WO2001019484A9 - Dispositif et procede de purification - Google Patents

Dispositif et procede de purification

Info

Publication number
WO2001019484A9
WO2001019484A9 PCT/US2000/024978 US0024978W WO0119484A9 WO 2001019484 A9 WO2001019484 A9 WO 2001019484A9 US 0024978 W US0024978 W US 0024978W WO 0119484 A9 WO0119484 A9 WO 0119484A9
Authority
WO
WIPO (PCT)
Prior art keywords
target compound
quenching reagent
residue
reagent
quenching
Prior art date
Application number
PCT/US2000/024978
Other languages
English (en)
Other versions
WO2001019484A1 (fr
Inventor
Shahnaz G Jamalabadi
Original Assignee
Dyax Corp
Biotage Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dyax Corp, Biotage Inc filed Critical Dyax Corp
Priority to AU75777/00A priority Critical patent/AU7577700A/en
Priority to EP00964973A priority patent/EP1265684A4/fr
Priority to CA002388747A priority patent/CA2388747A1/fr
Priority to JP2001523106A priority patent/JP2003509185A/ja
Publication of WO2001019484A1 publication Critical patent/WO2001019484A1/fr
Publication of WO2001019484A9 publication Critical patent/WO2001019484A9/fr

Links

Classifications

    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
    • B01J20/3204—Inorganic carriers, supports or substrates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
    • B01J20/289—Phases chemically bonded to a substrate, e.g. to silica or to polymers bonded via a spacer
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
    • B01J20/3206—Organic carriers, supports or substrates
    • B01J20/3208—Polymeric carriers, supports or substrates
    • B01J20/321—Polymeric carriers, supports or substrates consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3214—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the method for obtaining this coating or impregnating
    • B01J20/3217—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
    • B01J20/3219—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond involving a particular spacer or linking group, e.g. for attaching an active group
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3244—Non-macromolecular compounds
    • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
    • B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3244—Non-macromolecular compounds
    • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
    • B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
    • B01J20/3251—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
    • B01J20/3244—Non-macromolecular compounds
    • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
    • B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
    • B01J20/3255—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
    • B01J20/3289—Coatings involving more than one layer of same or different nature
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes

Definitions

  • the invention relates to a purification device for use in chemical synthesis and method of using the device.
  • peptide, DNA, and small organic molecules libraries can be constructed using either solid phase or solution phase synthetic methodologies.
  • solution phase synthesis could produce higher purity and lower cost final product than solid phase synthesis.
  • solution phase synthesis can lead to the formation of an emulsion which can lower the purity and yield of the final product and variability of results.
  • the invention features a method of removing a residue from a target compound mixture.
  • the method includes contacting a target compound mixture including a target compound and a residue with a quenching reagent, flowing the target compound mixture across the quenching reagent, immobilizing the residue on the quenching reagent by forming a covalent bond between the residue and the quenching reagent, and removing the target compound from the quenching reagent.
  • the quenching reagent is coupled to an insoluble support.
  • the invention features a method of deprotecting a target compound having a protecting group residue.
  • the protecting group residue is an acid labile group or a base labile group.
  • the method includes the steps of flowing the target compound mixture across a quenching reagent coupled to an insoluble support to allow the covalent binding of the protecting group residue to the quenching reagent, and removing the target compound from the quenching reagent.
  • the protecting group residue can be a fluorenyl derivative.
  • the quencing reagent includes a piperazino, a piperidino, or functionally equivalent moiety.
  • the quenching reagent can be coupled to the soluble support by a linker, e.g., a linear or aromatic linkers, e.g., a linker described herein.
  • a linker e.g., a linear or aromatic linkers, e.g., a linker described herein.
  • the insoluble support can be an organic polymer or an inorganic oxide such as silica.
  • the method can include the steps of introducing the target compound mixture into a housing including the quenching reagent, and removing the target compound from the housing.
  • Flowing can include moving a solvent through the housing or cartridge.
  • the invention features a device for purifying a target 0 compound.
  • the device includes a first opening and a second opening. The first opening and the second opening are connected by a fluid flow path.
  • the device also includes a quenching reagent immobilized on an insoluble support contained within the fluid flow path.
  • the quenching reagent is capable of forming a covalent bond with a residue.
  • the residue can be a protecting group residue and the protecting group residue 5 can be an acid labile group or a base labile group.
  • the device can also include a second quenching reagent immobilized on a second insoluble support.
  • the residue can be a protecting group or an excess reagent.
  • the residue and the target compound can be covalently bonded, such as when the residue is a protecting group.
  • the method can also include the step of o releasing the residue from the target compound.
  • the insoluble support can be an organic polymer or an inorganic oxide such as silica.
  • the housing can include a cartridge.
  • the cartridge can include a solid phase support.
  • the quenching reagent can include a nucleophilic or an electrophilic group or a base, such as an amine, an isocyanate, an aldehyde, or an acyl halide.
  • the quenching reagent can include a polystyrene functionalized with the quenching reagent or a silane functionalized with the quenching reagent.
  • the target compound can be a polypeptide, a nucleotide, or an organic o compound.
  • a polypetide can include, for example, 2 or more natural or unnatural amino acids.
  • Nucleotides include, but are not limited to, any sugar-phosphate-based moiety, as well as any derivatized sugar-phosphate-based moiety.
  • organic compounds include but are not limited to, organic molecules containing a functional group protected by an acid or base labile group which reacts with the quenching agent 5 to form a covalent bond.
  • the polymer-supported quenching reagents are functionalized polymer resins.
  • the functional groups on the resins selectively bind to the residue via a covalent bond linkage.
  • Solution phase synthesis can offer certain advantages over solid phase synthesis. For example, a vast number of solution phase reactions have been developed in the synthesis literature, whereas relatively fewer reactions have been optimized for solid phase synthesis. The variety of protecting groups that can be used in solution 5 phase synthesis overwhelms the relatively limited number of solid-phase synthesis resins. Solution phase synthesis can also lead to the elimination of resin attachment and cleavage steps during synthesis. In addition, solution phase synthesis is amenable to monitoring by ordinary chromatographic or spectroscopic techniques. Solid phase reactions can be more challenging to monitor.
  • Polymer-supported quenching reagents have improved the purity and reproducability of solution phase synthesis.
  • polymer supported reagents can be used as a rapid purification technique in solution-phase parallel synthesis.
  • by-products or solution-phase reagents can be selectively trapped on a resin.
  • Previously used purification methods is such as crystallization, extraction, and chromatography can be difficult to apply when synthesizing a diverse library of compounds.
  • separating a structurally diverse array of compounds based on physical properties such as solubility or partition coefficient can present a daunting challenge when preparing libraries of compounds.
  • the use of polymer-supported quenching reagents as a purification platform allows
  • Flowing a reaction mixture across a supported quenching reagent can overcome some of the difficulties encountered in solution phase method of treating a reaction mixture with a functionalized resin in a batch context. For example, time
  • 25 usage can be more efficient.
  • the general procedure for removing the protecting group and purifying the solution phase product with polymer supported quenching reagents can requires between 14 hours and 4 days in a batch process. This reaction duration is long for combinatorial synthesis and purification situations.
  • the reaction speed is significantly increased by flowing the target compound mixture across the quenching
  • a batch process can require the use of a large excess of polymer resin.
  • typical solution phase methods require 1 mole of polymer resin per 35 0.2 mole of blocked product, significantly increasing the cost of the deblocking (e.g., deprotecting) step and the purification of the final product.
  • deblocking e.g., deprotecting
  • purification of the final product By flowing the target compound mixture across the quenching reagent coupled to an insoluble support, less resin can be used to achieve the same or improved purity of product.
  • batch processing can result in lower reaction yields due to transfer losses and material losses during filtration of the resin.
  • Multiple transfers of the product and filtration result in loss of product. This can be particularly problematic s if the loss occurs near the end of a multiple step synthesis.
  • Flowing the target compound mixture across the quenching reagent coupled to the insoluble support increases the efficiency of the reaction and the purity of the product, thereby increasing reaction yields and purity.
  • FIG. 1 is a schematic diagram depicting a device and method for purifying a target compound mixture having an excess of a reagent.
  • FIG. 2 is a schematic diagram depicting a device and method for purifying a compound.
  • FIG. 3 is a schematic diagram depicting a device and method for purifying a target compound having a protecting group.
  • Fig. 4 is structural representation of several of reagents coupled to an o insoluble support.
  • Fig. 5 is a diagram of reactions involved in Bsmoc removal.
  • the purification device includes a quenching reagent coupled to an insoluble support.
  • the quenching reagent can be covalently bonded to a polymeric support material.
  • the quenching agent can be covalently bonded to an inorganic support material such as silica.
  • a target compound which is a product of a synthetic reaction, can be separated from unwanted residues in a target compound mixture by selectively forming a covalent bond between the residue and the quenching reagent.
  • the residue can be excess starting material or a protecting group that has been removed from the target compound.
  • the residue reacts with the quenching reagent to form the 5 bond. This reaction immobilizes the residue on the insoluble support. Removal of the residue in this manner purifies the target compound once the target compound is removed from the quenching reagent coupled to the insoluble support.
  • the target compound mixture By flowing the target compound mixture across the quenching reagent, the target compound mixture is exposed to a continual supply of unreacted quenching reagent.
  • the effective increase concentration of the unreacted quenching reagent increases the kinetics of the reaction between the residue and the quenching reagent.
  • the contact time between the target compound mixture and quenching reagent is increased. This arrangement can allow the reaction to proceed more rapidly and more completely.
  • Coupling of the quenching reagent to the insoluble support permits the supported quenching reagent to be contained within a device, such as a disposable column. The device allows the target compound mixture to be passed through the column. Because the quenching reagent is coupled to the insoluble support which in turn is contained within the device, the target compound mixture can flow across the quenching reagent. Accordingly, the purification of the target compound can proceed in higher yield in a shorter amount of time.
  • the quenching reagent can be a functionality that can react to form a covalent bond with an electrophile (e.g., an amine which can react with an acyl halide or other electrophilic residue).
  • an electrophile e.g., an amine which can react with an acyl halide or other electrophilic residue
  • an nucleophile e.g., an isocyante which can react with an amine or other nucleophilic residue.
  • the reagent-coupled insoluble support can be mixed with other solid support material.
  • the other solid support material can be ordinary chromatography column packing material, such as organic polymer supports or inorganic oxide supports. Suitable support materials can include cross-linked polystyrene beads, silica, or reverse-phase silica.
  • the quenching reagent can be coupled by reaction with the surface functionality of the insoluble support.
  • a commercially available an amine quenching reagent-coupled resin is Amberlyst 15 (Aldrich Chemical Co.).
  • Quenching reagents e.g., amines and isocyanates
  • Quenching reagents can be coupled to a polymeric support as described, for example, in Booth and Hodges, J. Amer. Chem. Soc. 119:4882-4886 (1997) or in Knapczyk et al. J. Org. Chem. 48:661-665 (1983), each of which is incorporated herein by reference.
  • Quenching reagents can be coupled to inorganic supports, such as silica, as described in Carpino et al. J. Org. Chem. 48:666-669 (1983), each of which is incorporated herein by reference.
  • the synthesis of a target compound can be facilitated by using reagents coupled to an insoluble support.
  • the reagent is a quenching reagent that can, for example, form a covalent bond with a protecting group of the target compound
  • the protecting group can be immobilized on the insoluble support.
  • the reaction can be facilitated by exposing the compound to an excess amount of the coupled reagent.
  • One way to facilitate the reaction is to flow a solution of the compound over the support containing the immobilized reagent.
  • a reagent coupled to an insoluble support can be packed in a disposable cartridge and the target compound can be exposed to the reagent by flowing a solution of the compound through the cartridge.
  • the reaction can purify the target compound by removing the protecting group residue from the solution.
  • the insoluble support can be a resin or other chromatographic material, such as silica.
  • the solution containing the target compound is a liquid phase.
  • the insoluble support is a solid or stationary phase.
  • Adsorption is the equilibrium distribution of target compound or the protecting group 0 between stationary phase and the liquid phase.
  • the adsorption capacity of the insoluble support or resin can be an important parameter to control to achieve adequate purification.
  • An excess of a starting material can be used in a synthetic reaction to drive the reaction to completion without fear of complicating isolation and purification of the s final products by using a column including a coupled quenching reagent.
  • reagent A e.g., an isocyanate compound or acyl halide
  • reagent B e.g., an amine compound
  • the excess reagent is effectively removed by flowing the target compound mixture, which includes target compound 10 and excess residue 20, through device 25.
  • o Device 25 can be a cartridge or column used in chromatography.
  • Device 25 includes housing 30.
  • Housing 30 has first opening 40 and second opening 50, connected by fluid flow path 55.
  • Fluid flow path 55 within housing 30, contains insoluble support 60.
  • Insoluble support 60 is coupled to quenching reagent 70 (e.g., an amine).
  • Quenching reagent 70 is capable of forming a covalent bond with 5 excess residue 20.
  • the quenching reagent can be an electrophile (e.g., an isocyanate) which can form a covalent bond with an unreacted amine.
  • a target compound 10 bearing protecting group residue 20 can also be purified by the method.
  • Protecting group residue 20 is base labile (i.e., 5 is removed under basic conditions) and reacts to form a covalent bond with an amine.
  • the protected compound can be purified by passing the target compound mixture, which includes protecting group residue 20 covalently bonded to target compound 10, though device 25, including housing 30, and first opening 40 and second opening 50, connected by fluid flow path 55.
  • fluid flow path 55 within housing 30, contains insoluble support 60.
  • Insoluble support 60 is coupled to quenching reagent 70 (e.g., an amine).
  • Device 25 can also include solid phase support 5 65 (e.g., other chromatography beads). Quenching reagent 70 is capable of forming covalent bond 80 with protecting group residue 20.
  • a fluorenylmethoxycarbonyl (Fmoc)-protected amino acid was deprotected and purified using the device including a piperazine coupled to a silica support prepared as described Carpino et al. J. Org. Chem. 48:666-669 (1983).
  • 200 mg 0 of Fmoc-Ile-DCPM in 1 mL of DMF was injected into a Flash 12 cartridge (Biotage) packed with the silica-supported piperazine.
  • the cartridge was washed with 10 mL of DMF at a flow rate of 0.4 mL per minute for an elapsed wash time of 250 minutes.
  • an alkylating reagent e.g., an epoxide
  • an electrophilic quenching reagent such as an isocyanate, o which is coupled to an insoluble support in the device, can be used to purify the product by immobilizing the excess secondary amine on the quenching reagent.
  • an electrophilic scavenger device and a nucleophilic scavenger device 5 can be used sequentially.
  • a mixed bed device can be used.
  • two or more layers of quenching reagent functionalized insoluble support can be used.
  • the device can have two layers.
  • One layer can include an insoluble support having a quenching reagent capable of forming a covalent bond with a nucleophile.
  • a second layer can be placed above or below the first layer.
  • the second o layer can include a coupled quenching reagent that is capable of forming a covalent bond with an electrophile.
  • a solid phase support layer that is not functionalized can be placed in between the first layer and the second layer to prevent adverse interactions between the layers.
  • the order of exposure each layer to the target compound mixture can depend on the nature of the residues in the target compound mixture. The order of 5 exposure that provides the target compound in highest purity can be more efficient to use in the purification method, although this must be determined for each type of target compound mixture produced.
  • Scheme I depicts a series of synthetic reactions that can be carried out under excess reagent conditions to provide a target compound mixture that can be purified by the method using an electrophilic quenching reagent device, a nucleophilic quenching reagent device, or a mixed layer device.
  • silica-base reagents Four non-swelling silica-base reagents were prepared and compared for carrying out de-blocking/scavenging reactions by continuous methods, e.g., column, and batch 5 technique. Silica reagents bearing the piperazino and piperidino function were prepared. These organic bases were attached to silica using linear and aromatic linkers. Fig. 4 shows the four linker/reagent combinations.
  • Piperazino and piperidino functionalized silica gel were synthesized and evaluated in 0 batch and on-column procedures. For on-column use, disposable columns (Biotage Flash Samplets) were prepared. These products were designed to: (1) de-block the amino protecting group 9-fluorenylmethyloxycarbonyl (Fmoc); and (2) scavenge the dibenzofulyene liberated in the de-blocking process. The application of these functionalized silica reagents was also shown for l,l-Dioxobenzo[b]thiophene-2- 5 ylmethyloxycarbonyl (Bsmoc) amine protecting group.
  • Bsmoc 1, l-Dioxobenzo[b]thiophene-2-ylmethyloxcarbonyl
  • Bsmoc is a common base sensitive amino group protecting group. See Fig. 5.
  • Bsmoc undergoes Michael-like 0 addition by base to generate the free amine and the reactive intermediate (2).
  • the intermediate (2) decays over 8-10 minutes to give the stable de-blocking product (3).
  • Bsmoc- / -chlorocarbanilate (Bsmoc-PCA) was used as test probe, lg of piprazinobenzyl-functionalized silica (reagent 2) was packed in the samplet. 30mg, 60mg, 100 mg, or 130 mg of Bsmoc-PCA dissolved in 2 ml of DMSO placed on samplet. The solution was left in the samplet for 30 minutes. De-blocking was 100% o complete for the 30 and 60 mg of samplet. 98% de-blocking was achieved with the 100 mg of sample. Bsmoc-de-blocking is dependent on, time, sample load, and solvent composition.
  • Piperazino-benzyl-functionalized silica (Reagent 2) was introduced for on-column de- 5 blocking and scavenging of Fmoc and Bsmoc groups.
  • This reagent is very successful in Biotage samplet format and allows the automation of parallel solution phase synthesis using Fmoc-chemistry.
  • Base functionalized silica gel can be used in different applications including: synthesis of peptide using Fmoc- and Bsmoc- chemistry; solid phase organic synthesis; solid phase Knoevenagel catalyst. Unreacted excess o starting material is selectively removed from reaction mixture.
  • solid supported de-blocking reagents include: no loss of product due to formation of emulsion; eliminate extractions; decrease the quantity of solvents utilized; decrease the number of steps in the process including no filtration of solid support; and 5 allow automation of parallel solution phase chemistry.

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Inorganic Chemistry (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)
  • Saccharide Compounds (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne un dispositif et un procédé de purification. Le dispositif (25) comprend un réactif (70) de désactivation couplé à un support insoluble (60). Un mélange de composé cible (10) traverse ledit réactif (70), immobilisant les résidus d'impuretés (20) sur ledit réactif (70) et purifiant le composé cible (10).
PCT/US2000/024978 1999-09-13 2000-09-13 Dispositif et procede de purification WO2001019484A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU75777/00A AU7577700A (en) 1999-09-13 2000-09-13 Purification device and purification method
EP00964973A EP1265684A4 (fr) 1999-09-13 2000-09-13 Dispositif et procede de purification
CA002388747A CA2388747A1 (fr) 1999-09-13 2000-09-13 Dispositif et procede de purification
JP2001523106A JP2003509185A (ja) 1999-09-13 2000-09-13 精製装置および精製方法

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US7094351B2 (en) 2000-10-12 2006-08-22 Corcoran Robert C Purification of substances by reaction affinity chromatography
BR0308577A (pt) * 2002-03-20 2005-01-04 Pharmacia Italia Spa Reagentes suportados por polìmero para purificação de produtos naturais
US6820506B2 (en) 2002-03-27 2004-11-23 3M Innovative Properties Company Multi-chambered pump-valve device
GB0410448D0 (en) * 2004-05-11 2004-06-16 Hammersmith Imanet Ltd Purification methods
EP2845608A1 (fr) 2013-09-09 2015-03-11 Trasis S.A. Procédé de purification d'analogues de choline étiquetés 18F
WO2023100780A1 (fr) * 2021-11-30 2023-06-08 三井金属鉱業株式会社 Cartouche de matériau adsorbant et colonne de traitement de liquide l'utilisant

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US5405938A (en) * 1989-12-20 1995-04-11 Anti-Gene Development Group Sequence-specific binding polymers for duplex nucleic acids
CA1315968C (fr) * 1987-04-24 1993-04-13 David Colin Sherrington Couche sous-jacente et methode de fabrication de ladite couche
US5049656A (en) * 1988-12-21 1991-09-17 Board Of Regents Of The University Of Nebraska Sequential peptide and oligonucleotide syntheses using immunoaffinity techniques
JP2960257B2 (ja) * 1992-06-04 1999-10-06 ピーイーバイオシステムズジャパン株式会社 ビオチン導入試薬およびそれを用いる合成ペプチド精製法
DE69404848T2 (de) * 1993-08-13 1998-03-19 Minnesota Mining & Mfg Filterpatrone mit darauf angeordneten unlöslichen enzympartikeln
US5576453A (en) * 1994-07-18 1996-11-19 Es Industries Stable, sterically-protected, bonded phase silicas prepared from bifunctional silanes
WO1997042230A1 (fr) * 1996-05-03 1997-11-13 Warner-Lambert Company Purification rapide par reactifs de refroidissement brusque supportes par des polymeres
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CA2388747A1 (fr) 2001-03-22
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AU7577700A (en) 2001-04-17
EP1265684A1 (fr) 2002-12-18
EP1265684A4 (fr) 2003-04-16

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