IL122092A - Cyclic 3, 4-bis (3-indolyl) maleic anhydride and 3, 4-bis (3-indolyl) maleimide intermediates for protein kinase c inhibitors - Google Patents

Abstract

A compound of the formula wherein: V is -O- or -N(CH3)-; W is -O-, -S-, -SO-, -SO2-, -CO-, C2-C6 alkylene, substituted alkylene, C2-C6 alkenylene, -arylene-, -arylene (CH2)mO-, -heterocyclene-, -heterocyclene-(CH2)mO-, -fused bicyclicene-, fused bicylicene - (CH2)mO-, -NR3-, -NOR3-, -CONH-, or -NHCO-; X and Y are independently C1-C4 alkylene, substituted alkylene, or together X, Y, and W combine to form -(CH2)n-AA-; R1 is independently halo, C1-C4 alkyl, hydroxy, C1-C4 alkoxy, haloalkyl, nitro, NR2R5, or -NHCO (C1-C4 alkyl); R3 is hydrogen, (CH2)maryl, C1-C4 alkyl, -COO (C1-C4 alkyl), -CONR4R5, - (C=NH)NH2, -SO(C1-C4 alkyl), -SO2(NR4R5), or -SO2(C1-C4 alkyl); R4 and R5 are independently hydrogen, C1-C4 alkyl, phenyl, benzyl, or combine with the nitrogen to which they are bonded to form a saturated or unsaturated 5 or 6 member ring; AA is an amino acid residue; m is independently 0, 1, 2, or 3; and n is independently 2, 3, 4, or 5; or a pharmaceuticaly acceptable salt or solvate thereof. 212 ו' בניסן התשס" ד - March 28, 2004

Description

Ref: 4439/97 122092/4 C WV ? ΓΝΰ1Ί3 ",2D13T]7 Ώ"Γ2 1_1Π1Π DFI UJ CYCLIC 3,4-BIS(3-INDOLYL)MALEIC ANHYDRIDE AND 3,4-BIS(3- INDOLYL)MALEIMIDE INTERMEDIATES FOR PROTEIN KINASE C INHIBITORS X-8951A -1- This application is a division from application no. 111850.

Background of the Invention Protein kinase C (PKC) consists of a family of closely related enzymes that function as serine/threonine kinases.

Protein kinase C plays an important role in cell-cell signaling, gene expression, and in the control of cell differentiation and growth. At present, there are currently at least ten known isozymes of PKC that differ in their tissue distribution, enzymatic specificity, and regulation. Nishizuka Y. Annu . Re . Biochem. 58: 31-44 (1989); Nishizuka Y. Science 258: 607-614 (1992) .

Protein kinase C isozymes are single polypeptide chains ranging from 592 to 737 amino acids in length. The isozymes contain a regulatory domain and a catalytic domain connected by a linker peptide. The regulatory and catalytic domains can be further subdivided into constant and variable regions. The catalytic domain of protein kinase C is very similar to that seen in other protein kinases while the regulatory domain is unique to the PKC isozymes. The PKC isozymes demonstrate between 40-80% homology at the amino acid level among the group. However, the homology of a single isozyme between different species is generally greater than 97%.

Protein kinase C is a membrane-associated enzyme that is allosterically regulated by a number of factors, including membrane phospholipids, calcium, and certain membrane lipids such as diacylglycerols that are liberated in response to the activities of phospholipases . Bell, R.M. and Burns, D.J., J. Biol. Chem. 266: 4661-4664 (1991); Nishizuka, Y. Science 258: 607-614 (1992) . The protein kinase C isozymes, alpha, beta-1, beta-2 and gamma, . require membrane phospholipid, calcium and diacylglycerol/phorbol esters for full activation. The delta, epsilon, eta, and theta forms of PKC are calcium-independent in their mode of activation. The zeta and lambda forms uf PKC are X-8951A -2- independent of both calcium and diacylglycerol and are believed to require only membrane phospholipid for their activation.

Only one or two of the protein kinase C isozymes may be involved in a given disease state. For example, the elevated blood glucose levels found in diabetes lead to an isozyme-specific elevation of the beta-2 isozyme in vascular tissues. Inoguchi et al . , Proc. Natl. Acad. Sci. USA 89: 11059-11065 (1992) . A diabetes-linked elevation of the beta isozyme in human platelets has been correlated with their altered response to agonists. Bastyr III, E.J. and Lu, J. Diabetes 42 : (Suppl. 1) 97A (1993) . The human vitamin D receptor has been shown to be selectively phosphorylated by protein kinase C beta. This phosphorylation has been linked to alterations in the functioning of the receptor. Hsieh et al., Proc. Natl. Acad. Sci. USA 88 : 9315-9319 (1991); Hsieh et al . , J. Biol. Chem. 268: 15118-15126 (1993) . In addition, recent work has shown that the beta-2 isozyme is responsible for erythroleukemia cell proliferation while the alpha isozyme is involved in megakaryocyte differe&tiation in these same cells. Murray et al., J. Biol. Chem. 268: 15847-15853 (1993).

The ubiquitous nature of the protein kinase C isozymes and their important roles in physiology provide incentives to produce highly selective PKC inhibitors. Given the evidence demonstrating linkage of certain isozymes to disease states, it is reasonable to assume that inhibitory compounds that are selective to one or two protein kinase C isozymes relative to the other PKC isozymes and other protein kinases are superior therapeutic agents . Such compounds should demonstrate greater efficacy and lower toxicity by virtue of their specificity.

The microbial indolocarbazole, staurosporine, is a potent inhibitor of protein kinase C that interacts with the catalytic domain of the enzyme. Tamaoki et al., Biochem.

Biophys. Res. Commun. 135: 397-402 (1986); Gross et al . , Biochem. Pharmacol. 40: 343-350 (1990) . However, the therapeutic usefulness of this molecule and closely related X-8951A -3- compounds is limited by the lack of specificity for protein kinase C over other protein kinases. Ruegg, U.T. and Burgess, G.M., Trends Pharmacol. Sci. 10: 218-220 (1989). This lack of selectivity results in unacceptable toxicity in this class of molecules.

An additional class of compounds related to staurosporine, the bisindolemaleimides, has been the focus of recent work. Davis et al., FEBS Lett. 259: 61-63 (1989); Twoemy et al., Biochem. Biophys. Res. Commun. 171: 1087-1092 (1990); Toullec et al., J. Biol. Chem. 266: 15771-15781 (1991); Davis et al., J. Med. Chem. 35: 994-1001 (1992); Bit et al., J. Med.

Chem. 36 : 21-29 (1993) . Some of these compounds have demonstrated selectivity for protein kinase C over other protein kinases.

Although compounds that demonstrate specificity to protein kinase C have been discovered, very little is known regarding isozyme selectivity. For example, analysis of the isozyme selectivity of staurosporine, shows little isozyme selectivity with the exception of poor inhibition of the zeta isozyme relative to the other isozymes. McGlynn et al., Cell. Biochem. .4.9: 239-250 (1992); Ward, N.E., and O'Brian, C.A., Molec. Pharmacol. 41: 387-392 (1992). Studies of the PKC-selective compound, 3- [1- (3-dimethylaminopropyl) -indol-3-yl] -4-( lH-indol-3-yl) -lH-pyrrole-2, 5-dione, suggest a slight selectivity for the calcium dependent isozymes. Toullec et al., J. Biol. Chem. 266: 15771-15781 (1991). Subsequent studies of this compound observed no difference, or possibly slight selectivity, for alpha over beta-1 and beta-2 isozymes. Martiny-Baron et al . , J. Biol. Chem. 268: 9194-9197 (1993); Wilkinson, et al., Biochem. J. 294: 335-337 (1993). Therefore, despite years of research and the identification of classes of compounds that inhibit protein kinase C versus other protein kinases, there remains a need for therapeutically effective isozyme-selective inhibitors.

Application no. 111850 provides novel, potent protein kinase C inhibitors. The compounds of said invention are 122092/2 -4- selective to protein kinase C over other kinases and are, quite surprisingly, highly isozyme-selective . As selective inhibitors the compounds are useful in treating conditions associated with diabetes mellitus and its complications, ischemia, inflammation, central nervous system disorders, cardiovascular disease, dermatological disease and cancer.

Application 111850 provides compounds of Formula X: wherein: Q is a group -N (! )-; R2 is hydrogen, CH3C0-, H2, or hydroxy; W-is -0-, -S-, -SO-, -S02-, -CO-, C2-C6 alkylene, substituted alkylene, C2-C6 alkenylene, -arylene-, -arylene- (CH2)m0-/ -heterocyclene-, -heterocyclene- (CJ'2 ) m0~/ -fused bicyclicene-, -fused bicyclicene- (CH2 )mO-, -NR3-, -NOR3-, -C0NH-, or -NHC0-; X and Y are independently C1-C4 alkylene, substituted alkylene, or together X, Y, and combine to form - (CH2 ) n-AA-; Rl is independently halo, C1-C4 alkyl, hydroxy, C1-C4 alkoxy, haloalkyl, nitro, NR4R5, or -NHCO(Ci-C4 alkyl;; R3 is hydrogen, (CH2)maryl, C1-C4 alkyl, -C00(Ci-C4 alkyl), -CONR4R5, -(C=NH)NH2, -S0(Ci~C4 alkyl), -SO2 ( R4R5) , or -SO2 (C1-C4 alkyl) ; R4 and R5 are independently hydrogen, C1-C4 alkyl, phenyl, benzyl, or combine with the nitrogen to which they are bonded to form a saturated or unsaturated 5 or 6 member ring; AA is an amino acid residue; m is independently 0, 1, 2, or 3; and -5- 122092/2 n is independently 2, 3, 4, or 5.

The compounds of formula (X) in which Q is a group -N (! )-, referred to hereinafter as compounds of formula I: are protein kinase C inhibitors.

The compounds of formula X in which Q is a group V, referred to hereinafter as compounds of the formula II: are intermediates in the synthesis of compounds of formula I, and are part of the present invention.

An aspect of the present invention is a process for preparing the compounds of Formula II, which comprises: Combining a mixture of a compound at a concentration of about 1.5 molar to about 0.001 molar of the formula: wherein: .

V is 0 .or N-CH3; Rl is independently halo, C1-C4 alkyl, hydroxy, C1-C4 aikoxy, haioalkyl, nitro,- . NR4R5 , or -NKCO (C1-C4 alkyl); m is independently 0, 1, 2, or 3; and an alkylating agent at a concentration of about i .5 molar to about 0.001 molar of the formula: L-X-W-Y-L wherein L is a leaving group; ■W is -0-, -S-., -SO-, ,-£302-, -CO-, C2-C.6 alkylene, substituted alkylene, C2~Cg aikenylene, -aryiene-, -arylene- (CK2-) mO~ -heterocyclene-, -heterocyciene- (CK2)mO-, . -fused bicyclice e-, -fused bicyclicene- (CK2)mP~ -NR3-, -NOR3-, -C0NH-, or -NHCO-; X and Y are independently CT-C4 alkylene or substituted alkylene; R3 is hydrogen,. (CH2).maryl, C1-C4 alkyl, -CCO(Ci-C4 . alkyl), -CONR4R5, -(C=K)NH2, -SO (C1-C4 alkyl) , -SO2 (NR4R5) , or -SO2 (C1-C4 alkyl) ; R and R5 are independently hydrogen, C1.-C4 alkyl, 'phenyl, benzyl, or combine wit the nitrogen to which they. are. bonded to form a saturated or unsaturated 5 or 6 member ring; m is independently 0, Y, 2, or 3; with about 0.5 to about 10 equivalents ' of CS2CO3 at a rate from, about .0.1 mL/hour to about 2.0 mL/hour in a polar aprotic solvent.

Yet another process of preparing the compounds of Formula II, comprises combining a compound of the formula: 122092/4 wherein: 1.2 i≤ a leaving group; V is -0- or N-CH3; W is -0-, -S-, -SO-, -302-, -CO-, C2-CS alkyiene, substituted alkyiene, C2-C5 alkenylene, -arylene-, -arylene- (CH2) VBP~, -heterccyclene-, -heterocyciene- (CK2)ELO-, -fused bicyclicene-, -fused bicyciicene- (CK2 ) -NR3--NOR3-, -C0NH-, or -NHC0-; X and Y are independently C1-C4 alkyiene or substituted alkyiene; Rl is independently halo, C1-C4 alkyi, hydroxy, C1-C4 alkoxy, haloalkyl, nitro, NR4R5, or -NHC0(Ci~C4 alkyi); R3 is hydrogen, (CK2)rriaryl, C1-C4 alkyi, -000( -04 alkyi), -CONR4R5, -(C=NH)MH2, -SQ(C1_-C4 alkyi), -S02(NRaR5), or -S02( i-C aikyl) ; R4 and R5 are independently hydrogen, C1-C4 alkyi, phenyl/ benzyl, or combine with the nitrogen to which they are bonded to form a saturated or unsaturated 5 or 6 member ring; in is independently 0, 1, 2, or 3; at a concentration of about 3 molar to about 0.001 molar with about 0.5 to about 10 equivalents of C52CO3 at a rate from about 0.1 mL/hour to about 2.0 mL/hour in a polar aprotic solvent.

A further aspect of the invention of application no. 111850 are compounds of formula I for use as medicaments for inhibiting Protein Kinase C, for example by administering to a mammal in need of such treatment a pharmaceutically effective amount of a compound of the Formula I. Also included in application 111850 are compounds of formula I for use as medicaments for selectively inhibiting the beta-1 and beta-2 protein kinase C isozymes, for example by administering to a mammal in need of such treatment a pharmaceutically effective amount of a compound of the Formula I.

Application no. 111850 further provides compounds of formula I for use as medicaments for treating conditions that protein kinase C has demonstrated a role in the pathology, such as ischemia, inflammation, central nervous system disorders, cardiovascular disease, dermatological disease, and cancer, for example by administering to a mammal in need of treatment a pharmaceutically effective amount of a compound of the Formula I.

Compounds of formula I are particularly useful in treating diabetic complications. Therefore, application 111850 further provides compounds of formula I for use as medicaments for treating diabetes mellitus, for example by administering to a mammal in need of such treatment a pharmaceutically effective amount of a compound of the Formula I.

A final aspect of the invention of application no. 111850 are pharmaceutical formulations comprising a compound of Formula I together with one or more pharmaceutically acceptable excipients, carriers, or diluents.

As noted above, the application no. 111850 provides compounds of the Formula I which selectively inhibit protein kinase C. The preferred compounds of the invention of application no. 111850 are those of Formula I wherein the moieties -X-W-Y-contain 4 to 8 atoms, which may be substituted or unsubstituted. Most preferably, the moieties -X-W-Y- contain 6 atoms.

Other preferred compounds are those compounds of Formula I wherein m is 0 and R2 is hydrogen; and W is a substituted alkylene, -0-, -S-, -C0NH-, -NHCO- or -NR3-. Particularly preferred compounds are compounds of the Formula la: X-8951A -9- wherein Z is -(CH2)p- or - (CH2 ) p-0- (CH2 ) p-; R6 is hydroxy, -SH, C1-C4 alkyl, (CH2)maryl, -NH(aryl), or -NR4R5; R4 is hydrogen or C1-C4 alkyl; R5 is hydrogen, C1-C4 alkyl, or benzyl; p is 0, 1, or 2; and ra is independently 2 or 3. Most preferred compounds of the Formula la are those wherein Z is CH2; R6 is -NH2 or N(CH3)2- Other preferred compounds are compounds wherein W is -0-, Y is substituted alkylene, and X is alkylene. These compounds are represented by Formula lb: wherein Z is -(CH2)p-; 6 is NR4R5; R4 and R5 are independently H or C1-C4 alkyl; p is 0, 1, or 2; and m is independently 2 or 3. Most preferred compounds of the Formula lb are those wherein p is 1; and R4 and R5 are methyl.

The term "halo" represents fluorine, chlorine,, bromine, or iodine.

The term "C1-C4 alkyl" represents a cyclo, straight or branched chain alkyl group having from one to four carbon atoms such as methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n- - - butyl, isobutyi, sac-butyl, t-butyl and tha lika. A haioaikyi is one' such aikyl substituted with one or mora halo atoms, preferably one to three halo atoms. An example of■ haioaikyi is trifluoromethyl . A CT.-C alkoxy is a C1.-C4 aikyl group covaientiy bondad by an -0- linkage.

The term "C1-C4 aikylene" represents a one to four carbon, straight aikylene moiety of the formula -(CK2)r-wherein r is one to four. Examples of C1-C4 aikyier.e inciuda methylene, ethylene, trimethyiane, methylethyiene, tetramethyiene, and the like. Similarly, a "C2-C6 aikylene" represents a two to six carbon, straight aikylene moiety.

Preferably, C2-C6 aikylene is a two to four carbon aikylene.

The term "C2-C g alkenyiene" represents a two to six carbon, straight or branched hydrocarbon containing one or more double bonds, preferably one or two doubia bonds.

Examples of a C2~C g alkenyiene include ethenyiene, propenyiene, 1 , 3 - butadienediyi, and 1 , 3 , 5 -hexatrier.ediyi .

The term "aryl" represents a substituted or unsubstituted phenyl or naphthyl. Aryl may be optionally substituted with one or two groups independently selected from hydroxy, carboxy,. C i~C4 alkoxy, -0 alkyi, haloalkyi, nitro, -NR4R5, -NHC0(Ci-C4 aikyl) , -NHCO (benzyl) , -NHCO (phenyl) , SH, S(Ci-C4 aikyl), -OCO(C].-C4 aikyl), -S02(NR4R5), -SO2 (C1-C4 aikyl) , -SO2 (phenyl) , or halo. The term (CH2)m yl is preferably benzyl or phenyl.

The above definition of the term "aryl" applies correspondingly to the term "arylene".

The term "substituted aikylene" represents a- moiety or the formula: wherein Z is- -(CH2)p- o - (CH2) p-Q- (CH2) p-; Rg is C1-C4 aikyl, C1-C4 alkoxy, (CH2)marY-' (CK?) aryioxy, hydrox , carboxy, -COO ( C i-C4 aikyl) , -COO ( (CK2 ) maryl) , -C0( Ci-Cd aikyl), -NR4 R5, -N ( ) (O 5).. -11- 122092/2 -NH(CH2)maryl, -NH (CH2)mPyridyl, -CONH ( (CH2)msryl) , -CONH(Ci-C4 alkyl), -NHCO(Ci-C4 alkyl), -NHCO(CH2)maryl, -OCONH(Ci-C4 alkyl), -OCONH (CH2) maryl, -NHCOO (alkyl) , -NHCOO (benzyl) , -NHS02(Ci-C4 alkyl), -NHSO2 (CH2) maryl, -CN, -SH, -S(Ci-C alkyl), -S(aryl), -SO2 (NR4R5) , -SO2 (C1-C4 alkyl), -SO(Ci-C4 alkyl), glycosyl, or heterocycle; R4 and R5 are independently hydrogen, C1-C4 alkyl, phenyl, benzyl, or combine with the nitrogen to which they are bonded to form a saturated or unsaturated 5 or 6 member ring; p is independently C, 1 or 2; and m is independently 0, 1, 2, or 3. Preferably Z is -CH2-; and 6 is C1-C4 alkyl, aryl, or -NR4R5.

The term "heterocycle" represents a stable, substituted or unsubstituted, saturated or unsaturated 5 or 6 membered ring, said ring having from one to four heteroatoms that are the same or different and that are selected rom the group consisting of sulfur, oxygen, and nitrogen; and when heterocycle contains two adjacent carbon atoms, the adjacent carbon atoms may be structured to form a group of the formula -CH=CH-; provided that (1) when the heterocyclic ring contains 5 members, the heteroatoms comprise not more than two 3 lfur or two oxygen atoms but not both; and (2) when the heterocyclic ring contains 6 members and is aromatic, sulfur and oxygen are not present. The heterocycle may be attached at any carbon or nitrogen which affords a stable structure. The heterocycle may be substituted with one or two groups independently selected from C1-C4 alkyl, C1-C4 alkoxy, hydroxy, oxo, acetyl, carboxy, haloalkyl, nitro, -NR4R5, -NHCO(Ci-C4 alkyl), -NHCO (benzyl) , -NHCO (phenyl) , SH, S(Ci-C4 alkyl), -OCO(Ci-C4 alkyl), -S02( R R5), -SO2 (C1-C4 alkyl), -SO2 (phenyl) , or halo. Examples of a heterocycle include pyrazole, pyrazolone, imidazole, acetylimidazole, isoxazole, triazole, tetrazole, oxazole, 1, 3-dioxolane, thiazole, oxadiazole, thiadiazole, pyridine, dipyridyl, pyrimidine, morpholine, pyrazine, - - pyrrolidine, piper dina, piperazine, oxazolidinone, imid zciidinor.e, and aminopyridir.e . e above 'definition of c e term "heterocycle" applies correspondingly to the term "hetarocyciena" .

- The term "giycosyi" represents a 5 or o carbon sugar, preferably selected from allasyi, altrasyl, glucosyi, mannosyi, g-ulasyl, idosyl, galactosyl, talosyi, arabinosyi, xylosyi, iyxcsyl, rhamnosyi, rihosyl, deoxy rancsyi , deoxy y anos l, and deoxyribosyi. The giycose may be azide substituted, O-acetylated, O-methylated, amino, mono, and di-aikylaiiiiac -substituted, . or acyiamino substituted.

The terra "fused bicyciic" represents a stable -fused bicyciic ring system of the formula: - wherein Ke'tero represents a substituted or unsubstituted, saturated or unsaturated 5 or 6 membered ring, said riag having', from .one to three heteroatoms that, are the same or different and that are selected from the group consisting of sulfur, oxygen, and nitrogen; and when Hetero contains two adjacent carbon, atoms, the adjacent carbo atoms may be structured" to form a group, of the formula—CH=CK-; provided that (1) when the Hetero ring contains 5 members, the heteroatoms comprise- not more than two sulfur or two oxygen atoms but not both;, and (2) when the. Hetero ring contains ' a members and is aromacic, sulfur and oxygen are not present. The fused bicyciic may be attached at any carbon or nitrogen atom which affords a srabie structure. The fused bicyciic may be substituted with one or two' croups independently selected from C1-C aikyi,. C1-C aikoxy, hydroxy, oxo, carboxy, haloaikyi, nitre, -iiR-iH-s, -NHC0(Ci~C4 aikyi) , -NHCC (benzyl) , -SHCO (phenyl) , SH, S (C1-C4 aikyi), -OCC(Ci-C4 aikyi), -30 (H R5), -SO2 (C1-C4 aikyi), -S02 (phenyl) , or halo. -13- 122092/2 Examples of a fused bicyclic include indole, imidazo(l,2-a) pyridine, benzotriazole, benzimidazole, benzotriazole, benzoxazole, benzoxathiazole, quinoline, isoquinoline, phthalazine, quinazoline, quinazolinone, quinoxaline, and aminoisoquinoline .

The above definition of the term "fused bicyclic" applies correspondingly to the term "fused bicyclicene".

The term "amino acid residue" refers to moiety of the formula or wherein R represents the variable side chain of an amino acid and R7 is hydrogen or hydroxy. The variable side chain of an amino acid represents the atom or group bonded to an a-carbon atom also having bonded thereto a carboxyl and an amino group. For example, the variable region of the naturally occurring amino acids are of the formulas: (Ala) (Val) (Leu) (He) CH3-S-CH2-CH2-, H- , HO-CH2- , H2N-CH2-CH2-CH2-CH2- , (Met) (Gly) (Ser) (Lys) X-8951A FOR - 14 - (Thr) (Cys) (Tyr) (Asn) H2 (Arg) (His) (Gin) (As ) i (Glu) In addition to the naturally occurring amino acids, the term amino acid residue includes positional isomers and variants. Examples of positional isomers and variants represented by amino acid residue include: 2-Aminoadipic acid (Aad) , 3-aminoadipic acid (bAad) , β-alanine (bAla) , 2-aminobutyric acid (Abu), 4-aminobutyric acid (4Abu) , 6-aminocaproic acid (Acp) , 2-aminoheptanoic acid (Ahe) , 2-aminoisobutyric acid (Aib) , 3-aminoisobutyric acid (bAib) , 2-aminopimelic acid (Apm) , 2,4-diaminobutyric acid (Dbu) , desmosine (Des) , 2 , 2'-diaminopimelic acid (Dpm) , 2, 3 -diaminopropionic acid (Dpr) , N-ethylglycine (EtGly) , N-ethylasparagine (EtAsnJ, hydroxylysine (Hyl) , allohydroxylysine (aHyl) , 3-hydroxyproline (3Hyp) , 4-hydroxyproline (4Hyp), isodesmosine (Ide) , allo-isoleucine (alle) , naphthy1glycine, N-methylglycine ( eGly) , N-methylisolGucine ( elle) , N-methyllysine (MeLys), norvaline (Nva) , norleucine (Nle) , ornithine (Orn) , phenylglycine, cyanoalanine (CA) , γ -carboxyglutamate, O-phosphoserine, a -15- 122092/2 -naphthylalanine ( A) , β-naphthylalanine (bNA) , S-gaiactosyl cysteine, glycinamide, N-formylmethionine, tyrosine-O-sulfate and the like. These amino acid residues may be in either the D or L configuration. Unless otherwise specified, a reference to an amino acid will refer to the L configuration.

The term- "leaving group" as used in the specification is understood by those skilled in the art. Generally, a leaving group is any group or atom that enhances the electrophilicity of the atom to which it is attached for displacement. Preferred leaving groups are triflate, mesylate, tosylate, imidate, chloride, bromide, and iodide. If the alkylating agent contains an amino acid residue (i.e., X, W, and Y combine to form - (CH2) n~AA*") -.the leaving group attached to the carbox is preferably pentaflourophenyl ester or para-nitrophenyl ester.

The term "carboxy protecting group" as used in the specification refers to one of the ester derivatives of the carboxylic acid group commonly employed to block or protect the carboxylic acid group while reactions are carried out on other functional groups on the compound. The species of carboxy-protecting group employed is not critical so Long as the derivatized carboxylic acid is stable to the condition of subsequent reaction (s) and can be removed at the appropriate point without disrupting the remainder of the molecule. T. .

Greene and P. Wuts, Protective Groups in Organic Synthesis, John Wiley and Sons, New York, N.Y., 1991, Chapter 5, provide a list of commonly employed protecting groups. See also E. Haslam, Protective Groups in Organic Chemistry, J.G.W. McOmi.i, Ed., Plenum Press, New York, N.Y., 1973. A related term in "protected carboxy," which refers to a carboxy group esterified with a carboxy-protecting group as defined.

The term "hydroxy protecting group" as used in the specification refers to one of the ether or ester derivatives of the hydroxy group commonly employed to block or protect the hydroxy group while reactions are carried out on ottvjr functional groups on the compound. The species of hydroxy protecting group employed is not critical so long as the derivatized hydroxy group is stable to the condition cf subsequent reaction (s) and can be removed at the appropriate point without disrupting the remainder of the molecule. T.W. -16- 122092/2 Greene and P. Wuts, Protective Groups in Organic Syncl.esis, John Wiley and Sons, New York, N.Y., 1991, provide a list of commonly employed protecting groups. Preferred hydroxy protecting groups are tert-butyldiphenylsilyloxy (TBDPS), tert-butyldimethylsilyloxy (TBDMS) , triphenylmethyl (trityl), methoxytrityl, or an alkyl or aryl ester. A related ~nrm is "protected hydroxy," which refers to a hydroxy group f.sterified or etherified with a hydroxy protecting group as defined.

The term "amino protecting group" as used in the specification refers to substituents of the amino group commonly employed to block or protect the amino functionality while reacting other functional groups on the compound. The species of amino-protecting group employed is not critical so long as the derivatized amino group is stable to the condition of subsequent reaction (s) and can be removed at the appropriate point without disrupting the remainder of the molecule. T. W. Greene and P. Wuts, Protective Groups in Organic Synthesis, Chapter 7, provide a list of commonly employed protecting groups. See also J. W. Barton, Protective Groups in Organic Chemistry, Chapter 2. Preferred amino-protecting groups are t-butoxycarbonyl, pthalimide, a cyclic alkyl, and benzyloxycarbonyl. The related term "protected amino" defines an amino group substituted with an amino protecting group as defined.

The term "-NH protective groups" as used ii the specification refers to sub-class. of amino protecting groups that are commonly employed to block or protect the -NH functionality while reacting other functional groups on the compound. The species of protecting group employed is not critical so long as the derivatized amino group is stable to the condition of subsequent reaction (s) and can be removed at the appropriate point without disrupting the remainder of the molecule. T. W. Greene and P. Wuts, Protective Groups in Organic Synthesis, Chapter 7, page 362-385, provide a list of commonly employed protecting groups. Preferred -NH projecting groups are carbamate, amide, alkyl or aryl sulfonamide. The related term "protected -NH" defines a group substi uted with an -NH protecting group as defined.

X-8951A FOR - 17 - The term "pharmaceutically effective amount", as used herein, represents an amount of a compound of the invention that is capable of inhibiting P C activity in mammals. The particular dose of the compound administered according to this invention will, of course, be determined by the particular circumstances surrounding the case, including the compound administered, the route of administration, the particular condition being treated, and similar considerations. The compounds can be administered by a variety of routes including the oral, rectal, transdermal, subcutaneous, topical, intravenous, intramuscular or intranasal routes. For all indications, a typical daily dose will contain from about 0.01 mg/kg to about 20 mg/kg of the active compound of this invention. Preferred daily doses will be about 0.05 to about 10 mg/kg, ideally about 0.1 to about 5 mg/kg. However, for topical administration a typical dosage is about 1 to about 500 g compound per cm2 of an affected tissue. Preferably, the applied amount of compound will range from about 30 to about 300 μg/cm2, more preferably, from about 50 to about 200 μg/cm2, and, most preferably, from about 60 to about 100 μg cm2.

The term "treating, " as used herein, describes the management and care of a patient for the purpose of combating the disease, condition, or disorder and includes the administration of a compound of present invention to prevent the onset of the symptoms or complications, alleviating the symptoms or complications, or eliminating the disease, condition, or disorder.

The term "isozyme selective" means the preferential inhibition of protein kinase C beta-1 or beta-2 isozyme over protein kinase C isozymes, alpha, gamma, delta, epsilon, zeta,* and eta. In general, the compounds demonstrate a minimum of a eight fold differential (preferably a ten fold differential) in the dosage required to inhibit PKC beta-1 or beta-2 isozyme and the dosage required for equal inhibition of the alpha protein kinase C isozyme as measured in the PKC assay. The compounds demonstrate this differential across the range of inhibition and are exemplified at the IC50, i.e., a 50% inhibition. Thus, isozyme-selective compounds inhibit the beta-1 and beta-2 -18- 122092/2 isozymes of protein kinase C at much lower concentrations with lower toxicity by virtue of their minimal inhibition of the other PKC isozymes.

By virtue of their acidic moieties, the compounds of Formula I include the pharmaceutically acceptable base addition salts thereof. Such salts include those derived fro:n inorganic bases such as ammonium and alkali and alkaline earth rnstal hydroxides, carbonates, bicarbonates, and the like, as well as salts derived from basic organic amines such as aliphatic and aromatic amines, aliphatic diamines, hydroxy alkamines, and the like. Such bases useful in preparing the salts of this invention thus include ammonium hydroxide, potassium carbonate, sodium bicarbonate, calcium hydroxide, methylamine, diethylamine, ethylenediamine, cyclohexylamine, ethanoiamine and the like.

Because of the basic moiety, the compounds of Formula I can also exist as pharmaceutically acceptable acid addition salts. Acids commonly employed to form such salts include inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, as well as organic acida such as para-toluenesulfonic, methanesulfonic, oxalic, para-bromophenylsulfonic, carbonic, succinic, citric, benzoic, acetic acid, and related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphat , mono-hydrogenphosphate, dihydrogenphosphate, etaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, 2-butyne-l,4 dioate, 3-hexyne-2, 5-dioate, benzoate, chlorobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, xylenesulfonate, phenylacetace, phenylpropionate, phenylbutyrate, citrate, lactate, hippurate, β-hydroxybutyrate, glycolate, maleate, tartarate, methanesulfonate, propanesulfonate, naphthalene-l-sulfor.ate, naphthalene-2-sulfonate, mandelate and the like salts.

In addition to pharmaceutically-acceptable salts, other salts are described herein.. They may serve as X-8951A FOR - 19 - intermediates in the purification of the compounds, in the preparation of other salts, or in the identification and characterization of the compounds or intermediates.

The pharmaceutically acceptable salts of compounds of Formula I can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, ethyl acetate and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent. Such solvates are within the, scope of the present invention.

It is recognized that various stereoisomeric forms of the compounds of Formula I may exist; for example, W may contain a chiral carbon atom in the substituted alkylene moiety. The compounds are normally prepared as racemates and can conveniently be used as such, but individual enantiomers can be isolated or synthesized by conventional techniques if so desired. Such racemates and individual enantiomers and mixtures thereof: form part of the present invention.

The invention also encompasses the pharmaceutically acceptable prodrugs of the compounds of Formula I. A prodrug is a drug which has been chemically modified and may be biologically inactive at its site of action, but which may be degraded or modified by one or more enzymatic or other in vivo processes to the parent bioactive form. This prodrug should have a different pharmacokinetic profile than the parent, . enabling easier absorption across the mucosal epithelium, better salt formation or solubility, and/or improved systemic stability (an increase in plasma half-life, for example) . Typically, such chemical modifications include the following: 1) ester or amide derivatives which may be cleaved by esterases or lipases; 2) peptides which may be recognized by specific or nonspecific proteases; or 3) derivatives that accumulate at a site of action through membrane selection of a prodrug form or a modified prodrug form; or any combination of 1 to 3, supra . Conventional procedures for the selection and preparation of suitable prodrug -20- 122092/2 derivatives are described, for example, in H, Bundgaard, Design of Prodrugs, (1985) .

The synthesis of certain bis-indole-N-maleimide derivatives is described in Davis et al. U.S. Patent 5,057,614, and in Israeli patent 89167, herein incorporated by reference. Generally, the compounds of the present invention may be prepared as follows: Rl, m, and halo are the same as previously defined. Halo is preferably chloro, bromo, or iodo. Compound III is preferably 2,3-dichloro N-methylmaleimide .

The reaction between Compound III and the indole, Compound IV, is commonly known as a Grignard reaction. The reaction is carried out in an inert organic solvent, such as toluene, at a temperature between room temperature and the reflux temperature of the reaction mixture. Most significantly, the reaction depicted in Scheme 1 is dependent on solvent conditions. When carried out in a Toluene : THF: ether solvent system, the reaction provides Compound V in greater than 80 percent yield and greater than 95 percent purity. The product is precipitated from the reaction mixture with ammonium chloride, NH4CI. The resulting intermediate, Compound V, may be isolated by standard techniques. 3, 4-Bis (3-indolyl) -l-methyl-lH-pyrrole-2, 5-dione, Compound V, may then be converted by alkaline hydrolysis to the corresponding anhydride of the Formula VI by techniques known in the art and described in Brenner et al., Tetrahedron 44 : 2887-2892 (1988). Preferably, Compound V is reacted with 5 N o KOH in ethanol at a temperature ranging from 25 C to reflux.

X-8951A FOR - 21 - Compounds of the Formula V are generally more stable than the compounds of the Formula VI. Therefore, it is preferred that Compounds V are reacted in accordance with Scheme 2 to produce the compounds of Formula I. However, one skilled in the art would recognize that the compounds of the. Formula VI, may also be reacted according to Scheme 2.

Scheme 2 (VII) X, Y, and W are the same as previously defined. L is a good leaving group such as chloro, bromo, iodo, mesyl, tosyl, and the like. L may also be a hydroxy or other precursor that may be readily converted to a good leaving group by techniques known in the art. For example, the hydroxy may be readily converted to a sulfonic ester such as mesyl by reacting the hydroxy with methanesulfonyl chloride to produce the mesylate leaving group.

The reaction represented by Scheme 2 is accomplished by any of the known methods of preparing N-substituted indoles. This reaction usually involves approximately equimolar amounts of the two reagents, although other ratios, especially those wherein the alkylating reagent is in excess, are operative. The reaction is best carried out in a polar aprotic solvent employing an alkali metal salt or other such alkylation conditions as are appreciated in the art. When the leaving group is bromo or chloro, a catalytic amount of iodide salt, such as potassium iodide may be added to speed the reaction.

X-8951A FOR - 22 - Reaction conditions include the following: Potassium hexamethyldisilazide in dimethylformamide or tetrahydrofuran, sodium hydride in dimethylformamide.

Preferably, the reaction is carried out under slow reverse addition with cesium carbonate in either acetonitrile, dimethylformamide (DMF) , or tetrahydrofuran (THF) . The temperature of the reaction is preferably from about ambient temperature to about the reflux temperature of the reaction mixture .

One skilled in the art would recognize that the reaction described in Scheme 2 may be employed with compounds of the Formula Vila: L—Y' L — ' Vila X' and Y' are a protected carboxy, protected hydroxy, or a protected amine. After the alkylation of Scheme 2, X' and Y' may be converted to moieties capable of coupling to form w.

This method is the preferred method of preparing the compounds of Formula I wherein fa is -S-, -0-, or NR3. The coupling of X' and Y' to form the various ether, thioether or aminoether derivatives is known in the art and described in, for example, Ito, et al., Chem. Pharm. Bull. 41(6): 1066-1073 (1993); Kato, et al., J. Chem. Pharm. Bull. 34: 486 (1986); Goodrow, et al. Synthesis 1981: 457; Harpp, et al., J. Am. Chem. Soc. 93: 2437 (1971); and Evans, et al., J. Org. Chem. 50: 1830 (1985).

One skilled in the art would also recognize that the compounds of Formula V may be converted to the compounds of Formula II in a two step synthesis as described in Scheme 3.

X-8951A -23- Scheme 3 Rl, X, , Y, V and L are the same as previously defined. L2 is a protected hydroxy or other group that may be readily converted to a good leaving group by techniques known in the art. The coupling between Compound V or VI and Compound VIII is an alkylation as previously discussed. The monoalkyl-ated intermediate, IX, is deprotected, and L2 is converted to a leaving group. For example, if the hydroxy is protected with t-butyldimethylsilyl (TBDMS) , TBDMS is selectively removed using acidic methanol. The resulting free hydroxy is then converted to a leaving group, such as an alkyl halide, preferably an alkyl iodide or bromide (CBr4 in triphenylphosphine) or sulfonate (mesyl chloride in triethylamine) . The macrolide is then formed by alkylating under slow reverse addition to a solution of base, such as potassium hexamethyldisilazide, or sodium hydride but preferably CS2CO3 in a polar aprotic solvent such as acetoni-trile, DMF, THF at temperatures ranging from ambient to reflux.

Schemes 2 and 3 exemplify the process of the present application. Most unexpectedly, the compounds of the Formula II may be prepared in substantially higher yield when the alkylation is carried out under slow reverse addition to CS2CO3 in a polar aprotic solvent. Slow reverse addition involves ¼ combining a mixture of compound and alkylating agent (Scheme 2) or the compound (Scheme 3) with the base at a rate from about 0.1 mL/hour to about 2.0 mL/hour. The concentration of each reagent in the mixture is about 1.5 molar to about 0.001 molar. When carried out with the monoalkylated compound (Scheme 3) the concentration is from about 3 molar to about 0.001 molar. The slow addition results in a concentration of reagents in the reaction vessel of about 0.01 umolar to 1.5 molar. One skilled in the art would recognize that at a higher rate of addition a lower concentration of reagents could be used in the reaction.

X-8951A -24- Likewise, at a slower rate of addition, a higher concentration of reagents could be used in the reaction. Preferably, the compound is added at about .14 mL/hour with the compound and the alkylating agent at 0.37 molar. It is preferred that the CS2CO3 be added in excess -- most preferably a 4:1 ratio CS2CO3 to alkylating agent. Preferred polar aprotic solvents are acetonitrile, dimethylformamide (DMF) , acetone, dimethylsulfoxide (DMSO) , dioxane, diethylene glycol methyl ether (diglyme) , tetrahydrofuran (THF) , or other polar aprotic solvents in which the reagents are soluble. The reaction is carried out at temperatures ranging from about 0°C to reflux. One skilled in the art would recognize that the ratio of the mixture of the compound and alkylating agent is not critical. However, it is preferred that the reagents are mixed in a ratio of 0.5 to 3 equivalents of each other. Most preferably, the reagents are mixed 1:1.

When V is N-CH3, Compound II is converted to the corresponding anhydride (V is 0) by alkaline hydrolysis.

Alkaline hydrolysis involves reacting the compound with a base, such as sodium hydroxide or potassium hydroxide, in C1-C4 alcohol (preferably ethanol) , DMSO/water, dioxane/water, or acetonitrile/water a^ a temperature ranging from about 25°C to preferably about reflux. The concentration of the reactants is not critical.

The anhydride (V is 0) is converted to the maleimide of Formula I by ammonolysis. Ammonolysis involves reacting the anhydrtide with an excess of hexamethyldisilazane or an ammonium salt (ammonium acetate, bromide, or chloride) and C1-C4 alcohol (preferably methanol) in an polar aprotic solvent such as DMF at room temperature. Preferably, the hexamethyldisilazane or an ammonium salt is reacted at a ratio greater than about 5:1 equivalents of anhydride.

Yet another method of preparing the compounds of Formula I is outlined in Scheme 4. This method is particularly useful when W is -NH and X or Y is a substituted alkylene. 122092/2 X-8951A FOR - 25 (XII) (XIII) (XIV) (XV) Ac is acetyl. Ri, R6» Z, n, and m are the same as previously defined. The alkylation of Compound VI with X occurs under conditions previously described and known in the art.

Likewise, alkylation of Compound XI with the cc-halo ketone, Compound XII, occurs under conditions previously discussed. The conversion of the anhydride to the maleimide, Compound XV, occurs as previously described. For example, the anhydride may be converted to the bis-indole maleimide by reacting the anhydride with hexaraethyldisilazane and methanol in an inert organic solvent such as DMF at room temperature.

X-8951A FOR - 26 - The protected hydroxy, represented by OAc, is readily hydrolyzed to form an alcohol (for example, K2CO3 in aqueous methanol and THF) . The resulting alcohol is converted to a leaving group by methods appreciated in the art such as reacting the alcohol with mesyl chloride in triethylamine at 0°C. The leaving group is substituted with an azide, such as NaN3 in DMF at 50°C. The resulting azide is reduced to form the amine by employing Lindlar's catalyst in the presence of ¾. The macrocycle is allowed to close via an intramolecular Schiff base. The Schiff base is reduced under standard conditions, such as NaCNBH3 or other reducing agents, to form the macrocycles of Formula I.

Yet another method of preparing the compounds of Formula I is outlined in Scheme 5. This method is particularly useful when X, W, and Y are taken together to form -(CH2)n_AA_- S heme 5 (XIX) (XVIII) Rl, Ac, V, m, and n are the same as previously defined. Ρχ represents an amino protecting group. R represents the variable X-8951A FOR - 27 - side chain of an amino acid. The acylation of Compound XVI with an activated amino acid (such as the para-nitrophenyl ester, illustrated) is carried out using 18-crown-6 and KF in THF, DMF, or dimethoxyethane at room temperature as described in Klausner, et al., J. Chem. Soc. PERKIN I 607-631 (1977); and Nakagawa, et al., J. Am. Chem. Soc. 105: 3709-3710 (1983). Closure of the macrocycle to form Compound XIX is carried out via formation of the intramolecular Schiff base as described in Scheme 4.

An additional method of preparing the compounds of Formula I and a preferred method when W is -CO H- or -NHCO-, is described in Scheme 6.

Scheme 6 (XXIII) ( xii) Rl, Ac, V, Pi, m, and n are the same as previously defined. The reaction between Compound XX and Compound XXI occurs in the presence of ethyl diisopropylamine in methylene chloride at 0°C. The macrocycle is formed via an intramolecular alkylation of the X-8951A FOR - 28 - free indole nitrogen and the oc-halo carbonyl terminus under alkylating conditions previously described. The protected maleimide is deprotected as previously discussed to produce the Compound XXIII.

An alternative method of preparing the intermediates, Compounds XI and XX is described in Scheme 7.

Scheme 7 Ac is the same as previously defined; P is an indole protecting group such as t-butoxycarbonyl or other indole protecting group known in the art. T. W. Greene and P. Wuts, Protecting Groups in Organic Synthesis. Chapter 7, page 385. The reaction described in Scheme 7 is known as a Perkin Condensation. The reaction is described in Hill et al., J. Med. Chem. 36 : 21-29 (1993). Generally, oxalyl chloride is added at between -78°C and the reflux temperature of the mixture (prefferably at 0°C) to an anhydrous solution of Compound XXIV in inert organic solvent such as a halogenated aliphatic hydrocarbon like methylene chloride. After about one to three hours, the volatiles are removed. The resulting solids are dissolved in a dry halogenated aliphatic hydrocarbon solvent, e.g. methylene chloride; and added to Compound XXV in the presence of an acid binding agent, preferably a tertiary amine such as triethylamine, at. room temperature.

The resulting anhydride, Compound XI is reacted in accordance with Schemes 4 or 5 or converted to the maleimide or a protected maleimide as previously discussed. 122092/2 X-8951A FOR - 29 - The protected hydroxy (preferably OAc, illustrated) of Compound XI may be converted to an alcohol by techniques known in the art. For example, Compound XI is reacted with NH4OH or aqueous ammonia in DMF at elevated temperatures, e.g. 140°C.

The resulting alcohol is converted to the amine. Compound XX, by methods known in the art. For example, the alcohol in dichloromethane and collidine under a nitrogen atmosphere is reacted with triflic anhydride in dichloromethane . After approximately two hours, the mixture is treated with aqueous ammonia. The resulting amine, Compound XX is then reacted in accordance with Scheme 6.

An intermediate of the present invention is prepared in accordance with Scheme 8. This scheme is particularly useful in preparing compounds wherein W is -0-, Y is substituted alkylene, and X is alkylene. lAAAJ (XXIX) (XXVIII) R8 is N3, NH-protecting group, amine protecting group, or hydroxy protecting group; m is independently 0, 1, 2, or 3; and L is a good leaving group such as chloro, bromo, iodo, mesyl, tosyl and the like. L is preferably mesyl. R8 is preferably a protected hydroxy, most preferably -Otrityl.

X-8951A FOR - 30 - Scheme 8 presents a stereoselective synthesis of the linker portion (-X-W-Y-) of the macrocycle. The S-enantiomer is • illustrated above; however, one skilled in the art would recognize that the complimentary- enantiomer or mixture of enantiomers could be prepared in an analogous manner.

Furthermore, one skilled in the art would recognize that an analogous reaction with a methyl substituted epoxide or Grignard reagent could be used to prepare the various linkers (-X-W-Y-) containing a methyl substituted alkylene.

In the above reaction, the epoxide, Compound (XXVI) , is opened using a Grignard reagent. The reaction is carried out in the presence of copper complexing agent; however other alkylating conditions are operative. The reaction is carried out in an inert solvent at a temperature between -30°C and reflux temperature of the reaction mixture. The reaction produces Compound (XXVII) which may be further reacted without purification. Compound (XXVII) is allylated under general conditions known in the art for preparing ethers. The reaction illustrated ■ in Scheme 8 is a Williamson synthesis. The formation of sodium alkoxide using NaH, NaOH, or KOH followed by allylation with allyl bromide produces the diene, Compound (XXVIII) . Compound (XXVIII) is converted to the alcohol, Compound (XXIX), under standard techniques. For example, Compound (XXVIII) can be converted to an ozonide by treating with ozone at low temperatures. The ozonide is then reduced with NaBH4, LiAlH4, BH3 or catalytic hydrogenation with excess H2 to produce the alcohol, Compound (XXIX) . The hydroxy moieties of Compound (XXIX) are converted to leaving group, L, by standard techniques such as reacting the alcohol with mesyl chloride in triethylamine.

In all of the above schemes, it is preferred that the reactions be carried out with appropriate protecting groups. In particular, it is preferred that Ri is protected during the alkylations and/or acylations and subsequently deprotected.

Likewise, if R6 is to be a - R4R5, the reactions are best carried out with an amino protecting group. However, one skilled in the art recognizes that many of these reactions can be performed without protecting groups if the appropriate X-8951A -31- reaction conditions, blocking reagents, or the like are used. It is preferred that when W contains a hydroxy moiety, it is protected as tert-butyldiphenylsilyloxy (TBDPS) or triphenylmethyl (trityl) during the alkylation or acylation of the indole. The resulting compounds of Formula I may be isolated and purified by standard techniques.

Compounds III, IV, V, VII, Vila, VIII, X, XII, XVII, XXI, XXIV, XXV, XXVI and any other reagents required for the above reactions, are either commercially available, known in the art, or can be prepared by methods known in the art. For example, Compound III may be prepared by techniques described in Edge et al . , Chem. and Ind. 130 (1991); Compound IV is preferably prepared in situ by reacting an appropriately substituted indole with an alkylmagnesium halide such as ethylmagnesium bromide in a known manner.

The following examples and preparations are provided merely to further illustrate the invention. The scope of the invention is not construed as merely consisting of the following examples. Passages of the description which are not within the scope of the claims do^ not constitute a part of the invention. To aid one skilled in the art, the following structure is · provided to illustrate with a representative compound the nomenclature adopted herein: In the following examples and preparations, melting point, nuclear magnetic resonance spectra, mass spectra, high pressure liquid chromatography over silica gel, N, N-dimethylformamide, palladium on charcoal, tetrahydrofuran, and ethyl acetate are abbreviated M.Pt., N R, MS, HPLC, DMF, Pd/C, THF, and EtOAc X-8951A -32- 122092/2 respectively. The terms "NMR" and "MS" indicate that the spectrum was consistent with the desired structure.

Preparation 1 3, -bis- (3'-indolyl) -furan-2, 5-dione Sodium ethoxide (3.56 g, 50 mmol) was added to a solution containing 3, -dichloromaleic anhydride (5.56 g, 33.3 mmol) and methylamine hydrochloride (3.50g, 55.0 irimol) in 40 mL of acetic acid. The mixture was stirred under a CaCl2 drying tube at 25°C for 16 hours and then refluxed for 4 hours. The cooled mixture was poured into water (350 mL) and extracted with EtOAc (3 x 75 mL) . The combined organic extracts were washed with 100 mL portions of saturated aqueous NaHC03, water and brine and dried (MgSO-j) . The solvent was evaporated under reduced pressure. The residue was recrystallized from ethanol to give 3.82 g (64%) of 3, 4-dichloro-N-methyluialeimide as white crystals. Concentration of the mother liquor and chroraatography of the residue by radial preparative layer chromatography (Chromatotron, Harrison Research), gave an additional 0.81 g of 3, 4-dichloro-N-itiethylmaleimide, raising the yield to 77%.

A solution of indole (10.5 g, 90 mmol) in 175 mL of dry toluene was treated dropwise over 1 hour under N2 with a solution of ethylmagnesium bromide (1.0M in THF, 90 mL, 90 mmol) . After the addition was complete, the light-green solution was heated at 40°C for 30 minutes and then cooled to 25°C. A solution of 3, 4-dichloro-N-methylmaleimide (3.8 g, 21 mmol) in 50 mL of toluene was added over a 30-minute period.

The reaction mixture was heated at 100°C for 3 hours, then cooled to 25 °C, and quenched with 100 mL of 20 percent aqueous citric acid. The layers were separated. The aqueous phase was extracted with EtOAc (50 mL) . The combined organic layers were dried over anhydrous MgS04. The solvent was evaporated under reduced pressure. The residue was taken up in 30 mL of acetone and allowed to stand at 5°C for 40 hours. The solids were collected and washed with ice-cold ether to give 5.25 g (73 percent) of 3, -bis- (3 ' -indolyl) -l-methyl-lH-pyrrole-2, 5-dione s ared solid, M.Pt. 276-278°C. 122092/2 X-8951A -33- To a solution of 3, 4-bis- (3 ' -indolyl) -1-methyl-pyrrole-2, 5-dione in 150 mL of ethanol was added 5N KOH (50 mL) . The mixture was stirred 4 hours at 25°C and diluted with 150 mL of water. Most of the ethanol was evaporated under reduced pressure. The mixture was then acidified to pH 1. The precipitated product was filtered and washed with water. The crude product was dissolved in a minimum of CH2CI2 and slowly filtered through a two-inch column of silica gel eluting with 50 percent EtOAc in hexane to give the titled compound (3.10 g 79 percent) as a red solid. M. Pt. 225-228°C.

Preparation 2 2, 6-Bis (bromomethyl) pyridine To a mixture containing 2, 6-pyridinedimethanol (735 mg, 5.28 mmol) and triphenylphosphine (3.20 g, 12.2 mmol) in 35 mL of dry CH2CI2 at 0°C under 2 was added N-bromosuccinimide (2.16 g, 12.2 mmol) in portions over 10 minutes. The mixture was stirred 1 hour at 0°C and then allowed to stand at 5°C for 16 hours. Most of the solvent was removed under reduced pressure. Ether (100 mL) was added to the residue. The ether layer was decanted and concentrated to 20 mL then diluted with 3:1 hexane/EtOAc (50 mL) . The cloudy solution was placed in the refrigerator overnight. After evaporation of the solvents in vacuo, the crude product was recrystallized from hexane to afford 766 mg (55 percent) of 2,6-bis (bromomethyl) pyridine as a white crystalline solid. MS.

Preparation 3 (±) -3- (Benzyloxymethyl) -1, 6-dibromohexane A solution of potassium t-butoxide (1.0 M in THF, 8.27 mL, 8.27 mmol) was added dropwise to a solution of (±)-3 cyclohexene-l-methanol (853 mg, 7.60 mmol) in THF (35 mL) at 25°C under N2- The resultant mixture was stirred at 25°C for 30 minutes. Benzyl bromide (1.0 mL, 8.37 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours and then treated with saturated aqueous NH4CI (5 mL) and concentrated. The residue was dissolved in ether (50 mL) , X-8951A - 4- washed with water (20 mL) and brine (20 itiL) , and dried over MgS04. The solvent was evaporated under reduced pressure. The residue was subjected to radial chromatography on silica gel eluting with 5 percent EtOAc in hexane to give (±) -3- (benzyloxymethyl) -1-cyclohexene (1.42 g, 92 percent) as a colorless oil. NMR Ozone was bubbled through a solution of (±)-3- (benzyloxymethyl) -1-cyclohexene (1.35 g, 6.70 mmol) in CH2CI2 (65 mL) at -78 °C until the blue color of unreacted ozone persisted. The reaction mixture was allowed to warm to room temperature, while dry nitrogen was bubbled through the reaction. Borane-dimethyl sulfide complex (10.0 M in THF, 2.7 mL, 27.8 mmol) was added via syringe over several minutes, and the reaction mixture was allowed to stand at room temperature for 24 hours. The reaction mixture was treated with 5 percent aqueous HC1 (1 mL) and stirred vigorously for one hour. Solid NaHC03 was added until the mixture tested basic to litmus paper. The mixture was dried over anhydrous MgS0 . The reaction mixture was filtered and concentrated to afford the crude (±)-3- (benzyloxymethyl) -1, 1.49 g, ca. 100 percent) as an oil. This material, essentially a single spot on TLC analysis, Rf = 0.25, 25 percent EtOAc in hexane, was used directly in the next step without further purification.

N-Bromosu.ccinimide (2.49 g, 14.0 mmol) was added to a stirred mixture of (±) -3- (benzyloxymethyl) -1, 6 hexanediol (1.45 g, 6.10 mmol) and triphenylphosphine (3.67 g, 14.0 mmol) in dry CH2CI2 (50 mL) at 0°C under N2. After 12 hours, the reaction was concentrated and ether (100 mL) was added to the residue. The mixture was stirred 15 minutes; and the ether layer was decanted from the solids. This was repeated with 50 mL of ether. The combined ether extracts were concentrated to 50 mL then diluted with hexane (100 mL) . After standing at 5°C ■ overnight, the solution was decanted from the precipitated solids and concentrated to afford dibromide (±)-3-(benzyloxymethyl) -1, 6-dibromohexane (1.09 g, 49 percent) as a light yellow oil which was essentially homogeneous by TLC, Rf = 0.75 (10 percent EtOAc in hexane) . NMR X-8951A -35- 122092/2 Preparation 4 1- ( tsrt-butyldimethylsilyloxy) -4- ( tert-butyldiphenylsilyloxy) - butan-3-ol To an anhydrous CH2CI2 (110 m ) solution of 3-buten-l-ol (15 g, 0.21 mol) was added imidazole (28.6 g, 0.42 mol, 2 eq) , followed by tert-butyldimethylsilyl chloride (32 g, 0.22 mol) . After 90 minutes, the reaction was complete as indicated by TLC (10% EtOAc/hexane) . The CH2CI2 solution was transferred to a separatory funnel, diluted with CH2CI2 (110 mL) , washed with water (200 mL) , and brine (200 mL) . The organic layer was collected, dried over MgSO The above oil was dissolved in anhydrous CH2CI2 (250 mL) . Imidazole (30 g, 0.44 mol, 2.5 eq) was added to the solution as a solid with stirring. The resulting solution was cooled to 0°C. After cooling 15 minutes, a CH2CI2 (50 mL) solution of tert-butyldiphenylsilyl chloride (50 g, 0.18 mol, 1 eq) was added dropwise over 45 minutes. After the addition was complete, scirring was continued at 0°C for 2.5 hours. The solution was transferred to a separatory funnel, diluted with CH2CJ (250 ml.) , washed with water, brine, dried over MgSO-j, and filtered. The solvent was removed under reduced pressure to give X-8951A -36- the crude product as an oil. The crude product was purified by eluting (10% EtOAc/hexane) it through a short column of silica gel. The eluting solvent was removed in vacuo to leave a viscous oil of the titled intermediate. (78.1 g, 93 % overall yield) . MS Preparation 5 1- ( tert-butyldimethylsilyloxy) -3- (3-iodopropyloxy) -4- ( tert- butyldiphenylsilyloxy) -butane To a methylene chloride (20 ml) /cyclohexane (100 ml) solution of the alcohol of Preparation 4 was added allyl trichloroacetimidate (17.82 g, 88 mmols, 2.2 eq) under an N2 balloon followed by trifluoromethanesulfonic acid (50 μ /g of starting material, 0.92 ml). After 20 hours, the solution was filtered, and the filtrate was washed with saturated aqueous NaHC03, water, and then brine. The organic layer was collected and dried over MgS04. The solvent was removed to give an oil, which was purified by flash chromatography on silica' gel eluting with hexanes and increasing the polarity of the mobile phase to 5% ethyl acetate in hexanes over several liters to yield 19.27 g of the allylic ether, 1- ( tert-butyldimethylsilyloxy) -3- (2-propenyloxy) -4- ( tert-butyldiphenylsilyloxy) -butane as a light brown oil (97% yield). MS.

To a THF -(60 ml) solution of the above allyl ether (14.16 g, 28.38 mmols, 1 eq) was added 9-BBN (9-borabicyclo [3.3.1] nonane, 0.5 M solution in THF, 60 ml, 30 mmols, 1.1 eq) dropwise under nitrogen. After 3 hours, TIC (10% EtOAc in hexanes) of the reaction showed that the starting material had been consumed. To this solution was added 3M aqueous NaOH (10.41 ml, 31.22 mmols, 1.1 eq) followed by slow (1.5 hr) dropwise addition of 30% hydrogen peroxide (10.3 ml, 90.82 mmols, 3.2 eq) . The reaction temperature during the peroxide quench was kept below 50°C(ice bath).

After 30 minutes, sodium chloride was added until the solution was saturated. The organic layer was removed; the aqueous layer was extracted with ether; the combined organic layers were dried and filtered; and the filtrate concentrated to X-8951A -37- 122092/2 give an oil. The crude oil was purified by flash chromatography on silica gel eluting with 10% EtOAc/hexanes and increasing the polarity to 20¾ EtOAc/hexanes after about 1.5 liters of solvent to yield 9.53 g of a light yellow oil (65% yield) . MS.

To an anhydrous 0°C ether (150 itiL) solution of the above alcohol was added triethylamine (2.93 g, 28.91 mmols, 1.5 eq.) followed by dropwise addition of mesyl chloride (3.31 g, 28.91 mmols, 1.5 eq.) with vigorous stirring. After 3 hours at 0°C, TLC (10% EtOAc in hexanes) indicated the starting material was consumed. The reaction was diluted with ether, washed with water, brine, dried over MgS04, and the solvent removed. The resulting oil was passed through a pad of silica eluting with 25% EtOAc/hexanes, and the eluant was concentrated. To an acetone (200 inL) solution of the resulting oil was added NaHC03 (0.17 g, 1.93 mmols, 0.1 eq.), and Nal (28.88 g, 192.7 mmols, 10 eq.) . After stirring 30 minutes at room temperature under a nitrogen atmosphere, the reaction was heated to 50 °C with a water bath. After 2.5 hours, TLC (10% EtOAc in hexanes) indicated that the mesylate was consumed. The reaction mixture was diluted with ether (500 mL) , washed with cold saturated aqueous Na2S03, water, brine, dried (MgS04), and the solvent removed. The resulting oil was passed through a pad of silica eluting with 5% EtOAc in hexanes to give the purified title compound 10.3 g as a colorless oil (85% yield) .

Preparation 6 3-bromopropyl acetate 3-bromopropan-l-ol (0.54 mole , 75 g) in CH2CI2 (500 mL) at 0°C under N2 was treated with acetyl chloride (0.5 mole, 40.2 mL) . To this solution was added triethylamine (0.54 mole, 75 mL) in portions (5 mL) slowly by syringe. The reaction mixture was allowed to gradually (12 hours) come to room temperature. The precipitate was filtered off, and the filter was washed with CH2CI2. The filtrate washed with water (2x) , brine (2x) and dried over a2S04, and filtered. The filtrate was concentrated to give the titled acetate 91 g (93% yield) as an oil. MS X-8951A -38- 122092/2 Preparation 7 N- (3-acetoxypropyl) -indole To a stirred 0°C DMF (400 mL) suspension of NaH (60% in mineral oil, 0.705 mole, 28.2 g, 1.5 eq.) in a three-neck flask fitted with a reflux condenser and an addition funnel was added a DMF (150 mL) solution of indole (55 g, 0.47 mole) dropwise. After 30-60 minutes, a DMF (50 mL) solution of the alkyl halide, 3-bromopropyl acetate (170 g, 0.94 mole) was added. The reaction was heated at 50°C for 6 hours and then stirred at room temperature for 5-15 hours.

The solvent was removed in vacuo. The residue was partitioned between CH2CI2 and water. The organic layer was washed with IN HCl (3x), water, brine, dried over a2S0 , and filtered. The filtrate was concentrated to give the titled alkyl indole 102 g as an oil which slowly crystallized. MS Preparation 8 N- ( tert-butoxycarbonyl) -indol-3-yl-acetic acid To a stirred acetone (800 mL) solution of indole-3-acetic acid (26.25 g, 0.15 mole) was added cesium carbonate (48.9 g, 0.15 mole) followed by allyl bromide (15 mL, 0.17 mole, 1.16 eq.). After 12 hours the solvent was removed. The residue was partitioned between water and CHCI3. The organic layer was washed with brine, dried over Na2S0 , and filtered. The filtrate was concentrated to give the allyl ester 27.9 g (74% yield) as an oil.

To an acetonitrile (500 mL) solution of the allyl ester (27.9 g) was added di-tert-butyl dicarbonate (29.1 g, 0.133 mole , 1.2 eq.) and 4-dimethylaminopyridine (1.36 g, 0.011 mole , 0.1 eq.). After 15 minutes, the reaction mixture was diluted with EtOAc (1.2 L) and washed with 0.1 N HCl, water (2x) , and brine (2x) . The organic layer was dried over a2S04, filtered, and concentrated to give the BOC protected ester (32.9 g, 94%) as an oil which slowly crystallized.

To a CH2Ci2/EtOAc'i0:3 (325 mL) solution of the BOC protected ester was added sodium 2-ethylhexanoate (17.3 g, 0.104 X-8951A -39- 122092/2 mole ), triphenylphosphine (4.93 g, 18.8 mmol, 0.18 eq.) and ?d(PFh3)4 (4.56 g, 3.95 mmol, 0.04 eq.). After 1 hour, the solvent was removed. The residue was partitioned between EtOAc and water. The basic aqueous layer was back extracted with EtOAc, then ether, and then carefully acidified with 0.10 N HCl. The acidic aqueous layer was then extracted with EtOAc. The organic layer was washed with water, brine, dried over a2S0 , and filtered. The filtrate was concentrated to give the BOC protected acid (21.8 g, 77% yield) as an oil which slowly crystallized. The yield of the titled compound was 53% over three steps. MS Preparation 9 (±) 3, 4-[l, 1 (3-tert-butyldiphenylsilyloxymethyl) exano] -bis- (3-indolyl) ]-l- methyl -lH-pyrrole-2, 5-dione A DMF (50 mL) solution of 3, 4-bis- (3-indolyl) -1-methyl-lH-pyrrole-2, 5-dione (3.41 g, 10.0 mmol) containing the dibromide 3- tert-butyldiphenylsilyloxynethyl-1, 6-dibromohexane (5.64 g, 11 mmol, prepared in a manner analogous to the benzyl derivative in Preparation 3) was added using a syringe pump over a 15 hour period to a DMF (350 mL) slurry of CS2CO3 (11.2 g, 34.3 mmol) at 60 °C. After 4 hours from completion of the addition, the reaction was cooled to room temperature, poured into water (1.5 L) , and extracted with CH2C12 (3 x 300 mL) . The organic phase was washed with water, dried, filtered and concentrated. The concentrate was purified by flash chromatography eluting with 10% to 25% ethyl acetate/hexane to give the macrocycle 3, 4- [1, 1 ' -[(3-tert-butyldiphenylsilyloxy-methyl) hexano] -bis- (3-indolyl) ] -1- methyl -lH-pyrrole-2, 5-dione 2.95 g (43% yield) as a red oil. MS Preparation 10 (S) -methyl 4-tert-butyldiphenylsilyloxy-3- (allyloxy) butyrate To a cyclohexane (400 mL) solution of (S) -methyl 4-tert-butyldiphenylsilyloxy-3-hydroxybutyrate (20.0 g, 53.7 mmol) was added allyl trichloroacetimidate -(21.74 g, 107.4 mmol), followed by trifluoromethanesulfonic acid (1 mL, 50mL/g alcohol) X-8951A -40- in five portions over 30 minutes, with stirring under a nitrogen atmosphere. After 70 hours, the solids that formed were filtered, and the filter cake was washed with cyclohexane, and the volatiles were removed in vacuo. The resultant oil was placed on a plug of silica and washed with hexane, and product eluted with 10% ethyl acetate/hexane . NMR indicated the presence of residual imidate (ca. 10%) ; however the material was carried on without further purification. The residue yields 24.76 g of material, of which approx. 22.2 g was desired product (100%) . MS.

Preparation 11 (S) -4-tert-butyldiphenylsilyloxy-3- (2-iodoethoxy) -1-iodobutane DIBAL-H (231 mL, 1.0M in toluene, 231 mmol) was added dropwise over 40 minutes to a solution of (S) -methyl 4-tert-butyldiphenylsilyloxy-3- (allyloxy) -buty*rate (23.8g, 57 mmol) dissolved in anhydrous THF (1.0 L) at -75°C under N2. After stirring 1.5 hours, the mixture was allowed to warm to -10°C and quenched with 5% water in methanol and a large amount of Celite. The quenched reactionimixture was filtered through a pad of Celite; the filtrate was concentrated and partitioned between ether and 20% citric acid. The ether layer was dried and concentrated in vacuo. The residual oil was passed through a pad of silica eluting with chloroform to yield 20.6 g (93%) of (S) 4-tert-butyldiphenylsilyloxy-3-allyloxy-butan-l-ol.

To a methanol (500 mL) solution of (S) 4-tert-butyldiphenylsilyloxy-3-allyloxybutan-l-ol (20.6 g, 53.6mmol) was added ozone at -78 °C for approximately 12 minutes. The reaction mixture developed a faint blue color, NaBH4 (12.2 g, 321 mmol, 6 eq.) was added to the reaction vessel. The reaction was allowed to come to room temperature . The volatiles were removed in vacuo. The residue was passed through a plug of silica eluting with ethyl acetate to yield 16. g (79%) of (S) 4-tert-butyldiphenylsilyloxy-3- (2-hydroxy-ethoxy) -butan-l-ol as a colorless oil.

To an ether (G00 mL) solution of (S) 4-tert-butyldiphenylsilyloxy-3- (2-hydroxy-ethoxy) -butan-l-ol (15.7 g, 122092/2 X-8951A -41- 40.4 mrnol) at 0°C under nitrogen was added triethy1amine (16.8 mL, 121 mrnol) followed by mesyl chloride (9.38 mL, 121 mmol) . After 3 hours, the solution was filtered; the filtrate was washed with water (2x) , brine (2x) , dried over Na2S0 and concentrated in vacuo. The residue gave 21.9 g (>99%) of the bismesylate as a yellow oil which was carried on directly. The bismesylate was dissolved in acetone (1.4 1), which had been distilled from potassium carbonate. To this solution was added Nal (90.4 g, 603 mmol) and 0.05 eq. NaHC03 (170 mg, 2mmol) . The reaction mixture was kept at 56°C for 24 hours and filtered; and the filtrate was concentrated in vacuo. The residue was partitioned between ether and 10% a2S03, the ether layer was washed with brine, dried over a2S0 , and concentrated to give 17.9 g (73.2%) of (S) - (4-tert-butyldiphenylsilyloxy) -3- (2-iodoethoxy) -1-iodobutane as a colorless oil. The overall yield was 54%. MS: MW= 608.39; observed: 559 (M- tertbutyl; FD, CHCI3) .

Preparation 12 (S)-3,4-[l,l' - [3-oxa-4- (methylsulfonyloxymethyl) hexano] bis- (3- indolyl) ] -lH-pyrrole-2, 5-dione 3, -bis- (3-indolyl) -lH-pyrrol-2, 5-dione (10.04 g, 29.4 mmol) and (S) -4- (tert-butyldiphenylsilyloxy) -3- (2-iodoethoxy) -1-iodobutane (17.9g, 29.4 mmol) were combined and dissolved in anhydrous DMF (80 mL) . The solution was added via syringe pump addition over 72 hours to a suspension of cesium carbonate (38.3 g, 118 mmol) in anhydrous DMF (1.7 L) at 50°C under 2 · The DMF was removed in vacuo. The residue was partitioned between CHCI3/IN HC1. The acidic layer was back-extracted with 122092/4 -42- chiorofora nd ethyl acetate. The combined organic layers wars was ed with IN KCI fix), water (2x), brine (2x) dried over 2S04, and reduced to give a magenca solid. The crude re ction mix re was used without further purification.

The crude reaccior. mixture was suspended in ethanei { 7CC mL) and treated with 5N KOK (800 mL) . The reaction temperature was raised to.80°C. Af er 72 hours the ethaneI was removed in vacuo; the aqueous suspension was cooled to 03C, and acidified with 5M HCl. The violet precipitate was collected and passed through a silica plug eiuting with ethyl acetate. The eluant was concentrated -to- yield 8.7 g of the partially silyiated 2, 5-furandione as a magenta solid that was carried on to the next reaction, without further purification.

: To a DMF (1 Li solution of the above 2 , 5-f randione {8.7g, lS.7amol} was added 1, 1, 1, 3, 3, 3-hexamethyldisila∑ar.e (41.6 mL, 1S7 mmol) and methanol (4 sL, 93.5 tcaiol) under nitrogen at ambient temperature. After 40 hours/ the reaction was concentrated in vacuo , a 2:1 (v/v) MeCN/l HCl solution {100 mL) was added. The residue was stirred for one hour. The organic solvent was removed; and the aqueous suspension was extracted with ethyl acetate. The solvents were removed to yield 8.9 g of maleimide that was used without further purif ca ion.

To a CE2CI2 (800 mL} suspension of the above maleimide (8.9g, 20 mmol) under nitrogen at ambient temperatures was added pyridine (4.85 ml, 60 mraoi) and a slight excess of ae hanesulfor.ic anhydride (4.21 g, 24 mmol) . After 16 hours the reaction mixture was washed with 0.1N KCI, brine, and the organic layer was concentrated. The residue was passed through a plug of silica eiuting with a slow gradient of 0-10% MeCN in CK2 I2 · The eluant fraction containing ' the desired mesylate was concentrated to yield 2.8 g of the title compound as a magenta solid. Overall yield from the diiodide is 13%. MS: M =520; observed 520 JsD, CHCI3! .

Preparation 13 3- ( tsr -butyldlohenylsilyloxyasthyl) -'--cyclohexer.e X-3351A -43- ϊα a mixture of 3-cyciohaxene-i-aethanoi (Aidrich, ' 13.0 mL, O.Li moi) , ^ ¾"- Preparation 14 . 3- ( tert-butyldiphenylsilyloxyaathyl) -1., 6-hexanediol Ozone was .bubbled throug a well-stirred solution of 3- (tert-butyidiphenyisilyloxyaethyl) -1-cyclohe-sene, (13.0 g, 5l.3.mia.oi) in CE2C12 (550 mL) at -78°C until the blue.color of ; unreacted ozone persisted. Tha reaction mixture was allowed to warn.- .to 25° C. Dry.N2 was bubbled .through the solution for 30 minutes. . Bora-ne-diaethyisuifice complex (10.0 M, 23 mL, 0.23 moi) was added dropwise over 10 minutes. The mixture was '-slowly stirred under N2 at 25" C for 24 hours. 5¾ HCi (15 mL) was added, and the' eaction mixture was stirred for 1 hour. Solid EC03 was added, until tha mixture tested basic to pS paper..

After -filtration, the filtrate was washed with 200 mL portions of 5% Na≤C03 and water and dried over anhydrous MgSGd After ■ evaporation of the solvent under reduced pressure, the crude product was purified by chromatography through a 4" x 4" pad of silica gal eluting with EtQAc.' 3-(tsrt- .butyidiphenyisiiyioxy methyl) -1, β-hexanadioi 17. S g (SQ%) . was obtained as a colorless viscous oil which was homogeneous by TLC (Rf U.5, ether) .

Analytical calculated for C23¾403Si-(Q .2 Ξ20) :" 122092/4 -44- C, 70.90; H , 3.83.

Found: C, 70.72; H, 3. So.

Prepara- ic 15 3- i teri-butyldiohsnylsli yloxyrnethyl) -1, 6-dicrorr,ohs: Preparacion 16 (S) - (-) -3-Cyclohexene-l-methar.ol A solution of LiAlH4 (1.0 in TKF, 75.8 ciL, 75.S .TJT.OI) was added dropwise ever 15 .Tiinutes co a cooled soiu-rion cf che known compound (Ireland et ai J. Org. Chem. 1992, 57(19), 3071-5073 and references therein), 3-( 3—Cyclohexene-i-carbonyloxy) -l-methylpyrrole-2, 5-dione, (8.23 g, 34.5· X-8S31A -45- rnci) iii THF ISO mL) . The reaction mix u e ' as allowed to wars to roos. temperature and stirred at 25 C for 2 hours, cooled quenched wi h water ' and I aQE. The. mixture was filtered throug Celite. The solids were washed with ΤΕΞ" (100 mL} . ,5. A ter evaporation- of the filtrate, u der reduced pressure, the residue was dissolved into 150 mL qf ether and washed, with water . (2 x 50 mL) and brine (30 'mL) and. dried over anhydrous MgS04.

E apo ation of the solvent gave . (S) ■- (-) -3—Cyciohexene-I-mathanol ■ 3.24 g -(S3%) as a clear oil (a]D .= -90:3 (C = I, CE3CE) . 3oth 0 the TIC properties' and lE . SR spectrum of this material were identical in all respects with .that of the racemic' material • (Aldrich) .

XS OMR .(300 MHZ, CDCl3),5l.2I - 1.42 ( , 73.) , 1.63. 1.83 (m, 3S), 2.04 - 2.21 (a, 3Ξ) , 3^5 (brs, 2E) , 5.69- (s, 2S) , 5 ·' ■ · ■ ■.

:- Preparation .17.

(S) - (-) - 1- (tertbutyldiphenylsllyloxymethyl) -1-cyclohexene ; (5).- (-) -3-Cyclohexene-l-methanol 3.17 g, 28.3 mmol) . was treated with tart-bu yidu^phenylcniorosilane (8.15 mL,- 31.1. 0 mmol], N,N- ciisopropyletnylamine . (10.9 mL, .62.3 mmol} and - dimethyiaininopyridine. (1.Q3 g, 8.5. mmol) in.CE2Cl2 (1Q0 xaL) to ■ afford, afte workup and chromatography, silyl ether (Sj-(-)- ' 3- (tert±)utyldiphenyisiIyioxymethyl}--l-cyclohexen'e 8.73 g (.83%) . as a clear oil: . Both the TLC properties and -E fcS. "spectra of ■ 5 . this material were identical in all respects with racemic silyl · ether 3- (tertbutyldiphenylsil loxymethyl) -I-cyclohexene .

-S MMR (300 iH , C C13151.05. (s,9E) , 1.29. (m, IE) , · 1.71-2.13 ( , 4H) 3.54 (d, 2E, J = βΕζ) , 5.66 (hr s, 2E) , 7.33 and 7. So (m, 0 10 S) .

Pre aration · 18 (5) - (-) - 3- (tart-butyldiphenylsilyloxymathyl) -I, 6-hexanediol F llowing .the same, procedure described for the 5 preparation of the racemic dial 3- { tert-butyidiphenylsiiyloxy- methyli -L, S-aexanedioi , silyl ether (3) - (-) - 3-tert-butyldi- X-SS51A - 6- phenyisilyioxyaathyi} -1-cyciohe ana (3.35 g, 23. S aaci) was ozonized, then rs uctivaly war-ad-up. (3≤3-½e2S) cc afford' (5)- (--) -3-{tert-butyIdipher,yLsilyIaxyaethyi} -1, S-hexanediq1 - 5.01 g (53%) as'a colorless viscous ail, which was homogeneous by TIC ( f = 0.4 E CAc) .. ■ Τ-Ξ NH (300 .Hz, C2CI.3 J 51.05 _ (s 5Ξ) , 1.21-1.81 (a, 7H) , 2.32 (br s, 2H) , 3.3C-3.73 (a, 6Ξ) , 7.3.2 and 7,7Q (a, I0H) . ' Preparation 13 (5} -3- (tgrt-butyldlphsnylslly oxyaethyl) -1, 6-dlbreachexana Following the saae .procedure describe, far tha prep-' araticn- of raceaic . d? bromide, 3- (tart-but idiphan i ilyicxy-aethyl) -1, 6-dibroaohexana, (S) - {-) -3- (tertbutyldipheaylsilyioxy-.methyl) - S-haxanedial {4.85 g, .12.53 macl) was reacted with fr-brcaosucciniaida (5.33 g, 30.1 aaol) and tripheryiphosphina (7.37g,3Q.i aaol) in C¾ i2 ( SO aiL) at 0°C to afford compound (S) - (-) -3-{tert-butyldiphaj3.ylsiiyicxyaethyl) -1, S-dibroaahexana 4.81 (75%} as a clear, colorless oil which was hoaogeneous by TLC (R-f = 0.8, 10% EtOAc in hexanas . 3ath the TLC properties and -E spectra of this compound ware identical in ail respects with raceaic isomer.. HS . s NMB. (300. MHz, CDCl3)5l .0S (s, 9Ξ) , ' 1.35 - 2.10 (a, 7H) , 3.55 (a, 4Ξ) , 3.56 (app d, 2E, J = 4Ez) , 7.40 and'7.64 (a, 10E) .

Preparation -'.20 (R) - 3- ( tert-butyldiphsnylsilyloxyaethyl) -1 , S-dibroaohexane Following tha same procedure described for tha . preparation of (S) -(-) -3- (tert-b tyi-a.phenyisiIyioxyaethyi) -1, S-dibromahaxane, (5) - (-)- 3- ( tert-butyidiphanyisiiyloxy- ' aathyi) -1, S^haxanecioi. (5.05 g, 13.04 aaol) was raacta with N-broaosucciniaide (5.57g, 3 .32' aaol) and triphenylphcsphina :. (3.21 g, 31.32 aaol! i CH2C12 (150 aL) at OcC to afford chirai dibroaide (?,)- 3-.( tert-bu yidi?hsnyisil io aa hylΐ -1,6-dibrcmohaxana, 5.33c, (87%) as a clear, colorless :oii which was homogeneous by TLC (Rf =-Q.S 10% E OAc in hexanes. MS .

X-S951A -47- IE- NHR' (300 ΜΞζ, CDCl3)51.q6 (s, SE) , 1.35 - 2.10 (m, 7Η) , 3.55 (Π./4Η)/ 3.56 (app d< 2Ξ, J = 4 Ε2) ,' 7.40 and 7.64 (a, 10Ξ) ..

Preparation. 21 2-allyl-4-pei-.ters.oic acid To a stirred suspensio of sodium methcxide (59.4 g, 1.1 moi). in dry methanol (1 L) at G°C- was added dimethyImaionate (.57 L,-0.-5 mol) dropwise . under N2- After 30 minutes, aliyl- bromide (S5.mL, 1.1 moi) was added in one portion. Afte 14 hours, at ambient, temperature the reaction was concentrated in . vacuo. . The residue dissolved in methanol (0.5 L) and treated with 5N NaOE (500 mL).. After stirring for 24 hours, the methanol was removed in vacuo, and the basic aqueous layer washed. with ethyl acetate (2X) - The aqueous layer was acidified ; with'SN HCi (0.5 L) and extracted with ethyl" acetate.'.. The organic extract was washed with water C2x) , brine, dried over N 2S04, and concentrated 'in . vacua to. a .white solid. Trituration o . the resulting solid with pantane and atmospheric drying' gave 51.4 g (57% yield) of the diacid._ The diacid (50g, 27.4 mmol> was heated ■ (,i50'°C) until -COi .evolution .ceased (about 2 hours') . : " The residual brown oil was eluted with ethyl acetate through a ;, small silica plug to yield the title compound 32.8g (85%) as a golden oil.- The overall yieid'for the. three steps is 48%. -H NMR: (CD3C ) δ '2.4.-· (m, .4E) ; 2.5 (m, - 1H) ; 5.05 (dd, 2Η)·; 5.15 . (dd, 2Ξ) ; .5.9 ( , 2Ξ) ; 12.8 (br, 1Ξ) . MS.

Preparation .22 3- (tertbutyldiphenylsilyloxymethyl) -pentane-1, 5-diol To a 0°C stirred suspension .o-f LAS (4.33g, 1.14 mmci) in anhydrous ether (125 mL) was added 2-aiiyl-4-pentenoic acid .■(16.0 g, 114 mmoi) dropwise under N2 - The reaction . mixture was allowed to come to room 'temperature . After 16 hours, the reaction was quenched with ethanol (25 mL) followed by 4N ECi (.40 mL) , extracted, extracted with ether (2x) , dried', and concentrated in vacuo to give the alcohol,, 2-aiiyi-4-penten-l- ■ol, as a colorless oil 11. g (82%) that was used without further purification. ' X-8951A -48- 122092/2 To a dry CH2CI2 (0.5 rtiL) solution of 2-allyl-4-penten- l-ol (11.7 g, 93 mmol), was added imidazole (12.6 g, 185 mmol) followed by chloro tertbutyldiphenylsilane (25.48 g, 93 mmol), and stirred for 16 hours. The reaction was filtered, the filtrate was washed with water, brine, dried and concentrated in vacuo to give the silylether, 3-(tert- butyldiphenylsilyloxymethyl) -1, 6-heptadiene 32.5g(96%) as an oil that was used without further purification.

Ozone was bubbled through a -78 °C dry methanol (500 mL) solution of 3- (tert-butyldiphenylsilyloxymethyl) -1, 6-heptadiene (17 g, 47 mmol) until a blue tint persisted (30 minutes). The reaction was purged with nitrogen (20 min.) and NaBH4 (17.6g, 47 mmol) was added. The cold bath was removed and the reaction brought to room temperature. The reaction was concentrated in vacuo and the residue partitioned between ether and brine. The ether layer was concentrated and the residue eluted over a silica plug with 0-50% ethyl acetate/hexanes. The minor component was pooled and concentrated to yield the diol , 3- ( tertbutyldiphenylsilyloxymethyl) -pentane-1, 5-diol, 3.8g (22%) of the desired diol as a colorless oil. Overall the yield for the three steps is 17%. MS.

NMR: 6 1.17 (s, 9H) ; 1.6 (dt, 4H) 1.83 (m, 1H) ; 2.14 (s, 2H); 3.6 (m, 6H) ; 7.41 (t, 4H) ; 7.45 (t, 2H) ; 7.66 (d, 4H) .

Preparation 23 1, 5-diiodo-3- (tert-butyldiphenylsilyloxymethyl) -pentane To a 0°C ether (300 mL) solution of 3- ( tert-butyldi-phenylsilyloxymethyl) -pentane-1, 5-diol (6.9g, 19 mmol) was added methanesulfonyl chloride (4.3 mL, 56 mmol) followed by Et3N (7.7 mL, 56 mmol) . After 3-16 hours, gradually warming to ambient temperature, the reaction was washed with water, brine, dried over MgS04, and concentrated to give the 1,5-bis (methanesulfonyloxy) -3- ( tert-butyldiphenylsilyloxymethyl) -pentane 8.5g (90%) as a colorless oil that was used without further purification.

X-8951A -49- o a freshly distilled acetone (500 mL) solution of the bis-mesylate, 1, 5-bis (methanesulfonyloxy) -3- ( tert-butyldi- phenylsilyloxymethyl) -pentane, (8.5g, 16 mmol) was added excess Nal (36. lg, 241 mmol) and NaHC03 (67 mg, 0.8 mmol). The reaction was refluxed (57 °C) for 72 hours, cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was diluted' with ether, washed with 10% Na2S03, dried, and concentrated to give the title compound 7.4 g (78% yield) as a colorless oil. The overall yield for two steps is 70%. MS.

!H NMR: (DMSO-d6) δ 1.06 (s, 9H) ; 1.78 (m, 1H) ; 1.8-2.06 (m, 4H); 3.13 (m, 4H) ; 3.57 (d, 2H) ; 7.38-7.46 (m, 3H) ; 7.64 (d, 2H) .

Preparation 24 2- (2 ' -Bromoethoxy) -benzylbromide Ozone was bubbled through a -78°C dry methanol solution of 2- (allyloxy) enzyl alcohol (LaChapelle et al Tetrahedron, 44(16), 5033-5044 (1988)) (7.0 g, 43 mmol) for 13 minutes, checking the teaction TLC profile every 2 minutes for complete disappearance of the starting olefin (Rf=0.8, 75%EtOAc/hexane) . The reaction mixture was purged with nitrogen, NaBH4 (9.7 g, 0.25 mol) was added and the reaction temperature brought- to 0° C. After 30 minutes, the reaction was warmed to room temperature, concentrated, diluted with ether, washed with water, brine, dried and concentrated to a residue. The residue was eluted through a pad of silica with EtOAc/hexanes (gradient elution 25% - 75% EtOAC) . Evaporation of the eluting solvent gave the diol, 2- (2 ' -hydroxyethoxy) - benzyl alcohol, (4.8 g, 67%) as an oil. .MS: MW = 168; observed •168, FD, CHC13) · T.o a 0°C dry CH2C12 (250 mL) solution of the diol, 2- (2 ' -hydroxyethoxy) -benzyl alcohol, (4.38 g, 26 mmol) was added triphenylphosphine (15.8 g, 60 mmol) and N-bromosuccinamide. (1.0.7 g, 60 mmol). After 2 hours at 0° C, the reaction was complete by TLC (20% EtOAc/CH2Cl2) analysis, and the reaction was concentrated in vacuo. The concentrate was eluted (hexane - 15% X-8951A -50- 122092/2 EtCAc/hexane gradient) through a pad of silica gel . Concentration of eluting fractions gave the dibromide, 2- (2 ' -Bromoeth- oxy) -benzylbromide, ' (6.91 g, 90% yield) as a colorless solid. MS. 13C-NMR (CHC13/ 75.4 MHz) δ 28.7, 29.1, 68.2,, 112.3, 121.6, 126.8, 130.2, 131.1, 156.0. 1H-NM. (CHCI3, 200 MHz) δ 3.72 (2H, t, J=5 Hz), 4.34 (2H, t, *J=5 Hz), 4.59 (2H, s), 6.84 (H, d, J=7 Hz), 6.95 (H, t, J=7 Hz), .25 - 7.38 (2H) .

Preparation 25 1- (tert-butyldimethylsilyloxy) -3- (2-iodoethoxy) -4- (tert- butyldiphenyl) -butane The allyl ether, 1- (tert-butyldimethylsilyloxy) -3- (allyloxy) -4- (tert-butyldiphenyl) -butane, (21.6 g, 43.4 mmol) was dissolved in methanol (500 mL) and cooled to -78 °C under nitrogen. Ozone was bubbled into the reaction and after 11 minutes it was judged complete by TLC (9 hexane/1 ethyl acetate) . Sodium borohydride (9.9 g, 6 eq) was added and after 5 minutes the reaction was allowed to warm to room temperature. The methanol was removed in vacuo. The residue was suspended in ether (800 mL) . The ether was washed with water, and the aqueous phase backwashed with ether. The combined organics were washed with brine, dried (Na2S04), filtered and concentrated in vacuo to give an oil. The material was passed through a silica pad with 5% ethyl acetate/hexane followed by elution of the product with 25% ethyl acetate/hexane to provide 11.0 g (50% yield) of the alcohol, 1- (tert-butyldimethylsilyloxy) -3- (2-hydroxyethoxy) -4- (tert-butyldiphenyl) -butane as a light yellow oil. MS. NMR.

To an anhydrous ether (200 mL) solution of the alcohol, 1- (tert-butyldimethylsilyloxy) -3- (2-hydroxyethoxy) -4- (tert-butyldiphenyl) -butane, (11.0 g, 21.9 mmol) under nitrogen at 5 °C was added triethylamine (4.6 mL, 1.5 eq) and methanesulfonyl chloride (2.5 mL, 1.5 eq) . After 1.5 hours the reaction was complete by TLC (5% ethyl acetate/dichloromethane) . The reaction was diluted with ether (250 mL) , washed with water X-SS3LA -51- (2X) , brina , dried (N ^SGd) , filtered and concentrated is acrc to gi~e an cil. ~ a naterial was passed though a silica pad aiutirc with 5%. ethyl acatata/haxana followed by 22% ethyi acata e/ha-xare to prcvida II. S g (91% yield) of tha nesylata,. 1- (tarn-b cyldinethyisiiyic y) -4 - ( cart-bucyldlph.ar.yl) -butane as an ail. as. bOR.

To an acetone (3GG stL) solution of tha nesylata, 1- { ert-butyldinathylsiiyioxy} -3- (2- (na ihanasulfcriylcxy} ethcxy) -4- " (tert-b tyldiphenyl) -butane, (II.6 g, 20 SEC I) under nitrogen - was added sodi m iccica (44 q, IS ac) arc secies bicarbonate · . (I7G sg, G.i ac]- . Tha ni tura was rsfiuxad for 13- hours · followed by rasovai af tha acetone In vacua. The resulting residua was suspended in ether, washed with, water (2 ) , and tha aqueous phase bacrwashad with .atha . Tha co-abided atrar portions - vers washed with 1Q'% sodiu-s. sulfite solution., brina {IX) , dried -( qSQd) , fii ared and concentrated s V2 sa to provide IG.7 g (S-7¾ yialc) of the title iodica as an cil which, was used without f'irther purification. 02. 1£E..

Preparation 25 . 1- (2- (nathylsulfogyloxy) -athoxy) -2- ( (riat ylsul onylcxy) ethyl! -3- . . ( tart-butyldlphanylsilyloxy) -propana To a stirrad solution of dirieth l ailyl Etaianata (.3 q,. 0.2 no!) in t-butyi alcohol (0.5 L)' was solid". so di us borchydrlda (13 q, 0.3 JEOI.) . The reaction was heatad (70. "C) and satharcl (152 aiL! was added dropwisa over a period of I hour.. Tha nixtura ■ was stirrad overnight at rooa. ter-peratura .

?Tata . (20 nL) was added to. da stray the excess barohydride. Tha "resulting mixture was' filtered through Caiite. Tha filtrate 'was concentrated (IQG al) , and extracted with ethyl acatate (20 SLL X 4) . Tha combined attracts were dried eve ifcSCd and concentrated under reduced pressure to afford relatively clean dicL, 2- aiiyLpropane-l, 3-dicI, (IS g, 33%, yialc) that was carried ever co the nejct reaction without any f rither purification.

To a stirred solution of dial, 2- (2-prcper-i- yi) propane- I, 3-dioi, (23.2 q, 0.1S ECU in taiuana (I L) was. added anis aldehyde (27.3 , Q.20 no I) and pyridinium p-toluene Sulfonate (4 g, IQ .

X-3551A -52- ' moi¾) . The flask was equipped with, a Dean starl trap, a_ud the reactio . mixture was refiuxed. After 5 hours, the reaction mixture was cooled to room temperature, diluted with ether (1 L) , washed with sat. ffs≤C03 (20 mL x 3), -water ''{5G mL x 3), and brine (30 mL) . The organic layer was dried ever MgSGa, and ' conce traced under -reduced pressure to give a residue. The residue was eiuted through a short silica gel column with 10% ethyl acetate in hexane and evaporation cf · the eiuting solvent . gave the. anlsyiidene, 1, 3-0-anisyiicane-2- (2-propen-i-yi) propane . (40 g, 89%). ■ (Rf = 0.62 (25% ethyl acetate in hexane) ) To a stirred mixture of the anisylidene 1,3-G- anisyiidene-2- (2-propen-l-yi) propane (20. G g, 83.3 mci) in CH2C12 (500 mL) and. ρΞ 7.0, buffer (25 mL) 0' C was added BJJQ (33.7 g, I7Q.7 mmoi) - The- reaction mixture s stirred. vigorously and allowed to warm up to room temperature. Afte 12''. ■ hours, .the reaction was diluted with ether (1L), washed'.with ' . sat. aq. NaHCG3 (200 mL X 2), and 1Q¾ aq. Na2SO-3 (200 mL X 3) , . dried, and concentrated under reduced pressure to a residue.

The residue was eiuted through a silica gel- column with ethyl cetate/hexane (10% - 25%. ethyl acetate gradient) and evaporatio of the. eiuting. solvent gave the anisoyloxy . . alcohol, 3-<4-methoxybenzpyloxy ) -2_- (2-propen-l-yi) -propah-l-oi (12.7 g, 61%) . (Rf .= 0.14 (25% ethyl acetate in hexane) . *E£ . .

To a stirred solutio of alcohol, 3-(4-- methoxybenzoyloxy) -2- (2-propen-l-yi) -propan-i-ol, (16.53 g, 65.32 ■ . moi} in CH2 12 (250 mL} was Γadded .0-allyl trichloroacati idate (24.30 g, 132.64 mmol) in cyclohexane (500 mL) .; To this mixture was added trifluoroacetic acid ..(1 mL) under a N2 atmosphere. After .12 hours a white precipitate had formed. The reaction was filtered. The filtrate was diluted with ether (500 mL) , washed with water (100 L 3), and brine (100 mL) , dried, and concentrated under reduced pressure to a residue. The residue was eiuted through a silica gel column with ethyl acetate/hexane (0% - 25% echyi acetate gradient). The diene, 1- (2-prapen-l-yi- cxy) -2- (2-propen-i-yi) -3-( -msthcxybenzayloxy). -propane. (24 g). containing some acatamide was taken to next step without any further purification. ( f = 0.33 (25% ethyi acetate in hexane) X-8951A -53- 122092/2 The ester, 1- (2-propen-l-yl-oxy) -2- (2-propen-l-yl) -3- (4-methoxybenzoyloxy ) -propane, (24 g)was dissolved in THF (60 mL) and methanol (100 mL) and IN aqueous NaOH (40 mL) was added.

The resulting mixture was stirred overnight followed by removal of methanol and THF under reduced pressure. The concentrated reaction mixture was diluted with ether (250 mL) , extracted with ether (100 mL x 3), dried, and concentrated under reduced pressure to give a residue. The residue was eluted through a silica gel column with 10% ethyl acetate/hexane and evaporation of the eluting solvent gave the alcohol, 1- (2-propen-l-yl-oxy) -2- (2-propen-l-yl) -propan-3-ol (4.10 g, 30% for 2 steps). NMR.

Rf = 0.23 (25% ethyl acetate in hexane) .

To a stirred CH2CI2 (150 mL) solution of the alcohol, 1- (2-propen-l-yl-oxy) -2-(2-propen-l-yl)-propan-3-ol (4.10 g, 26.2 mmol) was added imidazole (2.70 g, 39.7 mmol) under a 2 atmosphere. After the imidazole had dissolved tert-butylchlorodiphenylsilane (8.24 g, 29.97 mmol) in CH2CI2 (50 mL) was added over 10 minutes. After stirring 12 hours, the reaction was diluted with ether (100 mL) quenched with water (100 mL) , and extracted with ether (100 mL x 3) . The combined organic phase was washed with brine (100 mL) , dried, and concentrated under reduced pressure to give a residue. The residue was eluted through a short silica gel column with ethyl acetate/hexane (0% to 25% ethyl acetate gradient) and evapo-ration of the eluting solvent gave the silyl ether, l-(2-propen- 1- yl -oxy) -2- (2-propen-l-yl) -3- (tert-butyldiphenylsilyloxy) -propane (7.41 g, 72% yield). Rf = 0.76 (25% ethyl acetate in hexane).

Ozone was bubbled through a -78 °C methanol (500 mL) solution of diene, 1- (2-propen-l- yl-oxy) -2- (2-propen-l-yl) -3- (tert-butyldiphenylsilyloxy) -propane, (7.41 g, 18.80 mmol). After the disappearance of the starting material (TLC, 25% ethyl acetate/hexane), the reaction mixture was purged with N2 and sodium borohydride (2.13 g, 56.30 mmol) was added. The reaction was warmed to room temperature. After 12 hours, the reaction was concentrated. The white residue was quenched with water, and extracted with ethyl acetate (100 mL x 3) . The combined organic phase was washed with brine, dried, and concentrated X-8951A -54- under reduced pressure to give a residue. The residue was eluted through a short silica gel column with ethyl acetate/hexane (10% to 50% ethyl acetate gradient) and evaporation of the eluting solvent gave the 1,7-diol, 1- (2-hydroxyethoxy) -2- (2-hydroxyethyl) -3- (tert-butyldiphenylsilyloxy) -propane (5.48 g, 72% yield).

Rf = 0.21 (50% ethyl acetate in hexane) . NMR.

To a stirred CH2CI2 ( 400 mL) solution of diol, l-(2-hydroxyethoxy) -2- (2-hydroxyethyl) -3- (tert-butyldiphenylsilyloxy) -propane (5.48 g, 13.6 mmol) under N2 atmosphere was added TEA (11.2 mL, 78 mmol), followed by dropwise addition of methane sulfonyl chloride (3 mL, 39.00 mmol) in CH2CI2 (100 mL) over a period of 30 minutes. After 12 hours, the reaction was diluted with ether (500 mL) , washed with water (100 mL x 3), brine (100 mL) , dried, and concentrated under reduced pressure to a residue, residue was eluted through a short silica gel column with ethyl acetate/hexane (10% to 50% ethyl acetate gradient) and evaporation of the eluting solvent gave the bismesylate, 1- (2^ (methylsulfonyloxy) -ethoxy) -2- ( (methylsulfonyloxy) ethyl) -3- (tert-butyldiphenylsilyloxy) -propane (7.40 g, 97%). Rf = 0.55 (50% ethyl acetate in hexane).

NMR.

X-8951A -55- 122092/2 Example 1 3 , 4- [1, 1 ' - (3-oxapentano) -bis- ( 3-indolyl) ] -lH-pyrrole-2 , 5- dione .

Sodium hydride (60 percent dispersion in mineral oil, 113 mg, 2.82 mmol) was added in portions over 15 minutes to a solution of 3, -bis (3 ' -indolyl) furan-2, 5-dione (337 mg, 1.02 mmol) in 5 mL of dry DMF under Ν2· The mixture was stirred 1.5 hours and then diluted with 5 mL of DMF. Bis (2- bromo-ethyl) ether (0.14 mL, 1.13 mmol) was added dropwise to the green solution. The reaction mixture was stirred for 30 minutes at 25"C and then heated at 50°C overnight. The cooled mixture was poured into dilute aqueous citric acid (75 mL) and extracted with EtOAc (2 x 40 mL) The combined organic extracts were washed with water (3 x 20 mL) and brine (20 mL) , and dried over anhydrous MgS04. The solvent was removed under reduced pressure. The residue was passed through a short column of silica gel (50 percent EtOAc/hexanes) , and then subjected to radial preparative-layer chromatography (Chromatotron) eluting with 50 percent EtOAc-hexanes to afford 82 mg (20 percent) of 3,4-[l,l'-(3-oxapentano) -bis- (3-indolyl) ] -furan-2, 5-dione as a burgundy solid, M. Pt. > 320°C.

A solution of 3, 4- [1, 1 ' - (3-oxapentano) -bis-(3-indolyl)] -furan-2, 5-dione (58 mg, 0.15 mmol) in DMF (1.5 mL) under N2 was treated with a mixture of 1,1,1,3,3,3,-hexamethyldisilazane (0.33 mL, 1.45 mmol) and CH3OH (23 mg, 0.73 mmol) (premixed 10 minutes) . After stirring for 16 hours at room temperature, the mixture was poured into water {20 mL) and extracted with EtOAc (3 5 mL) . The combined organic extracts were washed several times with water, dried (MgS04) and concentrated. The residue was purified by radial chromatography X-8951A -56- 122092/2 eluting with 3 percent CH3OH in CHCI3 to afford 3, 4- [1, 1 · - (3- oxapentano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (41.5 rag, 72 percent) as a violet solid, M. Pt. > 320°C. MS Calculated for C24H19N3O3: 397.1426.

Found: 397.1438. 3, 4- [1, 1' - [3- (hydroxymethyl) -4-oxaheptano] -bis- (3-indolyl) ] - lH-pyrrole-2, 5-dione To a stirred DMF (125 mL) solution of 3, 4-bis- (3- indolyl) ] -l-methyl-lH-pyrrole-2, 5-dione (4.35 g, 12.8 mmol) containing cesium carbonate (8.31g, 25.5 mmol) was added dropwise over 15 minutes a DMF (20 mL) solution of l-(tert-butyldimethylsilyloxy) -3- (3-iodopropyloxy) -4- ( tert-butyldiphenylsilyloxy) -butane (4.0 g, 6.4 mmol) under 2- After 3 hours, TLC (1:1 ethyl acetate/hexane) indicated consumption of the starting iodide. The reaction was diluted with ethyl acetate (200 mL) and washed with water. The aqueous layer was extracted with ethyl acetate (200 mL) ; and the combined organic layers were dried and concentrated. The concentrate was purified by flash chromatography eluting with 10% to 25% ethyl acetate/hexane to give the desired monoalkylated product 3-[l-[3- [1- (tertbutyldiphenylsilyloxymethyl)-3 (tertbutyldimethyl-silyloxy) propyloxy] propyl] -3-indolyl] -4- (3-indolyl) -1-methyl-lH-pyrrole-2, 5-dione 3.94 g (69% yield) as a red oil. MS To a methanol (100 mL) solution of the above alkylation product (3.14 gr., 3.74 mmol) was added toluenesulfonic acid (60 mg, 2%) . After 2 hours, TLC (50% ethyl acetate/hexane) indicated consumption of the starting material. The reaction was concentrated to half the volume, diluted with ethyl acetate (300 mL) , washed with IN NaOH, brine, dried, and concentrated. The concentrate was purified by eluting through a pad of silica with 50% ethyl acetate/hexane to give the desired 122092/2 X-8951A -57- alcohol 3- [1- [3- [3-hydroxy-l- (tertbutyldiphenylsilyloxy) propyloxy] ropyl] -3-indolyl] -4- (3-indolyl) -1-methyl-lH- pyrrole-2, 5-dione 1.76 g (65% yield) as a red foam. MS To a 0°C ether solution (200mL) of the above 3-[l-[3- [3-hydroxy-l- (tertbutyldiphenylsilyloxy) propyloxy] propyl] -3- indolyl] -4- (3-indolyl) -l-methyl-lH-pyrrole-2, 5-dione (1.76 g, 2.4mmol) was added triethylamine (0.5 mL, 1.5 eq) , followed by mesyl chloride (0.28 mL, 1.5 eq) . The reaction was brought to room temperature and was complete after 1 hour. The reaction was diluted with ether (200 mL) , washed with water, brine, dried, and concentrated. The concentrate was passed through a pad of silica eluting with 50% ethyl acetate/hexane, to give the mesylate product which was used immediately.

To an acetone (250 mL) solution of the above mesylate was added sodium iodide (3.6 g, 10 eq) and NaHC03 (20 mg) .

After stirring 4 hours, starting material still existed (TLC, 50% ethyl acetate/hexane) and additional amount of sodium iodide (10 eq) was added, and the reaction was heated at 60 °C. After 4 hours, the starting material was consumed (TLC, 50% ethyl acetate/hexane) . The reaction was concentrated, diluted with ethyl acetate (250 mL) , washed with water, 10% sodium sulfite, dried, and concentrated. The concentrate was purified by passage through a pad of silica gel eluting with 50% ethyl acetate/hexane to give the desired iodide as an oil 3-[l-[3-[1- (tertbutyldiphenylsilyloxy) -3-iodopropyloxy] propyl] -3-indolyl] -4- (3-indolyl) -l-methyl-lH-pyrrole-2, 5-dione 1.71 g (85% yield). MS A DMF (lO L) solution of the above iodide 3-[l-[3-[l- (tertbutyldiphenylsilyloxy) -3-iodopropyloxy] propyl] -3-indolyl] -4- (3-indolyl) -l-methyl-lH-pyrrole-2, 5-dione (2.0 g, 2.4 mmol) was added slowly by syringe pump over 80 hours to a DMF (400 mL) slurry of cesium carbonate (3.12 g, 9.6 mmol). After 3 hours from completion of the addition, TLC (50% ethyl acetate/hexane) indicated consumption of the starting material. The reaction was diluted with ethyl acetate (1 L) washed with water and brine. The aqueous portion was extracted with ethyl acetate (500 mL) . The combined organic layers were concentrated and the concentrate was purified by passage through a silica pad eluting with (50% ethyl acetate/hexane) . Concentration of the eluant gave the desired macrocycle 3, - [1, 1 ■ -[4-oxa-3- (hydroxymethyl) 122092/4 X-3951A -58- heptano] -bis- (3-indolyl) ] -1-methyl-la- pyrrcis-2 , 5-dicne 1.65 g (97% yield) . MS To an ethanol (100 ml) solution of the above N-met vi aaleimide, 3,4- [1,1' [3- (hydroxvTEethvl -oxaheptario J -bis- (3- indolyl) ] -1-nteth.yi-lH- pyrrole-2, 5-dione (1.7 g,2.4jimoi) was added 5N Ofi (50 mL) . After 12 hours, the reaction was heated at 50' C for 2 hours. The reaction was cooled to room temperature, concentrated, diluted with ethyl acetate, and washed with water. The organic phase was dried and concentrated to give the desired, anhydride 3, -[L, 1' -[4-oxa-3- (hydroxymethyl) heptano 1 -bis- (3- indolyl) ] -furan-2 , 5-dione 1.37 g (83% yield) as a red solid. MS To a DMT (100 aL) solution of the above anhydride 3, - I, 1'- [4-OX3-3- (hydroxymethyl) heptano] -bis- (3- indolyl) ] - furan-2 , 5 -dione 1.37 g, 3nmoi) was added 1, 1,1, 3, 3, 3-hexamethyidisiiazane (12.5 ml, 20 ec) and methanol (1.21 mL, 10 eq) . After 24 hours, the starting material had been completely consumed (TLC, 50% ethyl acetate/hexane) . The reaction was diluted with ethyl acetate, washed with IN HC1, water, dried, and concentrated. The concentrate was stirred in IN ECl or with cesiua fluoride to remove residual TMS group.

The reaction was diluted with ethyl acetate, washed with water, dried, and concentrated to give the desired maleimide, 3,4-[1,1' [3- (hydroxymethyl) -4-oxaheptano] -bis- (3-indoiyl) ]-lH-pyrrole-2, 5-dione 1.02 g (75% yield) as a red solid. MS 1H-NMR: (300 MHz in dS-DMSO) : 52.1 (m, 4E) , 2.4 (m, 2K) , 3.2Θ (br, m), 3.4 (a, 1H) , 4.25 (m, 4H) , 4.5 (t, J=6 Hz, IK), 7.0-7.9 (m, 10H), 11.0 (s, IE) 13C-N R: (75 MHz in d6-DM≤0) : 520.9, 28.9, 30.3, 30.9, 34.3, 40.2, 41.6, 42.4, 62.4, 55.9, 78.1, 104.0, 104.1, 110.0, 110.1, 119.6, 11-9.7, 121.4, 121.8, 24.3, 126.5, 126.6, 127.9, 131.5, 131.6, 131.7. 135. a. 135.9. 139.1, 151.4, 172.2 122092/2 X-8951A -59- Example 3 3, - [1, V - [4-oxa-3- (hydroxymethyl) hexano] -bis- (3- indolyl) ] -1H- pyrroIe-2, 5-dione To a dimethylformamide (250 mL) solution of 3,4-bis- (3-indolyl) -l-methyl-lH-pyrrole-2, 5-dione (17.9 g,52.5 mmol, 3 eq) under nitrogen was added cesium carbonate (68.4 g, 4 eq) . To the resulting suspension was added the iodide, l-(tert-butyldimethylsilyloxy) -3- (2-iodoethoxy) -4- (tert-butyldiphenylsilyloxy) -butane, (10.7 g, 17.5 mmol) . The reaction was stirred for 18 hours at room temperature. TLC (5% ethyl acetate/hexane) showed disappearance of the iodide. The reaction was "poured into ethyl acetate (1200 mL) and washed with IN HC1 (400 mL) followed by backwash with ethyl acetate (2X) . The combined ethyl acetate portions were washed with saturated sodium bicarbonate solution, brine (2X), dried (MgS04), filtered and concentrated down in vacuo , bimethylformamide was removed by azeotroping with xylene. The resulting red gum was slurried in dichloromethane and acetonitrile to give a solid suspension. It was concentrated down, more dichloromethane added, cooled and filtered to give a red solid. Some of the desired product was extracted from this solid by another trituration in dichloromethane and then in ethyl acetate. The filtrates were concentrated in vacuo and the resulting residue absorbed on silica and applied to a large flash column. Dialkylated byproduct was removed by elution with 5 hexane/1 ethyl acetate followed by elution of the product with 3 hexane/1 ethyl acetate to provide 8.2 g (57%) of the monoalkylated product, 3- [1- [2- [1- (tertbutyldiphenylsilyloxymethyl) -3- ( tertbutyldimethyl -silyloxy) ropyloxy] ethyl] -3-indolyl] -4- (3-indolyl) -1-methyl-lH-pyrrole-2, 5-dione. MS. NMR.

X-8S51A FOR - SO - To a..methanol (450 mL! solution of the tert- butyldimethyisilyi ether, 3- [1- [2- C 1- (tertbutyicLLphenyisiiyioxy- methyi) -3- (tar butyid-unethyl-silyioxy) propyloxy] ethyl] -3-indciyi ]· -4- (3-indolyi) -i-methyl-iK-pyrroie-2, 5-dione (3.2 g, 9.9 aanol) under nitrogen at 5 °C was added p- . toluenesuifonic add, monohydrata (0.16 g, .035 eq) . After 2 hours, TLC (50% ethyl acetate/hexane) showed the reaction to be nearly complete. The reaction was quenc d with solid sodium • bicarbonate (0.14 g) - The methanol was removed in vacuo. The resulting residue was dissolved in ethyl acetate, washed with O-IN sodium hydroxide, brine (2X) , dried (KgSC^) , filtered and concentrated in vacuo to give a red foam. This material was absorbed on silica and placed on. a silica pad. Elution with 2 "hexane/1 ethyl acetate removed residual starting material .. ' followed by elution with 1 hexane/1 ethyl acetate and 1 hexane/2 ethyl acetate to provide 6.4 g (91%) of the alcohol, 3-[ l-[2-fl- ( ertbutyldiphenylsilylcxymethyl) -3- 'hydroxy. ropyloxy] ethyl] τ3- iadolyl] -4- [3-indolyl] -i-caethyl-lH-pyrrole-2, 5-dione. MS. NMR.

To an anhydrous ether (500 niL) solution of the alcohol, 3-[l- [2- [1- (tertbutyidiphenyisilyioxyiaethyl) -3- hydroxy' ropyloxy] ethyl] -3-indolyll -4- [3-indolyl] -l-∑ae.thyl-lK-pyrrole-2, 5-dione (6.36 g, 8.9 rnmol) under nitrogen, at 5aC was added triethyl- amine (1.9 mL, 1.5 eq) and ' ethanesulfonyl chloride (1.0 toL, 1.5 eq] . After 3 hours, additional triethylaniine (1.25 ruL, 1.0 eq) ..·. and methanesulfonyl chloride (0.7 mL, 1.0 eq) were added. After'.1. hour, the reaction was shown to be complete by TLC (50% ethyl acetate/hexane) . The reaction: was diluted with ether (250 ml.) , ' washed with water, 0.L HCl and. brine (2X) . The ' ether was dried (MgS04) , filtered, and concentrated n vacuo to provide 7.0 g of mesylate, 3-[l-[2-ri-(tertbutyldiphenylsilyIoxymethyl) -3- (methanesuifonyioxy). ropyloxy] ethyl] -3-indolyl! -4- [3-indolyi] -l-methyl-lH-pyrrola-2, 5-dione . MS..

To an acetone (200 mL) solution of the mesylate, 3-[i- 2- Cl- (tertbutyidiphenylsiiyloxymethyl) -3- 2-8351A FOE - €1 - (met anesulfcr.yioxy) propyloxy] e h^ -i-methyl-iH-pyrro-ie-2,5-dione, (7.0 c, 3.S tanal) under nitrogen was added sodium iodide (13.3 g, 10 eq) and sodium bicarbonate (75 nig,. 0.1 eq) . The mijctrore was stirred at 50CC far.13 hours. T e reaction was can entraned in vacuo, and the residua was dissolved in. ether and washed with 10-% scdiur-i : sulfi e solution. ' .The layers were separated, . and the ether pcricn wash d with 10% sodium sulfite solution, water, brine (22), dried, ≠τ*π concentrated in vacuo. .The residue was passed through a silica pad by eiuting with 1 hexane/1 ethyl acetate- and.1 he_ane/2 ethyl acetate.to provide 1.5 g of the iodide, 3- [I- [2- [1- (tartbutyidiphenylsilyiosyiiethyi; -3-ioccprcoyioxy] ethyl] -3-indoiylJ -4- [3-indalyl] -l-methyi-iH-pyrraIe-2, 5-diona . as a. red solid (quantitative yield., for. the- two steps) HS-. 2ffl£&. o a duae hy-lforaamid (1 L) suspension of cesium, carbonate (12.0 g, 4 eq) under nitrogen was added the iodide, 3- [1- [2- [1- ( ert utyldiphsnylsilyloxymethyl} -3-icdcprcpyioxy] ethyil-3-ir.dolyl]-4-[3-indclyi]-l-siethyi-lH-pyrroie-2, 5-dione (7.5 g, S -2 mmol) , dissolved in comethylformaniid (25 mL) via syringe pump over 65 hours. Three hours after the addition was ·complete, the reaction was ■ . concentrated in vacuo. The residue was dissolved in ethyl acetate (700 mL) , washed with water- (2 X 3G0 mL) , and- the aqueous -laye .backwashed with ethyl acetate (2 X .200 mL), The . combined ethyl acetate" portions were washed with brine (2 X 200-mL} , dried CMgSO^ , filtered and concentrated in vacuo to provide a purple residue.. The material was absorbed onto silica and applied to a flash column. Sl ad with -3 hexana/l ethyl acetate and then lhexane/1 ethyl acetate to give 5.2 g(82¾) of the macrocycie, 3, 4-[i, V C4-oxa-3-'(tertbutyldiphanyisiiylo-xy-methyl)hexano] -bis- (3-indoiyl) -l-methy MS. biMR.

A suspension of the itf-methyl maieimide, · 3, 4- [ L, 1' - [4-oxa-3- (tartbutyidiphanyisiiyLoxymethyl) hexanc] -his- (3-indoiyi) ] -1- -methyi-lH-pyrrcie-2,5-dione in 5M Ofi (150 mL) . and ethanoi (300 mL) was stirred at room temperature for 65 hours and then for one hour at SG^C. The reac ion was concentrated (150 mL) in.vacuo, the residue suspended, in water, 122092/4 X-S951A FOP. - 62 - cooled to 5 °C, and acidified (pH 3) with concentrated hydrochlor c acid. The red aqueous suspension was extracted with ethyl acetate (4 X 200 mL) , dried, and concentrated in vacuo to give 3.3 g of the crude anhydride alcohol, 3, 4- [I, 1' - (4-oxa-3- (hydxoxymethyi) hexar.o] -bis- ( 3- intiolyl) ] -furan-2 , 5-dione as a purple solid. H5.

To a dimethyiformamide (250 mL) solution of the anhydride 3, - [1, 1' - [4-oxa-3- (hydroxymethyi) hexar.o] -bis-(3- indoiyl) J -furan-2 , 5-dione, (3.3 c, 7.5 amcl) under nitrogen was added 1,1,1, 3, 3, 3 - heaamethyldisilaaane (32 mL, 2 eq) and methanol (3 mL, 10 eq) . The reaction was stirred at room temperature for IS hours and then heated at 60° for 2 hours. The dimethylforaamide was removed in vacuo, and the resulting residue was dissolved in acetonitrile (250 mL) . in HCl (50 mL) was added. The reaction was stirred for 15 minutes. The reaction was concentrated, partitioned between ethyl acetate (1 L) and water (250 mL) . The product was a solid that precipitated giving the alcohol maleimide, 3, 4-{ 1, 1' - [4-oxa-3-(hydroxymethyi) hexano] -bis- (3-indolyl) ] -iH-pyrrole-2, 5-dione, 0.92 g (28%) of product. A small amount (50 mg) was absorbed on silica and applied to a flash column, eiuted with dichloromethane, 5% acetonitrile/dichloromethane and then 10% acetonitoile/dic±loromethane to give 38 mg of analytically pure material. The ethyl acetate was concentrated and chroma. ographed to give an additional 8% of the crude product.

MS. lH NMR (d€-DKSO): δΐ.96 (IE, a}; 2.09 (1H, m) ; 3.31 ( LK, a); 3.40 (IE, m); 3.51 (1H, m) ; 3.62 (IE. m) ; 3.89 (IE, m) ; 4.13 (3E, a); 4.35 (12, a), 4.68 (IE, t, J = 2 Hz) 7.11 (2E, a) ; 7.19 (2E, m); 7.44 (IE, s) 7.46 (IE, d, J = 9 Hz}; 7.51 (IS, s) 7.53 (1H, d. .7 = 9 Hz); 7.79 (1H, d, J = 8 Ez); 7.83 (L£, d, J » 3 Ez) ; 10.91 (IE, s) . 122092/2 X-8951A FOR - 63 - Example 4 3, 4- [1, 1' - [4- (aminomethyl) -3-oxahexano] -bis- (3-indolyl) ] -lH-pyrrole- 2,5-dione trifluoroacetate salt To an anhydrous tetrahydrofuran (15 mL) solution of the alcohol, 3, 4- [1, 1' - [4- (hydroxymethyl) -3-oxahexano] -bis- (3- indolyl) ] -lH-pyrrole-2, 5-dione, (155 mg, 0.35 mmol) under nitrogen was added 2, 4, 6-collidine (280 μΐ>, 3 eg) . The solution was cooled to -78°C and treated with trifluoromethanesulfonic anhydride (118 μχ., 2 eg) . After 1,5 hours at -78°C, a large excess of concentrated ammonium hydroxide (2 mL) was added. After 10 minutes, the reaction was warmed to -42°C with a dry ice/acetonitrile bath and then stirred for 18 hours while allowing to warm to room temperature.

The reaction was concentrated in vacuo. The resulting residue was dissolved in ethyl acetate (400 mL) , washed with water, brine, dried, and concentrated in vacuo to provide the crude primary amine. The amine was absorbed on silica and applied to a flash column which was sequentially eluted with 1 ethyl acetate/1 hexane, ethyl acetate, ethyl acetate/5% methanol and finally 50 ethyl acetate/45 acetonitrile/4 methanol/2 isopropylamine to elute the amine, 3, - [1, 1' - [4- (aminomethyl) -3-oxahexano] -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (38 mg) .

Starting alcohol (104 mg, 67%) was also recovered. The product was further purified using reverse phase size exclusion chromatography by eluting with 85 acetonitrile/15 (0.01% TFA/water) . The collected fractions azeotroped with ethyl acetate to give 23 mg(12%) of a powder as the TFA salt.

MS. 122092/2 X-8951A -64- *H NMR (de-D SO) : 81.99 (1H, m) ; 2.08 (1H, m) ; 2.82 (1H, m) ; 3.18 (1H, ra) ; 3.57 (2H, m) ; 3.75 (1H, m) ; 4.13 (2H, m) ; 4.29 (1H, m) ; 4.44 (1H, m) ; 7.09 (2H, t, J » 7 Hz); 7.18 (2H, t, J = 7 Hz); 7.47 (4H, m) ; 7.70 (3H, bs); 7.78 (2H, m) In a analogous manner the S-enantiomer, 4 s as the HC1 salt, and the R-enantiomer, 4r as the HC1 salt, were prepared.

Example 5 3, 4-[l, 1 ,-[3-oxa-4-(dimethylaminomethyl) hexano]bis-(3-indolyl) ] - lH-pyrrole-2,5'-dione HC1 Salt To an anhydrous dichloromethane (140 mL) suspension of the alcohol, 3, 4- [1, 11 - [3-oxa- (4-hydroxymethyl) hexanojbis- (3-indolyl) -lH-pyrrole-2, 5-dione (472 mg, 1.07 mmol) under nitrogen was added pyridine (260 \xL, 3 eq) and methanesulfonic anhydride (242 mg,1.3 eq) . After 4 hours, the reaction was diluted with dichloromethane, washed with 0.1N HCl (2X) and filtered to remove starting material (54 mg) . The dichloromethane portion was washed with brine (2X), dried, and concentrated to give the crude mesylate, as a purple solid. The material was absorbed on silica and applied to a flash column which was sequentially eluted with dichloromethane, 5% acetonitrile/dichloromethane and 10% acetonitrile/ dichloromethane to provide 288 mg (52% yield) of the mesylate, 3, - [1, 11 -[3-oxa-4 (methanesulfonyloxymethyl) hexano] bis- (3- indolyl) ] -IH-pyrrole-2 /5-dione. MS. NMR.

To a tetrahydrofuran (20 mL) solution of the mesylate, 3, 4- [1, 1' - [3-oxa-4- (methanesulfonyloxymethyl) hexano] bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione, (304 mg, 0,59 mmol) was added a 8.9 M solution of dimethylamine in tetrahydrofuran (7 mL, 100 eq) . After heating X-8S51A -65- (55 °C) for 24 hours in a sealed tube, the reaction was diluted with ethyl acetate (200 mL) , washed with brine (2X) , cried, and concentrated to provide the crude derivative as a solid. The material was absorbed on silica and applied to a flash cciuffia. that was sequentially eluted with 3 ethyl acetate/ 1 kexace, ethyl acetate and 2% isoercpylaaiine/ethyi acetate to give the d ?,τηethyl?πir.¾ derivative 133 mg (70% yield} which was 90% pure by E?LC. The dimethylamine derivative, 3, 4- [1, 1' - [3-oxa (dimethylamiaomethyl) hexano] bis- (3-indolyl) ] -lH-pyrroie-2, 5- dione,was purified to greater than 95% as the trifluoroacetate salt using reverse phase size exclusion E3LC by eiuting with 85 acetonitrile/15 (0.01%TFA/water) .

The trifluoroacetate salt of 3, 4-[l, l'-[3-oxa-4- (dimethylaminomethyi) hexano] bis- (3-iadolyl) J -lE-pyrroie-2, 5-dione was converted to the KCl salt by suspending the salt in ethyl acetate aad washing gently with 0.1N NaOK(5 X 50 mL) . The ethyl acetate portion was washed with brine (2X) , dried, and concentrated to provide the free base, 3, 4-(l, 1' - f 3-oxa-4- (dime hylaminomethyl) hexano] bis- (3-iadolyl) ] -LK-pyrrole-2 , 5-dione. To an anhydrous methanol (50 mL) suspension of the free base, 3, 4-(l, 1' -(3-oxa-4- (dimethylaiainomethyl) hexano ] bis- (3-indolyl) ] -lK-pyrrole-2, 5-dione was added 1M HCi in anhydrous ether (13 mL, 50eq) . The ether was evaporated, and the residue was dried under vacuum to give 143 mg (52% yield) of 3, 4- (1,1'- (3-oxa-4- (dimethylamincraethyl) hexano]bis- (3-indolyl) 1 -lK-pyrrole -2, 5-dione hydrochloride salt as a red solid. MS.

!H NMR (de-DMSO) : 52.03 (1H, m) ; 2.26 (IE, m) ; 2.53 (6H, t, uT = 5 Hz); 3.24 (1H, m) ; 3.23, (1H, a, after D20 shake); 3.64 (IH, m); 3.77 (2H, m) ; 4.07 - 4.33 <4E, m) ; 7.03 (2H, ) ; 7.17 (2fl, m) ; 7.43 (3H, m> ; 7.52 (IH, d, J" - 8 Ez) ; 7.79 (2H, a); 10.33 (IH, bs) ; 10.52 (IE, s) X-8951A -57- Exampie 5r (R) -3, 4- [I, I' - [3-oxa-4- (di-nethylaiainomethyl) hexano] -bis- (3- indolyl) ] -lH-pyrrole-2 , 5-dione Hydrochloride Sale The R enantiomer was prepared in an identical manner as the (S) enantiomer, except using the (R)-4-ter - butyldiphenylsilyioxy-3- (2-iodoethoxy) -1-iodobutane as a starting material, MS. NMR.

Example 6 3, 4-[l, 1' -C3-oxa-5- (hydroxymethyl") heptano ] bis- (3 -indolyl) ] -1H- pyrrole-2, 5-dione A dry DMF (100 mL) solution of bis (mesylate), i-{2- (methylsulfonyloxy) -ethoxy) -2- ( (methylsulfonyloxy) ethyl) -3- (tert-butyldiphenyisilyloxy) -propane, (7.40g, 13.30 ittmoi) and 3,4-bis-(3-indolyl) -l-methyi-lH-pyrrole-2 , 5-dibne (4.43 g, 13.30 . ππαοΐ) was added over .16 hours to a stirred suspension of CS2CO3 (25.4 g, 78 mmol) in DMF (400 mL). at 50 aC . After 8 hours, the reaction was concentrated under reduced pressure at 80 °C to give a residue. The residue was diluted with ethyl acetate (200 mL) , washed with water (50 mL) . The organic layer was separated, and the aqueous layer was extracted with ethyl - acetate (50 mL x 3). The combined organic portion was dried,' and concentrated to a residue. The residue was eluted through, a column of silica gel with 25% ethyl acetate i hexane followed by 5% methanol in CH2CI2 to give three predominant products: The silyl ether macrocycle product,' 3, -[l, 1' - C 3-oxs-5- art-butyidiphenylsilyioxymethyl) heptano] bis- (3-indolyI) ] -1-methyl-lH-pyrrole-2, 5-dione (2.35 g) X-8951A -68- 122092/2 MS: Calculated for C44H45N304Si : Mol. mass : 707.31, found 708, Rf = 0.84 (50% ethyl acetate in hexane) , the desilylatad alcohol macrocycle product . (600 mg) . MS.

To a stirred EtOH (500 mL) solution of N-methyl macrocycle 3, 4- [1, 1' - [3-oxa-5- (tertbutyldiphenylsilyloxy-methyl) heptano] -bis- (3-indolyl) ] -l-methyl-lH-pyrrole-2, 5-dione , (1.65 g, 2.33 mmol) was added 5N KOH (100 mL) . After 12 hours at 50°C, the reaction mixture was cooled- to room temperature and concentrated under reduced pressure to a residue. The residue was acidified with concentrated HCl to pH 1 and extracted with ethyl acetate (200 mL x 5) . The combined organic phase was dried, concentrated under reduced pressure, and was eluted through a short silica column with 5% methanol in dichloromethane . Evaporation of the eluting solvent gave a residue containing the anhydride, 3, - [1, 1' - [3-oxa-5- (tert-butyldiphenylsilyloxymethyl) heptano] -bis- (3-indolyl) ] -furan-2, 5-dione that was used ir. the next reaction.

To a dry DMF (250mL) solution of anhydride, 3, - [1, 1' - [3-oxa-5- (tert-butyldiphenylsilyloxymethyl) heptano] -bis- (3-indolyl) ] -furan-2, 5-dione, (600 mg, 1.3 mmol) was added HMDS (2.1 g, 13 mmol) followed by methanol (209 mg, 6.5 mmol). After 48 hours, the reaction was concentrated, and the residue dissolved in ethyl acetate (100 mL) , washed with 1 N aq HCl (25 mL) , water (25 mL) and brine (25 mL) respectively. The resulting organic phase was then dried and concentrated to give a residue. The residue was eluted through a column of silica gel with methanol/CH2Cl2 (0% to 5% methanol) . Evaporation of the eluting solvent gave the imide, 3, - [1, 1' - [3-oxa-5- (hydroxymeth l) heptano bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione, as a .solid (300 mg, 50% yield) . MS. 1H NMR (CDCI3) δ 9.65 (s, 1H), 7.79 (t, J = 7.65 Hz), 7.61 (s, 1H), 7.54 (s, 1H), 7.46-7.40 (m, 2H) , 7.24-7.08 ( , 2H) , 7.07-7.02 (m, 2H) . 4.43-4.33 (m, 2H) , 4.30-4.21 (m, 1H) , 4.14-4.06 (m, 1H), 3.64 (t, J = 4.64 Hz), 3.58-3.38 (m, 5H) , 3.71 (t, J = 8.64 Hz, 1H), 1.89-1.85 (m, 1H) Χ-8951» Examnie / 3, 4- [1, 11 - j-ox-5,-5- (l-pyrrolidinomethyl) hegta o] bis- (3- indolyl} ] -lE-oyrrola-2,.5-dione hydrochloride salt To a dry CH2 12 (50 JnL) solution of imide alcohol, 3, 4- [1,1 ' - [3-oxa-5-(hydroxymeth.yi ) heptanol bi5~(3-indoiyi) ] -1Ξ-pyrrcie-2, 5-dione (140 mg, 0.30 mmol} containing pyridine (120· mg, 1.5 nnaoi) was added methane sulfonic anhydride (IQS.j-g, 0.61 mmol) under a N atmosphere. After 12 hours, . the reaction was quenched with water (25 mL), diluted with CH2C12 (50 mL) , washed with 0.2 N ECl (20 mL x 2), aq sodium bicarbonate (20 mL) , water (20 mL) , brine (20 mL) , dried, and concentrated to a residua... The residue was eluted through a short silica gel column with 5% methanol . in dichloromethane and evaporation of the . eluting solvent gave the mesylate, 3, 4- [1, 1 ' - [3-oxa-5~ (niethanesulfonyloxymethyi) heptano]"bis- (3-indolyl) ] -IK-pyrrole- . 2,5-dione that was used in the next reaction.

To a sealed tube THF (20 mL) solution of mesylate , 3, 4- CI, 1 ' - [3-oxa-5- (me hanesul onyloxym. th. i) heptano] bis- (3-' indolyl) ]-lH-pyrroie-2,5-dione, (157 mg, 0.29 mmpi} was added pyrrolidine (203 mg, 2.90 mmol) . After heating (50 °C) for' 12 hours, the reaction was cooled to room temperature, concentrated under reduced pressure, dissolved in CH2 12 (50 mL) , washed with water (20 mL x 2Y, brine (20 mL) , dried, and concentrated under reduced pressure to. a residue. The residue was eluted through a. short silica gel column with 5% methanol' in dichidromethaha and evaporation of the eiutiag solvent. gave the pyrrolidine, 3,4-[1,1'- [3-oxa-5- (1-pyrrolidinomethyi) heptano] bis- (3-indolyi) ] -1H-pyrroie-2, 5-dione, MS.: calcul ted for C31K32N4O cl mass: 5QS.52, found 503, f = 0.14 (5% methanol in dichlorcmethane, X-8951A -70- 122092/2 trace triethylamine) . The pyrrolidine was further purified by reverse phase gel permeation chromatography to give the pyrrole macrocycle as the trifluoroacetic acid salt (55 mg, 37% yield) .

The trifluoroacetic acid salt of the pyrrole was converted to the hydrochloride titled compound by extracting a 1 N NaOH (5 mL) slurry of the trifluoroacetic acid salt (55 mg) with ethyl acetate (25 mL) / methanol (2 mL) , the extract was dried, and concentrated to a residue. The residue was slurried in ether/methanol (10:1) and a HC1 solution of ether was added.

After 30 minutes, the slurry was concentrated and dried in vacuo to give the title compound (48 mg, 88% yield) . MS.

½ NMR: δ 10.98 (s, 1H) , 7.90 (s, 1H) , 7.82 ( s, 1H) , 7.70-7.62 (m, 3H) , 7.56-7.50 (m, 1H) , 7.24-7.02 (m, 4H) , 4.50-4.20 (m, 4H) , 3.76-3.42 (m, 4H) , 2.82-2.44 (m, 4H) , 2.26-2.24 (m, 1H) , 1.82-1.60 (m, 6H) , 1.26-1.02 (m, 2H) .

Example 8 3, 4- [1, 1 ' - [3-oxa-5- (N, N-dimethylaminomethyl) heptano]bis- (3- indolyl) ] -lH-pyrrole-2, 5-dione hydrochloride salt The titled tertiary amine was prepared by displacement of the mesylate with dimethylamine (58 mg, 75% yield) . MS.

XH (CDC13) δ 10.93 (s, 1H), 7.84 (s, 1H) , 7.77 (s, 1H) , 7.69-7.64 (m, 3H) , 7.47 (d, J = 7.97 Hz, 1H) , 7.13-7.02 (m, 4H) , 4.40-4.11 (m, 4H) , 3.73-3.20 (m, 4H) , 2.50 (s, 3H) , 2.33 (s, 1H) , 2.13- 1.96 (m, 2H) , 1.86-1.70 (m, 1H) , 1.21-1.10 (m, 2H) The following compounds were prepared in a manner analogous to the Examples described herein and further illustrate the compounds of the invention. In the following examples, the structure of the compound was confirmed by NMR, MS, and/or elemental analysis. During the synthesis, R is a protected hydroxy, preferably a silyl ether preferably tert- X-8951A -71- 122092/2 butyldiphenylsilyloxy (TBDPS) . The silyl ether may be converted to a leaving group and substituted to produce the following examples : Example n nl n2 9 3 2 0 -NH2 10 3 2 0 -NH2 · HC1 11 3 2 0 -N(CH3)2 · HC1 13 3 2 0 -NHCH2C6H5 ■ HC1 14 3 2 0 -NHCOCH3 15 3 2 0 -NHSO2C6H5 16 3 2 0 -NHC(0)OCH2C6H5 17s S-enantiomer 2 2 0 -NHCH3 · HC1 18s S-enantiomer 2 2 0 -NHCH2C6H5 · HC1 18r R-enantiomer 19r R-enantiomer 2Or R-enantiomer 20s S-enantiomer 21r R-enantiomer 2 2 -NHSO2C6H5 21s S-enantiomer 22 -NHCH2 (py idyl) · HC1 —N N-CH3 23s S-enantiomer HC1 —N O 24s S-enantiomer 2 2 0 · HCl 25r R-enantiomer 2 2 0 -NHC(0)OCH2C6H5 3, 4- [1, 1' - (methano [2, 6]pyridomethano) -bis- (3-indolyl) ] -IK -pyrrole-2 , 5-dione .

Following the same procedure as described in Example 1, 3, 4-bis- (3-indolyl) furan-2, 5-dione (287 mg,0.88 mmol) in 5 mL DMF was treated with sodium hydride (60 percent in oil, 88 mg, 2.19 mmol) for 1.5 hours, then diluted to 11 mL with DMF, and treated with 2, 6-bis-bromomethyl pyridine (245 mg 0.93 mmol) . After stirring at 50eC overnight, the reaction mixture was worked up (EtOAc) and filtered through a short plug of silica (50 percent EtOAc in hexanes) . 3, 4- [1, 1' - (methano [2, 6] pyridomethano) -bis- (3- indolyl) ] furan-2, 5-dione (142 mg, 37 percent) was obtained as a dark red solid, which showed essentially a single spot on the TLC analysis and was used directly in the next step without further purification. 3, 4- [1, 1' - (methano [2, 6Jpyridomethano) -bis- (3-indolyl ) ] furan -2, 5-dione (140 mg, 0.32 mmol) in 2 mL of DMF was treated with a mixture of 1,1,1,3,3,3- hexamethyldisilazane (0.72 mL, 3.2 mmol) and CH3OH (0.063 mL,1.6 mmol) to give, after workup and purification by radial chromatography on silica gel, 42 mg of the titled, 3, - [1, 1' - (methano [2, 6] pyridomethano) -bis- (3-indolyl) ] -1H- pyrrole-2, 5- dione, as a burgundy solid. This material was homogeneous by TLC (Rf = 0.35, 3 percent CH3OH in CHC13) .

X-8951A -73- 122092/2 Example 27 3, 4- [1, 1' - (ethanoxy [1, 2] benzenomethano) -bis- (3-indoly-.) ] - lH-pyrrole-2, 5-dione hydrochloride A dry DMF (45 mL) solution of dibromide, 2-(2'- Bromoethoxy) -benzylbromide, (2.0 g, 6.8 rnmol) and3, 4-bis- (3- indolyl)-l-methyl-lH-pyrrole-2, 5-dione (2.3 g, 6.8 rnmol) was added via syringe pump over 20 hours to a suspension of CS2CO3 (8.9 g, 27 rnmol) in dry DMF (550 mL) with vigorous stirring at 55°C under N2. After an additional 2 hours, the reaction mixture was concentrated in vacuo, the residue dissolved in CH2CI2 washed with IN HC1,, brine, dried, and concentrated in vacuo to give a violet oil. The oil was passed through a plug of silica eluting with 1:1 hexanes/ethyl acetate. The eluant was reduced to yield the macrocycle, 3, 4- [1, 1' - (ethanoxy [1, 2] benzenomethano) -bis- (3-indolyl) ] -l-methyl-lH-pyrrole-2, 5-dione 2.76 g (71% yield) as a magenta solid. ecrystallization from isopropanol/methylene chloride gave analytically pure material. MS: M = 473; observed 473, FD, CHCI3) , EA: Calculated (observed): C 76.09 (75.86); H 4.90 (4.93), N 8.87 (8.79).

To an ethanol (100 mL) suspension of macrocycle, 3,4- [1,1'- (ethanoxy [1, 2] benzenomethano) -bis- (3-indolyl) ] -1-rr.ethyl-lH-pyrrole-2, 5-dione (710 mg, 15 rnmol) containing THF (20 mL) was added 5N KOH (80 mL) . The reaction was heated (55°C) for 70 hours with stirring, cooled to room temperature, and the ethanol removed in vacuo. The concentrate was acidified to pH 1 with 5N HC1 (325 mL) , extracted with ethyl acetate, washed with brine (2x), dried, and concentrated to give the anhydride, 3,4-[1, 1' - (ethanoxy [1, 2 ] benzenomethano) -bis- (3-indolyl) ] -furan-2, 5 -dione 700 mg (quantitative conversion) as a residue.

X-8951A -74- 122092/2 To a dry DMF (500 mL) solution of the anhydride, 3,4- [1,1'- (ethanox [1, 2] benzenomethano) -bis- (3-indolyl) ] -furan-2, 5 - dione (760 g, 17 mmol), was added a solution of methanol (0.34 mL, 8.3 mmol) and 1, 1, 1, 3, 3, 3-hexamethyldisilazane (3.5 mL, 17 mmol) . After heating (55°C) 22 hours the reaction was concentrated in vacua, diluted with ethyl acetate, washed with 0.1N HCl. The combined organic layer was dried, and concentrated to a violet residue. The residue was applied to a short plug of silica and eluted with CH2Cl2/hexane (gradient 0 - 100% CH2CI2) · Evaporation of the eluting solvent gave the NH maleimide, 3,4- [1, 1' -(ethanox [1, 2] benzenomethano) -bis- (3-indolyl) ] -IK - pyrrole-2, 5-dione 483 mg (70% yield) as a' purple solid. The title compound was crystallized from CH2Cl2/hexane. MS.

XH NMR: (DMS0-d6) δ 4.29 (2H, bs) 4.59 (2H, bs) ; 5.23 (2H, bs); 6.90-6.99 (2H) , 7.01-7.18 (3H) , 7.20-7.27 (2H) , 7.59-7.68 (2H), 7.71-7.80 (5H);10.92 (H, s) .

Example 28 3, 4- [ (1, 1 ' -hexano) -bis- (3-indolyl) 1 -lH-pyrrole-2, 5-dione.

To a solution of 3, -bis- (3-indolyl) -1-methyl-lH-pyrrole-2, 5-dione (499 mg, 1.46 mmol) in 10 mL of DMF under N2 was added sodium hydride (60 percent in oil, 146 mg, 3.65 mmol) in portions of 30 minutes. The resultant green solution was stirred 1 hour. The mixture was diluted with 10 mL of DMF and then treated dropwise with 1, 6-dibromohexane (0.24 mL, 1.57 mmol) . The reaction mixture was stirred 30 minutes at room temperature and then heated at 45°C for 16 hours. The cooled mixture was poured into dilute aqueous NH4CI (125 mL) and X-SSS1A -75- extracced with EtGAc {3 x 40 mL) . The combined organic extracts were washed with water and dried (2igSQ4) . After removal of the solvent in vacczo, the residue was purified by flash chromatography on silica gel eiuting with CE∑Ciz-hexane5.. 1:1 to 3:1 (gradient eiution) to afford compound 3, 4- [ (1, 1' -hexano) -bis- (3-indoiyl) ] -i-methyi-lE- pyrrole-2, 5-dione (137 ag, 22 percent) as a purple solid, . Pt. > 320°C.

A mixture containing 3, 4-Γ (1, 1T -hexano) -bis- (3-indolyi)] -l-methyl-lH-pyrrole-2, 5-dioGe(137. mg, 322 mmol), ethanoi ■ (15 mL) , 5 N. KOE (5 mL) and ΓΞΖ (2 mL) was stirred 4 hours at room temperature. At that time · LC analysis showed. 'the-' 'starting material to be consumed. The mixture was' diluted -with water (.15 mL), and concentrated on the rotary evaporator. The mixture was cooled, acidified, to. pE.l with.3 N ECl and extracted with CH?Ci2 ( 3 x 10 mL) . The combined organic extracts were washed ..well with water, dried over anhydrous teqSO* and concentrated. The purple solid obtained (116 mg) was found by NMR analysis to be a 4:1 mixture of the desired anhydride and starting material.

This material was used directly in the next step without further purification.

In the same manner as that described in Example 1, a ' solution of 3, 4-[ ( 1 , 1 ' -hexano) -bis- (3-indolyi) -furan-2,·5-dione (108 mg, 0.263 mmol) in DMF (1.5 mL under N2 was treated with .a . mixture of 1, 1, 1, 3, 3, 3-hexamethyldisilazane (0.53 mL, 2.62 mmol) and C¾QE (0.05 mL, 1.31 mmol) overnight. After worku (EtQAc) , the crude product was subjected to flash chromatography on, silica gel (CH2Clz-EtCAc, 10:1 - 5 :i,. gradient elution) to afford two colored fractions. The first colored fraction to eiuate contained, the.3, 4- [ (1, 1 ' -hexano j -his- (3-indoiyl} - L-methyl-lE-pyrroie-2, 5-dibne impurity carried from the previous reactions. The second colored, fraction contained the desired product, 3, 4- [.(1, 1 ' -hexano ) -bis- (3-indoiyi)j-lH-pyrroie-2, S-dione (56 mg) . M. Ft. > 320*C. MS Calculated for (0.3 ¾G) : C, 76.26; H, 5.66, N, 10.26.

Found : C, 75.21; E, 5.-65; N, 10.05.

X-8951A -76- 122092/2 Example 29 3, 4- C 1, 1 '- (3' '-((benzylcarbonato) methyl) hexano) -bis- (3- indolyl) ] -lH-pyrrole-2, 5-dione.

To a 0° C dichloromethane solution of 3, 4- [ (1, 1 ' - (3 · ' - (hydroxy methyl) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (24 mg, 0.054 mmol) was added diisopropylethylamine (10.6 mg, 0.081 mmol) followed by benzyl chloroformate (13.8 mg, 0.081 mmol) . After 72 hours, the reaction mixture was quenched with 2.5 N sodium bicarbonate; the organic layer removed; and the aqueous layer extracted with dichloromethane. The combined organic layers were combined, washed with brine, dried over magnesium sulfate, filtered and concentrated to give an oil that was purified by reverse phase HPLC (5% acetonitrile in wate with 0.1% TFA gradient to 100% acetonitrile on CI 8 column) to give 6 mg of the title compound. MS. (±) -3, -[ 1,1'- (3"- (benzyloxymethyl) hexano) -bis- (3-indolyl) ] -1H- pyrrole-2, 5-dione Following the same procedure as described in the previous examples, 3, 4-bis- (.3 '-indolyl) -l-methyl-lH-pyrrole-2, 5-dione (400 mg, 1.17 mmol) in 8 mL DMF was treated with sodium hydride (60 percent in oil, 117 mg, 2.93 mmol) followed by X-8951A -77- 122092/2 treatment with (±) -3-benzyloxymethyl-l, 6-dibromohexane in 7 ml of DMF. After heating at 50°C overnight, the crude product after workup was purified by flash chromatography on silica gel eluting with CH2Cl2-hexanes, 1:1 - 2:1 (gradient-elution) to give pure (±)-3,4-[ 1/ l1- (3"- (benzyloxymethyl) hexano) -bis- (3- indolyl)] -l-methyl-lH-pyrrole-2, 5-dione(149 mg 23 percent) as a violet solid.

A mixture containing (±)-3,4-[ l,l'-(3"-(benzyloxymethyl) hexano) -bis- (3-indolyl)]-l-methyl-iH-pyrrole-2, 5- dione (141 mg, 0.259 mmol), ethanol 15 mL and 5 N KOH (5 mL) was stirred at room temperature for 3 hours at which time TLC analysis showed the starting material to be consumed. After acidification and extraction with CH2CI2, the crude product (101 mg) showed two spots on TLC analysis (CH2CI2) corresponding to starting material and desired anhydride (±)-3,4-[ l,l'-(3"-(benzyloxymethyl ) hexano) -bis- (3-indolyl)]-furan-2, 5-dione . NMR analysis indicated roughly a 4:1 mixture of anhydride and starting material respectively. This material was used directly in the next "step without further purification. (±)-3,4-[ 1, 1'- (3 "-(Benzyloxymethyl) hexano) -bis- (3- indolyl)]-furan-2, 5-dione (98 mg, 0.180 mmol) in 1 mL DMF was treated with a mixture of 1, 1, 1, 3, 3, 3-hexamethyldisilazane (0.41 mL, 1.80 mmol) and CH3OH (0.036 mL, 0.90 mmol) at 25°C overnight. The mixture was worked-up (EtOAc) and flash chromatographed on silica gel eluting with CH2CI2 CH2Cl2~EtOAc 10:1 (gradient elution) to give 30 mg of purified (±) 3, 4- [1, 1 '- (3"- (benzyloxy methyl) hexano) -bis- (3-indolyl)]-lH-pyrrole-2, 5-dione. M. Pt. 171°-173°C. MS -8951A -78- Exa-roie 31 3, 4- [ 1/ 1 ' - (3 hydroxymethyl) hexano) -bis- (3-incolyl) ] -IE- pyrrole-2, 5-dlone A mixture, containing 3, 4- bis- (3-indolyl) -I- methyl -pyrroie-2, 5-dione (3.41 g, 10.0. mmol} and 3-tsrt-butylciiphenylsilyloxymethyiane-l,.6-d^i^ (5..64 g, 11.0 -caoi) in 50 mL of DKF ,.was added with a syringe- pump over. 30 hours to a well-stirred solution of. cesium carbonate (11.2 g, 34.3 mmol) in DMF (350- mL) at 55° C under Na.. After the addition was complete, the reaction mixture was heated at this temperature an .additional 16 hours. The cooled mixture was . poured into 1.2 L of water containing.20 mL of 3N HC1 and extracted with three 300 mL portions of CE2C12. The combined organic extracts were washed with water' and brine, then dried (MgSC). and concentrated. The residue was passed through a 3" x 3" column of .silica gel eluting . with- HCi^. The crude product thus, obtained, was .purified by flash chromatography on silica gel (CHCla). o afford 2.87 g (41%) of 3,4-[ 1/ 1 ' - (3' ' -(fcert-butyldiphenyisiiyloxyme hy1) hexano) -bis- (3-inc0lyl) 3 - -methyl-LE pyrrole-2, 5-dione as a purple solid, M. pt. 220 - 224° C. E?¾S calculated fox' CM+ ] : 692.3307. Found: 632.3299. .

A mixture containing 3,4-[ 1, 1 ' - ( 3 ' ' -fcert-butyldiphenyisiiyioxymethyl) hexano) -bis- (3-indolyi) ] -i-methyl-ΙΞ pyrrole-2, 5-dione (1.55 g,. 2.22 xamoi) , 4N KOH (100 mL) , , THF (10 mL) and 95% EtOE (200 mL) was heated at .30°C for 16 hours.

After removal .of most of the EtOE on the rotary evaporator, the mixture was acidified to pH 1 with S EC! and' extracted with CE2CI2 (3 x 75 mL) . The combined organic extracts were washed with water and brine and dried ever anhydrous Na23Q4. After removal of the solvents in vacua, the residue was dissolved into X-3SSIA -79- a w-ί ^ τ τηι'ιτη of 5% met anol i CHCI3 and loaded onto a 3" x 3™ column of silica gal. Elation wit CEC13 followed by 10 % methanol in CEC13 gave two fractions; evaporation of t e se ond fraction provided €76 nig. .(69%) of anhydride-alcohol as a purple solid which was homogeneous by TLC (Rf = 0.5, 10% 'methanol in' CHC13) .. This material was used directly in the next ste without further purification. · To a solution of . the above anhydride (510 mg, 1.15 mmol) in ΕΗΞ* (11 mL) was added a premixad solution containing 1,1,1,3,3,3,- hexasethyidi-sil z ne (3.14 mL, 23 mmol) and CEjOE . (0.45 mL, 11.5 mmol). The- resultant mixture was heated at 5C°C for 24 hours under N2- The cooled reaction mixture was poured into.100 mL of water. The .precipitated product was washed with, water and dried overnight, to give -409 ag of .3,4- C i/ ,-(3''r (hydroxymethyi) hexano) -bis- (3-indoiyi) ] -iZ-pyrrole-2, 5-diona as .a . reddish-purple solid. This material was found to be 93% pure b EPLC (reverse-phase) analysis and was contaminated with an unidentified compound.of similar Rf. ERMS calculated . for ...

: C27E2SN303 : 439.1896. Sound: 439.1911.

Example 31r (R) -3,4- C 1, 1' - (3 ' '- (hydroxymethyl) hexano) -bis- (3-indolyl)-} -IS- ' pyrrole-2>.5-dione Following the same procedure described above -for the preparation of Example 31, (R)-3,4-[ 1 , 1 ' - (31 ' -hydroxyme hyl) - hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-diona was prepared in 25% overall yield from the dibromide, . (R) - 3- (tert-butyldiphen- yisiiyloxymethyi) -1, 6-dibromohexane by dial2yiation of3, 4-bis- (3- indolyi) ] -l-methyi-lE-pyrrole-2, S-dione, hydrolysis, and 1-Ξ- . pyrroie-2, 5-dione formation. M. pt. > 300°C. . .

E NM (300 MHz, DMSO-d6)51 · 05. - 2.25 (m, 7Ξ) , 4.04 - 4.45 (m, 6Ξ). (,m, 8H) , 7.08 -.7.88 (m, 10 E) .

X-3S51A -30- Sxp.Tn ls 3 Is (S) -3, -£ 1, 1'- (3' '-(hydroxy st yl.) -hexano) -bis- ( 3-indolyl) I-1H- pyrrole-2/ 5-dione Following tiia same procedure described above for the preparation- of Example 31, (3}-3,4-[ l,l'-(3"-Cnydroxymethyi .] -hexano) -bis- (3 -incclyi) ] -lH-pyrrois-2, 5- . diona was prepared (4.5 g) ir. 39% overall yield from dib-ciaida, (S) -3- (tert-b tyldiphenyisilyloxyTaetriyl} -1, by diaixylation of 3, 4- bis- (3-indolyi) -l-mathyl-IH-pyrroie-2, 5-dicn hydrolysis/ and iE-pyrroie-2, 5-diona formation. MS.' Έ,Μ (c , DKSO) 5 1.05-1.15 (2≤), 1.23-1.24 (IS), 1.50-1.52 (IE), 1.71 ( IE) / 1.94 (IE) , 2.07- .12 ( IE) , 4.05-4.4 (a, 6Ξ} / . 7.09-7.21 Examples 32 and 33 3, 4-[ l,l'-(3' '-{amiJiomethyl)hexano)-bi5-(3-incolyl) I -lE-pyrrole- 2 , 5-dione Example 32 as the ΈΓΑ salt .

Example 33 as" tha'HCl Salt To a stirred solution of the anhydride alcohol 3,4- l,l'-(3' '-(hydroxymethyl)hexano) -bis- (3-indolyi) ] -furan-2, 5-dione (0.18 g, 0.41 mmoi) in anhydrous dichloromethane (10 mL) , under nitrogen was added trie hylamine (0.10 g, "l.QS mmci) , and methanasuifonyichloride (0.11 g, Q.S3 Emol) . The resulting solution was stirred 30 minutes at room temperature. The solvent was removed in vacuo. The residue was dissolved . in 10 aL anhydrous dimei^yifaraamide, followed by the addition of sodium azide .( 0".26 g, 4.1 mmoi).. The reaction mixture was X-8951A -81- 122092/2 heated for 1.5 hours at 50° C under nitrogen. The cooled reaction mixture was partitioned between 0.2 N HCl and ethyl acetate. The combined organic extract was dried over magnesium sulfate, filtered, and evaporated to provide 185 mg of the azide which was used directly in the next reaction.

The crude azide was dissolved in dimethylformamide (3 mL) , under nitrogen, and 1, 1, 1, 3, 3, 3-hexamethyldisilazane (1.25 g, 7.75 mmol) and methanol {0.12 g, 3.87 mmol) were added. The reaction was heated at 50° C. After 12 hours, the reaction was cooled, diluted with ethyl acetate, washed with water, hydrochloric acid 2 N. The aqueous washes were back extracted with ethyl acetate (3x50 mL) . The combined organic layers were dried over magnesium sulfate, filtered, and evaporated to provide the azide-intide, 3,4-[ 1, 1" -(3' '-(azidomethyl) hexano) - bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (175 mg) as a purple colored solid. The product was chromatographed on a Rainin Dynamex R -60 Ci8 column ( 21.4x250 mm) using a linear gradient from 805 A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 minutes at 15 mL/min. to obtain purified 3,4-[ 1, 1 (3' '-(azidomethyl) hexano) -bis- (3-indolyl) ] - lH-pyrrole-2, 5-dione in 57% overall yield. MS. NMR.

To a solution of the azide 3,4-[ 1,1' -(31 1-(azidomethyl) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (0.1 g 0.21 mmol), in ethyl acetate (15 mL) and ethanol (5 mL) was added Lindlar's catalyst (0.1 g) . The reaction mixture was stirred under hydrogen (1 ATM) at room temperature. After 12 hours, the catalyst was removed by filtration and the filtrate was concentrated in vacuo. Purification by preparative reverse phase HPLC on a Rainin Dynamax R- 60 C18 (21.4x250 mm) using a linear gradient 80% A (0.1 % TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 minutes at 15 mL/min., provided the primary amine as the TFA salt, 3,4-[ Ι,Ι'-Ο11-(aminomethyl) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione trifluoroacetic acid salt, as a solid (80 mg) in 63% yield. MS. lH NMR (d6 acetone) δ 0.77-0.78 (m, 1H) , 1.0-1,1 (m, IK), 1.27-1.34 (m, 1H), 1.43 (m, 1H) , 1.52-1.56 (m, 4H) , 1.60-1.1.68 (m, 1H) , X-8951A -82- 122092/2 1.90-1.94 (m, 1H) , 3.17-3.21 (m, 1H) , 3.35, 3.38 (m, 1H) , 3.64- 3.67 (m,lH), 3.75-3.82 (m, 2H) , 6.61-6.72 (m, 4H) , 6.824 (d, J= 16 Hz, 2H), 6.936 (t, J= 8.31 Hz, 2H) , 7.397 (t, J=7.83 Hz, 2H) , 9.3 (s, 1H) . 13C NMR (d6 acetone) δ 26.0, 28.0, 32.1, 35.4, 40.8, 41.0, 41.1, 45.1, 45.8, 50.9, 105.1, 105.2, 110.8, 111.0, 121.24, 121.29, 122.7, 122.9, 123.0, 128.4, 128.6, 131.5, 132.0, 134.0, 134.1, 136.8, 137.1, 172.6, 172.7, 192.5 The HCl salt is prepared as described on page 70.

Example 34 3, 4- [ 1, 1 (31 '-((N-benzylamino) methyl) hexano) -bis- (3-indolyl) ] - lH-pyrrole-2, 5-dione trifluoroacetate salt To a stirred solution of the primary amine, 3,4- [ 1,1' -(3* aminomethyl) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (40 mg, 0.05 mraol) , in anhydrous THF (2 mL) under nitrogen was added benzaldehyde (9.39 mg, 0.08 mmol) . After 30 minutes, sodium triacetoxy borohydride (18.75 mg, 0.08 mmol) was added. After stirring 1 hour, the reaction mixture was diluted with water and extracted with ethyl acetate (3x25 mL) . The combined organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo. Purification by reverse phase HPLC on a Rainin Dynamax R- 60 C18 column (21.4x250 mm) using a linear gradient from 80% A ( 0.1% TFA and 5% acetonitrile in water) to 100% B (pure acetonitrile) over 60 minutes at 15 mL/min., provided two different fractions of mono benzyl compound, 3,4-[ 1, 1 ' -( 3 ' * -((N-benzylamino) methyl) hexano) -bis- (3-indolyl) ] -1H-pyrrole-2, 5-dione (16 mg) in 66% yield, and the dibenzylamino compound, 3,4-[ 1,1'- (3' '-((Ν,Ν-dibenzylamino) methyl) hexano) -bis-(3-indolyl) ] -lH-pyrrole-2, 5-dione, (7 mg) in 20% yield. MS.

X-8951A -83- 122092/2 1H NMR (d6 acetone) δ 1.1-1.3 (m, 1H) , 1.5-1.6 (m, 1 H) , 1.71- 1.77 (m, 1H), 1.93-2.10 (m, 3H) , 2.5 (m , 1H) , 3.1-3.2 (m, 1H) , 3.37-3.41 (m,lH), 4.13 (t, J= 5.1Hz, 2H),4.28 (t, J= 5.1 Hz, 2H), 4.36 (d, J= 3.6 Hz, 2H) , 7.13-7.24 (m, 4H) , 7.33 (d, J= 25 Hz, 2H), 7.39-7.51 (m, 7H) , 7.89-7.96 (m, 2H) , 9.76 (s, 1H) . 13C NMR (d6 acetone) δ 25.6, 27.3, 32.1, 32.9, 44.7, 45.4, 50.1,52.2, 105.0, 105.2, 110.8, 111.1, 121.2, 121.3, 122.8, 122.9, 123.1, 128.5, 129.8, 130.3, 131.2, 131.3, 132.0, 132.4, 133.7, 134.0, 136.8, 137.0, 172.5, 172.6 Example 35 3, 4- [ 1,1'- (3' '-((N^-dibenzylaminojmethyDhexano) -bis- (3- indolyl) ] -lH-pyrrole-2, 5-dione trifluoroacetate salt 3,4-[ 1,1*- (3* '-{(Ν,Ν-dibenzylamino) methyl) hexano) -bis-(3-indolyl) ] -lH-pyrrole-2, 5-dione was prepared in a manner analogous to Example 34. MS.

¾ NMR (d6 acetone) δ 0.2-0.3 (m, 1H) , 0.6-0.9 (m, 4H) , 1.2-1.3 (m, 1H) 1.50 (d, J= 5.4 Hz, 2H) , 2.27 (m, 1H) , 3.3-3.8 (m, 8H) , 6.6-6.9 (m, 18H), 7.35 (dd, J= 7.5 Hz, J=24.9 Hz, 2H) , 9.1 (s, 1H) .

X-8951A -84- 122092/2 Example 36r (R)-3,4-[ Ι,Ι'-Ο' '-(l-pyrrolidino methyl) hexano) -bis- (3- indolyl) ] -lH-pyrrole-2, 5-dione hydrochloride salt A stirred mixture of mesylate, (R)-3,4-[ Ι,Ι'-Ο'»- ((methanesulfonyloxy)methyl) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione, (202 mg) and pyrrolidine (1.5 mL) in THF (15 mL) was heated at 50°C until TLC indicated the starting material be consumed (16 hours) . EtQAc (30 mL) was added. The organic phase was washed with 10 mL portions of 5% NaHC03, water and brine. Concentration afforded a deep-red residue which was subjected to preparative HPLC (Waters reverse-phase, 0.1% TFA and 5% CH3CN in water- 100% CH3CN gradient) to give pure (R)-3,4-[ l,l'-(3' '-(l-pyrrolidinomethyl)hexano) -bis- (3-indolyl) ]-lH-pyrrole-2, 5-dione as its TFA salt. Conversion to HCl-salt in the same manner gave (R)-3,4-[ 1, 1'- (3' '-(l-pyrrolidinomethyl) -hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione hydrochloride salt (42 mg) as a light red solid. M. pt. 220° C (dec.) . HRMS calculated for C31H33N4O2 [M+l] : 493.2604. Found : 493.2605.

X-8951A -85- 122092/2 Example 37 3,4-[ l,l'-(3' '-(methoxymethyl)hexano) -bis- (3-indolyl) ]-1Η- pyrrole-2, 5-dione A solution of 3,4-[ 1, 1 ' - (3 » · -(tert-butyldiphenylsilyloxymethyUhexano) -bis- (3-indolyl) ] -1-methyl-lH-pyrrole-2, 5-dione (1.25 g, 1.81 mmol) in THF (20 mL) was treated with a solution of tetra-n-butylammonium fluoride in THF (1M, 2.0 mL, 2.0 mmol). The mixture was stirred for 1 hour at 25 °C. The reaction mixture was quenched with IN HC1 (5 mL) and diluted with EtOAc (75 mL) . After washing with water and brine, the organic layer was dried (MgS04) and concentrated. The residue was "purified by flash chromatography on silica gel eluting with 3-5% methanol in THF/hexanes (1 :1) to afford alcohol, 3,4-[ 1, 1 (3 ' 1 -(hydroxymethyl) hexano) -bis- (3-indolyl) ] -1-methyl-lH- pyrrole-2, 5-dione, 509 mg (62 %) as a purple solid. This material was used directly in the next step.

To a stirred solution containing the above alcohol (285 mg, 0.63 mmol) and 47% aqueous tetrafluoroboric acid (170 mg, 0.95 mmol) in CH2CI2 (6 mL) at 0 C was added dropwise a solution of trimethylsilyldiazomethane (Aldrich, 2.0 M hexanes, 0.47 mL, 0.95 mmol) over 5 minutes. The resultant mixture was stirred at 0° C for 2 hours then at 25° C for 4 hours. TLC analysis of the reaction mixture indicated a large amount of unreacted starting material. The mixture was cooled and an equivalent additional amount of tetrafluoroboric acid trimethylsilyldiazomethane was added. The mixture was stirred 2 hours at o" C then 6 hours at 25° C and diluted with CH2CI2 (20 mL) and washed with 2N HCl (10 mL) and water (10 mL) . The organic layer was dried (MgS04), concentrated. The residue was loaded onto a 3" x 3" silica gel column and eluted with CH2CI2 to give methyl ether, 3,4-[ 1,1'- X-8951A -86- 122092/2 (3 ' ' -(methoxymethyl) hexano) -bis- (3-indolyl) ] -1-methyl-lH-pyrrole- 2,5-dione, 114 mg, (39%) as a reddish-purple solid, M.?t. 234 - 236°C. MR.

HRMS calculated for C29H29N3O3 : 467.2208. Found : 467.2210.

A mixture of 3,4-[ 1/ 1 '- (3 ' · -(nethoxymethyl) hexano) - bis- (3-indolyl) 3 -l-methyl-lH-pyrrole-2 5-dione (110 mg, 0.243 mmol), and SN KOH (8 mL) in 15 mL of EtOH containing 1 mL of THF was heated at 90°C for 24 hours. After removal of most of the ethanol under reduced pressure, the mixture was acidified to pH 1 with 6N HC1 and extracted with CH2CI2 (3 x 15 mL) . The combined organic extracts were washed with dilute aqueous NaHC03 and water and dried over anhydrous MgS04. After removal of the solvents in vacuo, the crude product was loaded onto a 2" x 2" column of silica gel and eluted with CH2CI2 to give anhydride which was used directly in the next reaction.

To a solution of the above anhydride (76 mg, 0.17 mmol) in DMF (1.5 mL) was added a solution of 1,1,1,3,3,3-hexamethyldisilazane (0.75 mL, 3.34 mmol) and CH3OH (0.07 mL, 1.67 mmol) that had been pre ixed for 5 minutes The reaction mixture was stirred 1 hour at 25° C then heated at 50° C for 20 hours whereupon TLC analysis showed the reaction to be complete. The cooled reaction mixture was worked up (EtOAc) as previously described. The crude product was purified by flash chromatography on silica gel (CH2CI2 - 4% EtOAc in CH2CI2/ gradient elution) to afford 42 mg (55%) of 3,4- [ Ι,Ι'-Ο*1-(methoxymethyl ) -hexano) -bis- (3 -indolyl) ] -lH-pyrrole-2 , 5-dione as a dark red solid. Recrystallization from acetone-water gave 28 mg. of analytically pure 3,4-[ l,l'-(3' '-faethoxymethyl) -hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione as reddish-violet solid, M.Pt. 272 - 274 °C .

Analytical calculated for C28H27N3O3 (0.1 H2O) : C, 73.86; H, 6.02; N, 9.23.

Found : C, 73.51; H, 5.92; N, 8.99.

X-8951A -87- 122092/2 Example 38 3,4-[ 1,1'-(3' '-(acetoxy methyl) hexano) -bis- (3-indolyl) 1 -1H- pyrrole-2, 5-dione Acetic anhydride (0.064 itiL, 0.68 mmol) was added to a stirred mixture of the anhydride, 3,4-[ l,l'-(3'!- (hydroxymethyl) -hexano) -bis- (3-indolyl) ] -furan-2, 5-dione (1.49 mg, 0.34 mmol), 4-dimethylaminopyridine (27 mg, 0.22 mmol), pyridine (0.75 mL) and THF (1.5 mL) . The reaction mixture was stirred at 25° C under N2 for 16 hours. The mixture was diluted with EtOAc (20 mL) and washed with 2N HC1 (2 x 10 mL) and water (2 x 10 mL) and dried over anhydrous MgS04. After evaporation of the solvents under reduced pressure the crude product was purified by chromatography on a short column of silica gel eluting with CH2CI2 to give the 0-acetate anhydride, 3,4-[ 1,1'- (3' '- (acetoxymethyl) hexano) -bis- (3-indolyl) ] -furan-2, 5-dione, 111 mg, (68%) as a purple solid, M.Pt. 252 - 254°C.

To a stirred solution of the 0-acetate anhydride, 3,4-[ 1, 1 ' - (3' '- (acetoxymethyl) hexano) -bis- (3-indolyl) ] -furan-2, 5-dione, (103 mg, 0.22 mmol) in DMF (2 mL) was added a solution containing 1, 1, 1, 3, 3, 3-hexamethyldisilazane (0.48 mL, 2.2 mmol) and CH3OH (0.043 mL, 1.1 mmol) which had been premixed for 5 minutes. The reaction mixture was worked up (EtOAc) as previously described and the crude product was purified by flash chromatography on silica gel (gradient elution: CH2CI2 - 5% EtOAc in CH2CI2) to give the 0-acetyl maleimide, 3,4-[ 1,1'-(3 ' '- (acetoxy methyl) hexano) -bis- (3-indolyl) ] -lH-pyrrol -2, 5-dione 74 mg (72%) as a deep red solid which was homogeneous by X-8951A -88- TLC (CH2CI2) · Recrystallization from acetone-water provided the titled compound as a red solid. M.Pt. 250 - 252° C.

Analytical calculated for C29H27N3O4 (0.1 H2O) : C, 72.06; Η,· 5.67; N, 8.69.

Found: C, 71.72; H, 5.67; N, 8.29.

The following compounds were prepared in a manner analogous to the Examples described and further illustrate the compounds of the invention. In the following examples, the structure was confirmed by NMR, MS and/or elemental analysis.

Example 39 - HC(0)OCH2(C6H5) 40 -N(CH3)2 · HC1 X-8951A -89- 122092/2 Example 4Or (R)-3,4-[ 1,1'- (3' '-((N,N-dimethylamino)methyl)hexano)-bis-(3- indolyl) ]-lH-pyrrole-2, 5-dione hydrochloride salt Methanesulfonic anhydride (94 mg, 0.5 mmol} was added over 10 minutes to a stirred solution of chiral alcohol, (R) - 3,4-[ 1, l1- (3"- (hydroxy methyl) hexano) -bis- (3-indolyl) ] -1H-pyrrole-2, 5-dione (200 mg, 0.45 mmol), and pyridine (0.11 mL, 1.35 mmol) in CH2CI2 (5 mL) at 0° C. The reaction mixture was stirred at 25° C for 4 hours. CH2CI2 (20 mL) was added, and the mixture was washed with 10 mL portions of 3% HC1, water and brine and dried over anhydrous MgS04. Removal of the solvent in vacuo left the crude mesylate (205 mg) which was homogeneous by TLC (1% methanol in CHCI3. This material was carried on directly to the next step.

To a solution of the above mesylate (205 mg) in 10 mL of THF was added 40% aqueous dimethylamine (2 mL) and the reaction mixture was heated at 50° C for 36 hours. After removal of the THF under reduced pressure, CH2CI2 (20 mL) was added to the residue. The mixture was washed with 5% aqueous NaHC03, water and brine and dried over anhydrous MgS04.

Concentration afforded crude (R)-3,4-[ 1,1'- (31 '-((N,N-dimethy1amino) methyl) he ano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione (158 mg) as a red solid that was purified by preparative HPLC (Waters reverse-phase, 0.1 % TFA and 5% CH3CN in water-100% CH3CN gradient) to give the amine-TFA salt which was dissolved in CH2CI2 and converted to the free base with dilute aqueous OH. After drying the organic phase over MgS04 (15 X-3951A -90- minutes) , the solvent was evaporated and. the free amine (60 ng) was dissolved into 1:1 aethanoi/TEF (S mL) , .ceolad. to C C under N2 and slowly acidified to pH 4-5 ..{external damp pE paper} with anhydrous IN EC! in ether . The precipitated salt was filtered and washed with dry ether under a N2 bianxat then dried in a vacuum desiccation over CaSQ4 overnight. The dime hyla ine-ECl salt, (R)-3,4-[ l^l'-iS^ iN/N-d seth iami o-imet yi )-hexano-} -bis- (3 -indoiyl) ] -lE-pyrrole-2, 5-dicne hydrochloride salt (43- mg) was obtained as a light red solid, M.=t. 230° C (dec.) . MS. ,lE H (300 ¾Ez, acatone-d«) δ 0. - 3.5 (m, 7E) ., 3.20· - 3.42 On, 8Ξ), 4.05 - 4.18 ( , 4S) , 7.02 - 7.80 (m, 10S) , 10.94(s, IE).

Example 40s (S) -3, 4-[ 1, 1 '- (3* ' -((N/ N-dimethylaznino) aethyl ) hexano) -bis- (3- indolyl) 3 -lH-pyxrole-2, 5-dione hydrochloride salt . Following the same procedure described above -for the preparation of Example 40r, (S)-3,4-[ 1, 1 ' - (3' * -((N,N-dimethyiamina) methyl) hexano) -bis- (3-indolyl) ] -iS-pyrrola-2 , 5-dione hydrochloride salt was prepared (90 mg) in 27¾ overall yield from the alcohol/ (S)-3,4-[ i, 1 ' - (3 ' ' -(hydroxymeth l) -hexano). -bis- (3-indolyl) ] -lS-pyrrcle-2, 5-dione.. by formation of the mesylate and displacement with dimeth lamine ■ MS . 1NMR (d« DMSO) 5. 0.92 (a large,! Ξ) , " 1.35 (s large, .1 E) , 1.60 (s large, 2 E) , 1.85 (s large, IE) , 2.37-2.42 (m, 2E) , 2.91-3.05 (m, 2Ξ) , 4.13 (s large/ 2Ξ) , 4.23 (s large/ '.2Ξ) , 7.11-7.23 (a, 4S), 7.34 (d, J= 20 Ez, 2S) , 7.50 (dd, J= 3.1 Ez, J= 12.6 Ez, 2E), 7.79 (d, j- 3 Ez, 2Ξ) , 9.92 (a large/ IS) , 10.93 (s, IS) X-8951A -91-.

ExamDla 41 3, 4-Γ I, 1'- (3 ' '-{(lH-imldazol-l-yl) methyl) hexano i -bis- (3- ... iadolyl) ] -ig-pyrrole-2, 5-dione . He hanesulfcnyl chloride. (Q.Q25_mL, 0.32 mmol) was added dropwise to a stirred solution containing 3, - f i,i'-(3''- (hydroxy methyl) hex?.no) -bis- (3-indolyl) ] -LK-pyrrol -2, 5-<±Lone (100 mg, 0.23 mmol) and triethylimine' (0.05 mL, 0.36 mmci) in- dry CECI3 at 25° C under N2 - After stirring for 20 minutes/ the ■ reaction mixture was diluted .with CHCI3 (15 mL) , washed with water, ,brine., dried filtered and concentrated. The red residue was purified by chromatography on a short column of silica gel eluting with CHCl3 followed by 10% EtOAc.in CSCl3...to give 3/ - ' £ 1, 1 ' - (3 ' '-(methaneSTilfonyloxymethyl ) -hexano) -bis- (3 — · indolyl) ] -lE-pyrrole-2, 5-dione , 53- mg, as. a red solid, which was homogeneous by TLC 5% (EtOAc in CE2CI2) · To a stirred solution of 3,4-( ·1,1'-(3' '- ((aethanesulfonyloxy) methyl) hexano).-bis- (3-indolyl) ] -LH-pyrroia- 2, -dione (49 mg, 0.0S5 mmol) in DMT (.0.75 mL) under U2 was added dropwise a solution of the sodium salt of .imidazole in D F (prepared by adding 60% NaE (8.7 mg, 0.22 mmol) to a solution of imidazole (16 mg, 0.24 mmol) in DMF (0.5 mL)) . The reaction .\ mixture was stirred 15 minutes at 25° C then heated at 5G°C for 30- minutes. The reaction mixture was dilu ad with 25 mL of CE2C12 containing 3% methanol. The mixture was washed with 10 ■mL portions of water and brine and dried over anhydrous Na23G . After evaporation of the solvents under reduced pressure, the crude product was ■ loaded onto a 3" x 3" column of silica gel X-8951A -92- 122092/2 eluting with CH2CI2 followed by 5% methanol in CH2CI2 containing 1% triethylamine to afford 3,4-[ 1, l1 - (3' '-((lH-imidazol-l-yl) methyl ) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione, 21.5 mg (46%) as a red solid. This material was subjected to reverse-phase HPLC (gradient elution, 5% CH3CN in water containing 0.1% TFA - CH3CN) to provide analytically pure 3,4-[ 1, 1 ' - (3 ' ' -((1H-imidazol-l-yl) methyl) hexano) -bis- (3-indolyl) ] -lH-pyrrole-2, 5-dione {12.4 mg) as a red solid, M.Pt. 261 -266°C. NMR. HRMS calculated for C30H27N5O2 [M+l] : 490.2244. Found : 490.2242.

The following compounds were prepared in a manner analogous to the Examples described herein and further illustrate the compounds of the invention. In the following examples, the structure of the compound was confirmed by NMR, MS, and/or elemental analysis.

Example n nl Q R 42 3 3 CH H 43 2 2 CH H 44 3 4 CH H 45 3 3 CH NH2 46 3 3 CH NHCOCH3 47 3 3 CH NHCH2C6H5 48 3 3 C (-OCH2CH2O 49 3 3 C =0 3, 4- [ ,1/ 1' - ί 4-thiaheptu.no) -bis- ( 3-indolylΠ -lH-pyrrole-2, 5- dione To a 0aC stirred anhydrous CH2Ci2 (1.0 L) solution of N- (3-acetoxypropyl) -indole (102 g, 0.47 mole) was added oxalyl chloride (43.04 mL, 0.494 mala , 1.05 eq.) dropwise . After 15 minutes, the ice bath was removed. The reaction mixture was . allowed to warm to ambient temperature with stirring for three hours. The volatiles were removed in vacuo to. yield a magenta solid, which was redissolved in dry CH2CI2 (1.0 1) under ¾2 · With vigorous stirring, -cert butoxycarbonyl-indole-3-acetic acid (129.25 g, 0.47 mole}, was added followed rapidly b triethylamine (130.6 mL, 0.94 mole, 2 eq.) . After 15 hours, the reaction was concentrated and purified by flash column' chromatography, eluting with 3:1 hexane/ethyl acetate. The major colored fraction was concentrated _to give the anhydride. (101 g, 40% yield) 3- [1- (3-acetoxypropyl) -3-indolyl] -4- (l-{cert-butoxycarbony¾-3-indolyl]-furan-2, 5-dione as a red crystallin solid. MS ..To the' BOC protected anhydride (7. g, 14 mmol) was added trifluoroacetic acid (27 inL, 35Q mmol) containing e.thanethiol (1 mL, 14 mmol) with stirring.. After one hour, the reaction mixture was partitioned between CH2CI2. and saturated , NafiC03. . The organic layer was washed with brine, dried over N 2S04, a d filtered. The filtrate was concentrated to give the crude deblocked anhydride as a red semi-solid. The residue was applied to a short pad of silica, washed with hexane and then CE2CL2. The colored band was eluted from, the silica wich ethyl acetate and dried isi vacuo to give the purified deblocked anhydride, 3- [1- (3- cetoxypropyl) -3-indolyl] -4- (3-indolyl) -curan-2,5-dione (5.7 g, 95% yield) as red solid. MS X-8951A -94- 122092/2 To a stirred anhydrous DMF (125 mL) solution of the deblocked anhydride (3.0 g, 7 mmol) was added NaH (420 mg, 10.5 mmol, 60% in mineral oil) at room temperature. A color change from bright orange to violet was immediately observed. After 30 minutes, 3 equivalents of 3-bromopropyl acetate was added rapidly. The reaction was heated to 75°C, and gradually returned to an orange color. After 6 hours, the DMF was removed in vacuo. The residue was applied to a flash silica chroiaatography column eluting with 3:2 hexane/ethyl acetate. The major red band was collected, and the solvent removed to give the alkylated anhydride, 3, 4'-bis-[l- (3-acetoxypropyl) -3- indolyl] -furan-2, 5-dione (1.32 g, 36%) as a red solid. MS 3, -bis [1- (3-acetoxypropyl) -3-indolyl] -furan-2, 5-dione (1.32 g, 2.52 mmol) was suspended in absolute ethanol (125 mL) with stirring and treated with 5N OH (125 mL) . After stirring for 16 hours, the reaction mixture was concentrated to 126 mL. The residue was acidified (5N HCl) slowly, until a red solid precipitated. The precipitate was filtered and dried in a. vacuum oven -at 60°C, producing the alcohol anhydride 1,1 g (99%) as a red powder.

The alcohol anhydride, (1.1 g, 2.47 mmol) was dissolved in anhydrous DMF (30 mL) under a N2 atmosphere with stirring. A premixed solution of 1, 1, 1, 3, 3, 3-hexamethyl-disilazane (5.22 mL, 24.7 mmol, 10 eq.) and methanol (0,50 mL, 12.4 mmol, 5 eq.) was added. The reaction was stirred for 16 hours at ambient temperature. The DMF was removed in vacuo. To this residue was added acetone (100 mL) and excess CsF (ca. 500 mg) . After stirring 4 hours, the reaction was concentrated. The residue was partitioned between ethyl acetate and water. The organic layer was washed with IN HCl (5x) , brine (2x) , dried over Na2S04 and filtered. The filtrate was concentrated to give the bisindolylmaleimide 3, -bis[l-(3-hydroxypropyl) -3-indolyl] -lif-pyrrole-2, 5-dione 1.0 g (91% yield) as a red powder. Total yield was 90% over two steps. MS 3, 4-bis [1- (3-hydroxypropyl) -3-indolyl] -lH-pyrrole-2, 5-diona (1.0 g, 2.25 mmol) was dissolved in anhydrous CH2CI2 (250 mL) at ambient temperature under N2. CBr4 (2.09 g, 6.3 mmol, X-8951A -95- 122092/2 2.8 eq) and triphenylphosphine (2.83 g, 10.8 mmol, 4.8 eq.) were added together to the reaction vessel. The mixture was stirred for 16 hours. The crude reaction mixture was concentrated and purified by silica gel flash column chromatography, eluting with 7:3 hexane/ethyl acetate. The desired product eluted as one major red band. Removal of the solvents from this fraction gave the dibromo compound, 3,4-bis [1- (3-bromopropyl) -3-indolyl] -lH-pyrrole-2, 5-dione 876 mg (68% yield) as a red powder.

The dibromo compound (47.8 mg, 0.08 mmol) was dissolved in acetone at ambient temperature with stirring. An excess of sodium sulfide nonahydrate (229 mg, 0.95 mmol, 11.3 eq.) was added. The heterogeneous mixture was stirred overnight. The acetone was then removed in vacuo. The residue was partitioned between water and CH2CI2. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give 35.5 mg (94% yield) of the titled product as an red-orange solid. 3,4-[ l,l'-(3"-(hydroxy methyl) pentano) -bis- (3-indolyl) 3-1 H. - pyrrole-2, 5-dione A dry DMF (35 mL) solution of 1, 5-diiodo-3- ( tert-butyldiphenylsilyloxymethyD-pentane (7.3g,12 mmol)and3, 4-bis-(3-indolyl)]-l H -pyrrole-2, 5-dione (4.21g, 12 mmol) was added via syringe pump over 48 hours to a suspension of CS2CO3 (16.06g, 49.3 mmol) in dry DMF (1 L) with vigorous stirring at 55°C under Ν2· After an additional 2 hours, the reaction mixture was concentrated in vacuo, the residue dissolved in CH2CI2, washed with IN HC1, brine, dried, and concentrated in vacuo to give a X-8951A -96- vioiet oil. The oil was passed through a plug of silica eluting with 4:1 hexanes/ethyl acetate. The eluant was reduced to yield the macrocycle, '3,4-1 1 1'- (3' '- ( tert-butyldiphenylsilyloxy-metiiyl) pentano) -bis- (3-indolyl} ]-l methyl. -IE -pyrrole-2, 5-dione, 4.5g (55% yield) as a magenta solid.

To an ethanoi (300 mL) suspension of 3,4-[ l,l'-(31'- (tert-butyldiphenyisilyloxymethyi) entano ) -bis- (3-indoiyl) methyi-lH-pyrrole-2, 5-dione (4.2g, 6.2 mmol) was added 5N QH (300 mL) . The reaction was refluxed (8S°C) for 43 hours with' stirring, cooled to room tem erat e/ and the ethanoi removed in vacuo. The concentrate was acidified to pE .1 with 5N ECi (325 . mL).,. extracted with ethyl acetate, washed with brine (2x) , dried, and concentrated to give' the anhydride, 3,4-[ l,l'-(3' '- (hydroxymethyl) pentano) -bis- (3-indolyl) ]-furan-2, 5-dione, 2.6 g (100% yield) as a residue.

To a dry DMF {500 mL) solution of the . anhydride, 3,4- E 1,1'- (3' '- (hydroxymethyl) pentano ) -bis- (3-indolyl) ] -furan-2, 5-dione (2.6 g," 6.2 mmol), was added a solution of methanol (1.25 mL, 31 mmol) and 1, 1, 1, 3, 3, 3-hexamethyldisilazane (13.1 mL, 62 mmol) . After heating (55°C) 36 hours the reaction was . concentrated in vacuo, diluted with ethyl acetate, washed with IN HC1 The acid wash contained some solids that were back extracted wit chloroform. The combined organic layer was dried, and concentrated to a violet residue. The residue was applied to a short plug of silica and eluted with 2-10% MeCN/CH2Cl2 · The fraction containing the major product is . concentrated in vacuo to yield the title alcohol, 3, 4- [ 1,1'- (3 '·- (hydroxymethyl) entano) -bis- (3-indolyl) ] -1 H -pyrrole-2, 5- · dione (650 mg (25%) ) . as a magenta solid. MS.

XE NMR: (DMSO-ds) δ 0.7 (m, 1H); 1.48 (m, 2H) ; 1.32 ( , 2H) ; 3.19 (dd, 2S) ; 4.16 (m, 4H) ; 4.4 (t, 1H) ; 7.05 (t, 2H) ; 7.16 (t, 2Ξ) ; 7.17 (s, 2H) ; 7. 6 (d, 2H) ; 7.65 (d, 2H) ; 10.96 (s, 1Ξ) ..

X-S951A -97- Examcle 3,4-[1,1'-(3' - (methanasulfonyloxymethyl) hexano) -bis- (3- indolyl)]-! H -pyrrola-2, 5-dione To a dry ¾Cl2 (80 mL) solution of 3,4-Cl,!'- (3' '-(hydroxyiaethyl)hea:ano--bis-{3-iiidolyl) ]-1 Ξ' -pyrrole-2, 5-dione (334120 j- (650 mg, 1.5 mmol) was added methanesulfonic anhydride (400 mg, 2.29 mmol) followed b excess pyridine (370 mL, 4.58 mmol) . After 16 hours at ambient temperature, the reaction mixture was applied directly to a short plug of silica and elutad" with 0-7% MeCN/Cii2Cl2 · The colored fraction was concentrated in vacuo to give the mesylate, 3, 4- [1, 1 ' - (3' '- (methanesulfonyloxymethyi) hexano) -bis- (3 -indolyl)]-! H -pyrrole-2, 5-dione 501 mg (67% yield) of a violet solid. MS. ·. , ' . '-lH NMR: (EMSO-de) δ 0.89 (m, IE); 1.61 (m, 2H),v 1.82 (m, 2H) ; . 2.99 (s, 3H) 4.02 (d, 2Ξ) ; 4.22 (m, 4H) ; 7.06^ (t,: 2H) 7.17 (t, 2H); 7.17 (s, 2H) ; 7.54 X-8951A -98- 122092/2 Example 53 3/4-[l,l'-(3"-(aminomethyl)pentano)-bis-(3-indolyl) ]-l H - pyrrole-2, 5 dione hydrochloride salt In a sealed tube reaction vessel containing a THF (20 itiL) solution of the mesylate 3, 4- [1, 1 ' - (3 ·' '- (methanesulfonyloxymethyl)pentano) -bis- (3-indolyl) ] -1 H - pyrrole-2, 5-dione (250 mg, 0.5 mmol) was added NH4OH (33% aq, 10 mL) , the reaction tube was sealed, and heated (60°C) . After 48 hours, the reaction mixture was cooled and eluted through a plug of silica gel with ethyl acetate followed by acetone. The acetone fraction was reduced in vacuo to give a reddish solid. A portion of this residue was purified using reverse phase gel filtration HPLC (85% MeC /water, 0.01% TFA) . The pure fractions were pooled and concentrated to a red solid. The solid was then partitioned between ethyl acetate/0. IN NaOH. The organic layer was concentrated to give the free base as a residue. The residue was dissolved in methanol (2 mL) and treated with HC1 (2 mL, 1.0 M in ether) for 1 hour. The reaction was concentrated in vacuo to yield the title compound 28.5 mg (13%) of a magenta solid which is >95% pure by HPLC analysis. MS.

XH NMR: (DMSO-de) 5 1.17 (m, 1H) ; 1.5-1.63 (m, 2H) 1.6-1.95 (m, 2H); 2.73 (m, 2H) ; 4.18 (m, 4H) ; 7.12 (t, 2H) ; 7.15 (s, 2H) ; 7.23 (t, 2H) 7.56 (d, 2H) 7.75 (d, 2H) ; 7.8 (br, 3H) ; 11.01 (s, 1H) .

X-8951A -99- 122092/2 Example 54 3,4-[ I, l'-(3"-((N/N- dimethylamino) methyl) pentano) -bis- (3- indolyl)]-! H -pyrrole-2, 5-dione hydrochloride The title compound was prepared as the hydrochloride salt by using dimethylamine (40% aq, 5 mL) to displace the mesylate 3, 4- [1, 1*-(3· ' -((methanesulfonyloxy) methyl) pentano) -bis- (3-indolyl) ] -1 H -pyrrole-2/ 5-dione (110 mg, 0.2 mmol) and subsequently transforming to the hydrochloride salt to produce the titled compound (28 mg, 26% yield) . MS.

H NMR: (DMSO-d6) 5 1.17 (m, 1H) ; 1.5-1.63 (m, 2H) ; 1.8-1.95 (m, 2H); 2.73 (m, 2H) ; 4.18 (m, 4H) ; 7.12 (t, 2H) ; 7.15 (s, 2H) ; 7.23 (t, 2ΗΓ; 7.56 (d, 2H) ; 7.75 (d, 2H) ; 7.8 (br, 3H) ; 11.01 (s, 1H) .

X-8951A FOR - 100 - The following compounds are prepared in an analogous manner and further illustrate the compounds of the invention: Example n nl W R 55 3 2 0 =CHCH2NHCH2C6H5 56 3 2 0 =CHCH20CQNH(C6H5) 57 3 2 0 =CHCH2NHCOCH3 58 3 2 0 \ =CHCH2NHS02C6H5 59 3 2 0 =CHCH2CH20H 60 3 2 0 =CHCH2CH2 H2 61 2 2 0 =CHCH20CONH(C6H5) 62 2 2 0 =CHCH2CH2OH 63 2 2 0 =CHCH2CH2 H2 64 2 2 CH =CHCH2 HCH3 65 2 2 CH =CHCH2NHS02C6H5 66 2 2 CH =CHCH2CH20H 67 2 2 CH =CHCH2CH2NHCH2C6H5 68 2 2 CH =CHCH2CH2NH2 69 2 2 CH =CHCH2CH2OC0 H (C6H5 ) 70 2 2 CH =CHCH2CH2 HS02CH3 71 2 2 CH =CHCH2CH2N(CH3)2 72 2 2 CH =CHCH2CH2NHCH3 73 2 2 CH =CHCH20CH2CH2NH2 .74 2 2 -OCH2- =CHCH2NH2 75 2 2 -OCH2- =CHCH2NHCH3 76 2 2 -OCH2- =CHCH2NHCH2C6H5 77 2 2 -OCH2- =CHCH20CONH(C6H5) 78 2 2 -OCH2- =CHCH2 HCOCH3 79 2 2 -OCH2- =CHCH2 HS02C6H5 80 2 2 -OCH2- =CHCH2CH2OH X-8951A FOR - 104 - 951A FOR - 10 - E2L. ¾L Rla ¾J2 Eli 88 H CH3 H H 89 OCH3 H H H 90 H CH3 CI H 91 H N02 H H 92 CF3 H H H 93 OH H I CH3 H 94 N(CH3)2 H H . H Example n nl Q R 95 3 3 S =0 96 3 3 S (=0)2 97 3 .3 0 -"- 98 3 3 CH -OH 99 3 3 CH OCO HC6H5 100 3 3 N H 101 3 3 N CH3 102 3 3 . CH HSO2C6H5 X-8951A FOR - 103 - 103 3 3 CH NHCH3 104 3 3 CH HCH2C6H5 108 3 3 CH CH2NH2 109 3 3 CH CH2NHCOCH3 110 3 3 CH CH2N ( CH3)2 111 3 3 CH CH2 HSO2C6H5 112 3 3 CH CH2 HCH2C6H5 113 2 2 C =0 As previously noted, the compounds of the present invention are potent, protein kinase inhibitors. The compounds are selective for protein kinase C over other kinases.

The ability of the compounds of the present invention to selectively inhibit protein kinase C was determined in the Calcium Calmodulin Dependent Protein Kinase Assay, Casein Protein Kinase II assay, cAMP-Dependent Protein Kinase Catalytic Subunit assay and the Protein-Tyrosine Kinase assay.

Calcium Calmodulin Dependent Protein Kinase Assay (CaM) The Calcium Calmodulin Dependent Protein Kinase Assay is described in the Journal of Neuroscience. 3_:818-831 (1983). The assay components are in a total volume of 250 μΐ·: 55 mM HEPES (4- (2-hydroxyethy1 ) -1-piperazine-ethanesulfonic acid), pH 7.5, 2.75 mM dithiothreitol , 2.2 mM EGTA (ethylenebis (oxyethylenenitrilo) tetraacetic acid, used in the blank buffer), 1.1 mM calcium chloride (Sigma, St. Louis, Missouri) (used in the control buffer), 10 mM magnesium chloride (Sigma, St. Louis, Missouri), 200 μg mL histone type HL (Worthington) , 10 μL DMSO or DMSO/inhibitor and 30 μΜ (gamma 32P) ATP (DuPont) . The reaction is initiated by the addition of calcium calmodulin dependent protein kinas (isolated from rat brain homogenate) , incubated at room temperature for 10 minutes and stopped by adding 0.5 mL ice cold trichloroacetic acid X-8951A FOR - 10. - (Amresco) followed by 100 [IL of 1 mg/mL bovine serum albumin (Sigma, St. Louis, Missouri). The precipitate is collected by vacuum filtration on glass fiber filters and quantified by counting in a beta scintillation counter.

Buffer components: Control buffer Blank buffer 200 mM HEPES pH 7.5 3125 jiL 625 |1L histone 1250 μΐ 250 μ-L 100 mM calcium 125 ^IL 100 mM EGTA 50 L DI water 2375 HL 450 |AL Assay components: 165 μΐ,. Buffer \ 25 μL calmodulin (250 μg/mL) 10 μL DMSO or DMSO/inhibitor 25 p kinase enzyme 25 \1L AT32P.

Casein Protein Kinase II Assay fCK-II) The Casein Protein Kinase II Assay is described in Neurochem. Res .. 13 : 829-836 (1988). The assay components are in a total volume of 250 μΐ^: 20 mM Tris-HCl, pH 7.5, 5 mM sodium fluoride, 50 mg/mL Casein (Sigma, St. Louis, Missouri), 10 mM magnesium chloride (Sigma, St. Louis, Missouri), 10 μL DMSO or DMSO/inhibitor and 30 μπι (gamma- 32P) ATP (DuPont) . Initiation of the reaction is performed by addition of casein protein kinase II (isolated from rat brain homogenate) , incubated at room temperature for 10 minutes and stopped by the addition of 0.5 mL ice cold Trichloroacetic acid (Amresco) followed by 100 μL of 1 mg/mL bovine serum albumin (Sigma, St. Louis, Missouri) . The precipitate is collected by vacuum filtration on glass fiber filters and quantified by counting in a beta scintillation counter.

X-8951A -105- Assay components in order of addition 175 ]i Buffer 10 μ-L oF DMSO or DMSO/inhibitor 25 ]i of AT32P in 300 uM magnesium chloride 40 pL of enzyme (undiluted) Buffer prepared as follows: (Final volume = 3.5 mL: amount of 20 assays) 500 i of each: 200 mM Tris-HCl pH 7.5 50 mM sodium fluoride 50 mg/mL Casein + 2 mL DI water Total Volume cAMP-Dependent Protein Kinase Catalytic Subunit Assay (ΡΚΆ) The Assay components are in a-total volume of 250 ]i : 20 mM HEPES (Sigma, St. Louis, Missouri) buffer pH 7.5, 200 pg/mL histone type HL (Worthington) , 10 mM magnesium chloride (Sigma, St. Louis, Missouri), 10 i DMSO or DMSO inhibitor and 30 uM (gamma- 32P) ATP¾ (DuPont) . The reaction is initiated by addition of bovine heart cAMP-dependent kinase . catalytic subunit (Sigma, St. Louis, Missouri), incubated to 30°C for 10 minutes and stopped by adding 0·.5 mL ice cold Trichloroacetic acid (Amresco) followed by 100 pL of 1 mg/mL bovine serum albumin (Sigma) . The precipitate is collected by vacuum filtration on glass fiber filters employing a TOMTEC™ and quantified by counting in a beta scintillation counter. This assay is done identical to the protein kinase C (PKC) enzyme assay except that no phospholipids or diacylglycerol are employed in the assay and the histone substrate · is specific for the cAMP-dependent catalytic subunit enzyme.

Protein Tyrosine Kinase Assay (src) The Assay components are the following: 10 mL Raytide 10 mL Kinase 4 mL DMSO or DMSO/inhibitor X-8951A FOR - 106 - 6 HL 200 mM HEPES pH 7.5 10 UL AT32P This assay is described by Onogene Science, Inc. Cat. #PK02 and PK03 (1990) .

Surprisingly, the compounds of the present invention are also isozyme-selective inhibitors, that is, the compounds selectively inhibit protein kinase C beta-1 and beta-2 isozymes. This isozyme selectivity was determined in the PKC Enzyme Assay.

PKC Enzvme Assay PKC enzymes = alpha, beta I, beta II, gamma, delta, epsilon, eta and zeta.

Assay components in a total volume of 250 μ!.· are as follows: Vesicles consisting of 120 ig/mL phosphatidylserine (Avanti Polar Lipids) and sufficient diacylgiycerol (Avanti Polar Lipids) to activate the enzyme to maximum activity in 20 mM HEPES buffer (Sigma, St. Louis, Missouri), pH 7.5, 940 μΜ calcium chloride (Sigma, St. Louis, Missouri) for assaying the alpha, beta-1, beta-2 and gamma enzyme only, 1 mM EGTA for all the enzymes, 10 mM magnesium chloride (Sigma, St. Louis, Missouri) and 30 μΜ (gamma-32P) ATP (DuPont) . For all the enzymes either histone type HL (Worthington) or myelin basic protein is used as substrate. The assay is started by addition of protein kinase C enzyme incubated at 30°C for 10 minutes and stopped by adding 0.5 mL of cold trichloroacetic acid (Amresco) followed by 100 μL of 1 mg/mL bovine serum albumin (Sigma, St. Louis, Missouri) . The precipitate is collected by vacuum filtration on glass fiber filters employing a TOMTEC™ filtration system and quantified by counting in a beta scintillation counter.

X-8951A FOR - 107 - Table 1 demonstrates the PKC selectivity of representative compounds in the above assays.

Table 1 IC50 (μΜ) Ex, PKC-a ?κς-β;_ ρκς-β2 PKA CaM CK-II src 1 1 0.05 0.04 NA NA >100 NA 2 4 0.4 0.2 >100 >100 >100 >100 3 0.3 0.03 0.02 >100 3 >100 >100 4 0.3 0.02 0.008 NA 3 >100 37 4s 1.3 0.048 0.033 >100 2.5 >100 63 4r 0.30 0.005 0.021 >100 0.69 . >100 33 5 0.28 0.012 0.005 >100 4.0 >100 21 5s 0.36 0.0047 0.0059 >100 8 >100 >100 5r 0.4 0.01 0.01 \ >100 5 >100 63 6 4.2 0.043 0.035 NA NA NA NA 7 >5.0 0.15 0.18 NA NA NA NA 8 2.5 0.037 0.032 NA NA NA NA 9 3.(h 0.35 0.16 >100 26 >100 58 11 5 0.3 0.1 >100 20 >100 >100 12 19 0.6 0.5 >100 93 >100 NA 13 >5.0 1.9 0.94 NA NA NA NA 15 >5.0 2.9 0.83 NA NA NA NA 16 >5.0 3.2 2.3 NA NA NA NA 17s 0.24 <0.005 <0.005 0.16 2.2 >100 NA 18s 6.4 0.38 0.30 >100 4.4 >100 >100 18r 3.4 0.083 0.087 >100 8.8 >100 NA 19r 0.48 0.032 0.030 >100 2.2 >100 >100 20r 0.89 0.04 0.03 >100 7.3 >100 74 20s 3 0.1 0.05 >100 .6.7 >100 16 21r 68 0.18 0.05 >100 56 >100 >100 21s >5.0 0.17 0.044 NA NA NA NA 23s 1.8 0.30 0.24 NA NA NA . NA 24s 3.5 0.49 0.38 NA NA NA NA 25r 94 0.043 0.12 >100 22 >100 NA 27 2.2 0.049 0.026 NA NA NA NA 28 1 0.07 0.08 NA 2 >100 >100 X-8951A FOR - 108 - 29 >100 0.7 0.8 NA NA NA NA 30 >100 1 2 >100 >10 >10 >100 31 0.3 0.02 0.03 >100 0.47 >100 >100 31r 0.24 0.019 0.008 NA NA NA NA 32 0.1 0.01 0.008 >100 0.9 >100 72 33 0.4. 0.05 0.04 NA 0.6 >100 61 34 1 0.1 0.1 NA 4 >100 >100 35 9 3 2 NA 82 >100 >100 36r 0.45 0.005 0.014 >100 7.1 >100 61 37 0.7 0.05 0.04 >100 5 >100 >100 38 4 0.2 0.1 >100 9 >100 >100 39 31 0.4 0.3 >100 >100 >100 >100 40 0.6 0.05 0.03 >100 5 >100 4.4 40s 0.4 0.03 0.02 >100 41 >100 NA 40r 0.30 0.01 0.01 · >100 8.0 >100 71 41 0.3 0.03 0.03 · NA 3 >100 91 42 >100 0.5 0.6 >100* >100 >100 >100 43 0.4 0.04 0.03 NA 0.6 >100 >100 44 >100 2 2 NA NA NA NA i 45 3 : 0.1 0.1 >100 39 >100 >125 46 3 0.04 0.04 >100 63 >100 >100 47 2 0.07 0.06 >100 70 >100 >125 48 >100 0.5 0.3 >100 >100 >100 >100 49 10 0.6 0.4 >100 >100 >100 >100 51 49 0.5 0.5 NA NA NA NA 54 0.16 0.005 0.004 NA NA NA NA 55 >5.0 0.41 0.38 NA NA NA NA NA - data are not available The compounds of the invention inhibit protein kinase C with an IC50 value of below 100 |im. In addition, the compounds of the invention selectively inhibit the beta-1 and beta-2 protein kinase C isozymes and have an IC50 value with respect to these isozymes of below 10 |lm.

As an inhibitor of protein kinase C, the compounds are useful in the treatment of conditions in which protein kinase C has demonstrated a role in the pathology. Conditions recognized X-8951A FOR - 10$ - 122092/2 in the art include: diabetes mellitus and its complicacions, ischemia, inflammation, central nervous system disorders, cardiovascular disease, Alzheimer's disease, dermatological disease and cancer.

Protein kinase C inhibitors have been shown to block inflammatory responses such as neutrophil oxidative burst, CD3 down-regulation in T-lymphocytes, and phorbol-induced paw edema. Twoemy, B. et al. Bin hgm. Biophvs . Res. Commiin. JJl: 1087-1092 (1990) ; Mulqueen, M.J. et al. Agents Actions 37: 85-89 (1992). Accordingly, as inhibitors of PKC, the present compounds are useful in treating inflammation.

Protein kinase C activity plays a central role in the functioning of the central nervous system. Huang, K.P. Trends Neurosci. 12: 425-432 (1989). In addition, protein kinase C inhibitors have been shown to prevent the damage seen in focal and central ischemic brain injury and brain edema. Hara, H. et al. .7. Cpreb. Blood Flow Metab. 10: 646-653 (1990); Shibata, S. et al. Brain Res. 594: 290-294 (1992). Recently, protein kinase C has been determined to be implicated in Alzheimer's disease. Shimohama, S. et al., Neurology 43: 1407-1413 (1993).

Accordingly, the compounds of the present invention are useful in treating Alzheimer's disease and ischemic brain injury.

Protein kinase C activity has long been associated with cell growth, tumor promotion and cancer. Rotenberg, S.A. and einstein, I.B. Biochem. Mol. Aspects Sel . Cancer 1: 25-73 (1991) . Ahmad et al.. Molecular Pharmacology; £2 858-862 (1993). It is known that inhibitors of protein kinase C are effective in preventing tumor growth in animals.

Meyer, T. et al. int. J. Cancer 43: 851-856 (1989); Akinagaka, S. et al. Cancer Res. 51: 4888-4892 (1991). The compounds of the present invention also act as multidrug reversal (MDR) agents making them effective compounds when administered in conjunction with other chemotherapeutic agents.

Protein kinase C activity also plays an important role in cardiovascular disease. Increased protein kinase C activity in the vasculature has been shown to cause increased vasoconstriction and hypertension. A known protein kinase C inhibitor prevented this increase. Bilder, G.E. et al. iLu X-8951A FOR - HO - Pharmacol. EXP. Ther. 252: 526-530 (1990) . Because protein kinase C inhibitors demonstrate inhibition of the neutrophil oxidative burst, protein kinase C inhibitors are also useful in treating cardiovascular ischemia and improving cardiac function following ischemia.. Muid, R.E. et al. FEBS Lett. 293: 169-172 (1990); Sonoki, H. et al. Kokvu-To unkan 37: 669-674 (1989) . The role of protein kinase C in platelet function has also been investigated and as shown elevated protein kinase C levels being correlated with increased response to agonists. Bastyr III, E.J. and Lu, J. Diabetes 4£: (Suppl. 1) 97A (1993) . PKC has been implicated in the biochemical pathway in the platelet-activity factor modulation of microvascular permeability.

Kobayashi et al., Amer. Phvs. Soc. H1214-H1220 (1994) . Potent protein kinase C inhibitors have been demonstrated to affect agonist-induced aggregation in platelets. Toullec, D. et al. J. Biol. Chem. 266: 15771-15781 (1991) . Protein kinase C inhibitors also block agonist-induced smooth- muscle cell proliferation. Matsumoto, H. and Sasaki, Y. Biochem. Biophvs.

Res. Commun. 158: 105-109 (1989) . Therefore, the present compounds are useful vin treating cardiovascular disease, atherosclerosis :and restenosis.

Abnormal activity of protein kinase C has also been linked to dermatological disorders such as psoriasis. Horn, F. et al. J. Invest. Dermatol. 88: 220-222 (1987); Raynaud, F. and Evain-Brion, D. Br. J. Dermatol. 124: 542-546 (1991) . Psoriasis is characterized by abnormal proliferation of keratinocytes .

Known protein kinase C inhibitors have been shown to inhibit keratinocyte proliferation in a manner that parallels their potency as PKC inhibitors. Hegemann, L. et al. Arch. Dermatol. Res. 283: 456-460 (1991); Bollag, W.B. et al. .T. Invest.

Dermatol. 100: 240-246 (1993) . Accordingly, the compounds as inhibitors of PKC are useful in treating psoriasis.

Protein kinase C has been linked to several different aspects of diabetes. Excessive activity of protein kinase C. has been linked to insulin signaling defects and therefore to the insulin resistance seen in Type II diabetes. Karasik, A. et al.

J. Biol. Chem. 265: 10226-10231 (1990); Chen, K.S. et al. Trans .

Assoc. Am. Physicians 104: 206-212 (1991) ; Chin, J.E. et al. J_<_ X-8951A FOR - 112 - Biol. Chem. 268: 6338-6347 (1993). In addition, studies have demonstrated a marked increase in protein kinase C activity in tissues known to be susceptible to diabetic complications when exposed to hyperglycemic conditions. Lee, T.-S. et al. J. Clin. Invest. 83: 90-94 (1989); Lee, T.-S. et al. Proc. Natl. Acad. Sci. USA 86: 5141-5145 (1989); Craven, P.A. and DeRubertis, F.R. J. Clin. nvest. 83: 1667-1675 (1989); Wolf, B.A. et al. i.

Clin. Invest. 87: 31-38 (1991); Tesfamariam, B. et al. J. Clin. Invest. 87: 1643-1648 (1991).

The compounds of the invention are also isozyme- selective. The compounds preferentially inhibit protein kinase C beta-1 and beta-2 isozyme over the protein kinase C isozymes, i.e., alpha, gamma, delta, epsilon, zeta, and eta. In general, the compounds demonstrate a minimum of a ten fold differential in the dosage required to inhibit P C beta-1 or beta-2 isozyme and the dosage required for equal inhibition of the alpha protein kinase C isozyme as measured in the PKC assay.

Accordingly, compounds of the present invention inhibit beta-1 and beta-2. isozymes of protein kinase C at much lower concentrations with minimal inhibition of the other PKC isozymes. This isozyme selectivity is demonstrated in Table 2 for a representative compound.

Table 2 Isozymes ED50 (μΜ) Compound (Ex) <* βΐ β2 γ δ ε 5 .28 0.019 0.005 .23 ' .31 1.0 Because of this selectivity, the compounds are particularly useful in treating those disease states in which protein kinase C isozyme beta-1 or beta-2 are associated. For example, the elevated blood glucose levels found in diabetes leads to an isozyme-specific elevation of the beta-2 isozyme in vascular tissues. Inoguchi et al., Proc. Natl. Acad. Sci. USA 89: 11059-11065 (1992) . A diabetes-linked elevation of the beta isozyme in human platelets has been correlated with their altered response to agonists. Bastyr III, E.J. and Lu, J.

Diabetes 42 : (Suppl 1) 97A (1993). The human vitamin D X-8951A FOR - 112, - receptor has been shown to be selectively phosphorylated by protein kinase C beta. This phosphorylation has been linked to alterations in the functioning of the receptor. Hsieh et al., Proc. Natl. Acad. Sci . USA 88: 9315-9319 (1991); Hsieh et al., J. Biol. Chem. 268: 15118-15126 (1993). In addition, recent work has shown that the beta-2 isozyme is responsible for erythroleukemia cell proliferation while the alpha isozyme is involved in megakaryocyte differentiation in these same cells. Murray et al., J. Biol. Chem. 268: 15847-15853 (1993).

The compounds of Formula I are preferably formulated prior to administration. Therefore, yet another embodiment of the present invention is a pharmaceutical formulation comprising a compound of Formula I and one or more pharmaceutically acceptable carriers, diluents or excipients.

The present pharmaceutical formulations are prepared by known procedures using well known ahd readily available ingredients. In making the compositions of the present invention, the active ingredient will usually be mixed with a carrier, or diluted by a carrier, or . enclosed within a carrier which may. be in the form of a capsule, sachet, paper or other container. When the carrier serves as a diluent, it may be a solid, semisolid or liquid material which acts as a vehicle, excipient or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosol (as a solid or in a liquid medium), soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

Some examples of suitable carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methyl cellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate and mineral oil. The formulations can additionally include lubricating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents. The compositions of the invention may be formulated so X-8951A FOR - 113 - as to provide quick, sustained or delayed release of the active ingredient after administration to the patient. The compositions are preferably formulated in a unit dosage form, each dosage containing from about 1 to about 500 mg, more usually about 5 to about 300 mg, of the active ingredient. However, it will be understood that the therapeutic dosage administered will be determined by the physician in the light of the relevant circumstances including the condition to be treated, the choice of compound to be administered and the chosen route of administration, and therefore the above dosage ranges are not intended to limit the scope of the invention in any way. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical carrier.

In addition to the above formulations, the compounds of the present invention may be administered topically. Topical formulations are ointments, creams, and gels.

Ointments generally are prepared using either (1) an oleaginous base, i.e., one consisting of fixed oils or hydrocarbons, such as white petrolatum or mineral oil, or (2) an absorbent base, i.e., one consisting of an anhydrous substance or substances which can absorb water, for example anhydrous lanolin. Customarily, following formation of the base, whether oleaginous or absorbent, the active ingredient (compound) is added to an amount affording the desired concentration.

Creams are oil/water emulsions. They consist of an oil phase (internal phase), comprising typically fixed oils, hydrocarbons, and the like, such as waxes, petrolatum, mineral oil, and the like, and an aqueous phase (continuous phase) , comprising water and any water-soluble substances, such as added salts. The two phases are stabilized by use of an emulsifying agent, for example, a surface active agent, such as sodium lauryl sulfate; hydrophilic colloids, such as acacia colloidal clays, veegum, and the like. Upon formation of the emulsion, the active ingredient (compound) customarily is added to an amount to achieve the desired concentration.

X-8951A FOR - m - Gels comprise a base selection from an oleaginous base, water, or an emulsion-suspension base. To the base is added a gelling agent which forms a matrix in the base, increasing its viscosity. Examples of gelling agents are hydroxypropyl cellulose, acrylic acid polymers, and the like. Customarily, the active ingredient (compounds) is added to the formulation at the desired concentration at a point preceding addition of the gelling agent.

The amount of compound incorporated into a topical formulation is not critical; the concentration should only be a range sufficient to permit ready application of the formulation to the an affected tissue area in an amount which will deliver the desired amount of compound.

The customary amount of a topical formulation to be applied to an affected tissue will depend upon an affected tissue size and concentration of compound in the formulation. Generally, the formulation will be applied to the effected tissue in an amount affording from about 1 to about 500 g compound per cm^ of an affected tissue. Preferably, the applied amount of compound will range from about 30 to about 300 μg cm2, more preferably, from about 50 to about 200 μg/cm2, and, most preferably, from about 60 to about 100 μg cm2.

The following formulation examples are illustrative only and are not intended to limit the scope of the invention in any way.

Formulation 1 Hard gelatin capsules are prepared using the following ingredients: * Quantity (mg/capsule ) Active agent 250 starch, dried 200 magnesium stearate 10 Total 460 mg The above ingredients are mixed and filled into hard gelatin capsules in 460 mg quantities.

X-8951A FOR - 115" - Formulation 2 A tablet is prepared using the ingredients below: Quant i ty (mg/capsule ) Active. agent cellulose, macrocrystalline silicon dioxide, fumed stearic acid Total The components are blended and compressed to form tablets weighing 665 mg.

Formulation 3 An aerosol solution is prepared containing the following components: v Quantity (mg/capsule ) Active agent ethanol Propellant 22 (chlorodifluoromethane) Total The active compound is mixed with ethanol. The mixture is added to a portion of the Propellant 22, cooled to -30°C and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted with the remainder of the propellant. The valve units are then fitted to the container.

X-8951A FOR - - Formulation 4 Tablets each containing 60 mg of active ingredient are made as follows : Quantity- dug/ capsule ) Active agent 60 mg starch 45 mg macrocrystalline cellulose 35 mg polyviny1pyrro1idone (as 10% solution in water) 4 mg sodium carboxymethyl starch 4.5 mg magnesium stearate 0.5 mg · talc 1 mg Total 150 mg The active ingredient, starch and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are theVi passed . through a No. 14 mesh U.S. sieve. The granules. so , produced are dried at 50°C and passed, through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 150 mg.

X-8951A FOR - 117 - Formulation 5 Capsules each containing 80 mg of medicament are made as follows: Quantity (mg/capsule ) Active agent 80 mg starch 59 mg microcrystalline cellulose 59 mg magnesium stearate 2 mg Total 200 mg The active ingredient, cellulose, starch and magnesium stearate are blended, passed through a No. 45 mesh U.S. sieve, and filled into hard gelatin capsules in 200 mg quantities.

Formulation 6 Suppositories each containing 225 mg of active ingredient may be made as follows : i Quantity (mg/capsule) Active agent 225 mg saturated fatty acid glycerides 2,000 mg Total 2,225 mg The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.

X-8951A FOR - 11$ - Formulation 7 Suspensions each containing 50 mg of medicament per 5 mL dose are made as follows : Quant i ty (mg/ ca sule ) Active agent 50 mg sodium carboxymethyl cellulose 50 mg syrup 1.25 mL benzoic acid solution 0.10 mL flavor q. v. color q.v. purified water to total 5 mL · The medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, flavor and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.

Formulation 8 An intravenous formulation may be prepared as follows: Quantity (mg/capsule) Active agent 250 mg isotonic saline 1000 mg The solution of the above ingredients is administered intravenously at a rate of 1 mL per minute to a subject in need of treatment.

Claims (1)

1. A process for compound of the V is or W is substituted or X and Y are independently substituted or together and combine to form is independently or R3 is R4R5 or R4 and R5 are independently is an amino n is independently or or a pharmaceutically acceptable salt or solvate comprising combining a mixture of a compound of the wherein and m are as defined a a concentration of about molar to about and an alkylating agent of the formula wherein X and Y are as defined above and L is a leaving at a concentration of about molar to about with about to about 10 equivalents of CS2CO3 at a rate of from about to about in a polar aprotic A process for preparing a compound of formula II as defined in claim comprising combining a compound of the formula V wherein X and Y are as defined above and is independently a leaving at a concentration of about 3 molar to with about to about 10 equivalents of Cs2C03 at a rate of from about to 5 about in a polar aprotic A compound of the formula 10 V is or W is substituted 15 or X and Y are independently substituted or together and W combine to form 20 is independently or R3 is or and R5 are independently or combine with the nitrogen to which they are bonded to form a saturated or unsaturated 5 or 6 member r 1 AA is an amino acid is independently or and n is independently or or a pharmaceutically acceptable salt or solvate A of Claim 3 substituted fused or is independently or R3 is or R4 and R5 are i dependently or combine with the nitrogen to which are bonded to a saturated or unsaturated or 6 ring or a pharmaceutically or solvate of which ia Z ia or hydrogen or R5 is combines the nitrogen to which they are bonded to farm a satarated or unsaturated 5 or 6 member p is independently 1 or is independently or or a pharmaceutically acceptable salt or solvate A compound of the m is 2 or is and V is or a phannaceutically acceptable salt or solvate A compound of the m is 2 or is and V is or a pharmaceuticaUy acceptable salt or soivate insufficientOCRQuality