Method of drug design

Abstract

The invention provides a method of identifying biologically active compounds comprising: (a) designing a first library of compounds of formula (1) to scan molecular diversity wherein each compound of the library has at least two pharmacophoric groups R1 to R5 as defined below and wherein compound of the library has same number of pharmacophoric groups; (b) assaying the first library of compounds in one or more biological assay(s); and (c) designing a second library wherein each compound of the second library contains one or more additional pharmacophoric group with respect to the first library; such that the/each component of the first and second library is a compound of formula (1).

Claims

1. A method of identifying biologically active compounds comprising: (a) designing a first library of compounds of formula 1 to scan molecular diversity wherein for each compound of the first library R.sub.2 and one of R.sub.1, R.sub.3, R.sub.4, and R.sub.5 as defined below are pharmacophoric groups and the remainder of R.sub.1, R.sub.3, R.sub.4, and R.sub.5 are non-pharmacophoric groups; (b) assessing the first library of compounds in one or more biological activity assay(s); and (c) designing a second library wherein for each compound of the second library the substituents R.sub.1 to R.sub.5 comprise the two pharmacophoric groups identified in step (b) and a third pharmacophoric group, and wherein the remainder of R.sub.1, R.sub.3, R.sub.4, and R.sub.5 are non-pharmacophoric groups, such that the/each component of the first and second library is a compound of formula 1: ##STR00021## wherein Z is sulphur, oxygen, NH, NR.sup.A or hydrogen, in the case where Z is hydrogen then R.sub.1 is not present; R.sup.A is selected from the set defined for R.sub.1, R.sub.3, R.sub.4, and R.sub.5, or wherein Z and R.sub.1 together form a heterocycle; X is oxygen or nitrogen; R.sub.2 is a pharmacophoric group independently selected from the group consisting of C.sub.2 to C.sub.20 alkyl, C.sub.2 to C.sub.20 acyl, wherein said acyl is not acetyl; C.sub.2 to C.sub.20 alkenyl, C.sub.2 to C.sub.20 alkynyl, C.sub.2 to C.sub.20 heteroalkyl, C.sub.5 to C.sub.20 aryl, C.sub.5 to C.sub.20 heteroaryl, C.sub.5 to C.sub.20 arylalkyl and C.sub.5 to C.sub.20 heteroarylalkyl, which is optionally substituted, and can be branched or linear, or wherein NH and R.sub.2 combine to form a heterocycle; R.sub.1, R.sub.3, R.sub.4, and R.sub.5 are each independently selected from (i) non-pharmacophoric groups consisting of H, methyl and acetyl, and (ii) pharmacophoric groups independently selected from the group consisting of C.sub.2 to C.sub.20 alkyl, C.sub.2 to C.sub.20 acyl, wherein said acyl is not acetyl; C.sub.2 to C.sub.20 alkenyl, C.sub.2 to C.sub.20 alkynyl, C.sub.2 to C.sub.20 heteroalkyl; C.sub.5 to C.sub.20 aryl, C.sub.5 to C.sub.20 heteroaryl, C.sub.5 to C.sub.20 arylalkyl and C.sub.5 to C.sub.20 heteroarylalkyl, which is optionally substituted, and can be branched or linear, or wherein X and the corresponding R moiety, R.sub.3 to R.sub.5 respectively, combine to form a heterocycle, and wherein the optional substituent of each of R.sub.2 and, when present as a pharmacophoric group R.sub.1, R.sub.3, R.sub.4, and R.sub.5, are selected from the group consisting of OH, NO, NO.sub.2, NH.sub.2, N.sub.3, halogen, CF.sub.3, CHF.sub.2, CH.sub.2F, nitrile, alkoxy, aryloxy, amidine, guanidiniums, carboxylic acid, carboxylic acid ester, aryl, cycloalkyl, heteroalkyl, heteroaryl, aminoalkyl, aminodialkyl, aminotrialkyl, aminoacyl, carbonyl, imine, sulfate, sulfonamide, phosphate, phosphoramide, hydrazide, hydroxamate, hydroxamic acid, heteroaryloxy, aminoaryl, aminoheteroaryl, thioalkyl, thioaryl and thioheteroaryl.

2. The method according to claim 1, wherein Z is sulphur or oxygen.

3. The method according to claim 1, wherein at least one of the pharmacophoric groups is selected from the group consisting of aryl, arylalkyl, heteroaryl, heteroarylalkyl and acyl.

4. The method according to claim 1, wherein the/each component of each of the libraries is a compound selected from a compound of formula 2 or formula 3 or formula 4: ##STR00022##

5. The method according to claim 1, wherein the/each compound of the first library is of the gluco- or galacto-configuration.

6. The method according to claim 4, wherein the/each compound of the second library is of the gluco- or galacto- or allo-configuration.

7. The method according to claim 6, wherein the/each compound of the second library is of the gluco-configuration.

8. The method according to claim 6, wherein the/each compound of the second library is of the allo-configuration.

9. The method according to claim 6, wherein the/each compound of the second library is of the galacto-configuration.

10. The method according to claim 1, wherein the biological assays involve peptide ligand class of GPCRs.

11. The method according to claim 1, wherein NH and R.sub.2 combine to form a heterocycle.

12. The method according to claim 11, wherein the heterocycle is heteroaryl.

13. The method according to claim 11, wherein the heterocycle is selected from group consisting of triazoles, benzimidazoles, benzimidazolone, benzimidazolothione, imidazole, hydantoine, thiohydantoine and purine.

14. The method according to claim 1, wherein in the first library one pharmacophoric group comprises a positive charge and one pharmacophoric group is selected from the group consisting of aryl, heteroaryl, arylalkyl and heteroarylalkyl.

15. The method according to claim 1, wherein the method further comprises the steps of (d) assaying the second library of compounds in one or more biological assay(s); and (e) designing a third library wherein for each compound of the third library the substituents R.sub.1 to R.sub.5 comprise the three active pharmacophoric groups identified in step (c) and a fourth pharmacophoric group, and wherein each component of the third library is a compound of formula 1.

16. The method according to claim 1, wherein in each library when X is oxygen, R.sub.1, R.sub.3, R.sub.4 and R.sub.5 are each independently selected from (i) non-pharmacophoric groups consisting of H, and methyl, and (ii) pharmacophoric groups independently selected from the group consisting of C.sub.2 to C.sub.20 alkyl, C.sub.2 to C.sub.20 alkenyl, C.sub.2 to C.sub.20 alkynyl, C.sub.2 to C.sub.20 heteroalkyl, C.sub.5 to C.sub.20 aryl, C.sub.5 to C.sub.20 heteroaryl, C.sub.5 to C.sub.20 arylalkyl, and C.sub.5 to C.sub.20 heteroarylalkyl, which is optionally substituted, and can be branched or linear; and, when X is nitrogen, R.sub.1, R.sub.3, R.sub.4 and R.sub.5 are each independently selected from (i) a non-pharmacophoric group which is acetyl, and (ii) pharmacophoric groups independently selected from the group consisting of C.sub.2 to C.sub.20 alkyl, C.sub.2 to C.sub.20 acyl wherein said acyl is not acetyl; C.sub.2 to C.sub.20 alkenyl, C.sub.2 to C.sub.20 alkynyl, C.sub.2 to C.sub.20 heteroalkyl, C.sub.5 to C.sub.20 aryl, C.sub.5 to C.sub.20 heteroaryl, C.sub.5 to C.sub.20 arylalkyl, and C.sub.5 to C.sub.20 heteroarylalkyl, which is optionally substituted, and can be branched or linear.

17. The method according to claim 16, wherein Z is sulphur or oxygen.

18. The method according to claim 16, wherein the/each component of each of the libraries is a compound selected from a compound of formula 2 or formula 3 or formula 4: ##STR00023##

19. The method according to claim 17, wherein the/each compound of the second library is of the gluco- or galacto- or allo-configuration.

20. The method according to claim 16, wherein the/each compound of the first library is of the gluco- or galacto-configuration.

21. The method according to claim 16, wherein in the first library one pharmacophoric group comprises a positive charge and one pharmacophoric group is selected from the group consisting of aryl, heteroaryl, arylalkyl and heteroarylalkyl.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) The embodiments of the invention will be described with reference to the following examples. Where appropriate, the following abbreviations are used.

(2) Ac Acetyl

(3) DTPM 5-Acyl-1,3-dimethylbarbiturate

(4) Ph Phenyl

(5) TBDMS t-Butyldimethylsilyl

(6) TBDPS t-Butyldiphenylsilyl

(7) Bn benzyl

(8) Bz benzoyl

(9) Me methyl

(10) DCE 1,2-dichloroethane

(11) DCM dichloromethane, methylene chloride

(12) Tf trifluoromethanesulfonyl

(13) Ts 4-methylphenylsulfonyl, p-toluenesulfonyl

(14) DMF N,N-dimethylformamide

(15) DMAP N,N-dimethylaminopyridine

(16) ,-DMT ,-dimethoxytoluene, benzaldehyde dimethyl acetal

(17) DMSO dimethylsulfoxide

(18) DTT dithiothreitol

(19) DMTST Dimethyl(methylthio)sulphoniumtrifluoro-methanesulphonate

(20) TBAF tetra-n-butylammonium fluoride

Part A

Preparation of Building Blocks

(21) In order to fully enable the invention, there is described below methods for the preparation of certain building blocks used in the preparation of the compounds of the invention. The building blocks described are suitable for both solution and solid phase synthesis of the compounds of the invention.

Example A: Synthesis of a 2,4 Dinitrogen Containing Galactopyranoside Building Block

(22) ##STR00003##

Example B: Synthesis of a 3-nitrogen Containing Gulopyranoside Building Block

(23) ##STR00004##

Example C: Synthesis of a 2,6-dinitrogen Substituted Glucopyranoside Building Block

(24) ##STR00005##

Example D: Synthesis of a 2-nitrogen Containing Tallopyranoside Building Block

(25) ##STR00006##

Example E: Synthesis to 3-nitrogen Containing Altropyranoside Building Block

(26) ##STR00007##

Example F: Synthesis 2-nitrogen Containing Glucopyranoside Building Block

(27) ##STR00008##

(28) ##STR00009##

Example G: Synthesis of a 2-nitrogen Containing Allopyranoside Building Block

(29) ##STR00010##

(30) The Solid Phase Library Synthesis of Sugars is illustrated in Scheme 1.

(31) The reaction conditions are as follows:

(32) (A) 2P Compound Synthesis: R.sub.1=R.sub.2=OMe;

(33) i) 2-naphthalene methanol, DMTST, DCM; TCA-Wang resin, BF.sub.3.Et.sub.2O, DCM; iii) NaOMe, methanol; iv a. KOtBu, DMF; b. Mel, DMF; v) HF proton sponge, AcOH, DMF, 65 C.; vi) a. KOtBu, DMF; b. Mel, DMF; vii) 1,4-dithio-DL-threitol, KOtBu, DMF; viii) HBTU, Fmoc--Ala-OH, DIPEA, DMF; ix) piperidine/DMF (); x) TFA, Et.sub.3SiH, DCM
(B) 3P Compound Synthesis: R.sub.1=methyl-2-naphthyl, R.sub.2=OMe;
i) 2-naphthalene methanol, DMTST, DCM; ii) TCA-Wang resin, BF.sub.3.Et.sub.2O, DCM; iii) NaOMe, methanol; iv) a. KOtBu, DMF; b, 2-bromomethyl-naphthalene, DMF; v) HF proton sponge, AcOH, DMF, 65 C.; vi) a. KOtBu, DMF; b. Mel, DMF; vii) 1,4-dithio-DL-threitol, KOtBu, DMF; viii) HBTU, Fmoc--Ala-OH, DIPEA, DMF; ix) piperidine/DMF (), x) TFA, Et.sub.3SiH, DCM
(C) 4P Compound Synthesis: R.sub.1=methyl-2-naphthyl, R.sub.2=4-chlorobenzyl
i) 2-napthalene methanol, DMTST, DCM; ii) TCA-Wang BF.sub.3.Et.sub.2O, DCM; iii) NaOMe, methanol; iv) a. KOtBu, DMF; b. 2-bromomethyl-naphthalene, DMF; v) HE proton sponge, AcOH, DMF, 65 C.; vi) a. KOtBu, DMF; b. 4-chlorobenzylbromide, DMF; vii) 1,4-dithio-DL-threitol, KOtBu, DMF; viii) HBTU, Fmoc--Ala-OH, DIPEA, DMF; ix) piperidine/DMF (); x) TFA, Et.sub.3SiH, DCM

(34) ##STR00011## ##STR00012##

(35) ##STR00013##

(36) The synthesis of the Allose 2,6N building block is illustrated in Scheme 2. The reaction conditions are as follows:

(37) i) p-methoxybenzaldehyde dimethylacetal, camphorsulfonic acid, N,N-dimethylformamide (DMF); ii) Tf.sub.2O, pyridine, dichloromethane (DCM); iii) tetrabutylammonium benzoate, DMF, 55 C.; iv) BH.sub.3.THF, Bu.sub.2BOTf, DCM; v) methanesulfonylchloride, pyridine, DCM; vi) sodium azide, DMF, 85 C.; vii) sodium methanolate (NaOMe), methanol; viii) n-butanol, ethylene diamine, reflux; ix) DTPM reagent, methanol; x) benzoic anhydride, pyridine xi) trifluoroacetic acid, triethylsilane, DCM
Designing Libraries

(38) The design of the libraries is based on the presentation of a positive charge and a crying number of aromatic substituents in different spatial arrangements on a monosaccharide scaffold. Starting with a positive charge and one aromatic displayed on the core scaffold, actives from this first library were elaborated on by further variation and addition of more aromatic substituents to quickly identify highly active molecules.

(39) The first library of compounds comprises two pharmacophoric groups, known as a 2P library, in particular, one containing an aromatic and a positive charge. The library was designed such that each molecule presents two pharmacophoric groups in different relative orientation or presentation (e.g., distance, relative angle, i.e. relative position in, space is different).

(40) Actives from this library were identified and SAR information from this library was used to design subsequent library of compounds wherein each compound may include three pharmacophoric groups, known as a 3P library. Subsequent libraries with four pharmacophoric groups are called 4P library, etc.

(41) Members of significantly improved activity were identified out of the second library and were selected for further drug development.

(42) The method of the invention includes real and virtual libraries.

(43) Thus, the molecules according to formula 1 are well suited for generating iterative scanning libraries, starting from a selected number of pharmacophores (eg, two) in the first library and designing subsequent libraries with additional pharmacophores based on SAR information from the first library, thereby assisting in delineating pharmacophores.

(44) The 2P and 3P library of compounds were synthesized according to the budding blocks as described in Examples A-G.

(45) The 2P library (Table 1) was designed to scan molecular diversity for 3P molecules, comprising an aromatic and a positive charge.

(46) The 2P library was screened for biological activity and the results are given in Table 1.

(47) Similarly, the 3P library was designed to scan molecular diversity for 3P molecules. Design of 3P library resulted from SAR obtained from 2P library in Table 1.

(48) The 3P library was screened for biological activity and the results are given in Table 2.

(49) A visual analysis of the results according to Table 1 (2P library) and Table 2 (3P Library) indicates that:

(50) 1. 1, 2 allose substitution according to formula 0.3 (and Scaffold C/D) presents the most active arrangement of molecules in the library wherein Z is oxygen, R.sub.1 is naphthyl and R.sub.2 is propylamine or ethylamine.

(51) These compounds represent most actives at low mM range, and are suitable candidates for further drug development.

(52) 2. R.sub.1 as naphthyl is more active than the corresponding p-chlorobenzyl substituent.

(53) 3. 1, 2 allose according to formula 3 (Scaffold C/D) is more active than the corresponding 1, 2 glucose conformation (Scaffold A/B).

(54) 4. 1. 2 substitution according to formula 3 (Scaffold C/D) is more active then the corresponding 2, 6 substitution according the formula 4 (Scaffold G)

(55) 5. R.sub.2 as propylamine and ethylamine are more active than methylamine wherein Z, R.sub.1 and R.sub.2 are as described above.

(56) 6. 2, 3 allose substitution according to formula 3 (Scaffold C/D) presents the more actives wherein R.sub.2 is ethylamine, and R.sub.3 is p-chlorobenzyl compared to corresponding R.sub.2 as propylamine and ethylamine wherein R3 is p-chlorobenzyl substituent, and also wherein R.sub.2 is methylamine, ethylamine or propylamine and R3 is naphthyl.
7. 2, 3 glucose substitution according to formula 3 (scaffold A/B) presents the more actives wherein R.sub.2 propylamine and R.sub.3 is naphthyl compared to corresponding R.sub.2 as methylamine or ethylamine, and also wherein R.sub.2 is methylamine, ethylamine or propylamine and R.sub.3 is p-chlorobenzyl.
8. 2, 4 and 3, 4 substitutions according to formula 3 (Scaffold G) present the least actives.

Part B

Biological Assays

Example H: In Vitro Screening of Compounds Against Somatostatin Subtypes SSTR-1 to SSTR-5

(57) General Method

(58) Receptor membrane preparations containing the desired cloned receptor (for example cloned human somatostatin receptor subtype 5, SSTR5) and radiolabeled ligand were diluted at the concentration required for testing and according to the specific parameters associated with the selected receptor-ligand combination, including receptor B.sub.max, ligand K.sub.d and any other parameters necessary to optimize the experimental conditions. When tested for competition activity to the reference ligand, compound was mixed with membrane suspension and the radiolabeled reference ligand (with or without an excess of unlabeled ligand to the receptor for determination of non-specific binding) and incubated at the temperature required by internal standard operating procedures. Following incubation, the binding reaction was stopped by the addition of ice-cold washing buffer and filtered on appropriate filters, which are then counted. Data analysis and curve-fitting was performed with XLfit (IDBS).

(59) Preparation of Compounds

(60) 10 mM solutions of test compounds in 100% DMSO were prepared, 160 l was used for each dilution (20 l/well in triplicate).

(61) A 1.25 mM assay stock was prepared by making a 1:8 dilution of the 10 mM solution. To 30 L of the 10 mM solution was added 210 L milli-Q H.sub.2O. A 1:5 dilution series in milli-Q H.sub.2O was then prepared.

(62) TABLE-US-00001 Final concentration Final concentration in SST4 assay in SST5 assay A. 240 L of 1.25 mM 0.25 mM 0.125 mM B. 48 L A + 192 L mQ 0.05 mM 0.025 mM C. 24 L B + 192 L mQ 0.01 mM 0.005 mM etc

(63) Assays were performed in triplicate at each concentration within the 1:5 dilution series: 250 M, 50 M, 10 M, 2 mM, 0.4 M, 0.08 M, 0.016 M, 0.0032 M, etc. (for SST4 assay and 125 M, 10 M, 2 M, 1 M, 0.5 M, etc (for SST5 assay).

(64) Fitter Plate Assay for SST5 Receptor

(65) Human SST5 somatostatin receptor was transfected into HEK-293 EBNA cells. Membranes were suspended in assay buffer (50 mM Tris-HCl, 1 mM EGTA, 5 mM MgCl.sub.2, 10% sucrose, pH 7.5). The receptor concentration (B.sub.max) was 0.57 pmol/mg protein K.sub.d for [.sup.125I]SST-14 Binding 0.31 nM, volume 0.4 ml per vial (400 microassays/vial), and protein concentration 1.03 mg/ml.

(66) After thawing the frozen receptor preparation rapidly, receptors were diluted with binding buffer, homogenized, and kept on ice. 1. Use Multiscreen glass fiber filter plates (Millipore, Cat No MAFCNOB10) precoated with 0.5% PEI for 2 hr at 4 C. Before use add 200 l/well assay buffer and filter using Multiscreen Separation System. 2. Incubate 5.5 g of membranes (40 l of a 1:40 dilution), buffer and [.sup.125I]SST-14 (4 nM, 80 000 cpm, 2000 Ci/mmol) in a total volume of 200 l for 60 min at 25 C. Calculate IC50 for SST-14 (a truncated version of the natural ligand SST-28) (Auspep, Cat No 2076) and SST-28 (Auspep, Cat No 1638). Prepare serial dilutions (1:5) of compounds, as described above and instead of adding SST-14 in well, add 20 l of compounds (Table 3). 3. Filter using Multiscreen Separation System with 50.2 ml ice-cold Assay buffer. 4. Remove the plastic underdrain and dry plate in oven for 1 hr at 40 C. 5. Seal tape to the bottom of the plate. 6. Add 50 l/well scintillant (Supermix, Wallac, Cat No 1200-439). 7. Seal and count in the BJET, program 2.

(67) TABLE-US-00002 TABLE 3 Compounds Volume (ul) TB NSB testing Membranes (5.5 g/well) 40 40 40 Radio-labeled label (~80 000 40 40 40 cpm, ~4 nM) Unlabeled ligand 20 mQH.sub.2O 20 Compounds 20 Assay buffer 100 100 100 Total volume (I) 200 200 200 TB: total binding NSB: non-specific binding

Part C

General Experimental Methods

Example I: HPLC Method for Compounds in Tables 1 and 2

(68) The HPLC separation of compounds in Tables 1 and 2 was conducted under Method A or Method B as shown below.

(69) Method A

(70) Column: Agent SB Zorbax C18 4.650 mm (5 m, 80 )

(71) LC mobile phase:

(72) 5% aqueous MeCN/1 min

(73) 100% MeCN/7-12 min

(74) Method B

(75) Column: Agilent SB Zorbax C18 4.650 mm (5 m, 80 )

(76) LC mobile phase:

(77) 5% aq MeCN/1 min

(78) 30% aq MeCN/3 min

(79) 40% aq MeCN/12 min

(80) 100% MeCN/13-15 min

(81) Key to Building Blocks for Tables 1 and 2

(82) Table 1: *% SST5 radio-ligand binding displaced at conc (M) for 2P library of compounds

(83) Table 2: *% SST5 radio-ligand binding displaced at conc (M) for 3P library of compounds; R.sub.4=X30; compounds 60-63, 119 and 156-159 are comparative compounds from 2P library

(84) ++: % SST5 radio-ligand binding displaced at conc (M) >60%

(85) +: % SST5 radio-ligand binding displaced at conc (M) 60>+>40%

(86) : % SST5 radio-ligand binding displaced at conc (M) <40%

(87) Blank: not determined

(88) RT: retention time/minutes

(89) M+H: mass ion+1

(90) TABLE-US-00003 TABLE 1 Biological activity of example 2P library conc conc conc Object ID Scaffold R1 R2 R3 R4 R5 500 250 50* RT M + H 1 E X15 X2 X30 X24 3.24 449.58 2 A X7 X20 X24 X30 X24 3.4 383.46 3 A X7 X15 X24 X30 X24 3.42 397.48 4 E X20 X2 X30 X24 3.49 435.55 5 A X2 X20 X24 X30 X24 ++ + 3.88 419.21 6 A X2 X15 X24 X30 X24 ++ + 3.83 433.23 7 E X19 X24 X30 X3 3.42 405.12 8 E X19 X24 X30 X2 ++ + 3.81 421.17 9 E X19 X3 X30 X24 3.62 405.12 10 E X19 X2 X30 X24 4.03 421.17 11 A X3 X19 X24 X30 X24 3.39 389.14 12 A X2 X19 X24 X30 X24 4.08 405.19 13 B X3 X19 X24 X30 X24 3.4 389.14 14 B X2 X19 X24 X30 X24 3.88 405.19 15 E X20 X24 X30 X3 3.25 419.13 16 E X20 X24 X30 X2 + 3.59 435.19 17 E X20 X3 X30 X24 3.68 419.13 18 E X20 X2 X30 X24 4.06 435.19 19 A X3 X20 X24 X30 X24 ++ 3.56 403.16 20 B X3 X20 X24 X30 X24 + 3.37 403.16 21 B X2 X20 X24 X30 X24 ++ + 3.7 419.21 22 E X15 X24 X30 X3 3.22 433.15 23 E X15 X24 X30 X2 + 3.59 449.2 24 E X15 X3 X30 X24 3.7 433.15 25 E X15 X2 X30 X24 + 4.06 449.2 26 E X3 X15 X24 X30 X24 ++ 3.57 417.17 27 B X3 X15 X24 X30 X24 3.4 417.17 28 B X2 X15 X24 X30 X24 ++ 3.68 433.23 29 F X19 X24 X30 X3 3.55 405.12 30 F X19 X24 X30 X2 + 3.84 421.17 31 F X19 X3 X30 X24 + 3.75 405.12 32 F X19 X2 X30 X24 4.05 421.17 33 C X3 X19 X24 X30 X24 3.38 389.14 34 C X2 X19 X24 X30 X24 3.72 405.19 35 D X3 X19 X24 X30 X24 3.41 389.14 36 D X2 X19 X24 X30 X24 + 3.77 405.19 37 F X20 X3 X30 X24 3.76 419.13 38 C X3 X20 X24 X30 X24 ++ + 3.33 403.16 39 D X3 X20 X24 X30 X24 ++ 3.44 403.16 40 D X2 X20 X24 X30 X24 ++ ++ 3.8 419.21 41 F X15 X24 X30 X3 3.51 433.15 42 F X15 X24 X30 X2 + 3.81 449.2 43 F X15 X3 X30 X24 3.66 433.15 44 D X3 X15 X24 X30 X24 ++ 3.51 417.17 45 D X2 X15 X24 X30 X24 ++ + 3.86 433.23 46 G X24 X3 X19 X30 3.31 386.14 47 G X19 X2 X24 X30 3.27 402.2 48 G X19 X24 X8 X30 2.48 352.18 49 G X2 X24 X19 X30 3.64 388.18 50 G X8 X24 X19 X30 2.61 352.18 51 G X24 X3 X20 X30 3.08 400.16 52 G X2 X24 X20 X30 3.46 402.2 53 G X8 X24 X20 X30 2.73 366.2 54 G X24 X3 X15 X30 3.27 414.17 55 G X2 X24 X15 X30 3.79 416.21 56 G X8 X24 X15 X30 2.78 380.21 57 F X20 X2 X30 X24 4.01 435.19 58 F X15 X2 X30 X24 4.08 449.2 59 C X2 X20 X24 X30 X24 ++ ++ + 3.74 419.21

(91) TABLE-US-00004 TABLE 2 Biological activity of example 3P library Object conc conc conc conc conc conc conc conc conc ID Scaffold R1 R2 R3 R5 500 250 50 10 1.0 0.5 0.25 0.10 0.001* RT M + H 60 A X2 X20 X24 X24 ++ + 3.88 419.21 61 B X2 X20 X24 X24 ++ + 3.7 419.21 62 D X2 X20 X24 X24 ++ ++ 3.8 419.21 63 C X2 X20 X24 X24 ++ ++ + 3.72 419.21 64 C and D X2 X20 X8 X24 ++ ++ ++ + 4.98 65 C and D X2 X20 X8 X24 ++ 4.98 66 C and D X2 X20 X3 X24 ++ ++ ++ 5.25 67 C and D X2 X20 X3 X24 ++ 5.25 68 C and D X2 X20 X1 X24 ++ ++ ++ 5.49 69 C and D X2 X20 X2 X24 ++ ++ ++ ++ + 5.23 70 C and D X2 X20 X3 X2 + 5.85 71 C and D X2 X20 X3 X8 ++ 5.61 72 C and D X2 X20 X3 X3 ++ 5.51 73 C and D X2 X20 X2 X2 + 5.95 74 C and D X2 X20 X2 X8 ++ 5.45 75 C and D X2 X20 X2 X3 ++ 6.46 76 C and D X2 X20 X8 X2 ++ 5.7 77 C and D X2 X20 X8 X8 ++ 5.01 78 C and D X2 X20 X8 X3 ++ + 5.37 79 B X2 X20 X2 X2 ++ 10.31 80 A X2 X20 X2 X2 ++ 10.88 81 B X2 X20 X2 X8 ++ 8.02 82 A X2 X20 X2 X8 ++ + 8.68 83 B X2 X20 X2 X3 ++ 9.39 84 A X2 X20 X2 X3 ++ 10.24 85 D X2 X20 X2 X24 ++ ++ 50.92 86 C X2 X20 X2 X24 ++ ++ 54.37 87 A or B X2 X20 X8 X24 3.78 495.59 88 A or B X2 X20 X8 X24 3.86 495.59 89 A or B X2 X20 X3 X24 3.95 530.03 90 A or B X2 X20 X3 X24 ++ + 3.97 530.03 91 A or B X2 X20 X1 X24 4.5 571.69 92 A or B X2 X20 X2 X24 + 4.33 545.65 93 A and B X2 X20 X24 X8 + 4.13 495.59 94 A or B X2 X20 X24 X3 4.33 530.03 95 A or B X2 X20 X24 X3 4.33 530.03 96 A or B X2 X20 X24 X1 4.77 571.69 97 A and B X2 X20 X24 X2 + 4.52 545.65 98 A X2 X20 X2 X24 ++ + 5.45 545.65 99 A X2 X31 X2 X24 + 5.07 559.67 100 A X2 X32 X2 X24 ++ + 5.05 559.67 101 A X2 X33 X2 X24 + 4.79 557.66 102 A X2 X34 X2 X24 6.24 613.77 103 A X2 X35 X2 X24 ++ + 5.85 585.71 104 A X2 X36 X2 X24 6.33 599.74 105 A X2 X37 X2 X24 6.72 599.74 106 A X2 X45 X2 X24 4.96 573.7 107 A X2 X20 X46 X24 ++ ++ 4.22 530.03 108 A X2 X20 X47 X24 ++ + 4.87 564.48 109 A X2 X20 X48 X24 ++ 4.98 530.03 110 A X2 X20 X49 X24 ++ ++ 4.43 546.64 111 A X2 X20 X50 X24 5.44 552.66 112 A X2 X20 X51 X24 ++ + 3.78 546.64 113 A X2 X20 X52 X24 ++ ++ 5.71 564.48 114 A X2 X20 X9 X24 ++ ++ 5.89 545.65 115 A X2 X20 X53 X24 ++ + 5.8 564.48 116 A X2 X20 X54 X24 ++ + 4.43 546.64 117 A X2 X20 X55 X24 ++ ++ 5.71 564.48 118 A X2 X20 X56 X24 ++ ++ 6.9 587.68 119 A X2 X15 X24 X24 ++ + 120 A and B X2 X15 X8 X24 ++ + 4.29/4.57 121 A and B X2 X15 X24 X1 + + 5.4 122 A and B X2 X15 X24 X2 ++ ++ 5.18 123 A and B X2 X15 X24 X8 4.78 124 A and B X2 X15 X24 X3 + 5.07 125 A and B X2 X15 X24 X4 + 4.28 126 C and D X2 X15 X8 X24 ++ + + 4.97 127 C and D X2 X15 X3 X24 ++ ++ ++ 5.17 128 C and D X2 X15 X1 X24 ++ + ++ 5.45 585.71 129 C and D X2 X15 X2 X24 ++ ++ + 5.18 559.67 130 A and B X2 X15 X4 X24 ++ 131 A and B X2 X15 X1 X24 ++ 132 A and B X2 X15 X2 X24 ++ 133 A and B X2 X15 X3 X24 ++ 134 A X2 X15 X3 X24 ++ ++ ++ ++ + 4.78 135 A X2 X15 X3 X2 ++ 9.87 136 A X2 X15 X3 X8 ++ 7.82 137 A X2 X15 X3 X3 ++ 9.32 138 A X2 X38 X2 X24 3.67 574.69 139 A X2 X39 X2 X24 + 5.07 573.7 140 A X2 X40 X2 X24 ++ ++ 4.96 573.7 141 A X2 X41 X2 X24 5.16 587.73 142 A X2 X53 X2 X24 ++ + 5.69/7.43 599.74 143 A X2 X42 X2 X24 5.98 613.77 144 A X2 X15 X46 X24 ++ + 4.34 544.06 145 A X2 X15 X47 X24 ++ + 5.07 578.5 146 A X2 X15 X48 X24 ++ 5.05 544.06 147 A X2 X15 X49 X24 ++ + 4.5 560.66 148 A X2 X15 X50 X24 5.34 566.69 149 A X2 X15 X51 X24 + 3.95 560.86 150 A X2 X15 X52 X24 ++ ++ 5.78 578.5 151 A X2 X15 X9 X24 ++ + 5.78 559.67 152 A X2 X15 X53 X24 ++ + 5.97 578.5 153 A X2 X15 X54 X24 ++ ++ 4.32 580.66 154 A X2 X15 X55 X24 ++ ++ 5.88 578.5 155 A X2 X15 X56 X24 ++ ++ 7.25 601.71 156 A X3 X19 X24 X24 3.39 389.14 157 B X3 X19 X24 X24 158 C X3 X19 X24 X24 3.38 389.14 159 D X3 X19 X24 X24 160 C and D X3 X19 X8 X24 4.8 161 C and D X3 X19 X3 X24 ++ 5.14 162 C and D X3 X19 X1 X24 + 5.45 542.04 163 C and D X3 X19 X2 X24 ++ 5.2 164 C and D X3 X43 X24 X2 + 3.45 165 C and D X3 X44 X24 X2 ++ 4 166 A and B X3 X43 X24 X2 ++ 3.59 167 A and B X3 X44 X24 X2 ++ ++ 3.97 168 A or B X3 X19 X8 X24 169 A or B X3 X19 X8 X24 170 A or B X3 X19 X3 X24 171 A or B X3 X19 X3 X24 172 A or B X3 X19 X1 X24 173 A or B X3 X19 X1 X24 174 A or B X3 X19 X2 X24 175 A or B X3 X19 X2 X24 176 A and B X3 X19 X24 X8 4.88/5.61 465.95 177 A and B X3 X19 X24 X3 6.06/6.52 500.39 178 A and B X3 X19 X24 X1 9.09 542.04 179 A and B X3 X19 X24 X2 7.43 516.01
Figure 1: Sidearms for Tables 1 and 2

(92) ##STR00014## ##STR00015## ##STR00016## ##STR00017## ##STR00018## ##STR00019## ##STR00020##

(93) Throughout the specification and the claims (if present), unless the context requires otherwise, the term comprise, or variations such as comprises or comprising, will be understood to apply the inclusion of the stated integer or group of integers but not the exclusion of any other integer or group of integers.

(94) Throughout the specification and claims (if present), unless the context requires otherwise, the term substantially or about will be understood to not be limited to the value for the range qualified by the terms.

(95) It should be appreciated that various other changes and modifications can be made to any embodiment described without departing from the spirit and scope of the invention.

REFERENCES

(96) [1] Patel, Y. C. (1999) Somatostatin and its receptor family. Front. Neuroendocr. 20, 157-198 [2] Csaba, Z. and Dournaud, P. (2001) Cellular biology of somatostatin receptors. Neuropeptides 35, 1-23 [3] T. Reisine, T. (1995) Somatostatin receptors: Am. J. Pysiol. (Gastrointest. Liver Physiol. 32) 269, G813-G820 [4] Bauer, W. et al. (1982) SMS201-995: A very potent and selective octapeptide analogue of somatostatin with prolonged action. Life Sci. 31, 1133-1140 [5] Lamberts, S. W. J. et al. (1996) Drug therapy: Octreotide. N. Eng. J. Med. 334, 246-254 [6] Robinson, C. and Castaner, J. (1994) Lanreotide acetate. Drugs Future 19, 992-999 [7] Reisine, T. and Bell. G. I. (1995) Molecular biology of somatostatin receptors. Endocr. Rev. 16, 427-442