WO2006126760A1 - Resine a structure coeur-ecorce ('core-shell') de type gel - Google Patents

Resine a structure coeur-ecorce ('core-shell') de type gel Download PDF

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WO2006126760A1
WO2006126760A1 PCT/KR2005/003273 KR2005003273W WO2006126760A1 WO 2006126760 A1 WO2006126760 A1 WO 2006126760A1 KR 2005003273 W KR2005003273 W KR 2005003273W WO 2006126760 A1 WO2006126760 A1 WO 2006126760A1
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resin
shell
core
linker
amino methyl
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PCT/KR2005/003273
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English (en)
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Yoonsik Lee
Hanyoung Kim
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Seoul National University Industry Foundation
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Publication of WO2006126760A1 publication Critical patent/WO2006126760A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/12—Powdering or granulating
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • C08J3/245—Differential crosslinking of one polymer with one crosslinking type, e.g. surface crosslinking
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F8/00—Chemical modification by after-treatment
    • C08F8/30—Introducing nitrogen atoms or nitrogen-containing groups
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • C08J3/26—Crosslinking, e.g. vulcanising, of macromolecules of latex
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2325/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an aromatic carbocyclic ring; Derivatives of such polymers
    • C08J2325/02—Homopolymers or copolymers of hydrocarbons
    • C08J2325/04—Homopolymers or copolymers of styrene
    • C08J2325/06—Polystyrene

Definitions

  • the present invention relates to a gel type of core-shell resin. More particularly, the present invention is directed to a gel type core-shell resin composed of a core section in which amino methyl ends of the amino methyl polystyrene resin cross-linked with divinyl benzene, are cross-linked with cya ⁇ tiric chloride; and a shell section made up of polyalkylene glycol resin terminated with hydroxy group or amino group at both ends and combined at the remaining chloride end of the triazine participating in the crosslink between the above amino methyl ends.
  • the combinatorial chemistry means a technology employed for the efficient preparation of lots of compound libraries by means of various combination between compounds.
  • a method used for preparation of lots of compound libraries at once through combination is referred to as combinatorial synthesis.
  • the reason that the solid-phase synthesis is preferred to the liquid-phase peptide synthesis, is that the excessive reactants and by-products which are not linked to solid-phase can be removed during a washing process since the target material is linked to polymeric support, and also the reaction can be completed by the use of excessive substrates since the substrates used excessively may be removed from solid-phase supporter by washing with unreactive and volatile solvent after the completion of reaction. Therefore, recently the research on the solid phase synthesis has been proceeded actively since, in general, the solid phase synthesis is suitable for combinatorial chemistry field.
  • the technology employed for the solid phase synthesis consists of three essential elements.
  • the first is the polymeric support
  • the second is a linker that links a polymeric support with a scaffold or a target compound
  • the third is the scaffold or the target compound.
  • the polymeric support should be stable when it is used with various organic solvent and reagent, and in various conditions of reaction.
  • the most widely used polymeric support is gel type resin cross-linked to the extent of 1 % to 2%.
  • polystyrene resin was used primarily since its mechanical strength is good and functional groups are easily grafted on polystyrene.
  • the polystyrene resin has several problems that enzyme reaction or oligonucleotide synthesis which should be performed in hydrophilic media, increases as the application field of solid-phase reaction increases, that a compatibility between hydrophobic polystyrene resin and a hydrophilic peptide chain of high molecular peptide decreases, and that the efficiency of coupling reaction falls off as the peptide chain grows longer since nonpolar hydrophobic solvent, such as CH 2 Cl 2 used for swelling the polystyrene resin, is not able to easily dissolve the peptide chain.
  • nonpolar hydrophobic solvent such as CH 2 Cl 2 used for swelling the polystyrene resin
  • PEG has biosusceptibility and minimize the adsorption of protein
  • the PEG grafted polystyrene resin may decrease steric hindrance effect to a certain degree similarly with that PEG is introduced on polymer support as a linker and the coupling yield can be improved by increasing the compatibility between peptide chain and the polymeric support by means of the hydrophilicity of PEG.
  • the salt genera t ed in the course of peptide synthesis may be removed more effectively since the swelling properties of the PEG grafted gel type resin increases even for high polarity solvent
  • TentaGel, BT-GeI 1 and the like used widely recently, are the typical examples of the resin in which PEG is grafted on cross-linked polystyrene,
  • cross-linked polystyrene-polyethylene glycol grafted resin forms a homogeneous phase all over and is not separated into polystyrene domain and polyethylene glycol domain,
  • a cross-linked polystyrene resin of polymeric support forms a structure that makes polymeric support not be dissolved but be swelled in reaction solvent, the polyethylene glycol resin grafted on the polystyrene cross-linked structure, complements the hydrophobicity of the polystyrene resin, and has miscibility with nucleotide, reacion product, or amino acid participating in synthetic reaction for polypeptide or for oligonucleotide,
  • the synthetic reaction of polypeptide or of oligonucleotide is proceeded at the end of the polyethylene glycol chain which exists homogeneously overall of the support resin.
  • the polystyrene structure blended with the PEG inhibits the diffuse and penetration of amino acid or nucleotide into the polymer support.
  • the synthetic reaction is proceeded in the bulk phase of the polymer support as well as in the surface of the polymeric support resin, in case of the solid phase synthesis by using the conventional polymer support.
  • it is difficult to control the progress of the reaction or the extent of the reaction in the bulk phase of the polymer support, since it is not easy for amino acid or nucleotide to diffuse and penetrate info a support resin due to the existing polystyrene resin.
  • the reaction yield is also low, since it is difficult to separate and recover the reaction product synthesized in the bulk phase of the polymer support.
  • it is not desirable for large-scale synthetic process for peptide since the substitute rate of the functional group decreases to ⁇ 0.2mmol/g due to the graft of polyethylene glycol chain.
  • the polystyrene resin composed of crossli ⁇ ked structure of the conventional polymeric support resin does not have sufficient affinity with monomer or with reaction product.
  • the synthetic reaction of the polypeptide or of the oligonucleotide is proceeded by using the conventional polystyrene-polyethylene glycol grafted support resin for solid- phase reaction
  • the reaction rates and yield are very low compared to those of liquid phase synthesis since the reagent cannot diffuse and penetrate into the bulk of the polymer support.
  • the polystyrene- polyethylene glycol grafted resin in which only the polyethylene glycol phase forms the outer shell layer of the polymeric support, being separated from the polystyrene phase, to be exposed on monomer or solvent, has been developed.
  • linker a part which connects the low molecular weight substrate such as amino acid with the non-soluble polymer support, in the solid-phase synthesis, is called as a linker, in general, which contains a functional group which combines or cleaves the substrate with the polymer support.
  • the linker should be stable in the reaction conditions required for the preparation of the product, and the reaction condition for removal of linker should be mild and the removal reaction should be easily automated.
  • the above linker should maintain the compatibility in all steps for synthesis and should be easily cleavable without any decomposition of reaction product in the recovery condition.
  • the acid-cleavable linkers are the most widely used. However, this type of linker cannot be employed in the acidic reaction condition. In order to obviate this problem, the base-cleavable linker had been developed. However this type of linker also has a problem that it cannot be employed in the reaction of the basic condition.
  • the photo-cleavable linker can be employed in both acidic and basic reaction condition, and thus, can be widely used for various reaction conditions since it can be cleaved by means of light, and has a big merit that the reaction product can be used without any further treatment.
  • a novel technology for the preparation of a polymer support wherein a polyethylene glycol shell which has a good compatibility with the reaction products, and monomers of the polypeptide or oligonucleotide synthesis reaction, is formed on the outer surface of the core supporter to maximize the contact between the reactants and thereby to improve the reaction rate and yield, and moreover, acid-cleavable linker or light-cleavable linker is introduced at the outer end of said shell layer to expose the reaction site on the outer surface to the highest degree in order to improve the reaction rate and yield all the more, has been expected in this technical field.
  • the primary object of the present invention is to provide a process for preparing a gel type core-shell resin, which comprises, a step for swelling the core resin composed of amino methyl polystyrene cross-linked with divinyl benzene, by using a solvent; a step for cross-linking to protect the said amino methyl ends of the amino methyl polystyrene core by reacting triazine with amino methyl polystyrene resin swelled in the above; a step for forming shell layer by reacting the amino methyl polystyrene resin of which amino methyl ends are protected by cross- linking with triazine, with polyalkylene glycol of which both ends are terminated with hydroxy group or amino group.
  • the another purpose of the present invention is to provide a gel type core- shell resin composed of a core section in which amino methyl ends of the amino methy! polystyrene resin cross-linked with divinyl benzene, is cross-linked with cyanuric chloride; and a shell layer made up of polyaikylene glycol resin of which both ends are terminated with hydroxy group or amino group, and combined at the remaining another chloride end of the triazine which is participating with the crosslink structure between the above amino methyl ends.
  • the above primary object of the present invention may be achieved by providing a process for the preparation of a gel type core-shell resin, which comprises i) a step for swelling the core resin composed of amino methyl polystyrene cross-linked with divinylbenzene, by using a solvent; ii) a step for cross- linking to protect the said amino methyl ends by reacting triazi ⁇ e with amino methyl polystyrene resin swelled in the above step i); and iii) a step for forming shell layer by reacting the amino methyl polystyrene resin of which amino methyl ends are protected by cross-linking with triazine, with polyalkylene glycol of which both ends are terminated with hydroxy group or amino group.
  • the preparation method of the core-shell resin of the present invention may further comprises an additional step for combining a photo-cleavable. linker with hydroxy group or amino group which is exposed on the end of the shell surface after the completion of the above step ii).
  • the another purpose of the present invention may be achieved by providing a gel type core-shell resin composed of a core section in which amino methyl ends of the amino methyl polystyrene resin cross-linked with divi ⁇ yl benzene, are cross- linked with cyanuric chloride; and a shell section made up of polyalkylene glycol resin of which both ends are terminated with hydroxy group or amino group and combined at the remaining another chloride end of the triazine which is participating in the crossiink structure between the above amino methyl ends.
  • the amino methyl polystyrene resin the raw material of the core of present invention, had been commercialized long ago, and prepared through the step for forming hydroxy moiety on polystyrene by Friedel Craft alkylation and the step for converting the hydroxy group into amino group by Mitsunobu-Staudi ⁇ ger reaction.
  • the amino methyl polystyrene re ⁇ in of the present invention is swelled with chloroform and then cross-linked with cyanuric chloride, which ⁇ a kind of triazine.
  • Cyanuric chloride is a triazine which has chlorids at 2, 4 and 5 site, and two of chlorids ends thereof react with the amino group of the amino methyl polystyrene.
  • the free amino group does not remain on the surface of the amino methyl polystyrene resin in case of crosslinking the above amino methyl polystyrene resin by using cyanuric chloride. It may be confirmed by Kaiser's test after drying the resin cross-linked with the above cyanuric chloride.
  • the shell layer is formed by combining polyethyieneglycol of which ends are terminated with hydroxy or amino group, with free halide end of the cyanuric halide which js not participating in crosslinking.
  • the polyethylene glycol resin is grafted on the polystyrene core resin of which amine end is protected by cross-linking, the polyethylene glycol resin cannot diffuse into the bulk of amino methyl polystyrene resin cross-linked with divinly benzene due to the immisciblity, the difference in molecular weight and to lack of compatibility between the two resins.
  • the grafting reaction between amino methyl polystyrene core resin and polyethylene glycol resin cannot occur in the bulk phase of the amino methyl polystyrene core resin, and can be proceeded only on the surface of the amino methyl polystyrene core resin, and thereby polyethylene glycol shell layer is formed only on the surface of the above core resin.
  • polystyrene resin and polyethylene glycol resin are blended to make homogeneous phase without phase separation.
  • the polymer support of the present invention is characterized in that polystyrene core resin layer and polyethyieneglycol shell layer are separated with each other.
  • the another purpose of the present invention may be achieved by providing a gel type core-shell resin, which comprises, i) a core resin section composed of amino methyl polystyrene cross-linked with divinyl benzene of which amino methyl ends of the amino methyl polystyrene resin, are cross-linked with cyanuric chloride; and ii) a shell section made up of polyalkylene glycol resin of which both ends are terminated with hydroxy group or amino group and combined with the remaining another chloride end of the triazine which is participating in the crosslink between the above amino methyl ends.
  • a novel core shell type resin which can improve the reaction efficiency and yield in the solid phase synthesis thanks to the functional group which exist only on the outer surface of polymer support, can be prepared.
  • the reaction time can be shortened in case that a compound such as amino acid is introduced on the polymer support of the present invention since the functional group exists only on the surface of the support resin.
  • the polymer support of the present invention can be employed for the various reaction conditions since it is stable in various acidic or basic conditions.
  • the polymer support of the present invention which contains a photo- cleavable linker on the surface of the support resin can be employed for the reaction without any decline of reaction yield due to the light impermeability.
  • Fig. 1 is a reaction scheme showing the process for grafting photo-cleavable linker on the surface of core-shell resin .
  • Fig. 2 is a confocal fluorescent picture of (1 )amino methyl polystyrene resin, (2)TentaGel, and (3)core-shell resin.
  • Fig, 3 is a comparative picture non-specific adsorption of (1)core-shell resin and (2)TentaGel.
  • Fig. 4 is a comparative graph of the time laps required for introduction of the first amino acid between the conventional resin and the core-shell resin of the present invention.
  • Fig. 5 is a comparative graph of photo-cleavable rates between the conventional resin and the core-shell resin of the present invention.
  • the phase of polyethylene glycol resin on which polypeptide or oligonucleotide synthesis reaction are proceeded is separated with that of polystyrene resin. Accordingly, the diffusion and permeation of the monomers and solvent in the shell layer of polyethyle ⁇ eglycol, is easy, and the reaction product can easily be separated from the shell layer.
  • the monomers such as amino acid cannot easily permeat and diffuse into the bulk phase of the polyethylene glycol grafted polystyrene resin of the conventional technology wherein the hydrophobic polystyrene resin is blended with polyethyleneglycol resin, and reaction product formed in the bulk phase of the polymer support, cannot easily be recovered from the support resin prepared by the process of the conventional technology.
  • Fig.2 is a photograph of an amino methyl polystyrene resin
  • (2) of Fig.2 is a photograph of TentaGel
  • (3) of Fig.2 is a picture of core-shell resin which contains amino group formed only on the surface of the support resin.
  • a photo-cleavable linker can be introduced at the hydroxy group or amino group exposed on the end of surface of core-shell resin.
  • the photo-cleavable linkers which can be used for the present invention are the linkers described in Synthetic Peptide-A User's Guide, 2nd Ed., G.A, Grant(Ed), Oxford, pp123-127, 3-nitro-4-hydroxymethylbenzoic acid, 3-nitro-4- amino methyl benzoic acid, 4-(1-ami ⁇ oethyl)-2-methoxy-5-nitro-phenoxypropionic acid and so forth. 4-(1-ami ⁇ oethyl)"2-me1hoxy-5-nitro-pheno ⁇ ybutanoic acid is the most desirable photo-cleavable linker.
  • the photo-cleavable linker of the present invention can be applied on all of the reactions of which condition is acidic or basic, and the reaction product prepared by using the resin of the present invention, can be used without any further treatment.
  • the acid-cleavable linker which can be used in the present invention, and connect the synthesized polypeptide or oligonucleotide with the end of the outer surface of shell layer of the present gel type core shell resin, and can be cleaved easily by the action of an acid to separate the reaction product from the above support resin.
  • the acid-cleavable linker which can be used in the present invention is "Rink amide” linker, "Wang” linker, "SASRiN” linker that already had been known to the pubiicfSynthetic Peptide-A User's Guide", 2nd Ed., G. A. Grant(Ed), Oxford, pp123 ⁇ 127) and so on.
  • amino methyl polystyrene resin cross-linked with diviny! benzene was cross-linked with 2,4,6-trichloro-1 ,3,5-4riazine, cyanuric chloride, after swelling in chloroform(200ml).
  • the amino methyl polystyrene resin(10g, 2.1mmol/g), 2,4,6-trichloro-i ,3 a 5- triazine(19.4g, 105mmol), DIEA(95ml, 105mmol) were placed in a round-bottomed flask, after adding purified chloroform(300ml) as a solvent thereto, and stirred for 12 hours at 50 0 C . After the removal through filtering the solvent after finishing crosslink reaction, cross-linked resin was washed with chloroform(200ml), methanol(200ml) three times of each one and dried for 12hours in vacuum oven. It was confirmed that amino group does not remained in cross-linked resin by using kaiser's test after drying.
  • Fmoc-aminoethyl photo cleavable . linker, 4-(4-(1-Fmoc-amino)ethyl)"2- methoxy ⁇ 5-nitrophe ⁇ oxy)butanoic acid, that amino group is protected with Fmoc, is grafted by using BOB(benzot ⁇ azole-1-yloxy-tris(dimethylamino) ⁇ phosphoniurn hexafluorophosphate)coupli ⁇ g reagent on the core-shell resin in NMP(N- methylpyrrolidone).
  • the core-shell resin(3O0mg 7 0,35 mmol/g) were swollen in NMP(2ml) for 30min, and then photo-cleavable linker(109 mg, 0.21 mmol), BOP(93 mg, 0,21 mmol), HOBt (N-hydroxybenzotriazole, 28mg, 0.21 mmol), DIEA(diisopropyiethylamine, 55/4, 0.32 mmol) were added in NMP(I mI), and then the reaction was performed for 1 hour.
  • Leuenkepali ⁇ amide (YGGFL-N H2) and Fmoc amino acid were synthesized for solid-phase peptide synthesis using BOB coupling reagent in NMP on the core- shell resin grafted photo-cleavable linker prepared in Example 2.
  • the core-shell resin(300 mg, 0.35mmol/g) grafted photo-cleavable linker were swollen for 30 min in NMP(2ml), and then Fmoc amino acid(0.21 mmol), BOP(93mg, 0.21 mmol), HOBt(28 mg, 0.21 mmol), DIEA(55M, 0-32 mmol) were added on the resin in NMP(I mI), and then the reaction was performed for 1 hour,
  • Fmoc amino acid was grafted repeatedly in the order, after the removal Fmoc protecting group of the resin which contains Fmoc amino acid. At this time, 25% piperidi ⁇ e/NMP was treated twice 3min, 7min respectively as a condition of detaching Fmoc.
  • Rink amide linker (p-(R.S)-a-(1 ⁇ (9H-fluoren-9-yl)-methoxyformamido) ⁇ 2,4- dimethoxybenzyl)-phenoxyacetic acid) of which amino group is protected with Fmoc, was grafted by using BOP coupling reagent in NMP(N-methylpyrrolidone) on the core-shell resin.
  • the core shell(300 mg, 0.35mmol/g) resin were swollen in NMP(2ml) for 30 min, and then Rink amide !inker(113 mg, 0.21 mmol), BOP(93 mg, 0.21 mmol), HOBt(N-hydroxybenzotriazole, 28 mg, 0.21 mmol), DIEA(diisopropy ⁇ ethylamine, 55/4, 0.32 mmol) were added in NMP(I mI), and then the reaction was performed for 1 hour. After completion of the reaction, the resin was filtered and washed with DCM(2ml), MeOH(2ml), NMP(2ml) and MeOH(2ml) in the order and dried for 12 hours in vacuum oven.
  • Fmoc amino acid was grafted repeatedly in the order after the removal Fmoc protecting group of the resin which contains Fmoc amino acid.
  • 25% piperidine/NMP was treated twice 3min ( 7min respectively as a condition of detaching Fmoc
  • the resin was treated with 95% TFA, 2.5% H2O, 2.5% TES(triethylsilane) and filtered synthesized peptide.
  • the overall yield was 91 % obtaining peptide and HPLC(high performance liquid chromatography)purity was 98.4%.
  • the core-shell resin(300 mg, 0.35 mmol/g) were swollen for 30 mi ⁇ in NMP(2ml), and then 2-chloro trityl linker(84 mg, 0.21 mmol), BOP(93 mg, 0.21 mmol), HOBt (N-hydroxybenzotriazo)e, 28 mg, 0.21 mmol), DIEA(diisopropyiethylamine ) 55 ⁇ , 0.32 mmol) were added in NMP(I mI) 1 and then the reaction was performed for 1 hour.
  • the resin was filtered and washed with DCM(2ml), MeOH(2ml), NMP(2ml) and MeOH(2tnl) in the order and dried for 12 hours in vacuum oven.
  • the dried resin(315 mg) were swollen in DCM(SmI) repurified for 30 mi ⁇ , and then thionyl chloride(0.5 ml) was added and the reaction was performed for 1.5 hours so that hydroxy group of trityl linker substrated for chloride.
  • the resin was filtered and washed with DCM(2ml) five times and dried for 12 hours in vacuum oven.
  • Leuenkepa ⁇ amide (YGGFL-NH2) was synthesized on the core-shell resin which contains 2-chloro trityl linker prepared in Example 6.
  • the core-shell resin (300mg, 0.35mmol/g) which contains photo-cleavable linker, was swollen for 30 min in NMP(2m(), and then Fmoc amino ac ⁇ d(0.21 mmol), D ⁇ EA(55 ⁇ i, 0.32 mmol) were added in NMP(ImI), and then the reaction was performed for 2 hours.
  • Fmoc amino acid was grafted by using BOP coupling reagent the following removal Fmoc protecting group of the resin which contains Fmoc amino acid,
  • the above re ⁇ in(300mg, 0.35mmol/g) which contains amino group, were swollen for 30mi ⁇ in NMP(2mI), Fmoc amino acid(0.21 mmol), BOp(93 mg, 0.21 mmol), HOBt(28 mg, 0.21 mmol), DIEA(55 / 4, 0.32 mmol) were added in NMP(I mI), the reaction was performed for 1 hour, Fmoc amino acid was grafted repeatedly in the order after the removal Fmoc protecting group of the resin which contained Fmoc amino acid.
  • FITC(5-fluorescein isothiocyanate),a fluorescent material were grafted on the amino methyl polystyrene resin which already has been used commercialized, TentaGel and the core-shell resin, and taken the co ⁇ focal fluorescent photograph to demonstrate that the core-shell resin is the core-shell resin. 10rng of each resin was placed in 1 ,5ml Eppendorf tube and swollen for 30min in NMP(0.5ml). FITC( 8 mg, 20 //mo!), DIEA (3.5 ⁇ JL ⁇ 20 /zmol) were added in NMP(5ml), and then the reaction was performed for 12 hours at room temperature.
  • Fig. 2 The confocal fluorescent photograph is described in Fig. 2.
  • (1 ) of Fig. 2 is a photo of amino methyl polystyrene resin
  • (2) of Fig. 2 is a photo of TentaGel
  • (3) of Fig. 2 is a photo of core-shell resin.
  • the core-shell resin was confirmed by Fig. 3.
  • Non-specific binding is a significant limitation to biochemical assay such as protein or DNA analysis of resin. For this reason, many studies about resolving this problem have been performed.
  • Non-specific binding of core-shell resin was compared with TentaGel comprising PEG that reduce the non-specific binding effectively.
  • TentaGel and core-shell resin 5mg of each one were placed in vial, and then acetic anhydride 30 equiv. and DiEA were added in NMP(I mI), and then the reaction was performed for 2 hours and all amino group of each resin were acetylated.
  • the beads 1mg of each one such that amino group, was acetylated were placed in Eppendorf tube, and then 1ml Tris-buffer(100 mM, pH 7.5) was added and made suspension, and then 1 ⁇ (alkaline phosphatase 1 units) ST- AP(Streptavidin-alkaline phosphatase) solution was added and the reaction was performed for 30 min at 30"C . After filtering the following the reaction, the bead was washed with tris-buffer(100 mM, pH 9.5) five times and suspended in 1ml Tris- buffer(100 mM, pH 9.5).
  • Fig. 3 is a photograph for comparing non-specific binding and the picture(1 )(2) were performed with core-shell resin, TentaGel respectively. As seen in the above Fig.3, core-shell resin have non-specific binding on their surface, it was shown the possibility of using such as protein or DNA analysis.
  • Fig. 4 is the comparative graph of loading time of the first amino acid on core-shell resin, TentaGel, amino methyl polystyrene resin, and we found that the loading ⁇ f the first amino acid on the core-shell resin can be proceeded much more efficiently than that on the other resins.
  • Photo-cleavable linker was grafted on the amino methyl polystyrene resin which has been commercialized, TentaGel same as core-shell resin of Example 2 and Leu-enkepalinamide was grafted same as Example 4. 100mg of each resin were placed in vial(5ml), and then methano
  • the comparative graph of photo-cleavable rates among core-shell resin, TentaGel, amino methyl polystyrene resin, amino methyl polystyrene resin in the middle of the conventional resin released less than 40% of the product even after 3 hours, in case of TentaGel that is well swollen in methanol due to the loading of PEG released 70% of the product after 2 hours and the yield was no longer increased,
  • the core-shell resin released the product rapidly and gave more than 95% yield within 1 hour unlike the other gel-type resins.
  • the present invention provide a process for the preparation of the gel type core-shell resin, which comprises, a step for swelling core
  • a novel core shell type resin which can improve the reaction rate and yield in the solid phase synthesis for various kind of organic compound since the function group exists only on the surface of the support polymer.
  • the core shell resin of the present invention can be applied on various kinds of organic reaction regardless the acidity condition, and can shorten the time for the introduction of amino acid on the polymer support since the function group exist only on the surface of the polymer support.

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  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
  • Other Resins Obtained By Reactions Not Involving Carbon-To-Carbon Unsaturated Bonds (AREA)

Abstract

L'invention concerne une résine à structure coeur-écorce ('core-shell') de type gel. Elle concerne en particulier une résine à structure coeur-écorce de type gel, comprenant: une partie coeur dans laquelle les extrémités aminométhyle de la résine d'aminométhylpolystyrène réticulée avec du divinylbenzène sont réticulées avec du chlorure cyanurique; et une partie écorce constituée d'une résine de polylalkylène glycol à terminaison hydroxy ou amino aux deux extrémités et combinée à l'extrémité chlorure restante de la triazine participant à la liaison réticulaire entre les extrémités aminométhyle mentionnées ci-dessus.
PCT/KR2005/003273 2004-10-04 2005-10-04 Resine a structure coeur-ecorce ('core-shell') de type gel WO2006126760A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020040078860A KR101088579B1 (ko) 2004-10-04 2004-10-04 젤 타입의 코어-쉘 수지
KR10-2004-0078860 2004-10-04

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102498123A (zh) * 2009-07-15 2012-06-13 新加坡科技研究局 改进的生物聚合物筛选

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