JP2005503370A - Method and apparatus for directed selective combinatorial synthesis - Google Patents

Abstract

Selection apparatus and methods for directed selective combinatorial chemical synthesis are provided. The present invention relates to a directed and flexible selection apparatus and method for combinatorial chemical synthesis using directed splitting and recombination processes. This process utilizes a single algorithm that is universal for the orientation and selection of libraries of any size. The invention further provides for releasing one or more supports organized in a one-dimensional array in one or more reaction vessels into an isolation and transfer chamber, wherein the one or more supports are said reaction vessels. Each in a patterned distribution to one or more subsequent reaction vessels, wherein the patterned distribution of supports in each subsequent reaction vessel is the same, and in the reaction vessel Each support location provides a method of selecting a solid support for combinatorial chemical synthesis that encodes its previous synthesis history. In another aspect, the invention provides a directed selection device in which all members of a set at a given time are reacted with the same chemical building block throughout synthesis.

Description

【Technical field】
[0001]
(Background technology)
Protocols have recently been developed for combinatorial synthesis in which a large number of compounds are synthesized individually. These methods proceed by a sequence of steps where each step is added to a plurality of building blocks, ie a specific selected one of small organic molecules, to the growing intermediate compound. The number of possible final compounds thereby is a product of mathematical terms, and each synthesis step term represents the number of possible building blocks that can be added in that step.
[0002]
In combinatorial synthesis, the addition step reaction typically proceeds by combining a partially synthesized intermediate compound with a building block that has an attached activated residue. (Hereafter, it is assumed that the building block has the necessary activated residues attached.) Activation reagents and other reagents and solvents are also added to the addition step reaction. Building blocks are often added in molar excess over the partially synthesized compounds present so that the thermodynamically favorable building block addition reaction is substantially complete. After the addition of one building block, the intermediate compound is separated from the spent reaction solution and prepared for further building blocks. The intermediate compound is often attached to the solid support by a linking group that can be cleaved, for example, to simplify its separation from the spent reaction. In such a solid phase protocol, the final step of cleaving the linking group releases the final compound.
[0003]
Building blocks (activated as needed), activation and other reagents, and reaction conditions have recently been completed for many classes of final compounds. Examples of such reactions and protocols are the following for the addition of natural and artificial amino acids to produce peptides.
Lam et al. , 1991, Nature 354: 82-84 “A new type of synthetic peptide library for identifying ligand binding activity”.
Lam et al. , US Pat. No. 5,510,240 “Peptide Library Screening Method”
Lam et al. , 1994, Selectide Technology 6: 372-380 “Bead Binding Screening Method: Comparison with Enzymatic Methods”
[0004]
See, for example, the following for protrols for the synthesis of benzodiazepine compounds.
Bunin et al. 1992, J. Am. Amer. Chem. Soc. , 114: 10997-10998 “General and Convenient Method for Solid-Phase Synthesis of 1,4-Benzodiacepine Derivatives”
Ellman US Pat. No. 288,514 “Solid-phase combined synthesis of benzodiazepine compounds on solid phase”
See also, for example, for protocols for the addition of N-substituted glycines to produce peptoids.
Simon et al. , 1992, Proc. Natl. Acad. Sci. USA, 89: 9367-9371 “Modular Approach to Drug Discovery”
Zuckermann et. al. 1992, J. Am. Amer. Chem. Soc. 114: 10646-10647 “Efficient method for the preparation of peptoids [oligo (N-substituted glycines)] by submonomer solid phase synthesis”
Moos et al. , WO 94/06451 “Synthesis of N-substituted polyamide monomers useful as solvents, food additives, enzyme inhibitors, etc.”
[0005]
Recently, approaches for the synthesis of small molecule libraries have been summarized, for example, in the following literature.
Krchnak et al. , 1669, Molecular Diversity, 1: 193-216-216 “Synthetic Library Technique: Subjective Thinking”
Ellman, 1996, Account, Chem. Res. 29: 132-143 “Design, synthesis and evaluation of small molecule libraries”
Armstrong et al. , 1996, Account, Chem. . Res. , C29: 123-131 “Multiple Component Strategy for Combinatorial Library Synthesis”
Fruchtel et al. , 1996, Angew. Chem. Int. Ed. 35: 17-42 “Chemistry on Solid Supports”
Thompson et al. , 1996, Chem. Rev. 96: 555-600 “Synthesis and application of small molecule libraries”
Rinnova et al. , 1996 Collect. Czech. Chem. Commun. 61: 171-231 “Molecular Diversity and Structure Library”
Hermkens et. al. Tetrahedron. 52: 4527-4554 “Solid-phase organic reactions: a review of recent literature”
[0006]
The predictable synthesis of a large number of individual compounds that are later collected in a library of compounds is of interest and useful in several fields, especially in drug discovery, agriculture, health, veterinary medicine, nutritional chemistry and material chemistry libraries . Identifying active compounds that provide structural information for use in the development of approved or commercially available therapeutic compounds and compound series validates the function of the molecule and makes the medicinal group (active molecular structural element) Of particular interest is the establishment of structure-activity relationship data to identify and identify active compounds, particularly in the field of pharmaceutical example compound selection. Pharmaceutical example compounds for certain drugs exhibit specific biological activities of pharmaceutical interest and are in addition to specific biological activities, pharmacological and suitable for administration to humans or animals. A compound that can serve as a starting point for the selection and synthesis of drug compounds having toxicological properties. It is clear that the synthesis of a large number of compounds and the screening of their biological activity in a controlled biological system can help in the selection of example compounds. Instead of turning to plants or other natural resources, medicinal chemists can use combinatorial protocols to create laboratory compounds to screen for the desired activity. See, for example, Borman, Chemical & Engineering News, Feb. 12, 1996, 29-54 "Combined chemists focus on small molecules, molecular recognition and automation."
[0007]
In order to benefit from these developed combination protocols, automated synthesizers are beneficial. Manual handling of hundreds, thousands, or perhaps tens of thousands of separate compounds is expensive, time consuming, and error prone. Therefore, synthetic robots have recently been developed for the automation of one or more steps of combinatorial protocols. An example of such a recently developed robot is Cargil et al. , 1996, Automated Combinatory Chemistry on Solid-Phase, Laboratory Robotics and Automation, 8: 139-148; Sugarman et al. , U. S. Pat. 5, 503, 805; Zuckermann et al. , U. S. Pat. 5,252,296, Methods and apparatus for biopolymer synthesis; Zuckermann et al. , WO 93 / 12,427, Automated apparel for use in peptide synthesis; Krchnak et al. , MARS-Multiple Automatic Robot Synthesizer-Continuous Flow of Peptide, Peptide Res. 9: 45-49.
[0008]
The most efficient and least demanding combinatorial chemistry method for the synthesis of multi-compound arrays was introduced by Arpad Furka (Furka et al. 1988b; Furka et al. 1988a; Furka et al. 1991). And later used independently by Kit Lam (Lam et al. 1991) and Richard Houghten (Houghten et al. 1991). This method is subject to limitations. For example, the amount of compound synthesized is limited by the loading of the solid support particles, and the chemical history of each particle must be recorded or determined by analytical methods (Lam et al., 1997).
[0009]
Split combinatorial methods develop on resin beads, and they typically support nanomolar amounts of material at best. Two different routes were implemented to increase the amount of compound. Richard Houghten invented a T-bag (Houghten 1985) in which resin beads are enclosed in a permeable container. IRORI MicroKans is based on the same idea. Another choice is M.M. H. Applying a microscopic group support introduced by Geysen (Geysen et al., 1984; Geysen et al., 1986; Geysen et al., 1987) and Ronald Frank (Frank and Doring, 1988). In order to increase loading and improve reaction kinetics, polypropylene moldings were grafted with polystyrene to produce SynPhase with a large surface area and very convenient shape and immediately followed by SynPhase lanthanum. The reaction kinetics and product yield from lanthanum are comparable to resin beads, but the handling of lanthanum provides unique benefits especially for the synthesis of fairly large libraries containing mg amounts of each compound.
[0010]
Various methods were used to identify each compound after synthesis. After each combination step, the chemical history lost due to mixing of individual particles is addressed by chemical coating on the beads (Kerretal., 1993; Nikolaev et al., 1993; Ohlmyer et al., 1993), T-bags were manually labeled (Houghten, 1985), and radio frequency tags were attached to MicroKans or lanterns (Moran et al., 1995; Nicolav et al., 1995). Recently (Smith et al., 1999) and others (Furka et al., 2000; Furka 2000) have described one-dimensional position tracking of solid-phase particles called necklace coding.
[0011]
In general, solid phase synthesis is a well-known procedure, reaction vessels are typically conventional, and solvents and reaction temperatures can range from -100 to + 200 ° C, and the reaction is complete. Inexpensive commercial containers, microtiter plates, syringes, etc. that can withstand the reaction conditions used in synthesis protocols that can take minutes or even days. In a preferred embodiment, the reaction vessel is a threaded teflon tube surrounded on both sides by luer fittings. The reaction vessel train is sealed with various sealing means. However, the reaction vessel can be tailored to withstand any harsh reaction conditions (ie high or low temperature, high or low pressure, or a closed system).
[0012]
Solid phase combinatorial synthesis typically proceeds according to the following steps. In the first step, the reaction vessel is charged with one or more solid supports and a first of a plurality of building blocks or linkers is covalently linked to the solid phase. Subsequent building block addition steps are performed, all of which involve iterative execution of the following substeps in the sequence chosen to synthesize the desired compound. Initially, a sufficient amount of solution containing the building block selected for addition is added to the reaction vessel such that the building block is present in a molar excess relative to the intermediate compound. The reaction is initiated and facilitated by activating reagents and other reagents and solvents that are also added to the reaction vessel. The reaction vessel is then incubated for a period of time, typically minutes to 24 hours, sometimes days, at a controlled temperature sufficient to allow the building block addition reaction to proceed to substantial completion. Optionally, the reaction vessel can be intermittently stirred during this incubation. Finally, in the final building block addition substep, the reaction vessel containing the solid support with the intermediate compound attached is prepared for the addition of the next building block by removal and reconditioning of the spent reaction solution. The Optionally, during this addition step, multiple building blocks can be added to a single reaction vessel to perform two or more synthesis steps on a single solid support. After the desired number of building block addition steps, the final compound is attached to a solid support and is present in the reaction vessel. The final compounds can be attached directly to their synthetic support and used directly, or alternatively cleaved from the support. In the latter case, the linker attaching the compound to the support is cleaved and the library compound is extracted.
[0013]
The advent of combinatorial chemical technology has led to the need to simultaneously handle a number of different solid phases (supports) with bound intermediates. Module solid phase supports are particularly useful for combinatorial arrays of compounds because of their advantageous physicochemical properties and the possibility of applying a directed selection approach that distributes the support during the combinatorial synthesis step. It has become popular for synthesis. These methods can also be applied to select a container that holds a resin or solution phase where the container is treated as a single support. To identify each support during combinatorial synthesis on lanterns, others have described the necklace coding principle. The necklace can be a string, wire, rod or instrument inserted into an aligned hole in each subject set. In order to select a lantern prior to the subsequent combination step, Applicants treat lanterns packed in tubes as a result of the directed selection device described in this disclosure, particularly compared to lantern strings. Found to provide benefits.
DISCLOSURE OF THE INVENTION
[0014]
The present invention relates to a selection apparatus and method for directed selective combinatorial chemical synthesis, and more particularly directed and flexible selection apparatus and method for combinatorial chemical synthesis using directed splitting and recombination processes. About. This process utilizes a universal algorithm for directing and selecting libraries of any size.
[0015]
In one aspect, the present invention discharges one or more supports organized in a one-dimensional linear array into one or more reaction vessels into an isolation and transfer chamber, each of the reaction vessels. Transferring one or more supports from one to one or more subsequent reaction vessels in a patterned distribution, wherein the patterned distribution of the supports in each subsequent transfer is the same , And the location of each support in one reaction vessel provides a method of selecting a solid support for combinatorial chemical synthesis where the previous synthesis history is encoded.
[0016]
In another aspect, the invention relates to two or more reaction vessels for holding one or more target sets organized in a first ordered pattern, and subjects from each of the reaction vessels. An isolation and transfer chamber for directed redistribution of the target set to one or more subsequent reaction vessels to receive the set from the isolation and transfer chamber, and a step for each building block added A directed selection apparatus for performing a combined solid phase synthesis of compounds, including iterating.
[0017]
In another aspect, the invention performs a reaction in each of one or more first reaction vessels to hold one or more sets of objects organized in a one-dimensional linear array. And releasing the target set from the reaction vessel to the isolation and transfer chamber for directed redistribution of the target set to one or more subsequent reaction vessels for receiving from the isolation and transfer chamber A directed selection apparatus for performing a combined solid phase synthesis of compounds comprising repeating the steps for each building block.
[0018]
In another aspect, the present invention provides a directed selection device in which all members of a set of subjects are reacted with the same chemical building block throughout the synthesis at a given point in time.
[0019]
The directed combinatorial synthesis of the present invention is a method for split mixed chemical synthesis. A single compound per support or multiple compounds per support can be used to produce large quantities of each compound with great efficiency, simplicity and speed. The solid support can be released for reaction in a flow-through tube or for handling in a microplate. The selection device follows an algorithm that is universal for directed selection of any library selection regardless of size. A single support can be selected at a time, or multiple supports, ie, subject sets, can be used to increase the scale / yield of each compound synthesized. The problem of chemical history lost due to the mixing of individual supports after each combination step is addressed by preserving the support sequence.
[0020]
In addition to solid phase synthesis, through the remixing process, this algorithm can be used to select any target including a vessel for liquid phase synthesis, or even to control a valve for directing reagents to a specific reaction vessel. Can be used. The use of a single universal algorithm only requires the chemist to enter the library design (ie the number of steps and the number of building blocks at each step). A computer following this algorithm can then design the synthesis and synthesis batch size. All building blocks can be validated, allowing increased quality control.
[0021]
Combinatorial chemical synthesis protocols direct the stepwise sequential addition of building blocks to partially synthesized intermediate compounds to synthesize final compounds. These protocols are generally divided into liquid phase protocols and solid phase protocols. In a liquid phase protocol, the final compound is synthesized in solution. In solid phase synthesis, the final compound is synthesized attached to a solid support that allows the use of simple mechanical means to separate the intermediate or partially synthesized intermediate compound during the synthesis step. The A preferred solid support for the present invention comprises SynPhase lanthanum, which can optionally be functionalized to attach the intermediate compound covalently. However, any support such as made from various glasses, plastics or resins can be used.
[0022]
Directed selective combinatorial synthesis provides a low-cost alternative to radiolabels. This method enables individual solid phase particles to be tracked or directed to the efficiency of the fission-mixing approach, for example, the position of the support itself through each step of the synthesis and the final distribution to the vessel for cleavage and compound extraction / storage. Combines the convenience of recording chemical histories.
[0023]
In further embodiments, the invention consists of other combinations and subcombinations of the elements described above that function in combination with other devices or independently of any device. Directed combinatorial synthesis is based on the directed selection of supports or reaction vessels as they are moved, or in other embodiments they are not moved but rather on the support or reaction vessel where the addition of reagents is commanded. Or a combination of both. One skilled in the art can foresee many ways to control the release of the reagent to the position described by the algorithm (eg, control of a valve that commands the addition of the reagent to a particular reaction vessel). In a further embodiment, the reaction vessel array, eg, a three-dimensional array, eg, FIG. 8, is separated from the nearest reaction vessel by a valve. Recognizing that a similarly functioning array can also be constructed by reaction vessels not physically in one array but connected by tubes and valves to create the same functional connection between sets of reaction vessels. Should.
[0024]
A person skilled in the art can index the receiving container instead of indexing the releasing container and fixing the receiving container, and fix the discharging container or other mechanical, valve control or support transport means the same It can be appreciated that it can be used to obtain directed selection results and that the isolation and transfer chambers can be replaced by other support transport means such as valves or electromotive transport of the support in a microfluidic system.
[0025]
Similarly, instead of using a support, one of ordinary skill in the art would be able to use multiple solid phases such as an actual reaction vessel in which a solution phase reaction takes place, or a MicroKan or closed reaction vessel accessed through a stopper included in the reaction vessel, for example. It can be seen that a reaction vessel containing resin particles can be used.
[0026]
Throughout this disclosure, the following terminology has the following conventional meaning.
[0027]
A “chemical library” or “combination library” or “compound library” or “array” is the intentional creation of different compounds, usually prepared in parallel and screened for biological activity in a variety of different formats. Collection.
[0028]
“Building block” refers to any molecule that can be covalently attached to another molecule to produce a structurally different compound, generally a small organic molecule. Addition of chemical building blocks is often referred to as subunit addition or substitution addition.
[0029]
“Combined chemistry” or “combined synthesis” refers to a directed strategy for the parallel synthesis of various molecules leading to the generation of chemical libraries. This strategy consists of covalently connecting sequential structurally different building blocks to each other sequentially and repeatedly to obtain a large array of compounds.
[0030]
“Linker” refers to a molecule or group of molecules covalently attached at one end to a solid phase and at the other end to a first building block. Linkers have different molecular structures and therefore have different lengths, shapes, sizes, degrees of hydrophobicity and hydrophilicity, steric bulk, and chemical reactivity. The choice of linker in solid phase synthesis depends on the synthesis scheme and the biological screening format.
[0031]
“Solid support” refers to a material or group of materials having a rigid or semi-rigid surface, appropriate size, shape and porosity, and high chemical resistance. Examples of solid phases are glass, silica, cellulose, polystyrene crosslinked with divinylbenzene, polystyrene-polyethylene glycol copolymer, silica gel, alumina gel, plastic, polyamide, polyimide, polytetrafluoroethylene, phenol formaldehyde polymer, poly (chlorotrimethyl). Fluoroethylene), halogenated polyolefins, and other support materials commonly used in peptide, polymer and small molecule solid phase synthesis.
[0032]
“Resin” refers to a solid support material grafted with a linker for attachment of a first building block. Examples of preferred resins are IRORI MicroKans, Wang resin (polystyrene resin with 4-alkoxybenzyl alcohol linker) and Sasrin resin (polystyrene resin with 2-methoxy-4-alkoxybenzyl alcohol linker). Other preferred resins are NovaBiochem, La Jolla, Calif. Published by Combinatorial Chemistry & Solid Phase Organic Chemistry Handbook, Solid Phase Science, San Rafael, Calif. It is described in the published Solid Phase Science catalog or the Rapp Polymers catalog issued by Rapp Polymer GmbH, Tubingen, Germany.
[0033]
“Reaction vessel” refers to a vessel in which a reaction takes place. In some cases it can act as a holder for the “solid support”. The reaction vessel can resist the solvents and reaction conditions used in the synthesis protocol.
[0034]
A “subject set” also refers to one or more solid supports, resins, or reaction vessels in the case of liquid phase synthesis that share the same chemical history. The members of the target set move together through the combined chemical synthesis process. A target set may be just one support, container or resin can, or if multiple scales are desired to increase the yield of each compound produced, multiple supports, containers or resin cans can be used. Can be used. Each member of the subject set will have the same number and order of building blocks, resulting in the same final compound.
[0035]
These and other features and benefits of the present invention will be better understood by reference to the accompanying drawings and the following description.
BEST MODE FOR CARRYING OUT THE INVENTION
[0036]
For clarity of disclosure and not limitation, a detailed description of the invention will now be provided with reference to the figures illustrating preferred embodiments of elements of the invention. However, the present invention includes alternative embodiments of these elements that perform similar functions in a similar manner apparent to those skilled in the art from the provided disclosure. In addition, the present invention is disclosed with respect to its preferred application to solid phase combinatorial chemical synthesis. The present invention is not so limited and includes the application of the various disclosed elements to other chemical protocols that also have similar functional steps that will be apparent to those skilled in the art. For example, the components of the present invention can be applied to appropriate liquid phase combinatorial chemical synthesis, to other solid phase chemical protocols, or any combination thereof.
[Selection algorithm]
[0037]
The present invention applies (i) the same algorithm before any combination step of synthesis, (ii) regardless of the number of building blocks in each step, and (iii) synthesizes any size library In order to do so, a general selection protocol can be employed that allows for the dispensing of standard volumes (or lengths) and multiple uses of receiving tubes. For example, lanterns can be organized into a one-dimensional or linear array in each distribution tube (FIG. 2). The first receiving tube has n receiving tubes _x = N _tot / BB _x The lanterns from the distribution tube are filled by transferring one lantern at a time from each of the distribution tubes until it contains lanthanum (FIG. 3). Where n _x Is the number of lanterns in the tube for the xth combination step, n _tot Is the total number of compounds (this number is equal to the number of lanthanums unless multiple supports per compound are used to increase the amount of compound produced) and BB _x Is the number of building blocks in the xth step. This algorithm is described schematically with an example of a library synthesized in a three-stage combinatorial step that uses 2, 3 and 4 building blocks in the first, second and third steps, respectively (FIG. 2). Rather than transfer directly between reaction vessels, the target set column is transferred onto a necklace (ie, string, wire, rod, or instrument inserted through aligned holes in each target set) and then the reaction vessel's The next set can be transferred, or the necklace can be used to transfer the subject set from the reaction vessel to the distribution device.
[0038]
This algorithm can be applied to directed selection by any process including the directed selection device described in this disclosure. In the example algorithm of FIG. 2, two tubes (or threaded necklaces) are depicted, each representing a representative “A” (light) or “B” (dark) building block, ie, reagent or building. Contains 12 lanthanum that is reacted with the block. Then all the lanterns are jumbled onto three new necklaces. One lantern was taken from each distribution tube and placed in the receiving tube (the last lantern from the first tube ends at the bottom of the first receiving tube and the last lantern in the second tube is the first End up as the second lantern from the bottom of the receiving tube, etc.). A total of 8 lanterns are transferred to the first receiving container. The same process is repeated twice more. The result was three new tubes containing the same array of lanterns.
[0039]
Each of these three tubes is reacted with the appropriate building block (eg, “C”, “D” or “E”) and then re-mixed using the exact same algorithm as before. The result is four new tubes each containing the same array of self-generated lanterns, which then each have four building blocks (eg, representative “F”, “G” ”,“ H ”and“ I ”). This process results in all 24 “letter” combinations, and the position in the last four tubes (or on the necklace if used) is undoubtedly a chemical history (each step in the process). The type of building block that reacted with lanthanum), or the putative structure synthesized on each lanthanum. FIG. 2a depicts the relationship of building blocks (eg characters) as the lantern moves through the synthesis step.
[0040]
In the example algorithm of FIG. 12, each of the representative “A”, “B” or “C” building blocks, ie, three tubes (or threaded) containing 9 lanthanum that is reacted with the reagent or building block. Necklace) is drawn. All lanterns are then remixed in 3 new tubes (or on 3 new necklaces). A total of 9 lanterns are transferred to the first receiving tube. The same process is repeated twice more. The result was three new tubes containing the same array of lanterns.
[0041]
These three tubes are again reacted with the appropriate building block (eg “D”, “E” or “F”) respectively and then the mess is repeated using the exact same algorithm as before. The result is three new tubes, each containing the same array (object set) of lanterns, which are then each possible three building blocks (eg representative “G”, “H” and “I”) ) And the last combination step. This process results in all 27 “letter” combinations, and the position of any lantern within the last three tubes (or on the necklace if used) is the chemical history (lanthanum at each step of the process). The type of building block reacted with) or the putative structure synthesized on each lanthanum is undoubtedly determined. The lanterns (target sets) in FIG. 11 are numbered to indicate the correlation between the positions of the lanterns after each jumble. FIG. 12 further depicts the relationship of building blocks (eg characters) as the lantern moves through the synthesis step.
[0042]
The tube-to-tube selection provides a handling benefit compared to selection on a string or necklace. For this reason, a selection tube can be used as a reaction vessel. In addition to being compatible with the directed selection device described in this disclosure, tubes have been recognized as highly efficient for many years (Lukas et al, 1981; Pryland and Sheppard 1986; Krchnak et al., 1987; Frank and Doring, 1988), and provides a convenient reaction vessel that has the benefit of using a continuous flow method for washing resin beads used in commercial synthesis.
[0043]
(apparatus)
A directed selection device can be used to select a lantern between each successive combination step. Such a device simply allows lanterns to be selected one at a time from one dispensing tube and to transfer the respective appropriate receiving tube (FIG. 3). FIG. 3a depicts details of a dispenser that can be used to transfer a set of objects, such as a lantern, one by one from the discharge tube to the receiving tube. Two-dimensional rotation uses position (Fig. 3b), two-dimensional planar arrangement (Fig. 3), or linear arrangement (Fig. 3d) to distribute the target set one by one from the discharge tube set to the receiving tube set. Multiple dispensers can be used in a single conversion manifold.
[0044]
However, for efficiency, a selection device such as a directed selection device can handle multiple dispensing and receiving tubes simultaneously (FIG. 4). The receiving tube can be used as a reactor and can instead be a distribution tube for distribution before the next round of synthesis. Alternatively, the lantern from each receiving tube can be transferred to the reaction vessel as long as the order is maintained. Finally, at the end of the synthesis, lanthanum can be transferred into a cleavage or storage vessel such as a 96 well plate at 1 lanthanum per well using this directed selection device. Therefore, for purely practical reasons, which conveniently accommodates the 96-well plate format, the apparatus accommodates 12 tube reactors at a time, and lanthanum is synthesized and distributed into the 96-well plate at the end of the synthesis. It can be made to allow lantern selection in between. The algorithm of the present invention allows any reasonable size synthesis of combinatorial libraries in a batch fashion with a 12 tube directed selector.
[0045]
(Chemistry)
A chemical route to the untracked synthesis of trisubstituted benzimidazoles with SynPhase lanthanum was developed according to FIG. Aminomethylated lanthanum is acylated with Emoc-4-methoxy-4- (γ-carboxypropyloxy) benzhydrylamine. Aromatic nucleophilic substitution of fluorine in o-fluoronitrobenzene with an immobilized amino group provides o-nitroaniline 1. Nitroaniline is reduced with tin (II) chloride dihydrate in N-methylpyrrolidone (NMP) and then reacted with isothiocyanate to produce thiourea 2 supported on the polymer. Cyclization to 2-arylaminobenzimidazole 3 is accomplished by diisopropylcarbodiimide (DIC). In order to introduce a third combination step, the resin-bound 2-arylaminobenzimidazole is reacted with an isocyanate to obtain the lanthanum bond product 4. Target compound 5 is obtained by cleavage from lanthanum using TFA. FIG. 6 shows the purity of the crude product.
[0046]
Directed selective combinatorial synthesis is directed through the directed selection of a support or reaction vessel in which the support or reaction vessel is physically moved, or in other embodiments they do not move, but rather the addition of reagents. This can be achieved through directed selection of the support or reaction vessel, or a combination of both. A person skilled in the art can accomplish a number of ways to control the release of the reagent to the location described by the algorithm of the present invention instead of directly selecting the support to achieve the same directed synthesis (eg, the release of the reagent in a constant reaction vessel). Can be foreseen. It is also recognized that similar functional arrays can be constructed that are physically connected by tubes and valves to produce the same functional relationship of connections between sets of reaction vessels, rather than physically to the array. One can foresee the use of an array of reaction vessels, for example a three-dimensional array (FIG. 8), in which the reaction vessels are separated from the nearest reaction vessel. For example, a reaction vessel only needs to have a single pair of valves or ports (for entry and exit of reagents, gases, etc.), but selectively access a combination of reaction vessels, for example as described by the algorithm There is a valve assembly or valve switching mechanism for the purpose.
[0047]
For illustrative purposes, an actual regular three-dimensional array of interconnected reaction vessels can be drawn, as shown for the synthesis of the 3 × 3 × 3 library of FIG. Each reaction vessel has six valves for the flow of reagents through from the valve a to the valve c, from the valve b to the valve d, and from the valve f to the valve e in the x-axis, y-axis, and z-axis directions, respectively. For illustrative purposes, the horizontal layers of the three-dimensional array of reaction vessels are x, y. z and the respective vertical layers of the three-dimensional array of reaction vessels are labeled 1,2,3. Thus, i) the opening of valves b and d allows the reagent to flow in the x-axis direction from the reaction vessel in the horizontal layer x to the vessel in the horizontal layer y to the vessel in the horizontal layer z, substantially Ii) Opening valves a and c allows the reagent to flow in the y-axis direction and re-uses an effective reaction vessel. And (iii) opening valves f and e will allow the reagent to flow in the x-axis direction and again reconfigure a valid reaction vessel.
[0048]
Therefore, three building blocks are included in each reaction (BB 1, 2, 3 in reaction 1, BB 4, 5, 6 in reaction 2 and BB 7, 8, 9 in reaction 3). Example of a synthetic reaction for a three-step reaction I) enter through open valve a and drain through open valve c of all reaction vessels in layer x (recycle if desired) to add BB1 and carry out reaction 1 , Run through open valve a and drain through open valve c of all reaction vessels in layer y (recycle if desired), add BB2, perform reaction 1 and open Enter through valve a and drain through open valve c of all reaction vessels in layer z (recycle if desired) and add BB3 to carry out reaction 1, ii) open valve f Through and open through open valve e of all reaction vessels in layer x (recycle if desired), add BB4 to carry out reaction 2 and enter through open valve f And drain through open valve e of all reaction vessels in layer y (recycle if desired) and add BB5 to perform reaction 2, enter through open valve f, and Drain through open valve e of all reaction vessels at z (recycle if desired) and add BB6 to carry out reaction 2, iii) enter through open valve b, and vertical layer Drain through open valve d of all reaction vessels in 1 (recycle if desired), add BB7 to perform reaction 3, enter through open valve b, and enter vertical layer 2 All reactions Drain through the open valve d of the vessel (recycle if desired) and add BB8 to carry out reaction 3, enter through open valve b, and all reaction vessels in vertical layer 3 Drain (through recycling if desired) through open valve d and add BB9 to carry out reaction 3. In this way all compounds are identified through opening and shutting off valves, effective reaction vessel sorting and re-partitioning, and effective selection of individual reaction vessels and the support contained within them (in the case of solid phase synthesis). Is synthesized at the position of.
[0049]
Without further consideration, it is believed that one skilled in the art can utilize the present invention to its extent using the above description. The following preferred specific embodiments are therefore to be considered merely illustrative and not limiting of the remainder of the disclosure.
[0050]
In the examples above and below, all temperatures are stated in uncorrected Celsius, and all parts and percentages are by weight unless otherwise specified.
【Example】
[0051]
Directed selective Teflon reactor:
A -28 UNF thread is cut on both ends of a Teflon tube (Cole-Palmer, Vernon Hill, IL; outer diameter 8 mm, inner diameter 5.6 mm). The length of the tube is cut to allow 5.3 mm for each lantern and 5 mm x 2 for both screws. The tube is filled with lantern and sealed from both sides using two female luer fittings with -28 UNF threads (Figure 1a). FIG. 1a depicts the use of a Teflon tube as a reaction vessel that can be capped at both ends after the target set is filled. The object set is accommodated so as to maintain the tube diameter in an ordered linear array to prevent mixing. Reagents can flow into or through the tubes, and in the case of lanterns, hollow aligned cores contain necklaces, or catalysts, etc. or serve as catalysts for handling each subject set Allow lace insertion.
[0052]
FIG. 1b shows the basic idea of the various types of reaction or discharge and receiving vessels, including lanterns, simple tubes, tubes contained in microplates, or syringes depicting its hollow core, and necklaces Describes the ability to discharge a target set into or remove a target set into such a tube. The ability to add or remove reagents is depicted as the ability to use a tube that is capped or minimally occluded enough to hold the subject set unless pressure is applied.
[0053]
Reaction on SynPhase Lanthanum:
Two lanthanum sizes are available, an L-series lantern for loading 15 mmol (5 × 5 mm size) and a D-series lantern for loading 35 mmol (5 × 12.5 m size). The concept described is applicable to either series of lanterns, but L-series lanterns are used. Both lanterns can fit into the same tube, and each has a central hole that is convenient to pass the wire and pass the lantern.
[0054]
One L-series lanthanum weighs 38 mg and gives a nominal substitution of about 0.4 mmol / g. Since lanthanum does not swell, the use of solvent is substantially lower when compared to resin beads. 67 lanterns represent 1 mmol and fill a volume of 12 mL. Lanthanum does not swell under standard conditions, so 5 mL of solvent is sufficient to wash all lanthanum. The same volume is also used to carry out each reaction. In most solid phase reactions, an excess of reagent is used, and 5 mL for lanthanum synthesis at a concentration of 0.5 M makes the reagent 2.5 mmol (2.5 molar excess). To be sufficient for synthesis to keep the same concentration with the resin, the reagent volume would have to be doubled. This is because 1 g of resin (usually loaded at 1 mmol / g) typically requires 10 mL of solvent per solvent exchange.
[0055]
Lantern cleaning:
The lanthanum-filled tube reactor is attached to the two Teflon distribution valves used to operate the Domino Blocks using 1/8 Teflon tubing. The four ports of the solvent selection valve are connected to four containers of solvent using 1/8 Teflon tubing. The common port is connected to a tube reactor selection valve. The common port of this valve was connected to the tube reactor. The second end of the tube reactor is connected to a tube reactor selection valve. The common port of this valve is connected to a vacuum drain. In order to wash the lanthanum, a suitable solvent is selected by a solvent selection valve. The flow through the tube reactor is regulated by a tube reactor selection valve. A typical volume of wash solvent is 200 mL per 50 lanthanum tube reactor.
[0056]
Support (lantern) directed selection device:
Two sorters are tested. The first uses a three-chamber principle, a support release chamber (Teflon tube), a support isolation and transfer chamber, and a support receiving chamber (Teflon tube). The discharge and receiving chambers are offset so that the support in the discharge chamber cannot pass directly into the receiving chamber. The support isolation and transfer chamber allows only one or any desired number of supports to pass from the release chamber into the isolation and transfer chamber and then these supports are transferred into this chamber or the isolation and transfer chamber It repositions itself and separates the release and reception chambers in a manner that allows the support to be transferred to the reception chamber. In the first device, the isolation and transfer chamber is incorporated into a push rod as shown in FIG. The push rod structure is a simple way to move one or a defined number of supports (regulated by the dimensions of the isolation and transfer chambers) from side to side in any desired series of steps from the discharge chamber to the receiving chamber. . Reliable transfer without clogging or transfer failure is for library synthesis, which is based on knowing the location of each support throughout the synthesis process rather than an analytical question of the label with identification of each synthesized compound. Important for the directed selection used. This reciprocating mechanism provides this reliability. Multiple reaction vessels and sorters can be used to transfer a set of objects simultaneously but individually from one set of reaction vessels to another. Multiple sorters can be arranged as a two-dimensional or three-dimensional diverting manifold. In a preferred embodiment, one such arrangement utilizes a two-dimensional circular set of sorters as a diverting manifold and a circular rotatable set of reaction or discharge vessels above a second circular set of reaction or receiving vessels. To do. If reaction vessels are used, they can be inserted. If the discharge vessel is different from the reaction vessel, a method such as the use of necklaces (rods with grippers / stops, just wires, strings, or pillars) is used for each discharge from the respective reaction vessel of the target set of columns. Can be used to transfer to a container.
[0057]
The second device is suitable for large library synthesis (FIG. 4). This directed selection device is designed to move lanterns simultaneously from 12 (or any desired number) discharge tubes (chambers) into 12 (or the same desired number) receiving vessels (chambers). Twelve distribution tubes are attached to the top of a circular stainless steel distribution manifold. The receiving tube is connected in a linear manner to the bottom of the conversion manifold. A conversion chamber containing 12 (or the same desired number) isolation and transfer chambers is connected to the circular distribution manifold by 12 tubes. The diverting manifold serves to convert the circular arrangement of distribution tubes into a linear arrangement of receiving tubes. Inside the circular distribution manifold, a spring-loaded circular section with 12 lantern-sized openings moves a single lantern from below each of the distribution tubes to a position above each of the receiving diverter tubes. When the lantern reaches the top of each receiving conversion tube, it passes through the conversion manifold into the receiving tube. The circular array of isolation and transfer chambers is designed with one isolation and transfer chamber for each of the distribution tubes and rotates by half a step from under each discharge tube to above each receiving tube. Thus, each isolation and transfer chamber directs support selection from one discharge tube to a specific receiving tube. The discharge tubes rotate on the conversion manifold so that the support from each discharge tube is discharged into all 12 receiving tubes according to any desired selection pattern. A stainless steel 2 g weight is placed on top of the lantern in all distribution tubes to ensure reliable repositioning of the lantern. Although not required, the passage of the lantern from the isolation and transfer chamber to the receiving tube is curved to translate the circular array of discharge tubes into the linear array of receiving tubes, and the receiving tube is reliable for lanthanum transfer through the curved tube. Connected to the vacuum tank via a solenoid valve to apply negative pressure or air flow to increase performance. The solenoid valve is opened for a fraction of a second as soon as the moving lantern is positioned over the receiving tube. The negative pressure gradient / air flow helps the lantern move into the receiving tube.
[0058]
One skilled in the art can appreciate that there are various ways of applying positive pressure to the top of the dispensing stack in addition to just a weight, and that positive pressure (rod or air) can be used to push the target set out of the sorter.
[0059]
FIG. 7a depicts a two-dimensional circular arrangement of the discharge tube, diverting manifold and receiving tube used for easy selection of the subject set between synthesis steps. In this embodiment, the discharge tube rotates in a circular fashion in such a manner that the discharge tube changes its position while the conversion manifold and receiving tube are stationary. Instead, the diverting manifold rotates a portion (eg half) back and forth in one step (where one step is the distance between two discharge or receiving tubes) and consists of as many chambers as there are discharge tubes, each The chamber can receive a single target set in a first position and release the target set into a receiving tube when rotated.
[0060]
FIG. 7b is a detail of the apparatus shown in FIG. 7a. The first step depicts the conversion manifold in a first position aligned with the discharge tube. Within each discharge tube is a weight that ensures that the target set enters the conversion manifold. A circular array of discharge tubes and diverting manifolds is paired with a linear array of receiving tubes (arranged in the X direction) by a flexible tube (connecting transport tube), with a vacuum at the bottom of each receiving tube . There is also a hole above the second position of the diverter manifold, to which positive pressure (atmospheric pressure or physical instrument) can be applied to push the set of objects from the diverter manifold to the receiving tube.
[0061]
Operation:
i) In the first position, the target set (from each building block labeled 1, 2, 3) for the respective discharge tube enters the conversion manifold, but is not aligned with the receiving tube Can't pass. The next operation is described between the left and right panels.
ii) The conversion manifold rotates part (half) of the step counterclockwise (viewed from above) to the second position now aligned with the respective connecting transport tube for each receiving tube A vacuum is applied transiently to the bottom of the tube, and the target set falls into each receiving tube.
iii) The conversion manifold reverses a partial step to its first position (not shown), and the target set is discharged from each reaction discharge tube into the respective conversion manifold position.
iv) The discharge tube and diverting manifold are synchronously rotated one step clockwise and are repositioned so that they are both indexed into one receiving tube away from their original position in i).
v) The process ii) hops back and the process is repeated until all target sets have been discharged into the receiving tube. At the end, the target set can be extruded from each of the respective tubes into the reaction tube (popping out to maintain the top-to-top orientation of the target set) or collected on the necklace, It can then be reacted on the necklace or transferred from the necklace into the reaction vessel. In the case of transfer from a release tube to a microplate or other receiving tube, after each ii) with one subject per well or tube (such as the end of synthesis when it is desired to release one compound per well) The receiving plate (consisting of x rows of tubes in the x linear direction) can be indexed one tube at a time over the y direction, or another set of empty tubes can be placed under the connecting transport tube Can do.
[0062]
Distribution of support into 96 well plate:
Instead of twelve receiving tubes, a 96 well plate is placed under the conversion manifold for the final distribution of lanthanum into the wells after synthesis is complete. This distribution from the column of the target set in the tube to the single target set in each well of the microplate, often in a circular or other two-dimensional arrangement, for any number of configurations of reaction vessels It will be common and will be facilitated by different configurations for multiple (conversion manifolds) in the sorter used.
[0063]
Chemistry:
Lanthanation acylation with a linker:
A 50 mL syringe is loaded with 50 lanthanum, which is neutralized with 50% piperidine in DMF and washed 5 times with DMF. Fmoc-4-methoxy-4- (γ-carboxypropyloxy) benzhydrylamine (5 mmol, 2.69 g) and HOBt. H ₂ O (5 mmol, 0.765 g) is dissolved in 15 mL of NMP and DIC (5 mmol, 0.782 mL) is added. This solution is added to the syringe holding the lanthanum and kept on the tumbler overnight (16 h). The lanthanum is washed 5 times with DMF, THF and DCM and dried by a stream of nitrogen. To quantify the amount of linker available on each lanthanum (linker substitution degree), 0.5 mL of 50% piperidine / DMF solution is added to one lanthanum in a 2.5 mL syringe and kept on the tumbler for 10 minutes. The lanthanum is washed 5 times with DMF, and the entire wash is collected, diluted and the absorbance for DMF at 302 nm is measured. Fmoc release indicates a linker substitution degree of 37 μmol / lanthanum.
[0064]
Nitroaniline: Step 1
A 20 mL syringe is charged with 10 lanthanum (acylated with a linker as described above) and a 50% piperidine / DMF solution is added. After 10 minutes, the lanthanum is washed 5 times with DMF and 3 times with dry DMSO, and 5 mL of a solution of 1-M o-fluoronitrobenzene and DIEA (0.17 mL) in DMSO is added to the syringe. The syringe is left to shake at 75 ° C. overnight (16 h) in an incubator. The lanthanum is washed 5 times with DMSO, DMF, DCM and dried with nitrogen. One ring is cut from the lanthanum and the product is cleaved with TFA for 1 hour and analyzed on an analytical gradient HPLC at 280 nm.
[0065]
o-Phenylenediamine: Step 2
A 20 mL syringe containing 10 lanthanum from step 1 is charged with 5 mL of a 2M solution of tin (II) chloride dihydrate in MMP and bubbled with argon for 15 minutes. The syringe is left on the tumbler overnight, washed 3 times with NMP, DMF, DMF / water, DMF, THF, DCM and dried with nitrogen. One ring is cut from lanthanum, the product is cleaved with TFA for 1 hour and analyzed by analytical gradient HPLC at 220 nm.
[0066]
Thiourea: Step 3
A 20 mL syringe containing 10 lanthanum from step 2 is charged with 5 mL of 1M isothiocyanate solution in NMP. The syringe is left on the tumbler overnight, washed 3 times with DMF, THF, DCM and dried with nitrogen. One ring from the lanthanum is cut, the product is cleaved with TFA for 1 hour, and analyzed by analytical gradient HPLC at 280 nm.
[0067]
Benzimidazole: Step 4
Charge a 20 mL syringe containing 10 lanterns from step 3 with 5 mL of 1M DIC solution in DMF. The syringe is left on the tumbler overnight, washed 3 times with DMF, THF, DCM and dried with nitrogen. One ring is cut from the lanthanum and the product is cleaved with TFA for 1 hour and analyzed by analytical gradient HPLC at 280 nm.
[0068]
Urea: Step 5
A 20 mL syringe containing 10 lanthanum from step 4 is charged with 5 mL of 1M isocyanate solution in DMF. The syringe is left on the tumbler overnight, washed 3 times with DMF, THF, DCM and dried with nitrogen. One ring is cut from the lanthanum and the product is cleaved with TFA for 1 hour and analyzed by analytical gradient HPLC at 280 nm.
[0069]
Virtual tube set:
As depicted in FIG. 13, multiple short tubes can be used instead of a single long tube. Each set of short tubes forms a virtual tube after aligning the head to tail as shown to the left of the vertical line. The right side of the vertical line contains each 3 object set and includes each object set on the right side of the first two steps of the synthesis of the 3 × 3 × 3 library using the short tubes that make up the virtual tube set. This is a comparison to a single long tube. The building blocks are shown as BB1, BB2, BB3 for the first reaction and as BB4, BB5, BB6 for the second reaction. Object sets are numbered so that the location within their tubes and between selections can be tracked. Initially focusing on virtual short tube synthesis, three tubes are each reacted with their own building blocks. As each 3 tube set is ordered head-to-tail, 9 subjects can be seen by comparing the number for the BB1 short tube with the long tube corresponding to the right BB1 (see dotted arrow) Represents a “virtual tube” that contains a set. Next, one tube of each reaction set BB in the tube reaction is combined with each one of the other reaction set tubes (as shown) three times in preparation for generating a three tube discharge tube set. The three tube selection is performed. The “virtual tube” head-to-tail layout dictates how these tubes are recombined before selection. Three individual selections were performed on these different sets of tubes as indicated, and the resulting receiving tube was the first versus tail “virtual tube” for reaction with the next three building blocks (BB4, BB5, BB6). Will be joined again. On the right is shown the use of 3 long tubes for the same synthesis step with a single 3 tube selection. Comparison of the head-to-tail “virtual tube” in each reaction BB4, BB5, BB6 with the long tube in the BB4, BB5, BB6 reaction (dotted arrow) shows the same configuration and order, and the tube length is convenient length Prove the use of the “virtual tube” concept with the sorter as a means to keep it (e.g., divide the 200 target set tube into four 50 target set tubes).
[0070]
All applications, patents and publications cited herein, and the corresponding US provisional application No. 1 entitled “Methods and apparatus for directed selection in combinatorial synthesis”. The entire disclosure of 60 / 307,186 is incorporated herein by reference.
[0071]
The above examples can be repeated with similar success by substituting the generally or specifically described reactants and / or reaction conditions of the present invention with those used in the above examples.
[0072]
From the foregoing description, those skilled in the art can readily ascertain the essential characteristics of the present invention, and various modifications and alterations of the present invention can be made to adapt to various uses and conditions without departing from the spirit and scope thereof. Can be made.
[Brief description of the drawings]
[0073]
FIG. 1a shows a reaction vessel, a dispensing vessel and a receiving vessel and a Teflon tube as a receiving vessel.
FIG. 1b shows a Teflon tube as a reaction vessel, a dispensing vessel and a receiving vessel.
FIG. 2 shows a selection algorithm, where each string represents a different reaction vessel, each letter represents the addition of a building block, and the numbers on each target set correspond to the order of the target set before each remix.
2a further illustrates the algorithm of FIG. 2, where each necklace represents a different reaction vessel and each letter represents the addition of a building block. The relationship of building blocks after each remixing and after each building block is added is shown.
FIG. 3a depicts details of a dispenser that can be used to transfer a set of objects, such as a lantern, one by one from a discharge tube to a selected receiving tube.
FIG. 3b depicts an arrangement using multiple dispensers in a conventional diversion manifold to distribute a set of objects one by one from a set of discharge tubes to a set of receiving tubes using a two-dimensional rotational arrangement.
FIG. 3c depicts an arrangement using multiple dispensers in a conventional diverter manifold to distribute a set of objects one by one from a set of discharge tubes to a set of receiving tubes using a two-dimensional planar arrangement.
FIG. 3d depicts an arrangement using multiple dispensers in a conventional diverter manifold to distribute a set of objects one by one from a set of discharge tubes to a set of receiving tubes using a linear arrangement.
FIG. 4 illustrates a 12-channel reshuffler.
FIG. 5 illustrates the untracked synthesis of benzimidazole.
FIG. 6 is an analytical gradient HPLC profile of crude benzimidazole.
FIG. 7 depicts a linear arrangement of discharge tube, diverting manifold, and receiving tube that is used to easily select a target set between synthesis steps.
7b is a detail of the apparatus shown in FIG.
FIG. 8 is a three-dimensional array in which each reaction vessel is separated from the nearest adjacent reaction vessel by a valve.
FIG. 9 is a diagram of a 3 building block × 4 building block library.
FIG. 10 is a diagram of a 4 building block × 3 building block library.
FIG. 11 depicts a three step process using three building blocks at each step. Each string is a separate reaction vessel with its own 9 support.
12 depicts the three step process of FIG. The building block relationship is shown.
FIG. 13 depicts the use of multiple short tubes to form a virtual tube instead of a single long tube.

Claims (57)

A method of selecting a target set into a group for combinatorial compound synthesis by sequential addition of building block molecules comprising:
a) redistributing individual object sets from a first group of object sets in a fixed first order to a second group of object sets, wherein the object sets are in a different second order; Redistributed, so that after redistribution, there are no adjacent target sets from the first ordered group of target sets that are adjacent in the second ordered group of target sets;
b) after said redistribution, adding a chemical building block to each group of subject sets; and c) repeating steps a) and b) to add each desired building block;
Including methods. The method of claim 1, wherein the object sets are contained in at least one holder to maintain their order during handling. The method of claim 2 wherein the number of holders is equal to the number of building block reactions for that step. The method of claim 2, wherein the holders are grouped into a linear set for each building block reaction, such that the linear set forms a virtual holder. The method of claim 2, wherein the object set is inserted into a necklace. 3. The method of claim 2, wherein the target set is removed using a necklace. The method of claim 1, wherein the object sets are included on the necklace to maintain their order during handling. The method of claim 2 wherein the holder is open at both ends. 9. The method of claim 8, wherein the holder can be squeezed to hold the object set. 9. The method of claim 8, wherein the object set can be inserted through one end of the holder. 9. The method of claim 8, wherein the object set can be removed from one end of the holder. The method of claim 2 wherein the holder is open at one end. The method of claim 1, wherein the same order of redistribution is repeated at each step of the synthesis. The method of claim 1, wherein the same order of redistribution is repeated in all but one step of the synthesis. 15. The method of claim 14, wherein the redistribution order for the last synthesis reaction step is different. 15. The method of claim 14, wherein the distribution order prior to the second reaction step is different. The method of claim 1, wherein the number of subject sets in each step is equal to the total number of compounds synthesized. The method of claim 1, wherein each set of objects can be located individually, and the history of building block additions can be identified at any point in time. The method of claim 1, wherein each set of objects is greater than one. The method of claim 4, wherein all members of a set of subjects at a given time are reacted with the same chemical building block throughout the synthesis. The method of claim 1, wherein one subject set comprises one or more reaction vessels or solid supports. The method of claim 1, wherein the object set itself moves physically between groups. The target set is redistributed to any group without actually moving itself, reflecting its previous and planned chemical building block additions, and the combination of accessibility of the reagents to each target set is Always defined and described by the position of the target set, and the distribution of the target set follows the formula Nx = NtotBBx target set, where Ntot = total number of compounds synthesized Nx = target set in the reaction vessel in the xth combination step The method of claim 1, wherein BBx = total number of building blocks in the xth reaction step. The order of redistribution of target sets allows access to each target set in the group in such a way that access to each target set in each building block reflects its previous and planned chemical building block additions in any group. 24. The method of claim 23, wherein the combination of accessibility of the reagents to each target set is defined and described by the location of the target set at any time. A directed selection device for performing solid phase combinatorial synthesis of compounds comprising:
a) one or more first reaction vessels for holding one or more target sets organized in a first order pattern;
b) one or more isolation and transfer chambers for directed redistribution of the target set from each of the reaction vessels;
c) including one or more subsequent reaction vessels for receiving a redistributed subject set from said isolation and transfer chamber;
Here, the redistribution pattern of the target set in each subsequent transfer is the same, and the position of each target set in one reaction vessel encodes its previous synthesis history, and the target set is one from each reaction vessel. The target set is transferred into a subsequent reaction vessel using an iterative process in which each subsequent reaction vessel includes an Nx target set, where Nx = Ntot / BBx target set;
Ntot = Number of target sets in the xth reaction step BBx = Directed selection device that is the number of building blocks in the xth reaction step. 26. The directed selection device of claim 25, wherein the single object set is an individual object or a number thereof, and each object set maintains adjacency through all redistribution and building block additions. 26. The directed selection device of claim 25, wherein all members of a single subject set are reacted with the same chemical building block throughout synthesis. 26. The directed selection device of claim 25, wherein the single target set is one or more reaction vessels or solid supports. 26. The apparatus of claim 25 for performing a combined solid phase synthesis of compounds comprising:
a) performing the reaction in one or more reaction vessels for holding one or more sets of objects organized in a one-dimensional linear array;
b) discharging the target set from each of the reaction vessels into an isolation and transfer chamber for directed redistribution;
c) discharging the target set into one or more subsequent reaction vessels for receipt from the isolation and transfer chamber, and d) after each redistribution of the target set that remains adjacent in the second group of target sets. A directed selection device comprising repeating steps a), b), c) for each building block added so that there are no adjacent target sets from the first group. 30. The directed selection device of claim 29, wherein the single object set is an individual object or a number thereof and each of the object sets maintains adjacency through all redistribution and building block additions. 30. The directed selection device of claim 29, wherein all members of a single subject set are reacted with the same chemical building block throughout synthesis. 30. The directed selection device of claim 29, wherein the single target set is one or more reaction vessels or solid supports. The method of claim 2, wherein each target set is individually selected by an isolation and transfer chamber from the discharge holder to the receiving holder. 34. The method of claim 33, wherein the discharge holder and receiving holder are offset so that the object set cannot pass between them without a distribution device. A directed selection device for performing combined solid-phase synthesis of compounds comprising:
a) one or more first reaction vessels for holding one or more target sets contained in their respective holders and organized in a first order pattern;
b) one or more isolation and transfer chambers for directed redistribution of the target set from each of the reaction vessels;
c) one or more subsequent reaction vessels for receiving a set of redistributed objects from said isolation and transfer chamber;
d) repeating steps a), b), c) for each building block to be added,
Here, the redistribution pattern of the target set in each subsequent transfer is the same, and the position of each target set in one reaction vessel encodes its previous synthesis history, and the target set is one from each reaction vessel. The target set is transferred into subsequent reaction vessels using an iterative process to transfer each subsequent reaction vessel until it contains an Nx target set, where Nx = Ntot / BBx target set;
Ntot = Number of target sets in the xth reaction step BBx = Directed selection device that is the number of building blocks in the xth reaction step. 36. The apparatus of claim 35, wherein the discharge holder and receiving holder are offset so that the object set cannot pass between them without the distribution device. 36. The apparatus of claim 35, wherein the isolation and transfer chamber receives a single object set from the discharge holder at a time, moves from the discharge holder position to the receiving holder position, and discharges the object set to the receiving holder. 34. The method of claim 33, wherein a plurality of discharge holders and receiving holders are used. 34. The method of claim 33, wherein a plurality of isolation and transfer chambers are used to transfer a set of objects from a plurality of discharge containers to a plurality of receiving containers. The method of claim 1, wherein each of the discharge containers accommodates one or more target sets. The method of claim 1, wherein each receiving container contains one or more target sets. The method of claim 1, wherein each of the receiving containers contains a single set of objects. The method of claim 2, wherein each object set is individually selected from a necklace into one receiving holder. The target redistribution pattern in at least one (or two and all) transfers is the same, and the position of each target set in one reaction vessel encodes its previous chemical history, and the target set is the next reaction Each container is transported one at a time into the next reaction container until it contains the Nx target set, where Nx = Ntot / BBx target set,
2. The method of claim 1, wherein Ntot = total number of compounds to be synthesized Nx = number of target sets in the reaction vessel in the xth combination step BBx = number of building blocks in the xth reaction step. 45. The method of claim 44, wherein after each redistribution, there are no adjacent object sets from the first group of object sets in the release container that continue to be adjacent to the second group of objects in the receiving container. The method of claim 1, wherein the plurality of reaction vessels are in the form of microplates. The method of claim 1, wherein the reaction vessel is a syringe, a Teflon tube, a closed end tube, or an open end tube. The method of claim 1, wherein the object sets are ordered in a manner that the holes in each object set are aligned to form a hollow passage through all object sets in the holder. The method of claim 2 wherein the solution flows through and around all object sets in the holder. The method of claim 1, wherein the object set to be transferred is from the end of the stack of object sets. The method of claim 1, wherein the object set moves through the dispensing tube to the transfer device. 40. The method of claim 39, wherein a positive pressure or weight on the top of the column of the subject set ensures active movement. 43. The method of claim 42, wherein a plurality of positive pressure devices are connected to a plurality of distribution tubes. 42. The method of claim 41, wherein negative pressure is used to move the object set into the repositioning device. The method of claim 1, wherein the object set moves from the transfer device into the receiving tube. 45. The method of claim 44, wherein positive pressure is used to move the object set from the repositioning device into the receiving tube. 45. The method of claim 44, wherein a negative pressure applied to the receiving tube is used to move the correspondence set from the repositioning device into the receiving tube.

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