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WO1997001560A1 - Banque combinatoire de 1,4-benzodiazepine-2,5-diones - Google Patents

Banque combinatoire de 1,4-benzodiazepine-2,5-diones Download PDF

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Publication number
WO1997001560A1
WO1997001560A1 PCT/US1996/011070 US9611070W WO9701560A1 WO 1997001560 A1 WO1997001560 A1 WO 1997001560A1 US 9611070 W US9611070 W US 9611070W WO 9701560 A1 WO9701560 A1 WO 9701560A1
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WIPO (PCT)
Prior art keywords
resin
formula
linked
aminoacids
azidobenzoyl
Prior art date
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PCT/US1996/011070
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English (en)
Inventor
Michael H. J. Ohlmeyer
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Pharmacopeia LLC
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Pharmacopeia Inc
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Publication date
Application filed by Pharmacopeia Inc filed Critical Pharmacopeia Inc
Priority to EP96923532A priority Critical patent/EP0850238A4/fr
Priority to AU64020/96A priority patent/AU6402096A/en
Priority to JP9504592A priority patent/JPH11508563A/ja
Publication of WO1997001560A1 publication Critical patent/WO1997001560A1/fr
Priority to US08/996,579 priority patent/US5962337A/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
    • C07D243/16—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals
    • C07D243/18—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals substituted in position 2 by nitrogen, oxygen or sulfur atoms
    • C07D243/24—Oxygen atoms

Definitions

  • the present invention relates to a method of synthesizing a combinatorial library of 1 ,4-benzodiazepin-5-ones on solid supports via an aza-Wittig ring closure, said compounds optionally encoded with tags, and to the use of this library in assays to discover biologically active compounds, and, optionally, to cleave 1,4-benzodiazepin-2,5-diones therefrom.
  • T'-L- is an identifier residue
  • -L'-II' is a linker/ligand residue
  • R 1 is H, lower alkyl, c-lower alkyl, -or (CH 2 ) m R 4 , or R 1 and R 2 , together with the atoms to which they are attached, join to form a 5-, or 6-membered heterocyclic ring, optionally monosubstituted with OH, alkoxy, or arylalkoxy;
  • R 2 is H, loweralkyl, arylR 6 R 7 R 8 , or heteroarylR 6 R 7 R 8 , or R 1 and R 2 , together with the atoms to which they are attached, join to form a 5- or 6-membered heterocyclic ring, optionally monosubstituted with OH, alkoxy, or arylalkoxy;
  • R 3 is H or loweralkyl;
  • R 4 is aryl, substituted aryl, heteroaryl, substituted heteroaryl,
  • R 5 is H, lower alkyl, -CNHR 3 R 3 , or -C(O)R 3 ;
  • R 6 , R 7 , and R 8 is each, independently, H, lower alkyl, lower alkoxy, halogen, aryl, lower alkylthio, X-aryl, X-substituted aryl, lower alkylaryl, C(hal) 3 , -(CH 2 ) m NR 3 R 5 , or
  • X is O or S.
  • Preferred libraries of Formula I are those wherein
  • T'-L- is of the Formula
  • n 3-12;
  • Ar is halophenyl
  • the ligands of Formula II may be detached by photolytic, oxidative, acidic, basic, or other cleavage techniques.
  • acidic cleavage may be represented by:
  • L" is the residue from L' and II'OH is II, in its tautomeric amide form:
  • An embodiment of the invention is the solid phase synthesis of 1,4-benzodiazepin-2,5-diones via aza-Wittig ring closure.
  • the process comprises:
  • step (c) cyclizing the resin linked N-(2-azidobenzoyl)amino esters of step (c) via aza-Wittig ring closure to form resin linked benzodiazepines; and, optionally,
  • step (d) cleaving the resin linked benzodiazepines of step (d) to form 1,4-benzodiazepin-2,5-diones.
  • a preferred embodiment of the invention is the solid phase synthesis of 1,4-benzodiazepin-2,5-diones via aza-Wittig ring closure, wherein the process comprises: a) reacting a set of suitably protected ⁇ -aminoacids of the formula:
  • step (b) reacting the resin linked aminoacids of step (a), suspended in DMF and acetic acid, with a set of aldehydes of the formula HC(O)R 2 in HOAc/DMF and sodium cyanoborohydride in THF to form resin linked N-alkyl- ⁇ -aminoacids of the formula: ;
  • step (c) treating the resin linked N-(2-azidobenzoyl)amino esters of step (c), suspended in an involatile solvent (i.e., a non-protic, organic solvent with a boiling point of 80-140 C) such as toluene, xylene, or chlorobenzene, with an excess of a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine at 80-150°C and then cooling said mixture to room temperature to form resin linked benzodiazepines of formula:
  • an involatile solvent i.e., a non-protic, organic solvent with a boiling point of 80-140 C
  • a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine at 80-150°C
  • step (d) suspending the resin linked benzodiazepines of step (d) in TF A/water at room temperature for 1-24 hours to form 1,4-benzodiazepin-2,5-diones of formula:
  • a second preferred embodiment of the invention is the solid phase synthesis of 1,4-benzodiazepin-2,5-diones and 1,3-cyclo-1,4-benzodiazepin-2,5-diones via aza-Wittig ring closure, wherein the process comprises:
  • step (b) reacting the resin linked N-alkyl- ⁇ -aminoacids of step (b), in methylene chloride and diisopropylethylamine, with 2-azidobenzoyl chlorides of formula: X
  • step (b) treating the resin linked N-(2-azidobenzoyl)amino esters of step (b), suspended in an involatile solvent (i.e., a non-protic, organic solvent with a boiling point of 80-140 C) such as toluene, xylene, or chlorobenzene, with an excess of a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine at 80-150°C and then cooling said mixture to room temperature to form resin linked benzodiazepines of formula:
  • an involatile solvent i.e., a non-protic, organic solvent with a boiling point of 80-140 C
  • a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine at 80-150°C
  • step (c) suspending the resin linked benzodiazepines of step (c) in TF A/water at room temperature for 1-24 hours to form 1,4-benzodiazepin-2,5-diones of formula:
  • an aspect of the invention is a method of identifying a compound having a desired characteristic which comprises synthesizing a combinatorial library of Formula I and testing the library of Formula I, either attached to the solid support or detached therefrom, in an assay which identifies compounds of Formula II having the desired characteristic.
  • Another embodiment of the invention is a method of identifying a compound having a desired characteristic which comprises testing the library of Formula I, either attached to the solid support or detached therefrom, in an assay which identifies compounds of Formula II having the desired characteristic.
  • a further embodiment of the invention is determining the structure of any compound so identified.
  • the determination of the structures of compounds having the desired characteristic can be accomplished by decoding the tags (represented by T'-L- in Formula I) or, alternatively, by deconvolution of the library (Smith et al., BioMed. Chem. Lett., 4, 2821 (1994); Kurth et al,,
  • Another embodiment of the invention is the use of divinylbenzene-cross-linked, polyethyleneglycol-grafted polystyrene beads optionally functionalized with amino groups (for example, TentaGel® S NH 2 , Rapp Polymere) as the solid supports for
  • Alkyl is intended to include linear, branched, or cyclic hydrocarbon structures and combinations thereof.
  • “Lower alkyl” means alkyl groups of from 1 to 8 carbon atoms. Examples of lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s- and t-butyl, pentyl, hexyl, octyl, cyclopropylethyl, and the like.
  • “Lower cycloalkyl” includes cycloalkyl groups of from 3 to 8 carbon atoms. Examples of lower cycloalkyl groups include c-propyl, c-butyl, c-pentyl, 2-methylcyclopropyl, cyclopropylmethyl, norbornyl, and the like.
  • Alkenyl is C 2 -C 8 alkenyl of a linear, branched, or cyclic (C 5 -C 6 ) configuration and combinations thereof.
  • alkenyl groups include vinyl, allyl, isopropenyl, pentenyl, hexenyl, c-hexenyl, 1-propenyl, 2-butenyl, 2-methyl-2-butenyl, and the like.
  • Alkynyl is C 2 -C 8 alkynyl of a linear or branched configuration and combinations thereof. Examples of alkenyl groups include ethyne, propyne, butyne, pentyne, 3-methyl-1-butyne, 3,3-dimethyl-1-butyne, and the like.
  • Alkoxy means alkoxy groups of from 1 to 8 carbon atoms of a straight, branched, or cyclic configuration and combinations thereof. Examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy, and the like.
  • Acylamino means acylamino groups of from 1 to 8 carbon atoms of a straight, branched or cyclic configuration and combinations thereof. Examples of acylamino groups are acetylamino, butylamino, cyclohexylamino, and the like.
  • Hal means halogen, which includes F, Cl, Br, and I.
  • Halophenyl means phenyl substituted by 1-5 halogen atoms. Halophenyl includes pentachlorophenyl, pentafluorophenyl, and 2 ,4,6-trichlorophenyl.
  • Aryl and “heteroaryl” mean a 5- or 6-membered aromatic or heteroaromatic ring containing 0-3 heteroatoms selected from O, N, and S; a bicyclic 9- or 10-membered aromatic or heteroaromatic ring system containing 0-3 heteroatoms selected from O, N, and S; or a tricyclic 13- or 14-membered aromatic or
  • the aromatic 6- to 14-membered carbocyclic rings include benzene, naphthalene, indane, tetralin, and fluorene and the 5- to 10-membered aromatic heterocyclic rings include imidazole, pyridine, indole, thiophene, benzopyranone, thiazole, furan, benzimidazole, quinoline, isoquinoline, quinoxaline, pyrimidine, pyrazine, tetrazole, and pyrazole.
  • Substituted alkyl, alkenyl, or alkynyl means alkyl, alkenyl, or alkynyl wherein up to three H atoms on each C therein are replaced by halogen, hydroxy, loweralkoxy, carboxy, carboalkoxy, carboxamido, cyano, carbonyl, NO 2 , NR 3 R 3 , alkylthio, sulfoxide, sulfone, acylamino, amidino, phenyl, benzyl, heteroaryl, phenoxy, benzyloxy, heteroaryloxy, and substituted phenyl, benzyl, heteroaryl, phenoxy, benzyloxy, and heteroaryloxy.
  • NR 3 R 3 represents NHH, NHCH 3 , N(CH 3 ) 2 , etc.
  • the left-hand bond is the point of attachment to the solid support (via -C(O)- for L' and -C(O)- or -CH 2 C(O)- for L) and the right-hand bond is the point of attachment to either T or II.
  • the tags of this invention, T are chemical entities which possess several properties: they must be detachable from the solid supports, preferably by photolysis or oxidation; they must be
  • the tags may be structurally related or unrelated, e.g., a homologous series, repetitive functional groups, related members of the Periodic Chart, different isotopes, combinations thereof, and the like. At the end of the combinatorial synthesis, to each solid support, there will usually be attached at least 0.01 femtomol, usually 0.001-50 pmol, of each tag.
  • the tags may be aliphatic, alicyclic, aromatic, heterocyclic, or combinations thereof.
  • Distinguishing features may be the number of repetitive units, such as methylene groups in an alkyl moiety; alkyleneoxy groups in a
  • polyalkyleneoxy moiety halo groups in a polyhalo compound; ⁇ -and/or ⁇ -substituted ethylene groups where the substituents may be alkyl groups, oxy, carboxy, amino, halo, or the like; isotopes; etc.
  • solid supports The materials upon which the combinatorial syntheses of the invention are performed are referred to as solid supports, beads, and resins. These terms are intended to include:
  • beads, pellets, disks, fibers, gels, or particles such as cellulose beads, pore-glass beads, silica gels, polystyrene beads optionally cross-linked with divinylbenzene and optionally grafted with polyethylene glycol and optionally functionalized with amino, hydroxy, carboxy, or halo groups, grafted co-poly beads, poly-acrylamide beads, latex beads, dimethylacrylamide beads optionally cross-linked with N,N'-bis-acryloyl ethylene diamine, glass particles coated with hydrophobic polymer, etc., i.e., material having a rigid or semi-rigid surface;
  • soluble supports such as low molecular weight non-cross-linked polystyrene; and. c) derivatized forms thereof such as
  • Suitable aminoacid protecting groups are well known in the art and include Fmoc, Alloc (allyloxycarbonyl), etc.
  • Some of the compounds described herein contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisometric forms which may be defined in terms of absolute stereochemistry as (R) or (S), or as (D) or (L) for amino acids.
  • the present invention is meant to include all such possible diastereomers as well as their racemic and optically pure forms.
  • Optically active (R) and (S), or (D and L), isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.
  • the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended to include both E and Z geometric isomers. Likewise, all tautomeric forms are intended to be included.
  • the library of the present invention is useful as a screening tool for discovering new lead structures by evaluation across an array of biological assays, including the discovery of selective inhibition patterns across isozymes.
  • the library is thus a tool for drug discovery; i.e., as a means to discover novel lead compounds by screening the library against a variety of biological targets and to develop structure-activity relationships (SAR) in large families of related compounds.
  • the library may be tested with the ligands attached to the solid supports as depicted in Formula I or I', or the compounds II may be detached prior to evaluation. With the compounds of Formula I or I', screening assays such as FACS sorting and cell lawn assays may be used.
  • tags attached to solid support associated with bioactivity may then be decoded to reveal the structural or synthetic history of the active compound (Ohlmeyer et al., Proc. Natl. Acad. Sci. USA, 90, 10922-10926, Dec. 1993 and Still et al., Complex Combinatorial Chemical Libraries Encoded with Tags, WO 94/08051) or,
  • the structures may be determined by deconvolution.
  • the usefulness of such a library as a screening tool is demonstrated by Burbaum et al., Proc Natl. Acad. Set USA, 92, 6027-6031, June 1995, who describe the assaying of encoded combinatorial libraries for, e.g., carbonic anhydrase inhibition. Even if no compounds are found to be active in a given screen, such lack of activity often provides useful S AR information.
  • the compounds of the present invention can be prepared according to the following methods.
  • each solid support upon which a compound is being synthesized may be uniquely tagged to define the particular chemical event(s) occurring during that step.
  • the tagging is accomplished using identifiers such as those of Formula IV, which record the sequential events to which the support is exposed during the synthesis, thus providing a reaction history for the compound produced on each support.
  • the identifiers are used in combination with one another to form a binary or higher order encoding scheme permitting a relatively small number of identifiers to encode a relatively large number of reaction products. For example, when used in a binary code, N identifiers can encode up to 2 N different compounds and/or conditions.
  • each variable or combination of variables at each step of the synthesis By associating each variable or combination of variables at each step of the synthesis with a combination of identifiers which uniquely define the chosen variables such as reactant, reagent, reaction conditions, or combinations of these, one can use the identifiers to define the reaction history of each solid support.
  • the syntheses one begins with at least 10 4 , desirably at least 10 7 , and generally not exceeding 10 15 solid supports.
  • the tagging may be done prior to, concomitantly with, or after the reactions which comprise each choice.
  • sample supports may be picked at any stage and a portion of their tags detached and decoded to verify that the correct tags are bound to the sample supports.
  • one may wash the beads free of any excess reagents or by-products before proceeding.
  • the supports are combined, mixed, and again divided, this time into as many containers as pre-determined for the number of choices for the second step in the synthesis. This procedure of dividing, reacting, tagging, and remixing is repeated until the combinatorial synthesis is completed.
  • the resin is filtered and washed and then suspended in an involatile solvent (i.e., a non-protic, organic solvent with a boiling point of 80-140oC) such as toluene, xylene, or chlorobenzene and treated with an excess of a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine.
  • an involatile solvent i.e., a non-protic, organic solvent with a boiling point of 80-140oC
  • a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine.
  • Compounds 4, 8, 11, and 16 have been synthesized by the aza-Wittig method of the present invention.
  • Compound 4 is a known intermediate useful in the synthesis of antitumor antibiotics (Kaneko et al., Tet. Lett., 1983, p. 5165; Kaneko et al., J. Med. Chem., 1985, p. 388).
  • Compound 8 is a novel compound useful as an intermediate in the synthesis of the antibiotic 5-thioabbeymycin (Kamal et al., Bioorg. Med. Chem. Lett., 3, p. 743, 1993) and abbeymycin.
  • Compound 11 is a novel compound useful as an intermediate in the synthesis of antitumor antibiotics
  • Resin-linked ⁇ -amino ester, A is suspended in an aprotic, polar solvent such methylene chloride, DMF, THF or ethyl acetate.
  • an aprotic, polar solvent such as methylene chloride, DMF, THF or ethyl acetate.
  • An excess (2-50 equivalents) of a soluble organic base such as
  • triethylamine, N,N-diisopropylethylamine, or pyridine is added to the suspended resin.
  • an acylation catalyst such as 4-dimethylaminopyridine may be added.
  • This mixture is treated with an excess (2-10 equivalents) of an appropriately substituted 2-azidobenzoyl chloride and agitated at room temperature for 2-24 hr.
  • the resin is filtered and washed multiple times with an appropriate solvent such as methylene chloride to remove excess reagents and byproducts.
  • Resin linked N-(2-azidobenzoyl)amino ester, B is suspended in an involatile solvent such as toluene, xylene, or chlorobenzene and treated with an excess (2-10 equivalents) of a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine.
  • a trivalent phosphorus reagent such as triphenylphosphine or tributylphosphine.
  • This mixture is agitated and heated to 80- 140°C for 2-24 hr, then cooled and the resin-linked 1,4- benzodiazepin-5-one, C, is washed multiple times with appropriate solvents such as methylene chloride or toluene to remove excess reagents and byproducts.
  • Resin-linked 1,4-benzodiazepin-5-one, C is suspended in an acidic solution such as trifluoroacetic acid/methylene chloride (50-90% TFA/CH 2 Cl 2 ) or hydrogen chloride/dioxane (1-4M HCl/dioxane) and agitated at room temperature for 1-24 hr.
  • the resin is filtered and washed with appropriate solvents such as methylene chloride or dioxane.
  • the filtrate and washings are combined and evaporated to give the crude 1,4-benzodiazepin-2,5-dione, D, which may be purified and characterized by standard techniques.
  • n 3-12 and Ar is pentachlorophenyl or
  • n 54-6 and Ar is 2,4,6-trichlorophenyl
  • Methyl vanillate (0.729 g, 4.0 mmole), 1-hydroxy-9-(2,3,4,5,6-pentachlorophenoxy)nonane (1.634 g, 4.0 mmole) and triphenylphosphine (1,258 g, 4.8 mmole) were dissolved in 20 mL dry toluene under argon.
  • DEAD (0.76 mL, 0.836 g, 4.8 mmole) was added dropwise and the mixture was stirred at 25°C for one hr. The solution was concentrated to half volume and purified by flash chromatography eluting with DMC to give 1.0 g (1.7 mmole, 43%) of the product as a white crystalline solid.
  • Step (b) The crude material from Step (b) was dissolved in 100 mL toluene, 10 mL (1.63 g, 14 mmole ) thionyl chloride was added, and the mixture was refluxed for 90 min. The volume of the solution was reduced to approx. 30 mL by distillation, then the remaining toluene was removed by evaporation.
  • the crude acid chloride was dissolved in 20 mL dry DCM and cooled to -70°C under argon and a solution of approx. 10 mmole diazomethane in 50 mL anhydrous ether was added. The mixture was warmed to r.t. and stirred for 90 min.
  • Example 5 In the synthesis of Example 5, the 12 identifiers were used to encode the combinatorial library.
  • Step 2
  • Step 1 reagent 3 is encoded “011 " which
  • N-Fmoc-cis-4-benzoyloxy-L-proline p-alkoxybenzyl resin, 5 was prepared by routine methods as outlined below.
  • TentaGel resin may be modified with bis-Fmoc lysine to increase the available reaction sites for ligand attachment.
  • the schemes elsewhere herein do not show the use of this modification with lysine.
  • the resulting mixture was treated with 4-dimethylamino-pyridine (92 mg, 0.75 mmol, 0.1 equiv.), followed by N,N'-diisopropylcarbodiimide (4.73 g, 37.5 mmol, 5 equiv.), and the reaction mixture was shaken at ambient temperature. After 6 hours, the solvent was removed by filtration, and the beads were washed successively with 5 ⁇ 150 mL DMF, and 5 ⁇ 150 mL methylene chloride. A small portion of the resin was checked by the Kaiser test for disappearance of free NH2, and found to be negative.
  • the resin was treated with a 30% solution of piperidine in DMF (100 mL), and shaken at 25°C for 1 hr.
  • the resin was filtered and washed successively with 5 ⁇ 150 mL DMF and 5 ⁇ 150 mL methylene chloride A small portion of the resin was checked with the Kaiser test to assure removal of the Fmoc groups, and found to be positive.
  • the acetate protected resin from above was treated with a solution of 10% hydrazine hydrate in methanol (100 mL), and shaken at ambient temperature. After 6 hr., the solvent was removed by filtration, and the resin was washed successively with 5 ⁇ 150 mL MeOH and then re-treated with 10% hydrazine hydrate in MeOH (100 mL) and shaken at 25oC for 16 hr. The solvent was removed by filtration, and the resin was washed successively with 5 ⁇ 150 mL DMF and 5 ⁇ 150 mL CH 2 CI 2 to afford the hydroxy resin A.
  • N-Fmoc-L-proline p-alkoxybenzyl resin, 1, (Bachem) (0.34 mmole/g, 2.0 g, 0.68 mmole) was suspended in 30 mL DMF, then filtered. The resin was then shaken with 50% piperidine/DMF for 2 hr. then filtered and washed with DMF (5 ⁇ 30 mL) and methylene chloride (10 ⁇ 30 mL). The resin was suspended in 20 mL methylene chloride, and 2 mL (14 mmole) triethylamine was added, followed by 1.0 g (5.5 mmole, 8 equiv.) 2-azidobenzoyl chloride.
  • the mixture was shaken at 25°C for 3 hr. and then the resin was washed with methylene chloride (10 ⁇ 30 mL) followed by toluene (10 ⁇ 30 mL).
  • the resin, 2 was suspended in 20 mL toluene and 0.3 g (1.14 mmole, 1.7 equiv.) triphenylphosphine was added and the mixture was shaken until the triphenylphosphine had dissolved.
  • the mixture was shaken and heated to 80°C for 3 hr. then cooled and washed with methylene chloride (10 ⁇ 30 mL).
  • the pale brown resin, 3, was dried under vacuum.
  • the resin, 3 (2.0 g, 0.68 mmole), prepared as described above, was suspended in 20 mL methylene chloride and 20 mL trifluoroacetic acid was added. The resin was shaken at 25°C for 30 min then filtered and washed with methylene chloride (2 ⁇ 20 mL). The filtrate and washings were collected and combined, then evaporated to give the crude product.
  • the benzodiazepine was purified by flash chromatography, eluting with 60% ethyl acetate/hexane to give the product, 4, as a white solid.
  • N-Fmoc-cis-4-benzoyloxy-L-proline p-alkoxybenzyl resin, 5, (400 mg) was suspended in 10 mL DMF, then filtered. The resin was then shaken with 50% piperidine/DMF, 10 mL, for 2 hr. then filtered and washed with DMF (5 ⁇ 10 mL) and methylene chloride (10 ⁇ 10 mL). The resin was suspended in 10 mL methylene chloride, 0.4 mL ( ⁇ 3 mmole) triethylamine was added, followed by 0.2 g (1.1 mmole) 2-azido-benzoyl chloride. The mixture was shaken at 25°C for 12 hr.
  • N-Fmoc-L-proline p-alkoxybenzyl resin, 1, (0.34 mmole/g, 5.0 g, 1.8 mmole) was suspended in 50 mL DMF, then filtered. The resin was then shaken with 50% piperidine/DMF for 2 hr. then filtered and washed with DMF (5 ⁇ 50 mL) and methylene chloride (10 ⁇ 50 mL). The resin was suspended in 50 mL methylene chloride, 3 mL (21 mmole) triethylamine was added, followed by 0.75 g (3 mmole, 1.7 equiv.) 2-azido-4,5-dimethoxybenzoyl chloride as a solid.
  • the mixture was shaken at 25 °C for 12 hr. and then the resin was washed with methylene chloride (10 ⁇ 50 mL), followed by toluene (10 ⁇ 50 mL).
  • the resin, 9, was suspended in 50 mL toluene and 1.26 g (4.8 mmole 2.6 equiv.) triphenylphosphine was added and the mixture was shaken until the triphenylphosphine had dissolved.
  • the mixture was shaken and heated to 80°C for 2 hr. then cooled and washed with toluene (10 ⁇ 50 mL), alternately with methanol and methylene chloride (5 x 50 mL each) and finally methylene chloride (5 ⁇ 50 mL).
  • the pale brown resin, 1 was dried under vacuum.
  • p-Alkoxybenzyl alcohol resin 12, 2.5 g (1.0 mmole/g) was suspended in 30 mL methylene chloride. N-Fmoc-N-methyl-L-phenylalanine, 3.0 g (7.5 mmole, 3 equiv.), DMAP, 1.0 g (0.82 mmole, 0.3 equiv.), and DIC 1.74 mL (1.26 g, 10 mmole, 4 equiv.) were added and the mixture was agitated for 2 hr. The resin was filtered and washed with methylene chloride, (10 ⁇ 30 mL), then dried.
  • the mixture was shaken at 25 C for 1 hr and then the resin was washed with methylene chloride (10 ⁇ 30 mL) followed by toluene (10 ⁇ 30 mL).
  • the resin, 14, was suspended in 20 mL toluene and 0.52 g (2.0 mmole) triphenylphosphine was added and the mixture was shaken until the triphenylphosphine had dissolved.
  • the mixture was shaken and heated to 90oC for 3 hr then cooled and washed with toluene (5 ⁇ 20 mL) and methylene chloride (10 ⁇ 20 mL) to give the resin linked 1,4-benzodiazepine-5-one, 15.
  • the resin, 15, was suspended in 10 mL methylene chloride and 10 mL trifluoroacetic acid was added. The resin was shaken at 25°C for 2 hr, then filtered and washed with methylene chloride (2 ⁇ 20 mL). The filtrate and washings were collected and combined, then evaporated to give the crude product which was purified by flash chromatography eluting with 50% ethyl acetate/hexane to give the product, 16.
  • the resin above was divided into two equal batches of 1.75 g which were placed into two separate 100 mL synthesis vessels (vessels 1 and 2) and sixty equal batches of 0.85 g which were placed into sixty separate 100 mL synthesis vessels (vessels 3-62).
  • the 62 resin batches were encoded with six tags as follows.
  • (iv) The resin batches were suspended in 30 mL methylene chloride. Aliquots, 2 mL of 37 mg/mL (74 mg tag precursor/vessel -8.7 % by mass of resin), of C 6 CI 5 tag precursor solution in methylene chloride were added to the appropriate vessels from 3 to 62 and the vessels were shaken for 2 min. A 3 mL aliquot of 37 mg/mL C 6 CI 5 tag precursor solution was added to vessel 1 (111 mg tag precursor in vessel 1, 6.3% by mass of resin). Aliquots, 2 mL of 37 mg/mL (74 mg tag
  • a collection of 51 319 1,4-benzodiazepine-2,5-diones was prepared as follows. Resin batches 3C-62C were combined and mixed thoroughly in a seperatory funnel, then divided as a slurry in methylene chloride into nineteen equal portions (-2.7 g, 1.4 mmole) in 100 mL synthesis vessels. The resin was suspended in 30 mL methylene chloride and diisopropylethylamine, 2.5 mL (1.8 g, 14 mmole, 10 equiv), was added followed by 5 mmole of the appropriate o-azidobenzoyl chloride (Reagent Set 3).
  • Tributylphosphine 1.8 mL (1.45 g, 7.2 mmole, 5 equiv), was added to each flask and the mixtures were heated to 140-150°C for 6 hr, then cooled, filtered, and washed with 2 ⁇ 30 mL toluene and 5 ⁇ 30 mL methylene chloride to give the resin linked benzodiazepines, E.
  • the product 1,4-benzodiazepine-2,5-diones, F were cleaved from the resin support by suspending the resin, 20 beads, in 100 ⁇ L of 70 % TFA/water for 4 hours and filtering the solution.
  • a second collection of 1 170 1,4-benzodiazepine-2,5-diones was prepared as follows. Resin batches 1C and 2C were combined and mixed thoroughly in a seperatory funnel. The resin was dried in vacuo and divided into thirteen equal portions ( ⁇ 0.27 g, 0.14 mmole) then placed in 20 mL synthesis vessels. The resin was suspended in 10 mL methylene chloride and diisopropylethylamine, 0.25 mL (0.18 g, 1.4 mmole, 10 equiv), was added followed by 0.5 mmole of thirteen o-azidobenzoyl chlorides. The mixtures were shaken at room
  • Tributylphosphine 0.14 mL (0.11 g, 0.55 mmole, 4 equiv), was added to each flask and the mixtures were heated to 140-150°C for 6 hr, then cooled, filtered, and washed with 2 ⁇ 10 mL toluene and 5 ⁇ 10 mL methylene chloride to give the resin batches E, from which the product 1,4-benzodiazepine-2,5-diones, F, may be cleaved as described above.
  • a bead is placed in a 1.3 mm diameter pyrex capillary with
  • the GC analysis is performed with a Hewlett Packard 5890 plus gas chromatograph. On column injection into a 5 m, 0.32 mm retention gap connected to a 25 m, 0.2 mm crosslinked 5%
  • phenylmethyl silicone column is used.
  • the temperature and pressure programs for the analysis are 200-320°C, 15°C/min, then 320°C for 10 min and 20-40 psi at 2 psi/min, then 40 psi for 10 min.
  • the EC detector is maintained at 400°C and the auxiliary gas is set at 35 psi.
  • the identity of the library compound attached to the bead is ascertained based on the reagents utilized in the synthesis of such compound, which are readily determined from the binary codes associated, respectively, with each of the identifiers for such reagents, as characterized through the above procedure.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Indole Compounds (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

La présente invention concerne un procédé nouveau permettant de synthétiser une banque combinatoire de 1,4-benzodiazépine-2,5-diones sur des supports solides par l'intermédiaire de la fermeture d'un cycle d'aza-Wittig. Les composés en question peuvent, le cas échéant, être encodés avec des étiquettes. L'invention concerne également l'utilisation de cette banque dans des analyses pour découvrir des composés actifs dans le domaine biologique et, en option, en séparer les 1,4-benzodiazépine-2,5-diones.
PCT/US1996/011070 1995-06-29 1996-06-28 Banque combinatoire de 1,4-benzodiazepine-2,5-diones Ceased WO1997001560A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP96923532A EP0850238A4 (fr) 1995-06-29 1996-06-28 Banque combinatoire de 1,4-benzodiazepine-2,5-diones
AU64020/96A AU6402096A (en) 1995-06-29 1996-06-28 Combinatorial 1,4-benzodiazepin-2,5-dione library
JP9504592A JPH11508563A (ja) 1995-06-29 1996-06-28 １，４−ベンゾジアゼピン−２，５−ジオン組合わせライブラリー
US08/996,579 US5962337A (en) 1995-06-29 1997-12-23 Combinatorial 1,4-benzodiazepin-2,5-dione library

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US65795P 1995-06-29 1995-06-29
US60/000,657 1995-06-29

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US08/996,579 Continuation-In-Part US5962337A (en) 1995-06-29 1997-12-23 Combinatorial 1,4-benzodiazepin-2,5-dione library

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EP (1) EP0850238A4 (fr)
JP (1) JPH11508563A (fr)
AU (1) AU6402096A (fr)
CA (1) CA2225914A1 (fr)
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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000012506A3 (fr) * 1998-08-27 2000-06-29 Univ Portsmouth Collections de composes
US6136295A (en) * 1996-05-24 2000-10-24 Massachusetts Institute Of Technology Aerodynamically light particles for pulmonary drug delivery
US6747144B1 (en) 1998-08-27 2004-06-08 Spirogen Limited Collections of compounds
WO2003093823A3 (fr) * 2002-05-01 2004-07-22 Pharmacopeia Inc Etiquettes moleculaires chimiquement inertes
US6909006B1 (en) 1999-08-27 2005-06-21 Spirogen Limited Cyclopropylindole derivatives
US6939712B1 (en) 1998-12-29 2005-09-06 Impedagen, Llc Muting gene activity using a transgenic nucleic acid
US7049311B1 (en) 1998-08-27 2006-05-23 Spirogen Limited Pyrrolbenzodiazepines
US7052678B2 (en) 1997-09-15 2006-05-30 Massachusetts Institute Of Technology Particles for inhalation having sustained release properties
US7407951B2 (en) 2002-11-14 2008-08-05 Spirogen Limited Pyrrolobenzodiazepines
US7429658B2 (en) 2003-09-11 2008-09-30 Spirogen Limited Synthesis of protected pyrrolobenzodiazepines
EP1948191A4 (fr) * 2005-11-01 2011-01-05 Univ Michigan Nouvelles 1,4-benzodiazépine-2,5-diones dotées de propriétés thérapeutiques
US9126978B2 (en) 2009-11-17 2015-09-08 The Regents Of The University Of Michigan 1,4-benzodiazepine-2,5-diones and related compounds with therapeutic properties

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5191404A (en) * 1989-12-20 1993-03-02 Digital Equipment Corporation High density memory array packaging

Citations (2)

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Publication number Priority date Publication date Assignee Title
US3173912A (en) * 1961-10-02 1965-03-16 Olin Mathieson Benzodiazepines
US3860600A (en) * 1972-07-10 1975-01-14 Sterling Drug Inc Octahydropyrrido{8 2,1-c{9 {8 1,4{9 benzodiazepines

Family Cites Families (1)

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Publication number Priority date Publication date Assignee Title
ATE244769T1 (de) * 1992-10-01 2003-07-15 Univ Columbia Komplexe kombinatorische chemische banken, die mit markierungen versehen sind

Patent Citations (2)

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Publication number Priority date Publication date Assignee Title
US3173912A (en) * 1961-10-02 1965-03-16 Olin Mathieson Benzodiazepines
US3860600A (en) * 1972-07-10 1975-01-14 Sterling Drug Inc Octahydropyrrido{8 2,1-c{9 {8 1,4{9 benzodiazepines

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0850238A4 *

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6136295A (en) * 1996-05-24 2000-10-24 Massachusetts Institute Of Technology Aerodynamically light particles for pulmonary drug delivery
US7052678B2 (en) 1997-09-15 2006-05-30 Massachusetts Institute Of Technology Particles for inhalation having sustained release properties
US7704924B2 (en) 1998-08-27 2010-04-27 Spirogen Limited Library of compounds comprising pyrrolobenzodiazepine moieties
US6608192B1 (en) 1998-08-27 2003-08-19 Spirogen Limited Collections of compounds
US6747144B1 (en) 1998-08-27 2004-06-08 Spirogen Limited Collections of compounds
WO2000012506A3 (fr) * 1998-08-27 2000-06-29 Univ Portsmouth Collections de composes
US7049311B1 (en) 1998-08-27 2006-05-23 Spirogen Limited Pyrrolbenzodiazepines
US7067511B2 (en) 1998-08-27 2006-06-27 Spirogen Limited Pyrrolobenzodiazepines
US7265105B2 (en) 1998-08-27 2007-09-04 Spirogen Limited Pyrrolobenzodiazepines
US6939712B1 (en) 1998-12-29 2005-09-06 Impedagen, Llc Muting gene activity using a transgenic nucleic acid
US6909006B1 (en) 1999-08-27 2005-06-21 Spirogen Limited Cyclopropylindole derivatives
WO2003093823A3 (fr) * 2002-05-01 2004-07-22 Pharmacopeia Inc Etiquettes moleculaires chimiquement inertes
US7407951B2 (en) 2002-11-14 2008-08-05 Spirogen Limited Pyrrolobenzodiazepines
US7429658B2 (en) 2003-09-11 2008-09-30 Spirogen Limited Synthesis of protected pyrrolobenzodiazepines
EP1948191A4 (fr) * 2005-11-01 2011-01-05 Univ Michigan Nouvelles 1,4-benzodiazépine-2,5-diones dotées de propriétés thérapeutiques
EP2604269A1 (fr) * 2005-11-01 2013-06-19 The Regents Of The University Of Michigan 1,4-benzodiazepine-2,5-diones possédant des propriétés thérapeutiques
US9126978B2 (en) 2009-11-17 2015-09-08 The Regents Of The University Of Michigan 1,4-benzodiazepine-2,5-diones and related compounds with therapeutic properties
US9849138B2 (en) 2009-11-17 2017-12-26 The Regents Of The University Of Michigan 1,4-benzodiazepone-2,5-diones and related compounds with therapeutic properties

Also Published As

Publication number Publication date
EP0850238A4 (fr) 1998-10-07
AU6402096A (en) 1997-01-30
EP0850238A1 (fr) 1998-07-01
CA2225914A1 (fr) 1997-01-16
JPH11508563A (ja) 1999-07-27

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