WO1995032425A1 - Bibliotheques combinatoires codees - Google Patents

Bibliotheques combinatoires codees Download PDF

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Publication number
WO1995032425A1
WO1995032425A1 PCT/US1995/006392 US9506392W WO9532425A1 WO 1995032425 A1 WO1995032425 A1 WO 1995032425A1 US 9506392 W US9506392 W US 9506392W WO 9532425 A1 WO9532425 A1 WO 9532425A1
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Prior art keywords
beads
library
reaction
combinatorial
encoded
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Dennis Shinji Yamashita
Joseph Weinstock
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SmithKline Beecham Corp
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SmithKline Beecham Corp
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Priority to JP7530459A priority Critical patent/JPH10500951A/ja
Priority to US08/537,752 priority patent/US6210900B1/en
Priority to EP95920576A priority patent/EP0763202A4/fr
Publication of WO1995032425A1 publication Critical patent/WO1995032425A1/fr
Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/005—Beads
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/0054—Means for coding or tagging the apparatus or the reagents
    • B01J2219/00572—Chemical means
    • B01J2219/00576—Chemical means fluorophore
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00592—Split-and-pool, mix-and-divide processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00596—Solid-phase processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/10—Libraries containing peptides or polypeptides, or derivatives thereof

Definitions

  • the field of this invention concerns combinatorial chemistry which involves the syntheses of one or more encoded combinatorial libraries where large numbers of products having varying compositions are obtained. This invention also relates to methods of encoding combinatorial libraries.
  • the standard method for conducting a search is to screen a variety of pre-existing chemical moieties, for example, naturally occurring compounds or compounds which exist in synthetic libraries or databanks.
  • the biological activity of the pre-existing chemical moieties is determined by applying the moieties to an assay which has been designed to test a particular property of the chemical moiety being screened, for example, a receptor binding assay which tests the ability of the moiety to bind to a particular receptor site.
  • Nonpeptidic organic compounds such as peptide mimetics
  • peptide mimetics can often surpass peptide ligands in affinity for a certain receptor of enzyme.
  • An effective strategy for rapidly identifying high affinity biological ligands, and ultimately new and important drugs, requires rapid construction and screening of diverse libraries of non-peptidic structures containing a variety of structural units capable of establishing one or more types of interactions with a biological acceptor (e.g., a receptor or enzyme), such as hydrogen bonds, salt bridges, pi-complexation, hydrophobic effects, etc.
  • a biological acceptor e.g., a receptor or enzyme
  • a key unsolved problem in the area of generation and use of nonpeptide libraries is the generation and use of nonpeptide libraries is the elucidation of the structure of molecules selected from a library that show promising biological activity.
  • An attempt to uncover the structures of peptides selected from a library using unique nucleotide sequence codes, which are synthesized in tandem with the peptide library, has been described by Brenner and Lerner (Brenner, S. and Lerner, R.A. Proc. Nat'l. Acad. Sci. USA, 1992 89 . 5381-5383).
  • the nucleotide sequence of the code attached to each peptide must be amplifiable via the polymerase chain reaction (PCR).
  • nucleotide synthesis techniques are not compatible with all of the synthetic techniques required for synthesis of many types of molecular libraries.
  • the close proximity of nucleotide and synthetic test compound in the library which can result in interactions between these molecules interfering with the binding of the ligand with a target receptor of enzyme during the biological assay, also limits this approach.
  • the nucleotide component of the library can also interfere during biological assays in a variety of other ways.
  • Kerr et al. J, Am, Chem, Soc., 1993, 115, 2520-2531
  • the peptide ligand and its coding strand in this library are covalently joined together, which allows isolation and sequence determination of pairs of synthetic test compound and corresponding code.
  • the coding peptide may interfere with the screening assay.
  • PCT/US93/09345 describes a method of identifying actives in a
  • combinatorial library by attaching multiple tags in a predetermined binary coding system.
  • PCT/HU93/0030 describes fluorescently labeled sub-library peptide kits for use in peptide synthesis.
  • PCT/US94/06078 describes methods of encoding combinatorial libraries using polymeric sequences.
  • This invention relates to a method for identifying compounds having desired characteristics and identifying essential moieties in a lead structure which comprises preparing one or more encoded combinatorial libraries from a specified set of reaction sequences and testing compounds therein for biological activity.
  • This invention also relates to a method of encoding a single registry in each combinatorial library of a series of combinatorial libraries and combinatorial libraries with a single encoded registry.
  • This invention also relates to a method of encoding combinatorial libraries which comprises utilization of tagged beads.
  • This invention also relates to a method of encoding each choice of a combinatorial library and combinatorial libraries encoded thereby.
  • This invention also relates to beads with fluorescently labeled identifiers attached thereto.
  • beads means any solid support material capable of providing a base for combinatorial syntheses and capable of being processed by flow cytometry, such as 1 to 2% crosslinked polystyrene, polyacrylamide, polyethylene glycol polystyrene co-polymer, preferably Tentagel 10 to 100 micron particles, most preferably Tentagel 10-30 micron particles.
  • sort means to form beads into groups which have a common tagging aspect by flow cytometry.
  • the term "separate” or “split” when referring to encoded beads or beads of a combinatorial library means to partition the mixture of beads into groups, each group thereinby containing a mixture, preferably a statistical mean of all members.
  • the term "tag”, unless otherwise indicated, means an encoding characteristic of a bead or group of beads which is capable of being sorted by flow cytometry, such as differences in size, differences in material composition, differences in flow properties, a single fluorescent marker or, preferably, a fluorescent label identifier.
  • fluorescent label identifier or "identifier” means a coding label attached to a bead or group of beads either by adding ratios of a fluorophore and a non-fluorophore or by adding multiple, preferably two, different fluorophores in varying ratios.
  • the term "intensity-differentiated” means an identifier (as used herein) in which varying ratios of a fluorophore and a non-fluorophore are added to a bead or group of beads.
  • the term "choice” means the alternative variables for a given stage in a combinatorial synthesis (not limited to peptide chemistry), such as reactant, reagent, reaction conditions, and combinations thereof.
  • stage corresponds to a step in the sequential synthesis of a compound or ligand; the compound or ligand being the final product of a combinatorial synthesis.
  • registration has the same meaning as the term "stage” as indicated above.
  • a series of combinatorial libraries are prepared, each individual library being prepared from substantially the same specified set of reaction sequences, therein encoding a single registry within each combinatorial library and analyzing according to mixtures of compounds with a homogeneous registry.
  • the specific encoded registry of any library will be different from the other libraries and the number of libraries prepared will equal the number of registries in a single library.
  • the number of readily identifiable groups of beads will correspond to the number of choices in the first registry, the entirety of each group is entered into a separate container.
  • the beads will usually be divided up into groups of at least one bead each, usually a plurality of beads, generally 1000 or more, and may be 10 5 or more depending on the total number of registries involved in the library.
  • the same reaction may be carried out in 2 or more containers to enhance the proportion of product having a particular reaction in a particular registry as compared to the other choices.
  • one or more of the registries may involve a portion of the beads being set aside and undergoing no reaction, so as to enhance the variability associated with the final product.
  • batches may be taken along different synthetic pathways. The library thus prepared will contain tagged beads which identify the reaction sequence of the first registry only.
  • a combinatorial library containing tagged beads which identify the reaction sequence of the first registry only can be prepared as outlined in Scheme 1 below.
  • Scheme 1 outlines the preparation of a combinatorial library in which only the first registry has been encoded.
  • beads with attached fluorescently labeled identifiers are derivatized with a linker that allows for cleavage of the compound to be tested.
  • each group of similarly tagged beads is entered into a separate container and subjected to specified reaction conditions (or variable building blocks, as used herein) to form the first registry.
  • specified reaction conditions or variable building blocks, as used herein
  • the beads In carrying out the synthesis to prepare the second library, one will preferably begin with the same number of beads as used in the first library, said beads may be tagged in a similar manner as in the first library.
  • the beads for use in the second library are first combined into a single mixture and then separated according to the number of choices for the first registry.
  • schemeSchoices for each registry of the second library and all subsequent libraries will be substantially the same as the synthetic schemeSchoices of the corresponding registry in the first library.
  • the reaction(s) may wish to wash the beads free of any reagent, followed by combining all of the beads into a single mixture and then separating the beads according to the number choices in the third registry of the first library. This procedure of dividing beads, followed by the synthetic scheme(s) ⁇ choice(s) from the corresponding registry of the first library, and then recombining the beads is iterated until the second library in completed.
  • the library thus prepared will contain tagged beads which identify the reaction sequence of the second registry only.
  • a combinatorial library containing tagged beads which identify the reaction sequence of the second registry only can be prepared as outlined in Scheme 2 below.
  • step 4 Deprotect FMOC; couple FMOC-NHCHR B1-BN CO 2 H. 5) Combine and separate. 6) Repeat step 4 and 5, except replace FMOC-NHCHR B 1-BN CO 2 H with FMOC-NHCHR C1-CN CO 2 H, ... FMOC-NHCHR X1-XN CO 2 H until the synthesis is complete.
  • Scheme 2 outlines the preparation of a combinatorial library in which only the second registry has been encoded.
  • beads with attached fluorescent label identifiers are first combined into a single mixture and then separated into groups according to the number of choices in the first registry of the first library . Subsequently, each group is entered into a separate container and subjected to the same reaction conditions of the first registry of the first library to form the first registry of the second library. Once the reaction(s) is complete the beads are combined into a single mixture and then sorted into groups according to similarly tagged beads. Preferably this combination of beads will be sorted using flow cytometry.
  • Each group of similarly tagged beads is entered into a separate container and subjected to the same reaction conditions of the second registry of the first library to form the second registry of the second library.
  • the beads are combined into a single mixture and then separated according to the number of choices in the third registry of the first library and reacted accordingly.
  • This procedure of dividing the beads, fo by subjection to specified reaction conditions from the corresponding ⁇ of the first library, and then recombining the beads is iterated until the second horary is completed.
  • the completed library is then tested for biological activity. Information on the relative activities of mixtures of the compounds with a homogeneous second registry is obtained from this library.
  • the above process is repeated to prepare subsequent libraries (when desired), provided that the sorting procedure is performed prior to a different synthetic stage in each library.
  • the combinatorial libraries thus prepared will contain tagged beads which identify the reaction sequence of a single registry only. Further, the identifiable ⁇ encoded registry in each combinatorial library will be different. Subsequent Combinatorial Libraries
  • each library is tested separately for biological activity.
  • testing for biological activity or "testing for desired
  • the compounds of a library may be tested on the beads, for example by bio-panning using a soluble receptor assay, and the activities analyzed preferably by flow cytometry.
  • the contents of the library may be sorted preferably by flow cytometry and the compounds tested on the beads, or the sorted compounds cleaved from the beads prior to testing.
  • Analysis of the first combinatorial library will yield the SAR of variable building block A.
  • Analysis of the second combinatorial library will yield the SAR of variable building block B.
  • Analysis of the third combinatorial library will yield the SAR of variable building block C.
  • Analysis of the SARs of the three variable building blocks (A, B and C) will identify desired reaction sequences and suggest multiple lead structures.
  • fluorescent label identifiers when referring to
  • flow cytometers are able to sort beads that differ in fluorescence intensity by a factor of 2.
  • the principles of flow cytometry and general methods for using flow cytometry are described in Grogan and Collins, Guide to Flow Cytometry Methods, Pub: Marcel Dekker, Inc. (1990).
  • intensity-differentiated fluorophore-labeled beads can be prepared by the method outlined in Scheme 3 below and in the Examples.
  • R is a fluorescent tag T 1 or a doping agent
  • D and R' is a fluorescent tagT 2 or a doping agent D, provided that when R is D, R' is other that D.
  • a sample of beads is derivatized with a linker, preferably ⁇ -Boc-FMOC lysine, by standard coupling chemistry.
  • a linker preferably ⁇ -Boc-FMOC lysine
  • a benzyl alcohol linker such as used with the Wang linker or a benzyl halide linker such as used with the Merrifield linker, or a benzhydryl amine linker as used with the Rink linker can be attached to the beads by the formation of ethers by alkylation of alcohols, alkylation or arylation by Friedl Crafts chemistry, the formation of biaryls by palladium mediated cross-coupling chemistry or by standard amide coupling chemistry.
  • a mono- deprotection step such as 20% piperdine/ DMF, for removal of an FMOC is performed.
  • the beads are then divided into N pools.
  • Pool 1 is derivatized with a fluorophore, such as pyrene butyric acid.
  • Pool 2 is derivatized with a 1:3 mixture of a fluorophore, such as pyrene butyric acid, and a non-fluorophore (hereinafter a "doping agent"), such as butyric acid or a different fluorophore, such as perylene butyric acid.
  • a fluorophore such as pyrene butyric acid
  • a non-fluorophore hereinafter a "doping agent”
  • Pool N is derivatized with a 1: 3 (N-1) ratio of a fluorophore, such as pyrene butyric acid, and a doping agent, such as butyric acid or a different fluorophore, such as perylene butyric acid.
  • a fluorophore such as pyrene butyric acid
  • a doping agent such as butyric acid or a different fluorophore, such as perylene butyric acid.
  • Each of these pools of beads can be differentiated from any other pool of beads by flow cytometry.
  • Each pool of beads may also be differentiated from one another by inspection with the unaided eye, however fewer variables could be encoded this way.
  • different fluorophores with different absorption and emittance wavelengths and multiple fluorophores could be encoded by fluorescence quenching to encode additional variables.
  • the use of multiple fluorophores, the ratio of which is the identifier has several advantages including the ability to greatly increases the number of variables that can be identified by using the same number of tags and enabling analysis independent of bead size.
  • the same strategy can be applied to prepare beads that can be used to discriminate between library members with redundant molecular weights by fluorescence, preferably by starting with beads with at least 50 pmoles of linker.
  • a single combinatorial library is prepared, each choice therein being encoded by a tag, preferably using fluorescent label identifiers, and tested for biological activity, preferably without mixing the final pools.
  • the “Combine and Split protocol” is utilized to synthesize encoded beads, preferably with fluorescent label identifiers attached thereto.
  • the “Combine and Split protocol” is advantageous in that it eliminates the need to resynthesize, or parallel synthesize, libraries containing only one or two fluorescent tags. This aspect of the invention is especially attractive from a practical point of view since the encoded beads can be prepared in bulk, prior to the actual synthesis of combinatorial libraries.
  • An additionally preferred aspect of this invention relates to combinatorial libraries prepared using beads encoded by fluorescent label identifiers and to pharmaceutically active compounds identified by such combinatorial library.
  • An additionally preferred aspect of this invention relates to combinatorial libraries in which each choice therein is encoded by fluorescent label identifiers and to pharmaceutically active compounds identified by such combinatorial library.
  • An additionally preferred aspect of this invention relates to combinatorial libraries prepared using beads encoded by fluorescent label identifiers, wherein said beads were obtained by the Combine and Split protocol, and to pharmaceutically active compounds identified by such combinatorial library.
  • An additionally preferred aspec his invention relates to combinatorial libraries in which each choice therein is encoded by fluorescent label identifiers, wherein said beads were obtained by the Combine and Split protocol, and to pharmaceutically active compounds identified by such combinatorial library.
  • Y 1 is encoded by Y 2 is encoded by Y 3 is encoded by T 1c T 2c T 3c
  • the above T 1 a Registry The above T 2a Registry
  • the above T 3a Registry can be described as can be described as can be described as can be described as can be described as
  • Z 1 is encoded by Z 2 is encoded by Z 3 is encoded by T 1a T 2a T 3a
  • T 1 b P 2 is encoded by T 2b
  • Scheme 4 outlines the preparation of a combinatorial library in which each choice therein is encoded by a unique identifier.
  • untagged beads are encoded with the first identifier (as described in Scheme 3).
  • the encoded beads are combined into a single mixture and then separated into groups according to the number of permutations of the second identifier.
  • the beads are then encoded with the second identifier. (The above encoding process is repeated until groups of encoded beads of desired size is obtained).
  • the beads encoded with the second identifier are combined into a single mixture and then separated into groups according to the number of permutations of the third identifier.
  • the beads are then encoded with the third identifier.
  • Encoded beads prepared according to the above methods and said methods represent preferred embodiments of the claimed invention.
  • the beads thus prepared are maintained in separate homogeneous pools of like identifiers according to the third identifier and subjected to the first stage (or registry as used herein) of specified reaction conditions.
  • the choices of the first registry are thereinby encoded by the third identifier.
  • the beads are then combined and sorted, preferably by flow cytometry, into homogeneous pools of like identifiers according to the first identifier.
  • the beads thus obtained are maintained in separate pools and subjected to the second stage of specified reaction conditions.
  • the choices of the second registry are thereinby encoded by the first identifier.
  • the beads are then combined and sorted, preferably by flow cytometry, into
  • the beads thus obtained are maintained in separate pools and subjected to the third stage of specified reaction conditions.
  • the choices of the third registry are thereinby encoded by the second identifier.
  • the beads are then combined and separated into groups according to the number of choices of the forth stage and subjected to the forth stage of specified reaction conditions.
  • the pools of beads thus obtained are maintained in these separate groups and tested for biological activity.
  • the choices of the forth registry are thereinby separately maintained.
  • each of these groups are separately tested for biological activity and analyzed, preferably by flow cytometry or by cleavage of compounds from individual groups or from smaller sets of individual groups.
  • the exact reaction history of each active can be identified by reading the unique identifier from the corresponding bead.
  • an active compound is found in group 2 then one could analyze the individual bead by fluorescence detection. If T 3c , T 2a , T 2b were present on the bead, then the reaction history of the active structure is: Y 3 -Z 2 -P 2 -Q 2 .
  • a bifunctional linker such as e-Boc-FMOC-L- lysine (8.4 g, 6 eq., 18 mmol, Novabiochem), an amide coupling agent such as diisopropyl carbodiimide ( 2.3 g, 2.8 ml, 6 eq., 18 mmol, Aldrich) is added to Polyethylene glycol-linked to cross-linked polystyrene beads (Tentagel M NH 2 , 10 micron particle size, 15.0 g, 3 mmol, Rapp Polymere) suspended in a suitable solvent such as N-methyl pyrrolidine (300 ml) and is agitated overnight The reaction is filtered through a glass frit under aspirator pressure and is washed with DMF (5 x 100 ml).
  • a suitable solvent such as N-methyl pyrrolidine
  • the lysine derivatized beads (5.0 g), as described in Procedure A, are suspended in N-methyl pyrrolidine (100 ml), then a fluorophore such as 1-pyrene butyric acid (1.7 g, 6 eq., 6 mmol, Aldrich) and diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) are added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • the beads are then agitated in 25% TFA/CH 2 CI 2 (100 ml) for 2 h removing the Boc protective group.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried.
  • the beads are then reacted with a linker group such as the t-butyl dimethyl silyl ether of 4-(methyl hydroxy-phenyl) acetic acid ( 1.3 g, 6 mmol), diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) in N-methyl pyrrolidine (100 ml) overnight.
  • a linker group such as the t-butyl dimethyl silyl ether of 4-(methyl hydroxy-phenyl) acetic acid ( 1.3 g, 6 mmol), diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) in N-methyl pyrrolidine (100 ml) overnight.
  • the reaction is filtered through a glass frit under aspirator pressure and is washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 100 ml), then air dried.
  • the beads are then re suspended in THF ( 100 ml), and a desilylating agent such as tetrabutyl ammonium fluoride(6 ml, 1.0 M solution, 6 mmol, Aldrich) / ammonium acetate (0.92 g, 12 mmol) is used to deprotect the silyl ether producing the desired benzyl alcohol derivative.
  • a desilylating agent such as tetrabutyl ammonium fluoride(6 ml, 1.0 M solution, 6 mmol, Aldrich) / ammonium acetate (0.92 g, 12 mmol) is used to deprotect the silyl ether producing the desired benzyl alcohol derivative.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried.
  • the beads ( 5 g), as prepared in Procedure B, are then suspended in N- methyl pyrrolidine (100 ml) and is then reacted with a monomer such as FMOC-L- glyine (1.8 g, 6 eq., 6 mmol).
  • a monomer such as FMOC-L- glyine (1.8 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • Procedure D The lysine derivatized beads (5.0 g), as described in Procedure A, are suspended in N-methyl pyrrolidine (100 ml), then a fluorophore such as 1-pyrene butyric acid (0.43 g, 1.5 eq., 1.5 mmol), and a doping agent such as butyric acid (0.4 g, 0.41 ml, 4.5 eq., 4.5 mmol) in 1:3 stoichiometry, and diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) are added, and the reaction is agitated for 3 hours.
  • a fluorophore such as 1-pyrene butyric acid (0.43 g, 1.5 eq., 1.5 mmol)
  • a doping agent such as butyric acid (0.4 g, 0.41 ml, 4.5 eq., 4.5 mmol) in 1:3 stoichiometry
  • reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • the beads are then agitated in 25% TFA/ CH 2 CI 2 (100 ml) for 2 h removing the Boc protective group.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried.
  • the beads are then reacted with a linker group such as the t-butyl dimethyl silyl ether of 4-(methyl hydroxy-phenyl) acetic acid ( 1.3 g, 6 mmol), diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) in N-methyl pyrrolidine (100 ml) overnight.
  • a linker group such as the t-butyl dimethyl silyl ether of 4-(methyl hydroxy-phenyl) acetic acid ( 1.3 g, 6 mmol), diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) in N-methyl pyrrolidine (100 ml) overnight.
  • the reaction is filtered through a glass frit under aspirator pressure and is washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 100 ml), then air dried.
  • the beads are then resuspended in THF (100 ml), and a desilylating agent such as tetrabutyl ammonium fluoride(6 ml, 1.0 M solution, 6 mmol, Aldrich) / ammonium acetate (0.92 g, 12 mmol) is used to deprotect the silyl ether producing the desired benzyl alcohol derivative.
  • a desilylating agent such as tetrabutyl ammonium fluoride(6 ml, 1.0 M solution, 6 mmol, Aldrich) / ammonium acetate (0.92 g, 12 mmol) is used to deprotect the silyl ether producing the desired benzyl alcohol derivative.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried.
  • the beads ( 5 g), as prepared in Procedure D, are then suspended in N- methyl pyrrolidine (100 ml) and is then reacted with a monomer such as FMOC-L- alanine (1.9 g, 6 eq., 6 mmol).
  • a monomer such as FMOC-L- alanine (1.9 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • the lysine derivatized beads (5.0 g), as described in Procedure A, are suspended in N-methyl pyrrolidine (100 ml), then a fluorophore such as 1-pyrene butyric acid (0.173 g, 0.6 eq., 0.6 mmol), and a doping agent such as butyric acid (0.48 g, 0.49 ml, 5.4 eq., 5.4 mmol) in 1:9 stoichiometry, and diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) are added, and the reaction is agitated for 3 hours.
  • a fluorophore such as 1-pyrene butyric acid (0.173 g, 0.6 eq., 0.6 mmol)
  • a doping agent such as butyric acid (0.48 g, 0.49 ml, 5.4 eq., 5.4 mmol) in 1:9 stoichiometry,
  • Th e reaction is filtered through a glass frit under aspirator pressure, washed with DMH ( 5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • the beads are then agitated in 25% TFA/ CH 2 CI 2 (100 ml) for 2 h removing the Boc protective group.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried.
  • the beads are then reacted with a linker group such as the t-butyl dimethyl silyl ether of 4-(methyl hydroxy-phenyl) acetic acid ( 1.3 g, 6 mmol), diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) in N-methyl pyrrolidine (100 ml) overnight.
  • a linker group such as the t-butyl dimethyl silyl ether of 4-(methyl hydroxy-phenyl) acetic acid ( 1.3 g, 6 mmol), diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) in N-methyl pyrrolidine (100 ml) overnight.
  • the reaction is filtered through a glass frit under aspirator pressure and is washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 100 ml), then air dried.
  • the beads are then resuspended in THF (100 ml), and a desilylating agent such as tetrabutyl ammonium fluoride(6 ml, 1.0 M solution, 6 mmol, Aldrich) / ammonium acetate (0.92 g, 12 mmol) is used to deprotect the silyl ether producing the desired benzyl alcohol derivative.
  • a desilylating agent such as tetrabutyl ammonium fluoride(6 ml, 1.0 M solution, 6 mmol, Aldrich) / ammonium acetate (0.92 g, 12 mmol) is used to deprotect the silyl ether producing the desired benzyl alcohol derivative.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried.
  • the beads ( 5 g), as prepared in Procedure F, are then suspended in N- methyl pyrrolidine (100 ml) and is then reacted with a monomer such as FMOC-L- phenylalanine (2.3 g, 6 eq., 6 mmol).
  • a monomer such as FMOC-L- phenylalanine (2.3 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • Pool H1 is reacted with a monomer such as FMOC-L-glyine (1.8 g, 6 eq., 6 mmol).
  • a monomer such as FMOC-L-glyine (1.8 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • Pool H2 is reacted with a monomer such as FMOC-L-alanine (1.9 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • Procedure J The beads obtained from Procedure I are then combined and sorted by flow cytometry into different sublibraries differentiated by the differences in intensity of fluorescence. Sublibrary components have the same first amino acid of the tripeptide.
  • Sublibrary J1 consists of Gly-X-X or Gly-Gly-Gly, Gly-Gly-Ala, Gly-Gly-Phe, Gly- Ala- Gly, Gly-Ala-Ala, Gly-Ala-Phe, Gly-Phe-Gly, Gly-Phe-Ala, Gly-Phe-Phe.
  • Sublibrary J2 consists of Ala-X-X or Ala-Gly-Gly, Ala-Gly-Ala, Ala-Gly-Phe, Ala- Ala- Gly, Ala-Ala- Ala, Ala-Ala-Phe, Ala-Phe-Gly, Ala-Phe-Ala, Ala-Phe-Phe
  • Sublibrary J3 consists of Phe-X-X or Phe-Gly-Gly, Phe-Gly-Ala, Phe-Gly-Phe, Phe- Ala- Gly, Phe-Ala-Ala, Phe-Ala-Phe, Phe-Phe-Gly, Phe-Phe-Ala, Phe-Phe-Phe-Phe
  • Pool M2 is reacted with a monomer such as FMOC-L-alanine (1.9 g, 6 eq., 6 mmol).
  • a monomer such as FMOC-L-alanine (1.9 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • Pool M3 is reacted with a monomer such as FMOC-L-phenylalanine (2.3 g, 6 eq., 6 mmol).
  • Diisopropyl carbodiimide (0.76 g, 0.94 ml, 6 mmol) is added, and the reaction is agitated for 3 hours.
  • the reaction is filtered through a glass frit under aspirator pressure, washed with DMF (5 x 30 ml), then CH 2 CI 2 (5 x 30 ml), then air dried. This procedure is repeated until the reaction is complete by the Kaiser ninhydrin test.
  • the beads obtained from Procedure M are then combined and sorted by flow cytometry into different sublibraries differentiated by the differences in intensity of fluorescence.
  • Sublibrary components have the same second amino acid of the tripeptide.
  • Sublibrary N1 consists of X-Gly-X or Gly-Gly-Gly, Gly-Gly- Ala, Gly-Gly-Phe, Ala- Gly-Gly, Ala-Gly-Ala, Ala-Gly-Phe, Phe-Gly-Gly, Phe-Gly-Ala, Phe-Gly- Phe
  • Sublibrary N2 consists of X-Ala-X or Gly-Ala-Gly, Gly- Ala- Ala, Gly-Ala-Phe, Ala- Ala- Gly, Ala- Ala- Ala, Ala-Ala-Phe, Phe-Ala-Gly, Phe-Ala-Ala, Phe-Ala- Phe
  • Sublibrary N3 consists of X-Phe-X or Gly-Phe-Gly, Gly-Phe-Ala, Gly-Phe-Phe, Ala-Phe- Gly, Ala-Phe-Ala, Ala-Ala-Phe, Phe-Phe-Gly, Phe-Phe-Ala, Phe- Phe-Phe-Phe
  • the beads obtained from Procedure P are then combined and sorted by flow cytometry into different pools differentiated by the differences in intensity of fluorescence.
  • the pools of beads obtained from Procedure Q are already sorted into sublibraries in which the third amino acid of each component is the same the same amino acid of the tripeptide.
  • Sublibrary Q1 consists of X-X-Gly or Gly-Gly-Gly, Gly-Ala-Gly, Gly-Phe-Gly,
  • Sublibrary Q2 consists of X-X-Ala or Gly-Gly- Ala, Gly- Ala- Ala, Gly-Phe- Ala,
  • Ala-Gly- Ala Ala- Ala- Ala, Ala-Phe-Ala, Phe-Gly-Ala, Phe-Ala-Ala, Phe-
  • Sublibrary Q3 consists of X-X-Phe or Gly-Gly-Phe, Gly-Ala-Phe, Gly-Phe-Phe,
  • Example 1 The methods of Example 1 are used except that the doping reagent is replaced by a second fluorophore such as perylene butyric acid.
  • a second fluorophore such as perylene butyric acid.

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Abstract

Sont décrits un procédé de préparation de bibliothèques combinatoires ainsi que les bibliothèques combinatoires ainsi obtenues. Est également décrit un procédé pour identifier, au moyen de la cytométrie de flux, des composés présentant des caractéristiques désirées dans une bibliothèque combinatoire ou un ensemble de bibliothèques combinatoires. Est également décrit un procédé pour coder des bibliothèques combinatoires à l'aide de billes à marqueurs fluorophores.
PCT/US1995/006392 1994-05-23 1995-05-23 Bibliotheques combinatoires codees Ceased WO1995032425A1 (fr)

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JP7530459A JPH10500951A (ja) 1994-05-23 1995-05-23 コードされた組合せライブラリー
US08/537,752 US6210900B1 (en) 1995-05-23 1995-05-23 Method of encoding a series of combinatorial libraries and developing structure activity relationships
EP95920576A EP0763202A4 (fr) 1994-05-23 1995-05-23 Bibliotheques combinatoires codees

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US24779394A 1994-05-23 1994-05-23
US26733394A 1994-06-28 1994-06-28
US38254295A 1995-02-01 1995-02-01
US41043695A 1995-03-23 1995-03-23
US08/267,333 1995-03-23
US08/382,542 1995-03-23
US08/410,436 1995-03-23
US08/247,793 1995-03-23

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Cited By (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5866341A (en) * 1996-04-03 1999-02-02 Chugai Pharmaceutical Co., Ltd. Compositions and methods for screening drug libraries
WO1999006839A1 (fr) * 1997-08-01 1999-02-11 Novalon Pharmaceutical Corporation Procede d'identification et de developpement de chefs de file de medicaments
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WO1999064867A1 (fr) * 1997-12-04 1999-12-16 Amersham Pharmacia Biotech Uk Limited Procede pour dosages multiples
US6054047A (en) * 1998-03-27 2000-04-25 Synsorb Biotech, Inc. Apparatus for screening compound libraries
WO2000032542A1 (fr) * 1998-11-30 2000-06-08 The University Of Queensland Supports pour banques combinatoires de composes
WO2000061281A1 (fr) * 1999-04-09 2000-10-19 Glaxo Group Limited Systeme de codage pour banques de produits chimiques en phase solide
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US6506342B1 (en) * 1997-06-19 2003-01-14 Robert D. Frankel Tracking apparatus and method for use with combinatorial synthesis processes
US6602685B1 (en) 1998-08-17 2003-08-05 Phylos, Inc. Identification of compound-protein interactions using libraries of protein-nucleic acid fusion molecules
US6607921B1 (en) 1998-03-27 2003-08-19 Ole Hindsgaul Methods for screening compound libraries
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US6878557B1 (en) 1995-01-20 2005-04-12 Arqule, Inc. Logically ordered arrays of compounds and methods of making and using the same
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US7022479B2 (en) 2000-01-24 2006-04-04 Compound Therapeutics, Inc. Sensitive, multiplexed diagnostic assays for protein analysis
US7034110B2 (en) 1994-01-05 2006-04-25 Arqule, Inc. Method of identifying chemical compounds having selected properties for a particular application
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WO2017129998A1 (fr) 2016-01-29 2017-08-03 Heptares Therapeutics Limited Protéines g
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WO2020049130A1 (fr) 2018-09-05 2020-03-12 Valerie Nicholas Carl Kristoffer Procédés
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Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20020026421A (ko) * 1999-04-15 2002-04-10 버츄얼 어레이즈 인코포레이티드 코딩 위치에 표시를 갖는 조합적인 화학 라이브러리지지체 및 이의 사용 방법
CA2398700A1 (fr) * 2000-02-07 2001-08-16 Kyowa Medex Co., Ltd. Procede de detection d'une substance

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
NATURE, Volume 354, issued 07 November 1991, LAM et al., "A New Type of Synthetic Peptide Library for Identifying Ligand-Binding Activity", pages 82-84. *
PROC. NATL. ACAD. SCI. U.S.A., Volume 90, issued December 1993, OHLMEYER et al., "Complex Synthetic Chemical Libraries Indexed with Molecular Tags", pages 10922-10926. *
PROC. NATL. ACAD. SCI. U.S.A., Volume 90, issued November 1993, NEEDELS et al., "Generation and Screening of an Oligonucleotide-Encoded Synthetic Peptide Library", pages 10700-10704. *
See also references of EP0763202A4 *

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US10393759B2 (en) 2011-04-12 2019-08-27 Quanterix Corporation Methods of determining a treatment protocol for and/or a prognosis of a patient's recovery from a brain injury
US11275092B2 (en) 2011-04-12 2022-03-15 Quanterix Corporation Methods of determining a treatment protocol for and/or a prognosis of a patient's recovery from a brain injury
US12571803B2 (en) 2011-04-12 2026-03-10 Quanterix Corporation Methods of producing bodily fluid samples containing an analytically quantified amount of phosphorylated tau protein
US12540950B2 (en) 2011-04-12 2026-02-03 Quanterix Corporation Methods of assaying phosphorylated tau protein in a sample
US9932626B2 (en) 2013-01-15 2018-04-03 Quanterix Corporation Detection of DNA or RNA using single molecule arrays and other techniques
US10640814B2 (en) 2013-01-15 2020-05-05 Quanterix Corporation Detection of DNA or RNA using single molecule arrays and other techniques
WO2016116760A2 (fr) 2015-01-21 2016-07-28 Cancer Research Technology Limited Inhibiteurs
US11919959B2 (en) 2015-01-21 2024-03-05 Cancer Research Technology Limited Inhibitors of the interaction between CLEC14A and multimerin-2 for inhibition of angiogenesis
US10808031B2 (en) 2015-01-21 2020-10-20 Cancer Research Technology Limited Inhibitors of the interaction between CLEC14A and multimerin-2 for inhibition of angiogenesis
WO2017129998A1 (fr) 2016-01-29 2017-08-03 Heptares Therapeutics Limited Protéines g
EP4516806A2 (fr) 2016-01-29 2025-03-05 Nxera Pharma UK Limited Proteines g
US10738287B2 (en) 2016-01-29 2020-08-11 Heptares Therapeutics Limited G proteins
US11339383B2 (en) 2016-01-29 2022-05-24 Heptares Therapeutics Limited G proteins
WO2020049130A1 (fr) 2018-09-05 2020-03-12 Valerie Nicholas Carl Kristoffer Procédés

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