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WO1995001347A1 - Nouveaux agents reticulants et leur utilisation - Google Patents

Nouveaux agents reticulants et leur utilisation Download PDF

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Publication number
WO1995001347A1
WO1995001347A1 PCT/FI1994/000293 FI9400293W WO9501347A1 WO 1995001347 A1 WO1995001347 A1 WO 1995001347A1 FI 9400293 W FI9400293 W FI 9400293W WO 9501347 A1 WO9501347 A1 WO 9501347A1
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WIPO (PCT)
Prior art keywords
preparation
polymer
amino acid
formula
compound
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Application number
PCT/FI1994/000293
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English (en)
Inventor
Pirjo Korhonen
Original Assignee
Cultor Oy
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 Cultor Oy filed Critical Cultor Oy
Priority to AU70756/94A priority Critical patent/AU7075694A/en
Priority to EP94919700A priority patent/EP0706520A1/fr
Publication of WO1995001347A1 publication Critical patent/WO1995001347A1/fr
Priority to FI956318A priority patent/FI956318L/fi

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
    • C07D295/185—Radicals derived from carboxylic acids from aliphatic carboxylic acids
    • 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
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/268—Polymers created by use of a template, e.g. molecularly imprinted polymers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D229/00—Heterocyclic compounds containing rings of less than five members having two nitrogen atoms as the only ring hetero atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/06—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members
    • C07D241/08—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members with oxygen atoms directly attached to ring carbon atoms
    • 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
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
    • C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/12—Cyclic peptides with only normal peptide bonds in the ring
    • 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
    • C08F20/00—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride, ester, amide, imide or nitrile thereof
    • C08F20/02—Monocarboxylic acids having less than ten carbon atoms, Derivatives thereof
    • C08F20/52—Amides or imides
    • C08F20/54—Amides, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide
    • C08F20/60—Amides, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide containing nitrogen in addition to the carbonamido nitrogen
    • 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

  • Water soluble cross-linking agents are of great industrial interest for the preparation of mechanically stable water compatible polymers.
  • a high degree of cross-linking is necessary when polymerizing, for example, in water. This in turn requires a high water solubility of the molecule functioning as cross-linker.
  • hydrophilic polymer matrices e.g. Sephadex and Sepharose (both products from Pharmacia) are well suited for polar water soluble compounds.
  • Another conventional separation technique for water soluble molecu ⁇ les, electrophoresis is common for the separation of peptides and proteins.
  • electrophoresis N,N'-methylene bisacrylamide is often used as a cross-linking agent, in many cases the cross-linking agent 1,4-bisacryloylpipera- zine, which is closely related to this invention, has given better results 3 .
  • hydrophilic mechanically stable polymers has given improved results in the so-called solid phase synthesis of peptides 4 .
  • This invention relates to new cross-linking agents of the formula I
  • rings (a) to (d) can carry 1 to n substituents, whe ⁇ rein n equals the number of protonated carbon atoms in the ring, the substituents being selected from the group consisting of -OH, -SH, -CN, Cj-C 3 -alkyl and hydroxy-C ! -C 3 - alkyl, or in the rings (a) to (c) , one or more of the -du ⁇ ring members can be replaced by a carbonyl group -C(O)-, and R 1 is selected from the group consisting of C ⁇ -C 3 -alkyl or hydroxy.
  • X is unsubstituted piperazine and R 1 is methyl, i.e. l-acryloyl-4-methacryloylpiperazi- ne, or l-[l-oxo-2-methyl-2-propenyl]-4-[l-oxo-2-propenyl]- piperazine, sum formula C ⁇ H 16 N 2 0 2 .
  • l-acryloyl-4-methacryloyl- piperazine has an unusually high water solubility (2 kg/li ⁇ ter at room temperature) and is thus very useful for the preparation of strongly cross-linked water compatible polymers
  • l-acryloyl-4-methacryloylpiperazine has a water solubility which is 667 times higher (per weight) than the often used cross-linking agent N,N'-methylenebisacrylamide (solubility 3.0 g/liter at room temperature 2 .
  • N,N'-methylenebisacrylamide solubility 3.0 g/liter at room temperature 2
  • l-acryloyl-4-methacryloylpiperazine has a high solubility in organic solvents such as e.g. chloro- form (2 kg/liter at room temperature) , wherefore this cross-linking agent also can be used for the preparation of polymers in organic solvents.
  • the invention also relates to the preparation of the novel cross-linking agents and their use for the preparation of polymers, especially of polymers suitable for the separati ⁇ on of a ino acids, peptides, proteins and viruses.
  • the aim of the invention was to obtain an improved, water soluble cross-linking agent for use in the preparation of water compatible mechanically stable polymers provided with a special kind of molecular imprint 5 .
  • This special kind of molecular imprinting technique relates to a chromatographic separation method for amino acids, based on ligand-exchange chromatography 6 ("the Davankov method") in combination with the molecular imprinting technique 7 .
  • Water soluble cross- linking agents which can be used in the preparation of polymers containing molecular imprints must have a high degree of water solubility in order to provide for a high degree of cross-linking and consequently a high mechanical stability for chromatographic applications.
  • the structure of the cross-linking agent must be flexible enough to secure the accessability to the active sites in the ready polymer which are provided with molecular imp- rints 8 .
  • the combined ligand-exchange chromatography and molecular imprinting technique is desc ⁇ ribed.
  • a diastereomeric metal ion complex between an amino acid based monomer unit and a selected amino acid is prepared, the complexed amino acid based monomer unit in the metal ion complex is polymerized in the presence of a cross-linking agent (template polyme ⁇ rization) and the selected amino acid is removed from the polymerized metal ion complex to form a molecular imprint of the selected amino acid in the polymeric material.
  • This polymeric material can thereafter be used for chromatogra ⁇ phic (e.g. HPLC) separation of the selected or a structu ⁇ rally similar amino acid from a mixture of other amino acids, inclusive from a racemic mixture of the selected amino acid.
  • the said R'-substituted acryloyl group can also be introduced by reacting a different type of activated acid, for example an active ester between the acid and p-nitrophenol or 1-hydroxybenztriazole, with the heterocyclic compound.
  • the second imino function is acylated with acrylic acid, or preferably with acryloylchloride, to give the compound of the formula I.
  • the acryloyl group can be introduced by using a different type of activated acrylic acid, for example an active ester between acrylic acid and p-nit ⁇ rophenol or 1-hydroxybenztriazole, for the reaction with the intermediate.
  • Both reaction steps are preferably carried out under opti ⁇ mal reaction conditions.
  • an excess of 20 to 40 % of acylating chloride is used compared to the heterocyclic compound II, e.g. piperazine, and the pH of the reaction solution is adjusted and kept between 2.75 and 2.90 and the temperature between 0 and - 5°C when adding the chloride to the reaction solution.
  • the working up of the intermediate e.g. the 1-methacryloylpiperazine, takes place with simple extractions with dichloromethane and ethylacetate or the like.
  • the intermediate, especially the 1-methacryloylpiperazine can in this manner be isolated with high yields (appr. 75 to 80 % of the theor. maximum) and with a very high degree of purity (99 % according to HPLC-analysis) .
  • the second reaction step is carried out in a water/organic solvent two-phase system.
  • the organic solvent can be di ⁇ chloromethane, chloroform, hexane, toluene, benzene or any other inert solvent which is immiscible with water.
  • one equivalent of all reactants e.g. 1-methacryloylpiperazine, acryloylchloride, NaOH
  • the reaction temperature is kept under 0 °C during the addition of acryloylchloride to the reaction solution.
  • the working up of the end product takes place using simple extractions.
  • the end product can in this manner be isolated with high yields (80 to 95 % of the theor. maximum) and with a very high degree of purity (99% according to HPLC- analysis) .
  • the novel cross-linking agents can be polymerized i.a. using the bulk polymerization method, wherein the monomers are first dissolved in a suitable solvent whereby the monomers, during the polymerization, precipitate as a poly- meric material in the solvent.
  • suitable solvents in the bulk polymerization are for example water, methanol, et- hanol, 1-propanol, dichloromethane, chloroform, formic acid, acetic acid, N,N-dimethylformamide, dimethylsulfoxide and acetonitril.
  • the solvent is also called a porogen as it forms pores in the polymeric material.
  • the cross-linking agent can be copolymerized with various functional mono ⁇ mers, i.a. with acryloyl-, methacryloyl-, vinyl-, acrylate- and methacrylate-based monomers.
  • the polymerization can be initiated with thermal or photo- lytic homolysis of azobisnitriles, e.g. 2,2*-azobis(2- methylpropionitrile) [AIBN] .
  • Polymerizations with this cross-linking agent can also be initiated with the peroxo- disulfate/N,N,N' ,N'-tetramethylethylenediamine system commonly used in aqueous mediums.
  • Bead-shaped polymer particles using the novel cross-linking agent can also be prepared using a so-called reversed suspension polymerization method.
  • Such beads can be ob ⁇ tained in varying sizes depending on a variety of factors, such as choice of detergent(s) , volumetric relationsship between water and organic phase, shape of polymerization vessel, stirrer speed, etc.
  • the beads usually of a size of 45 to 80 ⁇ m, have a high mechanical stability and they withstand pressures up to 200 bar without being deformed.
  • the polymers thus obtained using the novel cross-linking agents can be used in a manner described in WO-publication 93/13034, the contents of which is included herein for reference, for the separation of amino acids from a mixture of amino acids, such as for the separation of chiral amino acids from a racemic mixture.
  • the mixture of amino acids to be separated is contac- ted with a polymer material which is composed of cross- linked, amino acid-based monomer units, said polymer meta- rial containing a molecular imprint of the selected amino acid, in which molecular imprint there is also bound a divalent metallic ion and the amino-acid based monomer unit.
  • the amino acid-based monomer is according to one embodiment N-methacrylaminomethyl-L-proline or N-methacryl- aminomethyl-D-proline, which is especially suitable for the preparation of a polymer suitable for the resolution of race ic amino acids.
  • the divalent metal is selected from copper(II) , manganese(II) , iron(II) , cobolt(II) , zinc(II), cadmium(II) and nickel(II), preferably copper(II) .
  • the new cross-linking agents are suitable also for the template polymerization of biological molecules, for example pepti ⁇ des, proteins and viruses.
  • the new cross-linking agent provides a polymeric material having a chemical character compatible with peptides and proteins, and its high mechanical strength and durability makes it suitable for use in chromatography. It is assumed that the novel cross-linking agents possess a suitable degree of flexibi ⁇ lity resulting in a polymeric material having feasible mass transfer properties enabling the diffusion of the large template into and out of the active sites during the chro ⁇ matographic separation process.
  • the porocity and the flexi ⁇ bility of the the polymeric material can be adjusted by the amount and nature of porogen and/or the existence of an extra small inert comonomer during the template polymeriza- tion, or by modifying the degree of cross-linking. Due to their beneficial physico-chemical properties, the novel cross-linking agents, and in particular l-acryloyl-4-met- hacryloyl-piperazine, can be polymerized under different pH-values and temperatures, which is of importance in the template polymerization of proteins and peptides.
  • the chelating monomer can be any monomer containing iminopoly- carboxylate, sulfonate or iminopolyphosphonate groups capable of coordination with the divalent metal ion, capa ⁇ ble of retaining the chelating metal in the resin and possessing a favourable polymerization ratio to the new cross-linking agent.
  • onosubstituted (meth)ac- rylamide based monomers are compatible with the new cross- linking agents, as are the following exemplatory monomers,
  • a common chelating functional group is the iminodiacetate group, e.g. the following new monomer
  • the monomers may be prepared using methods which are analo ⁇ gous to methods already described in literature. (Houben Weil, 8, 4. Aufl. Georg Thieme verlag, 1952, Stuttgart, p. 676; Houben Weil, 10/2, 4. Aufl. Georg Thieme Verlag, 1967, Stuttgart, p. 13; W.T. Read, J. Am. Soc. 36 (1914) 1747-1765)
  • the metals mentioned above may be used as chelating metals.
  • Cu(II) is preferred due to the thermodynamic stability of the mixed Cu(II)-complexes. The net charge on the metal ion must remain positive, or the metal will be stripped from the column.
  • the functional groups co-ordinating with the metal ion the iminopolyphosphonate and iminopolycarboxyla ⁇ te groups hold metal ions better than the sulfonate groups.
  • the size of peptides to be used as templates varies from two amino acids to even some hundred amino acid units.
  • functional groups in the amino acid side chains are capable of co-ordinating to metal ions, such as in histidine and cysteine. Such functional groups make it possible to use metal chelation in the creation of isomer and substrate specific cavities by template polyme ⁇ rization in aqueous solution.
  • the metal should naturally be selected in accordance to the number and types of groups present in the peptides capable of co-ordination with the metal.
  • Favourable water soluble peptides are small di-, tri- and oligopeptides with not too hydrophobic amino acid side chains.
  • Suitable peptides are those formed from e.g. glyci- ne, alanine, serine and threonine.
  • viruses as templates is based on the fact that viruses have proteins on their surfaces, and proteins that contain histidines and cysteines are capable of co-or ⁇ dination e.g. with Cu(II)ions.
  • proteins in general, such as enzymes.
  • the presence of a single histidine residue on the surface of a protein is sufficient for the molecule recognition by specific Cu(II) chelation interaction during the template polymeri ⁇ zation.
  • the protein surface contains several histidine or other chelating groups on its surface, it is recommended to use e.g. Zn(II) , Co(II), Ni(II), Cd(II) or Ca(II) which form thermodynamically less stable complexes with amino acid residues.
  • three polymers were prepared based on the cross-linking agents l-acryloyl-4-methacryloylpipera- zine, 1,4-bisacryloylpiperazine (1,4-bis[l-oxo-2-prope- nyl]-piperazine) and N,N'-methylenebisacrylamide, in order to prepare a polymer which can effectively split amino acid racemates under chromatographic (HPLC) conditions.
  • HPLC chromatographic
  • Example 1 C-D shows that l-acryloyl-4-methacryloylpiperazine gives a highly superior polymer both as regards the racemic separation capability and as regards physical characteris ⁇ tics.
  • the polymer prepared with the commonly used cross- linking agent N,N'-methylenebisacrylamide was much too porous to be tested under HPLC conditions.
  • the solution is slowly brought to room temperature. Thereafter the reaction solution is extracted with 3 x 200 ml dichloromet ⁇ hane. While cooling with water/ice and while vigorously stirring, the reaction solution is saturated with Na 2 C0 3 and the product is extracted thereafter with 4 x 200 ml of ethylacetate. The ethylacetate phases are dried with Na 2 C0 3 . After evaporation and drying in an exsiccator 23.1 g (0.15 moles) of 1-methacryloylpiperazine is obtained as an oil, corresponding to 75 % of the theor. maximum amount.
  • the polymerisation is continued at room temperature in a glass vessel (30 ml) which is provided with a tight fitting plastic lid.
  • the obtained hard polymer is ground, screened in methanol through a 25 ⁇ m screening cloth (Retsch) and sedimented finally in methanol to eliminate undersized particles.
  • the thus obtained material (10-25 ⁇ m) is washed with 1M ammo ⁇ nia, water, 0.1 M CuS0 4 and finally with water.
  • the column containing the polymer is equilibrated in 0.1 M NH 3 with 0.1 mM CuS0 4 in a Kontron HPLC-apparatus.
  • a mixture of D and L-serine (100 ⁇ g D-serine + 100 ⁇ g L-serine) is injected in 20 ⁇ l of mobile phase. Flow: 0.40 ml/min. Counter pressure: 15 bar. Detection at 254 n , separation factor 1.8. Eluation profile, see Figure 1.
  • 1,4-bisacryloylpiperazine is dissolved in 7.0 ml of water. 700 ⁇ l of ammonium peroxodisulfate solution
  • N,N'-methylenebisacrylamide (0.037 moles) of N,N'-methylenebisacrylamide is dissolved in 160 ml of water. 0.125 g of ammonium peroxodisulfate is added, and the solution is purged with nitrogen gas for two minutes. The polymerization is initiated with 40 ⁇ l of Temed. The polymerization is continued at room temperature in a glass vessel which is provided with a tight fitting plastic lid. The obtained soft polymer is washed with 1 M ammonia, water, 0.1 M CuS0 4 and finally with water. The polymer is too soft to be packed in a HPLC-column.
  • EXAMPLE 2 EXAMPLE 2
  • 1,1-dicarboxymet- hyl-2-methacryloylhydrazine is obtained as the product.
  • 2.2 Preparation of a (1, 1-dicarboxymethy1-2-methacryloyl ⁇ hydrazine) -C(II) -(peptide) -complex

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Cette invention concerne de nouveaux agents réticulants du type de la formule (I), dans laquelle X représente: (a), (b), (c), (d), lesquels cycles peuvent comporter 1 à n substituants, n étant égal au nombre d'atomes de carbone protonés de ce cycle et les substituants étant sélectionnés parmi -OH, -SH, -CN, -C(O)NH2, alkyle C1-C3 et hydroxyalkyle C1-C3, ou dans lesdits cycles (a) à (d), au moins un des éléments du cycle -CH2 peut être remplacé par un groupe carbonyle -C(O)-, et R1 est sélectionné dans le groupe comprenant alkyle C¿1?-C3 et hydroxyalkyle C1-C3 et sert à la préparation de polymères dans l'eau ou des solvants organiques. La haute hydrosolubilité de ces agents réticulants par rapport à d'autres les rend propres à la préparation de polymères hydrocompatibles fortement réticulés, utilisables comme matière de séparation chromatographique, de peptides, de protéines, de virus et d'énantiomènes à acides aminés. Le nouvel agent réticulant est synthétisé, par exemple, par l'acylation en deux étapes des fonctions imino du composé hétérocyclique de départ.
PCT/FI1994/000293 1993-06-30 1994-06-28 Nouveaux agents reticulants et leur utilisation WO1995001347A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU70756/94A AU7075694A (en) 1993-06-30 1994-06-28 Novel cross-linking agents and use thereof
EP94919700A EP0706520A1 (fr) 1993-06-30 1994-06-28 Nouveaux agents reticulants et leur utilisation
FI956318A FI956318L (fi) 1993-06-30 1995-12-29 Uudet ristisitojat ja niiden käyttö

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SE9302247A SE9302247L (sv) 1993-06-30 1993-06-30 Ny tvärbindare och dess användning
SE9302247-3 1993-06-30

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0797096A3 (fr) * 1996-03-20 1998-01-07 Bio-Rad Laboratories, Inc. Reconnaissance sélective des solutés dans un milieux chromatographique par affinité créée artificiellement
US5756717A (en) * 1995-05-24 1998-05-26 Perseptive Biosystems, Inc Protein imaging
WO2019002535A1 (fr) * 2017-06-29 2019-01-03 Mipsalus Aps Polymères à empreinte moléculaire ciblant la phénylalanine
US10633418B2 (en) 2011-03-09 2020-04-28 Jitsubo Co., Ltd. Cross-linked peptides containing non-peptide cross-linked structure, method for synthesizing cross-linked peptides, and novel organic compound used in method
US10654891B2 (en) 2013-09-09 2020-05-19 Jitsubo Co., Ltd. Cross-linked peptides containing non-peptide cross-linked structure, method for synthesizing cross-linked peptides, and novel organic compound used in method

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GB1218780A (en) * 1967-12-28 1971-01-13 Matsushita Electric Ind Co Ltd Dry cells

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GB1218780A (en) * 1967-12-28 1971-01-13 Matsushita Electric Ind Co Ltd Dry cells

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Title
ANALYTICAL BIOCHEMISTRY, Volume 173, 1988, DENIS F. HOCHSTRASSER et al., "Development of Polyacrylamide Gels that Improve the Separation of Proteins and Their Detection by Silver Staining", page 412 - page 423. *
CHEMICAL ABSTRACTS, Vol. 68, 1968, (Columbus, Ohio, USA), SHIGEHO INABA et al., "Substituted piperazines", page 4815, THE ABSTRACT No. 49649M; & JP,B,42 011 955. *
CHEMICAL ABSTRACTS, Vol. 69, 1968, (Columbus, Ohio, USA), KINOSHITA YOSHIRO et al., "alpha, beta - Unsaturated monocarboxylic piperazides", page 6301 THE ABSTRACT No. 67423h; JP,B,42 024 912. *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5756717A (en) * 1995-05-24 1998-05-26 Perseptive Biosystems, Inc Protein imaging
EP0797096A3 (fr) * 1996-03-20 1998-01-07 Bio-Rad Laboratories, Inc. Reconnaissance sélective des solutés dans un milieux chromatographique par affinité créée artificiellement
US5814223A (en) * 1996-03-20 1998-09-29 Bio-Rad Laboratories, Inc. Selective recognition of solutes in chromatographic media by artificially created affinity
US5916445A (en) * 1996-03-20 1999-06-29 Bio-Rad Laboratories, Inc. Selective recognition of solutes in chromatographic media by artificially created affinity
US10633418B2 (en) 2011-03-09 2020-04-28 Jitsubo Co., Ltd. Cross-linked peptides containing non-peptide cross-linked structure, method for synthesizing cross-linked peptides, and novel organic compound used in method
US10654891B2 (en) 2013-09-09 2020-05-19 Jitsubo Co., Ltd. Cross-linked peptides containing non-peptide cross-linked structure, method for synthesizing cross-linked peptides, and novel organic compound used in method
WO2019002535A1 (fr) * 2017-06-29 2019-01-03 Mipsalus Aps Polymères à empreinte moléculaire ciblant la phénylalanine
JP2020525605A (ja) * 2017-06-29 2020-08-27 ミプサルス エーピーエス フェニルアラニンを標的とする分子インプリントポリマー
US11028203B2 (en) 2017-06-29 2021-06-08 Mipsalus Aps Molecular imprinted polymers targeting phenylalanine
US11879027B2 (en) 2017-06-29 2024-01-23 Mipsalus Aps Molecular imprinted polymers targeting phenylalanine

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SE9302247L (sv) 1994-12-31
EP0706520A1 (fr) 1996-04-17
AU7075694A (en) 1995-01-24
SE9302247D0 (sv) 1993-06-29

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