Description of the Related Art Random screening of molecules for possible activity as therapeutic agents has occurred for many years and resulted in a number of important drug discoveries. While advances in molecular biology and computational chemistry have led to increased interest in what has been termed"rational drug design", such techniques have not proven as fast or reliable as initially predicted. Thus, in recent years there has been a renewed interest and return to random drug screening. To this end, particular strides having been made in new technologies based on the development of combinatorial chemistry libraries, and the screening of such libraries in search for biologically active members.

In general, combinatorial chemistry libraries are simply a collection of molecules. Such libraries vary by the chemical species within the library, as well as the methods employed to both generate the library members and identify which members interact with biological targets of interest. While this field is still young, methods for generating and screening libraries have already become quite diverse and sophisticated. For example, a recent review of various combinatorial chemical libraries has identified a number of such techniques (Dolle, J. Com. Chem., 2 (3): 383-433,2000), including the use of

both tagged and untagged library members (Janda, Proc. Natl. Acad. Sci. USA 91: 10779-10785, 1994).

Initially, combinatorial chemistry libraries were generally limited to members of peptide or nucleotide origin. To this end, the techniques of Houghten et al. illustrate an example of what is termed a"dual-defined iterative" method to assemble soluble combinatorial peptide libraries via split synthesis techniques (Nature (London) 354: 84-86,1991 ; Biotechniques 13: 412-421, 1992; Bioorg. Med. Chem. Lett. 3: 405-412,1993). By this technique, soluble peptide libraries containing tens of millions of members have been obtained.

Such libraries have been shown to be effective in the identification of opioid peptides, such as methionine-and leucine-enkephalin (Dooley and Houghten, Life Sci. 52,1509-1517, 1993), and a N-acylated peptide library has been used to identify acetalins, which are potent opioid antagonists (Dooley et al., Proc.

Natl. Acad. Sci. USA 90:10811-10815, 1993. More recently, an all D-amino acid opioid peptide library has been constructed and screened for analgesic activity against the mu (", u") opioid receptor (Dooley et al, Science 266: 2019-2022, 1994).

While combinatorial libraries containing members of peptide and nucleotide origin are of significant value, there is still a need in the art for libraries containing members of different origin. For example, traditional peptide libraries to a large extent merely vary the amino acid sequence to generate library members. While it is well recognized that the secondary structures of peptides are important to biological activity, such peptide libraries do not impart a constrained secondary structure to its library members.

To this end, some researchers have cyclized peptides with disulfide bridges in an attempt to provide a more constrained secondary structure (Tumelty et al., J. Chem. Soc. 1067-68, 1994; Eichler et al., Peptide Res. 7: 300-306,1994). However, such cyclized peptides are generally still quite flexible and are poorly bioavailable, and thus have met with only limited success.

More recently, non-peptide compounds have been developed which more closely mimic the secondary structure of reverse-turns found in biologically active proteins or peptides. For example, U. S. Pat. No. 5,440, 013 to Kahn and published PCT applications nos. W094/03494, W001/00210A1, and W001/16135A2 to Kahn each disclose conformationally constrained, non- peptidic compounds, which mimic the three-dimensional structure of reverse- turns. In addition, U. S. Pat. No. 5,929, 237 and its continuation-in-part U. S.

Pat. No. 6,013, 458, both to Kahn, disclose conformationally constrained compounds which mimic the secondary structure of reverse-turn regions of biologically active peptides and proteins. The synthesis and identification of conformationally constrained, reverse-turn mimetics and their application to diseases were well reviewed by Obrecht (Advances in Med. Chem. , 4, 1-68, 1999).

While significant advances have been made in the synthesis and identification of conformationally constrained, reverse-turn mimetics, there remains a need in the art for small molecules which mimic the secondary structure of peptides. There is also a need in the art for libraries containing such members, as well as techniques for synthesizing and screening the library members against targets of interest, particularly biological targets, to identify bioactive library members.

The present invention also fulfills these needs, and provides further related advantages by providing confomationally constrained compounds which mimic the secondary structure of reverse-turn regions of biologically active peptides and proteins.

Wnt signaling pathway regulates a variety of processes including cell growth, oncogenesis, and development (Moon et al., 1997, Trends Genet.

13,157-162 ; Miller et al., 1999, Oncogene 18,7860-7872 ; Nusse and Varmus, 1992, Cell 69,1073-1087 ; Cadigan and Nusse, 1997, Genes Dev. 11,3286- 3305; Peifer and Polais, 2000 Science 287,1606-1609 ; Polakis 2000, Genes Dev. 14,1837-1851). Wnt signaling pathway has been intensely studied in a

variety of organisms. The activation of TCF4/p-catenin mediated transcription by Wnt signal transduction has been found to play a key role in its biological functions (Molenaar et al., 1996, Cell 86: 391-399; Gat et al., 1998 Cell 95: 605- 614; Orford et al., 1999 J. Cell. Biol. 146: 855-868; Bienz and Clevers, 2000, Cell 103: 311-20).

In the absence of Wnt signals, tumor suppressor gene adenomatous polyposis coli (APC) simultaneously interacts with the serine kinase glycogen synthase kinase (GSK)-3p and p-catenin (Su et al., 1993, Science 262,1734-1737 : Yost et al., 1996 Genes Dev. 10,1443-1454 : Hayashi et al., 1997, Proc. Natl. Acad. Sci. USA, 94,242-247 : Sakanaka et al., 1998, Proc. Natl. Acad. Sci. USA, 95,3020-3023 : Sakanaka and William, 1999, J.

Biol. Chem 274,14090-14093). Phosphorylation of APC by GSK-3ß regulates the interaction of APC with p-catenin, which in turn may regulate the signaling function of ß-catenin (B. Rubinfeld et al., Science 272,1023, 1996). Wnt signaling stabilizes p-catenin allowing its translocation to the nucleus where it interacts with members of the lymphoid enhancer factor (LEF1)/T-cell factor (TCF4) family of transcription factors (Behrens et al., 1996 Nature 382,638-642 : Hsu et al., 1998, Mol. Cell. Biol. 18,4807-4818 : Roose et all., 1999 Science 285, 1923-1926).

Recently c-myc, a known oncogene, was shown to be a target gene for ß-catenin/TCF4-mediated transcription (He et al., 1998 Science 281 1509-1512: Kolligs et al., 1999 Mol. Cell. Biol. 19,5696-5706). Many other important genes, including cyclin D1, and metalloproteinase, which are also involved in oncogenesis, have been identified to be regulated by TCF4/bata- catenin transcriptional pathway (Crawford et al., 1999, Oncogene 18,2883- 2891: Shtutman et al., 1999, Proc. Natl. Acad. Sci. USA. , 11,5522-5527 : Tetsu and McCormick, 1999 Nature, 398,422-426).

Moreover, overexpression of several downstream mediators of Wnt signaling has been found to regulate apoptosis (Moris et al., 1996, Proc.

Natl. Acad. Sci. USA, 93,7950-7954 : He et al., 1999, Cell 99,335-345 : Orford

et al, 1999 J. Cell. Biol., 146,855-868 : Strove and Sussman, 1999, Exp. Cell.

Res. , 253,637-648). Overexpression of APC in human colorectal cancer cells induced apoptosis (Moris et al., 1996, Proc. Natl. Acad. Sci. USA. , 93,7950- 7954), ectopic expression of p-catenin inhibited apoptosis associated with loss of attachment to extracellular matrix (Orford et al, 1999, J. Cell Biol. 146,855- 868). Inhibition of TCF4/ß-catenin transcription by expression of dominant- negative mutant of TCF4 blocked Wnt-1-mediated cell survival and rendered cells sensitive to apoptotic stimuli such as anti-cancer agent (Shaoqiong Chen et al., 2001, J. Cell. Biol., 152,1, 87-96) and APC mutation inhibits apoptosis by allowing constitutive survivin expression, a well-known anti-apoptotic protein (Tao Zhang et al., 2001, Cancer Research, 62,8664-8667).

Although mutations in the Wnt gene have not been found in human cancer, a mutation in APC or p-catenin, as is the case in the majority of colorectal tumors, results in inappropriate activation of TCF4, overexpression of c-myc and production of neoplastic growth (Bubinfeld et al, 1997, Science, 275, 1790-1792: Morin et al, 1997, Science, 275,1787-1790 : Casa et al, 1999, Cell. Growth. Differ. 10,369-376). The tumor suppressor gene (APC) is lost or inactivated in 85% of colorectal cancers and in a variety of other cancers as well (Kinzler and Vogelstein, 1996, Cell 87,159-170). APC's principal role is that of a negative regulator of the Wnt signal transduction cascade. A center feature of this pathway involves the modulation of the stability and localization of a cytosolic pool of ß-catenin by interaction with a large Axin-based complex that includes APC. This interaction results in phosphorylation of p-catenin thereby targeting it for degradation.

CREB binding proteins (CBP) /p300 were identified initially in protein interaction assays, first through its association with the transcription factor CREB (Chrivia et al, 1993, Nature, 365,855-859) and later through its interaction with the adenoviral-transforming protein E1A (Stein et al., 1990, J.

Viol., 64,4421-4427 : Eckner et al., 1994, Genes. Dev. , 8,869-884). CBP had a potential to participate in variety of cellular functions including transcriptional

coactivator function (Shikama et al., 1997, Trends. Cell. Biol., 7,230-236 : Janknecht and Hunter, 1996, Nature, 383,22-23). CBP/p300 potentiates catenin-mediated activation of the siamois promoter, a known Wnt target (Hecht et al, 2000, EMBO J. 19,8, 1839-1850). ß-catenin interacts directly with the CREB-binding domain of CBP and p-catenin synergizes with CBP to stimulate the transcriptional activation of TCF4/ß-catenin (Ken-lchi Takemaru and Randall T. Moon, 2000 J. Cell. Biol., 149,2, 249-254).

BRIEF SUMMARY OF THE INVENTION From this background, it is seen that TCF4/p-catenin and CBP complex of Wnt pathway can be taken as target molecules for the regulation of cell growth, oncogenesis and apoptosis of cells, etc. Accordingly, the present invention addresses a need for compounds that block TCF4/ß-catenin transcriptional pathway by inhibiting CBP, and therefore can be used for treatment of cancer, especially colorectal cancer.

In brief, the present invention is directed to a new type of conformationally constrained compounds, which mimic the secondary structure of reverse-turn regions of biologically active peptides and proteins. This invention also discloses libraries containing such compounds, as well as the synthesis and screening thereof.

The compounds of the present invention have the following general formula (I) : wherein A is- (CHR3)- or- (C=O)-, B is- (CHR4)- or- (C=O)-, D is- (CHR5)- or- (C=O)-, E is- (ZR6)- or- (C=O)-, G is- (XR7) n-,- (CHR7)- (NR8)-,- (C=O)- (XR9)-, or- (C=O)-, W is-Y (C=O)-,- (C=O) NH-,- (S02)- or is absent, Y is oxygen, sulfur, or-NH-, X and Z is independently nitrogen or CH, n=0 or 1; and

Ri, R2, R3, R4, R5, R6, R7, R8 and R9 are the same or different and independently selected from an amino acid side chain moiety or derivative thereof, the remainder of the molecule, a linker and a solid support, and stereoisomers thereof.

In an embodiment wherein A is-(CHR3)-, B is- (C=O)-, D is- (CHR5)-, E is- (C=O)-, and G is- (XR7) n-, the compounds of this invention have the following formula (II) :

wherein W, X, Y and n are as defined above, and R1, R2, R3, R5 and R7 are as defined in the following detailed description.

In an embodiment wherein A is- (C=O)-, B is- (CHR4)-, D is -(C=O)-, E is- (ZR6)-, and G is- (C=O)- (XR9)-, the compounds of this invention have the following formula (III) :

wherein W, X and Y are as defined above, Z is nitrogen or CH (with the proviso that when Z is CH, then X is nitrogen), and Ri, R2, R4, R6 and Rg are as defined in the following detailed description.

In an embodiment wherein A is- (C=O)-, B is- (CHR4)-, D is- (C=O)-, E is- (ZR6)-, and G is (XR7) n-, the compounds of this invention have the following general formula (IV) :

wherein W, Y and n are as defined above, Z is nitrogen or CH (when Z is nitrogen, then n is zero, and when Z is CH, then X is nitrogen and n is not zero), and RI, R2, R4, R6 and R7, are as defined in the following detailed description.

The present invention is also directed to libraries containing one or more compounds of formula (I) above, as well as methods for synthesizing such libraries and methods for screening the same to identify biologically active compounds. Compositions containing a compound of this invention in combination with a pharmaceutical acceptable carrier or diluent are also disclosed.

The present invention is also related to methods for identifying a biologically active compound using the libraries containing one or more compound of formula (I). In a related aspect, the present invention provides a method for performing a binding assay, comprising (a) providing a composition comprising a first co-activator and an interacting protein, said first co-activator comprising a binding motif of LXXLL, LXXLI or FXXFF wherein X is any amino acid; (b) combining the first co-activator and the interacting protein with a test compound; and (c) detecting alteration in binding between the first co-activator and the interacting protein in the presence of the compound having general formula (I).

The present invention also provides methods for preventing or treating disorders associated with Wnt signaling pathway. Disorders that may be treated or prevented using a compound or composition of the present invention include tumor or cancer (e. g. , KSHV-associated tumor), restenosis associated with angioplasty, polycystic kidney disease, aberrant angiogenesis disease, rheumatoid arthritis disease, ulcerative colitis, tuberous sclerosis complex, hair loss, and Alzheimer's disease. Such methods comprise

administering to a subject in need thereof a compound or composition of the present invention in an amount effective to achieve the desired outcome.

In a related aspect, the present invention further provides methods for promoting neurite outgrowth, differentiation of a neural stem cell, and apoptosis in cancer cells. Such methods comprise administering to appropriate cells a compound or composition of the present invention in an amount effective to achieve the desired outcome.

These and other aspects of this invention will be apparent upon reference to the attached figure and following detailed description. To this end, various references are set forth herein, which describe in more detail certain procedures, compounds and/or compositions, and are incorporated by reference in their entirety.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 provides a general synthetic scheme for preparing reverse-turn mimetics of the present invention.

Figure 2 provides a general synthetic scheme for preparing reverse-turn mimetics of the present invention.

Figure 3 shows a graph based on the measurement of IC50 for Compound A of the present invention using SW480 cells, wherein cell growth inhibition on SW480 cells was measured at various concentrations of Compound A prepared in Example 4 to obtain the IC50 value. Specifically, the degree of inhibition in firefly and renilla luciferase activities by Compound A was determined. As a result, the IC5o of Compound A against SW480 cell growth was found as disclosed in Table 4. Detailed procedures are the same as disclosed in Example 6.

Figure 4. PC-12 cells were cultured on coated dishes, and differentiated for 10 days in 50 ng/ml nerve growth factor (NGF) (as described in Example 7). (A, B) Vector-transfected PC-12 cells (A) and PC-12 cells overexpressing wt PS-1 (B) exhibit extensive neurite outgrowth after 10 days in

NGF. (C) PC-12 cells expressing mutant PS-1/L286V do not display significant neurites under the same culture conditions. (D, E) Immunofluorescence analysis of GAP-43 (as described in Example 7), a molecular marker of neurite outgrowth, demonstrates intense staining for GAP-43 in the neurites (D) of vector-transfected and overexpressing PS-1/WT in PC-12 cells (E). (F) Lack of neurite outgrowth corresponds to weak GAP-43 immunostaining in the mutant cells. Data represent at least two independent experiments. (G) Differentiated cells were transfected with, Topflash, a TCF/R-catenin reporter construct. Cells were lysed, and luciferase activity measured 6 hours post-transfection (as described in Example 7). Data represent the mean of three independent experiments (SD). Asterisk indicate P < 0.05.

Fig. 5. Compound D phenotypically corrects deficient neuronal differentiation in PC-12 overexpressing mutant PS-1/L286V cells. Mutant cells were exposed to 10 uM Compound D, in addition to NGF, during the differentiation period (Misner et al., Proc. Natl. Acad. Sci. U S A 98,11714 (2001) ). (A) Neurite elongation and extension are observed in PC-12 cells overexpressing PS-1/L286V upon treatment with Compound D. (B) GAP-43 (green) is significantly elevated in the mutant cells, and is seen in the neurites.

(C) Quantitation of neurite outgrowth in PC-12 cells. Number of mutant cells with neurite lengths greater than two cell diameters was less than 10% that of the vector-transfected and overexpressing PS-1/WT in PC-12 cells. Number of mutant PS-1/L286V cells that had the defined neurite lengths was significantly increased, after treatment with 10 uM Compound D. The results are the average ( SD) of three independent determinations. Asterisk indicate P < 0.05.

Fig. 6. Ephrin B2 (EphB2) receptor expression.

Immunofluorescence analysis and RT-PCR were performed to detect EphB2 receptor expression (as described in Example 7). (A, B) EphB2 receptors are clearly demonstrated in neurites of vector-transfected and overexpressing PS- 1/WT cells. The intensity of staining correlates with the high expression level.

(C) In contrast, PS-1/L286V PC-12 cells have markedly reduced EphB2

receptor expression. (D) Treatment of mutant cells with Compound D leads to increased EphB2 receptor expression, which is focused at points of neurite outgrowth. (E) Expression of EphB2 receptor has previously been shown to be transcriptionally regulated (Guo et al., J. Neurosci. 17,4212 (1997). ). Lane 1, vector-transfected PC-12 cells, lane 2, overexpressing PS-1NVT cells, lane 3, overexpressing mutant PS-1/L286V cells, lane 4, mutant cells treated with Compound D. RT-PCR analysis indicates message for EphB2 receptor in cells overexpressing mutant PS-1/L286V is decreased compared to those in both the vector-transfected and overexpressing wt PS-1 PC-12 cells. Treatmentwith 10 uM Compound D upregulates EphB2 message. GAPDH is used an internal control.

Figure 7. A. Compound D arrests cells in G1. FACS analysis was performed on SW480 (lower panel) and HCT116 (upper panel) cells treated for 24 hours with either Compound D (25 pM) (right) or control (0. 5% DMSO (left).

5.5 X 106 cells were fixed and stained with propidium iodide (PI). B.

Compound D selectively activates caspases in colon carcinoma cell lines.

SW480 and HCT116 (left graph) cells (105) along with the normal colonocytes CCD18Co (right graph) were treated with either control (0.5% DMSO) or Compound D (25 ils). 24 hours post treatment, cells were lysed and the caspase-3/7 enzymatic activities were measured. Relative fluorescence units (RFU) were calculated by subtracting the unit values of the blank (control, without cells) from the treated samples (Compound D or control) and plotted.

Figure 8. Compound D reduces colony growth in soft agar in a dose dependent manner. Increasing concentrations of 5-fluorouracil (5-FU) (0. 5-321lu) and Compound D (0. 25-5 uM) were added to SW480 (5000 cells/well) of triplicate wells. Cells were washed and suspended in soft agar growth medium. The number of colonies after 8 days (colonies over 60 uM diameter) were counted and plotted against the compound concentration.

Mean SE of three determinations is indicated. The colony number of control in the absence of the compound was 1,637 + 71.

Figure 9. A. Compound C reduces tumor growth in nude mouse model. B. Compound C slightly reduces body weight in nude mouse model.

Figure 10. The survivin transcriptional activity is upregulated by Wnt1, but knout-down by Compound D. Percent luciferase activities were measured in wildtyp, CBP+/-, and p300+/-3T3 cells in the absence of Wnt1 and Compound D, or in the presence of Wnt1, Compound D or both.

Figure 11. Compound A (right graph) and Compound D (left graph) inhibit the activity of a survivin luciferase reporter in SW480 cells. The luciferase activities under the control of the survivin promoter were measured in SW480 cells treated with compound A or Compound D at various concentrations.

Figure 12. RT-PCR analsis indicates that Compound D treatment decreases the expression level of the survivin gene.

Figure 13. Compound D decreases the association of various proteins with the survivin promoter. ChIP assays on SW480 cells treated with either Compound D (25 uM) or control (0.5% DMSO) for 18 hours were performed.

Figure 14. Compound D decreases survivin expression at the translational level. A. Western blot analysis of extracts of cells treated with vehicle (0.5% DMSO) alone, 10 uM or 25 uM Compound D, or 5 uM 5-FU was performed using survivin 6E4 monoclonal antibody (Cell Signaling Technolgy).

B. Survivin immunofluorescence microscopy. Cultured cancer cells were fixed and stained with anti-survivin green. C. Survivin immunofluorescence microscopy. SW480 cells treated with Compound D were fixed and stained with anti-survivin green.

Figure 15. Compound D activates the caspase 3 activity (but not the caspase 2 activity) via suppression of the survivin expression. Cultured cells with or without transfection of a construct containing the survivin gene were treated with stausporine (0. 5 uM), Compound D (2. 5 uM or 5. 0 uM), or both. The caspase 2 and caspase 3 activities in these cells were measured.

Figure 16. Compound D promotes cell death via suppression of the survivin expression. Cultured cancer cells with or without transfection of a construct containing the survivin gene were treated with stausporine (0.5 pM), Compound D (5. 0 uM), or both. The cell death of these cells was measured.

Figure 17. Compound D increases the number of cells in Go.

Cultured cancer cells with or without transfection of a construct containing the survivin gene were treated with stausporine (0. 5 uM), Compound D (5 ils), or both. FACS analysis was performed on these cells and the percentages of cells in Go are indicated.

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to conformationally constrained compounds that mimic the secondary structure of reverse-turn regions of biological peptide and proteins (also referred to herein as"reverse-turn mimetics", and is also directed to chemical libraries relating thereto.

The reverse-turn mimetic structures of the present invention are useful as bioactive agents, including (but not limited to) use as diagnostic, prophylactic and/or therapeutic agents. The reverse-turn mimetic structure libraries of this invention are useful in the identification of bioactive agents having such uses. In the practice of the present invention, the libraries may contain from tens to hundreds to thousands (or greater) of individual reverse- turn structures (also referred to herein as"members").

In one aspect of the present invention, a reverse-turn mimetic structure is disclosed having the following formula (I) : wherein A is- (CHR3)- or- (C=O)-, B is- (CHR4)- or- (C=O)-, D is- (CHR5)- or- (C=O)-, E is- (ZR6)- or- (C=O)-, G is- (XR7) n-,- (CHR7)- (NR8)-,- (C=O)- (XR9)-,

or- (C=O)-, W is-Y (C=O)-,- (C=O) NH-,- (S02)- or nothing, Y is oxygen, sulfur, or-NH-, X and Z is independently nitrogen or CH, n=0 or 1; and Ri, R2, R3, R4, R5, R6, R7, R8 and Rg are the same or different and independently selected from an amino acid side chain moiety or derivative thereof, the remainder of the molecule, a linker and a solid support, and stereoisomers thereof.

In one embodiment, Ri, R2, R3, R4, R5, R6, R7, R8 and Rg are independently selected from the group consisting of aminoC25alkyl, guanidineC2-5alkyl, C1-4alkylguanidinoC2-5alkyl, diC1-4alkylguanidino-C2-5alkyl, amidinoC2-5alkyl, C1-4alkylamindoC2-5alkyl, diC1-4alkylamidinoC2-5alkyl, C1- 3alkoxy, phenyl, substituted phenyl (where the substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C1-4alkylamino, C1-4dialkylamino, halogen, perfluoro C1-4alkyl, C1- 4alkyl, C13alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), benzyl, substituted benzyl (where the substituents on the benzyl are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, Cl- 4alkylamino, C14dialkylamino, halogen, perfluoro C14alkyl, C13alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), naphthyl, substituted naphthyl (where the substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C14alkylamino, C14dialkylamino, halogen, perfluor C14alkyl, C14alkyl, C13alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), bis-phenyl methyl, substituted bis-phenyl methyl (where the substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C14alkylamino, C14dialkylamino, halogen, perfluoro C1-4alkyl, C1-4alkyl, C1-3alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), pyridyl, substituted pyridyl, (where the substituents are independently selected from one or more of amino amidino, guanidino, hydrazino, amidazonyl, C1-4alkylamino, C1-4dialkylamino, halogen, perfluoro C1-4alkyl, C1-4alkyl, C1- 3alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), pyridylC14alkyl, substituted pyridylCI-4alkyl (where the pyridine substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, Cl-

4alkylamino, C1-4dialkylamino, halogen, perfluoro C1-4alkyl, C1-4alkyl, C1-3alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), pyrimidylCl-4alkyl, substituted pyrimidylC-4alkyl (where the pyrimidine substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C1-4alkylamino, C1-4dialkylamino, halogen, perfluoro C1-4alkyl, C1-4alkyl, C1- 3alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), triazin-2-yi-CI-4alkyl, substituted triazin-2-yl-C14alkyl (where the triazine substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C14alkylamino, C14dialkylamino, halogen, perfluor C 4alkyl, C1-4alkyl, C1-3alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), imidazoC14alkyl, substituted imidazol C1-4alkyl (where the imidazole sustituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C1-4alkylamino, C1-4dialkylamino, halogen, perfluoro C1- 4alkyl, Cl-4alkyl, C13alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl), imidazolinC1-4alkyl, N-amidinopiperazinyl-N-C0-4alkyl, hydroxyC2-5alkyl, C1- 5alkylaminoC2-5alkyl, hydroxyC2-5alkyl, C1-5alkylaminoC2-5alkyl, Cl-5dialkylaminOC2-5alkyl, N-amidinopiperidinylCl-4alkyl and 4- aminocyclohexylC0-2alkyl.

In one embodiment, Ri, R2, R6 of E, and R7, R8 and R9 of G are the same or different and represent the remainder of the compound, and R3 of A, R4 of B or R5 of D is selected from an amino acid side chain moiety or derivative thereof. As used herein, the term"remainder of the compound" means any moiety, agent, compound, support, molecule, linker, amino acid, peptide or protein covalently attached to the reverse-turn mimetic structure at Ri, R2, R5, R6, R7, R8 and/or Rg positions. This term also includes amino acid side chain moieties and derivatives thereof.

In another embodiment R3 of A, Rs of D, R6 of E, and R7, R8, and Rg of G are the same or different and represent the remainder of the compound, while one or more of, and in one aspect all of, RI, R2 and R4 of B represent an amino acid sidechain. In this case, the term"remainder of the compound"

means any moiety, agent, compound, support, molecule, linker, amino acid, peptide or protein covalently attached to the reverse-turn mimetic structure at R3, R5, R6, R7, R8 and/or Rg positions. This term also includes amino acid side chain moieties and derivatives thereof.

As used herein, the term"remainder of the compound"means any moiety, agent, compound, support, molecule, atom, linker, amino acid, peptide or protein covalently attached to the reverse-turn mimetic structure. This term also includes amino acid side chain moieties and derivatives thereof. In one aspect of the invention, any one or more of the Ri, R2, R3, R4, R5, R6, R7, R8 and/or Rg positions may represent the remainder of the compound. In one aspect of the invention, one or more of Ri, R2 and R4 represents an amino acid side chain moiety or a derivative thereof.

As used herein, the term"amino acid side chain moiety" represents any amino acid side chain moiety present in naturally occurring proteins including (but not limited to) the naturally occurring amino acid side chain moieties identified in Table 1. Other naturally occurring amino acid side chain moieties of this invention include (but are not limited to) the side chain moieties of 3,5-dibromotyrosine, 3,5-diiodotyrosine, hydroxylysine, y- carboxyglutamate, phosphotyrosine and phosphoserine. In addition, glycosylated amino acid side chains may also be used in the practice of this invention, including (but not limited to) glycosylated threonine, serine and asparagine.

TABLE 1 Amino Acid Side Chain Moiety Amino Acid <BR> <BR> <BR> -H Glycine<BR> <BR> <BR> <BR> <BR> -CH3 Alanine -CH (CH3) 2 Valine - CH2 CH (CH3) 2 Leucine - CH (CH3) CH2 CH3 Isoleucine - (CH2) 4NH3+ Lysine - (CH2) 3NHC (NH2) NH2+ Arginine

CH2 1UNH Histidine -CH2COO-Aspartic acid -CH2CH2COO-Glutamic acid - CH2CONH2 Asparagine -CH2CH2CONH2 Glutamine CH2 Phenylalanine Tyrosine , CH21 < CHEZ H Tryptophan - CH2SH Cysteine -CH2CH2SCH3 Methionine - CH20H Serine - CH (OH) CH3 Threonine HNo/ Proline HNY OH f) H Hydroxyproline In addition to naturally occurring amino acid side chain moieties, the amino acid side chain moieties of the present invention also include various derivatives thereof. As used herein, a"derivative"of an amino acid side chain moiety includes modifications and/or variations to naturally occurring amino acid side chain moieties. For example, the amino acid side chain moieties of alanine, valine, leucine, isoleucine and phenylalanine may generally be classified as lower chain alkyl, aryl, or arylalkyl moieties. Derivatives of amino acid side chain moieties include other straight chain or branched, cyclic or

noncyclic, substituted or unsubstituted, saturated or unsaturated lower chain alkyl, aryl or arylalkyl moieties.

As used herein,"lower chain alkyl moieties"contain from 1-12 carbon atoms,"lower chain aryl moieties"contain from 6-12 carbon atoms and "lower chain aralkyl moieties"contain from 7-12 carbon atoms. Thus, in one embodiment, the amino acid side chain derivative is selected from a CI-12 alkyl, a C6-12 aryl and a 67-12 arylalkyl, and in a more preferred embodiment, from a Cri-7 alkyl, a C610 aryl and a 67-11 arylalkyl.

Amino side chain derivatives of this invention further include substituted derivatives of lower chain alkyl, aryl, and arylalkyl moieties, wherein the substituent is selected from (but is not limited to) one or more of the following chemical moieties:-OH,-OR,-COOH,-COOR,-CONH2,-NH2,-NHR, - NRR,-SH,-SR,-S02R,-S02H,-SOR and halogen (including F, Cl, Br and 1), wherein each occurrence of R is independently selected from straight chain or branched, cyclic or noncyclic, substituted or unsubstituted, saturated or unsaturated lower chain alkyl, aryl and aralkyl moieties. Moreover, cyclic lower chain alkyl, aryl and arylalkyl moieties of this invention include naphthalene, as well as heterocyclic compounds such as thiophene, pyrrole, furan, imidazole, oxazol, thiazole, pyrazol, 3-pyrroline, pyrrolidine, pyridine, pyrimidine, purine, quinoline, isoquinoline and carbazole. Amino acid side chain derivatives further include heteroalkyl derivatives of the alkyl portion of the lower chain alkyl and aralkyl moieties, including (but not limited to) alkyl and aralkyl phosphonates and silanes.

Representative Ri, R2, R3, R4, R5, R6, R7, R$ and R9 moieties specifically include (but are not limited to)-OH,-OR,-COR,-COOR,-CONH2,- CONR,-CONRR,-NH2,-NHR,-NRR,-S02R and-COSR, wherein each occurrence of R is as defined above.

In a further embodiment, and in addition to being an amino acid side chain moiety or derivative thereof (or the remainder of the compound in the case of Ri, R2, R3, R5, R6, R7, R8 and Rg), Ri, R2, R3, R4, R5, R6, R7, R$ or Rg

may be a linker facilitating the linkage of the compound to another moiety or compound. For example, the compounds of this invention may be linked to one or more known compounds, such as biotin, for use in diagnostic or screening assay. Furthermore, Ri, R2, R3, R4, Rs, R6, R7, R8 or Rg may be a linker joining the compound to a solid support (such as a support used in solid phase peptide synthesis) or alternatively, may be the support itself. In this embodiment, linkage to another moiety or compound, or to a solid support, is preferable at the Ri, R2, R7 or R8, or Rg position, and more preferably at the Ri or R2 position.

In the embodiment wherein A is- (CHR3)-, B is- (C=O)-, D is- (CHR5)-, E is- (C=O)-, and G is- (XR7) n-, the reverse turn mimetic compound of this invention has the following-formula (11) :

wherein R1, R2, R3, R5, R7, W, X and n are as defined above. In a preferred embodiment, R1, R2 and R7 represent the remainder of the compound, and R3 or R5 is selected from an amino acid side chain moiety.

In the embodiment wherein A is- (C=O)-, B is- (CHR4)-, D is- (C=O)-, E is- (ZR6)-, G is- (C=O)- (XR9)-, the reverse turn mimetic compound of this invention has the following general formula (III) :

wherein R1, R2, R4, R6, Rs, W and X are as defined above, Z is nitrogen or CH (when Z is CH, then X is nitrogen). In a preferred embodiment, Ri, R2, R6 and

Rg represent the remainder of the compound, and R4 is selected from an amino acid side chain moiety.

In a more specific embodiment wherein A is- (C=O)-, B is- (CHR4) -, D is- (C=O)-, E is- (ZR6)-, and G is (XR7) n-, the reverse turn mimetic compound of this invention has the following formula (IV) : wherein RI, R2, R4, R6, R7, W, X and n are as defined above, and Z is nitrogen or CH (when Z is nitrogen, then n is zero, and when Z is CH, then X is nitrogen and n is not zero). In a preferred embodiment, Ri, R2, R6 and R7 represent the remainder of the compound, and R4 is selected from an amino acid side chain moiety. In one aspect, R6 or R7 is selected from an amino acid side chain moiety when Z and X are both CH.

These compounds may be prepared by utilizing appropriate starting component molecules (hereinafter referred to as"component pieces").

Briefly, in the synthesis of reverse-turn mimetic structures having formula (I), first and second component pieces are coupled to form a combined first-second intermediate, if necessary, third and/or fourth component pieces are coupled to form a combined third-fourth intermediate (or, if commercially available, a single third intermediate may be used), the combined first-second intermediate and third-fourth intermediate (or third intermediate) are then coupled to provide a first-second-third-fourth intermediate (or first-second-third intermediate) which is cyclized to yield the reverse-turn mimetic structures of this invention.

Alternatively, the reverse-turn mimetic structures of formula (I) may be prepared by sequential coupling of the individual component pieces either stepwise in solution or by solid phase synthesis as commonly practiced in solid phase peptide synthesis.

Specific component pieces and the assembly thereof to prepare compounds of the present invention are illustrated in Figure 1. For example, a "first component piece"may have the following formula S1 : wherein R2 is as defined above, and R is a protective group suitable for use in peptide synthesis, where this protection group may be joined to a polymeric support to enable solid-phase synthesis. Suitable R groups include alkyl groups and, in a preferred embodiment, R is a methyl group. In Figure 1, one of the R groups is a polymeric (solid) support, indicated by"Pol"in the Figure.

Such first component pieces may be readily synthesized by reductive amination of H2N-R2 with CH (OR) 2-CHO, or by a displacement reaction between H2N-R2 and CH (OR) 2-CH2-LG (wherein LG refers to a leaving group, e. g. , a halogen (Hal) group).

A"second component piece"may have the following formula S2:

where P is an amino protection group suitable for use in peptide synthesis, Li is hydroxyl or a carboxyl-activation group, and R4 is as defined above. Preferred protection groups include t-butyl dimethylsilyl (TBDMS), t-butyloxycarbonyl (BOC), methyloxycarbonyl (MOC), 9H-fluorenylmethyloxycarbonyl (FMOC), and allyloxycarbonyl (Alloc). N-Protected amino acids are commercially available ; for example, FMOC amino acids are available from a variety of sources. In order for the second component piece to be reactive with the first component piece, Li is a carboxyl-activation group, and the conversion of carboxyl groups to activated carboxyl groups may be readily achieved by methods known in the art for the activation of carboxyl groups. Suitable activated carboxylic acid

groups include acid halides where Li is a halide such as chloride or bromide, acid anhydrides where Li is an acyl group such as acetyl, reactive esters such as an N-hydroxysuccinimide esters and pentafluorophenyl esters, and other activated intermediates such as the active intermediate formed in a coupling reaction using a carbodiimide such as dicyclohexylcarbodiimide (DCC).

Accordingly, commercially available N-protected amino acids may be converted to carboxylic activated forms by means known to one of skill in the art.

In the case of the azido derivative of an amino acid serving as the second component piece, such compounds may be prepared from the corresponding amino acid by the reaction disclosed by Zaloom et al. (J. Org.

Chem. 46: 5173-76,1981).

Alternatively, the first component piece of the invention may have the following formula S1': wherein R is as defined above and L2 is a leaving group such as halogen atom or tosyl group, and the second component piece of the invention may have the following formula S2' : wherein R2, R4 and P are as defined above, A"third component piece"of this invention may have the following formula S3:

where G, E, Li and L2 are as defined above. Suitable third component pieces are commercially available from a variety of sources or can be prepared by methods well known in organic chemistry.

In Figure 1, the compound of formula (1) has- (C=O)- for A,- (CHR4)-for B,- (C=O)- for D, and- (CR6)-for E. Compounds of formula (1) wherein a carbonyl group is at position B and an R group is at position B, i. e., compounds wherein A is- (CHR3)- and B is- (C=O)-, may be prepared in a manner analogous to that shown in Figure 1, as illustrated in Figure 2. Figure 2 also illustrates adding a fourth component piece to the first-second-third component intermediate, rather than attaching the fourth component piece to the third component piece prior to reaction with the first-second intermediate piece. In addition, Figure 2 illustrates the prepartion of compounds of the present invention wherein D is- (CHRe)- (rather than- (C=O)- as in Figure 1), and E is- (C=O)- (rather than- (CHR6)- as in Figure 1). Finally, Figure 2 illustrates the preparation of compounds wherein G is NR7.

Thus, as illustrated above, the reverse-turn mimetic compounds of formula (I) may be synthesized by reacting a first component piece with a second component piece to yield a combined first-second intermediate, followed by reacting the combined first-second intermediate with third component pieces sequentially to provide a combined first-second-third-fourth intermediate, and then cyclizing this intermediate to yield the reverse-turn mimetic structure.

The syntheses of representative component pieces of this invention are described in Preparation Examples and working Examples.

The reverse-turn mimetic structures of formula (III) and (IV) may be made by techniques analogous to the modular component synthesis disclosed above, but with appropriate modifications to the component pieces.

The reverse-turn mimetic structures of the present invention are useful as bioactive agents, such as diagnostic, prophylactic, and therapeutic agents. For example, the reverse-turn mimetic structures of the present invention may be used for modulating a cell signaling transcription factor related peptides in a warm-blooded animal, by a method comprising administering to the animal an effective amount of the compound of formula (I).

Further, the reverse-turn mimetic structures of the present invention may also be effective for inhibiting peptide binding to PTB domains in a warm-blooded animal ; for modulating G protein coupled receptor (GPCR) and ion channel in a warm-blooded animal ; for modulating cytokines in a warm- blooded animal.

Meanwhile, it has been found that the compounds of the formula (I), especially compounds of formula (VI) are effective for inhibiting or treating disorders modulated by Wnt-signaling pathway, such as cancer, especially colorectal cancer. wherein Ra is a phenyl group; a substituted phenyl group having one or more substituents wherein the one or more substituents are independently selected

from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, Cl- 4alkylamino, C1-4dialkylamino, halogen, perfluoro C1-4alkyl, C1-4alkyl, C1-3alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; a benzyl group; a substituted benzyl group with one or more substituents where the one or more substituents are independently selected from one or more of amino, amidino, guanidino, hydrazino, amidazonyl, C14alkylamino, C-4dialkylamino, halogen, perfluor CI-4alkyl, d-saikoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl group; or a bicyclic aryl group having 8 to 11 ring members, which may have 1 to 3 heteroatoms selected from nitrogen, oxygen or sulfur ; Rb is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen or sulfur, and aryl ring in the compound may have one or more substituents selected from a group consisting of halide, hydroxy, cyano, lower alkyl, and lower alkoxy groups; Rc is a saturated or unsaturated C16alkyl, C16alkoxy, perfluoro C16alkyl group; and Xi, X2, and X3 may be the same or different and independently selected from hydrogen, hydroxyl, and halide.

In another aspect, it is an object of the present invention to provide a pharmaceutical composition comprising a safe and effective amount of the compound having general formula (VI) and pharmaceutical acceptable carrier, which can be used for treatment of disorders modulated by Wnt signaling pathway, especially by TCF4-p-catenin-CBP complex.

Further, the present invention is to provide a method for inhibiting the growth of tumor cells by using the above-described composition of the present invention; a method for inducing apoptosis of tumor cells by using the above-described composition of the present invention; a method for treating a disorder modulated by TCF4-P catenin-CBP complex by using the above- described composition of the present invention; and a method of treating cancer such as colorectal cancer by administering the composition of the present invention together with other anti-cancer agent such as 5-fluorouracil (5-FU),

taxol, cisplatin, mitomycin C, tegafur, raltitrexed, capecitabine, and irinotecan, etc.

In a preferred embodiment of the present invention, the compound of the present invention has a (6S, 10R)-configuration as follows : wherein Ra and Rb have the same meanings as defined above.

In another aspect of this invention, libraries containing reverse- turn mimetic structures of the present invention are disclosed. Once assembled, the libraries of the present invention may be screened to identify individual members having bioactivity. Such screening of the libraries for bioactive members may involve; for example, evaluating the binding activity of the members of the library or evaluating the effect the library members have on a functional assay. Screening is normally accomplished by contacting the library members (or a subset of library members) with a target of interest, such as, for example, an antibody, enzyme, receptor or cell line. Library members which are capable of interacting with the target of interest, are referred to herein as"bioactive library members"or"bioactive mimetics". For example, a bioactive mimetic may be a library member which is capable of binding to an antibody or receptor, or which is capable of inhibiting an enzyme, or which is capable of eliciting or antagonizing a functional response associated, for example, with a cell line. In other words, the screening of the libraries of the present invention determines which library members are capable of interacting with one or more biological targets of interest. Furthermore, when interaction

does occur, the bioactive mimetic (or mimetics) may then be identified from the library members. The identification of a single (or limited number) of bioactive mimetic (s) from the library yields reverse-turn mimetic structures which are themselves biologically active, and thus are useful as diagnostic, prophylactic or therapeutic agents, and may further be used to significantly advance identification of lead compounds in these fields.

Synthesis of the peptide mimetics of the library of the present invention may be accomplished using known peptide synthesis techniques, in combination with the first, second and third component pieces of this invention.

More specifically, any amino acid sequence may be added to the N-terminal and/or C-terminal of the conformationally constrained reverse-turn mimetic. To this end, the mimetics may be synthesized on a solid support (such as PAM resin) by known techniques (see, e. g., John M. Stewart and Janis D. Young, Solid Phase Peptide Synthesis, 1984, Pierce Chemical Comp. , Rockford, ici.) or on a silyl-linked resin by alcohol attachment (see Randolph et al., J. Am Chem.

Soc. 117: 5712-14,1995).

In addition, a combination of both solution and solid phase synthesis techniques may be utilized to synthesize the peptide mimetics of this invention. For example, a solid support may be utilized to synthesize the linear peptide sequence up to the point that the conformationally constrained reverse- turn is added to the sequence. A suitable conformationally constrained reverse-turn mimetic structure which has been previously synthesized by solution synthesis techniques may then be added as the next"amino acid"to the solid phase synthesis (i. e., the conformationally constrained reverse-turn mimetic, which has both an N-terminus and a C-terminus, may be utilized as the next amino acid to be added to the linear peptide). Upon incorporation of the conformationally constrained reverse-turn mimetic structures into the sequence, additional amino acids may then be added to complete the peptide bound to the solid support. Alternatively, the linear N-terminus and C-terminus protected peptide sequences may be synthesized on a solid support, removed

from the support, and then coupled to the conformationally constrained reverse- turn mimetic structures in solution using known solution coupling techniques.

In another aspect of this invention, methods for constructing the libraries are disclosed. Traditional combinatorial chemistry techniques (see, e. g., Gallop et al., J. Med. Chem. 37: 1233-1251,1994) permit a vast number of compounds to be rapidly prepared by the sequential combination of reagents to a basic molecular scaffold. Combinatorial techniques have been used to construct peptide libraries derived from the naturally occurring amino acids.

For example, by taking 20 mixtures of 20 suitably protected and different amino acids and coupling each with one of the 20 amino acids, a library of 400 (i. e., 202) dipeptides is created. Repeating the procedure seven times results in the preparation of a peptide library comprised of about 26 billion (i. e., 208) octapeptides.

Specifically, synthesis of the peptide mimetics of the library of the present invention may be accomplished using known peptide synthesis techniques, for example, the General Scheme of [4,4, 0] Reverse-Turn Mimetic Library as follows : o o o o R2 R4 Step 1 Step 2 Br---NH /NHFmoc R4 YO - '0 Rz R4 0 Ry 0 Pol O Pol O RZ + H T Pol O NHFmoc R4 o O RZ Rq O R O p I I I R Ry Ste 3 P, O N\ NJ. NNJ, X Step 4a N N O R O T or ep j R"jt o H H or Step 4c N O LI. HO N OCH3 y H 0 (Y'=O, S or NH) Synthesis of the peptide mimetics of the libraries of the present invention was accomplished using a FlexChem Reactor Block which has 96 well plates by known techniques. In the above scheme'Pol'represents a

bromoacetal resin (Advanced ChemTech) and detailed procedure is illustrated below.

Step 1 A bromoacetal resin (37mg, 0. 98 mmol/g) and a solution of R2- amine in DMSO (1.4mL) were placed in a Robbins block (FlexChem) having 96 well plates. The reaction mixture was shaken at 60°C using a rotating oven [Robbins Scientific] for 12 hours. The resin was washed with DMF, MeOH, and then DCM Step 2 A solution of commercial available FmocAmino Acids (4 equiv.), PyBob (4 equiv.), HOAt (4 equiv. ), and DIEA (12 equiv. ) in DMF was added to the resin. After the reaction mixture was shaken for 12 hours at room temperature, the resin was washed with DMF, MeOH, and then DCM.

Step 3 To the resin swollen by DMF before reaction was added 25% piperidine in DMF and the reaction mixture was shaken for 30 min at room temperature. This deprotection step was repeated again and the resin was washed with DMF, Methanol, and then DCM. A solution of hydrazine acid (4 equiv. ), HOBt (4 equiv. ), and DIC (4 equiv. ) in DMF was added to the resin and the reaction mixture was shaken for 12 hours at room temperature. The resin was washed with DMF, MeOH, and then DCM.

Step 4a (Where hydrazine acid is MOC carbamate) The resin obtained in Step 3 was treated with formic acid (1.2 mL each well) for 18 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under a reduced pressure using SpeedVac

[SAVANT] to give the product as oil. The product was diluted with 50% water/acetonitrile and then lyophilized after freezing.

Step 4b (Where Fmoc hydrazine acid is used to make Urea through isocynate) To the resin swollen by DMF before reaction was added 25% piperidine in DMF and the reaction mixture was shaken for 30 min at room temperature. This deprotection step was repeated again and the resin was washed with DMF, Methanol, then DCM. To the resin swollen by DCM before reaction was added isocynate (5 equiv. ) in DCM. After the reaction mixture was shaken for 12 hours at room temperature the resin was washed with DMF, MeOH, then DCM. The resin was treated with formic acid (1.2 mL each well) for 18 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under a reduced pressure using SpeedVac [SAVANT] to give the product as oil. The product was diluted with 50% water/acetonitrile and then lyophilized after freezing.

Step 4c (Where Fmoc-hydrazine acid is used to make Urea through active carbamate) To the resin swollen by DMF before reaction was added 25% piperidine in DMF and the reaction mixture was shaken for 30 min at room temperature. This deprotection step was repeated again and the resin was washed with DMF, MeOH, and then DCM. To the resin swollen by DCM before reaction was added p-nitrophenyl chloroformate (5 equiv. ) and diisopropyl ethylamine (5 equiv. ) in DCM. After the reaction mixture was shaken for 12 hours at room temperature, the resin was washed with DMF, MeOH, and then DCM. To the resin was added primary amines in DCM for 12 hours at room temperature and the resin was washed with DMF, MeOH, and then DCM. After reaction the resin was treated with formic acid (1.2 mL each well) for 18 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under a reduced pressure using SpeedVac

[SAVANT] to give the product as oil. The product was diluted with 50% water/acetonitrile and then lyophilized after freezing.

To generate these block libraries the key intermediate hydrazine acids were synthesized according to the procedure illustrated in Preparation Examples.

Tables 2A and 2B show a [4, 4, 0] Reverse turn mimetics library which can be prepared according to the present invention, of which representative preparation is given in Example 4.

TABLE 2A THE H 4, 0]REVERSE TURN MIMETICS LIBRARY No R2 R4 Rz Ri-Y'Mol. Weight M+H 1 2, 4-CI2-benzyl 4-HO-benzyl Allyl OCH3 533 534 2 2, 4-CI2-benzyl 4-NO2-benzyl Allyl OCH3 562 563 3 2, 4-CI2-benzyl 2, 4-F2-benzyl Allyl OCH3 553 554 4 2, 4-CI2-benzyl 4-CI-benzyl Allyl OCH3 552 553 5 2, 4-Cl2-benzyl 2, 2-bisphenylethyl Allyl OCH3 594 595 6 2, 4-CI2-benzyl 3-t-Bu-4-HO-benzyi Allyl OCH3 590 591 7 2, 4-CI2-benzyl 4-Me-benzyl Allyl OCH3 531 532 8 2,4-Cl2-benzyl Cyclohexylmethyl Allyl OCH3 523 524 9 2, 4-CI2-benzyl 4-F-benzyl Allyi OCHs 535 536 10 2, 4-CI2-benzyl 2-CI-benzyl Allyl OCH3 552 553 11 2, 4-Cl2-benzyl 2, 4-Cl2-benzyl Allyl OCH3 586 587 12 2, 4-CI2-benzyl Naphth-2-ylmethyl Allyl OCH3 567 568 13 2, 4-Cl2-benzyl 4-HO-benzyl Benzyl OCH3 583 584 14 2, 4-Cl2-benzyl 4-NO2-benzyl Benzyl OCH3 612 613 15 2, 4-Cl2-benzyl 2, 4-F2-benzyl Benzyl OCH3 603 604 16 2, 4-Cl2-benzyl 4-CI-benzyl Benzyl OCHs 602 603 17 2, 4-CI2-benzyl 2, 2-bisphenylethyl Benzyl OCH3 644 645 18 2, 4-CI2-benzyl 3-t-Bu-4-HO-benzy Benzyl OCH3 640 641 19 2, 4-CI2-benzyl 4-Me-benzyl Benzyl OCH3 582 583 20 2, 4-Cl2-benzyl Cyclohexylmethyl Benzyl OCH3 574 575 21 2,4-Cl2-benzyl 4-F-benzyl Benzyl OCH3 585 586 22 2,4-Cl2-benzyl 2-Cl-benzyl Benzyl OCH3 602 603 23 2, 4-Cl2-benzyl 2, 4-Cl2-benzyl Benzyl OCH3 636 637 No R2 R4 R7 R1-Y' Mol. Weight M+H 24 2, 4-CI2-benzyl Naphth-2-ylmethyl Benzyl OCH3 618 619 25 2, 4-Cl2-benzyl 4-HO-benzyl Allyl OCH3 479 480 26 2, 4-CI2-benzyl 4-NO2-benzyl Allyl OCH3 508 509 27 2, 4-CI2-benzyl 2, 4-F2-benzyl Allyl OCH3 499 500 28 2, 4-CI2-benzyl 4-CI-benzyl Allyl OCH3 497 498 29 Phenethyl 2, 2-bisphenylethyl Allyl OCH3 539 540 30 Phenethyl 3-t-Bu-4-HO-benzy Allyl OCH3 535 536 31 Phenethyl 4-Me-benzyl Allyl OCH3 477 478 32 Phenethyl Cyclohexylmethyl Allyl OCH3 469 470 33 Phenethyl 4-F-benzyl Allyl OCH3 481 482 34 Phenethyl 2-Cl-benzyl Allyl OCH3 497 498 35 Phenethyl 2, 4-CI2-benzyl Allyl OCH3 531 532 36 Phenethyl Naphth-2-ylmethyl Allyl OCHs 513 514 37 Phenethyl 4-HO-benzyl Benzyl OCH3 529 530 38 Phenethyl 4-NO2-benzyl Benzyl OCH3 558 559 3 Phenethyl 2, 4-F2-benzyl Benzyl OCH3 549 550 40 Phenethyl 4-Cl-benzyl Benzyl OCH3 547 548 41 Phenethyl 2, 2-bisphenylethyl Benzyl OCH3 589 590 42 Phenethyl 3-t-Bu-4-HO-benzy Benzyl OCH3 585 586 43 Phenethyl 4-Me-benzyl Benzyl OCH3 527 528 44 Phenethyl Cyclohexyl-methyl Benzyl OCH3 519 520 45 Phenethyl 4-F-benzyl Benzyl OCH3 531 532 46 Phenethyl 2-Cl-benzyl Benzyl OCH3 547 548 47 Phenethyl 2, 4-CI2-benzyl Benzyl OCH3 582 583 48 Phenethyl Naphth-2-ylmethyl Benzyl OCH3 563 564 49 Phenethyl 4-HO-benzyl Allyl OCH3 497 498 50 Phenethyl 4-NO2-benzyl Allyl OCH3 526 527 51 Phenethyl 2, 4-F2-benzyl Allyl OCH3 517 518 52 Phenethyl 4-Cl-benzyl Allyl OCH3 515 516 53 4-F-phenylethyl 2, 2-bisphenylethyl Allyl OCH3 557 558 54 4-F-phenylethyl 3-t-Bu-4-HO-benzy Allyl OCH3 553 554 55 4-F-phenylethyl 4-Me-benzyl Allyl OCHs 495 496 56 4-F-phenylethyl Cyclohexyl-methyl Allyl OCH3 487 488 57 4-F-phenylethyl 4-F-benzyl Allyl OCH3 499 500 58 4-F-phenylethyl 2-Cl-benzyl Allyl OCH3 515 516 59 4-F-phenylethyl 2, 4-Cl2-benzyl Allyl OCH3 549 550 60 4-F-phenylethyl Naphth-2-ylmethyl Allyl OCH3 531 532 61 4-F-phenylethyl 4-HO-benzyl Benzyl OCH3 547 548 62 4-F-phenylethyl 4-NO2-benzyl Benzyl OCH3 576 577 63 4-F-phenylethyl 2, 4-F2-benzyl Benzyl OCH3 567 568 64 4-F-phenylethyl 4-CI-benzyl Benzyl OCH3 565 566 65 4-F-phenylethyl 2, 2-bisphenylethyl Benzyl OCH3 607 608 66 4-F-phenylethyl 3-t-Bu-4-HO-benzyl Benzyl OCH3 603 604 67 4-F-phenylethyl 4-Me-benzyl Benzyl OCH3 545 546 68 4-F-phenylethyl Cyclohexyl-methyl Benzyl OCH3 537 538 69 4-F-phenylethyl 4-F-benzyl Benzyl OCH3 549 550 70 4-F-phenylethyl 2-CI-benzyl Benzyl OCH3 565 566 71 4-F-phenylethyl 2, 4-Cl2-benzyl Benzyl OCH3 599 600 72 4-F-phenylethyl Naphth-2-ylmethyl Benzyl OCH3 581 582 No R2 R4 R7 Ri-Y'Mol. Weight M+H 73 4-F-phenylethyl 4-HO-benzyl Allyl OCH3 509 510 74 4-F-phenylethyl 4-NO2-benzyl Allyl OCH3 538 539 75 4-F-phenylethyl 2, 4-F2-benzyl Allyl OCH3 529 530 76 4-F-phenylethyl 4-CI-benzyl Allyl OCH3 527 528 77 4-MeO-2, 2-bisphenylethyl Allyl OCH3 569 570 phenylethyl 78 4-MeO-3-t-Bu-4-HO-benzy Ailyl OCH3 565 566 phenylethyl 79 4-MeO-4-Me-benzyl Allyl OCH3 507 508 phenylethyl 80 4-MeO-Cyclohexyl-methyl Allyl OCH3 499 500 phenylethyl 81 4-MeO-4-F-benzyl Allyl OCH3 511 512 phenylethyl 82 4-MeO- 2-Cl-benzyl Allyl OCH3 527 528 phenylethyl 83 4-MeO- 2,4-Cl2-benzyl Allyl OCH3 561 562 phenylethyl 84 4-MeO-Naphth-2-ylmethyl Allyl OCH3 543 544 phenylethyl 85 4-MeO- 4-HO-benzyl Benzyl OCH3 559 560 phenylethyl 86 4-MeO-4-NO2-benzyl Benzyl OCH3 588 589 phenylethyl 87 4-MeO 2,4-F2-benzyl Benzyl OCH3 579 580 phenylethyl 88 4-MeO 4-Cl-benzyl Benzyl OCH3 577 578 phenylethyl 89 4-MeO-2, 2-bisphenylethyl Benzyl OCH3 619 620 phenylethyl 90 4-MeO- 3-t-Bu-4-HO-benzyl Benzyl OCH3 615 616 phenylethyl 91 4-MeO- 4-Me-benzyl Benzyl OCH3 557 558 phenylethyl 92 4-MeO-Cyclohexylmethyl Benzyl OCH3 549 550 phenylethyl 93 4-MeO-4-F-benzyl Benzyl OCH3 561 562 phenylethyl 94 4-MeO-2-CI-benzyl Benzyl OCH3 577 578 phenylethyl 95 4-MeO-2, 4-CI2-benzyl Benzyl OCH3 612 613 phenylethyl 96 4-MeO-Naphth-2-ylmethyl Benzyl OCH3 593 594 phenylethyl 97 Isoamyl 4-HO-benzyl Styrylmethyl OCH3 521 522 98 Isoamyl 4-NO2-benzyl Styrylmethyl OCH3 550 551 99 Isoamyl 2, 4-F2-benzyl Styrylmethyl OCH3 541 542 10 Isoamyl 4-CI-benzyl Styrylmethyl OCH3 539 540 101 Isoamyl 2, 2-bisphenylethyl Styrylmethyl 581 582 102 Isoamyl 3-t-Bu-4-HO-benzy Styrylmethyl OCH3 497 498 10 Isoamyl 4-Me-benzyl Styrylmethyl OCH3 519 520 104 Isoamyl Cyclohexylmethyl Styrylmethyl OCH3 511 512 10 Isoamyl 4-F-benzyl Styrylmethyl OCH3 523 524 No R2 R4 R7 R1-Y' Mol. Weight M+H 106 Isoamy I 2-Clbenzyl Styrylmethyl OCH3 539 540 10 Isoamyl 2, 4-Cl2-benzyl Styrylmethyl OCH3 574 575 10 Isoamyl Naphth-2-ylmethyl Styrylmethyl OCH3 555 556 10 Isoamyl 4-HO-benzyl 2, 6-Cl2- OCH3 563 564 benzyl 110 Isoamyl 4-NO2-benzyl 2, 6-Cl2- OCH3 592 593 benzyl 111 Isoamyl 2, 4-F2-benzyl 2, 6-Cl2- OCH3 583 584 benzyl 11 Isoamyl 4-CI-benzyl 2, 6-Cb- OCHs 582583 benzyl 11 Isoamyl 2,2-bisphenylethyl 2,6-Cl2- OCH3 624 625 benzyl 114 Isoamyl 3-t-Bu-4-HO-benzyl 2,6-Cl2- OCH3 540 541 benzyl 115 Isoamyl 4-Me-benzyl 2, 6-Cb-OCHs562563 benzyl 11 Isoamyl Cyclohexylmethyl 2, 6-Cl2- OCH3 554 555 benzyl 117 Isoamyl 4-F-benzyl 2,6-Cl2- OCH3 565 566 benzyl 11 Isoamyl 2-Cl-benzyl 2,6-Cl2- OCH3 582 583 benzyl 119 Isoamyl 2,4-Cl2-benzyl 2, 6-Cl2- OCH3 616 617 benzyl, 120 Isoamyl Naphth-2-ylmethyl 2,6-Cl2- OCH3 598 599 benzyl 121 3-MeO-propyl 4-HO-benzyl Styrylmethyl OCH3 523 524 12 3-MeO-propyl 4-NO2-benzyl Styrylmethyl OCH3 552 553 12 3-MeO-propyl 2, 4-F2-benzyl Styrylmethyl OCH3 543 544 12 3-MeO-propyl 4-CI-benzyl Styrylmethyl OCH3 541 542 12 3-MeO-propyl 2, 2-bisphenylethyl Styrylmethyl OCH3 583 584 12 3-MeO-propyl 3-t-Bu-4-HO-benzy Styrylmethyl OCH3 499 500 12 3-MeO-propyl 4-Me-benzyl Styrylmethyl OCH3 521 522 12E 3-MeO-propyl Cyclohexyl-methyl Styrylmethyl OCH3 513 514 129 3-MeO-propyl 4-F-benzyl Styrylmethyl OCH3 525 526 13 3-MeO-propyl 2-Cl-benzyl Styrylmethyl OCH3 541 542 131 3-MeO-propyl 2, 4-Cl2-benzyl Styrylmethyl OCH3 575 576 13 3-MeO-propyl Naphth-2-ylmethyl Styrylmethyl OCH3 557 558 13 3-MeO-propyl 4-HO-benzyl 2, 6-Cl2- OCH3 565 566 benzyl 134 3-MeO-propyl 4-NO2-benzyl 2,6-Cl2 OCH3 594 595 benzyl 13 3-MeO-propyl 2, 4-F2-benzyl 2, 6-Cl2 OCH3 565 586 benzyl 136 3-MeO-propyl 4-Cl-benzyl 2,6-Cl2- OCH3 584 585 benzyl 13 3-MeO-propyl 2, 2-bisphenylethyl 2, 6-CI2- OCH3 626 627 benzyl 13 3-MeO-propyl 3-t-Bu-4-HO-benzy 2, 6-CI2- OCH3 541 542 benzyl 13 3-MeO-propyl 4-Me-benzyl 2, 6-Cl2- OCH3 563 564 benzyl No R2 R4 R7 R1-Y' Mol. Weight H+A 140 3-MeO-propyl Cyclohexyl-methyl 2, 6-Cl2- OCH3 556 557 benzyl 141 3-MeO-propyl 4-F-benzyl 2,6-Cl2- OCH3 567 568 benzyl 14 3-MeO-propyl 2-Cl-benzyl 2,6-Cl2- OCH3 584 585 benzyl 143 3-MeO-propyl 2, 4-CI2-benzyl 2, 6-Cl2- OCH3 618 619 benzyl 14 3-MeO-propyl Naphth-2-ylmethyl 2, 6-Cl2- OCH3 600 601 benzyl 14 4-MeO-4-HO-benzyl Styrylmethyl OCH3 585 586 phenylethyl 146 4-MeO- 4-NO2-benzyl Styrylmethyl OCH3 614 615 phenylethyl 14 4-MeO-2, 4-F2-benzyl Styrylmethyl OCH3 605 606 phenylethyl 14 4-MeO-4-CI-benzyl Styrylmethyl OCH3 603 604 phenylethyl 14 4-MeO-2, 2-bisphenylethyl Styrylmethyl OCH3 645 646 phenylethyl 15 4-MeO-3-t-Bu-4-HO-benzy Styrylmethyl OCH3 561 562 phenylethyl 151 4-MeO-4-Me-benzyl Styrylmethyl OCH3 583 584 phenylethyl 15 4-MeO- Cyclohexyl-methyl Styrlmethyl OCH3 575 576 phenylethyl 15 4-MeO- 4-F-benzyl Styrylmethyl OCH3 587 588 phenylethyl 15 4-MeO-2-CI-benzyl Styrylmethyl OCH3 603 604 phenylethyl 15 4-MeO-2, 4-CI2-benzyl Styrylmethyl OCH3 638 639 phenylethyl 15 4-MeO- Naphth-2-ylmethyl Styrylmethyl OCH3 619 620 phenylethyl 15 4-MeO- 4-HO-benzyl 2, 6-Cl2- OCH3 628 629 phenylethyl benzyl 156 4-MeO- 4-NO2-benzyl 2, 6-Cl2- OCH3 657 658 phenylethyl benzyl 15 4-MeO-2, 4-F2-benzyl 2,6-Cl2- OCH3 648 649 phenylethyl benzyl 16 4-MeO-4-CI-benzyl 2, 6-CI2- OCHs 646 647 phenylethyl benzyl 161 4-MeO-2, 2-bisphenylethyl 2, 6-Cl2- OCH3 688 689 phenylethyl benzyl 162 4-MeO- 3-t-Bu-4-HO-benzyl 2,6-Cl2- OCH3 604 605 phenylethyl benzyl 163 4-MeO- 4-Me-benzyl 2,6-Cl2- OCH3 626 627 phenylethyl benzyl 16 4-MeO- Cyclohexylmethyl 2,6-Cl2- OCH3 618 619 phenylethyl benzyl 165 4-MeO- 4-F-benzyl 2,6-Cl2- OCH3 630 631 phenylethyl benzyl 16 4-MeO-2-CI-benzyl 2, 6-Cl2- OCH3 646 647 phenylethyl benzyl No R2 R4 R7 R1-Y' Mol. Weight M+H 16 4-MeO- 2, 4-Cl2-benzyl 2, 6-Cl2- OCH3 680 681 phenylethyl benzyl 168 4-MeO- Naphth-2-ylmethyl 2,6-Cl2 OCH3 662 663 phenylethyl benzyl 16 Tetrahydrofuran-4-HO-benzyl Styrylmethyl OCH3 535 536 2-ylmethyl 17 Tetrahydrofuran-4-NO2-benzyl Styrylmethyl OCH3 564 565 2-ylmethyl 171 Tetrahydrofuran-2, 4-F2-benzyl Styrylmethyl OCH3 555 556 2-ylmethyl 17 Tetrahydrofuran- 4-Cl-benzyl Styrylmethyl OCH3 553 554 2-ylmethyl 17 Tetrahydrofuran-2, 2-bisphenylethyl Styrylmethyl OCH3 595 596 2-ylmethyl 17 Tetrahydrofuran-3-t-Bu-4-HO-benzy Styrylmethyl OCH3 511 512 2-ylmethyl 17 Tetrahydrofuran-4-Me-benzyl Styrylmethyl OCH3 533 534 2-ylmethyl 17 Tetrahydrofuran-Cyclohexyl-methyl Styrylmethyl OCH3 525 526 2-ylmethyl 17 Tetrahydrofuran- 4-F-benzyl Styryfmethyl OCH3 537 538 2-ylmethyl 17 Tetrahydrofuran-2-CI-benzyl Styrylmethyl OCH3 553 554 2-ylmethyl 17 Tetrahydrofuran-2, 4-CI2-benzyl Styrylmethyl OCH3 588 589 2-ylmethyl 18 Tetrahydrofuran- Naphth-2-ylmethyl Styrylmethyl OCH3 569 570 2-ylmethyl 181 Tetrahydrofuran- 4-HO-benzyl 2,6-Cl2- OCH3 577 578 2-ylmethyl benzyl 18 Tetrahydrofuran- 4-NO2-benzyl 2,6-Cl2- OCH3 606 607 2-ylmethyl benzyl 18 Tetrahydrofuran-2, 4-F2-benzyl 2, 6-Cl2- OCH3 597 598 2-ylmethyl benzyl 18 Tetrahydrofuran- 4-Cl-benzyl 2, 6-CI2- OCHs 596 597 2-ylmethyl benzyl 18 Tetrahydrofuran-2, 2-bisphenylethyl 2, 6-Cl2- OCH3 638 639 2-ylmethyl benzyl 186 Tetrahydrofuran- 3-t-Bu-4-HO-benzyl 2,6-Cl2- OCH3 553 554 2-ylmethyl benzyl 18 Tetrahydrofuran- 4-Me-benzyl 2, 6-Cl2- OCH3 575 576 2-ylmethyl benzyl 18 Tetrahydrofuran- Cyclohexyl-methyl 2,6-Cl2- OCH3 568 569 2-yimethyl benzyl 18 Tetrahydrofuran-4-F-benzyl 2, 6-Cl2- OCH3 579 580 2-ylmethyl benzyl 19 Tetrahydrofuran- 2-Cl-benzyl 2,6-Cl2- OCH3 596 597 2-ylmethyl benzyl 191 Tetrahydrofuran- 2,4-Cl2-benzyl 2, 6-Cl2- OCH3 630 631 2-ylmethyl benzyl 19 Tetrahydrofuran-Naphth-2-ylmethyl 2, 6-CI2-OCH3 612 613 2-ylmethyl benzyl 19 Phenethyl 4-HO-benzyl Methyl (4-Me-528 529 phenyl) amino No R2 R4 R7 Ri-Y'Mol. Weight M+H 194 Phenethyl 4-HO-benzyl Methyl (4-CI-548 549 phenyl) amino 19 Phenethyl 4-HO-benzyl Methyl Phenylamino 514 515 19 Phenethyl 4-HO-benzyl Methyl ((R)-a-542 543 methylbenzyl) amino 197 Phenethyl 4-HO-benzyl Methyl Benzylamino 528 529 198 Phenethyl 4-HO-benzyl Methyl (4-MeO- 544 545 phenyl) amino 19 Phenethyl 4-HO-benzyl Methyl (4-Br-592 593 phenyl) amino 20 Phenethyl 4-HO-benzyl Methyl (4-CFs-582 583 phenyl)amino 201 Phenethyl 4-HO-benzyl Methyl Pentylamino 508 509 202 Phenethyl 4-HO-benzyl Methyl (2- 542 543 Phenylethyl) amino 20 Phenethyl 4-HO-benzyl Methyl (4-MeO-558 559 benzyl) amino 204 Phenethyl 4-HO-benzyl Methyl I Cyclohexylami 520 521 no 205 2,2- 4-HO-benzyl Methyl (4-Me- 604 605 bisphenylethyl phenyl) amino 20 2, 2- 4-HO-benzyl Methyl (4-Cl- 624 625 bisphenylethyl phenyl) amino 207 2, 2- 4-HO-benzyl Methyl Phenylamino 590 591 bisphenylethyl 20 2, 2- 4-HO-benzyl Methyl ( (R)-a- 618 619 bisphenylethyl methylbenzyl) amino 20 2, 2- 4-HO-benzyl Methyl Benzylamino 604 605 bisphenylethyl 21 2, 2- 4-HO-benzyl Methyl (4-Me0-620 621 bisphenylethyl phenyl) amino 211 2, 2- 4-HO-benzyl Methyl (4-Br-669 670 bisphenylethyl phenyl) amino 212 2,2- 4-HO-benzyl Methyl (4-CF3- 658 659 bisphenylethyl phenyl) amino 213 2,2- 4-HO-benzyl Methyl Pentylamino 584 585 bisphenylethyl 214 2,2- 4-HO-benzyl Methyl(2- 618 619 bisphenylethyl Phenylethyl) amino 215 2,2- 4-HO-benzyl Methyl (4-MeO- 634 635 bisphenylethyl benzyl) amino 216 2,2- 4-HO-benzyl Methyl Cyclohexylami 596 597 bisphenylethyl no 21 Phenethyl 3, 4-CI2-benzyl Methyl (4-Me-581 582 phenyl) amino 218 Phenethyl 3,4-Cl2-benzyl Methyl (4-Cl- 601 602 phenyl) amino 21 Phenethyl 3, 4-CI2-benzyl Methyl Phenylamino 566 567 220 Phenetyl 3, 4-CI2-benzyl Methyl ((R)-a-595 596 methylbenzyl) amino No R2 R4 R7 Ri-Y'Mol. Weight M+H 221 Phenethyl 3, 4-CI2-benzyl Methyl Benzylamino 581 582 22 Phenethyl 3, 4-CI2-benzyl Methyl (4-MeO-597 598 phenyl) amino 223 Phenethyl 3, 4-Cl2-benzyl Methyl (4-Br-645 646 phenyl) amino 224 Phenethyl 3, 4-CI2-benzyl Methyl (4-CF3-634 635 phenyl) amino 22 Phenethyl 3, 4-CI2-benzyl Methyl Pentylamino 561 562 226 Phenethyl 3, 4-Cl2-benzyl Methyl (2-595 596 Phenylethyl) amino 22 Phenethyl 3, 4-CI2-benzyl Methyl (4-MeO-611 612 benzyl) amino 228 Phenethyl 3, 4-CI2-benzyl Methyl Cyclohexylami 573 574 no 229 2,2- 3,4-Cl2-benzyl Methyl (4-Me- 657 658 bisphenylethyl phenyl) amino 230 2,2- 3,4-Cl2-benzyl Methyl (4-Cl- 677 678 bisphenylethyl phenyl) amino 231 2, 2- 3,4-Cl2-benzyl Methyl Phenylamino 643 644 bisphenylethyl 232 2,2- 3,4-Cl2-benzyl Methyl ((R)-α 671 672 bisphenylethyl methylbenzyl) amino 233 2, 2- 3,4-Cl2-benzyl Methyl Benzylamino 657 658 bisphenylethyl 234 2,2- 3,4-Cl2-benzyl Methyl (4-MeO- 673 674 bisphenylethyl phenyl) amino 235 2,2- 3,4-Cl2-benzyl Methyl (4-Br- 721 722 bisphenylethyl phenyl) amino 236 2, 2- 3,4-Cl2-benzyl Methyl (4-CF3- 711 712 bisphenylethyl phenyl) amino 237 2, 2- 3, 4-CI2-benzyl Methyl Pentylamino 637 638 bisphenylethyl 238 2, 2- 3, 4-CI2-benzyl Methyl (2-671 672 bisphenylethyl Phenylethyl) amino 23 2, 2- 3,4-Cl2-benzyl Methyl (4-MeO- 687 688 bisphenylethyl benzyl) amino 240 2, 2- 3, 4-CI2-benzyl Methyl Cyclohexylami 649 650 bisphenylethyl no 241 Isoamyl 4-HO-benzyl Methyl (4-Me-478 479 phenyl) amino 24 Isoamyl 4-HO-benzyl Methyl (4-Cl- 498 499 phenyl) amino 24 Isoamyl 4-HO-benzyl Methyl Phenylamino 464 465 24 Isoamyl 4-HO-benzyl Methyl ((R)-α 492 493 methylbenzyl) amino 24 Isoamyl 4-HO-benzyl Methyl Benzylamino 478 479 246 Isoamyl 4-HO-benzyl Methyl (4-MeO- 494 495 phenyl) amino 24 Isoamyl 4-HO-benzyl Methyl (4-Br-542 543 phenyl) amino No R2 R4 R7 R1-Y' Mol. Weight M+H 248 Isoamyl 4-HO-benzyl Methyl (4-CF3-532 533 phenyl) amino 249 Isoamyl 4-HO-benzyl Methyl Pentylamino 458 459 250 Isoamyl 4-HO-benzyl Methyl (2-492 493 Phenylethyl) amino 251 lsoamyl 4-HO-benzyl Methyl (4-MeO- 508 509 benzyl) amino 25 Isoamyl 4-HO-benzyl Methyl Cyclohexylami 470 471 no 253 lsoamy 4-HO-benzyl Methyl (4-Me- 554 555 phenyl) amino 25 Isoamyl 4-HO-benzyl Methyl (4-CI-574 575 phenyl) amino 255 Isoamyl 4-HO-benzyl Methyl Phenylamino 540 541 256 Isoamyl 4-HO-benzyl Methyl ((R)-a-568 569 methylbenzyl) amino 257 Isoamyl 4-HO-benzyl Methyl Benzylamino 554 555 25 Isoamyl 4-HO-benzyl Methyl (4-MeO-570 571 phenyl) amino 259 lsoamyl 4-HO-benzyl Methyl (4-Br- 619 620 phenyl) amino 26 Isoamyl 4-HO-benzyl Methyl (4-CF3-608 609 phenyl) amino 261 Isoamyl 4-HO-benzyl Methyl Pentylamino 534 535 262 lsoamyl 4-HO-benzyl Methyl (2- 568 5698 Phenylethyl) amino 263 lsoamyl 4-HO-benzyl Methyl (4-MeO- 584 585 benzyl) amino 26 Isoamyl 4-HO-benzyl Methyl Cyclohexylami 546 547 no 26'4-methylbenzyl 3, 4-CI2-benzyl Methyl (4-Me-526 527 phenyl) amino 26 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-Cl- 546 547 phenyl) amino 26 4-methylbenzyl 3, 4-Cl2-benzyl Methyl Phenylamino 512 513 268 4-methylbenzyl 3, 4-Cl2-benzyl Methyl ((R)-α- 540 541 methylbenzyl) amino 2694-methylbenzyl 3, 4-CI2-benzyl Methyl Benzylamino 526 527 27 4-methylbenzyl 3,4-Cl2-benzyl Methyl (4-MeO- 542 543 phenyl) amino 271 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-Br- 591 592 phenyl) amino 27 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-CF3-580 581 phenyl) amino 27 4-methylbenzyl 3, 4-CI2-benzyl Methyl Pentylamino 506 507 27 4-methylbenzyl 3, 4-CI2-benzyl Methyl (2-540 541 Phenylethyl) amino 275 4-methylbenzyl 3, 4-CI2-benzyl Methyl (4-MeO-556 557 benzyl) amino No R2 R4 R7 R1-Y' Mol. Weight M+H 276 4-methylbenzyl 3, 4-Cl2-benzyl Methyl Cyclohexylami 518 519 no 277 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-Me-602 603 phenyl) amino 27 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-Cl- 622 623 phenyl) amino 27 4-methylbenzyl 3, 4-Cl2-benzyl Methyl Phenylamino 588 589 28 4-methylbenzyl 3, 4-CI2-benzyl Methyl ( (R)-a- 616 617 methylbenzyl) amino 281 4-methylbenzyl 3, 4-Cl2-bnezyl Methyl Benzylamino 602 603 28 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-MeO-618 619 phenyl) amino 28 4-methylbenzyl 3, 4-CI2-benzyl Methyl (4-Br-667 668 phenyl) amino 28 4-methylbenzyl 3, 4-Cl2-benzyl Methyl (4-CFs-656 657 phenyl) amino 28 4-methylbenzyl 3, 4-Cl2-benzyl Methyl Pentylamino 582 583 28 4-methylbenzyl 3, 4-CI2-benzyl Methyl (2-616 617 Phenylethyl) a mino 287 4-methylbenzyl 3, 4-CI2-benzyl Methyl (4-MeO- 632 633 benzyl) amino 28 4-methylbenzyl 3, 4-Oz-benzyt Methyl Cyclohexylami 594 595 no 289 Naphth-1- 4-HO-benzyl Methyl (N-Cbz-3- 751 752 ylmethyl Indoleethyl) am ino 290 Naphth-1- 4-HO-benzyl Methyl (Naphth-2- 614 615 ylmethyl ylmethyl) amin o 291 Naphth-1- 4-HO-benzyl Methyl (2- 578 579 ylmethyl Phenylethyl) a mino 292 Naphth-1- 4-HO-benzyl Methyl [2-(4-MeO- 608 609 ylmethyl phenyl) ethyl] a mino 293 Naphth-1- 4-HO-benzyl Methyl (3-CF3- 632 633 ylmethyl benzyl) amino 29 Naphth-1-4-HO-benzyl Methyl (4-MeO-594 595 ylmethyl benzyl) amino 295 Naphth-1- 4-HO-benzyl Methyl (4-F- 596 597 ylmethyl phenylethyl) a mino 29 Naphth-1-4-HO-benzyl Methyl (3, 4-Cl2- 633 634 ylmethyl benzyl) amino 297 Naphth-1- 4-HO-bnezyl Methyl (2-HO- 518 519 ylmethyl ethyl) amino 29 Naphth-1- 4-HO-benzyl Methyl (3-MeO-546 547 ylmethyl propyl) amino 29 Naphth-1-4-HO-benzyl Methyl (Tetrahydrofur 558 559 ylmethyl an-2- ylmethyl) amin o No R2 R4 R7 R1-Y' Mol. Weight M+H 300 Naphth-1- 4-HO-benzyl Methyl (cyclohexylmet 570 571 ylmethyl hyl) amino 301 Naphth-1-4-HO-benzyl Propyl (N-Cbz-3-779 780 ylmethyl Indoleethyl) am ino 302 Naphth-1- 4-HO-benzyl Propyl (Naphth-2- 642 643 ylmethyl ylmethyl)amin o 303 Naphth-1- 4-HO-benzyl Propyl (2- 606 607 ylmethyl Phenylethyl) a mino 304 Naphth-1- 4-HO-benzyl Propyl [2-(4-MeO- 636 637 ylmethyl phenyl) ethyl] a mino 305 Naphth-1- 4-HO-benzyl Propyl (3-CF3-660 661 ylmethyl benzyl) amino 306 Naphth-1- 4-HO-benzyl Propyl (4-MeO- 622 623 ylmethyl benzyl) amino 307 Naphth-1- 4-HO-benzyl Propyl (4-F- 624 625 ylmethyl phenylethyl) a mino 308 Naphth-1- 4-HO-benzyl Propyl (3, 4-Cl2- 661 662 ylmethyl benzyl)amino 309 Naphth-1-4-HO-benzyl Propyl (2-HO-546 547 ylmethyl ethyl) amino 31 Naphth-1-4-HO-benzyl Propyl (3-MeO-574 575 ylmethyl propyl) amino 311 Naphth-1-4-HO-benzyl Propyl (Tetrahydrofur 586 587 ylmethyl an-2- ylmethyl) amin o 31 Naphth-1-4-HO-benzyl Propyl (cyclohexylme 598 599 ylmethyl hyl) amino 31 Naphth-1-3, 4-F2-benzyl Methyl (N-Cbz-3-771 772 ylmethyl indoleethyl) am ino 31 Naphth-1-3, 4-F2-benzyl Methyl (Naphth-2-634 635 ylmethyl ylmethyl)amin o 315 Naphth-1- 3,4-F2-benzyl Methyl (2- 598 599 ylmethyl Phenylethyl) a mino 31 Naphth-1-3, 4-F2-benzyl Methyl [2- (4-MeO- 628 629 ylmethyl phenyl) ethyl] a mino 31 Naphth-1-3, 4-F2-benzyl Methyl (3-CF3-652 653 ylmethyl benzyl) amino 31 Naphth-1-3, 4-F2-benzyl Methyl (4-MeO-614 615 ylmethyl benzyl) amino 31 Naphth-1-3, 4-F2-benzyl Methyl (4-F-616 617 ylmethyl phenylethyl) a mino 32 Naphth-1-3, 4-F2-benzyl Methyl (3, 4-Cl2- 653 654 ylmethyl benzyl) amino 321 Naphth-1-3, 4-F2-benzyl Methyl (2-HO-538 539 ylmethyl ethyl) amino No R2 R4 R7 R1-Y' Mol. Weight M+H 322 Naphth-1- 3,4-F2-benzyl Methyl (3-MeO- 566 567 ylmethyl propyl) amino 323 Naphth-1- 3,4-F2-benzyl Methyl (Tetrahydrofur 578 579 ylmethyl an-2- ylmethyl)amino o 32 Naphth-1-3, 4-F2-benzyl Methyl (cyclohexylme 590 591 ylmethyl hyl) amino 325 Naphth-1-3, 4-Fa-benzyl Propyl (N-Cbz-3-799 800 ylmethyl Indoleethyl) am ino 32 Naphth-1-3, 4-F2-benzyl Propyl (Naphth-2-662 663 ylmethyl ylmethyl) amin o 327 Naphth-1- 3,4-F2-benzyl Propyl (2- 626 627 ylmethyl Phenylethyl) a mino 32 Naphth-1-3, 4-F2-benzyl Propyl [2-(4-MeO-656 657 ylmethyl phenyl) ethyl] a mino 32 Naphth-1-3, 4-F2-benzyl Propyl (3-CF3-680 681 ylmethyl benzyl) amino 33 Naphth-1-3, 4-F2-benzyl Propyl (4-MeO-642 643 ylmethyl benzyl) amino 331 Naphth-1-3, 4-F2-benzyl Propyl (4-F-644 645 ylmethyl phenylethyl) a mino 33 Naphth-1-3, 4-F2-benzyl Propyl (3, 4-Cl2- 681 682 ylmethyl benzyl) amino 33 Naphth-1-3, 4-F2-benzyl Propyl (2-HO-566 567 ylmethyl ethyl) amino 33 Naphth-1-3, 4-F2-benzyl Propyl (3-MeO-594 595 ylmethyl propyl) amino 33 Naphth-1-3, 4-F2-benzyl Propyl (Tetrahydrofur 606 607 ylmethyl an-2- ylmethyl) amin o 33 Naphth-1-3, 4-F2-benzyl Propyl (cyclohexylme 618 619 ylmethyl hyl) amino 337 Naphth-1- 4-biphenylyl-methyl Methyl (N-Cbz-3- 811 812 ylmethyl Indoleethyl) am ino 337 Naphth-1- 4-biphenylylmethyl Methyl (Naphth-2- 674 675 ylmethyl ylmethyl) amin o 33 Naphth-1-4-biphenylylmethyl Methyl (2-638 639 ylmethyl Phenylethyl) a mino 337 Naphth-1- 4-biphenylylmethyl Methyl [2-(4-MeO- 668 669 ylmethyl phenyl) ethyl] a mino 341 Naphth-1-4-biphenylylmethyl Methyl (3-CF3-692 693 ylmethyl benzyl) amino 34 Naphth-1-4-biphenylylmethyl Methyl (4-MeO-654 655 ylmethyl benzyl) amino No R2 R4 R7 Ri-Y'Mol. Weight M+H 343 Naphth-1- 4-biphenylylmethyl Methyl (4-F-656 657 ylmethyl phenylethyl) a mino 34 Naphth-1-4-biphenylylmethyl Methyl (3, 4-CI2- 693 694 ylmethyl benzyl) amino 345 Naphth-1- 4-biphenylmethyl Methyl (2-HO-578 579 ylmethyl ethyl) amino 34 Naphth-1-4-biphenylylmethyl Methyl (3-MeO-606 607 ylmethyl propyl) amino 34 Naphth-1-4-biphenylylmethyl Methyl (Tetrahydrofur 618 619 ylmethyl an-2- ylmethyl) amin o 34 Naphth-1- 4-biphenylylmethyl Methyl (cycloheyxlmet 630 631 ylmethyl hyl) amino 349 Naphth-1- 4-biphenylmethyl Propyl (N-Cbz-3- 839 840 ylmethyl Indoleethyl) am ino 35 Naphth-1-4-biphenylylmethyl Propyl (Naphth-2-702 703 ylmethyl ylmethyl)amin o 351 Naphth-1-4-biphenylylmethyl Propyl (2-666 667 ylmethyl Phenylethyl) a mino 35 Naphth-1-4-biphenylylmethyl Propyl [2- (4-MeO- 696 697 ylmethyl phenyl) ethyl] a mino 353 Naphth-1- 4-biphenylmethyl Propyl (3-CF3- 720 721 ylmethyl benzyl) amino 35 Naphth-1-4-biphenylylmethyl Propyl (4-MeO- 682 683 ylmethyl benzyl) amino 35 Naphth-1-4-biphenylylmethyl Propyl (4-F-684 685 ylmethyl phenylethyl) a mino 35 Naphth-1-4-biphenylylmethyl Propyl (3, 4-CI2- 721 722 ylmethyl benzyl) amino 35 Naphth-1-4-biphenylylmethyl Propyl (2-HO-606 607 ylmethyl ethyl) amino 35 Naphth-1-4-biphenylylmethyl Propyl (3-MeO-634 635 ylmethyl propyl) amino 35 Naphth-1- 4-biphenylylmethyl Propyl (Tetrahydrofur 646 647 ylmethyl an-2- ylmethyl) amin o 36 Naphth-1-4-biphenylylmethyl Propyl (cyclohexylmet 658 659 ylmethyl hyl) amino 361 Naphth-1- 3-t-Bu-4-HO-bnezyl Methyhl (N-Cbz-3- 807 808 ylmethyl Indoleethyl) am ino 36 Naphth-1-3-t-Bu-4-HO-benzy Methyl (Naphth-2-670 671 ylmethyl ylmethyl) amin o 36 Naphth-1-3-t-Bu-4-HO-benzy Methyl (2-634 635 ylmethyl Phenylethyl) a mino No R2 R4 R7 R1-Y' Mol. Weight M+H 364 Naphth-1-3-t-Bu-4-HO-benzy Methyl [2- (4-MeO- 664 665 ylmethyl phenyl) ethyl] a mino 365 Naphth-1- 3-t-Bu-4-HO-benzyl Methyl (3-CF3- 688 689 ylmethyl benzyl) amino 36 Naphth-1-3-t-Bu-4-HO-benzy Methyl (4-MeO-650 651 ylmethyl benzyl) amino 367 Naphth-1- 3-t-Bu-4-HO-benzyl Methyl Methyl (4-F- 652 653 ylmethyl phenylethyl) a mino 36 Naphth-1-3-t-Bu-4-HO-benzy Methyl (3, 4-CI2- 689 690 ylmethyl benzyl) amino 369 Naphth-1- 3-t-Bu-4-HO-benzyl Methyl (2-HO- 574 575 ylmethyl ethyl) amino 37 Naphth-1-3-t-Bu-4-HO-benz Methyl (3-MeO-602 603 ylmethyl propyl) amino 371 Naphth-1- 3-t-Bu-4-HO-benzyl Methyl (Tetrahydrofur 614 615 ylmethyl an-2- ylmethyl) amin o 37 Naphth-1-3-t-Bu-4-HO-benzy Methyl (cyclohexylme 626 627 ylmethyl hyl) amino 37 Naphth-1-3-t-Bu-4-HO-benz Propyl (N-Cbz-3-835 836 ylmethyl Indoleethyl) am ino 37 Naphth-1-3-t-Bu-4-HO-benzy Propyl (Naphth-2-698 699 ylmethyl ylmethyl)amin 0 37 Naphth-1-3-t-Bu-4-HO-benzy Propyl (2-662 663 ylmethyl Phenylethyl) a mino 37 Naphth-1-3-t-Bu-4-HO-benzy Propyl [2- (4-MeO- 692 693 ylmethyl phenyl) ethyl] a mino 37 Naphth-1-3-t-Bu-4-HO-benzy Propyl (3-CFs-716 717 ylmethyl benzyl) amino 378 Naphth-1- 3-t-Bu-4-HO-benzyl Propyl (4-MeO- 678 679 ylmethyl benzyl) amino 37 Naphth-1-'3-t-Bu-4-HO-benzy Propyl (4-F-680 681 ylmethyl phenylethyl) a mino 38 Naphth-1-3-t-Bu-4-HO-benzy Propyl (3, 4-Cl2- 717 718 ylmethyl benzyl) amino 381 Naphth-1-3-t-Bu-4-HO-benzy Propyl (2-HO-602 603 ylmethyl ethyl) amino 38 Naphth-1-3-t-Bu-4-HO-benzy Propyl (3-MeO-630 631 ylmethyl propyl) amino 38 Naphth-1-3-t-Bu-4-HO-benzy Propyl (Tetrahydrofur 642 643 ylmethyl an-2- ylmethyl)amini o 38 Naphth-1- 3-t-Bu-4-HO-benzyl Propyl (cyclohexylme 654 655 ylmethyl hyl) amino 385 4-Methoxybenzyl OCH3 5-F-benzyl OCH3 470 471 386 Naphthyl-1-4-HO-benzyl Styrylmethyl OCH3 591 592 ylmethyl No R2 R4 R7 R1-Y' Mol. Weight M+H Naphthyl-1- 4-NO2-benzyl styrylmethyl OCH3 620 621 ylmethyl 388 Naphthyl-1-3, 4-F2-benzyl Styrylmethyl OCH3 611 612 ylmethyl 389 Naphthyl-1- 4-cl-benzyl Strylmethyl OCH3 609 610 ylmethyl 390 Naphthyl-1- 4-Phenyl-benzyl Styrylmethyl OCH3 651 652 ylmethyl 391 Naphthyl-1-3-t-Bu-4-HO-benzyl Styrylmethyl OCH3 647 648 ylmethyl 392 Naphthyl-1- 4-Methyl-benzyl Strylmethyl OCH3 589 590 ylmethyl 393 Naphthyl-1-Cyclohexylmethyl Styrylmethyl OCH3 581 582 ylmethyl 394 Naphthyl-1-4_F-benzyl Styrylmethyl OCHs 593 594 ylmethyl 395 Naphthyl-1-2-CI-benzyl Styrylmethyl OCH3 609 610 ylmethyl 396 Naphthyl-1-3, 4-Cl2-benzyl Styrylmethyl OCH3 644 645 methyl 397 Naphthyl-1-Naphthyl-1-ylmethyl Styrylmethyl OCH3 625 626 ylmethyl 398 3,4-Cl2-benzyl 4-HO-benzyl Styrylmethyl OCH3 610 611 399 3, 4-Cl2-benzyl 4-NO2-benzyl Styrylmethyl OCH3 639 640 400 3, 4-CI2-benzyl 3, 4-F2-benzyl Styrylmethyl OCH3 629 630 401 3, 4-Cl2-benzyl 4-CI-benzyl Styrylmethyl OCH3 628 629 402 3, 4-benzyl 4-Phenyl-benzyl Styrylmethyl OCH3 670 671 403 3, 4-CI2-benzyl 3-t-Bu-4-HO-benzyl Styrylmethyl OCH3 666 667 404 3, 4-Cl2-bnezyl 4-Methyl-benzyl Styrylmethyl OCH3 608 609 405 3, 4-Cl2-benzyl Cyclohexylmethyl Styrylmethyl OCH3 600 601 406 3, 4-Cl2-benzyl 4-F-benzyl Styrylmethyl OCH3 611 612 407 3, 4-Cl2-benzyl 2-CI-benzyl Styrylmethyl OCH3 628 629 408 3, 4-Cl2-benzyl 3, 4-Cl2-benzyl Styrylmethyl OCH3 662 663 409 3, 4-CI2-benzyl Naphthyl-1-ylmethyl Styrylmethyl OCHs 644 645 410 Naphthyl-4-HO-benzyl 2, 6-CI2-benzyl OCH3 634 635 ylmethyl 411 Naphthyl-1- 4-NO2-benzyl 2, 6-Cl2-benzyl OCH3 663 664 ylmethyl 412 Naphthyl-1- 3,4-F2-benzyl 2, 6-Cl2-benzyl OCH3 654 655 ylmethyl 413 Naphthyl-1- 4-Cl-benzyl 2, 6-Cl2-benzyl OCH3 652 653 ylmethyl 414 Naphthyl-1- 4-Phenyl-benzyl 2, 6-Cl2-benzyl OCH3 694 695 ylmethyl 415 Naphthyl-1-3-t-Bu-4-HO-benzyl 2, 6-CI2-benzyl OCH3 690 691 ylmethyl 416 Naphthyl-1- 4-Methyl-benzyl 2, 6-Cl2-benzyl OCH3 632 633 ylmethyl 417 Naphthyl-1- Cyclohexylmethyl 2, 6-CI2-benzyl OCH3 624 625 ylmethyl 418 Naphthyl-4-F-benzyl 2, 6-CI2-benzyl OCH3 636 637 ylmethyl 419 Naphthyl-1-2-CI-benzyl 2, 6-Cl2-benzyl OCH3 652 653 ylmethyl No R2 R4 R7 RrY'Mol. Weight M+H 420 Naphthyl-1-3, 4-CI2-benzyl 2, 6-CI2-benzyl OCH3 686 687 ylmethyl 421 Naphthyl-1-Naphthyl-1-ylmethyl2, 6-CI2-benzyl OCHs 668 669 ylmethyl 422 3, 4-CI2-benzyl 4-HO-benzyl 2, 6-CI2-benzyl OCHs 652 653 423 3, 4-CI2-benzyl 4-NO2-benzyl 2, 6-CI2-benzyl OCH3 681 682 424 3, 4-CI2-benzyl 3, 4-F2-benzyl 2, 6-Clz-benzyl OCHs 672 673 425 3, 4-CI2-benzyl 4-CI-benzyl 2, 6-CI2-benzyl OCH3 671 672 426 3, 4-CI2-benzyl 4-Phenyl-benzyl 2, 6-CI2-benzyl OCH3 712 713 427 3, 4-CI2-benzyl 3-t-Bu-4-HO-benzyl 2, 6-CI2-benzyl OCH3 708 709 428 3, 4-CI2-benzyl 4-Methyl-benzyl 2, 6-CI2-benzyl OCH3 650 651 429 3, 4-CI2-benzyl Cyclohexylmethyl 2, 6-CI2-benzyl OCHs 642 643 430 3, 4-Cla-benzyl 4-F-benzyl 2, 6-CI2-benzyl OCH3 654 655 431 3, 4-CI2-benzyl 2-CI-benzyl 2, 6-CI2-benzyl OCH3 671 672 432 3, 4-CI2-benzyl 3, 4-CI2-benzyl 2, 6-CI2-benzyl OCH3 705 706 433 3, 4-CI2-benzyl Naphthyl-1-ylmethy 2, 6-CI2-benzyl OCH3 686 687 434 2-Piperidin-1-yl- (S)-4-HO-benzyl Methyl Benzylamino 535 536 ethyl 435 3, 4-CI2-benzyl (S)-4-HO-benzyl Methyl 2-Piperidin-1-604 605 yl-ethylamino 2- (1-Methyl- 436 3, 4-CI2-benzyl (S)-4-HO-benzyl Methyl pyrrolidin-2-yl)-604 605 ethylamin 437 3-Pyridylmethyl (S)-4-HO-benzyl Methyl 3, 4-CI2-583 584 benzylamino 2-Morpholin-4-yl-3, 4-CI2- ethyl benzylamino 3- 439 3, 4-CI2-benzyl (S)-4-HO-benzyl Methyl Pyridylmethyla 583 584 mino 440 3, 4-CI2-benzyl (S)-4-HO-benzyl Methyl 2-Morpholin-4-606 607 yl-ethylamino 441 Naphthyl-l-4-HO-benzyl Methyl 3-imidazol-1-yi-582 583 ylmethyl propylamino Naphthyl-1-4- 442 yimethyi 4-HO-benzyl Methyl ylamino 593 594 ylamino Naphthyl-1-3- 443 yimethyi 4-HO-benzyl Methyl Pyridylmethyla 565 566 mino Naphthyl-l-2- (3- 444 ylmethyl 4-HO-benzyl Methyl Pyridylethyl) am 579 580 ino Naphthyl-l-4- 445 ylmethyl 4-HO-benzyl Methyl Pyridylmethyla 565 566 mino 446 Naphthyl-1-4-HO-benzyl Methyl Benzyloxycarb 622 623 ylmethyl onylamino 447 yimethyl 4-HO-benzyl Methyl 4-F-582 583 ylmethyl benzylamino 448 Naphthyl-1-4-HO-benzyl Methyl 4-CO2H-608 609 ylmethyl benzylamino 449 Naphthyl-l-4-HO-benzyl Methyl 4-CF3-632 633 ylmethyl benzylamino No _ R4 R7 Rs-Y'Mol. Weight M+H Naphthyl-1- (S)-alpha- 450 yimethyi 4-HO-benzyl Methyl methylbenzyla 578 579 mino Naphthyl-1- (R)-alpha- 451 4-HO-benzyl Methyl methylbenzyla 578 579 . ylmethyl mino mins 452 ylmethyi 4-HO-benzyl Methyl benzyWamino 582 583 yamethyl-1-2, 3o Naphthyl-l-2, 3-. 453 yamethyl 4-HO-benzyl Methyl Dimethoxybenz 624 625 amino 454 Naphthyl-1-4-HO-benzyl Methyl mino 513 514 ylmethyl mino 455 Naphthyl-l-4-HO-benzyl Methyl Phenylhydrazin 565 566 ylmethyl o Naphthyl-1- 456 ymethyl 4-HO-benzyl Methyl Aminobenzyla 579 580 mino (S, S) {2- [ (2- hydroxy-1- hydroxy-1- 457 ylmethyl 4-HO-benzyl Methyl phenyl-ethyl)-693 694 methyl- carbamoyl]- ethyl}-amino [4- (1, 3-dioxo- 1, 3-dihydro- 458 Naphthyl-1-4-HO-benzyl Methyl isoindol-2-715 716 ylmethyl methyl)- cyclohexyl]- methylamino 459 Naphthyl-1-4-HO-benzyl Methyl Indan-1-5g0 591 ylmethyl ylamino 460 Naphthyl-1-4-HO-benzyl Methyl PhenylGlycine 622 623 ylmethyl 461 Naphthyl-1-4_HO-benzyl Methyl 2, 6-Fa- g00 601 ylmethyl benzylamino 462 Nypmethyl1 4-HO-benzyl Methyl benzylamino 582 583 463 Naphthyl-l-4-HO-benzyl Methyl Benzimidazol-604 605 ylmethyl 2-yl-amino 464 Naphthyl-l-4-HO-benzyl Methyl Diphenylmethyl 640 641 ylmethyl amino 465 Naphthyl-1-q-HO-benzyl Methyl Furan-2-yl-554 555 ylmethyl methylamino Naphthyl-1-4- 466 ylmethyl 4-HO-benzyl Methyl Dimethylamino 607 608 - benzylamino 467 Naphthyl-l-4-HO-benzyl Methyl Thiofuran-2 yl-584 585 ylmethyl methylamino 468 Naphthyl-1-4-HO-benzyl Methyl 4-NO2-609 610 ylmethyl benzylamino 469 Naphthyl-1-4_HO-benzyl Methyl BnO 565 566 ylmethyl 4-Methoxy- 470 naphthyl-1-4-HO-benzyl Methyl Benzylamino 594 595 ylmethyl No R2 R4 R7 Rr-Y'Mol. Weight M+H 471 Naphthyl-l-4-HO-benzyl Methyl Phenethyl 563 564 ylmethyl 472 Naphthyl-1-4_Methoxy-benzyl Methyl Benzylamino 578 579 ylmethyl 473 Naphthyl-1-HO-benzyl Methyl phCFamino 618 619 ylmethyl phenylamino 474 Naphthyl-1-q,-NOz-benzyl Methyl 4-CF3-647 648 ylmethyl phenylamino 475 4-NO2-benzyl Methyl Benzyiamino 593 594 ylmethyl 476 Benzyl Naphthyl-1-ylmethyl 4-CN-benzyl OCH3 574 575 477 methyl Naphthyl-1-ylmethyl 4-CN-benzyl OCH3 594 595 Methyl 478 4-Dimethylamino-Naphthyl-1-ylmethyl 4-CN-benzyl OCH3 617 618 benzyl 479 Phenethyl Naphthyl-1-ylmethyl4-CN-benzyl OCHs 588 589 480 methyl 4-HO-benzyl Methyl Benzylamino 565 566 methyl 481 4-Pyridylmethyl Naphthyl-1-ylmethyl Benzyl OCH3 550 551 482 pimet oxybenzyl Naphthyl-1-ylmethyl Benzyl OCHs 609 610 483 3,4-Dimethoxy- Naphthyl-1-ylmethyl Benzyl OCH3 623 624 phenethyl 484 Thiofuran-2-yl-Naphthyl-1-ylmethyl Benzyl OCH3 569 570 methyl Naphthyl-1- 485 ymethyl 3-Pyridylmethyl Methyl Benzylamino 549 550 486 Naphthyl-1-pentafluorobenzyl Methyl Benzylamino 638 639 ylmethyl 487 Naphthyl-1-3-F-4-HO-benzyl Methyl Benzylamino 582 583 ylmethyl 488 4-F-phenethyl 4-Methyl-benzyl Methyl 4 CF3-598 599 4- 489 Methoxyphenethy 4-Methyl-benzyl Methyl 4-CF3-610 611 phenylamino 490 3, 4-Dimethoxy- 4-Methyl-benzyl Methyl 4-CF3-640 641 phenethyl phenylamino 491 Naphthyl-4-Methyl-benzyl Methyl 4-CF3-616 617 ylmethyl phenylamino 492 Dimethoxybenzyl Naphthyl-1-ylmethyl 4-CN-benzyl OCH3 634 635 493 34-Dimethoxy Naphthyl-1-ylmethyl 4-CN-benzyl OCHs 648 649 phenethyl 494 4-Quinoline-1yl-4-HO-benzyl Methyl Benzylamino 565 566 methyl 495 2-Pyridylmethyl 4-Methyl-benzyl Methyl 4hCFa-567 568 phenylamino 496 3-Pyridylmethyl 4-Methyl-benzyl Methyl 4-CF3-567 568 phenylamino 497 3'4 4-Methyl-benzyl Methyl 4-CF3-626 627 Dimethoxybenzyl phenylamino 498 4-Methyl-benzyl 4-Methyl-benzyl Methyl 4-CF3-580 581 phenylamino No R2 R4 Rz Ri-Y'Mol. Weight M+H 499 Thiofuran-2-yl-q. _Methyl-benzyl Methyl 4-CF3-572 573 methyl phenylamino 500 4-CF3-benzyl 4-Methyl-benzyl Methyl 4-CF3-634 635 phenylamino 501 2, 6-F2-benzyl 4-Methyl-benzyl Methyl 4-CF3-602 603 phenylamino 502 4-F-benzyl 4-Methyl-benzyl Methyl 4 CF3-584 585 phenylamino 503 Thiofuran-2-yl-4-Methyl-benzyl Methyl 4-CF3-586 587 ethyl phenylamino 504 3, 4-CI2-benzyl 4-Methyl-benzyl Methyl 4-CF3-634 635 phenylamino 505 4-C02H-Benzyl 4-HO-benzyl Methyl Benzylamino 558 559 506 Naphthyl-1-3_t-Bu-4-HO-benzyl Methyl Benzylamino 620 621 methyl 507 Naphthyl-l-3, 4- (OH) 2-benzyl Methyl Benzylamino 580 581 ylmethyl' 508 2-F-benzyl 4-HO-benzyl Methyl Benzylamino 532 533 509 3-F-benzyl 4-HO-benzyl Methyl Benzylamino 532 533 510 4-F-benzyl 4-HO-benzyl Methyl Benzylamino 532 533 511 2, 4-F2-benzyl 4-HO-benzyl Methyl Benzylamino 550 551 512 2, 6-F2-benzyl 4-HO-benzyl Methyl Benzylamino 550 551 513 2, 5-F2-benzyl 4-HO-benzyl Methyl Benzylamino 550 551 514 3-CF3-benyl 4-HO-benzyl Methyl Benzylamino 582 583 515 4-CF3-benyl 4-HO-benzyl Methyl Benzylamino 582 583 516 3, 4, 5-F3-benyl 4-HO-benzyl Methyl Benzylamino 568 569 517 2-CI-benzyl 4-HO-benzyl Methyl Benzylamino 548 549 518 3-CI-benzyl 4-HO-benzyl Methyl Benzylamino 548 549 519 2, 4-CI2-benzyl 4-HO-benzyl Methyl Benzylamino 582 583 520 (S)-Methylphenyl 4-HO-benzyl Methyl Benzylamino 528 529 521 (R)-Methylphenyl 4-HO-benzyl Methyl Benzylamino 528 529 522 4-Methyl-benzyl 4-HO-benzyl Methyl Benzylamino 528 529 523 4-Methoxybenzyl 4-HO-benzyl Methyl Benzylamino 544'545 524 pimetho, benzyl 4-HO-benzyl Methyl Benzylamino 574 575 525 Furan-2-yl-4-HO-benzyl Methyl Benzylamino 504 505 methylamino (R)- 526 Methylnaphthyl-4-HO-benzyl Methyl Benzylamino 578 579 1-ylmethyl (S)- 527 Methylnaphthyl-4-HO-benzyl Methyl Benzylamino 578 579 1-ylmethyl 528 Naphthyl-l-3-Oxy-pyridin-l-Methyl Benzylamino 565 566 ylmethyl ylmethyl 529 (R)-alpha-4-HO-benzyl Methyl Benzylamino 578 579 methylbenzyl 530 Naphthyl-2-44-HO-benzyl Methyl Benzylamino 564 565 ylmethyl 531 4'F-naphthyl-4-HO-benzyl Methyl Benzylamino 582 583 ylmethyl 532 2-Methoxybenzyl 4-HO-benzyl Methyl Benzylamino 544 545 533 4-CI-benzyl 4-HO-benzyl Methyl Benzylamino 548 549 No R2 R4 Rz Ri-Y'Mol. Weight M+H 534 3, 4-CI2-benzyl 4-HO-benzyl Methyl Benzylamino 582 583 535 2-CF30benzyl 4-HO-benzyl Methyl Benzylamino 598 599 536 2-CF3Sbenzyl 4-HO-benzyl Methyl Benzylamino 614 615 537 2-CF3benzyl 4-HO-benzyl Methyl Benzylamino 582 583 538 5-Quinoline-lyl-4-HO-benzyl Methyl Benzylamino 565 566 methyl 539 methyl 3-t-Bu-4-HO-benzy | Methyl Benzylamino 621 622 methyl 540 8-Quinoline-1yl-4-NO2-benzyl Methyl Benzylamino 594 595 methyl 54'I $Qn°line-1yl- (1N-Pyrrol-2-yl)-Methyl Benzylamino 538 539 methyl methyl Naphthyl-1-4-Benzyloxy- 542 ylmethyl carbonylaminobenz Methyl Benzylamino 697 698 YI 543 2, 3-CI2-benzyl 4-HO-benzyl Methyl Benzylamino 582 583 544 Pentafluorobenzy 4-HO-benzyl Methyl Benzylamino 604 605 545 Benzyl 4-HO-benzyl Methyl Benzylamino 514 515 546 Quinoxaline-5yl-4-HO-benzyl Methyl Benzylamino 566 567 methyl 547 $-Quinoline-1yl-3-Pyridylmethyl Methyl Benzylamino 550 551 methyl 548 methyl Pentafluorobenzyl Methyl Benzylamino 639 640 methyl Naphthyl-1-4_HO-benzyl Methyl Benzylamino (th 580 581 ylmethyl iourea) 550 Naphthyl-1-4-Amino-benzyl Methyl Benzylamino 563 564 ylmethyl 551 3, 4, 5-tri-4-Amino-benzyl Methyl Benzylamino 603 604 Methoxybenzyl 552 Naphthyl-1-4-Pyridylmethyl Methyl Benzylamino 549 550 ylmethyl 553 Naphthyl-1- (R) 4-HO-phenyl Methyl Benzylamino 550 551 ylmethyl 554 2-HO-3-Methoxy-4-HO-benzyl Methyl Benzylamino 560 561 benzyl 555 Naphthyl-l-3-Nitro-4-HO-Methyl Benzylamino 609 610 ylmethyl benzyl Naphthyl-l-4-CO2H-CH20- ylmethyl benzyl 557 Naphthyl-1-1-Naphtoylamino-Methyl Benzylamino 641 642 ylmethyl methyl 558 Naphthyl-1-4_Oxy-pyridylmethyl Methyl Benzylamino 565 566 ylmethyl 559 4-F-alpha-4-HO-benzyl Methyl Benzylamino 546 547 methylbenzyl 560 ylmettyl Benzoylaminoethyl Methyl Benzylamino 605 606 methyl 561 8-Quinoline-1yl-3,_OH) z-benzyl Methyl Benzylamino 581 582 ethyl 4-N, N- 562 Dimethylamino-4-HO-benzyl Methyl Benzylamino 557 558 benzyl No Ra R4 Rz Ri-Y'Mol. Weight M+H 563 Naphthyl-1- (R) 4-F-benzyl Methyl Benzylamino 609 610 ylmethyl Naphthyl-1- 564 ylmethyl 4-HO-benzyl Methyl Chloroethylami 536 537 nu 565 NaphthZI-1-4-HO-phenethyl Methyl Benzylamino 578 579 ylmethyl 566 4-F-benzyl 3-F, 4-HO-benzyl Methyl Benzylamino 550 551 567 2, 4-F2-benzyl 3-F, 4-HO-benzyl Methyl Benzylamino 568 569 568 3-CF3benzyl (R) 4-HO-phenyl Methyl Benzylamino 568 569 (S)- 569 Methyfnaphthyf- (R) 4-HO-phenyl Methyl Benzylamino 514 515 1-ylmethyl (R)- 570 Methylnaphthyl- (R) 4-HO-phenyl Methyl Benzylamino 514 515 1-ylmethyl 571 2, 3, 6-F3-benzyl (R) 4-HO-phenyl Methyl Benzylamino 554 555 572 3-F-benzyl (R) 4-HO-phenyl Methyl Benzylamino 518 519 573 4-CI-benzyl (R) 4-HO-phenyl Methyl Benzylamino 534 535 574 3-CI-benzyl (R) 4-HO-phenyl Methyl Benzylamino 534 535 575 2-CI-benzyl (R) 4-HO-phenyl Methyl Benzylamino 534 535 576 3, 4-CI2-benzyl (R) 4-HO-phenyl Methyl Benzylamino 568 569 577 3-CF30-benzyl (R) 4-HO-phenyl Methyl Benzylamino 584 585 578 4-F-benzyl (R) 4-HO-phenyl Methyl Benzylamino 518 519 579 2, 4-F2-benzyl (R) 4-HO-phenyl Methyl Benzylamino 536 537 3-(2-Chloro- 580 ethyl)-ureido]-4-HO-benzyl Methyl Benzylamino 634 635 benzyl 581 3-Aminobenzyl 4-HO-benzyl Methyl Benzylamino 529 530 3-N- 582 Methylaminobenz 4-HO-benzyl Methyl Benzylamino 543 544 YI 3-N, N- 583 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 557 558 nzyl IH- 584 Benzoimidazol-4-4-HO-benzyl Methyl Benzylamino 554 555 ylmethyl 585 2-HO-benzyl 4-HO-benzyl Methyl Benzylamino 530 531 586 2-Pyridylmethyl 4-HO-benzyl Methyl Benzylamino 515 516 587 4-Pyridylmethyl 4-HO-benzyl Methyl Benzylamino 515 516 588 8-quinolin-2-4-HO-benzyl Methyl Benzyfamino 565 566 ylmethyl 589 8-Benzofuran-4-4-HO-benzyl Methyl Benzylamino 554 555 ylmethyl 590 Naphthyl-1-4-HO-phenyl Methyl Benzylamino 550 551 methyl 591 4-F-benzyl 4-HO-phenyl Methyl Benzylamino 518 519 592 2, 4-F2-benzyl 4-HO-phenyl Methyl Benzylamino 536 537 593 (R)-Toluylmethyl 4-HO-benzyl Methyl Benzylamino 542 543 594 (S)-Toluylmethyl 4-HO-benzyl Methyl Benzylamino 542 543 1, 2, 3, 4- 595 tetrahydro-4-HO-benzyl Methyl Benzylamino 554 555 naphthalen-2-yi No Rz R4 R7 Ri-Y'Mol. Weight M+H 596 NaphthZI-1-Dimethoxybenzyl Methyl Benzylamino 608 609 Y Y Y 597 2-Dimethylamino-q, _HO-benzyl Methyl Benzylamino 575 576 6-F-benzyl 2- 598 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 557 558 nzyl 599 Naphthyl-1-4-CN-benzyl Methyl Benzylamino 573 574 ylmethyl 600 4-F-2-CF3-benzyl 4-HO-benzyl Methyl Benzylamino 599 600 4-CI-2- 601 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 591 592 nzyl 3-N, N- 602 Ethylmethyliamin 4-HO-benzyl Methyl Benzylamino 571 572 o-benzyl 3- 603 Diethylaminoben 4-HO-benzyl Methyl Benzylamino 585 586 zyl 4-CI-3- 604 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 591 592 nzyl 4-F-2- 605 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 575 576 nzyl 3, 5- (CH3) 2-2- 606 Dimethylamino-4-HO-benzyl Methyl Benzylamino 585 586 benzyl 3- (CHs)-2- 607 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 571 572 nzyl 6- (C Ha)-2- 608 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 571 572 nzyl 3, 4-F2-2- 609 Dimethylaminobe 4-HO-benzyl Methyl Benzylamino 593 594 benzyl TABLE 2B THE [4, 4, 0]REVERSE TURN MIMETICS LIBRARY No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weo ^ OH Zizi o N 0 H'C CH Y 802 -" 480 481 807 cNJ 430 431 430 431 r o U NYO HC CH3 N 0 HC HaC CH3 - CH, 0 H CCH, 0) 'Y Y 803 HaW NiNN 430 431 c 0 HOH 808 N N 448 449 sCH3 A a 0 su cri3 \ N O H3C CH3 804 y 416 417 NYO H3C CH3 H3C"N. 417 N O H3C CH3 a wNi N'li 9 Ao 809 9 416 417 o Ii3C^CH3 1 y-N 9) Os OH N\/O H3CY CH3 0 cl, 805 H3CuNoNtN) 464 465 oi N XNa OyN)'CH3 0, N CH N 3 w 810 /N, O A e /C CH3 N No¢O H3CyCH3 806 0 H, C CH, /nez ° \ T 0 CH, No MOLSTRUCTURE WetAhl M S M No MOLSTRUCTURE y z zizi I F \/I F nez Xfs fi N CH3 H5 NNN 532 533 N 811 446 447 0 c, N0 ° ZON N3C 0 OH F 0 Ills a 816 H3C\NNA 518 519 s wNi N N y : 0 N/0 H3C CFi3 = H3C^CH3 812 H3C"N'N N 450 451 F ,-a F \/I F \ tS ; X'rí ; + o 0 0 NEZ zu H3C CH3 'Y HaCwNiNN/I F F N \ F 813 H3CoNX 515 516 ß WF y a 818 H3CvN@o 532 533 0''O II N 0 \ c OH F 0 F p N \ Ny 0 819 H3NN N 532 533 _ C O W _ _' zon NYO I F F H3C") CH3 No MOLSTRUCTURE Welght M g (M No MOLSTRUCTURE wea ;" M S (M F F F I F F \ F NO \ I N0 \ I 820 HCNl"'T"'*"N 550 551 HC, NNN 825 Y 617 618 o \ o CH, N F F 0-)-0 N O 821 H3C, NN N 518 519 II N/\ O 0 N0 PO CH3 826 H oNs)"oNa 542 543 0 OH O N N F o. + _ 822 NN 0 534 535 i i c w w N/0 'Y 827 H3C"N"NN 492 493 o N N C CHg 823 ? N 548 549 r', ilN F N NYO 823 T"828. ! J 478 479 f 0 F N 1 II- N/0 NIN 824 HC, 552 553 o- N0 829 NC"N'INN 526 527 o 0 zizi we No MOLSTRUCTURE We s I I N J N Ci13 830 H3CuNoN) N) 492 493 835 OANq X 508 509 p N 0 N O 1 0 'on 0 CIi3 \I \I \I \I A A 'Y 831 FC' N'IN N 492 493 -a N N u C3 0 CF N 0 832 H3C, N'IN N 510 511 N N 837 577 578 832 H3CoNX 510 511 837 H35Nt ; go 577 578 0 N wC 00 II N/O 'Y 478 479 I, _ A _. bH2 H C N ^ 468 469 a wNi _N N", a 0 OH 0 OyOH XN gJNm H3C CH3 dz 834 N 494 495 I X Ueril KE ß T g s 0 OH I N I zon 'Y i F 839 H3C, N'N N 516 517 843 N, NxO 484 485 zanzi N 0 UN3 N i0 . f N 840 H, C., N"N"T'O'*'N 482 483 844 498 499 N N 0 O I N 0 1 0 0 oh ou ICH 3 . N/O I 't w B] 84s C 3 5Û2 03 N 0 N-O N O 0 HaC/I H3C . _ F/I F F N O \ H'CN'NN 842 HsW NiNN 468 469 846 567 568 N 0 0 CH3 oi o CHEZ TOC S z OH JL 0 N/0 'N n 847 N3 r NJ'r.,, iNO 508 509 85 HaWNi N 458 459 N 0 No V Nlillo con3 HIC XI N/O N/O 848 H3GNN < 458 459 853 H'N'N °, 476 477 No V No V Cl, CH3 S, CH, 0 0 849 444 445 y H3C NN N 0 854 H3C, N N-fl""N 0 444 445 0 N, c c 1 /CH3 No O OH O O 492 493 . No po O I , 855 0 460 461 I N n CH3 N 851 H'°NN 4 8 459 N 0 cl, cl3 CH, No MOLSTRUCTURE WeiBht M S M No MOLSTRUCTURE y ZIZI C3 Cl, 'f r'CH3 86 H3°wNN NJ 444 445 os 856 ° ! 474 475 cH, I N O cH, N Y or ce Ou tu non N"il- \ I'I N/\0 X tf'"3s 857 HaCs g 478 479 X f H CA II N V \ CHa N ------------------ . nez 0 i N O H3CN N ^N p/ \ I CHs 858 543 544 Ne CH3 c N 863 H3C N N N 444 445 0''O lf N v 0 N f Oh 0 H2 CH, oh 0 0 CH, 859 494 495 g64 444 445 N' -O Y C 'N chuta \ I H'C CH, y 0 No MOLSTRUCTURE Weight M+H (M No MOLSTRUCTURE Weight S) S) we9nt 1 : CH c N 0 N r-CHa r'CHa 865 H3CuNX 462 463 [C"NIN N _ sCH3 A 1 CH, N 0-) 10 CH N1 0 CH3 OH CF6 zu OH 866 H3CsN) 430 431 jj O u po o N ° ß 871 Hc'NsN)"N) 558 559 N O \ 0 OH0 CH3 0 cl, CH, N'v'CH3 i c, O N,, ,. I \ o o i 867 N'IN 0 446 447 y cH3 Nx 872 H3cvNa 508 509 UT3 N o 0 CH, CH, cl3 CH \ I O N O I/ N g H3 873 9 494 495 1 y :. 0-IN IN 0z i 'T Y" 868 ° N : lu 460 461 ccH,' o cN CH3 O , OH N I i "874 "N 542 543 -' ? Y----""- /II N O 0 c N/O 869 H3CI 464 465 Zozo 0 O No MOLSTRUCTURE Weight S) No MOLSTRUCTURE S) Weight CF CF6 ob po 875 H3CoNx 508 509 881 H3CoNx 528 529 ° wu CL, CL3 CH3 I N O I/N/O/ 1 0 876 NYN 508 509 ° 593 594 fui 0 = a O''0 H, C I-) . Y 877 N : r'N 526 527 SucH ° o N 0 po Non W I o a II N0 po o I/C^Na 878"°N,/"N 494 495 lr N° o 0 'Y , J Y'"° N N 0 0 N\,", Q 879 X AN O 510 511 _ v, 0"CFl N 0 885 N'. N 446 447 N YN it 0 880 524 525 N O I H3 FC,'N soD 0 L, OH Y) Y No MOLSTRUCTURE Wet hl MOLSTRUCTURE 886 I o. c w I o. c N/O J N/0 w w r Y HNCI IN . C, // oCH3 ", a N/O N/O 'Y 887 ° 3 \ 887 N rN 464 465 892 531 532 0 SCH, N /O''O OCHa Np Hz ou 888 N NY 432 433 i N 0 O O p = N CH3 893 H, Ns, l", zN) 558 559 0 OHO o ? ° fih --Cr N0 0-\ 889 ° 447 448 !) f\ ß 894 H3CN, 448 0 1 N/0/ MY nu 0 0 ° Y N N 0--\ - I CH3 CH3 I 890 °,,, N 462 463 o OH {% N 0/ N 0 N s wNi N zu 0-ICH, HIC ( No MOLSTRUCTURE Weloght S) No MOLSTRUCTURE Weig'hi S) 0 OH /O O OHO \ I O O N/0/O N,,, \ p . Y I 98 HaCs 542 543 901 Nz s 510 511 N CH, T yin CH, i N/O/N CH3 N 897 N3C N'1 5û8 509 902 ° ( 524 525 N O /N O 1 O OH Oui CH / 0 o I \ N O 898 < 508 5û9 X 3 X fui C N 0 C3 - 0 hic N\/O/ If N/0/ H'CN'NN 899 N N 526 527 sCH3 904 593 594 0 0 O-1 N NYO 0 H2 Y-o'Y 900 W ç G 495 oc N1- 0 No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Oh O O N i. 0 911 N 905 H oNt"vzNA 544 545 WNA o-°"' N' 0 1 /S CH3 nu0 CH3 0 912 H30, NIN-r"-N 480 481 N % N CH3 N'N 1 494 495 0 o-X CH3 OVOHo 3 CH3 0 OH 1 CH3 N -CH 1 o . N o 913 N G 496 497 907 . NI rN 480 481 CH3 NX LH3 N cl3 0 _ H3C"-CH3 nô / N H C N 908 3 N tNX 528 529 N cru N 914 NIN 510 511 o 0 914 11 510 511 N OH W I I C)) Nyo 0 909 H3C'N"') 494 495 k 0", cl3 0 YNM CF 915 H3C'N"'N 514 515 AYo oO s- zon N"O s I 910 H3C'N'NIr-N 494 495 Y.- omol CH zizi H3C') iC'N'N N 916 579 580 N zu No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight Ny 0 Ny 0 ho i3 >go lf 917 HaW NiN. N 464 465 921 9'464 465 o V'o O-CH3 0- CH CH3 3 1 N I \ N O 918 H 450 451 H c 922'N' 482 483 919 zu 923 o Z N 0 N\ /O I \ I/ 919 HaNi'" Cl, 0 oh O N o 1 CH3 O OH O . f N I \ 920 N'"foo"N 464 465 924 N N 466 467 N "ICH, Cl3 N CH3 N CH 3 No MOLSTRUCTURE Weloght S) No MOLSTRUCTURE Weig'ht S) S1 5) N CH, N N 929 H3C, NN N 430 431 925 480 481 0 4 X O CHa t O CH3 O OH CHa /\ cl3 NYO 0 ).) 930 uc, i) J 416 417 NIN y'I 926 H3NNN 484 485 No 0 N 0 H3C CH3 . Ns"°) Cl, \ I \ CHa N/O I 931 H3NN N 464 465 N o o HC"N y- N NY 0 N 927 549 550 o zack 1 c3 N O 932 H3C NIN N 430 431 Oh CL, "Nl N o CH 3 928 F6c N 480 481 N I cHa O < XrD 933 H3CI N'N 430 431 N 0 o NC") CH3 No MOLSTRUCTURE Weight No MOLSTRUCTURE Weight 5, S CH, xi CH3 CH3 N O N O 934 H C tN V 448 449 n 0 0 II N il N ci, zu SCH3 \ / \ C \ CFia HaW iN 935 H., NNN 416 417 N N 939 515 516 o 0 0 z N Cl, t 0 k 'N C 3 936 N'IN 0 431 432 0 1 y N 0 CH3 N'\ J,,, N N $ 940 H, C'"N 504 505 u N CH, I N0 \ a NYO N 937 °" j/ 446 447 941 H3°N'N 454 455 O C CIi3 O ni Cl, 0 N 0 942 440 441 H wNi N 0 _ I-IC^CI-t3 No MOLSTRUCTURE yyeght M+H (M No MOLSTRUCTURE weight S) ß to O Y OH ° 9 43 {HC> P | 488 489 i | | r N { { we 0 OH 0 Nu 0 0 i 943"3°NNN 488 489 N 0 948 N p 455 456 1 N'IN 0 c3 N N O N 0 944 H3CN NN N 454 455 o con3 cl, N ci N o N Ha 345 HaC i X 454 455 949 l 470 4 1 N01-11 0 y-N 0 0 = ,/O O OH 0 OH N/O / 946 HC NN N 472 473 N T'Y 950 H3C NN N"En 474 475 son N 0 \ d N¢O in1 t 440 441 N 0 CH3 S N O | k 1 v X R U C T U R E X D No MOLSTRUCTURE yye9h M+HM No MOLSTRUCTURE ^^. M+H M Weight N O N/O "3WNNN 956 H'CNI°N Y N 554 555 0 _ : 0 MHz MHZ 0, zizi cl, y tiz OH 957 HC, NN)",-, N 554 555 N. Zon 0 N CH3 0 H3C 952 H3C N 604 605 N \/I NI 958 H'N'NN 572 573 zon \ I II 0 0 S, CH, Y I \ SCH9 953 554 555 N zon o /cr 'N CH3 959 H3C'N'NN 540 541 w I II N o/ o N O ' o w ciw-o/N zon g60 N o N'556 557 a L ; H 3 N 955 H3C, N'NrNe 588 589 N O \ 0 : I// No MOLSTRUCTURE weight No MOLSTRUCTURE Weight 0-1 1 YN CH, N 0 1 N 966 y 464 465 T"T A N O N O 0 can O L'OH H3C CH3 N 0 N 0 1 N/O I/ N0. fi 962 H'°NNN 574 575 96 H3CNNN CH 512 513 zon zon 0 y 0 I i H3CININ N NYO i0/= 987 3 uNv Hs õ12 13 o \ erz N O 0 CH3 CH ou \ po o 0 N I O/ N 983 | oo so 839 840 988 X CH 478 479 964 HC N 0 H3C") CL, __ xi N 0 N'N CF 478 479 970 N N CH3 0 a XV LXD CH3 CH3 No MOLSTRUCTURE WMeilhl M+H (M No MOLSTRUCTURE w lei hi M+H (M Ao 0 Y° ? W W 1 y 971 H3CsNX H3 464 465 N N CH3 - N 975 563 564 ouzo c3 N Cl 3 N O CH3 Cl, O N. I I oH 1v'OH 0. fY t'o/ po 1 y N 4 976 Hc A"t 582 583 Nlilo ci ci N CH3 N/-o ci 0''N'N 977 H3N N 532 533 cri a ''AN L, o 'CH, Cl CH3 0 OH Ou nez zu 978 w y cl 518 519 N O I \"'o I CI /- H3C^CH' 974 H3CsN X CH3 498 499 oui 0 cri L I O I I X (f 0 YN C, ''O 'CI No MOLSTRUCTURE y, ee ;,, M+H (M No MOLSTRUCTURE . $ WeIAhI N0 H3Cl I N CH, WN O iacl 985 Cl < ^ KNXO 548 549 985'548 549 0 ci c I Nio 0 I N CH y cri / 0 N O CI Y 981 H3C, N-"-N 532 533 Cl ! ! !} ! ci H, C') 06 Cl, cri 0/ N 0 CI y ci 982 >¢° 3 Cl t f _ . N CI lf 0 CI \ ^ p H'N'N -N/ 0 N'.) a Y. v' I 1 0 C ! CH3 987 9 O C, 618 619 i, \ N0 y cul 983 ^ 983 N-N- 518 519')., I I CH, O 0 0 OHO 0 0 OH N 0 988 H3NNJ''--,/N 482 483 0 N/N'O O 1 Uhr 984 CH3 N 534 535 I y C3 N I \ \ I Ha CH z 989 H3NN N 432 433 po 0 =., CH, lez CH 3 No MOLSTRUCTURE Weight S) No MOLSTRUCTURE s i i CH3 990 y 990 H3\NO 0 418 419 995 "° . J 418 419 gg, J 8 419 -, a N N N N 0 O H3C CH3 ci 0,.. OH CH3 0 N\/O p 991 H3C'N,-"Nf 466 467 N,,,, lg, N/t 996 INH3Nt° 433 434 N-v \0 996 NYO XCoH3 433 434 0 \ CH, N un3 N cl3 0) 1 NYO \ I 432 433 'N 1f p 0 997 0N 447 448 1 "N0 CH3 H3C N 0 OH PH Ou N0 v N O'p f 432 433 cH3 zizi N N0 0 998 H3CI : NrNf 452 453 3 N¢O 8 H3 v 0 i (N\/4 NYO IN) I 'nez Ny0 u SCH3 No MOLSTRUCTURE Weight S) LSTRUCTURE weight cl, I c'I N/O O N/O n ci 1OO4 NNN 498 499 zon 999 II ° 517 518 No 0 = y = 0 z N 0-)-0 Oh 'cl 1005 HC"N 498 499 N O II N ) a 1000 H3C'N 548 549 H3c N' O/ \I I/ 1 cri /CI 1006 N"] N"aN 516 517 N CI 1001 HC, IN r 498 499 N o 'c 0 cl3 /I N0 \ CL 1007 NC NN N 484 485 N CI 1002 Hic N 484 485 0 3. N. C N CH3 H C^CH OH CI 0 cri '/ o 0'N 1008 N o 500 501 N N N 532 533 L ; H N N ICI 0 \ No MOLSTRUCTURE Weights) No MOLSTRUCTURE ci ce NEZ O \ 1013 . N'N N 518 519 . N : L N o N 0 = N H3C^CH3 ci 0 OH ci my 1 : ci 1014 °NNN 566 567 cl N 0 1010 N N 518 519 \3 _ N v 0 ci ce y 0 0 1015 C', NN 532 533 N cri Zon 1011 II ° 583 584 o \ i cl "I N el \ H2 N'.'NIN 532 533 cri 0 i cl ce 1011 I CH3 A ßf0 tCI 532 633 cl X l fo f Cl { /CI XI 0 CH, V Y 1017 3 uNo) N 551 552 ZON y : 0 0 I S s'CH, No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight M+H (M cl ci 1 : ci y H3C, N N0 \ I N0 \ I 1018 3 uNz) N 518 519 N 1022 II ° 618 619 II N N CHAN 0-10 ciel CL ci OH 0 N1,,. I I/CI OH 1019 Cl3 N 534 535 AN 4oH. 1 CH3 N J N 0 1023 H3C"NN 528 529 N10 I chez // CH3 CN3 N CH3 1 1020 ° ; 548 549 ce 1024 H3N''"N 478 479 ce 0 L'OH 0 0 cl3 ,-r ci CH3 I ci CH3 Cl /CI/CH3 N i0 (\ 1021 itc, N N 552 553 N 0 1025 H c N 464 465 0 H3c, IN N zu HC'"CHg No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight S) 4 to 3 A to 53 CH, CH, : CH3 Nu 0 N 1026 N>lS*° tc3 OxN) BCH II NO II NO N N /cl3 O oh Cl, 0 OH N O I 0 N1 I CH 3 1027 N 1031 480 481 0 °Y CH. 1027 N N 478 479 cH O II N a N 0 ICH CH, CH3 \/N I Ha N 'Y° 1032 TY 494 495 1032 : Lo 1028 3 NiNN 478 479 N N 0 0 OH O Hz CH3/ CH3 I /CHa \/ \ I/NO \ I X J 3 j H3C,, N N N 0 MHZ No MOLSTRUCTURE Weight M+H (M No MOLSTRUCTURE Welght S) i I c i I N 0 N O N o I \ H3GoNF ZON 1039 H3CsNz 478 479 N N ii 1034 n i 563 564 c, zon O--A, O Nu "'Y° ri oh 1040 3 N N 478 479 \ OH II N 0 ; N | H c H N 1035 H3C 528 529 X 0 N O 1041 N"r 496 497 'nez O At0 VNA 1036 3°NNN 478 479 Nez 0 CH3 y II C N 0 \ 1042 HC, N'N 464 465 Hz 0 \ ° r." 0 kA, AJ 1037 H3, N I 464 465 0 oH H3CNN 0 N °i, c cH, Otc N 1043 NN O 480 481 I CH3 N 1038 I% ClNy0N"'C 512 513 W N W O No MOLSTRUCTURE Weight No MOLSTRUCTURE Weilht M+H (M k Weight nu0 N . O N CH3 1050 H3CsN, NsI NX J3 540 541 1 1044 ° ,,, N 494 495 o N ° N O OU i N O OH o O 1 1051 51 506 507 a y chez , 1045"°No I 498 499 °H' zon O I ol/I NO \ 1052"°-NN 506 507 - NO Foc CFL 1046 563 564 NYO I 0 1053 H3CsNW 524 525 0 lof O Ici = oh po O/ O N 1047 tN4OH _ _ CHa __ 1047 HaCz SN) z > 556 557 f e N)--O 1054'6c-N N 492 493 CF tu cl) po N o N I 1048 506 507 1055 NiN/0 508 509 MY CH, w, j ! 1049 H3C\N, ; WJ 492 493 'nez 0 I H3CCFl No MOLSTRUCTURE Weight M+H (M No MOLSTRUCTURE Weilht M+H (M Weighl I Ftc Fi3'Y N 0 L. OH OH my /O OH I/ N i0 F 1057 f) 1063'''N 496 497 "3°,-"rN 5 527 T ° S CHa _ , A. 0 O/ cuti, \ N O N O F FC'N 1064 HC,--496 497 1058 0 ° 591 592 0FACS CHEZ H2 OH Ny0 F \ OH, I 1065 ITCNN N 514 515 O _N J l N 0 Ip = f ou N OJ F 1 O F N O F \ 1066 FC, N'y"N 482 483 . N 0 F 0 y 1060 HaCsN « NX 496 497 c 0 0 fez c3 t 1067 NN\CO 498 499 N OF 1061 H3CNg I, J 482 483 ) FC 11-1 CN 0 No MOLSTRUCTURE Weight M+H (M No MOLSTRUCTURE M M+H (M $) Weight / N O 1068 1--N IN y 464 465 93/X/Nt H, C CH, N 0 H, C--CH, N OF 1069 ItC'N'N N 516 517 y 1074 HC'N"N N",, cH. 512 513 0 =-a N 0 0 N y0 F etc NNN 1070 581 582 A 1075 H3CuNX CH3 478 479 0 0 OH CH3 0 ou CH3 N O I 1071 H3NwN/'..,, N CH3 528 529 N\/O dSo t 1076 H3CuNXtCH3 478 479 N O- 0- HC CH3 nu 0 9,- 1072 H, C, N C 478 479 \ N 0 o \/cH3 1077 H'NN ''cH, 496 497 ly.-o I N O- 1 SCH3 No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Mo. M+H M Weight o 1078 H3CoNNtN'CH3 464 465 H3CN'N<N"CH3 N 1082 563 564 0 c3 N cl3 N O CH3 0", CH N CF 0 N 1079 NN0 480 481 N 481 No w w cH3 N 1083"3wNNN 514 515 ) a _ 0 O 0 cl3 -r CL3 O''N'NH3 1084 H3. N'NN 500 501 No N 0 0= OH3C CH3 Cfi3 O OH // Nu0 1085 H'CNNN 548 549 \/N N0 0 N O 1081 H3C, N"Ill N""CH 498 499 ,-a ° i i 0 1 Ok. 0 "CH, O \ "CH, CN No MOLSTRUCTURE Wefght M g (M No MOLSTRUCTURE Weight S) NYO NYO 3 1087"3NNN 514 515 1092 H3N"N 534 535 N OH C\ ; 0-'=) 0 CH3 \//I \ I -room N O N/0 \ \ I 1088 H3CNN n NJ 532 533 H3CNN n NJ Non 1093 599 600 o 0 CH, 0--l-0 N 0 . CH, OU 1089 H3C'NN 500 501 A OH _ N 0 0 y 0 9 N S ß N v CH3 1094, NX J ZNA 520 521 of OHO/ N Y'nez O N\, , ol 1090 CH3 N 516 517 Con3 N CH, N6 \ I/N/0 9 1095 H3C\N X 470 471 Q lez 0 NCH3 0 CH3 1091 OA NX X 530 531 CH3 p N O razz 0 T 0 0 No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight N 9 N O 0 LJ NO LJ 1096 H, C X 456 457 H c 1100 3 N'rN488 489 II o II wiWo 0 0 \ 1 SCH3 cl 1 X I I Ny0 1097 H3C,, N'INN 504 505 N N 1101 H3CuNWo 456 457 CH3 0 cl3 /cl3 O oh 9 N O 1098 f _ VN _ o 0 1102 N 472 473 erz C3 N ICH zizi N/O 1099 HaN''"N 470 471 N CFI3 N 1099 1103 486 487 _ H3Cn _ Q N) O C Nio ZON 3 Ny 0 oh No MOLSTRUCTURE M F F UCTU E C ? S) Wetght S ( F N 0 y0 1104 H3C, N N 490 491 1108 H3C' NIN'IN CH3 530 531 O O II N II N 0 0 /I F F N/0 N0 I/ 1109 H'N'NN CH3 496 497 N N 1105 Y : 0 555 556 i 0 N H N fLJ /I-F F Ny0 NYO F 1110 HC, N. N c 496 497 N = 1106 H3NNN CH 496 497 H3c ) a CH N 0 3 O 'C 3 CL3 I a nez F 1111 H'NNN CH3 514 515 N I/l N v -O 1107 Hc N 482 483 0 N N CH, ç Us N 0 CH3 0"ICH, H3C F Nu0 1112 HC"IN 482 483 N CL, 1 0 No MOLSTRUCTURE Weight M+H (M No MOLSTRUCTURE wight S) 0 OH0 OH N 0 O OHO CH OH -o O 0 i ! 1117 H-cN" 542 543 1113 NN° 498 499 N'o tc w Ut 3 N N, r N i0 1118"3°NNN 492 493 1, Y.. ; 0 3 N 1114 r 512 513 o cH, Ha nu I N I CH3 O OH N O ° 1119 CH, N"4 X OH H3 _ N y N N O CFia \ \ i C CHj / 1115 H3NNN CH3 516 517 , N N\/O 0 N 0 ° t3 1120 H3C\EXt 526 527 F6 0 0 F , N N CH3 N10 1116 WN>o 581 582 1121 H3CsNS 492 493 A XCH3 0 0 i N 1 pi CH2 CH3 i N N'0 My 1122 H3CI NIN N 492 493 H, C- Hz No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight S) su I C N\O I \ N I O I 1123 NCIN IN N oiu 511 3 1 NI'f"."N :-FLO ° \ CN N 0''0I \ 1124 NN"'N , , o H3 0 N O H3 N ) H3 1129 HC N, N)"",, N 550 551 O/NO F Oh F 'q O N,, ,, I CH3 I F 1 1125 Non 0 494 495 ruz CH3 N/_0 F 1130 H3Cl NI N 500 501 N F c9 Ho / N H3 N 0 1126 °N 508 509 1131 'N r,"Nio 1131 H3CN NIN"N 486 487 N Y--""O F I/O L OH H C^CN o Y F F 1132"'°NNN 534 535 1127 N 512 513 F , zu b 0 No MOLSTRUCTURE weight S) No MOLSTRUCTURE Weight S) Weight NYO F 1133 H'CN'N y N w 500 501 F F F N : lu ZON 0 OH oxon nu0 F 0 F 1134 t fo CH, F i ; 4 F i yOF 1139 N""f"'N 520 521 O CE, I/ 0 CL 01) 0 'Y F 1135 518 519 Ny0 F H3CI 0 _N 0 F 585 586 S, 0 Zon N N'0 N0 y F H'NN Y N OH O F / p 1 O po 0- ! !) \ A\-° N'O/I au O N, /F H' 1137 1137 501 502 1-, y UN3 N y0 w 1142 H3CI NNrN 538 539 1f N O o 0 \CH, CH, No MOLSTRUCTURE Weight M+H (M l No MOLSTRUCTURE 9 M+SH) (M Weight HC p OH o I Ha N 0 Nu N O N/0 1143 L ; N3 1149 NN/0 540 541 MY Y H3C^CH3 N / Ns¢O A O i a I 1144"'°N'" 572 573 N o N Na "-° 1150 554 555 Cl6 nit r 0 CH3 0 OH N O my 1145 H'CN'N -N 538 539 i o NYO fez 1151 H'°N" 558 559 in NYO C O H3c, 0 CH, 0 10 1146 H3CS LS 5 38 539 H, CXN, ANA 'N N \ HZ LjL H3C") 1152 y : 623 624 /I N O ° cI H3 0), N yo l-0--lO 1147 N'. N 556 557 NO Chlz Lt OH 1. c5 0II po 0 N6, N 1153 H3C'N 508 509 1148 No No HC N N ^ 524 525 CH3 _ _ CF 'o'O \ CH3 1 No MOLSTRUCTURE Weight No MOLSTRUCTURE Wea 154 ; RC N) NX ; 408 0 459 [CC ¢ (i WN w>C°H VNa CH, CH, 0 nu0 N O 1154 N O X A 0 H3 0 II N O ce 3 S Noch3 / I N 0 1155 N3\N' N 444 445 1160 H3WNNN 444 445 O N 0 H3C CH3 0 CH3 CH3 CH3 0 OH Cl) 0 N/O NCH 1156 H3CN'Nn 492 493 o N, I : 0 1161 460 461 o I CN CON3 \ CH, 1157 H3CN'NN 458 459 N CFI3 II N° 1162 oi, N IN 474 475 ° we f. l° 0 r". Nio CH 0 OH My O ou nu0 O CI-4 Ka' 1158"aWNN"458 459 No 1163 H'CNN 478 479 N 0 H3 No MOLSTRUCTURE Mo. M+H (M No MOLSTRUCTURE ^^. M+H (M CFl /C N--o I/ N O ß Xo X 1189 % 588 5 9 o, zozo 1164 543 544 0 l N po ouzo I oh ll N ° HC) 0 ct rl-N o r en, 1165 H3NN'',./N/618 619 NI-o N10 S e 1171 H3CoNX 586 587 ou lu ici S c Ny0 1166 H3CvNt 568 569 NorO Q Nu0 \ g Le mnE Cl, 0 N o 1167". N N 554 555 0 oH zizi N etc 0 N 0 N tYo Q 1173 X CN ? t ° 570 571 Nez Y° 1168 HC, N". N 602 603 , Ck I zizi riz No MOLSTRUCTURE Weight No MOLSTRUCTURE Weight S) i I NCH, N0 CN 1 1174 ? IN 584 585 1179 H3CoN<CH3 430 431 I f /NYO 0 LH ' 0 Y 0. \ N/0 CH3 'Y "Y° Y 1180"3NCH, yg yg 588 589 No 0 0 o I 1175 tC'N'N 588 589 VNa 0 0 N 0 y N0 CN 1181 tq N"CCF6 444 445 0A0 O 0 I cru 'raj I 0 1 N0 CH, v ßNf 1182 H3CNN/Nn XCH3 444 445 1177 H3C N)/CCH3 494 495 WNS Y : /cl3 \ ni X N\CO RCH3 1183 N'--r-N-CCF 462 463 1178 HAN NC4CH3 444 445 VNS 'N O in 0 ICH3 SU CHEZ CH, No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight S) N 0 CH, y0 CH N O CH3 N 0 CH3 1184"3N N 430 431"3N" zu 1188 II 529 530 H3 N O OH CH Cl, N v Noch 3 oh 446 447 1 y 0 1189 H3NNJ"'--,/N 506 507 N) iso I N CH 3 N0 I 1186 N 460 461 1190 H3Cs, Nt X 456 457 neo Ni 0 N O 'ha zur O L'OH H3 I i oui l CH 1191 F6c,, 442 443 non 1187 H3CN, N<N CH3 464 465 WNA , o 0 H CF H3CChl3 1192 H3GuNX 490 491 ) a N y-0 ( No MOLSTRUCTURE Weight No MOLSTRUCTURE Weight S) su O O O N o ^'N 193 o N I "IN0 'N "CH, j ! 458 459 N 0 1 y zero N10 w 1194 H3CNt W) 456 457 9 a 0 N CH3 c ? C 0--N 1198 ,, 472 473 N o N N/0 O SOH 1195 NC N', N N'O 474 475 'N N \ : S, CF N0 y 1199 476 477 neo N 0 0/ 1196 . NNN 442 443 z 0 0 i 0 = wu r."3 CvN N _ N V 1200 541 542 zon CH : No MOLSTRUCTURE Weloght S) No MOLSTRUCTURE Weight S) Weight Su oh/ /Br O N I 1201 H3C sN) 592 593 1206 H. C, NIN 542 543 N O CH, CL, /ber N0 \ I NO \ I [Aa Br X, Br 1202 1207 N) Nk 561 562 CH3 zozo 0 con 0 _ Ber Y . _ CH3 _ CHg N O N Q Y' C 1203 \ 528 529 'i 2fl8 . N. NN 528 529 N y.. o CH, O oh cil r N/0 N Br 1204 VBr OaN Br _ zizi ° 1209 5 4 5 5 nui Chu _ N, O I I 1205 CNNN 542 543 ii NO I Hs N 1205 1210 558 559 i N o Cl, N 0 OH yy ? r O No MOLSTRUCTURE mn S T R O zT U R CF ber zu N0 N 0 1211 N-T,"*N 562 563 1216 H N NJ 522 523 a w iN O N N 0 0 zur ber 0 w I sr N O I 0 N 0 H, C" N-N 1217 hO N"""N 488 489 Y. N 1212 628 629 0 0 MHZ 0''0 l 1 0 CH j OH 1218 yo 488 489 o 0-a N : Ms D X 1213 " 538 539 1213 0 C3 /O 0 N O CF6 1219 itc, N'r'N ? 506 507 N"I, \ I N ! 0 0) 0 ? ° 1 1214 I/488 489 N.'N CF Z 'cl, o 5 o N..,. 0 J 0 Zon 0 nez N O J = 1215 r 474 475 li'CN'N Y'N- nez _ O I I No MOLSTRUCTURE Weght M+H (M No MOLSTRUCTURE ^^. M+H M S) S) 0 OH O ohm 'il Y-NOCI-I O N I N/O 1221 IN 0 1226''-N-'N-'558 559 A. Y CH3 N iNa y o \ N 0 N , O YN CH 1227 N 524 525 1222 °N 504 505 \ : L 504 N 0 D ici Cl, y 1228 524 525 ou N osa N O H, c 1223 H3CoNt 508 509 CH3 _ N N O r n 1229'c'N N)," N'-'-'So 510 511 CH N zu N, r NO 0 OHO/I N Y N 'N'"aN i O. N. 1224 0 573 574 123 NN0 526 527 jH2 N N cl, po yin cl neo 0 N I Ha 1225 510 511 1231 0-1-N N 540 541 N O dz T 0 No MOLSTRUCTURE we9n, M+H (M No MOLSTRUCTURE M. M+H (M Weight S \ \ CI 1232 lC, N-N 544 545 rt 1 0SL ^"'X 33 0 , CI 0 609 610 1233 1238"nu 498 499 0 H2 0 OH CN 0 ou N cul N 1234 1234 N 548 549 N O NO \ T ! 1239 N N 498 499 po N ci . ___ 1235 1239 N. N N 498 499 No N 0 A F-_ ° _ CI-l lf N ! 0 ! 0 0 ci CH, N 0 / 1236 Cl 484 485 cl Non N N/O 0 1241 N 484 485 N ° cl, No MOLSTRUCTURE No MOLSTRUCTURE 0 OH 1 OH cl p 0 N N O N1 I I ., I 1246 c'NNJ"'°N 534 535 1242 N 500 501 + :, aX I \ y 0 1s I 1 1247 H3C, N N 484 485 Na N I N 1243 514 515 o CH, IN zea CI 0 OH ou -. _.. N 0 ei 1248"3c, y ICS"470 471 /N N kJ 'o 0 N 0 1244"'°NN 518 519 °ccH, 'N "a 0 ils 1249 H3CIN N 518 519 cri N 0 I \ CN N,/ 1245 lO ° 583 58A. H3C y N 0 rS 1245 ft i 583 584 r)) jH2 1250 H3CuNX 484 485 ni Y"1 0 cru CHt3 _ No MOLSTRUCTURE No MOLSTRUCTURE Weight S) Y° v'Y° V 'Y 1251 3 uNz) N) 484 485 1256 H3CNNtNJ 504 505 via No N Chez y., 0 lao o 0 1252 H3CN'NnN 502 503 H3CN'N) N o . 1257 o 569 570 o \ o s. con ouzo Oh ON 1253 H3CuNeo 470 471 tNX 11 o o po ON CH, 1258 "" 536 537 0 OH0 N"o CH, CL, CH, /CH3 1254 NIN 0 486 487 CH CH, NYO I 1259 H3C"N 486 487 N Y"0 0 cl3 CH, N CH, CH, 1 1255 ° ", 500 501 N : Lo N0 1260 y 472 473 ' N N N 0 0 0 H, CH, No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight S) 0 oh /H O OHo 3 CH, i N CH, Y Y YY \ y 1266 N0 CH, 488 489 f X 1287 | J X X 502 5 3 WNa H3C Nt y CL3 zu 0 cl3 \ \ C N i0 HsC N I 502 503 1262 H3C, N"N"ro'*N 486 487 N N H3C N 0 : Y y CH \ CH3 N/0 My 1263 N 1268 H3C, N'NN 506 507 NYO kj H 0 0 I CH3 H3C CH3 XA N\/O 03 nu0 1264 NN504 505 N 0 N"I y= o \ 1 y 0 N \ dz CH3 CH3 Nu0 1265 H3c', 472 473 /nez CHEZ Y.,. 0 Mm. M+H (M No MOLSTRUCTURE ^^. M+H (M No MOLSTRUCTURE we9n S Weight OU / N0 Nez 1270 NKN""oNx 558 559 X u N0 0) H, 0 oh O ou oui N O N 1271 iAcsN<°< 508 509 1278 CH3Nrs 510 511 N ° S zou 0CF "Ir N O \ 1272 C, Y 0 o,, o 494 495 i ho N 1279 524 525 At0 CF 0 Oh o ou 1273 N N--'--0--C 542 543 o, N O 1280"3°N'rN° I 528 529 yin 0 O/ FC, y 1274 'CNN 7 N I/ yin o 0 0 I H, oN 1281 o N° 593 594 itc"Y 0 Y jf Y 1275 ; d3Cs/0\/l3 508 509 Win3 OH zozo N O N 0 _ A H3C) CH3 _ tN XOH _ 1276 H3C INN)""""IN 506 507 'N : o No 0 zu S, CF show No MOLSTRUCTURE weioht s) No MOLSTRUCTURE WeSght M S CH ICH N 0 0 Ns¢O (0 1283 H3CsNt) 456 457 1288 H3CvNC 474 475 C 0 inc CHUS SU , CH C N 0 0 1284. c TJ 442 443"Y° r 1289"°NNNJ 442 443 = o N ° 3cc 0 1 /cl3 0 H3C CH, 0 N/ 0 CH3 1285 H3CuNX NS 490 491 o N N 0 1290 N 457 458 0 1 cl N cl NY 0 f 0 Ok 0 r° 0 1286 H3CuN 456 457 _ VN CH3 N CH3 0 N 0 1291 0 N 472 473 '°, \N' _0 Hic" zozo /0 OH CH , CH N CH O 1287 H3CNN4bS 456 457 A 4CH N\/O 0 0 1292"°NN N 476 477 tri C H3C) CL3 O No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight zizi N Ns¢O 1 1299 H, Cs, N) ~õJ3/522 523 ; o 'kA. 1293 o 541 542 o 0--0 cit 1300 540 541 Hz 0 OU z OH ScHa oit I O \ N N, N o \ cHa 1294 1 572 573 1301 FC'N"Crc 508 509 C tCH3 inf N two _ l. nez Il Nô chez cl3 /O I C I o cH'O N 1302 N'524 525 0 cA C- lez cil 0 cit N . O CFi i 1296 y 508 509 Y\o 1303 °" 538 539 P,., No u Y. 1297 ITC'N 556 557 0 NO/I \ \ N"'N-' 1304 H'CNI'N T N 542 543 rY 0 N I O 1298 522 523 o a. o oS o o I 1305 607 608 'N zu 0 No MOLSTRUCTURE Wefght S) No MOLSTRUCTURE WeAh M+H M OH F X) OH/I F I\ o O N 0 O N' 1306 0X _ _ X Na N"'O 0 FOB CHU F F Cfi3 F F N 0 0 . Y N i0 0 1307 526 527 1313"3NNN 512 513 N 0 CH, CH, 3 Hz 0 OH N0 0 i Y'N N O O 1308 Y 512 513 0, 1314 NiN/O 528 529 CN3 N O 3C^CH3 F foc N O O 1309"°NNN 560 561 Nul 0 N 131 5 F NXo 542 543 F N : Lo OU or F I /OH O F N O 1310 H3cN NIN N 526 527 N 0 . Y 1316 546 547 0 o CH 0 Fo 1311 Rit 528 527 Zozo N Hz HIC No MOLSTRUCTURE Weioght S) No MOLSTRUCTURE we, e ;" M Weight F I \ F N _0 0 I/ Y HCNN ^N N O 0 1322 H3CNN N 526 527 + fS X ß 1317 0 ° 611 612 0 " N I oh 0 0 HZ \ OH I I /N 0 O O f 1323 nc . j 526 527 N 576 577 N O 19 F /I \ 3 if F F N 0 O 1324 H3C'N : Nr0 Nf 0 544 545 N 0 G 1319 526 527 0 Nez O SCH, 0 cl9 CL3 H3' F I I F 1325 I N 0 O o 0 1325 H3yNi, N 512 513 1320 N N 512 513 No H9C N ^ _ N . 0 n O OH H) n °Y'""' F I 1326 N'N 528 529 N0 tu 1321 J 560 561 1 II N0 I/ y z zozo No MOLSTRUCTURE MS M+H (M No MOLSTRUCTURE M0t M+H (M Welght S Wei9ht \ i fiv 1327 N 542 543 N o 0 HC, N-N 0 nez I i o oH N O _ my F H3CnCH3 H3C CH3 N 0 p' H, C N p/ 328 WCot a, ? X o 546 1 547 °'Q N v O W O = I i Ny0 F F F 1334 N'.."r : N'-"O 526 527 r6 YNz N 0 0 0z N O O p' H'WNN N oH 1329 VNSo 611 Cl) O NO F I 1335"°NNY. No z S ctX 3 W"s WSo 91 O C : ho H'C I Oh Fui 1336 544 545 1330 576 577 0 Y k N O O 1 \ I I \ SvC F N"O 1337 NN'rN"'0" ( : 512 513 N, _ 1331 HC'N 526 527 ly 0 0 z I) H, v- No MOLSTRUCTURE ^^. M+H (M No MOLSTRUCTURE ^^ M+H (M 0 0 OH F zoo N0 N'" 528 529 en N N'o 442 443 neo 0 CH, CL, i CH3 N I H 1339 542 543 N N jL Yo 1344 H3\N N 428 429 zozo OH w H3C^CHa my 1340 546 547 0 0 NYO w Y n 1345 H3°NNN1 476 477 NEZ F CvNl'N/I/O 1341 lr° i \ 0 Y r) N I Nu0 OR-1346 H3C"N NNIO 442 443 o N 0 y = 1342, NNJr-,, iN 492 493 1< S ° X NtO 1347 H3Cs, N) X 442 443 N'-O N\/0 1347 442 443 zozo HIC CH- H3C CH3 No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight S) I l N 0 Ny 0 1348 HC, N<O 460 461 1352 H3C N O 462 463 N : N1- 0 0 cl, N 0 1349 ç « 4 AZ9 HaCoN nNX l 9 ° | | [1353 1 t 527 t 28 4 N N 1353 527 528 0 o \ zon cl,) CHEZ N oH N oh o N , o 0 N'I I 1350 0 0 N 0 1354 H3C'NNJ"'. N 522 523 1y N 0 /I CH, /i I cHa 1355"3°N N 472 473 NEZ NCH3 y : 0 0 cl, 0/" <Yl / xi 0 OH y 458 459 non O /NQ f No MOLSTRUCTURE Weight S) No MOLSTRUCTURE Weight M CH 0 OH 0 N/O p'N 506 507 1362 CH3 X 474 475 N cl, chez N 0 N i O N I H' N CH, 1358 H3CNN<NX 472 473 1363 1 NtOXo 488 489 L J r ° "y-o i XCH3 H CNt CH, CH3 0OH CH3 lo / I 1359 H3C'N"-fO'*-N 472 473 1364 H3CI NIN 492 493 zon °0 Cl, \ y 0 0 N/0 N y/0 1360 H3CoN@ 490 491 H3r-I NI N ° 1365 II N° 557 558 1365 557 558 ° \ CH3 O-JO N 0 "- ou 1361 H3°N"N 458 459 II NO O/ N N 0 1366 N N 504 505 S O H3C CH3 No MOLSTRUCTURE Wetght M M No MOLSTRUCTtlRE A A-, Ns¢O H3CyCH3 NsrO H3C CH3 1367 N N . 54 455 1372 N' N 472 473 9-CH3 VNa ouzo c O- M H3'S, CH, , N 0 H3C CH- N 0 H3G'Chl3 1368 440 441 r HG/<NÇ4 1373 eN, N) X 440 441 N N N 0 H3CnC O 0 O Oli O N/0 H3C CH3 cl, 1369 NYO H3CYCH3 N"-y CH3 1369 g\N, Nn 488 489 oX 1 CH3 3 0 1374'IN 456 457 HC N 1 : C. CH, 1370 Nt X 454 455 N . N I I/CH 0 N 1375 I 470 471 XCH3 _ 1375 ORNoNk 470 471 /CHs.,,,,, . NO Zon H3C'v Uk Y 0 HC C 0 OH li 1371 N 454 455 Õ y : 0 Zon Foc CF No MOLSTRUCTURE . No MOLSTRUCTURE \ I \ I F F if N/ O H3C'CI-1 N//0 s r y 1376 H N NJ 474 475 1381 N N 590 591 HIC O \ I O /I/I F F fi 0 F N/O H3C CH3 N/0 r< ? _ H 0 1377 0l ° 539 540 0 c, , kif O''0 F OH N N N p Hc N N 556 557 0 0 o/ Sr° II I I O O 3^ l J 378 {q N)", zNx 606 607 cl, NYO N'J'-O kF F F N O rS _ 1384 c/<NX 574 575 Fun / . 0 N/0 I O 1379 N'N N 556 557 CH, N JE O I F F 0 CH3 kF CF Ny0 CH3 N/O F F 1385 H NN, 542 543 'i lu neo N O 1380 y 542 543 N CH N N 0 H3C CH3 H3C^CH ( ''0r' r 0 OH O OU N F F I , F No N.,, o/N O 1386 N 558 559 1391 NNN HCN/N 516 517 1 Wi 0 N I wl wl ON cl, 502 503 fY I"r HUG F nu F% N _ F Oh Ou// vil F Ny0 /I p N O Y 1393/eNoNs, N 550 551 1388 N N 576 577 HC vN N : 0 0 0 \/ F F N 0 N/0 I N 0 y'Y NN N 1394 NN 516 517 1389 If ° 641 642 ° o \ 0 0 0 0 N CH, 01-0 1395 NN 516 517 N I I N v'0 N 0 H rN0 HC) t !) : 1390 k 566 567 03C !) NO Ma. M+sH (M No MOLSTRUCTURE . No MOLSTRUCTURE Wetght N Ny0 N/O N O 1396 H NN 534 535 NEZ 1401 II ° 601 602 's ° \ 0 0 su 0 Oxo OH 'Y 1397 H NNN 502 503 I i H, 0 N y 0 HC4 , N NN 556 557 N C3 O OHM zizi / Y'N I \ I F 1398 N IN 0 518 519 N Zizi 1403 N'IN N 506 507 HC o T °' Y / r"-NIO N 0 OH N 0 H JS33 X \ I \ I N"'0 _ N 0 HC'CH, y 1400 HNN./N 536 537 N O j No MOLSTRUCTURE Weight M+H (M No MOLSTRUCTURE Weght M+HM s> s> /I F/ ! F \ N O N 0 1405 N-N-f'-**-N 540 541 1409 NIN N 492 493 NoSO 03 O9oN) ouzo 0 CL, F OH F 0 N 0 N 1406 N'IN N 506 507 1410 N IN 0 508 509 CH3 N1-o Ti O HC/ cl, F 3 N 0 Y 0 non \,/cl N N 0 0 oh 1407 NN N 506 507 1411 F I 522 523 HC J o I'''N'O N'y/O \ I N O _oH po /F 'IN /I I NO \ i fi 1408 H NNN 524 525 412 NN1IN 526 527 N 0 II I 524 ° 1' CH No MOLSTRUCTURE Wefght M+HM No MOLSTRUCTURE Weight M+H (M A A A No¢O W NtO y N 1418 HNNN 482 483 o.. io 1413 591 592 0'IN 0. O''0 CHZ N/O OH 1419 N'-r"-N 0 482 483 f')'Tr\ c 0 0 y I c 1f 0 Po N HC CH 1414 N, i o 532 533 C N O N O N'll'O 1 : NorO O ' 1 1415 NN"°\ 482 483 CH3 CH, _ No¢O 'IO CH3 Cl3 neo 1421 NI N 0 468 469 0 N No 1416 y 468 469 O A 0 0 NorO N0 1417/NNN° 516 517 - \ No MOLSTRUCTURE No MOLSTRUCTURE Weight OH 0 O N/O g HCNNN V N 0 T""OCOO 1422 J neo 484 485 X ; Nul ouzo OU 0 1 0 on i N HC/2 Nx _ CH2 1426 N N 518 519 \ No c CH N N i 1423 N 498 499 CH, O cl, 1427 N 468 469 HC Y 0 oh 0 my '--1 cH3 cl, CH3 N O N O 1424 N . 502 503 1428 J 454 455 it N H, C'CH, CFl H3C^CH3 I 1 K 1429 502 503 Hic'- Yo No MOLSTRUCTURE M. M+H (M No MOLSTRUCTURE S) Wefght S Weight \ CIi3 N-'O C CH N t N'NN 468 469 1435 ° 484 485 o I N o 0 \ ! ! CFI3 O , OH 10 con3/ At° cl, c 1 ? f r" 1 c 1431 N 468 469 N o H N N C a 3Cj 1436 HC/<NtX 488 489 N 0 HC'N 0 cl, /I CL, / N/O/\ (C. H3 CH3 1432 HC/<NX 486 487 Nt X 3 own N N zon 1437 553 554 cl, 0 ZU c o-0 0 ''0 CH3 1433 N"N 454 455 H2 oH 0 XßNCH, X X) J1 N O S 0/- 0 oh 0 cl3 CFL v'CH3 0 N1 i cru 1434 NNor° 470 471 tNtO X o N zizi 1439 N N Np 532 533 N 0 0 CF CF No MOLSTRUCTURE S) No MOLSTRUCTURE S) $ W ight O OHO/C _ 1440 CH3 o N \I N O I/O N 1440 518 519 1446 N1,,, ° 534 535 N N , xi < e A CH3 _ _ 1441 NIN N 566 567 H , N 0 1447 548 549 t N O N 0 / C FI3CO I/O OH zozo N,, rO CF 1442 Hc ( ( 532 533" fOT° ° 1448 1442 HC", N N y I 552 553 L. nez il \ CF CH3 _ t Ns¢O fJa A CIH3 1443 NIN N 532 533 N 0 Y 0 NIN N OH3C j H3 1449 WNa 617 618 0 = CF'IN \ I O O''0 1444 550 551 y-oh o OH o po S XI' . c I N o HC 0) 1450.'N 520 521 Nu0 neo 1445 N 0, 1445 N"N v 518 519 °cH N CH, y VNS _ cl CF CFt No MOLSTR UCTU RE Meight MvFH (M No MOLSTRUCTU RE Wel°ght M+H (h oCH3 N O N O 0- zu : Ny0 Ny0 HcN ./N 70 471 1456 N N 488 489 N o 0 H, 0 inc CL, S, CH, CL, MHz 1452 y 456 457 IN 1457 Nyo N 456 457 = f N v O H3C^CH3 p zu H, C CH, 0 OH CH, 0 N O . f NOC 1 N I 1453 N 504 505 0\/N1,,,,,. \3 1458 HCWJ 472 473 I Iw pCN3/ N/O J/I y 1 : T ° 1458 ° 472 473 _CH N 0 N 0 1459 0 N. CH3 C0 N N 0 T Y f ° 1459 ° f 486 487 N"0 J _ cl-f 0-- ( / 0 Ny0 1455 N N 470 471 CH, N NYO H, C yo -o-- 0 N O O c ' ? su i °-1 w o. c w o 1466 N N O/ N ; V. z. 9 Ct k. -NJ HC ! j C N-1140 H nez 07 \ OH N I/ o I 1467 HNNN 532 533 HC OH N0 1467 NINN 532 533 0 HC, _ 1462 582 583 0 c N O 0-i w ni I J4NX o 1468 ' ? iF 1468 N N0 H y N N N 532 533 0 Y Y ko HC S, CH, 0 cl, 0., CH,-------------------------------- AS (y 1469 5 519 HC 1464 y 518 519 N0 % NN N = HC H nô Cl, y = /O- N/ \ I 0 N 0 1470 N N0 534 535 1465 HNNN 566 567 N 0 y- O/ Mol M+H (M moi. M+H (M zero MHz /CL N N\/0 N 471 O N Ms 548 549 HCw X 518 519 0 r,"H "° N/\ 0 L. OH Y T Ills cul3 0 Ny0 Ny0 \ \ \ \ N O I N 0 1472 N/N N 552 553 1476 N N 504 505 f% Y CN 0"cl3 0---I hl3C^CH3 N 0 1 0 O o N 0/ y 1477 NN N 552 553 NIN N HC 1473 II ° 617 618 0 zizi N 0"cl, 0 Hz N O I/ OU 1478 N'N 518 519 O /, N 0 N O po 1474 N 568 569 N) 110 'CL CH Yo HIC N 0 C_ H, C No MOLSTRUCTURE No MOLSTRUCTURE /oCH3 I A, CH3 A, CH3 0 nu 0 my 1480 » <N nN 536 537 9 N 1485 XN 603 604 zon z CH3 !' 0)-0 n r 0'Cit H2 1481 vNoNnNJ 504 505 zozo H 0 po 6 538 539 N o I 0 oh Y'N OuN CH3 _ S iUi _ 1482 N'N y0 520 521 / N N \ HC f 147 H NNN 488 489 N H _ CH CH, YIN ha N 1483, o °N 534 535 N N' 0 0 ou 1488 N 474 475 p H N N Il nez 0 0 0H3C 11-ICH, N 0 'Y 1484 N--aN 538 539 - 0 0 j No MOLSTRUCTURE Weig. ht S) No MOLSTRUCTURE yMej9 ;" M+H (M Ny0 Ny0 N 0 \ f N O 1489 N'INN 522 523 1493 N NN 474 475 HC ; I 0 0 0 OH 0 Non 0 No w I O N,,, I I/ 1490 NY 488 489 1494 N."N0 490 491 \ -tao N 0 HC'--) N0 cl xi N O I j H 1491 NNN 488 489 1495 °,, N 504 505 ° i I N o zon Fc oui cit 0 OH 0 y 1492/<N'nN 506 507 NofO 9 H y 1496 N'9 508 509 0 N \CH3 O 0 No MOLSTRUCTURE _weight No MOLSTRUCTURE Weight Weight N 0 CFIa ZON 1497 o 573 574 HC, 454 455 1497 XN 573 574 XNA CF po L"1 ho W cl CH3 _ _ Oh Ny0 1503 N. IN 454 455 N N HC qu, N r v T Sr-o 1498 41N N 0 zozo Foc Chez /CH3 / i CH3 N No 1504 HCN'NN 472 473 N 1499 N 0 N N N y= 1 O (H3CH3 _ CH3 S, cru N 0 cl, N 0 1505 NINN 440 441 1500'^ 440 441 H % N N N N v 0 0 rN 0 OH /O NCH3 0 N C N-C YY' 1501 N'yo NY 488 489 1506 N'IN 0 456 457 z HC N No MOLSTRUCTURE Welghl M S (M No MOLSTRUCTURE"". M+H M wets ICH // 1512 464 465 N NIN'f""N") Co 1507 ° N 470 471 r'"" ° H3CN H3C^CH3 O / H vs 0 oh 0 Cl) 0 w 1513 1513 N-N-N 512 513 N O N O ou N 0 0 474 475 o o j i N N/O Ok 1514 H N'N N i | | W n CH 1514 NC » g9Hk XE > 478 4 9 N 00/y f 0 \ y z 0= m c CF N 1509 4 S 79 °"1 1 N Cl) N/0 1515 N 478 479 NO 0 ou I \ HaC /con3 0 O c 1510 H NN N 528 529 , N'\ O. i Nez . N 0 il ° 1 w I s. o N O fi 1511 rs11 9478 479 N O. o CH, CH, Y CH3 ; C * v X RUCTURE C No MOLSTRUCTURE Weight No MOLSTRUCTURE Welght S) Wel9nt S N y0 N 1517 NN 464 465 N'N-roo,-N I _ [1523 1 c&» K\N S I s7s 79 [ V gCH W ° 1521 Ii ° 563 564 'Ha N 0 O OH 0 zu CH OU OH N O N 1 N O 0., N. 'jt t o y N0 ro w i N NIT N.. '-' Zon 0 chez , cl N oh 3 CH N \ I N I/\ I v 1519 494 495 N nui 0 N O °, c^cH9 N , I N I wl /o 0 oH X. 1525 NNN 612 613 y : 0 Il xi N/O 1520 NNN /498 499 N w zon . XCH3 _ fol o 0 cH, No MOLSTRUCTURE Weghl M+HM No MOLSTRUCTURE Mo. M+H (M Welght ISV NX 3 aNn 3 _ Zon 1527 NIN I'N 578 579 NI N , N lf O HaCX 1533 o < 663 664 N f) 00 p I/ I 0''O ) j i) IN 596 597 ° S ° r" 0 SCH, N O po 1534 C : N, N N 607 608 Nllo ci 1529 564 565 0 ce c", I CI 0 oHO N N O O N ol CI N Cil 1530' 556 557 1530 NIN 01 oou 581 0 H ß O-CH3 \/CH3 cul CH I ce I N NN 1536 1531 10 N 594 595 Hc N 1531 N/\o N 0- 11 0 OH my 0 cri I N i 9. po fi l 1532 X 598 599 1537 591 592 0 H 0 0 No MOLSTRUCTURE Weight No MOLSTRUCTURE ' _ _ _ O8oOH N\¢O Wet9ht S 0 O O \ N O , I O N,, I I CI C/CI 1538 9, 556 557 1542 N 558 559 XCH d NIN 0 y-hic Ich CH, cul //j H N Cil I INN 1539 N 556 557 1543 ol ci 572 573 N C Nio 0 OH 1540 'OH CH3 -- _/I CI CI \/ I N \ I N CI CUL c Hcr HC N ! O 0 CL, cl ce /Np CI \ I N O CI HCN/NN 1541 H NNN 542 543 1545 II N° 642 643 N _ CH3 _ A X CH2 No MOLSTRUCTURE Wei 9h""S ""No MOLSTRUCTURE S) oh Cl, N 0N 0 0 r', y zizi N'O OI con3 nu') CH,, CFl3 CN NçO Õ 0 0. 0 0 Y 1547 NN f 456 457 1552 H N 474 475 N S ' 0 \/CN3 s CH3 a Ca I N 0 u 1548 Ny0 442 443 1553 IN f 442 443 H HC Hic II N II N ° 1 O oxo con N Nt f 0 N 0 N 1549 N N 490 49 1 1554 NN 0 457 458 n CON HC N _. _.. _ _--.-_- Cl3/ No o /CH bt, c H3 3 CH 550 v ;' : 5 | oANtS | 472 73 F oc pin N 0 0 OH 0 oh ° Y" No MOLSTRUCTURE Wefght M+H (M No MOLSTRUCTURE we Weight S) I I I N 0 0 N10 f'X/X, 7 A CH3 a ° XC Yo Yo O O CL, 0 I\ I CH3 \ HCN/N N 1562 H N cr 502 503 1557 0 ° 541 542 N o 0 0--l-0 CFl Ha \ OH ° 1563 0 1563 1563 NoNn i 502 503 N. I 558 M."oHdCH, 552 553 nI Y 1558 N HC c 1 'T'"Y 9 oc I 1564 H, 520 521 , N 1559 N 502 503 ana 0 0 0 CH," i S + 0 N0 1565 N"N H3 488 489 c HC 0 N CH, 0 No MOLSTRUCTURE Weight No MOLSTRUCTURE Weilht M+H (M 0 OH OH 0 CH 3 - 9 N 0 po 1566'IN0 1570"N 572 573 1566 504 505 N) 110 N N 0 y cul zon o cH 1571 H NNN 522 523 N N ZIZI N 1- 1567 0---N 518 519""r cr nif l _ CH, 0 OH 0 lu ° 1572 508 509 /H N N \ I//II NO H, N \ I Ha^'a 1568 HNNN cr 522 523 in 0 0 cl - N O C1 1573 HNNN 556 557 N /ici 0 o N O I i HCN N CH3 N 1569 587 588 N 0 cl 0 z y "N N9 522 523 1574 \ CL, No MOLSTRUCTURE Weight No MOLSTRUCTURE mol. M+H (M S) wl N/O I CI N I CI 1575 NN N 522 523 1580 NN N 542 543 Xi -0'Yo t t 0 0 il CL CRI 1576 H N'NN 540 541 y = 1581 607 608 o \ o CH, N ouzo i I Hz y cri OH f 1577 N N 508 509 Ho N o 0 po L N O 0 C ! J L i ci N cri cl zizi N 1583 NN N 556 557 HC 0 0 cl,, CH, ci I QN4Z X f 1 N 542 543 538 539 ,., y UL Y-° ! T 1 CCH, N O H , N T 0 CI 0 OH O No MOLSTRUCTURE No MOLSTRUCTURE Vyeight M S 0 oh cl ci \/ N CI N 0 I N 1/ N 1585 N'IN 558 559 O N ci w I ci ci , cl ich 1586 HNNN 556 557 i L M\ VF3C 1591 f {572 573 zon cl R LOH cri 0 oh my cl 0 cl 1587 Nf 4) _ _ WN i _ N y HCN/NN 576 577 O-L' v O ° cl, y cri /cl Cl 1588 X 575 578, < ,. 'N HCN/N N Ozon 0 o tCI 2 ci N zon I cl NO Hz 1589 N 542 543 H, H)-""'N 0 CL, I CI-I No MOLSTRUCTURE Weght M SM No MOLSTRUCTURE S) s ou /I CHa 0 \/ N N O I 1594 1599 Hc N'IN"a 502 503 N N 0 cl, 0ho II = CL, /ha CIi3 / I CH3 N N ta 1 1595 NIN N 502 503 y o 1600 HCNNN 520 521 O C II N O C 0 H" N 0 CH, 0 y : N O I I cl3 1596 488 489 N'N N N0 Hc ! L I N 0 1601 N N 488 489 N" HIC" 0 N H3C CH3 0 Clia O N /CH3 N O I 0 OH 0 1597 <N Ns s 536 537 0 nez 0 C 1602 sN Y 504 505 N HIC I CHU o kJ"o i J T 1598 N'502 503 N 1603 518 519 0 N'C/I"N'\0 I N CH T a No MOLSTRUCTURE Weioght No MOLSTRUCTURE S) Nof O 03 Nto X chai w, w I NO I/ N O y 1609 NIN Nf-C 1604 N 522 523 0 0 j cl, CL N0 1610 N'N y hui -0 1 'nez HCN N N O zon 1605 T i ° 587 588 k N ZIP \ \ XA At0 X / 1611 N N OH oh 0 p I Hs 0 cl 0 cl, 1606 HqN-N).,,,, _N 552 553 o wu O/ / IN-T : 520 521 . N ' ° 1 NO I S /CH3 1607 N 502 503 non CH, N O b, 1613 N N 488 489 /nu nez IN Nfl, 0 1608 y 488 489 NIN N H3C^CN3 No MOLSTRUCTURE Wefght M+HM No MOLSTRUCTURE Weight S) 0 OH OH 0 3 g OH o 'N 6,, 1618 580 581 1614 ' 504 505 [, f NYO hic N N N Ly")"o N zur N gCH 1619 s<NNz 530 531 HC N 1615 r,"N : L 0 CH, N N /O OH / N O N 516 517 /H N T N N 1 cl, Nu0 NYO 1616 Non'N 522 523 At0 X N v O = N O 1621 N Fr-N'-r-"-N 564 565 A'o-o II J-0 nif N Nue HC N 0 'Y 0 1622 N''IN 530 531 1"SW N ouzo cHZ \ y NsrO S CL, 1623 . NN 530 531 N 0 y : CHEZ cl, CH No MOLSTRUCTURE weig*ht No MOLSTRUCTURE NtO ß NtO X fi" . I\. Y 1624 H NN -N 548 549 1629 615 616 0 o CH, O Ny0 H2 1625 INN OH N-c4 516 517 N o \ N 0 \ NJ-,"N CN3 0 0 1630 N-, 552 553 O NX C O i N'-O Y 0' O N 1626 NNr 532 533 HIC" Nu0 0 0 g 1631-, c X sf3 502 503 % H I I I 0 , CH 0 H3 YIN 0 CF 1627 546 547 CF NEO OH N O 9 N o aH i o N o I 1632 488 489 HCN I N.. CH.' a 0 1628 550 551 ocH, tt'N 0 f f 1633 HC<N92 tCH3 536 537 N yin 0 No MOLSTRUCTURE Weght M+HM No MOLSTRUCTURE Weight M+HM N 0'c cl y N 1634'It"'Clt 502 503 1- N 1639 518 5 9 VNs/t t"N) o wl ou N 0 CF 0 OH 0 N O I/ 0 Hc 1640 NNN rcH, 522 523 lol 0 HC NO l-IG 'i H,, C') 0 CL3 1636 HCe < 520 521 1 ; t 1636 N XNSo 1641 9 X 587 588 o ouzo C"3 J nu0 oh 1637 HC N"N N.,,, CF6 488 489 0 N 0 jo N 1642 N 570 571 _ I I 0 OHO F a F N Y\ O N 1638 N'IN0 504 505 y Ho i I No MOLSTRUCTURE Weight S) No MOLSTRUCTURE F \ I F \ I F N 1 NtO 1643 NN N CF 520 521 1648,,,---N cH, 538 539 N II o II-o o c o Soc F N I I\ \I \ 1644 506 507 HC 1649 N C Nez F CIi3 \ O OH 0 cl3 N O/ 'Y 0 OH 1645 NNN CH 554 555 o HC N s,,, \ 1 1650 N, NtO 522 523 o w/, N N HC I\ F / N I 0 I/ 1646 N'NN cr 520 521 N CH 1651 O F N O ci CH3 O OH F X ? OL 537 . con 1647'IN 520 521 nu N F N 0 1652 9na 540 541 H, C'"C i J i o- 0 In addition, synthesis of the peptide mimetics of the library of the present invention may be accomplished using the General Scheme of [4,3, 0] Reverse-Turn Mimetic Library as follows : a Step 2-N I Step 3 Pol 0 Pol O RZ Pol O jyNHFmoc I I O H H N NHFmoc Step 4 'N\ N N 0 11 H I p 0 0 Re 0 R 0 RtNH_t Step 5 oN (\No R2 30 R6-N N O-. 0 5 P4 Synthesis of the peptide mimetics of the bicyclic template libraries of the present invention was accomplished using FlexChem Reactor Block which has 96 well plate by known techniques. In the above scheme'Pol'represents Bromoacetal resin (Advanced ChemTech) and detailed procedure is illustrated below.

Step 1 The bromoacetal resin (1. 6mmol/g) and a solution of R1 amine in DMSO (2M solution) were placed in 96 well Robbins block (FlexChem). The reaction mixture was shaken at 60°C using rotating oven [Robbins Scientific] for 12 hours. The resin was washed with DMF, MeOH, and then DCM Step 2 A solution of commercial available Fmoc-Amino Acids (4 equiv.), PyBob (4 equiv.), HOAt (4 equiv. ), and DIEA (12 equiv. ) in DMF was added to the resin. After the reaction mixture was shaken for 12 hours at room temperature, the resin was washed with DMF, MeOH, and then DCM.

Step 3 To the resin swollen by DMF before reaction was added 25% piperidine in DMF. After the reaction mixture was shaken for 30 min at room temperature. This deprotection step was repeated again and then washed with DMF, Methanol, then DCM. A solution of hydrazine carbamoyl chloride (4 equiv.), HOBt (4 equiv. ), and DIC (4 equiv. ) in DMF was added to the resin. After the reaction mixture was shaken for 12 hours at room temperature, the resin was washed with DMF, MeOH, and then DCM.

Step 4 To the resin swollen by DMF before reaction was added 25% piperidine in DMF. After the reaction mixture was shaken for 30 min at room temperature. This deprotection step was repeated again and then washed with DMF, Methanol, then DCM. To the resin swollen by DCM before reaction was added R1-isocynate (5 equiv.) in DCM. After the reaction mixture was shaken for 12 hours at room temperature the resin was washed with DMF, MeOH, then DCM.

Step 5 The resin was treated with formic acid (1.2 mL each well) for 18 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under reduced pressure using SpeedVac [SAVANT] to give the product as oil. These products were diluted with 50% water/acetonitrile and then lyophilized after freezing.

Table 3 shows a [4,3, 0] reverse turn mimetics library which can be prepared according to the present invention, of which representative preparation is given in Example 5.

TABLE 3 THE [4, 3, 0] REVERSE TURN MIMETICS LIBRARY 0 R, NH f N N, R2 R6-N ) r N fJ O ii O R4 4 No. R4 R6 R1 Mol. M+H Weight 610 lsoamyl 4-HO-phenyl Methyl Phenyl 466 467 611 Isoamyl 4-HO-phenyl Methyl 4-Me-phenyl 480 481 612 Isoamyl 4-HO-phenyl Methyl 3, 5-Me2-phenyl 494 495 613 Isoamyl 4-HO-phenyl Methyl 4-MeO-phenyi 496 497 614 Isoamyl 4-HO-phenyl Methyl 4-CF3-phenyl 534 535 615 Isoamyl 4-HO-phenyl Methyl Cyclohexyl 472 473 616 Isoamyl 4-HO-phenyl Methyl Benzyl 480 481 617 Isoamyl 4-HO-phenyl Methyl 494 495 iS 618 isoamyl 4-HO-phenyl Methyl 4-MeO-benzyl 510 511 619 Isoamyl 4-HO-phenyl Methyl Phenethyl 494 495 620 Isoamyl 4-HO-phenyl Methyl Pentyl 460 461 621 Isoamyl 4-HO-phenyl Methyl Hexyl 474 475 622 Benzyl 4-HO-phenyl Methyl Phenyl 486 487 623 Benzyl 4-HO-phenyl Methyl 4-Me-phenyl 500 501 624 Benzyl 4-HO-phenyl Methyl 3, 5-Me2-phenyl 514 515 625 Benzyl 4-HO-phenyl Methyl 4-MeO-phenyl 516 517 626 Benzyl 4-HO-phenyl Methyl 4-CF3-phenyl 554 555 627 Benzyl 4-HO-phenyl Methyl Cyclohexyl 492 493 628 Benzyl 4-HO-phenyl Methyl Benzyl 500 501 629 Benzyl 4-HO-phenyl Methyl 514 515 /S 630 Benzyl 4-HO-phenyl Methyl 4-MeO-benzyl 530 531 631 Benzyl 4-HO-phenyl Methyl Phenethyl 514 515 632 Benzyl 4-HO-phenyl Methyl Pentyl 480 481 633 Benzyl 4-HO-phenyl Methyl Hexyl 494 495 634 Naphth-1-ylmethyl 4-HO-phenyl Methyl Phenyl 536 537 635 Naphth-1-ylmethyl 4-HO-phenyl Methyl 4-Me-phenyl 550 551 636 Naphth-1-ylmethyl 4-HO-phenyi Methyl 3, 5-Me2-phenyl 564 565 637 Naphth-1-ylmethyl 4-HO-phenyl Methyl 4-MeO-phenyl 566 567 638 Naphth-1-ylmethyl 4-HO-phenyl Methyl 4-CF3-phenyi 604 605 639 Naphth-1-ylmethyl 4-HO-phenyl Methyl Cyclohexyl 542 543 640 Naphth-1-ylmethyl 4-HO-phenyl Methyl Benzyl 550 551 No R2 R4 R6 Ri Mol. M+H Weight 641 Naphth-1-ylmethyl 4-HO-phenyl Methyl 564 565 w 642 Naphth-1-ylmethyl 4-HO-phenyl Methyl 4-MeO-benzyl 580 581 643 Naphth-1-ylmethyl 4-HO-phenyl Methyl Phenethyl 564 565 644 Naphth-1-ylmethyl 4-HO-phenyl Methyl Pentyl 530 531 645 Naphth-1-ylmethyl 4-HO-phenyl Methyl Hexyl 544 545 646 Cyclohexylmethyl 4-HO-phenyl Methyl Phenyl 492 493 647 Cyclohexylmethyl 4-HO-phenyl Methyl 4-Me-phenyl 506 507 648 Cyclohexylmethyl 4-HO-phenyl Methyl 3, 5-Me2-phenyi 520 521 649 Cyclohexylmethyl 4-HO-phenyl Methyl 4-MeO-phenyl 522 523 650 Cyclohexylmethyl 4-HO-phenyl Methyl 4-CF3-phenyl 560 561 651 Cyclohexylmethyl 4-HO-phenyl Methyl Cyclohexyl 468 469 652 Cyclohexylmethyl 4-HO-phenyl Methyl Benzyl 506 507 653 Cyclohexylmethyl 4-HO-phenyl Methyl 520 521 w 654 Cyclohexylmethyl 4-HO-phenyl Methyl 4-MeO-benzyl 536 537 655 Cyclohexylmethyl 4-HO-phenyl Methyl Phenethyl 520 521 656 Cyclohexylmethyl 4-HO-phenyl Methyl Pentyl 486 487 657 Cyclohexylmethyl 4-HO-phenyl Methyl Hexyl 500 501 658 4-methylbenzyl 4-HO-phenyl Methyl Phenyl 500 501 659 4-methylbenzyl 4-HO-phenyl Methyl 4-Me-phenyl 514 515 660 4-methylbenzyl 4-HO-phenyl Methyl 3, 5-Me2-phenyl 528 529 661 4-methylbenzyl 4-HO-phenyl Methyl 4-MeO-phenyl 530 531 662 4-methylbenzyl 4-HO-phenyl Methyl 4-CF3-phenyl 568 569 663 4-methylbenzyl 4-HO-phenyl Methyl Cyclohexyl 506 507 664 4-methylbenzyl 4-HO-phenyl Methyl Benzyl 514 515 665 4-methylbenzyl 4-HO-phenyl Methyl 528 529 f, ~ 666 4-methylbenzyl 4-HO-phenyl Methyl 4-MeO-benzyl 544 545 667 4-methylbenzyl 4-HO-phenyl Methyl Phenethyl 528 529 668 4-methylbenzyl 4-HO-phenyl Methyl Pentyl 494 495 669 4-methylbenzyl 4-HO-phenyl Methyl Hexyl 508 509 670 Methoxypropyl 4-HO-phenyl Methyl Phenyl 468 469 671 Methoxypropyl 4-HO-phenyl Methyl 4-Me-phenyl 482 483 672 Methoxypropyl 4-HO-phenyl Methyl 3, 5-Me2-phenyl 496 497 673 Methoxypropyl 4-HO-phenyl Methyl 4-MeO-phenyl 498 499 674 Methoxypropyl 4-HO-phenyl Methyl 4-CF3-phenyl 536 537 675 Methoxypropyl 4-HO-phenyl Methyl Cyclohexyl 474 475 676 Methoxypropyl 4-HO-phenyl Methyl Benzyl 482 483 677 Methoxypropyl 4-HO-phenyl Methyl 496 497 fas 678 Methoxypropyl 4-HO-phenyl Methyl 4-MeO-benzyl 512 513 679 Methoxypropyl 4-HO-phenyl Methyl Phenethyl 496 497 680 Methoxypropyl 4-HO-phenyl Methyl Pentyl 462 463 No R2 R4 R6 Ri Mol. M+H Weight 681 Methoxypropyl 4-HO-phenyl Methyl Hexyl 476 477 682 Phenethyl 4-HO=phenyl Methyl Phenyl 500 501 683 Phenethyl 4-HO-phenyl Methyl 4-Me-phenyl 514 515 684 Phenethyl 4-HO-phenyl Methyl 3, 5-Me2-phenyl 528 529 685 Phenethyl 4-HO-phenyl Methyl 4-MeO-phenyl 530 531 686 Phenethyl 4-HO-phenyl Methyl 4-CF3-phenyl 568 569 687 Phenethyl 4-HO-phenyl Methyl Cyclohexyl 506 507 688 Phenethyl 4-HO-phenyl Methyl Benzyl 514 515 689 Phenethyl 4-HO-phenyl Methyl 528 529 iC 690 Phenethyl 4-HO-phenyl Methyl 4-MeO-benzyl 544 545 691 Phenethyl 4-HO-phenyl Methyl Phenethyl 528 529 692 Phenethyl 4-HO-phenyl Methyl Pentyl 494 495 693 Phenethyl 4-HO-phenyl Methyl Hexyl 508 509 694 2, 2-bisphenylethyl 4-HO-phenyl Methyl Phenyl 576 577 695 2, 2-bisphenylethyl 4-HO-phenyl Methyl 4-Me-phenyl 590 591 696 2, 2-bisphenylethyl 4-HO-phenyl Methyl 3, 5-Me2-phenyl 604 605 697 2, 2-bisphenylethyl 4-HO-phenyl Methyl 4-MeO-phenyl 606 607 698 2, 2-bisphenylethyl 4-HO-phenyl Methyl 4-CF3-phenyl 644 645 699 2, 2-bisphenylethyl 4-HO-phenyl Methyl Cyclohexyl 582 583 700 2, 2-bisphenylethyl 4-HO-phenyl Methyl Benzyl 586 587 701 2, 2-bisphenylethyl 4-HO-phenyl Methyl 604 605 fC 702 2, 2-bisphenylethyl 4-HO-phenyl Methyl 4-MeO-benzyl 620 621 703 2, 2-bisphenylethyl 4-HO-phenyl Methyl Phenethyl 604 605 704 2, 2-bisphenylethyl 4-HO-phenyl Methyl Pentyl 570 571 705 2, 2-bisphenylethyl 4-HO-phenyl Methyl Hexyl 584 585 706 Naphth-1-ylmethyl Benzyl Methyl Phenyl 520 521 707 Naphth-1-ylmethyl Benzyl Methyl 4-Me-phenyl 534 535 708 Naphth-1-ylmethyl Benzyl Methyl 3, 5-Me2-phenyl 548 549 709 Naphth-1-ylmethyl Benzyl Methyl 4-MeO-phenyl 550 551 710 Naphth-1-ylmethyl Benzyl Methyl 4-CF3-phenyl 588 589 711 Naphth-1-ylmethyl Benzyl Methyl Cyclohexyl 526 527 712 Naphth-1-ylmethyl Benzyl Methyl Benzyl 534 535 713 Naphth-1-ylmethyl Benzyl Methyl 548 549 w 714 Naphth-1-ylmethyl Benzyl Methyl 4-MeO-benzyl 564 565 715 Naphth-1-ylmethyl Benzyl Methyl Phenethyl 548 549 71 E Naphth-1-ylmethyl Benzyl Methyl Pentyl 5i4 515 717 Naphth-1-ylmethyl Benzyl Methyl Hexyl 528 529 718 Naphth-1-ylmethyl Methyl Phenyl 498 499 spirit O 719 Naphth-1-ylmethyl Methyl 4-Me-phenyl 512 513 spiro O 720 Naphth-1-ylmethyl Methyl 3, 5-Me2-phenyl 526 527 spiro ' 721 Naphth-1-ylmethyl Methyl 4-MeO-phenyl 528 529 spiro O 722 Naphth-1-ylmethyl Methyl 4-CF3-phenyl 566 567 spiro - 723 Naphth-1-ylmethyl Methyl Cyclohexyl 504 505 spirit O 724 Naphth-1-ylmethyl Methyl Benzyl 512 513 spiro \ 725 Naphth-1-ylmethyl Methyl i I 526 527 i,, fS -spiro l 726 Naphth-1-ylmethyl Methyl 4-MeO-benzyl 542 543 spiro 727 Naphth-1-ylmethyl Methyl Phenethyl 526 527 spiro 728 Naphth-1-ylmethyl Methyl Pentyl 492 493 spire 0 1 sPa o 729 Naphth-1-ylmethyl Methyl Hexyl 506 507 spiro O 730 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl Phenyl 570 571 731 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl 4-Me-phenyl 584 585 732 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl 3, 5-Me2-phenyl 598 599 733 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl 4-MeO-phenyl 600 601 734 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl 4-CF3-phenyl 638 639 735 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl Cyclohexyl 576 577 736 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl Benzyl 584 585 737 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl 598 599 fus 738 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl 4-MeO-benzyl 614 615 739 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl Phenethyl 598 599 740 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl Pentyl 564 565 741 Naphth-1-ylmethyl Naphth-1-ylmethyl Methyl Hexyl 578 579 742 Naphth-1-ylmethyl Cyclohexylmethyl Methyl Phenyl 526 527 743 Naphth-1-ylmethyl Cyclohexylmethyl Methyl 4-Me-phenyl 540 541 744 Naphth-1-ylmethyl Cyclohexylmethyl Methyl 3, 5-Me2-phenyl 554 555 745 Naphth-1-ylmethyl Cyclohexylmethyl Methyl 4-MeO-phenyl 556 557 746 Naphth-1-ylmethyl Cyclohexylmethyl Methyl 4-CF3-phenyl 594 595 747 Naphth-1-ylmethyl Cyclohexylmethyl Methyl Cyclohexyl 532 533 748 Naphth-1-ylmethyl Cyclohexylmethyl Methyl Benzyl 540 541 749 Naphth-1-ylmethyl Cyclohexylmethyl Methyl 554 555 iH 750 Naphth-1-ylmethyl Cyclohexylmethyl Methyl 4-MeO-benzyl 570 571 751 Naphth-1-ylmethyl Cyclohexylmethyl Methyl Phenethyl 554 555 752 Naphth-1-ylmethyl Cyclohexylmethyl Methyl Pentyl 520 521 753 Naphth-1-ylmethyl Cyclohexylmethyl Methyl Hexyl 534 535 754 Naphth-1-ylmethyl 4-chlorobenzyl Methyl Phenyl 554 555 755 Naphth-1-ylmethyl 4-chlorobenzyl Methyl 4-Me-phenyl 568 569 756 Naphth-1-ylmethyl 4-chlorobenzyl Methyl 3, 5-Me2-phenyl 582 583 757 Naphth-1-ylmethyl 4-chlorobenzyl Methyl 4-MeO-phenyl 584 585 758 Naphth-1-ylmethyl 4-chlorobenzyl Methyl 4-CF3-phenyl 622 623 759 Naphth-1-ylmethyl 4-chlorobenzyl Methyl Cyclohexyl 560 561 760 Naphth-1-ylmethyl 4-chlorobenzyl Methyl Benzyl 568 569 761 Naphth-1-ylmethyl 4-chlorobenzyl Methyl n 582 583 i 762 Naphth-1-ylmethyl 4-chlorobenzyl Methyl 4-MeO-benzyl 598 599 763 Naphth-1-ylmethyl 4-chlorobenzyl Methyl Phenethyl 582 583 764 Naphth-1-ylmethyl 4-chlorobenzyl Methyl Pentyl 548 549 765 Naphth-1-ylmethyl 4-chlorobenzyl Methyl Hexyl 562 563 766 Naphth-1-ylmethyl Methyl Methyl Phenyl 444 445 767 Naphth-1-ylmethyl Methyl Methyl 4-Me-phenyl 458 459 768 Naphth-1-ylmethyl Methyl Methyl 3, 5-Me2-phenyl 472 473 769 Naphth-1-ylmethyl Methyl Methyl 4-MeO-phenyl 474 475 770 Naphth-1-ylmethyl Methyl Methyl 4-CF3-phenyl 512 513 771 Naphth-1-ylmethyl Methyl Methyl Cyclohexyl 450 451 772 Naphth-1-ylmethyl Methyl Methyl Benzyl 458 459 773 Naphth-1-ylmethyl Methyl Methyl 472 473 i 774 Naphth-1-ylmethyl Methyl Methyl 4-MeO-benzyl 488 489 775 Naphth-1-ylmethyl Methyl Methyl Phenethyl 472 473 776 Naphth-1-ylmethyl Methyl Methyl Pentyl 438 439 777 Naphth-1-ylmethyl Methyl Methyl Hexyl 452 453 778 Naphth-1-ylmethyl Isobutyl Methyl Phenyl 486 487 779 Naphth-1-ylmethyl Isobutyl Methyl 4-Me-phenyl 500 501 780 Naphth-1-ylmethyl Isobutyl Methyl 3, 5-Me2-phenyl 514 515 781 Naphth-1-ylmethyl Isobutyl Methyl 4-MeO-phenyl 516 517 782 Naphth-1-ylmethyl Isobutyl Methyl 4-CF3-phenyl 554 555 783 Naphth-1-ylmethyl Isobutyl Methyl Cyclohexyl 492 493 784 Naphth-1-ylmethyl Isobutyl Methyl Benzyl 500 501 785 Naphth-1-ylmethyl Isobutyl Methyl 515 fas 786 Naphth-1-ylmethyl Isobutyl Methyl 4-MeO-benzyl 530 531 787 Naphth-1-ylmethyl Isobutyl Methyl Phenethyl 514 515 788 Naphth-1-ylmethyl Isobutyl Methyl Pentyl 480 481 789 Naphth-1-ylmethyl Isobutyl Methyl Hexyl 494 495 790 Naphth-1-ylmethyl Methylthioethyl Methyl Phenyl 504 505 791 Naphth-1-ylmethyl Methylthioethyl Methyl 4-Me-phenyl 518 519 792 Naphth-1-ylmethyl Methylthioethyl Methyl 3, 5-Me2-phenyl 532 533 793 Naphth-1-ylmethyl Methylthioethyl Methyl 4-MeO-phenyl 534 535 794 Naphth-1-ylmethyl Methylthioethyl Methyl 4-CF3-phenyl 572 573 795 Naphth-1-ylmethyl Methylthioethyl Methyl Cyclohexyl 510 511 796 Naphth-1-ylmethyl Methylthioethyl Methyl Benzyl 518 519 797 Naphth-1-ylmethyl Methylthioethyl Methyl 532 53 fAS 798 Naphth-1-ylmethyl Methylthioethyl Methyl 4-MeO-benzyl 548 549 799 Naphth-1-ylmethyl Methylthioethyl Methyl Phenethyl 532 533 800 Naphth-1-ylmethyl Methylthioethyl Methyl Pentyl 498 499 801 Naphth-1-ylmethyl Methylthioethyl Methyl Hexyl 512 513

In a further aspect of this invention, the present invention provides methods for screening the libraries for bioactivity and isolating bioactive library members.

In yet another aspect, the present invention provides a method for carrying out a binding assay. The method includes providing a composition that includes a first co-activator, an interacting protein, and a test compound.

The amino acid structure of the first co-activator includes a binding motif of LXXLL, LXXLI or FxxFF wherein X is any amino acid. The method further includes detecting an alteration in binding between the first co-activator and the interacting protein due to the presence of the compound, and then characterizing the test compound in terms of its effect on the binding.

The assay may be carried out by any means that can measure the effect of a test compound on the binding between two proteins. Many such assays are known in the art and can be utilized in the method of the present invention, including the so-called Two-Hybrid and Split-Hybrid systems.

The Two-Hybrid system, and various means to carry out an assay using this system, are described in, e. g. , U. S. Patent 6,410, 245. The Split-

Hybrid system has been described by, e. g., Hsiu-Ming Shiu et al. Proc. Natl.

Acad. Sci. USA, 93: 13896-13901, November 1996; and John D. Crispino, et al.

Molecular Cell, 3: 1-20, February 1999. In the Split-Hybrid system, a fusion protein is utilized where protein X is fused to the lexA DNA binding domains (pLexA) and protein Y is fused to the transcription activator VP16 (pSHM. 1- LacZ). Interaction between lexA-X and VP16-Y leads to the expression of the Tetracycline repressor protein (TetR). TetR prevents transcription of the HIS3 reporter gene, making the cells unable to grow on media lacking histidine.

Disruption of protein-protein interaction will restore the ability of the cells to grow on such media by shutting down expression of the tetracycline repressor.

Accordingly, compounds of the present invention may be added to the growing cells, and if the addition of the compound restores the ability of the cells to grow on the media, the compound may be seen as an effective disruptor of the protein-protein interaction.

The yeast strains required to make the Split-Hybrid system work can be employed with two hybrid LexA/VP16 constructs such as those described by Stanley M. Hollenberg, et al. Molecular and Cellular Biology 15 (7): 3813-3822, July 1995. A useful modification of the Split-Hybrid system was utilized by Takemaru, K. 1. and Moon, R. T. J. of Cell Biol. 149: 249-254, 2000.

Other assay formats are also suitable. For example, reporter gene assays for AP-1, ELISA, for example, blocking the production of IL-2 by a T-cell line after stimulation with CD3 and CD28 to look for inhibitors of IL-2 transcription. Direct binding assays (between coactivators and their partners) can be performed by surface plasmon resonance spectroscopy (Biacore, Sweden, manufactures suitable instruments) or ELISA.

Exemplary transcriptional regulators include, without limitation, VP16, VP64, p300, CBP, PCAF, SRC1 PvALF, AtHD2A and ERF-2. See, for example, Robyr et al. (2000) MoL Endocrinol. 14 : 329-347 ; Collingwood et al.

(1999) J. Mol. Endocrinol. 23: 255-275; Leo et al. (2000) Gene 245 : 1-11;

Manteuffel-Cymborowska (1999) Acta Biochim. Pol. 46: 77-89; McKenna et al.

(1999) J. Steroid Biochem. Mol. Biol. 69: 3-12; Malik et al. (2000) Trends Biochem. Sci. 25: 277-283; and Lemon et al. (1999) Curr. Opin. Genet. Dev.

9: 499-504. Other exemplary transcription factors include, without limitation, OsGAI, HALF-1, C1, AP1, ARF-5, -6,-7, and-8, CPRF1, CPRF4, MYC-RP/GP, and TRAB1. See, for example, Ogawa et al. (2000) Gene 245: 21-29; Okanami et al. (1996) Genes Cells 1: 87-99; Goff et al. (1991) Genes Dev.

5: 298-309; Cho et al. (1999) Plant Mol. Biol. 40: 419-429; Ulmason et al. (1999) Proc. Natl. Acad. Sci. USA 96: 5844-5849; Sprenger-Haussels et al. (2000) Plant J. 22: 1-8; Gong et al. (1999) Plant Mol. Biol. 41: 33-44; and Hobo et al.

(1999) Proc. Natl. Acad. Sci. USA 96: 15, 348-15, 353.

In a preferred embodiment, the transcriptional coactivator is a human transcriptional coactivator. In another preferred embodiment, the transcriptional coactivator is a member of the p300/CBP family of co-activators which have histone acetyltransferase activity. p300 is described for example by Eckner et al, 1994 and CBP by Bannister and Kouzarides, 1996. For the purposes of the present invention, reference to p300/CBP refers to human allelic and synthetic variants of p300, and to other mammalian variants and allelic and synthetic variants thereof, as well as fragments of said human and mammalian forms of p300. In one aspect of the assay, the interacting protein is a transcription factor or a second co-activator.

In one aspect of the assay, the interacting protein is any one of RIP140 ; SRC-1 (NCoA-1); TIF2 (GRIP-1 ; SRC-2); p (CIP ; RAC3; ACTR; AIB-1 ; TRAM-1; SRC-3); CBP (p300); TRAPs (DRIPs) ; PGC-1; CARM-1; PRIP (ASC- 2; AIB3 ; RAP250; NRC); GT-198; and SHARP (CoAA ; p68; p72). In another aspect of the assay, the interacting protein is any one of TAL 1; p73; MDm2; TBP; HIF-1 ; Ets-1; RXR; p65; AP-1; Pit-1; HNF-4 ; Stat2 ; HPV E2; BRCA1 ; p45 (NF-E2); c-Jun; c-myb; Tax; Sap 1; YY1 ; SREBP; ATF-1; ATF-4 ; Cubitus; Interruptus ; Gli3 ; MRF; AFT-2; JMY; dMad ; PyLT : HPV E6; CITTA ; Tat; SF-1; E2F; junB ; RNA helicase A; C/EBP ß ; GATA-1; Neuro D; Microphthalimia ; E1A ;

TFIIB ; p53; P/CAF; Twist; Myo D; pp90 RSK; c-Fos; and SV40 Large T. In another aspect of the assay, the interacting protein is any one of ERAP140; RIP140 ; RIP160 ; Trip1 ; SWI1 (SNF); ARA70; RAP46; TIF1 ; TIF2 ; GRIP1 ; and TRAP. In another aspect of the invention, the interacting protein is any one of VP16; VP64; p300 ; CBP; PCAF; SRC1 PvALF ; AtHD2A; ERF-2; OsGAI ; HALF- 1; C1 ; AP-1; ARF-5; ARF-6; ARF-7; ARF-8; CPRF1 ; CPRF4; MYC-RP/GP; and TRAB1. In another aspect of the invention, the first co-activator is CBP or p300.

The test compound is selected from compounds as described herein. For example, compounds having the formula (1), (II), (111), (IV), (Vl) and (Vla). Typically, a test compound will be evaluated at several different concentrations, where these concentrations will be selected, in part, based on the conditions of the assay, e. g. , the concentrations of the first co-activator and the interacting protein. Concentrations in the range of about 0.1 to 10 uM are typical. In one aspect, the assay evaluates the relative efficacy of two compounds to affect the binding interaction between two proteins, where at least one of those two compounds is a compound of the present invention.

The more effective compound can than serve as a reference compound in a study of the relationship between compound structure and compound activity.

The libraries of the present invention were screened for bioactivity by various techniques and methods. In general, the screening assay may be performed by (1) contacting the mimetics of a library with a biological target of interest, such as a receptor, to allow binding between the mimetics of the library and the target to occur, and (2) detecting the binding event by an appropriate assay, such as the calorimetric assay disclosed by Lam et al. (Nature 354: 82- 84,1991) or Griminski et al. (Biotechnology 12: 1008-1011, 1994) (both of which are incorporated herein by reference). In a preferred embodiment, the library members are in solution and the target is immobilized on a solid phase.

Alternatively, the library may be immobilized on a solid phase and may be probed by contacting it with the target in solution.

Table 4 below shows compounds for bioactivity test selected from the library of the present invention and IC50 values thereof, which are measured by the Reporter gene assay as described in Example 6.

TABLE 4 IC50 (pM) OF SELECTED LIBRARY COMPOUNDS No STRUCTURE M. W. IC50 (M) CF Y FEZ 0Y0 F o 580. 7 12. 8 o F FI I F F N O 2 y 5 79. 6 12. 6 NIN ZON ici O H3CuNa 0 CF 3 N 0 OH 632. 5 13. 9 Nu) a I CI'Y I-ICN v O CI FF F I non NC, NINN O OH No STRUCTURE M. W. X M/) V NU ru OY"N11 -OH 5 H3CuNtX 564. 6 6. 8 0 rol ZON \ 6 H3Cu<1 564. 6 6. 1 0 0 \ X- T N11 'N \ 7 H3CN44 JD 564. 6 2. 2 O OH zou =o N 531. 6 F NYO ou N''0 2 0 0. 9 F 531. 6 6. 7 NO s No STRUCTURE M. W. ^ OH =M. / ? O ou \ N NNwcH, 531. 6 4. 0 Nô 0H. I., oH a9 F =o zt ans su 531. 6 4. 6 neo =o N F F 0.. 7all =_o 13 13 N"'CH, 549. 6 6. 4 1 2 NS/tN'NCH 54 9. 6 9. O 13 N9\N'NCH 549. 6 6. 4 | 14 F<CC j 549 6 , 10 I =o 14 neo dz NYO No STRUCTURE M. W. IC50 (M) OU =o 0 N 15 FA >, b CH3 581. 6 4. 2 NO Oh N' 0 N .-. N F \ I 0/ w 16 F tCH3 567. 6 3. 8 NO OH ^'OH ^° / Z7 , 548. 0 14. 3 cl I\ Ruz I/ =o r4 N 18 a'NcH, 548. 0 3. 3 C So I N'O cri =o 0 19 19 N''Cit 582. 5 11. 5 NO NoSTRUCTUREM. W. tCsoM) : O Po 20 u ; 527. 6 5. 1 CN3 NYO . 0 : O 0 N . N CN3 527. 6 5. 0 OH 0 e Oh : 0 = O 22 543. 6 10. 4 NYO au =o p N EFC, o ' 573. 6 10. 7 CH, NO ON i 0 N 24"° N 563. 7 5. 0 o bzw OH No STRUCTURE M. W. IC5o (, M) I Oq _ y. 25 5 H3CN 5 81. 6 3. 0 ZIZI yin 0 0 Zou I N\ jo oc 26 H3CNt X 543. 6 7. 1 0"ah OH i OH I 0 o, CH3 27 H3CNt J 54 3. 6 5. 2 OH 0 riz OH N 28 b to/% 548. 0 7. 5 0 ci cm bzw v OH 1 : y0 29 "N-YY"582. 5 3. 8 « ci zou zizi OH No STRUCTURE M. W. tCepM) I F 0 F-F 0''F 30 NA, 597. 6 7. 5 0 OH v'oh sl F y0 S _F 31 H3CoN<-613. 7 11. 9 ZOU 0 OH F N O F F F 32 Nus 561. 6 4. 1 0 y : 0= I NtO N10 33 % N 4 564. 6 13. 0 I zou oh /I ""aOH Y 0 y N 34 ftc-I N'IN)""NN 565. 6 4. 4 ° Sci O \ v qu 0 {UCTURE M. W. ICso (pM) 1 N, fus N/S 35 N 579. 7 11. 4 o v n (' OH I 5 S ; 549. 6 12. 5 N V cl ° r oh il H3C-1 h 37 7 D F 545. 6 2. 3 po OH I/ Cf \ N 0 Na 38 H3CuNX 3/sCH3 556. 7 7. 1 - o 'OU zu N i0 | n t 564. 6 9. 7 oDj SOH No STRUCTURE M. W. tCsoM) Ny 0 N O 'NN 553. 6 7. 0 Nez OH O \ ou ON T'CH3 41 H'°N'N j 541. 6 13. 6 - 0 CH3 OH OH W iN I 42 a 1 574. 7 18. 2 O ou 0 No¢O H3CvNCH3 43 NCIN'.'-fll"-N 556. 7 5. 2 OU I v'ON s No¢O F F _f 44 H, C, NN, N 599. 6 1. 3 N F y = 0 = No STRUCTURE M. W. tCsoM) xi Ns¢O H3CuNCH3 N 45 CNNN \ 591. 1 2. 2 Nez zou il aOH v OH if 46 H3CuNX'CH3 570. 7 4. 4 0 ", ah ni CF N O'CN3 47 H3C, NIN'-'CN 584. 17 3. 5 0 II N0 OU N I O wNi 48 N-Y\"570. 7 10. 9 O II N (sl I 0 v oh N0 No ; O sN' 49-N'N- 592. 6 1. 4 s OF OH po \ N I O Ni 50 sN e 574 6 1. 3 O I F o = OH No STRUCTURE M. W. IC50 (RM) N I O N 51 uN9 584. 7 4. 8 O po v'oh Ny 0 N/O 52 N 621. 69 25 o o OH 0 Chl. 1 Ny 0 N"CH, I 53 H3CuN 584. 72 9. 0 1. 5 /nez zou OH CH3CCH3"m J A CHa N\/0/ CH3 54 H3CoNCo 619. 16 23. 6 5. 6 N °'y 0 I OH No STRUCTURE M. W. JCsoM) CHw cl, \ I \ NwCHa N\/0/ 55 H RN, N) to 584. 72 7. 2 + 1. 4 N-1o OH 0 'Y"r' Oh N 0 f N 56 H3CuNgN_CH3 567. 65 9. 3 _ 1. 6 N OH tri "-ah v'OH omai X No¢O Y 57 H'NNrnt I \ NCH3 582. 70 9. 4 _ 1. 5 zozo 0 OH oman N i0 CN CN 58 Nv-cH, 588. 68 49. 1 8. 1 ZON Zou 0 'OH NoSTRUCTUREM. W. tCspM) Moo N i O Y CH3 ICH3 59 H3CoN sCH3 588. 68 5. 3 1. 3 Zou v'oh 0 NYO Nyo CH, CH 60 NYYcH, 638. 69 6. 9 1. 7 ° ^ 0 F F F OU OU m Hs 61 H3CNlitNCH3 570. 69 25. 8 nez Y : 'OU OH N. CH I-IC,/CHa _ 3 62 H3C"CH, 616. 73 9. 7 1. 7 N nO Zip 'oh \ 63 N-NW" 582. 70 4. 1 0. 5 'oh , aOH No STRUCTURE M. W. IC50 (M) MHz CH3 J H3 N/O . li CH3\N 64 H3C, N N 11 616. 73 25. 3 6. 6 /N 0 F T 0 OH OH \ I CNwCHs N, CH 3 N o 65 H 3CI N'IN N CH3 616. 73 19 7. 1 /nez zozo ou OH Y CN3 i H3 I 66 H3C, NIIN Ny CH3 598. 74 11. 8 N 0 0 Zip 'oh in f'Chl i lia 67 H3CI N. IN)-*Io-N N CH 598. 74 6. 8 N y OH 0 , \ I/ oh No STRUCTURE M. W. IC50 (pM) Ny 0 68 r, w 590. 68 4. 3 + 0. 8 /nez il 0 Chihzl NYO ' H CN3 F 69"NNY 563. 60 1. 4 0. 7 /N OF 0 F y : ou , aOH 10 C"" 'Y-N 70 H3CNt N 553. 62 8. 8 1. 9 0 0 OH N y 0 OH Y 71 ç, NS 596. 73 6. 5 0. 7 XI 'OU No STRUCTURE M. W. tCsoM) Cher l \ I HaN N\/0 72 H 3CINNN 658. 76 1. 6 0. 1 11- 0 0 Oh on han Ny N-N 73 H3CuN 658. 76 3. 6 po 0 oh Oh cam Ny ° su OH N-N : 74"3NN I w 688. 74 2. 1 0. 2 N/ OU Zou Chlral No N-N 75 f t C 566. 64 50. 5 18. 4 N, r, 0 0 ou No STRUCTURE M. W. Y° N0 'Y N=N 76 NYY 568. 64 10. 7 2. 5 0 OH z 'oh cnm fi 77 NY-s 570. 67 7. 2 2. 5 '% N OH Nez OH /oh I N O H, C, N N 78 N'"1-*"4N-cH. 570. 69 4. 3 0. 9 /N ", ah Ici O 'OH cnm 79 NN 632. 76 16. 5 4. 8 o 0 0 Oh , , - N/O . Y C I . ì tAi u CH i 605. 14 7. 9 + 2. 0 | N : 0 ci y 'oh No STRUCTU RE M aW. lC50 (M) A Chlml kjk N/0/0 'Y F 81 81 m ì t m. 61 66. 1 6. 8 OF OH OH hW Ch, eì . f F i 82 ; D t l F 579. 60 68. 1 8. 9 Y OH ",-aOH N 0 I\ oh 1 N 0 a 83 H3CoN tCIsCH3 605. 14 46. 4 3. 7 0 cri 'OH \OH v n NYN/V 0 740. 79 46. 7 6. 7 0 I CHEZ CHEZ C UCTURE M. W. ICso (MM) ni 0 'Y° f 85 HC, NN , N 549. 67 15. 6 2. 2 0 OH on Fizz N 0 roll 86 i 'OU cnW fl r Chl. 1 NYO 87 H3CN-624. 74 8. 1 0. 8 'nez OH cl. l Oh 88 N i N t bNH 658. 76 2. 2 i 0. 2 II N0 I OH v'OH No STRUCTURE M. W. lC5o (UM) ont /N /I I \ 553. 62 13. 9 0. 9 /nez 1 O I-r 0 oh OH N CM v 90 wn NH2 647. 78 3. 9 f Chlnl -, AH o 1 : Oh NH2 hm l v N 0 N-N 91 H3c, NINN 658. 76 2. 9 0. 2 I 0 RIZ OH Han /\ AY° N-N 92 H3CuNv 658. 76 3. 8 1. 2 o N OH No STRUCTURE M. W. ! IC50 (RM) 'y° 93 H3CI N. N N\ 591. 67 6. 8 1. 3 N Yin 0 0 ", aoH cnm, 0 xi H3C, y 'Y N-N 94 H3CuNv 666. 78 7. 6 + 0. 6 N, r OH zu ou Chl, l N/O 564. 64 13. 3 1. 4 / 0 ""ah nui can ,, H N/0 y 95 F X 59 67 8. +0. 9 N 0 0 ""aOH OH No STRUCTURE M. W. tCsoM) C"" N 0 (0) T N 97 H3NNN \ 598. 70 12. 6 1. 2 A ChlMl 0 OH riz oh zon 98 H3C, N 666. 78 14. 4 2. 2 Yc 0 V'OH N 0 99 H'C N'Y N \ N 0 'OH D 0" 701. 78 2. 4 0. 3 o N ICH3 I-r Q ii N/O XOH OH 0 No p cnm N/0 101 H3C, 666. 78 3. 9 06 0 , aOH No STRUCTURE M. W. tCgoM) Cl Ny 0 102 Y''TT S11. 58 62. 0 17. 0 At° F 0 Oui OH Chlral N 0 y F ZOU 103 H3NNN I 535. 59 14. 5 1. 7 0 = "-aOH 0 ou NHZ °°m N 0 N-N 104 H, C, NIIN N 658. 76 4. 6 0. 4 0 oh C"" HC ou m / I HC y N-N 105 H3CuNt 591. 67 16. 6 2. 7 0 0 OH O OH No STRUCTURE M. W. IC50 (ZM) No¢O N/ ,, cl / N O 'Y N 106 HC, N. NN 591. 67 2. 6 0. 2 'nez 0 OU 'on N Y/0 a i 107 X, P 724. 82 2. 7 0. 3 OH | o cl "'OH Chlnl Nof O H3CuNCH3 N. 0 H, CCH, 108 N'TaN 616. 67 1. 6 0. 1 yin | S f C"" Chtra) Nsr FCIN H3CN'CH3 OH omm, N 109 N N 616. 67 2. 1 N _ ° i' ZOU Oh ""aOH N\/O 110 NrC N-CH'615. 13 3. 8 0. 6 o Ci OH 'OH No STRUCTURE M. W. IC5o (. M) chlor Nô n CH3 F 111 HCooNt v oF 587. 62 7. 2 0. 8 0 F ° y OH OU oh N N HC N'NNY""-690. 80 4. 1 0. 8 v OH zou "ah 'Y° y F i 113 H3CIN'IN : Na 565. 57 7. 3 1. 1 ,, o F OF Ici Chu v oh hW N i 0 114 HC-, N'IN 588. 67 0. 4 0. 04 0 OH 0 i v'OH No STRUCTURE M. W. tCspM) N/0 N 0 115'C-, -N-N N 588. 67 0. 8 , N/ OH Y° N1 0 oH 116 3 k N 3 570. 69 8. 0 0. 7 N0 I/CHa 0 ", ah xi y 0 117 H3CN ; NJ 598. 70 6. 9 0. 6 XI 'OU 0 ", ah 118 : ;'NJ 622 72 0. 8 0. 1 o if oh No STRUCTURE M. W. Chimp \ N/O 'Y N 119 \^ ''N I 551. 60 8. 8 1. 3 0 OU N 0 I ° oH w I w 0 R <1 ? ° f 640. 78 34. 4 4. 9 N CH, OH ZOU y NN, N \ 121 121 H3c N'IN N 578. 67 3. 0 0. 4 0 ZOU OU N 0 /oh N ° y zou 122"3NNN I I 592. 70 2. 1 0. 4 /nez OH O OH No STRUCTURE M. W. IC N0 ChlRI k NJ 612. 73 11. 7 11. 7 _ 1. 0 OH yin 'oh OH \ N/0 my 124 H3C"'--'N"N _--, NJ 626. 75 6. 4 0. 4 Nez zou pH /' , aOH N/0 Cl, 125 mN ot ° z CH3 605. 14 9. 8 0. 7 o ci ° Sci "OH nez N/O I 126 '\Nw CH 619. 16 10. 3 1. 5 1, M : 0 cl 'OU zoom al. 1 127 H2C. t'N' 624. 74 1. 8 0. 2 N 0 'oh No STRUCTURE M. W. IC50 (UM) C C"", N 0 i \ 128 Ns 590. 68 0. 4 0. 1 N 0 0 i\ OU Chlml N.. 0 r c 129 H2 C N, CF 617. 15 2. 4 0. 5 ci 0 , aOH ni N 0 130 H3CuN5 Nt 642. 75 6. 1 0. 4 II o o 0 neo \ N\/0 'Y L t 4SJ NC 666. 78 2. 2 0. 3 OU No STRUCTURE M. W. IC50 (NM) NEO N 0 I 132 \ ° 668. 79 2. 3 0. 5 i o zou ", ah 'D) N0 1 r 0 a 133 NO N \ 638. 77 3. 5 0. 7 0 zu OH ni il A ChlZl 134 HC, % ndmf° 636. 75 4. 5 0. 9 N C 'oh OH \ Ny0 135 N--ao 595. 65 2. 4 0. 7 ouzo 0 v'oh NoSTRUCTUREM. W. tCsoM) Chl. 1 N\/O I 136"aN" I N N 580. 65 28. 0 2. 9 N N/ 0 OH chl. 1 OB, OH 137 H2 C'N INN 625. 13 0. 6 0. 1 0 cri ZOU oh C"" Oh Chlml N\/0 138"NNN I 623. 11 1. 0 0. 2 N /cl 0 OH A7rs N0 139"zNo "V ? 0 OH 'oh kOH No STRUCTURE M. W. IC5o (NM) I Po 140 HCowN<CI 657. 17 2. 7 0. 3 aï O v'oh Choral Nu0 fi I 141 NIN 594. 69 1. 8 0. 3 N N 0 ou hW Zou N 142"NNN I w/> 596. 71 1. 6 0. 4 NN O y F Oh Chiml F 143 Y C 575. 61 1. 3 0. 2 4\OH OH ii 0 ON No STRUCTURE M. W. IC50 (M) 'Y N 0 . fez F 144 % zX f3-J% 573. 60 2. 1 0. 2 0 F -OH Ti 0 i OH C"" HaW N 145 C'--a 1 610. 71 0. 3 0. 04 N 0 ou hro 0 z "-aOH N\/O 146 I w ° 608. 70 16. 7 1. 4 N N 0 Lo, OH C"" N y0 H2 N 147 C'N 610. 71 9. 4 1. 0 Nu, OH 0 OH l mh, lCso (pM) Chfot N0 N, 148 H3C'0 627. 14 2. 6 0. 3 cl zou OU Oh Chlr N/O . Y I w 149 I 639. 15 31. 0 6. 4 N OH nez OU al. 1 f 150 NY 596. 68 12. 7 0. 7 ° Sr) OH 'OH cnim N0 0 151 o c » ? s ! 596. 68 9. 2 0. 1 0 OU 'oh No STRUCTU RE M sW IC50 (ZM) A ChiMl 152"N'N \ N 622. 72 1. 2 0. 3 ko' ° c OH NYO N 1 0 153 NYNY 622. 72 1. 9 0. 3 val OH v' H I 154 ; < Z S 1 608. 74 3. 2 0. 4 'oh OH F F ni C CH, F N O/ 155 H3C NAv 680. 77 30. 5 4. 1 0 O O OH No STRUCTURE M. W. tCsoM) H3C\CH3 Chl F'\. F H3CN I \ H3C F N 0/ 9-NcO f 678. 75 13. 3 1. 6 /N ? f O O OH ANtO F OU 157 H3C N, nNfq 577. 63 4. 2 0. 1 9 NfX F OH 0'aOH lez Oh 'Y o 158"ZNN I w Nc,. i3 610. 71 0. 9 0. 02 'N/ 0 -OH cnm Chlll 'Y F HZCNNN \ 159 vu 602. 64 2. 7 0. 2 O F 0 N No STRUCTURE M. W. IC50 (M) I Chiât y F i H3C NNnNA 160 9 604. 66 10. 6 0. 5 OF ZON 0 o-n

It has been found according to the present invention that compounds of general formula (1), and especially the compounds of general formula (VI), can inhibit CBP-mediated transcriptional activation in cancer cells due to their specific binding to CBP. This conclusion is supported by immunoprecipitation of CBP of SW480 cells with compounds of the present invention.

The compounds of the present invention can also inhibit the survivin expression in SW480 cells, and therefore, inhibit the oncogenic activity in cancer cells. The compounds of the present invention can be used for inhibiting cancer cells, and thus, would be useful for the regulation of cell growth. Supporting such results, the compounds of the present invention further shows that it can induce the caspase-3 activation in SW480 cells, and therefore, induce the apoptotic activity in cells. The compounds of the present invention can be also advantageously used for inducing apoptosis in cells.

To confirm the oncogenic activity in cancer cell in in vitro MTS cytotoxicity assay was tested by following method.

(1) Cytotoxicity test SW480 or HCT116 cells were placed into 96 well microplate (104cells/well) and incubated for 24 hours at 37 °C. The cells were treated with TCF4 compound at various concentrations for 24 hours. 20 p1 of MTS

solution (Promega) was added into each well and incubated for 2 hours at 37 °C. Cell viability was measured by reading the absorbance at 490nm using microplate reader (Molecular Device) and cytotoxicity of a compound at each concentration was calculated.

(2) Growth Inhibition assay SW480 or HCT116 cells were placed into 96 well microplate (104cells/well) and incubated for 24 hours at 37 °C. 20, ul of [3-(4, 5- dimethylthiazol-2-yl)-5- (3-carboxyphenyl)-2- (4-sulfophenyl)-2H-tetrazolium, inner salt] (MTS) solution (Promega) was added into each well and the absorbance after 2 hour incubation at 37 °C (negative control) was read. And then, the cells were treated with TCF4 compound at various concentrations for 48 hours. 20 pi of MTS solution (Promega) was added into each well and incubated for 2 hour at 37 °C. Cell viability was measured by reading the absorbance at 490nm using a microplate reader (Molecular device) and cytotoxicity of a compound at each concentration was calculated.

The results of oncogenic activity for selected library compounds were shown in the Table 5. The compound numbers is Table 5 are unrelated to the compound numbers in Table 4.

TABLE 5 ONCOGENIC ACTIVITY BY MTS OR SULFORHODAMINE B ASSAY FOR SELECTED LIBRARY COMPOUNDS Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 \ H I i N I O/ . N. I [0 X 2. 28 1. 78 o oH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 XI No oc 2 NNk 2. 58 2. 23 OU Zou N N O i 3 H3CN ; ; S<s 2 73 2 39 0 cri I I a Zu N10 4 H, C, NN N ci 1. 99 1. 91 O cl ZOU zou H F F H'' 'Ny0 0 F 3 E : 2. 32 2. 06 o xi v'oh I F N0 S+F , Y S''F 6 NN N 3. 96 3. 91 ---6 Zou Zou Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 NI N/O F F 'Y F 7"NNY 1. 22 0. 73 zou ,-ah H . N 8 r,, l) <0. 3 <0. 3 li OH oH ()."N, O 9"CCCO 2. 36 1. 92 lof- 0 F F ZION N/0 F F F 10 N"'- (-'-N 2. 34 1. 66 zou o \^ H I NYO X r F 1. 97 1. 30 0"ah o if zou H 12 sNoo 2 54 1 42 ° TU 0-AH If OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 H w I N 1 O F M No 1 65 1. 59 F OH OH O'N y0 NYO 14 \ Na 2. 70 2. 10 ah 'OH . , OH razz / - O /I ON 15 N c 1. 68 1. 34 ci neo NYO O'N If0 N F I NYO 16 \N 5 I F) 4. 18 2. 95 N (S, OF SOH oh NO 17 . Ci 1. 12 0. 74 O F 0 F OH 0Hz neo cl cul 18 N NNCIi3 4. 63 3. 52 N'-O N ins H NYO 19 0 2. 66 1. 17 0-soh xi v'OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 OH / = 0 20 20 5. 02 2. 75 Neo ou- oh = 0 0N N I 21 H3Coof CH3 5. 25 1. 67 CH, N 0 I N 0 HCN CH, . Y 22 HC, N N 6. 58 3. 26 OH ZOU i OH N_r F F 23"9°N-N OH 0 OH 0 No c c, N 24 H3CNN -N \ 13. 79 1. 67 Nu SOH 0 aOH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Xi NO C 25 F6c, NIr cit 24. 53 1. 81 o A 0 C 26 H'°NN - w N' 23. 89 3. 06 o 0 if ni /I N O FI3C CF13 27"N-N 27 H3CN ; F 11. 7 1. 13 N ", ah ni 0"iCCH, 'N' CH'3. 57 5. 47 0 MHz OH tt° sNz 29 X 1a599 793 II NO I/ 0 O Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 nu0 N NO wN 30 sN ; 14. 05 5. 4 Yin ° y ao Chlnl f A\ No N 31 H3CNs 8. 1 + 0. 7 5. 0 1. 0 N 0 : oh Chlhtl CH3 NEZ 0 CI N''CH3 <<E W 47. 2 i 12 16. 9 1. 9 Zozo 0 OU 0 A CH, Chz N/0/ nu0 33 NDupto 28R+20 33 50uM 28. 6 i 2. 0 /nez 0 oh OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT1 16 CH3Chlz' CH, J 34 H, 13. 82. 4 6. 4 6. 4 1. 3 Zon VNa oh O OH mM N 'Y 35 NNN I w " 4. 7 0. 5 5. 0 0. 7 Chlnl 0 0 I 'OH Chine Y 36 %'CH, 21. 9 2. 3 12. 7 1. 3 nez 0 OH CN. 1 /i CH3 CH3 37 NYY 10. 4 0. 8 9. 2 0. 9 OH 0 F 'oh Growth Inhibition Compound Structure (G150, uM) SW480 HCT1 16 Choral N/0 Y CH3 CH3 I 38 H3CoNt< IC\3CH3 8. 5 6. 9 NEZ 0 'OH Chlml N O H3C CH3 y y 39 H, C 4 f N) H 22. 8 i 6. 5 19. 7 3. 3 N 0 OH zou OH N/0 nu0 40 HC, NN'IN No 6. 4+-0. 5 5. 8+-0. 4 ° Sci Zip 'oh Chlrsl CH N J Ha N O 41 HC, N'IN"-N 34. 4 9. 6 14. 7 2. 6 N F OH 0 OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 N 0 C. H3 i H3 42 H3CI N"N N N y CH, 24. 7 10. 8 0 CF6 o OH 'oh Chine 43 H3c,, NDupto 50ut 0 0 CH3 N Ny 0 N O I w 44 N 3. 8 0. 4 4. 2 0. 5 /nez 0 or y I i zu N Chlal N\/0 CH3 F 45 N N 2. 5 0. 2 2. 9 0. 4 N OH 0 OH Growth Inhibition Compound Structure (G15), uM) SW480 HCT116 Chloral N O y-N 46 H, C r'< 5. 5 i 0. 5 9. 2 0. 9 /nez 0 OH ou Chill N/O y 47 H3C, N'INN N 6. 2 12. 2 WOH 0 OH O OH Chl. 1 48 H3CuN rN 20. 7 _ 2. 8 15. 5 2. 3 ° Sci XI 'OU Chu. 1 \ I HzN N/O DX 1. 4iO. 1 1. 0 0. 2 /nez OH ou oh Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Chu. 1 HAN N/0 50 H3NNN I w 4. 6 2. 6 /NEZ po 0 OH \t\OH 0 Ny0 N-N T ruz 51 H3c, N'IN : NTj0 3. 0 0. 1 2. 8 'nez OH 0 OH '\OH N° c N-N 52 VN< 19. 3 2. 1 9. 7 0. 9 /nez N i ( y : : oh \OH 'Y N= : N 53 F Cu f a, N~CH, 11 4 0. 9 4. 7 0. 4 /nez 0 tu 0 OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT1 16 A Chl=l N/O 'Y /> 7. 1 0. 5 4. 9 0. 7 N 0 0 OH phi N/0 cl3 CH3 55 HCo J'4H | 4. 6 i 0. 5 4. 1 0. 7 NEZ 0 OH cnm 56 H3CuN rNuGH3 10. 8 9. 1 N 0 OH criai wOH NINN CH, 0 57 FC, Nr : MN-cH3 3. 1 0. 3 5. 1 0. 3 CI OH ", aOH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Nô N 0 58 H3CIN N N, CN 47. 9 7. 2 22. 3 4. 1 0 cri 'CL OH 'oh wn F aOH | 59 L 55. 1 33. 7 0 y ; 50uM Cl o o" cl criai N/O 0 60 NYr 8. 3 1. 4 6. 3 2. 6 zu ZOU OH H, N han N O 61 11. 3 6. 0 3. 6 0. 3 zon ZIP v'oh Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 ß Chlnl C9 N 0 y N-N 1/cl, -N 62 H3CIN'INN 35. 3 4. 6 23. 5 2. 7 910H 0 0 OH Oh N/O "'N 63 HC, NIINN NH, 18. 8 4. 8 1. 3 0. 1 N O I/i y. b v'oh "-ah /- N N O 64 H3CuNX 12. 0 0. 7 19. 0 1. 6 Neo 0 OH nn cl3 Ha 65 H3C, NN) I--N1 7. 3 4. 7 /NEZ OH 0"aOH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 NH' N. 0/ N-N 66 HCs 3. 0 0. 3 5. 8 0. 3 \t\OH N 0 Oh Oh HZN ^m /\ N 0 67 H3CIN'IN N 0. 6 0. 2 0. 3 i 003 po 0 oh OH N y 0 68 H'NN N \ N 3. 7 0. 2 3. 8 0. 6 IN "'° 'OH \OH 0 xi w A XNt N-N bX/sC 17 9 3. 1 9. 7 1. 0 N I/ N oh I OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Chial 1 NurnO 70 H3CuNv 7. 4 + 0. 6 7. 2 0. 7 nez 0 OU 'T"r' Cl OU I , CH N, 0 71 Y"Y 4. 6 0. 5 3. 6 i 0. 7 OH N 0 (0) 'oh cnm Nst5O C N 72 H3CvNtX 10. 9 0. 6 10. 3 1. 6 o y+, VOX y : : zero OH , , n,. xi N 0 N | 7 N t 9. 2 0. 8 15. 8 2. 6 0 I I N/ O I OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 0 N i0 74 FC, N 0 1. 3 i 0. 4 2. 4 0. 3 N l'), 10 CH3 ", ah v'OH 0 OH Nu0 N 75 HCs9 2. 0 i 0. 1 4. 5 0. 4 0 XI OH o cnm _ T r' N a H'CNNN OH CNa ! O v'oh Chirsl N/0 y F 77 N N 26. 5 6. 5 10. 7 0. 8 N F OH oh OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Nti, Chi. 1 \OH N 0 L zou OH ou ,, cl N 0 y N-N 79 2. 8 0. 2 5. 2 0. 4 'nez I (= \ 'OH N N N 0 80 H3CI N N N 4. 0 0. 6 3. 9 0. 6 u \ cl OH NtO H3CN'CH3 N 81 H \NNN \ 0. 5 0. 3 1. 8 0. 1 zizi CHI a F OU Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 dA Chinì Chifa) N0 H3CN'CH3 HC i 82-N Nr 1. 5 1. 4 O F 0 F OF Oh N0 CH3 83 N)-**N cit 2. 3 0. 3 2. 5 0. 1 zig ICI O 'OH ChWMl CH, F my 84 N N 8. 4 1. 1 9. 9 1. 0 N NtO N 0 OH ni Ny I 85 HCN IN rN 1. 4 0. 5 2. 7 0. 3 0 ° y OH OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Chl. 1 N\/O y F 86 H3C, N'IN N'N, 9. 6 1. 6 6. 5 0. 6 0 F /F HO OH /Chiml N 0 N, . Y I w 87 HCow\N X 0. 6 0. 2 0. 5 0. 1 . nez 0 0 OH Choral N 0 \ 66 HC\N X 0. 3 0. 4 . nez 0 OH Chlml Chu. 1 Ny 0 89 L XN 14. 6 i 1. 4 7. 5 1. 0 I i lez OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Chl l N/0 1 r 0 90 HC, N'INNN12. 6 0. 9 14. 7 1. 0 NEZ 0 zou OH ho 91"No NJ + + 91 HCt=NJ 1. 5 0. 1 3. 2 i 0. 2 OH ", ah chu, l Zou Chlnl 1 N N 0 92 0."N"IN 12. 9 1. 0 14. 9 2. 2 'nez OH Oh Chlal 'IN 11 93 HCNN 1. 9 0. 4 1. 1 0. 1 "-a 11 Neo OH I\ oh Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 CM neo . Y I w 94 NN N 1. 1 0. 3 0. 7 0. 07 N I/ N 0 riz oh OH Chlnl 95 HGNIIN'*, N N 16. 2 2. 6 7. 1 1. 2 NEZ 0 OH ", aOH nu0 O OH o nez Zou cnii N/O y 1 97 H3C F, NJ 3. 7 i 0. 4 3. 4 i 0. 4 o ci o wbt, ci SOH Growth inhibition Compound Structure (G150, uM) SW480 HCT116 I N. 0 CH, 98 H'CNNN I w NcH, 7. 0 1. 1 4. 4 0. 5 cul 0 SOH \ N 9 9 N N 1. 0 0. 05 0. 7 0. 1 0 ZOU Chlrai N 0 100 N'IN N 0. 3 0. 03 0. 4 0. 1 'nez O tir o I OH room OH IN 101"ZNN I w NcH, 1. 1 0. 07 0. 9 0. 1 Y.. Tmcl,- o zizi OH Growth inhibition Compound Structure (G150, uM) SW480 HCT116 am N0 NtO 102 H3CIIN. NN 0 2. 5 0. 4 4. 9 1. 2 N 0 ° su SOH from OH N/0 Nu0 103 HCR N> °Ntl 1. 1 0. 1 1. 5 0. 2 fuzz ° y 0 I OH OVIN N 0 i 104 H2C</\Nv Nt <0. 4 <0. 4 om , o OH xi zou m \ O 105"No w J 2. 8 0. 2 + 2. 1 _ 0. 3 OH O 'OH Growth Inhibition Compound Structure (Gl5 °'uM) SW480 HCT1 16 I N 106 NN 4. 5 0. 3 2. 8 0. 4 N v'/O 'CHs 0 cl, zip 'oh zum nu0 HC,, 107 N'IN'N0 1. 6 0. 1 1. 6 0. 1 ouzo 0 'OU cniri Zu Ny0 I H2-N N 24. 9 2. 2 37. 9 5. 7 N zou O \ oh Chlrsl N/0 109 H2c, N'IN 1. 3 0. 3 1. 1 0. 1 N 1 1 0 ci o OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Chipai 'Y° N 0 k, X51 | 110 N N 2. 1 0. 3 1. 9 0. 1 At° ci OH OU Oh N0 OH po 2. 7 0. 8 'cri 0 zip 'oh airez ex 0 112 N'INN J 5. 1 0. 5 4. 7 0. 3 CI zu 'oh cam Nu0 'ion 6. 8 1. 4 3. 7 0. 6 Un ZOU ou Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 C"" Nu0 114 H2C'1. 7 0. 7 1. 9 0. 2 0 N 0 '\C-OH A LA N 0 F y 115 H2CvN+'F 2. 0 0. 7 1. 1 0. 04 Nr OF ICI 0 SOH A Chl, l N\/0 . fez F i 116 N N 2. 8 0. 9 1. 7 0. 1 c /F OH ON OH \ N/0 H3CN^ 117 C'N 0. 6 0. 1 0. 3 0. 02 1- zou 0 SOH Growth Inhibition Compound Structure (Gl5 0, uM) SW480 HCT116 M1. 1 y0 I 118 % >, N t g 0 N il Zou OH c"" Nu0 fi I 119 Ht NX °) 10. 0 1. 3 9. 5 1. 1 Zon 0 N SOH 'Y° N N\/O . Y I w 120 3 l0e < 1. 8 0. 2 2. 6 0. 1 0 cri OH OU y 0 I 21"D] ;) 6. 2 + 0. 5 13. 1 0. 6 o 0 'OU Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 Y r) NEZ zu f N 122 H3CuNt NX 15. 9 5. 2 14. 8 1. 3 Ao ° ° Sci OH "Y r) N 0 N/O My 123 C' N N 1. 1 0. 3 1. 7 0. 3 Y N z ' H 'aOH 'Y 124 H2C<No NX 2. 3 + 0. 2 1. 4 0. 1 wu 0 OH ol. 1 T ruz 125 2. 2 0. 3 1. 9 0. 2 0 'OH Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 H3C CH3 Chlml r F F -No¢O t r' F 126 H2C<wN, 19. 4 3. 0 11. 6 3. 0 ) a N 0 : oh Chlai Oh 'Y F 127 H3NNN I w 4. 9 0. 7 4. 3 0. 7 ", o F 0 OH n Chl. l < N osa 128 NNYcH, 0. 9 0. 1 1. 0 0. 03 N N 'oh unit ", aOH chill y F 129 ! T t ! 2. 9 0. 5 3. 1 0. 3 \N ZON I\ N Wo Growth Inhibition Compound Structure (G150, uM) SW480 HCT116 A Chl. l Chin) N F i 130 ! i jazz 17. 3 + 1. 2 10. 71. 7 O F ZON 0 I/ N O

In other aspects the present invention provides pharmaceutical compositions containing a compound having the general formula (I), or the general formula (II), or the general formula (III), or the general formula (IV), or the general formula (VI). These compositions may be used in various methods (e. g., treating cancer or Alzheimer's disease) of the present invention as described in detail below.

The pharmaceutical composition of the present invention is formulated to be compatible with its intended route of administration.

Examples of routes of administration include parenteral, e. g. , intravenous, intradermal, subcutaneous, oral (e. g., inhalation), transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents ; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite ; cheating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. In addition, pH may be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be

enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor ELM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound (e. g. , a compound having general formula (I), (II), (III), (IV), or (VI) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.

Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a dispersion medium and the required other

ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile- filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets.

For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutical compatible binding agents, and/or adjuvant materials can be included as part of the composition.

The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin ; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser that contains a suitable propellant, e. g. , a gas such as carbon dioxide, or a nebulizer.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through

the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e. g. , with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutical acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U. S.

Patent No. 4,522, 811.

It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e. g. , for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutical effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50.

Compounds that exhibit large therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i. e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.

For instance, in certain embodiments, a pharmaceutical composition of the present invention is one suitable for oral administration in unit dosage form such as a tablet or capsule that contains from about 1 mg to about 1g of the compound of this invention. In some other embodiments, a pharmaceutical composition of the present invention is one suitable for intravenous, subcutaneous or intramuscular injection. A patient may receive,

for example, an intravenous, subcutaneous or intramuscular dose of about 1 ug/kg to about 1g/kg of the compound of the present invention. The intravenous, subcutaneous and intramuscular dose may be given by means of a bolus injection or by continuous infusion over a period of time. Alternatively a patient will receive a daily oral dose approximately equivalent to the daily parenteral dose, the composition being administered 1 to 4 times per day.

The following table illustrates representative pharmaceutical dosage forms containing the compound or pharmaceutically-acceptable salt thereof for therapeutics or prophylactic use in humans: Tablet 1 mg/tablet Compound 100 Lactose Ph. Eur. 179 Croscarmellose sodium 12.0 Polyvinylpyrrolidone 6 Magnesium stearate 3. 0 Tablet 2 mg/tablet Compound 50 Lactose Ph. Eur. 229 Croscarmellose sodium 12.0 Polyvinylpyrrolidone 6 Magnesium stearate 3. 0 Tablet 3 mg/tablet Compound 1. 0 Lactose Ph. Eur. 92 Croscarmellose sodium 4.0 Polyvinylpyrrolidone 2. 0 Magnesium stearate 1. 0 Capsule mg/capsule Compound 10 Lactose Ph. Eur. 389 Croscarmellose sodium 100 Magnesium stearate 1. 0 Injection I (50mg/ml) Compound 0. 5% w/v Isotonic aqueous solution to 100%

The pharmaceutical composition containing the compound of general formulae (I) or (II) or (III) or (IV) or (VI) can be used for treatment of disorders modulated by Wnt signaling pathway, especially cancer, more especially colorectal cancer.

In one aspect, the present invention provides compounds that inhibit the binding of a radiolabeled enkephalin derivative to the 8 and u opiate receptors. Accordingly, the reverse-turn mimetics of the present invention may be used as receptor agonists and as potential analgesic agents.

In another aspect, the present invention provides methods for inhibiting tumor growth. Such methods comprise the step of administering to a subject (e. g. , a mammalian subject) having a tumor a compound with general formula (I), especially general formula (VI) in an amount effective to inhibit tumor growth. A compound or composition inhibits tumor growth if the tumor sizes are statistically significantly smaller in subjects with the treatment of the compound or composition than those without the treatment.

The inhibitory effect of a particular compound or composition of the present invention on tumor growth may be characterized by any appropriate methods known in the art. For instance, the effect of the compound or composition on survivin expression may be measured. Compounds or compositions down-regulate survivin expression are likely to have inhibitory effects on tumor growth. In addition, assays using tumor cell lines (e. g. , soft agar assays using SW480 cells) and animal models for tumor growth (e. g., nude mice grafted with tumor cells and Min mouse model) may also be used to evaluate the inhibitory effect on tumor growth of a given compound or composition as described in detail in the examples. Other exemplary animal models or xenografts for tumor growth include those for breast cancer (Guo et al., Cancer Res. 62: 4678-84,2002 ; Lu et al., Breast Cancer Res. Treat 57: 183-92,1999), pancreatic cancer (Bouvet et al., Cancer Res. 62: 1534-40, 2002), ovarian tumor (Nilsson et al., Cancer Chemother. Pharmacol. 49: 93-

100,2002 ; Bao etal., Gynecol. Oncol. 78: 373-9,2000), melanoma (Demidem et al., Cancer Res. 61 : 2294-300, 2001), colorectal cancer (Brown eta/., Dig.

Dis. Sci. 45: 1578-84,2000 ; Tsunoda et a/., Anticancer Res. 19 : 1149-52,1999 ; Cao et al., Clin. Cancer Res. 5: 267-74,1999 ; Shawler et al., J. Immunother.

Emphasis Tumor Immunol. 17 : 201-8,1995 ; McGregoreta/., Dis. Colon.

Rectum. 36: 834-9,1993 ; Verstijnen et al., Anticancer Res. 8: 1193-200,1988), hepatocellular cancer (Labonte et al., Hepatol. Res. 18 : 72-85,2000), and gastric cancer (Takahashi et al., Int. J. Cancer 85: 243-7,2000).

The compound or composition that inhibits tumor growth may be administrated into a subject with a tumor via an appropriate route depending on, for example, the tissue in which the tumor resides. The appropriate dosage may be determined using knowledge and techniques known in the art as described above. The effect of the treatment of the compound or composition on tumor growth may also be monitored using methods known in the art. For instance, various methods may be used for monitoring the progression and/or growth of colorectal cancer, including colonoscopy, sigmoidoscopy, biopsy, computed tomograph, ultrasound, magnetic resonance imaging, and positron emission tomography. Methods for monitoring the progression and/or growth of ovarian cancer include, for example, ultrasound, computed tomography, magnetic resonance imaging, chest X-ray, laparoscopy, and tissue sampling.

In a related aspect, the present invention provides a method for treating or preventing cancer. Such methods comprise the step of administering to a subject in need thereof a compound or composition having general formula (I), especially the compound of general formular (VI), in an amount effective to treat or prevent cancer in the subject. Treating cancer is understood to encompass reducing or eliminating cancer progression (e. g., cancer growth and metastasis). Preventing cancer is understood to encompass preventing or delaying the onset of cancer. Various types of cancer may be treated or prevented by the present invention. They include, but are not limited to, lung cancer, breast cancer, colorectal cancer, stomach

cancer, pancreatic cancer, liver cancer, uterus cancer, ovarian cancer, gliomas, melanoma, lymphom, and leukemia.

A subject in need of treatment may be a human or non-human primate or other animal with various types of cancer. A subject in need of prevention may be a human or non-human primate or other animal that is at risk for developing cancer. Methods for diagnosing cancer and screening for individuals with high risk of cancer are known in the art and may be used in the present invention. For instance, colorectal cancer may be diagnosized by fecal occult blood test, sigmoidoscopy, colonoscopy, barium enema with air contrast, and virtual colonoscopy. An individal with high risk of colorectal cancer may have one or more colorectal cancer risk factors such as a strong family history of colorectal cancer or polyps, a known family history of hereditary colorectal cancer syndromes, a personal history of adenomatous polyps, and a personal history of chronic inflammatory bowel disease.

A compound with general formula (I) useful in cancer treatment or prevention may be identified by appropriate methods known in the art.

Methods that may be used to select compounds for inhibitory effect on tumor growth as described above may also be used. The route of administration, the dosage of a given compound, the effectiveness of the treatment may be determined using knowledge and techniques known in the art. Factors that may be considered in making such a determination include, for example, type and stage of the cancer to be treated.

The compound with general formula (I) useful in cancer treatment and prevention may be administered in combination with an anti-neoplastic agent. An anti-neoplastic agent refers to a compound that inhibits tumor growth. Exemplary anti-neoplastic agents include Fluorouracil ; 5-fluor- 2,4 (1 H, 3H) -pyrimidinedione (5-FU), taxol, cisplatin, mitomycin C, tegafur, raltitrexed, capecitabine, and irinotecan (Arango et al., CancerResearch 61, 2001 4910-4915). A compound with general formula (I) administered in combination with an anti-neoplastic agent does not necessarily require that the

compound and the anti-neoplastic agent be administered concurrently. The compound and the agent may be administered separately as long as at a time point, they both have effects on same cancer cells.

In a further related aspect, the present invention provides methods for promoting apoptosis in cancer cells. Such methods comprise the step of contacting cancer cells with a compound having general formula (I), especially a compound having general formula (VI), in an amount effective to promote apoptosis in these cells. A compound promotes apoptosis if the number of cancer cells undergoing apoptosis is statistically significantly larger in the presence of the compound than that in the absence of the compound.

Such compounds may be identified by methods known in the art (e. g., measuring caspase activities and/or cell death) using cultured cancer cell lines, xenografts, or animal cancer models. Preferably, the compound is more active in promoting apoptosis in cancer cells than in normal cells. Cancer cells tratable by the present method may be from various tissue origins.

In another aspect of the present invention, a method for treating a disorder modulated by Wnt signaling pathway in which the method comprises administering to a patient a safe and effective amount of the compounds having general formula (I), especially the compound of general formula (VI) is disclosed. Pharmaceutical composition containing the compound of the present invention can be also used for this purpose. In this connection, it is found in the present invention that the compounds having general formula (I), especially the compound of general formula (VI) or the pharmaceutical composition containing thereof can be useful for the treatment of disorder modulated by TCF4-p catenin-CBP complex, which is believed to be responsible for initiating the overexpression of cancer cells related to Wnt signaling pathway. Thus, it is another aspect of the present invention to provide a method for the treatment of disorder modulated by TCF4-, catenin- CBP complex, using the compounds having the general formula (I), especially the compound of general formula (VI).

The present invention also provides compounds and methods for inhibiting survivin expression. Survivin is a target gene of the TCF/beta- catenin pathway, and more specifically is a target gene of the TCF/beta- catenin/CBP pathway. It is a member of the IAP (Inhibitor of Apoptosis Protein) family of proteins. Biological activity associated with survivin includes: highly expressed at G2/M, regulating cell cycle entry and exit; associated with microtubule, centrosomes, centromeres and midbody depending upon the phases of the cell cycle ; and anti-apoptosis via interacting directly or indirectly with caspases (e. g. , caspase 3,7 and 9). In connection with cancer, survivin is widely and highly expressed in tumor cells, but expressed to little or no extent in normal tissue cells. Also, it has been observed that cancer patients whose tumors expressed survivin had a decreased overall survival. Furthermore, the degree of surviving expression has been correlated with other cancer markers, e. g. , Ki67, PNCA, p53, APC, etc.

The effect of a particular compound of the present invention on survivin expression may be characterized by methods known in the art. Such methods include methods for characterizing survivin expression at the transcriptional or translational level. Exemplary methods for characterizing survivin expression at the transcriptional level are: cDNA microarry, reverse transcription-polymerase chain reaction (RT-PCR), chromatin immunoprecipitation (ChIP), and assays for reporter activities driven by survivin promoter. Exemplary methods for characterizing survivin expression at the translational level are: Western blot analysis, immunochemistry and caspase activities. Detailed descriptions of the above exemplary methods may be found in the examples below.

As described above, the present invention provides methods for inhibiting survivin expression. Such methods comprise the step of contacting a survivin-expressing cell with a compound of the present invention in an amount effective to inhibit survivin expression. A compound inhibits survivin expression if survivin expression in a cell is decreased in the presence of the

compound compared to survivin expression in the absence of the compound.

Survivin-expressing cells include tumor cells that express, such as cells in or from lung cancer, breast cancer, stomach cancer, pancreatic cancer, liver cancer, uterus cancer, ovarian cancer, gliomas, melanoma, colorectal cancer, lymphom and leukemia. The step of contacting the survivin-expressing cells with the compound may be performed in vitro, ex vivo, or in vivo. A compound useful in inhibiting survivin expression may be identified, and the effects of a particular compound of the present invention may be characterized, by appropriate methods known in the art, as described in detail above.

Compounds of the present invention have been shown to inhibit the expression of survivin. Blanc-Brude et al., Nat Medicine 8: 987 (2002), have shown that survivin is a critical regulator of smooth muscle cell apoptosis which is important in pathological vessel-wall remodeling. Accordingly, another aspect of the present invention provides a method of treating or preventing restenosis associated with angioplasty comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic of the present invention. In one embodiment the invention treats the restenosis, i. e., administration of a reverse-turn mimetic of the present invention to a subject having restenosis achieves a reduction in the severity, extent, or degree, etc. of the restenosis. In another embodiment the invention prevents the restenosis, i e., administration of a reverse-turn mimetic of the present invention to a subject that is anticipated to develop new or additional restenosis achieves a reduction in the anticipated severity, extent, or degree, etc. of the restenosis. Optionally, the subject is a mammalian subject.

Compounds of the present invention have been shown to inhibit TCF/B-catenin transcription. Rodova et al., J. Biol. Chem. 277: 29577 (2002), have shown that PKD-1 promoter is a target of the B-catenin/TCF pathway.

Accordingly, another aspect of the present invention provides a method of treating or preventing polycystic kidney disease comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic of

the present invention. In one embodiment the invention treats the polycystic kidney disease, i. e., administration of a reverse-turn mimetic of the present invention to a subject having polycystic kidney disease achieves a reduction in the severity, extent, or degree, etc. of the polycystic kidney disease. In another embodiment the invention prevents polycystic kidney disease, i. e., administration of a reverse-turn mimetic of the present invention to a subject that is anticipated to develop new or additional polycystic kidney disease achieves a reduction in the anticipated severity, extent, or degree, etc. of the polycystic kidney disease. Optionally, the subject is a mammalian subject.

Compounds of the present invention have been shown to inhibit the expression of Wnt signaling. Hanai et al., J. Cell Bio. 158: 529 (2002), have shown that endostatin, a known anti-angiogenic factor, inhibits Wnt signaling.

Accordingly, another aspect of the present invention provides a method of treating or preventing aberrant angiogenesis disease comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic of the present invention. In one embodiment the invention treats the aberrant angiogenesis disease, i. e., administration of a reverse-turn mimetic of the present invention to a subject having aberrant angiogenesis disease achieves a reduction in the severity, extent, or degree, etc. of the aberrant angiogenesis disease. In another embodiment the invention prevents aberrant angiogenesis disease, i. e., administration of a reverse-turn mimetic of the present invention to a subject that is anticipated to develop new or additional aberrant angiogenesis disease achieves a reduction in the anticipated severity, extent, or degree, etc. of the aberrant angiogenesis disease. Optionally, the subject is a mammalian subject.

Compounds of the present invention have been shown to inhibit the expression of Wnt signaling. Sen et al., P. N. A. S. (USA) 97: 2791 (2000), have shown that mammals with rheumatoid arthritis demonstrate increased expression of Wnt and Fz in RA synovial tissue. Accordingly, another aspect of the present invention provides a method of treating or preventing rheumatoid

arthritis disease comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic of the present invention. In one embodiment the invention treats the rheumatoid arthritis disease, i. e., administration of a reverse-turn mimetic of the present invention to a subject having rheumatoid arthritis disease achieves a reduction in the severity, extent, or degree, etc. of the rheumatoid arthritis disease. In another embodiment the invention prevents rheumatoid arthritis disease, i. e., administration of a reverse- turn mimetic of the present invention to a subject that is anticipated to develop new or additional rheumatoid arthritis disease achieves a reduction in the anticipated severity, extent, or degree, etc. of the rheumatoid arthritis disease.

Optionally, the subject is a mammalian subject.

Compounds of the present invention have been shown to inhibit the expression of Wnt signalling. Uthoff et a/., Int J. Oncol. 19: 803 (2001), have shown that differential upregulation of disheveled and fz (Wnt pathway molecules) occurs in ulcerative colitis (compared to Chron's disease patients).

Accordingly, another aspect of the present invention provides a method of treating or preventing ulcerative colitis comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic the present invention. In one embodiment the invention treats the ulcerative colitis, i. e., administration of a reverse-turn mimetic of the present invention to a subject having ulcerative colitis achieves a reduction in the severity, extent, or degree, etc. of the ulcerative colitis. In another embodiment the invention prevents ulcerative colitis, i. e., administration of a reverse-turn mimetic of the present invention to a subject that is anticipated to develop new or additional ulcerative colitis achieves a reduction in the anticipated severity, extent, or degree, etc. of the ulcerative colitis. Optionally, the subject is a mammalian subject.

Compounds of the present invention have been shown to inhibit Wnt TCF/catenin signaling. Accordingly, another aspect of the invention provides a method of treating or preventing tuberious sclerosis complex (TSC) comprising administering to a subject in need thereof a safe and effective

amount of a reverse-turn mimetic the present invention. Subjects having TSC typically develop multiple focal lesions in the brain, heart, kidney and other tissues (see, e. g. , Gomez, M. R. Brain Dev. 17 (suppl) : 55-57 (1995) ). Studies in mammalian cells have shown that overexpression of TSC1 (which expresses hamartin) and TSC2 (which expresses tuberin) negatively regulates cell proliferation and induces G1/S arrest (see, e. g., Miloloza, A. et al., Hum. Mol.

Genet. 9: 1721-1727 (2000) ). Other studies have shown that hamartin and tuberin function at the level of the (3-catenin degradation complex, and more specifically that these proteins negatively regulate beta-catenin stability and activity by participating in the beta-catenin degradation complex (see, e. g. , Mak, B. C. , et al. J. Biol. Chem. 278 (8): 5947-5951, (2003) ). Beta-catenin is a 95- kDa protein that participates in cell adhesion through its association with members of the membrane-bound cadherin family, and in cell proliferation and differentiation as a key component of the Wnt/Wingless pathway (see, e. g., Daniels, D. L., et al., Trends Biochem. Sci. 26: 672-678 (2001) ). Misregulation of this pathway has been shown to be oncogenic in humans and rodents. The present invention provides compounds that modulate p-catenin activity, and particularly its interactions with other proteins, and accordingly may be used in the treatment of TSC. Thus, in one embodiment the invention treats TSC, i. e., administration of a reverse-turn mimetic of the present invention to a subject having TSC achieves a reduction in the severity, extent, or degree, etc. of the TSC. In another embodiment the invention prevents TSC, i. e., administration of a reverse-turn mimetic of the present invention to a subject that is anticipated to develop new or additional TSC achieves a reduction in the anticipated severity, extent, or degree, etc. of the TSC. Optionally, the subject is a mammalian subject.

Compounds of the present invention have been shown to inhibit the expression of Wnt signaling. The Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is expressed in all KSHV-associated tumors, including Kaposi's sarcoma (KS) and p-cei !

malignancies such as primary effusion lymphom (PEL) and multicentric Castleman's disease. Fujimuro, M. et al., Nature Medicine 9 (3): 300-306 (2003), have shown that LANA acts to stabilize P-catenin, apparently by redistribtution of the negative regular GSK-3 ß. The present invention provides compounds and methods for inhibiting p-catenin protein interactions, e. g., ß- catenin/TCF complex formation. Thus, the compounds of the present invention thwart the LANA-induced accumulation of ß-catenin/TCF complex and, at least in part, the consequences of KSHV infection. Accordingly, another aspect of the present invention provides a method of treating or preventing conditions due to infection by Karposi's sarcoma-associated herpesvirus (KSHV). Such conditions include KSHV-associated tumors, including Kaposi's sarcoma (KS) and primary effusion lymphom (PEL). The method comprises administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic the present invention. In one embodiment the invention treats the KSHV-associated tumor, i. e., administration of a reverse-turn mimetic of the present invention to a subject having a KSHV-associated tumor achieves a reduction in the severity, extent, or degree, etc. of the tumor. In another embodiment the invention prevents a KSHV-associated tumor, i. e., administration of a reverse-turn mimetic of the present invention to a subject that is anticipated to develop new or additional KSHV-associated tumors achieves a reduction in the anticipated severity, extent, or degree, etc. of the tumor. Optionally, the subject is a mammalian subject.

LEF/TCF DNA-binding proteins act in concert with activated catenin (the product of Wnt signaling) to transactivate downstream target genes. DasGupta, R. and Fuchs, E. Development 126 (20): 4557-68 (1999) demonstrated the importance of activated LEF/TCF complexes at distinct times in hair development and cycling when changes in cell fate and differentiation commitments take place. Furthermore, in skin morphogenesis, ß-catenin has been shown to be essential for hair follicle formation, its overexpression

causing the"furry"phenotype in mice (Gat, U. , et al. Cell 95 : 605-614 (1998) and Fuchs, E. Harvey Lect. 94: 47-48 (1999). See also Xia, X. et al. Proc. Natl.

Aad. Sci. USA 98 : 10863-10868 (2001). Compounds of the present invention have been shown to inhibit the expression of Wnt signaling, and interfere with formation of ß-catenin complexes. Accordingly, the present invention provides a method for modulating hair growth comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic the present invention, where the amount is effective to modulate hair growth in the subject.

Optionally, the subject is a mammalian subject.

The present invention provides compounds useful in treating or preventing Alzheimer's disease. Alzheimer's disease (AD) is a neurodegenerative disease with progressive dementia. This disease is accompanied by three main structural changes in the brain, namely, i) intracellular protein deposits (also known as neurofibrillary tangles, or NFT), ii) extracellular protein deposits termed amyloid plaques that are surrounded by dystrophic neuritis, and iii) diffuse loss of neurons.

The compounds or compositions of the present invention rescue defectes in neuronal differentiation caused by a presenilin-1 mutation and may decrease the number, or rate at which neuronal precursor populations differentiate to neurons in Alzheimer's brains. Presenilins are transmembrane proteins whose functions are related to trafficking, turnover and cleavage of Notch and Amyloid Precursor Protein. Missense mutations in presenilin 1 (PS- 1) are associated with early-onset familial Alzheimer's disease (Fraser et a/., Biochem. Soc. Symp. 67, 89 (2001) ). The compounds of the present invention may be applicable not only to individuals with PS-1 familial Alzheimer's mutations, but also to general Alzheimer's patients.

In addition, the present invention provides a method for treating or preventing Alzheimer's disease comprising administering to a subject in need thereof a safe and effective amount of a reverse-turn mimetic of the present invention, where the amount is effective to treat or prevent Alzheimer's disease

in the subject. Treating Alzheimer's disease is understood to encompass reducing or eliminating the manifestation of symptoms characteistic of Alzheimer's disease, or delaying the progression of this disease. Preventing Alzheimer's disease is understood to encompass preventing or delaying the onset of this disease.

A subject in need of treatment may be a human or non-human primate or other animal that is at various stages of Alzheimer's disease.

Methods for diagnosing Alzheimer's disese are known in the art (see, e. g., Dinsmore, J. Am. Osteopath. Assoc. 99 (9 Suppl.) : S1-6, 1999; Kurz et al., J.

Neural Transm. Suppl. 62: 127-33,2002 ; Storey et al., Front Viosci. 7: e155-84, 2002; Marin et al., Geriatrics 57: 36-40,2002 ; Kril and Halliday, Int Rev.

Neurobiol. 48: 167-217,2001 ; Gurwitz, Trends Neurosci. 23: 386, 2000; Muller- Spahn and Hock, Eur. Arch. Psychiatry Clin. Neurosci. 249 Suppl. 3: 37-42; Fox and Rossor, Rev. Neuro. (Paris) 155 Suppl. 4: S33-7,1999), including the use of neuropsychological measures, functional imaging measures, biological markers, and autopsy of brain tissue. A subject in need of prevention may be a human or non-human primate or other animal that is at risk for developing Alzheimer's disease, such as an individual having a mutation of certain genes responsible for this disease (e. g. , genes encoding amyloid precursor protein, presenilin 1, and presenilin 2), and/or a gene involved in the pathogenesis of this disease (e. g., apolipoprotein E gene) (Rocchi et al., Brain Res. Bull. 61 : 1- 24,2003).

Compounds with structures as set forth in formula (I) may be screened for their activities in treating or preventing Alzheimer's disease by any appropriate methods known in the art. Such screening may be initially performed using in vitro cultured cells (e. g, PC-12 cells as described in Example 8). Compounds capable of rescuing defects in neuronal differentiation caused by a presenilin 1 mutation may be further screened using various animal models for Alzheimer's disease. Alternatively, compounds with structures as set forth in formula (I) may be directed tested in animal models

for Alzheimer's disease. Many model systems are known in the art and may be used in the present invention (see, e. g., Rowan et al., Philos. Trans. R. Soc.

Lond. B. Biol. Sci. 358: 821-8,2003 ; Lemere et al., Neurochem. Res. 28: 1017- 27,2003 ; Sant'Angelo et al., Neurochem. Res. 28: 1009-15,2003 ; Weiner Harv. Rev. Psychiatry 4: 306-16,1997). The effects of the selected compounds on treating or preventing Alzheimer's disease may be characterized or monitored by methods known in the art for evaluating the progress of Alzheimer's disease, including those described above for diagnosing this disease.

The present invention also provides methods for promoting neurite outgrowth. Such methods comprise the step of contacting a neuron with a compound according to formula (I) in an amount effective to promote neurite outgrowth. These methods are useful in treating neurodegenerative diseases (e. g., glaucoma, macular degeneration, Parkinson's Disease, and Alzheimer's disease) and injuries to nervous system. A compound promotes neurite outgrowth if the neurite lengths of neurons are statistically significantly longer in the presence of the compound than those in the absence of the compound. Such a compound may be identified using in vitro cultured cells (e. g, PC-12 cells, neuroblastoma B104 cell) (Bitar et al., Cell Tissue Res. 298: 233-42,1999 ; Pellitteri et al., Eur. J. Histochem. 45: 367-76,2001 ; Satoh et al., Biochem. Biophys. Res. Commun. 258: 50-3,1999 ; Hirata and Fujisawa, J.

Neurobiol. 32: 415-25,1997 ; Chauvet et a/., Glia 18 : 211-23,1996 ; Vetter and Bishop, Curr. Biol. 5: 168-78,1994 ; Koo et al., Proc. Natl. Acad. Sci. USA 90: 4748-52,1993 ; Skubitz et al., J. Cell BioL 115 : 1137-48,1991 ; O'Shea et al., Neuron 7: 231-7,1991 ; Rydel and Greene, Proc. Natl. Acad. Sci. USA 85: 1257-61,1988) or using explants (Kato et a/., Brain Res. 31 : 143-7, 1983 ; Vanhems et al., Eur. J. Neurosci. 2: 776-82,1990 ; Carri et al., Int J. Dev.

Neurosci. 12 : 567-78,1994). Contacting a neuron with a compound according to the present invention may be carried out in vitro or in vivo. The resulting treated neuron, if generated in vitro, may be transplanted into a tissue in need

thereof (Lacza et al., Brain Res. Brain Res. Protoc. 11 : 145-54,2003 ; Chu et al., Neurosci. Lett 343 : 129-33,2003 ; Fukunaga et al., Cell Transplant 8 : 435- 41, 1999).

The present invention also provides methods for promoting differentiation of a neural stem cell comprising contacting a neural stem cell with a compound according to formula (I) in an amount effective to promote differentiation of a neural stem cell. Such methods are also useful in treating neurodegenerative diseases (e. g., glaucoma, macular degeneration, Parkinson's Disease, and Alzheimer's disease) and injuries to nervous system.

"Neural stem cell"refers to a clonogenic, undifferentiated, multipotent cell capable of differentiating into a neuron, an astrocyte or an oligodendrocyte under appropriate conditions. A compound promotes differentiation of neural stem cells if neural stem cells exhibit a statistically significantly higher degree of differentiation in the presence of the compound than in the absence of the compound. Such a compound may be identified using assays involving in vitro cultured stem cells or animal models (Albranches et al., Biotechnol. Lett. 25: 725-30,2003 ; Deng et al., Exp. Neurol. 182 : 373-82,2003 ; Munoz-Elias et al., Stem Cells 21 : 437-48,2003 ; Kudo et al., Biochem. Pharmacol. 66: 289-95, 2003; Wan et al., Chin. Med. J. 116 : 428-31,2003 ; Kawamorita et al., Hum.

Cell 15 : 178-82,2002 ; Stavridis and Smith, Biochem. Soc. Trans. 31 : 45-9, 2003; Pachernik et al., Reprod. Nutr. Dev. 42: 317-26,2002 ; Fukunaga et al., supra). The neural stem cell may be a cultured stem cell, a stem cell freshly isolated from its source tissue, or a stem cell within its source organism. Thus, contacting the neural stem cell with a compound according to the present invention may be carried out either in vitro (for a cultured or freshly isolated stem cell) or in vivo (for a stem cell within its source organism). The resulting differentiated neural cell, if generated in vitro, may be transplanted into a tissue in need thereof (Lacza et al., supra ; Chu et al., supra; Fukunaga et al., supra).

Such a tissue includes a brain tissue or other nervous tissue that suffers from a trauma or a neurodegenerative disease.

The following non-limiting examples illustrate the compounds, compositions, and methods of use of this invention.

EXAMPLES PREPARATION EXAMPLE 1 PREPARATION OF (N-FMOC-N'-R3-HYDRAZINO)-ACETIC ACID (1) Preparation of N-Fmoc-NV-Methyl Hydrazine 2 L, two-neck, round-bottomed-flask was fitted with a glass stopper and a calcium tube. A solution of methylhydrazine sulfate (20 g, 139 mmol, where R3 is methyl) in THF (300 mL) was added and a solution of DiBoc (33 g, 153 mmol) in THF was added. Saturated sodium bicarbonate aqueous solution (500mL) was added dropwise via addition funnel over 2 hours with vigorous stirring. After 6 hours, a solution of Fmoc-CI (39 g, 153 mmol) in THF was added slowly. The resulting suspension was stirred for 6 hours at 0°C.

The mixture was extracted with ethyl acetate (EA, 500 mL) and the organic layer was retained. The solution was dried with sodium sulfate and evaporated in vacuo. The next step proceeded without purification.

A 1 L, two-necked, round-bottom-flask was fitted with) a glass stopper and a calcium tube. A solution of the product from the previous step in MeOH (300mL) was added and conc. HCI (30 mL, 12 N) was added slowly via addition funnel with magnetic stirring in ice water bath and stirred overnight.

The mixture was extracted with EA (1000 mL) and the organic layer was retained. The solution was dried with sodium sulfate and evaporated in vacuo.

The residue was purified by recrystallization with n-hexane and EA to give N- Fmoc-N'-methyl hydrazine (32.2 g, 83 %). HNMR (DMSO-D6) 8 7. 90-7. 88 (d, J=6 Hz, 2H,), 8 7. 73-7. 70 (d, J=9 Hz, 2H, ), 7. 44-7. 31 (m, 4H), 4. 52-4. 50 (d, J=6 Hz, 2H), 4. 31~4. 26 (t, J=6 Hz, 1H), 2.69 (s, 1H).

(2) Preparation of (N-Fmoc-N'-methyl-hydrazino)-acetic acid t-butyl ester

1 L, two-necked, round-bottom-flask was fitted with a glass stopper and reflux condenser connected to a calcium tube. A solution of N- Fmoc-N'-methyl hydrazine (20 g, 75 mmol) in toluene (300 mL) was added. A solution of t-butylbromo acetate (22 g, 111 mmol) in toluene (50 mL) was added slowly. Cs2CO3 (49 g, 149 mmol) was added slowly. Nal (11 g, 74 mmol) was added slowly with vigorous stirring. The reaction mixture was stirred at reflux temperature over 1 day. The product mixture was filtered and extracted with EA (500 mL). The solution was dried over sodium sulfate and evaporated in vacuo. The product was purified by chromatography with hexane: EA = 2 : 1 solution to give (N-Fmoc-N'-methyl-hydrazino)-acetic acid t-butyl ester (19. 8 g, 70%).

1H-NMR (CDC13-d) 8 7. 78-7. 75 (d, J=9 Hz, 2H, ), 8 7. 61-7. 59 (d, J=6 Hz, 2H, ), 7. 43-7. 26 (m, 4H), 4. 42-4. 40 (d, J=6 Hz, 2H), 4.23 (b, 1 H), 3.57 (s, 2H), 2.78 (s, 3H), 1.50 (s, 9H).

(3) Preparation of (N-Fmoc-N'-methyl-hydrazino)-acetic acid 1 L, two-neck, round-bottomed-flask was fitted with a glass stopper and reflux condenser connected to a calcium tube. (N-Fmoc-N'- methyl-hydrazino)-acetic acid t-butyl ester (20 g, 52 mmol) was added. A solution of HCI (150 mL, 4 M solution in dioxane) was added slowly with

vigorous stirring in an ice water bath. The reaction mixture was stirred at RT over 1 day. The solution was concentrated completely under reduced pressure at 40°C. A saturated aq. NaHCO3 solution (100 mL) was added and the aqueous layer was washed with diethyl ether (100 mL). Conc. HCI was added dropwise slowly at 0°C (pH 2-3). The mixture was extracted and the organic layer was retained (500 mL, MC). The solution was dried with sodium sulfate and evaporated in vacuo. The residue was purified by recrystallization with n-hexane and ethyl acetate to give (N-Fmoc-N'-methyl-hydrazino)-acetic acid (12 g, 72 %). 1H-NMR (DMSO-d6) õ 12. 38 (s, 1H), 8.56 (b, 1H), 7. 89-7. 86 (d, J=9 Hz, 2H, ), 7. 70-7. 67 (d, J=9 Hz, 2H, ), 7. 43-7. 29 (m, 4H), 4. 29-4. 27 (d, J=6 Hz, 2H), 4. 25-4. 20 (t, J=6 Hz, 1 H), 3.47 (s, 2H), 2.56 (s, 3H).

PREPARATION EXAMPLE 2 PREPARATION OF (N-Moc-N'-R7-HYDRAZINO)-ACETIC ACID (1) Preparation of (N'-Methoxycarbonyl-hydrazino)-acetic acid ethyl ester MOC-NH-NH2 (50g, 0.55 mol) was dissolved in DMF (300mut), and then ethyl bromoacetate (68ml, 0.555 mol) and potassium carbonate (77g, 0. 555mol) were added to the reaction vessel. The mixture was warmed to 50°C for 5 hours. After the reaction was completed, the mixture was filtered, and diluted with EtOAc, and washed with brine (3 times). The crude product was purified by column (eluent : Hex/EtOAc = 4/1) to provide 72 of colorless oil.

(2) [N-R7-N'-methoxycarbonyl-hydrazino]-acetic acid ethyl ester

The ethyl ester (10g, 0.05 mol), potassium carbonate (6.9g, 0. 05mol), and R7-bromide (14.1g, 0. 06mol) were dissolved in DMF (200ml), and The mixture was warmed to 50°C for 5hours. After the reaction was completed, the mixture was filtered, and diluted with EA, and washed with brine (3 times). The crude product was purified by Chromatography (eluent : Hex/EtOAc = 4/1).

(3) [N-R7-N'-methoxycarbonyl-hydrazino]-acetic acid

The alkylated ethyl ester (9.5g, 0. 03mol) was dissolved in THF/water (1/1, ml), and added 2N NaOH (28. 3ml) solution at 0 °C. The mixture was stirred at RT for 2 hours. After the starting ester was not detected on UV, the solution was diluted with EA, then separated. The aqueous layer was acidified to pH 3-4 by 1 N HCI, and the compound was extracted by DCM (3 times). The combined organic layer was dried over MgS04, and evaporated to give a yellow solid.

EXAMPLE 1 0 Men Mf-n H MeO , Bn >-") o N + H Pol-0 Pol-O DMSO py_Q g H FmocNH Me Me MeO Bn MeO ° 1. Piperidine/DMF vNz t 2. HOBT/DIC/DMF P°. HCOOH M o"T N 0 Bn N 0 R. T. X, iin 0 Me Bn O Me HOt ! ! : ° O Me (1) Preparation of Nß-Moc-l\P-benzyl-hydrazinoglycine

This compound was prepared according to literature procedure.

(Cheguillaume et. al., Synlett2000, 3,331) (2) Preparation of 1-Methoxycarbonyl-2, 8-dibenzyl-6-methyl-4, 7-dioxo- hexahydro-pyrazino [2, 1-c] [1,2, 4] triazine Bromoacetal resin (60 mg, 0.98 mmol/g) and a solution of benzyl amine in DMSO (2.5 ml, 2 M) were placed in vial with screw cap. The reaction mixture was shaken at 60 °C using rotating oven [Robbins Scientific] for 12 hours. The resin was collected by filtration, and washed with DMF, then DCM, to provide a first component piece.

A solution of Fmoc-alanine (4 equiv., commercially available, the second component piece), HATU (PerSeptive Biosystems, 4 equiv.), and DIEA (4 equiv. ) in NMP (Advanced ChemTech) was added to the resin. After the reaction mixture was shaken for 4 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

To the resin was added 20% piperidine in DMF. After the reaction mixture was shaken for 8 min at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

A solution of N-Moc-W-benzyl-hydrazinoglycine (4 equiv., compound (3) in preparative example 2, where R7 is benzyl, 3rd component piece), HOBT [Advanced ChemTech] (4 equiv. ), and DIC (4 equiv. ) in DMF was added to the resin prepared above. After the reaction mixture was shaken for 3 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then MeOH. The resin was dried in vacuo at room temperature.

The resin was treated with formic acid (2.5 ml) for 18 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under reduced pressure to give the product as an oil.'H-NMR (400 MHz, CDC13) 8 ppm ; 1.51 (d, 3H), 2.99 (m, 1 H), 3.39 (d, 1 H), 3.69 (m, 1 H), 3.75 (m, 1 H), 3.82 (s, 3H), 4.02 (d, 1 H), 4.24 (d, 1 H), 4.39 (d, 1 H), 4.75 (d, 1 H), 5.14 (q, 1H), 5.58 (dd, 1H), 7.10-7. 38 (m, 10H).

EXAMPLE 2 OMe p. OMe Pol RZ \ I n Rz I. 1. Piperidine/DMF p___li Me FmocNH 2. DIEA/DCM N NH j [o-p o B Bn me 0 O Me O O OMe Cl N, 1 Y R4 H Bn nu Ove 1. Piperidine/DMF 0 POI OMe ___R2 ----- Me H- 2. R3-N=C--O Bn /N NH r. t. I N DIEA/DCM N--'Y o Me 0 H H O R4 ! R4 0 N4 0 Rt Example 2 : R 2=-Bn, R4=-CH3 Example 3 : R 2=-CH3, R4=-CH3

(1) Preparation of N'-Fmoc-N-methyl-hydrazinocarbonyl chloride I °/ v HN 0 Pliosgene ci N'N'ko H _ H O CH2C12-aq. NaHC03

An ice-cooled biphasic mixture of N-methyl hydrazine carboxylic acid 9H-fluoren-9-ylmethyl ester (107 mg, 0.4 mmol) in 15 ml of CH2CI2 and 15 ml of saturated aq. NaHC03 was rapidly stirred while 1.93 M phosgene in toluene (1.03 ml, 2 mmol) was added as a single portion. The reaction mixture was stirred for 30 min, the organic phase was collected, and the aqueous

phase was extracted with CH2CI2. The combined organic layers were dried over MgS04, filtered, and concentrated in vacuo to afford 128 mg (97 %) of carbamoyl chloride as a foamy solid. [Caution: Phosgene vapor is highly toxic.

Use it in a hood]. This product was used for the following solid phase synthesis without further purification.

(2) Preparation of 2, 5-Dimethyl-7-benzyl-3, 6-dioxo-hexahydro- [1,2, 4] triazolo [4, 5-a] pyrazine-1-carboxylic acid benzylamide Bromoacetal resin (30 mg, 0.98 mmol/g) and a solution of benzyl amine in DMSO (1.5 ml, 2 M) were placed in vial with screw cap. The reaction mixture was shaken at 60 °C using rotating oven [Robbins Scientific] for 12 hours. The resin was collected by filtration, and washed with DMF, then DCM, to provide the first component piece.

A solution of Fmoc-alanine (3 equiv. , second component piece, commercially available), HATU (PerSeptive Biosystems, 3 equiv. ), and DIEA (3 equiv. ) in NMP (Advanced ChemTech) was added to the resin. After the reaction mixture was shaken for 4 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF, to thereby add the second component piece to the first component piece.

To the resin was added 20% piperidine in DMF. After the reaction mixture was shaken for 8 min at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

A solution of N'-Fmoc-N-methyl-hydrazinocarbonyl chloride (combined third and fourth component pieces, 5 equiv. ) obtained in the above step (1), DIEA (5 equiv. ) in DCM was added to the resin prepared above. After the reaction mixture was shaken for 4 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and DMF.

To the resin was added 20% piperidine in DMF (10 mi for 1 g of the resin). After the reaction mixture was shaken for 8 min at room

temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

The resin was treated with a mixture of benzyl isocyanate (4 equiv. ) and DIEA (4 equiv. ) in DCM for 4 hours at room temperature. Then, the resin was collected by filteration and washed with DMF, DCM, and then MeOH. The resin was dried in vacuo at room temperature.

The resin was treated with formic acid for 14 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under reduced pressure to give the product as an oil.

1H-NMR (400 MHz, CDCl3) 6 ppm ; 1.48 (d, 3H), 2.98 (s, 3H), 3.18 (m, 1H), 3.46 (m, 1H), 4.37-4. 74 (m, 5H), 5.66 (dd, 1H), 6.18 (m, 1H), 7.10-7. 40 (m, 10H).

EXAMPLE 3 PREPARATION OF 2,5, 7-TRIMETHYL-3, 6-DIOXO-HEXAHYDRO- [1, 2, 4] TRIAZOLO [4, 5- A] PYRAZINE-1-CARBOXYLIC ACID BENZYLAMIDE The title compound is prepared according to the same procedure as described in Example 2, but reacting bromoacetal resin with a solution of methyl amine instead of benzyl amine. 1H-NMR (400 MHz, CDC13) 6 ppm ; 1.48 (d, 3H), 2.99 (s, 3H), 3.03 (s, 3H), 3.38 (m, 1 H), 3.53 (dd, 1 H), 4.36 (dd, 1H), 4.52 (q, 1H), 4.59 (dd, 1H), 5.72 (dd, 1H), 6.19 (br. t, 1H), 7.10-7. 38 (m, 5H).

EXAMPLE 4 PREPARATION OF 2-METHYL-5- (P-HYDROXYPHENYLMETHYL)-7-NAPHTHYLMETHYL- 3, 6-DIOXO-HEXAHYDRO-[1,2,4]TRIAZOLO[4,5-A]PYRAZINE-1-CARBOXYLI C ACID BENZYLAMIDE Bromoacetal resin (30 mg, 0.98 mmol/g) and a solution of naphthylmethyl amine in DMSO (1.5 ml, 2 M) were placed in vial with screw

cap. The reaction mixture was shaken at 60°C using rotating oven [Robbins Scientific] for 12 hours. The resin was collected by filtration, and washed with DMF, then DCM to provide the first component piece.

A solution of Fmoc-Tyr (OBut) -OH (3 equiv. ), HATU (PerSeptive Biosystems, 3 equiv. ), and DIEA (3 equiv. ) in NMP (Advanced ChemTech) was added to the resin. After the reaction mixture was shaken for 4 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF, to thereby add the second component piece to the first component piece.

To the resin was added 20% piperidine in DMF. After the reaction mixture was shaken for 8 min at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

A solution of V-Fmoc-N-methyl-hydrazinocarbonyl chloride (5 equiv. ), DIEA (5 equiv. ) in DCM was added to the resin prepared above. After the reaction mixture was shaken for 4 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and DMF.

To the resin was added 20% piperidine in DMF (10 ml for 1 g of the resin). After the reaction mixture was shaken for 8 min at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

The resin was treated with a mixture of benzyl isocyanate (4 equiv. ) and DIEA (4 equiv. ) in DCM for 4 hours at room temperature. Then, the resin was collected by filteration and washed with DMF, DCM, and then MeOH. The resin was dried in vacuo at room temperature.

The resin was treated with formic acid for 14 hours at room temperature. After the resin was removed by filtration, the filtrate was condensed under reduced pressure to give the product as an oil.

1H-NMR (400 MHz, CDC13) 5 ppm; 2.80-2. 98 (m, 5H), 3.21-3. 37 (m, 2H), 4. 22-4. 52 (m, 2H), 4.59 (t, 1H), 4.71 (d, 1H), 5.02 (dd, 1H), 5.35 (d, 1H), 5.51 (d, 1H), 6.66 (t, 2H), 6.94 (dd, 2H), 7.21-8. 21 (m, 12H).

EXAMPLE 5 <BR> <BR> <BR> <BR> <BR> <BR> PREPARATION OF 2-METHYL-6- (P-HYDROXYPHENYLMETHYL)-8-NAPHTHYL-4, 7-DIOXO- HEXAHYDRO-PYRAZINO [2, 1-c] [1,2, 4] TRIAZINE-1-CARBOXYLIC ACID BENZYLAMIDE Bromoacetal resin (60 mg, 0.98 mmol/g) and a solution of naphthyl amine in DMSO (2.5 ml, 2 M) were placed in vial with screw cap. The reaction mixture was shaken at 60 °C using rotating oven [Robbins Scientific] for 12 hours. The resin was collected by filtration, and washed with DMF, then DCM.

A solution of Fmoc-Tyr (OBut) -OH (4 equiv. ), HATU [PerSeptive Biosystems] (4 equiv. ), and DIEA (4 equiv. ) in NMP (Advanced ChemTech) was added to the resin. After the reaction mixture was shaken for 4 hours at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

To the resin was added 20% piperidine in DMF. After the reaction mixture was shaken for 8 min at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

A solution of Nß-Fmoc-Na-benzyl-hyrazinoglycine (4 equiv.), HOBT [Advanced ChemTech] (4 equiv. ), and DIC (4 equiv. ) in DMF was added to the resin prepared above. After the reaction mixture was shaken for 3 hours at room temperature, the resin was collected by filtration and washed with DMF, and then DCM. To the resin was added 20% piperidine in DMF (10 ml for 1 g of the resin). After the reaction mixture was shaken for 8 min at room temperature, the resin was collected by filtration and washed with DMF, DCM, and then DMF.

The resin was treated with a mixture of benzyl isocyanate (4 equiv. ) and DIEA (4 equiv. ) in DCM for 4 hours at room temperature. Then, the resin was collected by filteration and washed with DMF, DCM, and then MeOH. After the resin was dried in vacuo at room temperatur, the resin was treated with formic acid (2.5 ml) for 18 hours at room temperature. The resin

was removed by filtration, and the filtrate was condensed under reduced pressure to give the product as an oil.

'H-NMR (400 MHz, CDCl3) # ppm; 2.73 (s, 3H), 3.13 (d, 1H), 3.21-3. 38 (m, 3H), 3.55 (d, 1 H), 3.75 (t, 1 H), 4.22 (dd, 1 H), 4.36 (dd, 1 H), 4.79 (d, 1H), 5.22 (t, 1H), 5.47 (m, 2H), 6.68 (d, 2H), 6.99 (d, 2H), 7.21-8. 21 (m, 12H); MS (m/z, ESI) 564.1 (MH+) 586.3 (MNa+).

EXAMPLE 6 BIOASSAY FOR THE MEASUREMENT OF C50 AGAINST SW480 CELLS AND CYTOTOXICITY TEST ON THE CELL LINES The test compound (Compound A) used in this example was prepared in Example 4. a. Reporter Gene Assay SW480 cells were transfected with the usage of Superfect transfect reagent (Qiagen, 301307). Cells were trypsinized briefly 1 day before transfection and plated on 6 well plate (5 x 105 cells/well) so that they were 50-80% confluent on the day of transfection.

Four microgram (TOPFlash) and one microgram (pRL-null) of DNAs were diluted in 150 lli of serum-free medium, and 30 aul of Superfect transfect reagent was added. The DNA-Superfect mixture was incubated at room temperature for 15 min, and then, 1 ml of 10 % FBS DMEM was added to this complex for an additional 3 hours of incubation. While complexes were forming, cells were washed with PBS twice without antibiotics.

The DNA-Superfect transfect reagent complexes were applied to the cells before incubating at 37 °C at 5 % CO2 for 3 hours. After incubation, recovery medium with 10 % FBS was added to bring the final volume to 1.18 ml. After 3 hours incubation, the cells were harvested and reseeded to 96 well plate (3 x 104 cells/well). After overnight incubation at 37 °C at 5 % COs, the cells were treated with Compound A for 24 hours. Finally, the activity was checked by means of luciferase assay (Promega, E1960).

Figure 3 illustrates the results of the measurement of IC50 of Compound A for SW480 cells. b. Sulforhodamine B (SRB) assay Growth inhibitory effect of Compound A on the cells listed below was measured by the sulforhodamine B assay. SW480 cells in 100 pi media were plated in each well of 96-well plate and allowed to attach for 24 hours.

Compound A was added to the wells to produce the desired final concentrations, and the plates were incubated at 37 °C for 48 hours. The cells were then fixed by gentle addition of 100 pi of cold (4 °C) 10% trichloroacetic acid to each well, followed by incubation at 4 °C for 1 hour. Plates were washed with deionized water five times and allowed to air dry. The cells were then stained by addition of 100 gul SRB solution (0.4% SRB (w/v) in 1 % acetic acid (v/v) ) to wells for 15 min. After staining, the plates were quickly washed five times with 1 % acetic acid to remove any unbound dye, and allowed to air dry. Bound dye was solubilized with 10 mmol/L Tris base (pH 10.5) prior to reading the plates. The optical density (OD) was read on a plate reader at a wavelength of 515nm with Molecular Device. Inhibition of growth was expressed as relative viability (% of control) and Gl50 was calculated from concentration-response curves after log/probit transformation.

Table 6 shows in vitro cyctotoxicity (SRB) assay data for Compound A obtained in Example 4. The values in Table 6 are in ug/ml.

TABLE 6 Origin Cell Example Cisplatin 5-FU Colon T84 1. 134 > 10 1. 816 LOVO 0. 532 > 10 1. 029 HT29 1. 694 > 10 5. 334 DLD-1 1. 775 > 10 > 10 COL0205 1. 136 > 10 1. 130 CACO-2 1. 201 > 10. 0. 451 SW480-Kribb 1. 137 > 10 > 10 SW480-CWP 0. 980 4. 502 > 10 SW620 1. 426 > 10 5. 570 KM12 1. 451 > 10 2. 729 HCT15 2. 042 > 10 1. 179 HCT116 0. 96 > 10 1. 039 HCC2998 1. 047 > 10 5. 486 786-0 1. 417 3. 347 0. 584 Leukemia HL60 1. 243 > 10 7. 010 RPM18226 1. 1. 177 > 10 > 10 K562NIN 1. 640 > 10 7. 071 K562/ADR 7. 682 > 10 > 10 K562 1. 247 > 10 6. 133 Prostate PC3 1. 207 > 10 > 10 HT1080 1. 469 > 10 0. 798 Lung A549 1. 386 > 10 1. 007 NCI H460 1. 498 > 10 1. 397 NCI H23 1. 296 5. 176 2. 254 Renal 293 0. 731 6. 641 2. 015 CAKI-1 0. 467 > 10 0. 925 ACHN 1. 263 5. 019 5. 062 Melanoma RPM17951 0. 936 5. 010 0. 920 M14 2. 289 3. 447 1. 225 HMV-II 4. 834 3. 190 0. 695 HMV-I 1. 153 5. 478 2. 110 G361 0. 584 4. 827 1. 539 CRL1579 1. 830 0. 699 > 10 A431 1. 083 3. 722 0. 404 A253 1. 398 2. 084 2. 926 UACC62 0. 563 > 10 1. 093 SK-MEL-28 1. 291 > 10 > 10 SK-MEL-5 0. 888 > 10 2. 434 LOX-IMVI 1. 526 > 10 > 10 A375 1. 391 > 10 1. 464 Breast MCF7/ADR 9. 487 9. 907 > 10 MCF7 7. 355 > 10 1. 751

EXAMPLE 7 MIN MOUSE MODEL Selected compounds of the present invention (Compound B and Compound C) were evaluated in the min mouse model to evaluate their efficacy as anit-cancer agents.

Compound B

Compound C The min mouse model is a widely used model to test for this type of efficacy. The numbers of polyp formed in small intestine and colon of these mice after various treatments were measured (Table 7). The data shown that both compounds, when administered at about 300 mpk, reduce the number of polyp in min mice compared to those in the control mice treated with vehicle only.

TABLE 7 MIN MOUSE MODEL DATA Polyp Number (Mean S. S. D.) % P (total) Group Small Colon Total Vs. VH Inhibition Intestine vs. VH Wild Type 0. 00. 0 0. 00. 0 0. 00, 0-- Vehicle 65. 815. 9 1. 8+1. 5 67. 7+15. 3 Compound C 69. 2+20. 8 1. 7+1. 5 71. 4+23. 0 69. 220. 8 1. 71. 5 71. 423. 0-- -100 mpk Compound C 46. 117. 1 1. 11. 2 47. 016. 9 <0. 01 31 - 300 mpk Compound B "45. 222. 1 1. 40. 9 46. 817. 0 <0. 01 31 - 300mpk -300 mpk 48. 020. 7 0. 50. 5 48. 520. 9 <0. 05 28 - 160 ppm

EXAMPLE 8 CHEMOGENOMIC INHIBITION OF CBP/D CATENIN INTERACTION RESCUES DEFECTS IN NEURONAL DIFFERENTIATION CAUSED BY A PRESENILIN-1 MUTATION The following compound (Compound D) was used in this example : Materials and Methods Plasmids. TOPFLASH and FOPFLASH reporter constructs were transformed into DH5a competent cells by standard protocol. Plasmids used

for transfection assays were isolated and purified using EndoFree Maxi Kit (Qiagen, Valencia, CA).

PC-12 Cell Culture. PC-12 cells were maintained in RPMI 1640 supplemented with 10% horse serum, 5% fetal bovine serum, 4.5 g/L glucose, 2 mM L-glutamine, 1.0 mM sodium pyruvate and 10 lug/ml penicillin- streptomycin.

Cell Differentiation. Cell culture dishes were pre-coated overnight with 0.25 mg/ml collagen (Cohesion, CA), 10 ug/ml Poly-L-Lysine (Sigma- Aldrich, St. Louis, MO) and 12 ug/ml Polyethyleneimine (ICN, La Mesa, CA).

Cells were cultured on coated dishes at 15,000 cells/cm2, and differentiated into a neuron-like phenotype by incubation in medium with reduced serum (1 % fetal bovine serum), containing 50 ng/ml nerve growth factor (NGF) (Sigma-Aldrich) for 10 days. NGF-containing medium was changed every 2-3 days.

Treatment with Compound D. Compound D, a small molecule inhibitor of ß-catenin/CBP interaction, was dissolved in DMSO at a stock concentration of 100 mM. Differentiated PC-12/L286V cells were treated with increasing concentrations of this compound for 4 hours. Transfection was then initiated after this treatment period. For cell differentiation experiments, Compound D was added at a concentration of 10 uM, together with NGF, for the entire differentiation period.

Transfection. PC-12 cells were cultured and differentiated on 60- mm dishes. At the end of the 10-day differentiation period, cells were transfected with 2 ug reporter constructs, TOPFLASH and FOPFLASH, per 60- mm dish. Transfections were performed using Superfect (Qiagen) according to manufacturer's instructions.

Luciferase Assays. Cells were lysed, 6 hours after transfections, in 100 ul of Cell Culture Lysis Reagent (Promega, Madison, WI), and scraped into microcentrifuge tubes. Tubes were then centrifuged briefly (about 10 seconds) at 12000 rpm to pellet cell debris. Luciferase activity was measured on 20 ul of cell lysate and 100 ul substrate from the Luciferase Assay System

(Promega). Luciferase activity was measure using Packard LumiCount.

(Hewlett Packard). Quantitation of luciferase was performed in triplicates, and repeated in at least three independent experiments.

Immunofluorescence. Cells were plated at a density of 10,000 cells/cm2 on sterile coated 22x22 mm coverslips in a 6-well culture plate.

Differentiation was initiated, as previously described, for 10 days. The differentiated cells were then fixed in methanol for 15 minutes at-20°C. This is followed by a 15 minutes incubation with PBS + 0. 1% Triton X-100. The coverslips were incubated with antibodies raised against Ephrin B2 Receptor (Santa Cruz Biotechnology) and Gap-43 (Novus Biologicals) for 40 minutes at 37°C. After a series of washes with PBS-Triton X-100, secondary antibody conjugated to FITC (Jackson ImmunoResearch, Westgrove, PA) was applied.

All slides images were acquired using a Nikon PCM2000 Laser Scanning Confocal Microscope mounted on a Nikon Eclipse E600 upright microscope (Nikon, Melville, NY).

Quantitation of Neurite Outgrowth. Cell counts were taken from six randomly chosen microscopic fields (10x). In each field, total number of cells, as well as cells that displayed neurites greater than twice the length of the cell body was determined. The number of cells with such outgrowths was then expressed as a percentage of the total number of cells. Values obtained were from duplicates of three independent experiments.

RT-PCR. To analyze the mRNA levels for Ephrin B2 (EphB2) receptor, total RNA was isolated using Trizol (Invitrogen-GIBCO-BRL, Baltimore, MD) from differentiated ce ! ts. 2 ug RNA was reverse transcribed in a total volume of 20 ul with random hexamer (50 ng), and using the Superscript II reverse transcription system (Invitrogen-GIBCO-BRL), according to manufacturer's guidelines. PCR was carried out in a 50 pi volume containing 5 PI cDNA, 100 pmol primers, 100 uM dNTPs, 1X Taq buffer and 1.5 mM MgC12.

Reaction mixtures were heated to 80°C for 10 min, after which Taq was added. cDNAs were amplified for 25 (EphB2 receptor) or 15 (GAPDH) cycles. One

round of amplification consisted of 1 min at 94°C, 2 min at 60°C, and 2 min at 72°C, with a final extension time of 10 min at 72°C. The PCR products were resolved and visualized by electrophoresis in a 2% gel, stained with ethidium bromide. EphB2 receptor PCR primers used were, 5'- CACTACTGGACCGCACGATAC-3'and 5'- TCTACCGACTGGATCTGGTTCA-3'. Primer pairs for GAPDH were 5'- GGTGCTGAGTATGTCGTGGA-3'and 5'-ACAGTGTTCTGGGTGGCAGT- 3'.

Results Rat PC-12 cells are derived from the neural crest lineage and upon nerve growth factor (NGF) treatment, undergo differentiation to a neurite- bearing sympathetic-like neuron (Greene and Tischler, Proc NatlAcad Sci U S A 73,2424 (1976) ). Utilizing a PC-12 cell based model, the effects of an early- onset FAD associated PS-1 mutation, PS-1/L286V, on TCF/P-catenin mediated transcription and neuronal differentiation were characterized. It has been demonstrated that specifically blocking transcription mediated by TCF/ (3- catenin/CBP alleviates PS-1 induced defects in neuronal differentiation.

PC-12 cells stably overexpressing either wild type PS-1 (PS- 1/WT) or mutant PS-1 (PS-1/L286V) and a vector-transfected control cell line (Guo et al., Neuroreport, 8,379 (1996) ) were plated on dishes coated with collagen, poly-L-lysine and poly-etheleneimine. Differentiation was induced by treatment with 50ng/ml of NGF for 10 days. Overexpressing PS-1/WT cells or the vector-transfected cells had extensive neurite formation (similar to PC-12 cell clones from ATCC), whereas the PS-1/L286V mutant cells had only stubby neurite formation (Fig. 4 A-C). Additionally, vector-transfected PC-12 control and PS-LN\IT cells displayed extensive expression of the neuronal differentiation markerGAP-43 (Gorgels eta/., NeurosciLett. 83,59 (1987) ) (Fig. 4 D, E), whereas the PS-1/L286V cells were essentially devoid of this marker (Fig. 4 F).

To assess the effects of the PS-1/L286V mutation on canonical Wnt/p-catenin signaling, we transiently transfected NGF treated PC-12 cells with Topflash, a Wnt/p-catenin signaling reporter construct (Morin et al., Science 275,1787 (1997) ). As seen in Figure 4F, the overexpressing PS- 1/WT cells had similar levels of TCF/ß-catenin signaling compared to the vector control cells. However, the PS-1/L286V mutant cells displayed significantly (10-fold) increased Topflash expression. In contrast, the negative control reporter construct Fopflash did not show any significant differences.

It was hypothesized that dysregulated TCF/ß-catenin signaling in the PS-1/L286V mutant cells was responsible for the defective differentiation and neurite outgrowth. To test this hypothesis, a specific small molecule inhibitor of TCF/ (3-catenin signaling, Compound D (Emami et al., Cancer Cell, in press), was used. This small molecule selectively blocks the (3-catenin/CBP interaction, but not the ß-catenin/p300 interaction, thereby interrupting a subset of TCF/ (3-catenin transcription. Treatment of the PS-1/L286V mutant cells with 10µM Compound D plus NGF decreased TCF/ß-catenin reporter gene transcription, and led to essentially normal neurite outgrowth and differentiation (Fig. 5 A), similar to that seen in the overexpressing PS-1/WT cells (Figs. 5 A, B), as compared to the untreated cells (Fig. 4 C). Furthermore, PS-1/L286V mutants treated with Compound D showed similar intense GAP-43 staining to the PS-1/WT and vector-transfected cells (Fig. 4 B). To demonstrate that Compound D treated mutant cells develop neurites similar to that of the vector control or PS-1/WT cells, cells that had neurites greater than twice the length of the cell body were counted. Treatment with Compound D substantially increased the percentage of cells bearing neurites to levels similar to that of the vector-transfected and overexpressing PS-1/WT cells (Fig. 5 C). It is concluded that blocking transcription mediated by TCF/ß-catenin/CBP corrects many of the phenotypic defects in neurite outgrowth and neuronal differentiation due to the PS-1/L286V mutation.

Ephrin B2 receptors (EphB2) have been implicated in synapse formation (Wilkinson, Nat. Rev. Neurosci. 2,155 (2001) ) and the Ephrin A family has recently been shown to play a role in hippocampal dendritic spine morphology (Murai et al., Nat. Neurosci. 6,153 (2003) ). Focused EphB2 expression was observed, which localized with neuronal processes in the vector and PS-1/WT-transfected cells (Fig. 6 A, B), whereas the PS-1/L286V mutant cells demonstrated very weak and diffuse EphB2 signal (Fig. 6 C).

Increased TCF/ (3-catenin signaling in PS-1/L286V mutant cells manifested itself in decreased EphB2 expression as judged by RT-PCR (Fig. 6 E, lane 3).

Furthermore, addition of 10uM Compound D led to increased EphB2 message (Fig. 6 E, lane 4) as well as EphB2 expression in these cells (Fig. 6 D). These results are consistent with the data of Batile and colleagues (Batlle et al., Cell 111,251 (2002) ) who recently showed that expression of EphB2/EphB3 receptors and their ligand ephrin-B1 is inversely controlled in colonic crypts via TCF/ß-catenin transcription, and that proper regulation is important for appropriate cell proliferation, differentiation and sorting. We present evidence that the PS-1/L286V mutation via increased TCF/p-catenin signaling, decreased the expression of EphB2 receptors and this is corrected by Compound D mediated inhibition of the ß-catenin/CBP interaction.

EXAMPLE 9 COMPOUND D CAUSES A G1/S-PHASE ARREST AND ACTIVATES CASPASE ACTIVITY Flow Cytometric Analysis (FACS) For FACS analysis, approx. 5 X 106 cells from Compound D- treated or vehicle-treated were fixed with 70% chilled ethanol and stored at-20 °C for at least 30 minutes. The cells were washed once with 1x PBS and incubated with propidium iodine (PI) solution (85, ug/ml propidium iodine, 0. 1% Nonidet P-40,10 mg/ml RNAse) for 30 minutes at room temperature. 10,000 stained cells for each sample were acquired using Beckman Coulter EPICS XL-

MCL Flow Cytometry and the percentage of cells in different phase of the cell cycle was determined by Expo32 ADC software (Coulter Corporation, Miami, Florida, 33196).

Caspase-3 Activity Assay SW480, HCT116, and CCD18Co cells were plated at 105 cells per well (96-well plates) for 24 hours prior to treatment. 25 IlM of Compound D or control (0.5% DMSO) was added to each well. 24 hours post treatment, cells were lysed and caspase activity was measured using a caspase-3/7 activity kit (Apo-One Homogeneous caspase-3/7 assay, #G77905, Promega). Relative fluorescence units (RFU) were obtained by subtracting the unit values of the blank (control, without cells) from the experimental measured values.

Compound D Causes a G1/S-Phase Arrest and Activates Caspase Activity It has been shown that inhibition of the expression of the cyclin D1 gene causes arrest at the G1/S-phase of the cell cycle (Shintani et al., "Infrequent alternations of RB pathway (Rb-p161NK4A-cyclin D1) in adenoid cystic carcinoma of salivary glands,"Anticancer Res. 20: 2169-75 (2000)).

HCT116 (Figure 7A, upper panel) and SW480 (Figure 7A, lower panel) cells were treated with Compound D (25 M) (Figure 7A, right) or control (0.5% DMSO) (Figure 7A, left) for 24 hours. The cells were subsequently stained with propidium iodide (PI) and analyzed for DNA content by FACS cytofluorometry. As expected, the control cells, (Figure 7A, left), were cycling normally whereas the Compound D treated cells (Figure 7A, right) showed increased accumulation at G1/S-phase of the cell cycle. Thus, it can be seen that Compound D causes arrest of cells at the G1 phase.

Caspases are cysteine proteases that are generally activated in a given population of cells triggered by apoptotic stimuli. To assess apoptotic induction in SW480, HCT116, and wild-type colonocytes (CCD18Co cells), the cells were treated with either Compound D (2511M) or control (0.5% DMSO) for

24 hours, followed by an assay for caspase-3/7 activity. As shown in Figure 7B, Compound D specifically and significantly activated the caspase-3/7 pathway in SW480 and HCT116 cells compared to CCD18Co cells.

EXAMPLE 10 COMPOUND D REDUCES PROLIFERATION OF TRANSFORMED COLORECTAL CELLS Soft Agar Assays The soft agar colony formation assay was conducted with SW480 cells by some modification of the procedure previously described (Moody et al., "A vasoactive intestinal peptide antagonist inhibits non-small cell lung cancer growth,"Proc. Natl. Acad. Sci. USA. 90: 4345-49 (1993)).

Each well (35mm) of a 6-well plate (Nalge Nunc International, Roskide, Denmark) was coated with 1ml of 0.8 % bottom agar in DMEM medium containing 10% fetal bovine serum. After it was solidified, 1 ml of DMEM medium containing 0.4 % top agar, 10% fetal bovine serum, compound doubly concentrated, and 5,000 single viable cells was added to each well. The cultures were incubated at 37 °C in humidified 5% C02 incubator. Colonies in soft agar were monitored daily and photographed after incubation for 8 days.

Colonies > 60 µm in diameter were counted.

Compound D Reduces Proliferation of Transformed Colorectal Cells Soft agar colony forming assays were performed using SW480 cells treated with Compound D (0. 25-5 iM) and 5-fluorouracil (5-FU) (0.5-32 pM). As shown in Figure 8A, Compound D shows a dose dependent decrease in the number of colonies formed. IC50 value of Compound D and 5-FU was 0.87 ~ 0. 11 LM and 1.98 0. 17 tM, respectively. Thus, Compound D increased caspase activity and reduced growth in vitro of colorectal cells that are transformed by mutations that activate (3-catenin signaling.

EXAMPLE 11 COMPOUND C REDUCES TUMOR GROWTH IN NUDE MOUSE MODEL SW620 cells (9X106 cells/mouse) were grafted into nude mice subcutaneously on Day 0. Mice received 200 mg/kg of Compound C intraperitoneally every other day until Day 21 after 4 times of 300 mg/kg every other day starting Day 1. Compound C reduces the tumor growth in the treated mice compared to the vehicle control mice (Figure 9A), and slightly reduces body weights of the treated mice compared to those of the vehicle control mice (Figure 9B).

EXAMPLE 12 COMPOUND D SUPRRESSES SURVIVIN EXPRESSION The effect of Compound D on survivin expression was studied at both transcriptional and translational levels. The methods used at the transcriptional level include cDNA microarray analysis, RT-PCR, survivin reporter assays and chromotin immunoprecipitation (ChIP). The methods used at translational levels include Western blot analysis and immunochemistry.

A plasmid containing luciferase under the control of survivin promoter was constructed and transfected into wild type, CBP+/-, or p300+/- 3T3 cells. The results (Figure 10) show that Wnt 1 stimulates expression of the survivin gene in all three types of cells, whereas Compound D reduces expression of the survivin gene and decreases the stimulation of the survinin gene expression by Wnt1 in those cells. Similarly, Compound D and its analog (Comound A) were shown to inhibit expression of survivin in SW480 cells (Figure 11).

Real time reverse transcription-PCR analysis was performed according to the protocol provided with the SYBR Green PCR Master Mix Kit (Perkin Elmer Biosystems, Shelton, ST). Total RNA templates for the RT-PCR reactions were extracted with the RNeasy Midi Kit (Qiagen) from cells treated with Compound D (25 uM) or control (0.5% DMSO) 24 hours after treatment.

The primers used for the RT-PCR reactions were 5'- AGCCCTTTCTCAAGGACCAC-3'and 5'-GCACTTTCTTCGCAGTTTCC-3'.

Table 8 shows the results of the analysis. A ratio less than 0.5 indicates a significant decrease of gene expression due to the treatment of Compound D, whereas a ratio greater than 1.5 indicates a significant increase of gene expression. A ratio about 1 indicates no change. As indicated in Table 8 and Figure 12, the expression of the survivin gene is. significantly reduced in the presence of Compound D compared to the control.

Table 8. Gene Expression with and without Compound D Gene Ratio (Treated/DMSO Control) Ubiquitin 0. 98 GADPH 0. 98 HLAC 1. 01 Survivin 0. 30 PCNA 0. 33 Antigen KI-67 0. 45 MIC-1 7. 0 GADD-153 7. 00 ChIP assays on SW 480 cells treated with either Compound D (25 uM) or control (0.5% DMSO) were performed. As shown in Figure 13, the

survivin promoter is occuried by CBP, p-catenin, Tcf4 and acetylated histone in control treated cells. Treatment with Compound D decreases the association of all these proteins with the survivin promoter.

To characterize the effect of Compound D on the survivin expression at the translational level, Western blot analysis of extracts of cells treated with vehicle (0.5% DMSO) alone, 10 uM or 25 uM Compound D, or 5 uM 5-FU was performed using survivin 6E4 monoclonal antibody (Cell Signaling Technolgy). The results (Figure 14A) show that the treatments with Compound D at both concentrations and the treatment with 5-FU reduced the amount of the survivin protein. The treatments with Compound D at both concentrations were more effective in reducing the survivin expression than the treatment with 5-FU, and the treatment with Compound D at the higher concentration (i. e., 25 M) was most effective.

The effect of Compound D on the survivin expression at the translational level was further characterized using immunofluorescence microscopy. In the absence of Compound D, survivin localizes to the mitotic spindle apparatus, consistent with the notion that survivin is involved in chromosomal separation (Figure 14B). This expression pattern was not observed in SW480 cells after the treatment of Compound D as little or no survivin protein was detected (Figure 14C).

EXAMPLE 13 EFFECTS OF VARIOUS COMPOUNDS ON SURVIVIN AND TCF4 EXPRESSION The effects of various compounds having general formula (I) on survivin and TCF4 expression were characterized. The results are shown in Table 9.

Table 9. Effects of compounds on survivin and TCF4 expression SU V ; V ; n °/O ; nh ; btOn (UM) 5UM 25UM Survivin % inhibition TCF4lC50 (uM) 5uM 25uM i Zozo 100 99 2 OH y- oh OH 1 XOH N-'Y-'--N 97 100-2. 2 ,-I-0 OH OU OH oh / OU N NX 1S N'N\CH 51 93 ~6. 3 NYO N / Survivin % inhibition TCF4 IC50 (hum) 5uM 25uM a N O H3CN'CH3 N" \ 41 92 5. 2 0. 7 0 o OH Nyo CH3 ! H CH3 H3CoN, N, I N CH3 18. 2 2. 4 o o OH N O F F F F H \ 0 80 1. 3 0. 1 OF ° su nOH OH N 0 H3CX N'CH3 Oh H3C,'IN '"'- N 0 93 2. 2 0. 2 0 cri o ci OH Survivin % inhibition TCF4 IC50 _ (uM) 5uM 25uM I a NorO, CH3 N'tN H H3C N N 46 96 4. 4 0. 6 ° su o I v'OH \ (H, CH3 rH r 0,, _, Nyo CH3 9 LJ 77 3. 5_0. 3 T' o I OH \ (H CH3 H I FisCN"w/ . \ f J\CHs l 0 92 7. 3+-0. 6 WOH o ci zu XI v'OH I N 0 H CH Zu N L O uF 79 81 1. 7 0. 2 OF 0 F OH N O H3CCH3 f H ! NNtNwCH3 H3CyN/ N CH3 o 0 84 4. 8 0. 4 0 0 CH3 Ils SOH Survivin % inhibition (uM) (uM) 5uM 25uM I tJ O H3CCH3 H Hz CH3 0 68 10. 9 1. 3 ° spi OH OH N 0 H3CN'CH3 OH H3CNNt\N N 8 4 NA ho Hic OU Nyo H3c, CH3 H zizi 9 91 1. 4 0. 2 -O F 0 'OU zu oh ! H ! CN-tCHs 5 91 6. 3 0. 431 o o OH T OH O H3CN'CH3 N N -OH 0 CH3 OH 'OH Survivin % inhibi, tion TCF4lC50 (ut) 5uM 25uM I aNorO H3CN'CH3 H3CN'N W N Ci 9. ° 0 21 7. 3 _ 1. 1 o OH I 'OH aNof O H3CN'CH3 3 sN | N í ! F ° 91 5. 2_ 1. 1 NYH F H3Cy ( / \ 1 ° S 0 OH Xi 'OU i N-CH3 45 88 13. 2 4. 1 ° su OH I OH aNorO H3CN'CH3 YH Nu H C XH X OH 0 SOH ci /\ \ y cri H CI HsC N N/ 6 58 11. 2 1. 5 o o OH Survivin % inhibition TC (4Ij 50 5uM 25uM CH3 CH3 ° CH CH, CH3 1 48 96 3. 9+-0. 55 0 0 I OH N 0 H3CuNN\NJ H'CI H3CNN \N 9 0 32 50. 4 + 7. 0 - y OH OU cri N 0 w H HsCNNN. 86 91 2. 6 0. 6 ZON I-r 0 OH CH3 N0 CH3 YH HN 9 27 98 10. 7_ 1. 7 LL.''J""' -' ° r OH OH SUrV ; V ; n % ; nh ; b ; t ; On (UM) 5UM 25UM Survivin % inhibition TCF4 IC50 (uM) 5uM 25uM neo ! H/ H3CN' \ 80 97 4. 6 0. 7 lu 0 oh OH XNcO H3C, N-N ° S "OH kJLO"N N'XN 82 97 2. 8 0. 4 0 0 O OH (O H3CNCH3 I H H WN<o uCI 6 89 13. 9 2. 3 Cri 0 O OH hic 0 CH3 f Hz H3C, N'-CH H3C\N ; XCH 14 99 10. 7 1. 9 O 0 OH Survivin % inhibition (uM) (uM) 5uM 25uM H3C,, CH3 I H3 c,'Nii cri H Cl H 3 N tN 25 44 27. 1 4. 6 O 0 1 OH

EXAMPLE 14 COMPOUND D PROMOTES APOPTOSIS VIA SUPPRESSION OF SURVIVIN EXPRESSION To determine the effect of Compound D on apoptosis and the role of survivin in such an effect, the activities of caspases 2 and 3 in cultured tumor cells treated with either Compound D or control were measured. The results (Figure 15) show that (1) Compound D (at 2. 5 uM or 5. 0 UM) activated the caspase 3 activity, but not the caspase 2 activity; (2) stausporine (0.5 uM) increased both the caspase 2 and caspase 3 activities; (3) the co-treatment of stausporine and Compound D produced a synergic stimuation of the caspase 3 activity, but not a synergic stimuation of the caspase 2 activity; and (4) transfection of the survivin gene decreased the activiation of the caspase 3 activity induced by the treatment of stausporine or Compound D, and the synergic stimulation of the caspase 3 activity induced by the co-treatment of stausporine and Compound D. The above results suggest that Compound D stimulate the caspase 3 activity via suppression of the expression of the survivin gene.

The effect of compound D on apoptosis and the role of survivin in such an effect were further characterized by measuring cell death of cultured tumor cells treated with staurosporine (0. 5 uM), Compound D (5. 0 uM) or both.

The results (Figure 16) showed that both Compound D and stausporine promote cell death, and that transfection of the survivin gene decreased the increase in cell death induced by the treatment of stausporine, Compound D, or both. The above results suggest that Compound D promote apoptosis via suppression of the expression of the survivin gene.

To determine the effect of Compound D on cell cycle and the role of survivin in such an effect, FACS analysis was performed on cultured tumor cells with or without transfection of a construct containing the survivin gene and further treated with stausporine (0.5 luM), Compound D (5, uM), or both. The results (Figure 17) show that both stausporine and Compound D increase the number of cells in Go, and that overexpression of survivin in the cells decreases the effect of the treatment of stausporine, Compound D, or both. These results suggest that the effect of Compound D on cell cycle may be at least partially via suppression of the expression of the survivin gene.

It will be appreciated that, although specific embodiments of the invention have been described herein for the purposes of illustration, various modifications may be made without departing from the spirit and scope of the invention. Accordingly, the invention is not limited except by the appended claims.

All of the above U. S. patents, U. S. patent application publications, U. S. patent applications, foreign patents, foreign patent applications and non- patent publications referred to in this specification and/or listed in the Application Data Sheet, including U. S. patent application serial no. 10/087,443 filed on March 01,2002, and U. S. patent application serial no. 09/976,470 filed on October 12,2001, are incorporated herein by reference.

From the foregoing it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims.