WO2002001223A1 - Substructure approach to molecularly imprinted polymers with high selectivity for folic acid and analogues - Google Patents
Substructure approach to molecularly imprinted polymers with high selectivity for folic acid and analogues Download PDFInfo
- Publication number
- WO2002001223A1 WO2002001223A1 PCT/SE2001/001483 SE0101483W WO0201223A1 WO 2002001223 A1 WO2002001223 A1 WO 2002001223A1 SE 0101483 W SE0101483 W SE 0101483W WO 0201223 A1 WO0201223 A1 WO 0201223A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- analogues
- template
- folic acid
- molecularly imprinted
- approach
- Prior art date
Links
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 title claims abstract description 18
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 title claims abstract description 9
- 229960000304 folic acid Drugs 0.000 title claims abstract description 9
- 235000019152 folic acid Nutrition 0.000 title claims abstract description 9
- 239000011724 folic acid Substances 0.000 title claims abstract description 9
- 229920000344 molecularly imprinted polymer Polymers 0.000 title abstract description 9
- 238000013459 approach Methods 0.000 title description 6
- 239000000178 monomer Substances 0.000 claims abstract description 23
- 229920000642 polymer Polymers 0.000 claims description 18
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 3
- 235000013922 glutamic acid Nutrition 0.000 claims description 3
- 239000004220 glutamic acid Substances 0.000 claims description 3
- 150000003195 pteridines Chemical class 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 description 13
- 238000000926 separation method Methods 0.000 description 10
- 238000000034 method Methods 0.000 description 7
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 6
- 230000027455 binding Effects 0.000 description 6
- 238000004132 cross linking Methods 0.000 description 6
- DBCAQXHNJOFNGC-UHFFFAOYSA-N 4-bromo-1,1,1-trifluorobutane Chemical compound FC(F)(F)CCCBr DBCAQXHNJOFNGC-UHFFFAOYSA-N 0.000 description 5
- STVZJERGLQHEKB-UHFFFAOYSA-N ethylene glycol dimethacrylate Substances CC(=C)C(=O)OCCOC(=O)C(C)=C STVZJERGLQHEKB-UHFFFAOYSA-N 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000010526 radical polymerization reaction Methods 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- OZAIFHULBGXAKX-UHFFFAOYSA-N 2-(2-cyanopropan-2-yldiazenyl)-2-methylpropanenitrile Chemical compound N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 description 4
- 238000006116 polymerization reaction Methods 0.000 description 4
- NOYPYLRCIDNJJB-UHFFFAOYSA-N trimetrexate Chemical compound COC1=C(OC)C(OC)=CC(NCC=2C(=C3C(N)=NC(N)=NC3=CC=2)C)=C1 NOYPYLRCIDNJJB-UHFFFAOYSA-N 0.000 description 4
- 229960001099 trimetrexate Drugs 0.000 description 4
- DSPLSFBRERHHIN-AWEZNQCLSA-N (2s)-2-amino-n,3-diphenylpropanamide Chemical compound C([C@H](N)C(=O)NC=1C=CC=CC=1)C1=CC=CC=C1 DSPLSFBRERHHIN-AWEZNQCLSA-N 0.000 description 3
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 3
- 230000005526 G1 to G0 transition Effects 0.000 description 3
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000003999 initiator Substances 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 125000005395 methacrylic acid group Chemical group 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- LIVXWXAMTVJGCO-UHFFFAOYSA-N 2,4-diamino-6,7-diisopropylpteridine Chemical compound NC1=NC(N)=C2N=C(C(C)C)C(C(C)C)=NC2=N1 LIVXWXAMTVJGCO-UHFFFAOYSA-N 0.000 description 2
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 2
- ABLZXFCXXLZCGV-UHFFFAOYSA-N Phosphorous acid Chemical class OP(O)=O ABLZXFCXXLZCGV-UHFFFAOYSA-N 0.000 description 2
- OKKRPWIIYQTPQF-UHFFFAOYSA-N Trimethylolpropane trimethacrylate Chemical compound CC(=C)C(=O)OCC(CC)(COC(=O)C(C)=C)COC(=O)C(C)=C OKKRPWIIYQTPQF-UHFFFAOYSA-N 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 238000000429 assembly Methods 0.000 description 2
- 230000000712 assembly Effects 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 229960000485 methotrexate Drugs 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 125000002348 vinylic group Chemical group 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- PVFCXMDXBIEMQG-JTQLQIEISA-N (2s)-2-(phenylmethoxycarbonylamino)pentanedioic acid Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)OCC1=CC=CC=C1 PVFCXMDXBIEMQG-JTQLQIEISA-N 0.000 description 1
- 125000000143 2-carboxyethyl group Chemical group [H]OC(=O)C([H])([H])C([H])([H])* 0.000 description 1
- PNKUSGQVOMIXLU-UHFFFAOYSA-N Formamidine Chemical compound NC=N PNKUSGQVOMIXLU-UHFFFAOYSA-N 0.000 description 1
- 206010022528 Interactions Diseases 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000003862 amino acid derivatives Chemical class 0.000 description 1
- -1 amino acid enantiomers Chemical class 0.000 description 1
- 150000001414 amino alcohols Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 238000002045 capillary electrochromatography Methods 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000006897 homolysis reaction Methods 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229940116254 phosphonic acid Drugs 0.000 description 1
- 229960004838 phosphoric acid Drugs 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- CPNGPNLZQNNVQM-UHFFFAOYSA-N pteridine Chemical group N1=CN=CC2=NC=CN=C21 CPNGPNLZQNNVQM-UHFFFAOYSA-N 0.000 description 1
- CITCTUNIFJOTHI-UHFFFAOYSA-N pteridine-2,4-diamine Chemical class N1=CC=NC2=NC(N)=NC(N)=C21 CITCTUNIFJOTHI-UHFFFAOYSA-N 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 229920001187 thermosetting polymer Polymers 0.000 description 1
- IEDVJHCEMCRBQM-UHFFFAOYSA-N trimethoprim Chemical compound COC1=C(OC)C(OC)=CC(CC=2C(=NC(N)=NC=2)N)=C1 IEDVJHCEMCRBQM-UHFFFAOYSA-N 0.000 description 1
- 229960001082 trimethoprim Drugs 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/82—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving vitamins or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2600/00—Assays involving molecular imprinted polymers/polymers created around a molecular template
Definitions
- MIPs Molecularly imprinted polymers
- the technique shows promise in chiral separations of for example amino acid derivatives, peptides, phosphonates, aminoalcohols and beta-blocking compounds, affinity chromatography of nucleotides and the DNA-bases as well as substitute for antibodies in immunoassays or extractions of commercial drugs.
- MI Molecular imprinting
- the separation can be through an affinity chromatographic procedure where pH, ion strength or solvent gradients can be used in order to control the strength of interaction with the stationary phase.
- the separation can target enantiomers or diastereomers in a mixture of enantiomers or diastereomers of one or many compounds.
- Analytical applications can in addition to the above mentioned separations be: competetitive binding assays, chemical sensors or selective sample enrichments . 1
- the materials can be synthesized in any standard equipped laboratory in a relatively short time and some of the MIPs exhibit binding affinities and selectivities in the order of those exhibited by antibodies towards their antigens.
- Most MIPs are synthesized by free radical polymerization of functional monounsaturated (vinylic, acrylic, methacrylic) monomers and an excess of crosslinking di- or tri- unsaturated (vinylic, acrylic, methacrylic) monomers resulting in porous organic network materials.
- These polymerizations have the advantage of being relatively robust allowing polymers to be prepared in high yield using different solvents (aqueous or organic) and at different temperatures. This is necessary in view of the varying solubilities of the template molecules.
- the most successful noncovalent imprinting systems are based on commodity acrylic or methacrylic monomers, such as methacrylic acid (MAA) , crosslinked with ethyleneglycol dimethacrylate (EDMA) .
- MAA methacrylic acid
- EDMA ethyleneglycol dimethacrylate
- L-PA L-phenylalanine anilide
- the template (L-PA) , the functional monomer (MAA) and the crosslinking monomer (EDMA) are dissolved in a poorly hydrogen bonding solvent (diluent) of low to medium polarity.
- the free radical polymerization is then initiated with an azo initiator, commonly azo-N,N' -bis-isobutyronitrile (AIBN) either by photochemical homolysis below room temperature or thermochemically at 60°C or higher.
- AIBN azo-N,N' -bis-isobutyronitrile
- the resultant polymer is crushed by mortar and pestle or in a ball mill, extracted by a Soxhlet apparatus, and sieved to a particle size suitable for chromatographic
- the polymers are then evaluated as stationary phases in chromatography by comparing the retention time or capacity factor (k' ) of the template with that of structurally related analogs.
- This invention describes the synthesis of a new class of molecularly imprinted polymers capable of recognizing folic acid and its analogues ( Figure 2) .
- Figure 2 For this purpose specially designed templates and functional monomers have been used.
- the poor solubility and stability of these templates require alternative approaches to be developed.
- substructures of these compounds may be targeted using organic soluble analogues or newly designed monomers .
- Particularly strong binding was obtained when using methacrylic acid as the functional monomer and organic soluble diaminopteridine analogues as templates ( Figure 3) , thus targeting the pteridine ring system.
- analogues can be 2 , 4-diamino-6, 7-diisopropylpteri- dine (DIP) , trimethopriim (TRP) or trimetrexate (TRX) .
- DIP diisopropylpteri- dine
- TRP trimethopriim
- TRX trimetrexate
- the polymer is synthesized by free radical polymerization of a mixture of methacrylic acid or other functional monomer, and a crosslinking monomer, that can be ethyleneglycoldimethacrylate or trimethylolpropane- trimethacrylate, in presence of a solvent and a template and an initiator, that can be azobisisobutyronitrile.
- the template can be 2 , 4-diamino-6, 7-diisopropylpteridine (DIP) , trimethoprim (TRP) or trimetrexate (TRX) . ( Figure 3) After polymerization the polymer is freed from the template by a washing procedure and can then be used for selective separations.
- the polymer is synthesized by free radical polymerization of a mixture of a formamidine or any of the functional monomers in Figure 4, targeting the glutamic acid side chain, and a crosslinking monomer, that can be ethyleneglycoldimethacrylate or trimethylolpropanetrimeth- acrylate, in presence of a solvent and a template and an initiator, that can be azobisisobutyronitrile.
- the template can be Glutamic acid or an analogue thereof, folic acid, methotrexate or leucovorine or analogues of these. After polymerization the polymer is freed from the template by a washing procedure and can then be used for selective separations.
- the polymer prepared according to Example 1 and 2 can be used for separation of enantiomers or diastereomers of the template or for separation of the template or template analogues from structurally related compounds. This can be done by chromatography, capillary electrophoresis, capillary electrochromatography, batch modes or membrane modes .
- the polymer can further be used for catalysing chemical reactions such as esterolysis, amidolysis, ester synthesis or amide synthesis or used in chemical sensors . Litterature
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Peptides Or Proteins (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Steroid Compounds (AREA)
Abstract
The invention refers to templates and functional monomers that can be used to generate molecularly imprinted polymers for the selective recognition of folic acid and its analogues.
Description
SUBSTRUCTURE APPROACH TO MOLECULARLY IMPRINTED POLYMERS WITH HIGH SELECTIVITY FOR FOLIC ACID AND ANALOGUES
In the fields of medical-, dietary-, environmental- and chemical sciences there is an increasing need for the selective separations of specific substances in complex mixtures of related substances. The end goal can be the preparative isolation of a certain compound or compounds, their selective removal from various environments or measurements of their concentration. Molecularly imprinted polymers (MIPs) exhibit often a high selectivity towards their substrate in analogy the antibody-antigen complementarit . ^ The technique shows promise in chiral separations of for example amino acid derivatives, peptides, phosphonates, aminoalcohols and beta-blocking compounds, affinity chromatography of nucleotides and the DNA-bases as well as substitute for antibodies in immunoassays or extractions of commercial drugs.2 Molecular imprinting (MI) consists of the following key steps (Figure 1) : (1) Functional monomers are allowed to interact reversibly with a template molecule in solution. (2) The hereby formed template assemblies are compolymerized with a crosslinking monomer resulting in a crosslinked network polymer. (3) The template is displaced and the materials can be used for selective molecular recognition of the corresponding compound. If these are crushed and sieved they can be packed in a chromatographic column and used for chromatographic separation of the template from structurally related analogs. Analytical as well as preparative applications are here possible. Preparative applications can be separation of a compound from a complex mixture of structurally related compounds and isolation of the compound. This can be through an affinity chromatographic procedure where pH, ion strength or solvent gradients can be used in order to control the
strength of interaction with the stationary phase. The separation can target enantiomers or diastereomers in a mixture of enantiomers or diastereomers of one or many compounds. Analytical applications can in addition to the above mentioned separations be: competetitive binding assays, chemical sensors or selective sample enrichments .1
Currently the most widely applied technique to generate molecularly imprinted binding sites is represented by the noncovalent route.3 This makes use of noncovalent self-assembly of the template with functional monomers prior to polymerization, free radical polymerization with a crosslinking monomer and then template extraction followed by rebinding by noncovalent inter- actions. Although the preparation of a MIP by this method is technically simple it relies on the success of stabilisation of the relatively weak interactions between the template and the functional monomers. Stable monomer-template assemblies will in turn lead to a larger concentration of high affinity binding sites in the resulting polymer. The materials can be synthesized in any standard equipped laboratory in a relatively short time and some of the MIPs exhibit binding affinities and selectivities in the order of those exhibited by antibodies towards their antigens. Most MIPs are synthesized by free radical polymerization of functional monounsaturated (vinylic, acrylic, methacrylic) monomers and an excess of crosslinking di- or tri- unsaturated (vinylic, acrylic, methacrylic) monomers resulting in porous organic network materials. These polymerizations have the advantage of being relatively robust allowing polymers to be prepared in high yield using different solvents (aqueous or organic) and at different temperatures. This is necessary in view of the varying solubilities of the template molecules.
The most successful noncovalent imprinting systems are based on commodity acrylic or methacrylic monomers,
such as methacrylic acid (MAA) , crosslinked with ethyleneglycol dimethacrylate (EDMA) . Initially, derivatives of amino acid enantiomers were used as templates for the preparation of imprinted stationary phases for chiral separations (MICSPs) but this system has proven generally applicable to the imprinting of templates allowing hydrogen bonding or electrostatic interactions to develop with MAA.4 The procedure has been applied to the imprinting with L-phenylalanine anilide (L-PA) . In the first step, the template (L-PA) , the functional monomer (MAA) and the crosslinking monomer (EDMA) are dissolved in a poorly hydrogen bonding solvent (diluent) of low to medium polarity. The free radical polymerization is then initiated with an azo initiator, commonly azo-N,N' -bis-isobutyronitrile (AIBN) either by photochemical homolysis below room temperature or thermochemically at 60°C or higher. In the final step, the resultant polymer is crushed by mortar and pestle or in a ball mill, extracted by a Soxhlet apparatus, and sieved to a particle size suitable for chromatographic
(25-38 μm) or batch (150-250 μm) applications. The polymers are then evaluated as stationary phases in chromatography by comparing the retention time or capacity factor (k' ) of the template with that of structurally related analogs.
In spite of the fact that the MIPs can recognize a large number of structures used as templates, a number of compounds classes are only poorly recognized by polymers prepared using the present imprinting protocols . This is the case of several biomolecules i.e. vitamins, antibiotics, cofactors, proteins, peptides, several of which is soluble only in aqueous solvents, not suited for molecular imprinting with the above protocol . Furthermore the binding strength between the functional monomer and the template is often insufficient leading to a low sample load capacity and a significant nonspecific binding. There is therefore a need for the development
of new approaches including the use of specially designed template analogues and functional monomers binding stronger to the template and allowing recognition of new compound classes . For instance monomers designed to bind carboxylic-, phosphoric- or phosphonic- acid templates are needed.
This invention describes the synthesis of a new class of molecularly imprinted polymers capable of recognizing folic acid and its analogues (Figure 2) . For this purpose specially designed templates and functional monomers have been used. In order to synthesise polymers with a high selectivity and affinity for folic acid and its analogues, the poor solubility and stability of these templates require alternative approaches to be developed. Thus substructures of these compounds may be targeted using organic soluble analogues or newly designed monomers . Particularly strong binding was obtained when using methacrylic acid as the functional monomer and organic soluble diaminopteridine analogues as templates (Figure 3) , thus targeting the pteridine ring system.
These analogues can be 2 , 4-diamino-6, 7-diisopropylpteri- dine (DIP) , trimethopriim (TRP) or trimetrexate (TRX) . In a separate approach targeting the glutamic acid substructure of folic acid and analogues thereof, N-Cbz- glutamic acid was used as template in combination with a new class of amidinebased functional monomers (Figure 4) . These approaches resulted in polymers showing high selectivity for the target compounds. The polymers were evaluated by HPLC by injecting the template methotrexate, leucovorine and folic acid. After a detailed mobile phase optimization, systems were found allowing strong and highly selective retentions in aqueous mobile phases .
The invention will now be described in more detail giving a number of nonrestricting examples:
Example 1
The polymer is synthesized by free radical polymerization of a mixture of methacrylic acid or other functional monomer, and a crosslinking monomer, that can be ethyleneglycoldimethacrylate or trimethylolpropane- trimethacrylate, in presence of a solvent and a template and an initiator, that can be azobisisobutyronitrile. The template can be 2 , 4-diamino-6, 7-diisopropylpteridine (DIP) , trimethoprim (TRP) or trimetrexate (TRX) . (Figure 3) After polymerization the polymer is freed from the template by a washing procedure and can then be used for selective separations.
Example 2
The polymer is synthesized by free radical polymerization of a mixture of a formamidine or any of the functional monomers in Figure 4, targeting the glutamic acid side chain, and a crosslinking monomer, that can be ethyleneglycoldimethacrylate or trimethylolpropanetrimeth- acrylate, in presence of a solvent and a template and an initiator, that can be azobisisobutyronitrile. The template can be Glutamic acid or an analogue thereof, folic acid, methotrexate or leucovorine or analogues of these. After polymerization the polymer is freed from the template by a washing procedure and can then be used for selective separations. Example 3
The polymer prepared according to Example 1 and 2 can be used for separation of enantiomers or diastereomers of the template or for separation of the template or template analogues from structurally related compounds. This can be done by chromatography, capillary electrophoresis, capillary electrochromatography, batch modes or membrane modes . The polymer can further be used for catalysing chemical reactions such as esterolysis, amidolysis, ester synthesis or amide synthesis or used in chemical sensors .
Litterature
1. Bartsch, R.A. & Maeda, M. in ACS Symposium Series 703 (Oxford University Press, Washington, 1998) .
2. Andersson, L.I., Mύller, R. , Vlatakis, G. & Mosbach, K. Proc . Natl . Acad . Sci . U. S . A . 92, 4788-92 (1995) .
3. Sellergren, B., Lepistoe, M. & Mosbach, K. J". Am . Chem. Soc . 110, 5853-60 (1988) .
4. Sellergren, B. in A practical approach to chiral separation by liquid chromatography (ed. Subramanian, G.) 69-93 (VCH, Weinheim, 1994) .
Claims
1. A polymer selective for folic acid or analogues thereof .
2. A polymer according to claim 1 characterised in that it is synthesised in presence of pteridine analogues .
3. A polymer according to claim 1 characterised in that it is synthesised in presence of glutamic acid or derivatives thereof.
4. A polymer according to claim 1 characterised in that it is synthesised in presence of amidinebased functional monomers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2001267970A AU2001267970A1 (en) | 2000-06-28 | 2001-06-28 | Substructure approach to molecularly imprinted polymers with high selectivity for folic acid and analogues |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE0002463-8 | 2000-06-28 | ||
| SE0002463A SE0002463D0 (en) | 2000-06-28 | 2000-06-28 | Substructure approach to molecular imprinted polymers with high selectivity for folic acid and analogues |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2002001223A1 true WO2002001223A1 (en) | 2002-01-03 |
Family
ID=20280313
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/SE2001/001483 WO2002001223A1 (en) | 2000-06-28 | 2001-06-28 | Substructure approach to molecularly imprinted polymers with high selectivity for folic acid and analogues |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU2001267970A1 (en) |
| SE (1) | SE0002463D0 (en) |
| WO (1) | WO2002001223A1 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004067578A1 (en) * | 2003-01-30 | 2004-08-12 | Mip Technologies Ab | Moleculary imprinted polymers for extraction of components from foodstruffs |
| WO2006041398A1 (en) * | 2004-10-12 | 2006-04-20 | Mip Technologies Ab | Method for producing molecularly imprinted polymers for the recognition of target molecules |
| US20110002874A1 (en) * | 2005-04-28 | 2011-01-06 | Board Of Regents, The University Of Texas System | Polymer Network Compositions and Associated Methods |
| CN104267087A (en) * | 2014-10-29 | 2015-01-07 | 安徽师范大学 | Electrochemical biosensor as well as preparation method and application thereof |
| CN105693960A (en) * | 2016-03-03 | 2016-06-22 | 华南师范大学 | Method for preparing glutamic acid surface molecularly imprinted polymer silica microspheres |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999047531A1 (en) * | 1998-03-16 | 1999-09-23 | Chiral Technologies, Inc. | Chiral separations of pyrimidines |
-
2000
- 2000-06-28 SE SE0002463A patent/SE0002463D0/en unknown
-
2001
- 2001-06-28 AU AU2001267970A patent/AU2001267970A1/en not_active Abandoned
- 2001-06-28 WO PCT/SE2001/001483 patent/WO2002001223A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999047531A1 (en) * | 1998-03-16 | 1999-09-23 | Chiral Technologies, Inc. | Chiral separations of pyrimidines |
Non-Patent Citations (7)
| Title |
|---|
| CONG YU ET AL.: "Enantiomeric recognition by molecularly imprinted polymers using hydrophobic interactions", ANALYTICAL LETTERS, vol. 30, no. 12, 1997, pages 2123 - 2140, XP002949451 * |
| CONG YU ET AL.: "Insights into the origins of binding and the recognition properties of molecularly imprinted polymers prepared using an amide as the hydrogen-bonding functional group", JOURNAL OF MOLECULAR RECOGNITION, vol. 11, 1998, pages 69 - 74, XP002905461 * |
| LARS I. ANDERSSON ET AL.: "Enantiomeric resolution on molecularly imprinted polymers prepared with only non-covalent and non-ionic interactions", JOURNAL OF CHROMATOGRAPHY, vol. 516, 1990, pages 313 - 322, XP002949450 * |
| MARIA KEMPE ET AL.: "Chiral separation using molecularly imprinted heteroaromatic polymers", JOURNAL OF MOLECULAR RECOGNITION, vol. 6, 1993, pages 25 - 29, XP002949454 * |
| MASAKAZU YOSHIKAWA ET AL.: "Alternative molecularly imprinted membranes from a derivative of natural polymer, cellulose acetate", JOURNAL OF APPLIED POLYMER SCIENCE, vol. 72, 1999, pages 493 - 499, XP002949452 * |
| MILENA QUAGLIA ET AL.: "Target analogue imprinted polymers with affinity for folic acid and related compounds", J. AM. CHEM. SOC., vol. 123, 2001, pages 2146 - 2154, XP002949449 * |
| SHOUZHUO YAO ET AL.: "Biomimetic bulk acoustic wave sensor for determination of trimethoprim in the organic phase based on a molecular imprinting polymer", ANALYTICAL SCIENCES, vol. 16, February 2000 (2000-02-01), pages 211 - 215, XP002949453 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004067578A1 (en) * | 2003-01-30 | 2004-08-12 | Mip Technologies Ab | Moleculary imprinted polymers for extraction of components from foodstruffs |
| US7750090B2 (en) | 2003-01-30 | 2010-07-06 | Mip Technologies Ab | Moleculary imprinted polymers for extraction of components from foodstruffs |
| WO2006041398A1 (en) * | 2004-10-12 | 2006-04-20 | Mip Technologies Ab | Method for producing molecularly imprinted polymers for the recognition of target molecules |
| US20110002874A1 (en) * | 2005-04-28 | 2011-01-06 | Board Of Regents, The University Of Texas System | Polymer Network Compositions and Associated Methods |
| CN104267087A (en) * | 2014-10-29 | 2015-01-07 | 安徽师范大学 | Electrochemical biosensor as well as preparation method and application thereof |
| CN105693960A (en) * | 2016-03-03 | 2016-06-22 | 华南师范大学 | Method for preparing glutamic acid surface molecularly imprinted polymer silica microspheres |
| CN105693960B (en) * | 2016-03-03 | 2018-07-17 | 华南师范大学 | A kind of preparation method of glutamic acid molecular imprinted polymer on surface silica gel microball |
Also Published As
| Publication number | Publication date |
|---|---|
| AU2001267970A1 (en) | 2002-01-08 |
| SE0002463D0 (en) | 2000-06-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1250315B1 (en) | Functional monomers for molecular recognition and catalysis | |
| Sellergren | Polymer-and template-related factors influencing the efficiency in molecularly imprinted solid-phase extractions | |
| Rachkov et al. | Recognition of oxytocin and oxytocin-related peptides in aqueous media using a molecularly imprinted polymer synthesized by the epitope approach | |
| Yilmaz et al. | Influence of functional and cross-linking monomers and the amount of template on the performance of molecularly imprinted polymers in binding assays | |
| US6582971B1 (en) | Imprinting large molecular weight compounds in polymer composites | |
| Kempe et al. | Chiral separation using molecularly imprinted heteroaromatic polymers | |
| Zheng et al. | Sulfonamide imprinted polymers using co-functional monomers | |
| Rachkov et al. | Molecularly imprinted polymers prepared in aqueous solution selective for [Sar1, Ala8] angiotensin II | |
| WO1998054228A2 (en) | Amide containing molecular imprinted polymers | |
| JP2002511924A (en) | Cation exchange stationary phase based on bifunctional crown ether for liquid chromatography | |
| Yemiş et al. | Molecularly imprinted polymers and their synthesis by different methods | |
| Dmitrienko et al. | Use of molecular imprinted polymers for the separation and preconcentration of organic compounds | |
| CA2513429C (en) | Molecularly imprinted polymers for extraction of components from foodstuffs | |
| Piletsky et al. | Application of non-specific fluorescent dyes for monitoring enantio-selective ligand binding to molecularly imprinted polymers | |
| Yang et al. | An artificial receptor synthesized by surface-confined imprinting for the recognition of acetylation on histone H4 K16 | |
| Ansell et al. | Molecularly imprinted polymer combinatorial libraries for multiple simultaneous chiral separations | |
| WO2002001223A1 (en) | Substructure approach to molecularly imprinted polymers with high selectivity for folic acid and analogues | |
| Chen et al. | The specificity of a chlorphenamine–imprinted polymer and its application | |
| Haginaka et al. | Uniform-sized molecularly imprinted polymer material for propranolol. Recognition of propranolol and its metabolites | |
| Lai et al. | Chromatographic characterization of molecularly imprinted microspheres for the separation and determination of trimethoprim in aqueous buffers | |
| Chen et al. | Molecular recognition of procainamide-imprinted polymer | |
| Nomura et al. | Selective recognition of 2, 4-dichlorophenoxyacetic acid using a molecularly imprinted polymer | |
| US20030219828A1 (en) | Imprinting large molecular weight compounds in polymer composites | |
| Andersson et al. | Crown ethers as a tool for the preparation of molecularly imprinted polymers | |
| Hamase et al. | Enantio-selective derivatization of amino compounds in the presence of a molecular imprint polymer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ CZ DE DE DK DK DM DZ EC EE EE ES FI FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
| DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
| REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
| 122 | Ep: pct application non-entry in european phase | ||
| NENP | Non-entry into the national phase |
Ref country code: JP |