Affinity chromatography-based method for measuring supramolecular interaction dissociation rate constant
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A dissociation rate constant and supramolecular technology, applied in the field of supramolecular interactions, can solve problems such as weak interactions
Inactive Publication Date: 2015-05-20
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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U.S. Patent US20080293587 discloses a biological target screening method based on affinity chromatography with transient weak interactions between ligands and receptors, mainly focusing on
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Embodiment 1
[0106] Embodiment 1: The present invention is used for measuring the dissociation rate constant of acetaminophen-β-cyclodextrin supramolecule
[0107] The method and result of table 1 embodiment 1
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[0110] test methods:
[0111] (1) Provide an affinity chromatography column bonded with the target main molecule;
[0112] (2) Prepare the mobile phase and use the mobile phase to prepare the sample solution;
[0113] (3) At 25°C, inject 5 μL of the sample solution into the chromatographic system at the first predetermined flow rate in Zonal elution (Agilent binary pump G1311C, Agilent autosampler G1329B; Agilent diode array detector G4212B, Agilent column Incubator G1316A, Agilent ChemStation Chromatographic Workstation), and then use the mobile phase to elute under the flow rate conditions;
[0114] (4) Collect the signal-time curve of the first guest molecule to be analyzed on the β-cyclodextrin affinity chromatographic column, that is, the chromatogra...
Embodiment 2
[0122] Embodiment 2: The present invention is used for determining the dissociation rate constant of sertraline-β-cyclodextrin supramolecule
[0123] The dissociation rate constant of the sertraline-β-cyclodextrin supramolecule was measured by the same method as in Example 1, and the measurement results are shown in Table 2 below.
[0124] The method and result of table 2 embodiment 2
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Embodiment 3
[0128] Embodiment 3: The present invention is used for the simultaneous determination of dissociation rate constants of acetaminophen, phenacetin and flurbiprofen-β-cyclodextrin supramolecules
[0129] The dissociation rate constants of acetaminophen, phenacetin and flurbiprofen-β-cyclodextrin supramolecules were measured by the same method as in Example 1, and the results are shown in Table 3 below.
[0130] The method and result of table 3 embodiment 3
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Abstract
The invention relates to an affinity chromatography-based method for measuring a dissociation rate constant, and especially aims at relatively extensive and relatively weak supramolecular interaction. The affinity chromatography-based method is a method for firstly fixing a host molecule and then measuring the host molecule, and comprises the following steps: under a certain temperature condition and at various flow rates, respectively acquiring chromatography flow curves of a first to-be-analyzed guest molecule and a second to-be-analyzed guest molecule on an affinity chromatography column which is bonded with a target host molecule, calculating retention time, half peak width and tower plate height by peak fitting, and calculating the dissociation rate constant. According to the method disclosed by the invention, special apparatus and equipment are not needed, and a normal chromatographic analysis method is adopted, so that the dynamic speed constant of the supramolecular weak interaction can be measured; meanwhile, the method can be combined with various detectors of ultraviolet rays, fluorescence, mass spectrum and the like to achieve unique advantages in detection of low-concentration drugs with poor water solubility as well as low-cost and quick measurement of supramolecular interaction dynamic parameters.
Description
technical field [0001] The invention relates to a method for measuring dissociation rate constants based on affinity chromatography, especially for weaker and wider supramolecular interactions. Background technique [0002] Supramolecules are generally aggregates of two or more molecules. The supramolecules formed by drugs and excipients are usually formed by non-covalent interactions between host molecules (supramolecular carriers) and guest molecules (drugs) through hydrogen bonds and van der Waals forces. molecular complex. Drugs and supramolecular carriers, such as cyclodextrins, crown ethers, and dendrimers, form supramolecular complexes that can improve the water solubility, bioavailability, and targeting of drugs, and play an important role in anti-tumor, anti-inflammatory and other fields. play an important role. [0003] The interaction between host and guest molecules, that is, host-guest recognition, is the basis for supramolecules to exert their special functio...
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IPC IPC(8): G01N30/88
Inventor 李海燕葛静文殷宪振孙立新张继稳
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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