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WO1995013538A1 - Procede d'obtention et de criblage de bibliotheques de composes chimiques complexes - Google Patents

Procede d'obtention et de criblage de bibliotheques de composes chimiques complexes Download PDF

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Publication number
WO1995013538A1
WO1995013538A1 PCT/US1994/012956 US9412956W WO9513538A1 WO 1995013538 A1 WO1995013538 A1 WO 1995013538A1 US 9412956 W US9412956 W US 9412956W WO 9513538 A1 WO9513538 A1 WO 9513538A1
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Prior art keywords
molecules
adduct
core molecule
screening
core
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PCT/US1994/012956
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English (en)
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Robert L. Saul
Eric K. Pham
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Operon Technologies, Inc.
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Publication of WO1995013538A1 publication Critical patent/WO1995013538A1/fr

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Classifications

    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
    • C07C41/01—Preparation of ethers
    • C07C41/16—Preparation of ethers by reaction of esters of mineral or organic acids with hydroxy or O-metal groups
    • 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
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/08—Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
    • 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
    • B01J2219/0068—Means for controlling the apparatus of the process
    • B01J2219/00702—Processes involving means for analysing and characterising the products
    • 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

Definitions

  • the field of this invention concerns the preparation of complex mixtures of organic compounds, identification of activity, and identification of individual compounds having the activity.
  • Synthetic organic compounds play an extraordinary role in the well being of modern society. Synthetic organic compounds find application as therapeutics, as pesticides, as additives in a wide variety of context, as dyes, as well as in many other environments. Over the years numerous synthetic organic compounds have been produced, particularly those compounds, where there has been some ab initio reason for believing that the compound would have an activity of interest. This belief could be predicated upon an activity of a natural occurring compound, some insight into the nature of the target, or the like. The breadth of compounds that have been produced is quite staggering in number, but the variety of opportunities in producing organic compounds, due to the versatility of carbon and the numerous groups that can be used to functionaiize carbon atoms, leaves a vast unexplored area.
  • Combinatorial chemistry is a very powerful strategy. Combinatorial libraries make large additions to the chemical databases, typically used in the pharmaceutical industry; combinatorial chemistry effectively combines all the laborious tasks in traditional drug discovery, e.g. collection and testing of natural product samples, isolation and purification of active ingredients, structure determination and synthesis, into one rapid series of steps; since combinatorial chemistry creates such a large pool of potential active compounds, the candidate compounds are often more specific than those found in nature or synthesized by traditional means; small organic compounds make up the vast majority of drugs and other physiologically active compounds, and large numbers of small organic molecules can be made over short periods of time.
  • the methodology requires an accessible means for determining the nature of a product which is active as to a particular property or characteristic. The requirement for identification of a product which is found to be active has severely limited the approaches used to produce large libraries.
  • oligomers such as oligopeptides and oligonucleotides. These approaches have involved a wide variety of methods for producing the oligomers and determining their structure, as well as screening the oligomers for activity. However, despite the very substantial efforts involved and the numerous protocols for their preparation and screening, there have been no reported significant successes in therapeutically useful drug development.
  • One of the problems is that the oligomers are relatively flexible and it is generally believed that fairly rigid molecules will be required for binding, where a target molecule is involved, to achieve the end purpose.
  • linear biopolymers such as oligonucleotides, peptides and peptidomimetics suffer from poor oral bioavailability, being too large to be absorbed and too readily degraded physiologically. Efforts have therefore been directed to synthetic techniques for synthesizing small, organic molecules. Bunin and Ell an, J. Am . Chem . Soc . 114, 10997 (1992) , reported a combinatorial approach to the synthesis of diazepams. By using a few steps and a relatively limited repertoire, Ellman was able to synthesize a library of diazepam compounds. This approach is fairly restricted in the number and type of compounds which can be produced, since the products must be produced one at a time.
  • Patents of interest include U.S. patent nos. 5,182,366; 5,270,170; and 5,252,296.
  • PCT applications of interest include
  • a method for producing a library of related molecules employing a polyfunctional core molecule and a plurality of adduct molecules capable of reacting with the core molecule functionalities.
  • a plurality of non- overlapping adduct subset units are defined to be used to define the adduct reaction set.
  • As a first or intermediate step in each of a series of vessels one of overlapping different sets of adduct molecules comprised of less than the total number of subset units are added to the core molecules under reaction conditions, where usually the reaction rate in each vessel of each of the adduct molecules is substantially normalized. The content of each vessel is then screened for activity as to one or more utilities.
  • a second series of reactions may be carried out using the core molecules and a set of adduct molecules having a reduced number per vessel of adduct molecules coming from within the set which was active.
  • This set may be selected to ensure that all the molecules produced in the active set are produced or, by comparing the results of the overlapping sets, one or more of the adduct molecules may be excluded from the set as not being involved in an active product.
  • a screening program is provided to rapidly narrow a large number of candidate molecules to a relatively small number of candidate molecules, which may then be characterized by physical or other means as to the identity of the active compound(s).
  • complex mixtures which are non-polymeric synthetic organic molecules are produced, where a plurality of vessels are employed, each vessel containing the same polyfunctional core molecule.
  • the polyfunctional core molecule may have the same functionalities [poly(mono)functional] or different functionalities having similar reactivity to the adducts [poly(multi)functional] .
  • adducts a set of reactants capable of reacting with the functionalities present on the core molecule.
  • Non- overlapping subsets of adducts are chosen to define overlapping different sets of adducts.
  • the sets are selected to provide all possible combinations of products resulting from reactions with the members of all of the sets.
  • the reaction is allowed to proceed in each of the vessels to at least substantial completion.
  • the conditions of the reaction including the concentrations of each of the adducts, are selected so as to substantially normalize the rate of reaction of each of the adducts with the core molecule functionality and to minimize undesired side reactions.
  • all or aliquots of a reaction mixture from a single vessel may then be screened for activity as to a particular purpose. Any activity may be considered or a minimal threshold activity may be selected for a particular mixture as defining activity.
  • a second series of reactions may be carried out.
  • a plurality of vessels is employed, where the core molecule is present in each vessel, and individual sets of adducts are added to each vessel, where the sets have fewer adducts than the previous set, and may provide less than all of the compounds which were produced in the previous reaction vessel up to all of the compounds.
  • this may be as a result of analysis of activity of the various mixtures, demonstrating that certain combinations will not be active or allowing for the absence of certain combinations, where lack of activity in a vessel would indicate that the compound(s) which was not produced provided the activity in the previous vessel.
  • This process may be repeated as many times as necessary to obtain a single product or a mixture of products which can be analyzed, so as to identify the individual products in the mixture, where the individual synthesis of all the products is not onerous.
  • the number of adduct molecules will usually be at least 6, more usually at least 8, generally at least about 12, usually not exceeding 200, more usually not exceeding 100.
  • the step or stage comprising a plurality of adducts, where sets of adducts are used in different reaction vessels with the same core molecules may be the first step or an intermediate step. While one may carry out a single reaction in a single vessel with a set of all the adducts, and then screen the entire mixture for activity, generally this will not be done. More usually, one will begin as a first step using a plurality of vessels, at least two, more usually at least three, preferably at least about five, and usually not more than 300, more usually not more than 200, and preferably not more than 100. The number of vessels will be dictated by the number of functionalities on the core molecule, the number of adducts, the desired complexity of the mixtures, the sensitivity of the assay(s) for screening activity, the amounts of products needed, and the like.
  • the system will provide for the preparation of at least about 100 compounds, more usually at least about 200 compounds, preferably at least about 500 compounds, frequently at least about 1,000 compounds, usually fewer than about 5 x 10 6 compounds, more usually fewer than about 1 x 10° compounds. While for the most part it may be assumed that the primary products will be the per-substituted core molecule, one may anticipate that in many situations, there will be small but identifiable amounts of core molecules which have reacted at fewer than the total number of functionalities present.
  • a "primary" list of adduct molecules is made. This list defines the variety of products which may be achieved in relation to the core molecule. As a convenient tool, this list may be divided into non-overlapping subset units, where the number of units is equal to or greater than the number of reactive functionalities on the core molecule. These units are then combined to provide different overlapping sets of adducts for reaction in different vessels, so that the total number of possible products from the adducts is obtained. Each vessel will have a different mixture of products, having a proportion of products which overlap the products in other vessels.
  • screening is improved in that fewer than the total number of products is present in each vessel, allowing for fewer iterative steps to define active compounds, with less chance for interactions between compounds confusing the results of the assays.
  • activity in a particular vessel one can then repeat the process, using the list of adduct molecules used in that particular vessel to define the total number of adduct molecules which are to be distributed among the plurality of vessels in accordance with the previously described strategy in the next step.
  • the total number of products produced by the subject method is indicated by the following:
  • R'-X + Z-(Y) V —> Zf-W-R 1 ),. where i l to n.
  • the products all have the same core moiety "Z,” but have random covalently bonded adduct moieties, where the R groups attached to the core moiety may be the same or different.
  • X is a reactive functionality, which reacts with Y to form a covalent bond.
  • W is the linking group formed from the reaction of X and Y. Illustrative of the reaction situation would be a core molecule with four reactive functionalities.
  • the screening strategy should efficiently narrow the number of adduct molecules used at each step, while ensuring that each of the possible products produced by the original adduct set is formed in at least one of the subsets.
  • the primary adduct set, S is divided into "k" non-overlapping partial subsets, p j , with equal numbers of adducts (> 1 adduct per subset) , where a subscript "i” denotes an integer of from 1 to k (k being greater than the number of reactive functionalities on the core molecule) .
  • Secondary subsets s 2 ⁇ j are then prepared by making each possible combination or union set of "v" partial subsets (v is the number of reactive functionalities on the core molecule and the subscript j is an integer from 1 to m, where m is the number of subsets) .
  • the secondary subsets are then used to synthesize secondary product mixtures for screening.
  • a trifunctional asymmetrical core molecule is reacted with a primary adduct set of 12 different adduct molecules
  • the original adduct set of 12 adduct molecules (termed A,B,C,D,E,F,G,H, I,J,K,L) is divided into a number of non-overlapping partial subset units.
  • the number of partial subset units, k is selected to be greater than the number of reactive functionalities on the core molecule.
  • the original set of 12 adduct types has been reduced to four subsets of 9 adduct types.
  • the original set of 12 adduct types has been reduced to 20 subsets of 6 adduct types.
  • 20 adduct reagent mixtures would be prepared and separately reacted with the core molecule to produce 20 different product mixtures for screening.
  • a series of 20 smaller tertiary adduct subsets could be defined to continue the process.
  • one iteration of the screening process decreases the number of adducts per subset by the factor 1/2, and the number of products per mixture by 1/8. These factors would narrow an adduct list in significantly fewer steps. However, a larger number of product mixtures must be screened to achieve this narrowing of the adduct list in fewer iterations.
  • the screening assays are automated and a 96-well microliter plate is employed, it is efficient to produce as many as 96 different product mixtures or more at each step.
  • the Table below provides some preferred values for the number of partial subsets, the number of secondary product mixtures and the reduction in the number of adduct types for each iteration when core molecules with two, three, four and five reactive functionalities are used.
  • the reduction factor (v/k) is the amount by which the number of adduct types are reduced after each step of screening.
  • Preferred reduction factors are those having reciprocals which are integers. Such reduction factors can be repeatedly and evenly applied to initial adduct sets.
  • the vessels employed may be any convenient vessels, which provide for the desired volume.
  • the core molecule will normally be present in an amount of from about 0.1 millimole to 10 illimole.
  • each adduct molecule will be present in at least about 0.5 millimole, usually at least about 2 millimoles and can be up to 100 millimoles or more in each mixture, usually not exceeding about 50 millimoles, more usually not exceeding about 10 millimole in each mixture.
  • the amount of individual compounds which will be produced will be related to the minimal activity of interest, sensitivity of the assay, ease and economics of synthesis, and the like.
  • Reaction volumes will usually be at least about 1 ml, more usually at least about 5 ml, and may be 0.5 L or more, usually not exceeding about 100 ml.
  • concentration of the core molecule will generally be in the range of about 1 mM to 10M, more usually about 10 mM to 1 M.
  • the core molecule will be in a mol ratio of about 0.02 to 1 to each of the adduct molecules, depending on the relative rate of each of the adduct molecules, the desired rate of reaction, the volume of the reaction mixture, and the like.
  • Solvents will be chosen in accordance with the nature of the reaction, where the solvents may be polar or non-polar organic or inorganic, combinations thereof, single or mixtures of solvents, and the like.
  • Illustrative solvents include dimethylformamide, DMSO, ethyl ether, propanol, acetonitrile, toluene, anisole, acetone, and the like.
  • the core molecule and adducts will be combined, either neat or in the presence of a solvent, to carry out the reaction to form the mixture. Temperatures will depend upon the particular reaction, generally ranging from about -10 to 100°C.
  • the core molecule when present initially, will generally be at a concentration in the range of about 1 mM to 1 M, more usually in the range of about 10 mM to 100 mM.
  • concentrations of the reactants may vary depending upon the manner and order of addition.
  • the concentrations will be selected, so that there will be a reasonable opportunity to prepare all possible per- substituted compounds, where the difference in rate will not result in substantial exhaustion of one or more adducts during a period where there has been little or no reaction of one or more other adducts.
  • the relative rates of reaction of the individual adducts can be readily determined by setting up a pairwise experiment in which two adducts are reacted with the core molecule.
  • the relative rates of reaction of the adduct of interest is directly related to the factor which affords equimolar yields of the two aforementioned adducts. In this way, one can readily define the various ratios to be used in each of the vessels for production of a particular composition mixture. Desirably the yield range should not be greater than about 2 to 10, preferably not greater than about 2 to 5.
  • Another method for normalization is to choose one adduct reagent to be a "normalization standard," and for each of the other adduct reagents, prepare an equimolar binary mixture of that reagent and the normalization standard. Each binary adduct mixture is then reacted with the core molecule and the "relative reactivity" of each adduct relative to the normalization standard is readily determined from the product yields.
  • the core molecule is added to the preformed mixture of adducts, the core molecule and adducts are added substantially simultaneously or the adducts are added to the core molecule at the same or different rates and at the same or different concentrations, to substantially maintain the normalized rate of reaction of the various adducts.
  • the adduct molecules will usually be employed in greater than stoichiometric amounts, (the number of moles being equivalent to the proportion of reactive functionalities which the adduct will react with based on the number of adduct molecules and the number of different adduct reagents) , generally in at least 2 fold molar excess, usually at least about 5 fold molar excess, and may be about 75 fold molar excess or more, generally less than about 50 fold molar excess.
  • the amounts of each of the adduct molecules will be relatively similar.
  • the core molecule may be free in solution or be bound to a solid support, where the solid support may be resins, e.g. Merrifield or modified versions thereof, silica-based support, e.g. aminopropyl silica (ref. J. Am. Chem. Soc. 116:1135 [1994]), solid surfaces such as magnetic particles, porous glass beads, polyethylene pins, etc.
  • the solid support may be resins, e.g. Merrifield or modified versions thereof, silica-based support, e.g. aminopropyl silica (ref. J. Am. Chem. Soc. 116:1135 [1994]), solid surfaces such as magnetic particles, porous glass beads, polyethylene pins, etc.
  • silica-based support e.g. aminopropyl silica (ref. J. Am. Chem. Soc. 116:1135 [1994])
  • solid surfaces such as magnetic particles, porous glass beads, polyethylene pins, etc.
  • a wide variety of molecules can be cleaved preferentially, e.g., thiophenyl ethers or silyl compounds which may be cleaved by mercuric trifluoroacetate or fluorides (tetrabutylam onium fluoride) , nitrobenzyl ethers, which can be cleaved by photolysis, etc.
  • reaction will be carried out with agitation, so as to ensure a substantially uniform mixture.
  • Agitation can be achieved by rocking, stirring, shaking, or other convenient means.
  • the adducts may have the same or different functionality, so long as the required normalization can be achieved.
  • the functionalities on the adduct molecules may include such reactive groups as halogen, usually other than fluorine, where the halogen may be used in a Grignard (RMgX) or Reformatsky (RZnX) reaction, oxy, oxo, ketone or aldehyde, carboxylic acids and derivatives thereof such as acid halides, esters and anhydrides, amino, hydrazino, ylides, cyanate, isocyanate, isothiocyanate, olefinic or acetylenic unsaturation, small alicyclic or heterocyclic rings, such as cyclopropyl, oxiranyl, and aziridinyl, phosphoric acids, such as phosphates, phosphinates, phosphinamides, phosphoryl hal
  • phosphoric acids such as phosphates, phosphinates, pho
  • Functionalities which may be produced include esters and amides, from both organic and inorganic acids, ethers, both oxy and thio, imines, hydrazones, chiral epoxides from the asymmetric oxidation of alkenes with an enantioselective catalyst (J. Am. Chem. Soc. 116:6937-8 [1994]), etc.
  • the adduct molecules will be selected to substantially reduce or eliminate unimolecular or bimolecular side reactions, such as elimination, condensation, addition, and the like. For the most part, with active halides, elimination will be the major side reaction under basic or acidic conditions. Therefore, substitution at the a- or /3-position will usually be limited to the presence of at least one hydrogen atom for aliphatic molecules and heterofunctionalities or other functionalities, which might aid elimination will be avoided at these positions. Ester and carbonyl groups will usually be avoided in basic media. The reactions of the common functionalities are well known and obvious side reactions can be avoided.
  • the core molecule may be aliphatic, alicyclic, aromatic or heterocyclic, where the heteroatoms will usually be nitrogen, oxygen and sulfur, although other heteroatoms, such as phosphorous, metal atoms, boron, etc. may be present.
  • metallocenes may serve as the core.
  • the active functionalities may be symmetrically situated, so as to have the same reactivity or may be asymmetrically situated, so as to have differing activity.
  • Compounds of interest include sugars, such as mono- and disaccharides, polyfunctionalized aromatic compounds, where the aromatic core may be carbocyclic or heterocyclic, such as benzene, naphthalene, biphenyl, pyridine, etc. , may be alicyclic where the rings may be mono- or bicyclic or higher order, such as cyclohexane, cyclooctane, adamantane, bicyclo-heptane; acyclic organic compounds, such as ethylenediamine tetracetic acid, nitrolotriacetic acid, tris-hydroxyethyla ine, 2,2,2-tri- thiolmethyl-1-methoxyethane, etc.
  • active functionalities on the core molecule will preferably be separated by a sufficient distance to minimize steric hindrance or other interactions which would substantially change the reactivity of an active functionality with the adduct reagents, either before the reaction of one of the active functionalities or after reaction of one of the reactive functionalities with an adduct reagent. While not universally true, two active functionalities will be separated by at least about 3 atoms and be bonded to an atom, usually carbon, which when saturated will usually be bonded to at least one hydrogen atom. Steric effects may be diagnosed by running reactions with a core molecule and a single adduct type and verifying that the main product is per-substituted.
  • the core molecules may be symmetrical or asymmetrical
  • the symmetrical core molecules yield fewer products, but the products are easier to separate and identify, as compared to asymmetrical molecules.
  • 1,2,6-hexane and 1,3,5- trihydroxybenzene the latter compound has both a C 3 and C 2 axis of symmetry.
  • the trigonal symmetry in 1,3,5- trihydroxybenzene reduces this to 35 as the number of distinct trisubstituted products is given by the formula, (n 3 + 3n 2 + 2n)/6.
  • adduct molecules may be difunctional, where the adduct molecule can react with two functionalities on the core molecule to form a ring.
  • the core molecule will usually have two vicinal functionalities or be situated in spatial proximity, e.g. 1,8-disubstituted naphthalene, where the resulting rings will have from about 5 to 7 annular members.
  • Illustrative situations include dihalides with glycols, activated dicarboxylic acids with diamines, diisocyanates with diamines, etc.
  • the linking group formed by the reaction between the core molecule functionality and the adduct functionality may take many forms, including carbon-carbon, carbon- oxygen, carbon-nitrogen and carbon-sulfur bonds.
  • the reactions may be addition, substitution, elimination, condensation, free radical, or other reaction, as appropriate. Reactions which may be involved include esterification, amidification, etherification, addition to unsaturation, Claisen condensation, metal catalyzed coupling reactions, asymmetric epoxidation or hydroxylation, etc.
  • For carbon-carbon bond formation one may use the Grignard reagent with an oxo or non-oxo- carbonyl functionality or the oxo functionality or an active halide with an organolithium compound.
  • Suitable reactions may be found in March, "Advanced Organic Chemistry” , 4th ed. , Wiley-Interscience, NY, 1992, incorporated herein by reference.
  • pages 417-424 relate to amide formation, page 903 to urea formation, pages 896-898 to imine formation, pages 386-387 to ether formation and pages 920-930 to Grignard reactions.
  • Reaction conditions will be optimized to provide for high yields of completely reacted core molecules, where all of the reactive functionalities have reacted with the adduct molecules. Conditions can be chosen to drive the reaction to completion, such as large excesses of the adduct molecules, elevated temperatures and pressures, long reaction times, where feasible, segregation of product from the main reaction mixture, catalysts, or the like.
  • Segregation can be achieved where the final product has a different solubility from the core molecule and intermediate products, so that the final product may be taken up in a selective solvent.
  • Functionalization of certain functionalities to enhance reactivity may be employed. With carboxylic acids, active esters may be formed, e.g. pentafluorophenyl or o-nitrophenyl esters, or carbodiimides may be added. Lithium derivatives may be prepared from amides or amines. Metal ions may be added to enhance substitution reactions. The creation of stereogenic centers via asymmetric induction with chiral catalysts may also be used.
  • chiral secondary alcohols may be prepared from the enantioselective reduction of ketones, while highly enantioselective formation of epoxides can result from the oxidation of simple olefins in the presence of a chiral manganese catalyst.
  • the reaction mixture may be subjected to a variety of treatments. Any particular treatment will depend upon the nature of the products, the purpose for which the product is to be screened, the solvents used, and the like. Any remaining adduct molecules may be removed by any convenient means. In some situations where an organic solvent is used, the solvent may be removed and the resulting product dispersed in an aqueous medium, an aqueous organic medium, or organic medium e.g., DMSO, DMF, ethanol, etc. Where the product has been produced bound to a surface, the product may be released from the surface as described previously, based upon the labile linking group.
  • protective groups may have been employed with functionalities present on the adducts.
  • the protective groups may be removed in accordance with conventional ways, depending upon the nature of the protective group, as well as the nature of the products. For example, amino groups may be protected with FMOC, where the FMOC groups may be removed with piperidine in DMF.
  • Various techniques may be employed to separate the products from the excess adduct molecules. Such techniques as distillation, chro atography, e.g. ion exchange chromatography, solvent extraction, base or acid extraction, or the like, may be employed in accordance with the nature of the products and adducts. If desired, the product mixture may be fractionated in accordance with a particular characteristic, to give more information about the nature of an active product.
  • the product mixture is then screened for activity.
  • the product mixture may be screened for one or more activities, depending upon the nature of the product.
  • One screening may be for binding affinity.
  • the subject compositions may be screened for their ability to bind specifically with reasonable specificity to a target molecule.
  • the subject compositions may be screened as enzyme inhibitors, as molecules for directing another molecule to a specific site, e.g. a physiological site, such as a blood protein, a surface membrane protein, specific organs, plant parts, pathogens, and the like.
  • a specific site e.g. a physiological site, such as a blood protein, a surface membrane protein, specific organs, plant parts, pathogens, and the like.
  • the subject compounds will be evaluated for activity, as cytotoxic agents, for inducing a signal across a surface membrane protein, as a competitor to a natural ligand, or the like.
  • the subject compounds may also find use in diagnostics, where they may serve as reagents for competition with an analyte, binding to a ligand of interest, as labels, enzyme substrates, and the like.
  • the subject compounds may also be evaluated for binding to nucleic acids, sugars, etc.
  • screening for determining binding affinity, a number of different techniques may be employed. One may pass the mixture through a column in which the target molecule is bound to a solid support. From such a column, weakly binding or non-binding compounds will be eluted.
  • an isocratic or gradient solvent with increasing polarity or increasing ionic strength so as to obtain different fractions of compounds which bind, depending upon the effect of the solvent on the binding affinity.
  • a fraction has one or few compounds, usually fewer than five, one may be able to analyze the compounds in the fraction to determine their structure. Even if a complete structure cannot be determined, information may be obtained so as to reduce the number of adducts required at the next stage.
  • cytotoxic or stimulating effect By combining the product mixture with a target unicellular microorganism in an appropriate medium, one can determine the rate of proliferation of the microorganism in the presence or absence of the product mixture. Stimulation or inhibition may be determined.
  • Other screens may include binding to mammalian cells, where the target may be associated with homing, transduction of the signal across the membrane, inhibition of T-cells, binding to MHC antigens, etc.
  • the subject compounds may be screened for a wide variety of applications as additives, for fuels, oils, hydraulic fluids, boilers, plastics, food, cosmetics, etc., as pesticides, stabilizers, antioxidants, etc.
  • the mixture will be employed in accordance with conventional methods for evaluating a particular performance.
  • the process is then repeated, where a plurality of vessels are employed, but the primary set of adduct molecules is now up to and including the total number used for a particular vessel in the previous stage, where the product mixture from the vessel is found to be active.
  • the number of vessels required and the adducts introduced in each vessel may be substantially reduced or expanded, as compared to the first stage, depending upon how few compounds are to be produced in each vessel. This iterative process may be repeated as many times as necessary, until one obtains a single compound or a mixture of compounds which can be individually analyzed.
  • the adduct reagents were primary alkyl bromides: bromopropane (A) ; 3-acetoxypropyl-l bromide (B) ; phenoxyethyl bromide (C) ; D-citronellyl bromide (D) ; and 3-cyanopropyl-l bromide (E) .
  • the reaction was carried out as described in Exaple 1. Mixed derivitizations were performed, by combining resorcinol pairing each of the alkyl bromides with A to determine the relative reactivity under the reaction conditions for each of the alkyl bromides normalized to the reactivity of A.
  • the weight ratios of the reactants in the reaction mixture were: Alkyl Bromide Relative Amount used Reactivity mmol
  • the conditions for the normalization were 1 mmol phloroglucinol, 15 mmol of KF/A1 2 0 3 15 mmol of propyl bromide and 15 mmol of the test bromide in DMF at 25°C.
  • the following table indicates the results of the rate normalization of 10 aliphatic bromides.

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  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Structural Engineering (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
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  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

Procédé de production de mélanges complexes de composés utilisant plusieurs cuves dans la première étape et la suivante. Chacune des cuves contient une molécule de base plurifonctionnelle dont les fonctionnalités entraînent des réactivités similaires, et des molécules additionnnelles présentant un taux de réaction similaire dans les conditions de la réaction. Chacune des cuves contient un jeu de molécules additionnnelles se recouvrant mais différentes de façon à fournir divers ensembles de produits. Lesdits produits subissent ensuite un criblage par activité au cours duquel un mélange réactif particulier est analysé en répétant le processus de division des molécules additionnelles dans des ensembles, se recouvrant mais différents, contenus dans différentes cuves, puis en criblant les cuves par activité. On peut de cette façon obtenir un composé unique ou un groupe de composés pouvant être analysés ou criblés de manière à définir le ou les composés présentant les activités indiquées.
PCT/US1994/012956 1993-11-12 1994-11-10 Procede d'obtention et de criblage de bibliotheques de composes chimiques complexes WO1995013538A1 (fr)

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WO1996040732A1 (fr) * 1995-06-07 1996-12-19 Zymogenetics, Inc. Banques combinatoires de composes non peptidiques
DE19609272A1 (de) * 1996-02-27 1997-08-28 Schering Ag Verfahren zur Parallelsynthese großer Zahlen von 2,6-disubstituierten Chinolinderivaten sowie 2,6-disubstituierte Chinolinderivate selbst
WO1998016536A1 (fr) * 1996-10-16 1998-04-23 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts Bibliotheque de saccarides
EP0739486A4 (fr) * 1994-01-13 1998-09-09 Univ Columbia Recepteurs de synthese, leurs bibliotheques et leurs utilisations
WO1998020020A3 (fr) * 1996-11-06 1998-10-22 Sequenom Inc Immobilisation haute densite d'acides nucleiques
WO1999064867A1 (fr) * 1997-12-04 1999-12-16 Amersham Pharmacia Biotech Uk Limited Procede pour dosages multiples
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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
WO2005044962A3 (fr) * 2003-10-31 2005-09-01 Chevron Oronite Co Preparation a haut rendement de compositions d'huiles lubrifiantes pour bibliotheques combinatoires
US6949633B1 (en) 1995-05-22 2005-09-27 Sequenom, Inc. Primers useful for sizing nucleic acids
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WO2014052174A1 (fr) * 2012-09-25 2014-04-03 Merck Sharp & Dohme Corp. Diversification de composé à l'aide d'une fonctionnalisation au stade tardif
US8999266B2 (en) 2000-10-30 2015-04-07 Agena Bioscience, Inc. Method and apparatus for delivery of submicroliter volumes onto a substrate
US9068953B2 (en) 2007-09-17 2015-06-30 Agena Bioscience, Inc. Integrated robotic sample transfer device

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Publication number Priority date Publication date Assignee Title
EP0739486A4 (fr) * 1994-01-13 1998-09-09 Univ Columbia Recepteurs de synthese, leurs bibliotheques et leurs utilisations
US7067326B2 (en) 1994-01-13 2006-06-27 The Trustees Of Columbia University In The City Of New York Synthetic receptors, libraries and uses thereof
US6797522B1 (en) 1994-01-13 2004-09-28 The Trustees Of Columbia University In The City Of New York Synthetic receptors
US6878557B1 (en) 1995-01-20 2005-04-12 Arqule, Inc. Logically ordered arrays of compounds and methods of making and using the same
US6949633B1 (en) 1995-05-22 2005-09-27 Sequenom, Inc. Primers useful for sizing nucleic acids
WO1996040732A1 (fr) * 1995-06-07 1996-12-19 Zymogenetics, Inc. Banques combinatoires de composes non peptidiques
DE19609272A1 (de) * 1996-02-27 1997-08-28 Schering Ag Verfahren zur Parallelsynthese großer Zahlen von 2,6-disubstituierten Chinolinderivaten sowie 2,6-disubstituierte Chinolinderivate selbst
DE19609272C2 (de) * 1996-02-27 2000-05-25 Schering Ag Verfahren zur Parallelsynthese großer Zahlen von 2,6-disubstituierten Chinolinderivaten
WO1998016536A1 (fr) * 1996-10-16 1998-04-23 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts Bibliotheque de saccarides
US7501251B2 (en) 1996-11-06 2009-03-10 Sequenom, Inc. DNA diagnostics based on mass spectrometry
EP1457496A1 (fr) * 1996-11-06 2004-09-15 Sequenom, Inc. Immobilisation haute densité d'acides nucléiques
WO1998020020A3 (fr) * 1996-11-06 1998-10-22 Sequenom Inc Immobilisation haute densite d'acides nucleiques
US7198893B1 (en) 1996-11-06 2007-04-03 Sequenom, Inc. DNA diagnostics based on mass spectrometry
US7132519B2 (en) 1996-12-10 2006-11-07 Sequenom, Inc. Releasable nonvolatile mass-label molecules
US7285422B1 (en) 1997-01-23 2007-10-23 Sequenom, Inc. Systems and methods for preparing and analyzing low volume analyte array elements
WO1999064867A1 (fr) * 1997-12-04 1999-12-16 Amersham Pharmacia Biotech Uk Limited Procede pour dosages multiples
US7550410B2 (en) 1999-05-05 2009-06-23 Foote Robert S Method and apparatus for combinatorial chemistry
US7179591B2 (en) 1999-05-05 2007-02-20 Robert S. Foote Method and apparatus for combinatorial chemistry
US8193336B2 (en) 1999-05-05 2012-06-05 Foote Robert S Method and apparatus for combinatorial chemistry
US9669376B2 (en) 2000-10-30 2017-06-06 Agena Bioscience, Inc. Method and apparatus for delivery of submicroliter volumes onto a substrate
US8999266B2 (en) 2000-10-30 2015-04-07 Agena Bioscience, Inc. Method and apparatus for delivery of submicroliter volumes onto a substrate
WO2005044962A3 (fr) * 2003-10-31 2005-09-01 Chevron Oronite Co Preparation a haut rendement de compositions d'huiles lubrifiantes pour bibliotheques combinatoires
WO2005045205A3 (fr) * 2003-10-31 2005-10-20 Chevron Oronite Co Bibliotheques combinatoires de compositions d'huiles lubrifiantes
US7579192B2 (en) * 2003-10-31 2009-08-25 Chevron Oronite Company Llc High throughput screening methods for lubricating oil compositions
US7468280B2 (en) 2003-10-31 2008-12-23 Chevron Oronite Company Llc High throughput preparation of lubricating oil compositions for combinatorial libraries
US7462490B2 (en) * 2003-10-31 2008-12-09 Chevron Oronite Company Llc Combinatorial lubricating oil composition libraries
JP2007514800A (ja) * 2003-10-31 2007-06-07 シェブロン・オロナイト・カンパニー・エルエルシー コンビナトリアル潤滑油組成物ライブラリ
US9068953B2 (en) 2007-09-17 2015-06-30 Agena Bioscience, Inc. Integrated robotic sample transfer device
WO2014052174A1 (fr) * 2012-09-25 2014-04-03 Merck Sharp & Dohme Corp. Diversification de composé à l'aide d'une fonctionnalisation au stade tardif

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