WO2001029537A2 - Method and apparatus for monitoring solid phase chemical reactions - Google Patents
Method and apparatus for monitoring solid phase chemical reactions Download PDFInfo
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- WO2001029537A2 WO2001029537A2 PCT/US2000/028218 US0028218W WO0129537A2 WO 2001029537 A2 WO2001029537 A2 WO 2001029537A2 US 0028218 W US0028218 W US 0028218W WO 0129537 A2 WO0129537 A2 WO 0129537A2
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- reaction
- total reflection
- attenuated total
- solid support
- reflection element
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/251—Colorimeters; Construction thereof
- G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
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- 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
- B01J2219/00279—Features relating to reactor vessels
- B01J2219/00306—Reactor vessels in a multiple arrangement
- B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
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- 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
- B01J2219/00279—Features relating to reactor vessels
- B01J2219/00306—Reactor vessels in a multiple arrangement
- B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
- B01J2219/00315—Microtiter plates
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- 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
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- 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
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- 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
- B01J2219/00686—Automatic
- B01J2219/00691—Automatic using robots
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- 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
- B01J2219/00693—Means for quality control
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- 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
- B01J2219/00695—Synthesis control routines, e.g. using computer programs
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- 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
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- 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
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- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
- C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries
Definitions
- the present invention concerns methods and instruments for monitoring chemical reactions, and particularly concerns methods and instruments for monitoring solid phase chemical reactions.
- the present invention provides a method for monitoring a solid phase chemical reaction.
- the method comprises the steps of: (a) providing a reaction mixture comprising a solid support and a liquid reaction medium, (b) contacting an attenuated total reflection element to said reaction mixture; and then (c) monitoring the chemical reaction on said solid support through said attenuated total reflection element.
- the monitoring step is carried out by attenuated total reflection spectroscopy.
- the chemical reaction on the solid support may be directly monitored, rather than indirectly monitoring that chemical reaction by monitoring reaction constituents in the liquid reaction medium.
- the contacting step is carried out under conditions that cause the progression of the desired reaction, with the appropriate reagents included in the reaction medium.
- the monitoring step is preferably carried out by measuring light absorbance by the reaction mixture, with changing light absorbance being positively associated with the progression of the reaction (e.g., increasing light absorbance being positively associated with the progression of a synthesis reaction, or decreasing light absorbance being positively associated with the progression of a degradation reaction).
- the foregoing method can be used to directly monitor the progression of chemical reactions as they proceed on solid phase supports.
- the progression of reaction steps on the bead can be monitored without having to remove a sample or cleave the molecule offofthe bead. While applicable to any type of small or large scale solid phase synthesis, this method is particularly useful for application to solid phase synthesis carried out to generate a solid phase combinatorial library, where the time required for each step to generate the library can be advantageously reduced.
- the beads e.g., infra-red, near infra-red, visible, or ultra-violet light absorbance
- a further aspect of the present invention is a method of making a combinatorial library preferably comprising at least 100 different compounds, with each of said compounds immobilized on a discrete solid support, by at least two sequential reaction cycles, and with each of said reaction cycles comprising a solid phase chemical reaction, wherein all of the sequential reaction cycles are completed in a total time that averages from not more than 8 or 10 hours for each of the sequential reaction cycles.
- Figure la is a schematic illustration of an apparatus that may be used to carry out the present invention.
- Figure lb is a detailed view of a segment of the apparatus of Figure la, showing the adhesion of the solid phase synthesis support beads to the attenuated total reflectance element.
- Figure lc is a detailed view of the a segment of the apparatus of Figure lb, showing the optical relationship of the solid support beads to the attenuated total reflectance element.
- Figure 2 illustrates a solid state Friedel-Crafts alkylation reaction that may be used in carrying out the present invention.
- Figure 4 illustrates a solid state Suzuki reaction that may be used in carrying out the present invention.
- Figure 5 illustrates a Mitsunobu reaction.
- Figure 6 illustrates a solid state Mitsunobu reaction that may be used in carrying out the present invention.
- Figure 7 illustrates the results obtained with a Friedel-Crafts reaction of the present invention.
- the upper box of this figure is a waterfall plot of absorbance versus time; the lower box of this figure is a contour plot over time.
- Figure 8 illustrates the wash steps carried out with the reaction of Figure 7, with a total of 15 washing steps being employed.
- the upper box of this figure is a waterfall plot of absorbance versus wash number. As can be seen there is a large initial absorbance with the trend towards zero absorbance as the washes progress.
- the bottom portion of the figure is a contour plot with the content of the wash indicated at the top of the plot and the wash progression proceeding from right to left. It can be seen from this plot that the first 5 washes remove a substantial amount of the excess reagent.
- Figure 9 illustrates the results obtained with a Suzuki coupling reaction with the present invention.
- Figure 10 illustrates the results for the first step of a Mitsunobu reaction in carrying out the present invention.
- Figure 11 illustrates the monitoring of chemistry on beads, or the solid phase, and not the reaction solution, in a Suzuki coupling reaction in the present invention.
- Figure 12 schematically illustrates a combinatorial library synthesis apparatus employing an attenuated total reflection monitor of the present invention.
- Figure 13a illustrates a probe and support structure arrangement for monitoring reactions in a plurality of reaction wells.
- Figure 13b illustrates a probe and support structure arrangement for monitoring reactions in a plurality of reaction wells.
- Figure 13c illustrates a probe and support structure arrangement for monitoring reactions in a plurality of reaction wells.
- the present invention is useful for all types of solid phase reactions, including batch reactions, single reactions, or reactions incorporated into a combinatorial synthesis process for making a combinatorial library.
- Combinatorial libraries are sets or collections of different compounds, which compounds are immobilized on a solid support, with different compounds immobilized on a different discrete solid support as described below.
- the compounds may be oligomers or nonoligomers, as also discussed below.
- the apparatus comprises a fiber optic coupled attenuated total reflection (ATR) probe 20 connected to a ultraviolet/visible light spectrograph 25.
- the ATR probe (obtained from Equitech International, Aiken, South Carolina, USA) was employed a three-bounce design attenuated total reflection element 22 and incorporated 400 ⁇ m fiber optic fibers as the cable interconnecting the spectrograph to the probe.
- Two different spectrographs were used for this work depending on the number of channels monitored: A single channel spectrograph using a deuterium light source (Zeiss MCS 501, Custom Sensors and Technology, St.
- the total internal reflection can be carried out with ultraviolet light, near-ultraviolet light, visible light or infrared light.
- the attenuated total reflection element may be formed from any suitable material that is substantially optically transparent at the wavelength employed, but is generally formed from an inert insoluble inorganic crystal material such as sapphire, glass, quartz, germanium, zinc selenide, diamond (including diamond like carbon), and combinations thereof.
- the attenuated total reflection element may be formed from composites or layers in accordance with known optical fabrication techniques, and may uncoated or coated with a material that will facilitate adhesion of the solid supports to the attenuated total reflection element
- the probe 20 is inserted into a solid phase reaction medium 30 comprising a liquid phase 31 and discrete solid supports, preferably polystyrene beads, as the solid support 32.
- the solid particles 32 adhere or bind to the ATR element 22, with light entering and leaving the element along paths 35, 36.
- light 37 from the ATR element 22 is believed to enter and penetrate the solid supports 32 due to the adhesion or binding of the solid supports 32 to the ATR element 22.
- the solid support is a separate discrete solid support such as a particle or bead
- a major portion, or substantially all of, the exposed surface of the attenuated total reflection element is coated with a continuous layer of said separate discrete solid support that has a thickness of at least one of said separate discrete solid supports, as illustrated in Figure lb.
- Solid supports used to carry out the present invention are typically discrete solid supports. Discrete solid supports may be separate from one another as described above, or may be discrete regions on a surface portion of a unitary substrate. Such "chip-type” or “pin-type” solid supports are known. See, e.g., U.S. Patent No. 5,288,514 to Ellman (pin-based support); U.S. Patent No. 5,510,270 to Fodor et al. (chip-based support) (the disclosures of all United States patent references cited herein are to be incorporated by reference herein in their entirety). Separate discrete supports such as particles or beads (these terms being used interchangeably herein), disks, fibers, needles or the like, are currently preferred.
- the discrete solid supports are formed from a polymer such as polystyrene.
- the solid substrates are beads, which may be completely solid throughout, porous, deformable or hard.
- the beads will generally be at least 10, 20 or 50 to 250, 500, or 2000 ⁇ m in diameter, and are most typically 50 to 250 ⁇ m in diameter.
- any convenient composition can be used for the particles or beads, including cellulose, pore-glass, silica gel, polystyrene beads such as polystyrene beads cross-linked with divinylbenzene, grafted copolymer beads such as polyethyleneglycol/polystyrene, polyacrylamide beads, latex beads, dimethylacrylamide beads, composites such as glass particles coated with a hydrophobic polymer such as cross-linked polystyrene or a fluorinated ethylene polymer to which is grafted linear polystyrene, and the like. Where separate discrete solid supports such as particles or beads are employed, they generally comprise from about 1 to 99 percent by weight of the total reaction mixture.
- the reaction medium apart from the solid support is, in general, a liquid.
- the reaction medium will comprise from about 1 to 99 percent by weight of the total reaction medium (where the solid support is separate from the container in which the reaction mixture is held, as in the case of particles or beads).
- Any suitable liquid may be employed, including aqueous liquids (water), nonaqueous liquids (e.g., organic solvents), and combinations thereof or mixtures thereof.
- the liquid may be a single phase or multi-phase solution.
- Organic solvents used to carry out the present invention may be polar or nonpolar, protic or aprotic, etc. The particular solvent or mixture of solvents depends upon the particular reaction being carried out.
- Suitable solvents for use in the reaction medium include, but are not limited to, methylene chloride, dimethyl formamide, 1 -methyl-2-pyrrolidinone , methanol, ethanol, water, toluene, tetrahydrofuran, dioxane, ethyl acetate, chloroform, acetone, acetic anhydride, pentane, hexane, benzene, carbon tetrachloride, diethyl ether, acetonitrile, etc.
- the reaction medium will include the reagents necessary to carry out the reaction desired, such as one or more reactants, a catalyst and/or initiator if necessary for the particular reaction, etc. All can be routinely determined by skilled artisans based on the particular reaction desired.
- the reaction mixture is held or placed in conditions that cause or permit the reaction to progress.
- the particular reaction conditions are not critical, depend upon the particular reaction desired, and can be routinely determined by skilled artisans.
- the reaction may be carried out under atmospheric pressure, elevated pressure, or reduced pressure; the reaction may be carried out at room temperature, elevated temperature, or reduced temperature; the reaction may be carried out at a neutral, acidic, or basic pH; the reaction may be carried out with or without an inert blanketing gas such as nitrogen or argon; etc.
- Any suitable reaction can be employed to carry out the solid-phase synthesis that is monitored or observed by the present invention. See, e.g., U.S. Patent No. 5,565,324.
- suitable reactions include, but are by no means limited to, Mitsunobu, Freidel-Crafts, Suzuki, Merrifield (for the synthesis of polypeptides)(.see, e.g., Merrifield, J. Am. Chem. Soc.
- reaction or reactions employed in carrying out the present invention may thus be selected to produce a variety of products, the progression of the synthesis of - o - which products can be monitored as described herein.
- Such products are, in general, non-oligomers, oligomers, or combinations thereof.
- Non-oligomer reaction products include a wide variety of organic molecules, such as heterocyclics, aromatics, alicyclics, aliphatics and combinations thereof, comprising steroids, antibiotics, enzyme inhibitors, ligands, hormones, drugs, alkaloids, opioids, terpenes, porphyrins, toxins, catalysts, as well as combinations thereof.
- Oligomer reaction products include oligopeptides, oligonucleotides, oligosaccharides, polylipids, polyesters, polyamides, polyurethanes, polyureas, polyethers, and poly (phosphorus derivatives), e.g.
- the present invention provides a direct monitoring technique rather than an indirect monitoring technique. By directly monitoring the progression of the reaction with the methods described herein, the reaction is advantageously monitored in real time as the reaction occurs.
- reaction need not be inferred from other measurements, and it is not necessary to have an intervening sampling, handling, or other processing step that intervenes between the reaction event or events and analysis. Further, the reaction need not be "opened” or otherwise disrupted as atmosphere, pressure, temperature, concentration, volume and/or stirring is changed by the sampling or processing step. As a result, individual solid state reactions can be efficiently and effectively monitored so that the reaction materials and equipment can be optimized for a particular combinatorial chemistry and synthesis program, allowing the time and effort required to carry out the combinatorial synthesis to be reduced.
- the present invention is primarily concerned with monitoring the progression of synthesis reactions, and while the object of a series of reactions may be to synthesize a compound, that a given reaction may involve the addition, removal (e.g., degradation), or substitution of a chemical unit from the core compound in solid phase, and that the progression of all such reactions may be monitored by the present invention.
- the method of the present invention can be employed to monitor single batch reactions, as described in connection with Figure 1 above, or can be used to monitor sequential and/or concurrent reactions, such as employed in the synthesis of a combinatorial chemical library, where the reaction product may be oligomers or nonoligomers as described above.
- the apparatus includes a chemical synthesis robot 101 configured to receive a plurality of reaction wells, as discussed in greater detail below.
- a reagent repository 102 is operatively associated with the chemical synthesis robot in accordance with standard techniques.
- An attenuated total reflection element as described above is operatively associated with the chemical synthesis robot for insertion into at least one, and preferably a multiplicity or even all of the reaction wells.
- An attenuated total reflection monitor 103 is connected to each attenuated total reflection element, which are configured to monitor a solid phase synthesis reaction in the corresponding reaction well by attenuated total reflection spectroscopy.
- a synthesis controller 104 is operatively associated with both the reagent repository and the chemical synthesis robot. The synthesis controller is configured through the implementation of hardware and/or software instructions to control the construction of a solid phase combinatorial library in the plurality of reaction wells. Synthesis robots, reagent repositories and controllers therefore are known in the art and can be implemented in accordance with standard techniques. See, e.g., U.S. Patent No.
- the synthesis controller is operatively associated with the attenuated total reflection monitor through an appropriate interface, with the synthesis controller configured through hardware and/or software instructions to end a particular synthetic step or reaction upon detection of completion of a chemical reaction, so that the overall combinatorial synthesis scheme can proceed to the next cycle of reaction steps.
- the reaction wells may be incorporated into the robot, but are typically provided in a separate structure that is insertable into or carried by the robot, as is known in the art. Microtiter plates, individual containers, columns, gels, Teraski plates, flasks, Merrifield synthesis vessels, etc., can be employed.
- the attenuated total reflection element can be associated with the reaction wells in a combinatorial synthesis apparatus as described in Figure 12 in any of a variety of ways, some of which are illustrated in Figures 13a-c.
- the well support structure 120a contains a plurality of reaction wells 121a.
- the bottom of each well is plugged and sealed by a probe 122a that terminates with the attenuated total reflection element 123a.
- the probes are thus connected to the robot stage with the support structure removably connected thereto, and with each well sealed by the reflection element.
- the contents of each well can be pipetted therefrom for subsequent reactions.
- Figure 13b An alternate approach is illustrated in Figure 13b, which again comprises a support structure 120b containing a plurality of reaction wells 121b.
- the total reflection elements 123b are rigidly and sealably connected into the support structure 120b, while the probe elements 122b are removably contacted to the reflection elements.
- the entire support structure can be simply secured through an appropriate stage to the probe elements, which again are connected to the synthesis robot.
- Figure 13c again comprises a support structure 120c having a plurality of reaction wells 121c, with a plurality of probes 122c, each carrying an attenuated total reflection element 123c, inserted into each reaction well.
- This structure can employ a conventional microtiter plate.
- the probes are carried by the synthesis robot, which can be structured so that the probes are placed in the reaction well before, during, or after the addition of supports, reaction medium or liquid, and other reagents into each well.
- the foregoing apparatus provides a method of making a combinatorial library by solid phase chemical synthesis carried out by the following steps: (a) combining (typically in a reaction well) a solid support, a liquid reaction medium and reaction reagents to produce a reaction mixture;
- step (e) repeating steps (a) through (d) with the separated solid supports produced in step (d) above.
- the repeating step is carried out cyclically until the desired number of compounds is produced as the combinatorial library.
- the library so produced may be comprised of oligomer or nonoligomer compounds, as described above, by any type of solid phase reaction, including but not limited to those described above.
- completion of the reaction is achieved when the reaction is sufficiently complete to achieve reasonable uniformity of the product population for a given library constituent, or group of constituents, to achieve the desired result of producing a useful combinatorial library.
- completion of the reaction occurs when the reaction reaches equilibrium.
- steps (a) through (d) above are concurrently carried out in a plurality of different reaction wells, and then a new set initiated in step (e), again typically in a plurality of different reaction wells.
- steps (a) through (d) may be cyclically repeated, individually or as sets of concurrent reactions, as many times as necessary to produce the desired combinatorial library, typically 2, 3 or 4 repetitions up to 10, 20, 30 or 40 repetitions or more.
- each reaction cycle may be concurrently carried out in 10, 20 or 30 up to 400, 600 or 800 or more wells or reaction vessels.
- Current formats include 96 well and 384 well microtiter plates, but it is not necessary that every well in the plate be used.
- diversity in the library can be achieved by any means of solid phase or solid state synthesis, such as by carrying out different series of syntheses in different wells, by pooling the solid supports from multiple different reaction wells (including some or all) after a synthetic cycle, splitting that pool back into different reaction wells for a further set of reactions, and then repeating that cycle, in accordance with known techniques.
- any means of solid phase or solid state synthesis such as by carrying out different series of syntheses in different wells, by pooling the solid supports from multiple different reaction wells (including some or all) after a synthetic cycle, splitting that pool back into different reaction wells for a further set of reactions, and then repeating that cycle, in accordance with known techniques.
- Reactions, reaction conditions and reaction products include but are not limited to those described above.
- combinatorial libraries containing at least 100, 500, 1,000 or 5,000 different compounds up to 10,00, 20,000, 50,000, 100,000, up to 10 6 different compounds or more, can be produced more rapidly than by prior techniques.
- the library may be relatively large or relatively small, but the present invention is useful in the preparation of both relatively small libraries and relatively large libraries on a more expeditious basis.
- the process of synthesizing a combinatorial library may be varied from that described above.
- the attenuated total reflection spectroscopy step may be carried out in situ during the chemical reaction cycle of interest, and completion directly detected.
- the attenuated total reflection spectroscopy step may be carried out a priori on a model reaction system prior to the chemical reaction cycles of interest to, the reaction optimized and a necessary reaction time determined.
- the reaction cycle that incorporates a reaction step based upon the reaction model (and for which a reaction time has been determined a priori) may be terminated upon completion of the previously determined reaction time.
- a plurality of, a majority of, or all of the reaction cycles may be carried out to completion, with completion being determined by either in situ measurement or a priori measurement as described above, or combinations thereof. It will also be appreciated that not every reaction in a particular combinatorial synthesis need by optimized by the techniques described herein, so long as the overall library synthesis time is advantageously reduced.
- a time to completion is determined a priori from a model reaction
- that model reaction can be the same as the reaction carried out in the synthesis of the combinatorial library, or may differ in one or more parameters, with the reaction time for the combinatorial synthesis reaction being interpolated from the model reaction.
- a time to completion is interpolated from a different reaction, it is preferably interpolated from a plurality of different reactions in accordance with known techniques.
- the time to synthesize the library can be substantially reduced.
- all of the reaction cycles may be completed in a time that averages not greater than 6, 8 or 10 hours per reaction cycle (typically, an average time of at least 1 or 2 hours per reaction cycle is employed).
- the average reaction time per cycle is 4 hours. Note that in determining the average time any resting time or periods of inactivity between cycles is not counted, but the time for all aspects of each reaction, including mixing and washing steps, is included.
- the compounds on the solid supports or the solid supports themselves may be tagged for later decoding by any suitable technique in accordance with known procedures, including but not limited to oligonucleotide tags and oligopeptide tags, see, e.g., S. Brenner and R. Lerner, Proc. Natl. Acad. Sci. USA 89, 5381 (1992); J. Kerr et al., J. Am. Chem. Soc. 115, 2529 (1993), nonsequential tags, such as described in U.S. Patent No. 5,565,324 to Still, or mass-based encoding such as described in PCT Application WO97/37953 to Geysen et al. Tagging reactions may be carried out concurrently with the reactions employed to synthesis the members of the combinatorial library in the same reaction medium.
- a Friedel-Crafts reaction is an electrophilic substitution
- the general reaction steps are as follows (r. Morrison and R. Boyd, Organic Chemistry (Allyn and Bacon, Inc., 1973):
- the electrophile is typically a carbonium ion and is formed in an acid-base equilibrium, as illustrated in Scheme 1 below:
- Suzuki couplings involve the palladium-catalyzed couplings of unsaturated halides with boronic acids or esters. Suzuki coupling reactions are known in the art. See, e.g., G. Smith et al., J. Org. Chem. 59, 8151 (1994). The general Suzuki coupling is shown in Figure 3. An example of this type of reaction applied to solid phase synthesis is shown in Figure 4. In brief, in an illustrative reaction, precursor resin, 0.4 grams, is suspended in 6 mL of methylene chloride and stirred.
- Mitsunobu reactions refer to the reaction of hydroxy compounds with acids (pK, ⁇ 11) in the presence of triphenylphosphine and diethyl azodicarboxylate. See, e.g., M. Narasi et al., J. Org. Chem. 52, 4235 (1987). This scheme is used for the functionalization of alcohols and related compounds.
- the general Mitsunobu reaction is shown in Figure 5.
- a specific solid phase Mitsunobu reaction is shown in Figure 6.
- Sasrin resin 2.16 grams, is suspended in 50 mL of tetrahydorfuran and gently agitated for 30 minutes under a blanket of nitrogen.
- ⁇ - hydroxyphthalimide 1.60 grams, and triphenylphosphine, 2.52 grams, is added and the mixture is agitated until these reagents are dissolved.
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
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EP00970831A EP1221037A2 (en) | 1999-10-15 | 2000-10-12 | Method and apparatus for monitoring solid phase chemical reactions |
JP2001532079A JP2003512615A (en) | 1999-10-15 | 2000-10-12 | Method and apparatus for monitoring solid phase chemical reactions |
AU80154/00A AU8015400A (en) | 1999-10-15 | 2000-10-12 | Method and apparatus for monitoring solid phase chemical reactions |
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US15967399P | 1999-10-15 | 1999-10-15 | |
US60/159,673 | 1999-10-15 |
Publications (2)
Publication Number | Publication Date |
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WO2001029537A2 true WO2001029537A2 (en) | 2001-04-26 |
WO2001029537A3 WO2001029537A3 (en) | 2001-11-29 |
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PCT/US2000/028218 WO2001029537A2 (en) | 1999-10-15 | 2000-10-12 | Method and apparatus for monitoring solid phase chemical reactions |
Country Status (4)
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EP (1) | EP1221037A2 (en) |
JP (1) | JP2003512615A (en) |
AU (1) | AU8015400A (en) |
WO (1) | WO2001029537A2 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007041999A1 (en) * | 2005-10-10 | 2007-04-19 | Johann Wolfgang Goethe-Universität Frankfurt am Main | Device for qualitatively and/or quantitatively determining ir active constituents in liquids and a method for qualitatively and/or quantitatively determining ir active constituents in liquids |
US8390816B2 (en) | 2008-03-04 | 2013-03-05 | Kurashiki Boseki Kabushiki Kaisha | Method for attenuated total reflection far ultraviolet spectroscopy and an apparatus for measuring concentrations therewith |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7283245B2 (en) * | 2004-01-20 | 2007-10-16 | General Electric Company | Handheld device with a disposable element for chemical analysis of multiple analytes |
JP4765059B2 (en) * | 2005-02-10 | 2011-09-07 | 国立大学法人北海道大学 | Cognitive task response measurement system and cognitive task response measurement method |
JP5982663B2 (en) * | 2011-12-13 | 2016-08-31 | 株式会社Ube科学分析センター | Infrared spectrometer for measuring electrolyte of lithium ion secondary battery and measuring method thereof |
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US4582809A (en) * | 1982-06-14 | 1986-04-15 | Myron J. Block | Apparatus including optical fiber for fluorescence immunoassay |
US5313264A (en) * | 1988-11-10 | 1994-05-17 | Pharmacia Biosensor Ab | Optical biosensor system |
WO1994019694A1 (en) * | 1993-02-19 | 1994-09-01 | Arris Pharmaceutical Corporation | Thin film hpmp matrix systems and methods for constructing and displaying ligands |
US5565324A (en) * | 1992-10-01 | 1996-10-15 | The Trustees Of Columbia University In The City Of New York | Complex combinatorial chemical libraries encoded with tags |
US5780603A (en) * | 1996-11-15 | 1998-07-14 | Synsorb Biotech, Inc. | Combinatorial synthesis of carbohydrate libraries |
US5817751A (en) * | 1994-06-23 | 1998-10-06 | Affymax Technologies N.V. | Method for synthesis of diketopiperazine and diketomorpholine derivatives |
WO1999004228A2 (en) * | 1997-07-16 | 1999-01-28 | Ljl Biosystems, Inc. | Light detection device |
US5942387A (en) * | 1996-08-26 | 1999-08-24 | Eli Lilly And Company | Combinatorial process for preparing substituted thiophene libraries |
US5965719A (en) * | 1996-11-15 | 1999-10-12 | Sunsorb Biotech, Inc. | Combinatorial synthesis of carbohydrate libraries |
-
2000
- 2000-10-12 EP EP00970831A patent/EP1221037A2/en not_active Withdrawn
- 2000-10-12 WO PCT/US2000/028218 patent/WO2001029537A2/en active Search and Examination
- 2000-10-12 AU AU80154/00A patent/AU8015400A/en not_active Abandoned
- 2000-10-12 JP JP2001532079A patent/JP2003512615A/en active Pending
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US4582809A (en) * | 1982-06-14 | 1986-04-15 | Myron J. Block | Apparatus including optical fiber for fluorescence immunoassay |
US5313264A (en) * | 1988-11-10 | 1994-05-17 | Pharmacia Biosensor Ab | Optical biosensor system |
US5565324A (en) * | 1992-10-01 | 1996-10-15 | The Trustees Of Columbia University In The City Of New York | Complex combinatorial chemical libraries encoded with tags |
WO1994019694A1 (en) * | 1993-02-19 | 1994-09-01 | Arris Pharmaceutical Corporation | Thin film hpmp matrix systems and methods for constructing and displaying ligands |
US5817751A (en) * | 1994-06-23 | 1998-10-06 | Affymax Technologies N.V. | Method for synthesis of diketopiperazine and diketomorpholine derivatives |
US5942387A (en) * | 1996-08-26 | 1999-08-24 | Eli Lilly And Company | Combinatorial process for preparing substituted thiophene libraries |
US5780603A (en) * | 1996-11-15 | 1998-07-14 | Synsorb Biotech, Inc. | Combinatorial synthesis of carbohydrate libraries |
US5965719A (en) * | 1996-11-15 | 1999-10-12 | Sunsorb Biotech, Inc. | Combinatorial synthesis of carbohydrate libraries |
WO1999004228A2 (en) * | 1997-07-16 | 1999-01-28 | Ljl Biosystems, Inc. | Light detection device |
Non-Patent Citations (1)
Title |
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BIRNBAUM S ET AL: "PEPTIDE SCREENING" CURRENT OPINION IN BIOTECHNOLOGY,GB,LONDON, vol. 3, 1992, pages 49-54, XP000651934 ISSN: 0958-1669 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007041999A1 (en) * | 2005-10-10 | 2007-04-19 | Johann Wolfgang Goethe-Universität Frankfurt am Main | Device for qualitatively and/or quantitatively determining ir active constituents in liquids and a method for qualitatively and/or quantitatively determining ir active constituents in liquids |
US8390816B2 (en) | 2008-03-04 | 2013-03-05 | Kurashiki Boseki Kabushiki Kaisha | Method for attenuated total reflection far ultraviolet spectroscopy and an apparatus for measuring concentrations therewith |
Also Published As
Publication number | Publication date |
---|---|
WO2001029537A3 (en) | 2001-11-29 |
EP1221037A2 (en) | 2002-07-10 |
AU8015400A (en) | 2001-04-30 |
JP2003512615A (en) | 2003-04-02 |
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