 . The method of  claim 16  wherein the candidate cross-linked target binding fragments are represented by the formulae:  
       
         
           
           
               
               
           
         
       
       where 
 TBF m  represents a first TBF selected from step (d);  
 TBF m  represents a second TBF selected from step (d);  
 TBF m -part A and B represent TBF m  from step (d) where each fragment is bonded to a single atom in LG 3 ;  
 TBF n -part C and D represent TBF n  from step (d) where each fragment is bonded to a single atom in LG 4 ;  
 XL represents a cross-linker of the formula  
 —(C 0 —C 2 -alkyl-L 1 -L 2 -L 3 -L 4 -L 5 -C 0 —C 2 -alkyl)-;  
 LG 1  and LG 2  are linking groups independently selected from the group —C(R a )═N—O—, —O—N═C(R a )—, —CH 2 —N(R a )—, —N(R a )—CH 2 —, —C(═O)—N(R a )—, —N(R a )—C(═O)—, —N(R a )—C(═O)—O—, —O—C(═O)—N(R a )—, —N(R a )—C(═O)—N(R b )—, —N(R a )—C(═O)—N(R b )—, —SO 2 —N(R a )— and —N(R a )—SO 2 —;  
 LG 3  and LG 4  are linking groups independenlty selected from the group >C═N—O—, —O—N═C<, —CH 2 —N<, >N—CH 2 —, —C(═O)—N<, >N—C(═O)—, >N—C(═O)—O—, —O—C(═O)—N<, >N—C(═O)—N(R b )—, —N(R a )—C(═O)—N<, —SO 2 —N< and >N—SO 2 —, where < and > represent two bonds linking TBF-part A, B, C, or D to the single N or C atom in LG 3  or LG 4 ;  
 R a  and R b  are independently selected from the group hydrogen, C 1 —C 10 -alkyl, C 0 —C 10 -alkyl-C 6 —C 10 -aryl, C 6 —C 10 -aryl-C 0 —C 10-alkyl, C   0 —C 10 alkyl-heterocycle-C 0 —C 10 -alkyl, C 1 —C 6 -alkyl-NH—C 1 —C 6 -alkyl, C 0 —C 10 -alkyl-O—C 0 —C 10 -alkyl, C 0 —C 10 alkyl-C(═O)-C 0—C   10-alkyl, C   0 —C 10 -alkyl-NH—C(═O)—C 0 —C 10 -alkyl, C 0 —C 10 -aklyl—O—C(═O)—C 0 —C 10 -alkyl, where any alkyl, aryl, aryl or heterocycle is optionally substituted with C 1 —C 10 -alkyl, C 1 —C 10 -alkoxy, C 6 —C 10 -aryl, C 6 —C 10 -aryloxy, halo (F, Cl, Br, I), hydroxy, carboxy, amino, nitro and S(O) 0−3 ;  
 TBF m , TBF n , TBF m -part A, TBF m -part B, TBF n -part C and TBF n -part D are each independently represented by formula I  
 -A-(Cycle 1)-B-(Cycle 2)-E  (I)  
 Where  
 Cycle  1  and Cycle  2  are independently present or absent and are selected from a mono-, bi-, or tricyclic saturated, unsaturated, or aromatic ring, each ring having 5, 6 or 7 atoms in the ring where the ring atoms are carbon or from 1-4 heteroatoms selected from; nitrogen, oxygen, and sulfur, and where any sulfur ling atom may optionally be oxidized and any carbon ring atom may form a double bond with O, NR n  and CR 1  R 1 ′, each ring nitrogen may be substituted with Rn and any ring carbon may be substituted with R d ;  
 A and B are independently selected from  
                     
 where: 
 L 1  is absent or may be selected from oxo (O), S(O) s , C(═), C(═N—R n ). C(═CR 1 R 1′ ), C(R 1 R 1′ ), C(R 1 ), C, het, N(R n ) or N;  
 L 2  is absent or may be selected from oxo (O), S(O) s , C(═O), C(═N—R n ), C(═CR 2 R 2 ′), C(R 2 R 2′ ), C(R 2 ), C, het, N(R n ) or N;  
 L 3  is absent or may be selected from oxo (O), S(O) s , C(═O), C(═N—R n ), C(═CR 3 R 3′ ), C(R 3 R 3′ ) C(R 3 ), C, het, N(R n ) or N;  
 L 4  is absent or may be selected from oxo (O), S(O) s , C(═O), C(═N—R n ), c(═CR 4 R 4′ ), C(R 4 R 4′ ), C(R 4 ), C, NR n  or N; and  
 L 5  is absent or amy be selected from oxo (O), S(O) s , C(═O), C(═N—R n ), C(R 5 R 5′ ), C(═CR 5 R 5′), C(R   5 ), C, NR n  or N;  
 R 1 , R 1′ , R 2 , R 2′ , R 3 , R 3′ , R 4 , R 4′, R   5  and R 5′  each are independently selected from R a , R a′ , R c  and U-Q-V-W; where s is 0-2  
 
 Optionally, each R 1 —R 5  or NR n  together with any other R 1 —R 5  or NR n  may form a mono-, bi, or tricyclic saturated, or aromatic ring, each ring being a homo- or hereocycle having 5, 6 or 7 atoms in the ring, optionally each ring containing 1-4 hereroatoms selected from N, O and S where any ring carbon or sulfer atom may optionally be oxidized, each ring nitrogen optionally substituted with R n  and each ring carbon optionally substituted with R d  ;  
 E is -L 1 -L 2 -L 3 -R a ;  
 R a  is selected from the group; hydrogen, halo(F, Cl, Br, I), halo(F, Cl, Br, I)—C 1 —C 11 alkyl, halo(F, Cl, Br, I)—C 1 —C 11 alkoxy, hydroxy-C 1 —C 11 alkyl, cyano, isocyanate, carboxy-C 1 —C 11 alkyl, amino, C 0 —C 11 alkyl-amino-(C 1 —C 8 alkyl), C 0 —C 11 alkyl-amino-di-(C 1 —C 8 alkyl), aminocarbonyl, C 1 —C 11 alkylcarbonylamino, carboxamido, carbamoyl, carbamoyloxy, formyl, formyloxy, azido, nitro, hydrazide, hydoxamic acid, imidazoyl, ureido, thioureido, thiocyanato, hydroxy, C 1 —C 6 alkoxy, mercapto, sulfonamido, het, phenoxy, phenyl, benzyl, benzyloxy, benzamido, tosyl, morpholino, morpholinyl, piperazinyl, piperidinyl, pyrrolinyl, imidazolyl and indolyl;  
 R a′  is selected from the group of C 0 —C 10 alkyl-Q-C 0 —C 6 alkyl, C 0 —C 10 alkenyl-Q-C 0 —C 6 alkyl, C 0 —C 10 alkynyl-Q-C 0 —C 6 alkyl, C 3 —C 11 cycloalkyl-Q-C 0 —C 6 alkyl, C 3 —C 10 cycloalkenyl-Q-C 0 —C 6 alkyl, C 1 —C 6 alkyl-C 6 —C 12 aryl-Q-C 0 C 6 alkyl, C 6 —C 10 aryl-C 1 —C 6 alkyl-Q-C 0 — 6 alkyl, C 0 —C 6 alkyl-het-Q-C 0 —Calkyl, C 0 —C 6 alkyl-Q-het-C 0 —C 6 alkyl, het-C 0 —C 6 alkyl-Q-C 0 —C 6 alkyl, C 0 —C 6 alkyl-Q-C 6 —C 12 aryl and Q-C 1 —C 6 alky, where any aryl or het is optionally with 1-3 R d  and alkyl, alkenyl or alkynyl is optionally substitued with 1-3 R a ;  
 R a  and R a′  may join to form a 3-7 member homocyclic ring substitued with 1-3 R a ;  
 R c  is selected from hydrogen and substituted or unsubstitued; amino O—C 1 —C 8 alkyl, amino-(C 1 —C 8 alkyl), amino-di-(C 1 —C 8 alkyl), C 1 —C 10 alkyl, C 2 —C 10 alkenyl, C 2 —C 10 alkynyl, C 3 — 11 cycloalkyl, C 3 —C 10 cycloalkeynl, C 1 —C 6 alkyl-C 6 —C 12 aryl, C 6 —C 10 aryl-C 1 —C 6 alkyl, C 1 —C 6 alkyl-het, het-C 1 —C 6 alkyl, C 6 —alkyl, C 6 —C 12 aryl and het, where the substituits on any alkyl, alkenyl or alkynyl are 1-3 R  a  and the substituents on any aryl or het are 1-3 R d ; 
 R d  is selected from R h  and R p;    
 R h  is selected form the group of OH, OCF 3 , OR c , SR m , halo(F, Cl. Br, I), CN isocyanate, NO 2 , CF 3 , C 0 —C 6 alkyl-NR n R n′ , C 0 —C 6 alkyl-C(═O)-NR n R n′ , C 0 —C 6 alkyl-C(═O)—R a , C 1 —C 8 alkyl, C 1 —C 8 alkoxy, C 2 —C 8 alkenyl, C 2 —C 8 alkynyl, C 3 —c 6 cycloalkyl, C 3 —C 6 cycloalkenyl, C 1 —C 6 alkyl-phenyl, phenyl-C 1 —C 6 alkyl, C 1 —C 6 alkyloxycarbonyl, phenyl-C 0 —C 6 alkyloxy, C 1 —C 6 alkyl-het, het-C 1—C   6 alkyl, SO 2 -het, O—C 6 —C 12 aryl, SO 2 —C 6—C   12 aryl, SO 2 —C 1 —C 6 alkyl and het, where any alkyl, alkenyl or alkynyl may optionally be stubstitued with 1-3 groups selected from OH, halo(F, Cl, Br, I), nitro, amino, and aminocarbonyl, where the substituents on any aryl or het are 1-2 hydroxy, halo(F, Cl, Br, I), CF 3 , C 1 —C 6 alkyl, C 1 —C 6 alkoxy, nitro and amino;  
 
 R m  is selected from hydrogen, S—C 1 —C 6 alkyl, C(═O)—C 1 —C 6 alkyl, C(═O)—NR n R n′ , C 1 —C 6 alkyl, halo(F, Cl, Br, I)—C 1 —C 6 alkyl, benzyl and phenyl;  
 R n  is selected from the group R c , OH, OCF 3 , OR o , CN, Isocyanate, NH—C(═O)—O—R 3 , NH—C(═O)—R c , NH—C(═O)—NHR c , NH—SO 2 —R s , NH—SO 2 —NH—C(═O)—R c , NH—C(═O)—NH—SO 2 —R s , C(═O)—O—R o , C(═O)—R c , C(═O)—NHR c, C(═O)—NH—C(═O)—O—R   o , C(═O)—NH—C(═o)—R c , C(═O)—NH—SO 2 —R s , C(═O)—NH—SO 2 —NHR 3 , SO 2 —R s , SO 2 —O—R o , SO 2 —N(R c ) 2 , SO 2 —NH—C(═O)—O—R o  , SO 2 —NH—C(═O)—O—R o  and SO 2 —NH—C(═O)—R c ;  
 R o  is selected from hydrogen and substituted or unsubstituted C 1 —C 6 alkyl, C 0 —C 6 alkyl—C 6 —C 10 aryl, C 1 —C 6 alkylcarbonyl, C 2 —C 6 alkenyl, C 2 —C 6 alkynyl, C 3 —C 8 cycloalkyl and benzoyl, where the substituits on any alkyl are 1-3 R a  and the substituents on any aryl are 1-3 R p ; R p  is selected from the group; OH halo(F, Cl. Br, I), CN, isocyanate, OR o , SR m , SOR o , NO 2 , CF 3 , R c , NR n R n′ , N(R n )—C(═O)—O—R o , N(R n )—C(═O)—R c , C 0—C   6 alkyl—SO 2 —R s , C 0 —C 6 alkly—SO 2 —NR n R n′ , C(═O)—R c , O—C(═O)—R c , C(═O)—O—-R o  and C(═O)—NR n R n′ , where the substituits on any alkyl, alkenyl or alkynyl are 1-3 R a  and the substituents on any aryl or het are 1-3 R d ;  
 R s  is a substituted or unsubstituted group selected form; C 1 —C 8 alky, C 2 —C 8 alkenyl, C 2 —C 8 alkynyl, C 3 —C 8 cycloalkyl, C 3 —C 6 cycloalkenyl, C 0 —C 6 alkyl-phenyl, phenyl—C 0 —C 6 alkyl, C 0 —C 6 alkyl-het and het-C 0 —C 6 alkyl, where the substituits on any alkyl, alkenyl or alkynyl are 1-3 Ra and the substituents on any aryl or het are 1-3 Rd  
 het is any mono-, bi-, or tricyclic saturated, unsaturated, or aromatic ring where at least one ring is a 5-, 6- or 7-membered ring containing from one to four heteroatoms selected from the group nitrogen, oxygen, and sulfur, the 5-membered ring having from 0 to 2 double bonds and the 6- or 7-membered ring having from 0 to 3 double bonds and where any carbon or sulfur atoms in the ring may optionally be oxidized, and where any nitrogen heteroatom may optionally be quaternized and where any ring may contain from 0-3 Rd;  
 U is an optionally substituted bivalent radical selected from the group; C 1 —C 6 alkyl, C 0 —C 6 alkyl-Q, C 2 —C 6 alkenyl-Q, and C 2 —C 6 alkynyl-Q, where the substituits on any alkyl, alkenyl or alkynyl are 1-3 R a ;  
 Q is absent or is selected for the group;—O—, S(O) s —, —SO 2 —N(R n ) 13  N(R n )—, —N(R n )—C(═O)—, —N(R n )—C(═o)—O—, —N(R n )—SO 2 —, —C(═O)—, —C(═O)—O—, -het, —C(═O)—N(E n )—, —PO(OR c )O— and —P(O)O—, where s is 0-2 and the heterocyclic ringis substitued with 0-3 R h ;  
 V is absent or is an optionally substitued bivalent group selected from C 1 —C 6 alkyl, C 3 —C 8 cycloalkyl, C 0 —C 6 alkyl-C 6 C 10 aryl, and C 0 —C 6 alky-het, where the substituits on any alkyl are 1-3 R a  and the substituents on any aryl or het are 1-3 R d ;  
 W is selected from the group; hydrogen; —OR o , —SR m , —NR n R n′ , —NH—C(═O)—O—R o —NH—C(═O)—NR n R n′ , —NH—C(═O)—R C , —NH—SO 2   13  R s , —NH—SO 2 —NR n R n′ , —NH—SO 2 —NH—C(═O)—R c , —NH—C(═O)—NH—SO 2 R s , —C(═O)—NH—C(═O)—O—R o , —(═)—NH—C(═O)—R c , —C(═O)—NH—C(═O)—NR n R n′ , —C(═O)—NH—SO 2 —R 8 , —C(═O)—NH—SO 2 —NR n R n′, —C(═s)—NR   n R n′ , —SO 2 —R s , —SO 2 —O—R o, —SO   2 —NR n R n′ , —SO 2 —NH—C(═O)—O—R o , —SO 2 —NH—C(═O)—NR n R n′ , —SO 2 —NH—C(═O)—R c , —O—c(═O)—NR n R n40  , —C(═O)—R c —O—C(═O)—NH—C(═O)—R c , —O—C(═o)—NH—SO 2 —R s  and —O—SO 2—R   s ;  
 Optionally, TBF m -part A together with TBF m -part B and TBF n -part C together with TBF n -part D may independently form (Cycle  1 ) substituted with —B-(Cycle  2 )-E.  
 
     
     
         32 . A compound made by the method of claim  30 .