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WO2015098152A1 - Procédé pour préparer un échantillon de cristallographie et procédé pour déterminer la structure moléculaire de métabolite - Google Patents

Procédé pour préparer un échantillon de cristallographie et procédé pour déterminer la structure moléculaire de métabolite Download PDF

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WO2015098152A1
WO2015098152A1 PCT/JP2014/067233 JP2014067233W WO2015098152A1 WO 2015098152 A1 WO2015098152 A1 WO 2015098152A1 JP 2014067233 W JP2014067233 W JP 2014067233W WO 2015098152 A1 WO2015098152 A1 WO 2015098152A1
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metabolite
sample
structure analysis
crystal structure
crystal
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Japanese (ja)
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藤田　誠
泰英 猪熊
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Japan Science and Technology Agency
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
    • G01N23/20—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
    • G01N23/207—Diffractometry using detectors, e.g. using a probe in a central position and one or more displaceable detectors in circumferential positions
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J1/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
    • C07J1/0003—Androstane derivatives
    • C07J1/0011—Androstane derivatives substituted in position 17 by a keto group
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J1/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
    • C07J1/0003—Androstane derivatives
    • C07J1/0018—Androstane derivatives substituted in position 17 beta, not substituted in position 17 alfa
    • C07J1/0022—Androstane derivatives substituted in position 17 beta, not substituted in position 17 alfa the substituent being an OH group free esterified or etherified

Definitions

  • the present invention relates to a method for preparing a crystal structure analysis sample useful in determining the molecular structure of a metabolite, and to determine the molecular structure of a metabolite using the crystal structure analysis sample obtained by this method. Regarding the method.
  • Patent Document 1 discloses a method for analyzing a drug metabolite, which combines RI (radioisotope) quantitative analysis and mass spectrometry.
  • an X-ray crystal structure analysis method is known as a method for determining the molecular structure of an organic compound. This is an extremely useful method because it can be solved by an X-ray crystal structure.
  • it is necessary to repeat a number of trial and error experiments until finding the optimum conditions for crystallization. It was necessary to secure a sample of several milligrams.
  • the molecular structure of the trace amount substance is determined by the X-ray crystal structure analysis method, it is necessary to repeat a kinetic experiment or perform another chemical synthesis in order to secure a sample of several milligrams.
  • metabolites such as drug metabolites often have large differences in physical properties between the parent compound and the unknown metabolite, and there are a wide variety of compounds, and there are many, so the crystallization conditions for each compound A great deal of time and money is spent on optimizing.
  • the molecular structure of metabolites often changes in various ways depending on pH, salt concentration, etc., and it is difficult to crystallize unless it is chemically pure, including the molecular structure (conformation).
  • the present invention has been made in view of such circumstances, a method for preparing a crystal structure analysis sample useful for determining the molecular structure of a metabolite, and a crystal structure analysis sample obtained by this method It is an object to provide a method for determining the molecular structure of a metabolite using
  • a target crystal structure analysis sample can be prepared by bringing a solvent solution containing a metabolite into contact with a single crystal of a specific porous compound.
  • the headline and the present invention have been completed.
  • a method for preparing a crystal structure analysis sample for determining the molecular structure of a metabolite, Three-dimensional skeleton composed of one or two or more molecular chains, or one or two or more molecular chains and a skeleton-forming compound, and three-dimensionally regularly formed by being partitioned by the three-dimensional skeleton Contacting a single crystal of a porous compound having pores and / or cavities with a solvent solution containing the metabolite, Preparation of a crystal structure analysis sample, comprising a step of preparing a crystal structure analysis sample in which the metabolite molecules are regularly arranged in the pores and / or hollows of the single crystal Method.
  • the porous compound is a polynuclear metal complex formed in a self-organizing manner from a plurality of ligands having two or more coordination sites and a plurality of metal ions as a central metal. 5.
  • the ligand having two or more coordination sites is represented by the following formula (1):
  • Ar represents a trivalent aromatic group which may have a substituent.
  • X1 to X3 each independently represent a divalent organic group or Ar and Y1 to Y3 directly.
  • Y1 to Y3 each independently represents a monovalent organic group having a coordination site.
  • a method for determining the molecular structure of a metabolite comprising crystal structure analysis data for a crystal structure analysis sample obtained by the method according to any one of (1) to (9), and a parent of the metabolite
  • a method for preparing a crystal structure analysis sample useful for determining a molecular structure of a metabolite, and a molecular structure of a metabolite using a crystal structure analysis sample obtained by this method are obtained.
  • a method for determining is provided.
  • a sample for crystal structure analysis can be prepared even if the metabolite whose molecular structure is to be determined is a substance that is difficult to obtain a single crystal, or a liquid or gaseous substance at room temperature.
  • the molecular structure of the metabolite can be determined.
  • a crystal structure analysis sample can be prepared and the molecular structure of the metabolite can be determined.
  • FIG. 1 is a diagram illustrating a three-dimensional skeleton of a polynuclear metal complex 1.
  • FIG. 3 is a diagram illustrating a three-dimensional network structure of a polynuclear metal complex 3.
  • FIG. 3 is a diagram illustrating a three-dimensional network structure of a polynuclear metal complex 5.
  • FIG. 1 is a diagram showing a polynuclear metal complex obtained by inclusion of an adrenosterone metabolite obtained in Example 1.
  • FIG. It is a figure showing the molecular structure of an adrenosterone metabolite.
  • FIG. 2 is a diagram showing a polynuclear metal complex obtained by inclusion of an ethyl 2-oxocyclohexanecarboxylate metabolite obtained in Example 2.
  • FIG. 2 is a diagram showing the molecular structure of an ethyl 2-oxocyclohexanecarboxylate metabolite.
  • 4 is a diagram showing a polynuclear metal complex obtained by inclusion of a testosterone metabolite obtained in Example 3.
  • FIG. It is a figure showing the molecular structure of a testosterone metabolite.
  • FIG. 3 is a diagram showing a polynuclear metal complex obtained by inclusion of methyl 5-methoxy-2-oxo-1,2,3,4-tetrahydronaphthalene-1-carboxylate metabolite obtained in Example 4.
  • FIG. 3 is a diagram showing the molecular structure of a methyl-5-methoxy-2-oxo-1,2,3,4-tetrahydronaphthalene-1-carboxylate metabolite.
  • FIG. 6 is a view showing a polynuclear metal complex obtained by inclusion of a benzyl metabolite obtained in Example 5. It is a figure showing the molecular structure of a benzyl metabolite.
  • FIG. 6 is a diagram showing a polynuclear metal complex obtained by inclusion of a DDT metabolite obtained in Example 6.
  • FIG. It is a figure showing the molecular structure of a DDT metabolite.
  • FIG. 6 is a diagram showing a polynuclear metal complex obtained by inclusion of adrenosterone obtained in Example 7. It is a figure showing the molecular structure of adrenosterone. It is a comparison figure of the position in the crystal
  • the method for preparing a crystal structure analysis sample of the present invention is a method for preparing a crystal structure analysis sample for determining the molecular structure of a metabolite, and includes one or more samples.
  • a three-dimensional skeleton composed of molecular chains or one or more molecular chains and a skeleton-forming compound, and three-dimensionally regularly arranged pores formed by partitioning the three-dimensional skeleton and / or Or by contacting a single crystal of a porous compound having a hollow with a solvent solution containing the metabolite so that the metabolite molecules are regularly placed in the pores and / or hollows of the single crystal. It has the process of producing the sample for crystal structure analysis arranged.
  • the metabolite targeted by the present invention is an intermediate product or final product of a metabolic process.
  • the type of metabolite is not particularly limited. Examples thereof include metabolites of pharmaceutically active ingredients, metabolites of pesticidal active ingredients, food metabolites, and the like. Among these, since it is expected that the development period of a drug superior in efficacy and safety can be greatly shortened, in the present invention, a metabolite of a pharmaceutically active ingredient and a metabolite of an agrochemical active ingredient are preferable, and the drug activity Component metabolites are particularly preferred.
  • reaction in the first phase reactions that lower the molecular weight of the target substance (decomposition) or do not change significantly
  • reactions that add other molecules increase the molecular weight
  • reaction in the second phase also called conjugation.
  • the first phase reaction examples include an oxidation reaction, a hydroxylation reaction, a hydrolysis reaction, a reduction reaction, and a hydration reaction.
  • molecules added in the second phase reaction include sulfuric acid, acetic acid, glutathione, and glucuronic acid.
  • metabolites targeted by the present invention metabolites generated by the first phase reaction, that is, the parent compound, are obtained by oxidation reaction, hydroxylation reaction, hydrolysis reaction, reduction reaction, hydration reaction. Modified compounds are preferred.
  • the metabolite targeted by the present invention is preferably a compound whose chemical change from the parent compound is within 30% of the total chemical bond of the parent compound, more preferably a compound within 25%. Preferably, it is more preferably within 20%.
  • the “chemically changed part from the parent compound” means a part of all the carbon atoms of the parent compound that has changed its binding environment (in a chemical bond such as a covalent bond or a coordinate bond, it becomes a partner of the bond) The part where the atom or atomic group changed before and after metabolism, or the part where the bond order changed).
  • adrenosterone has 19 carbon atoms in the molecule
  • adreosterone metabolites are compounds in which the binding environment of 3 carbon atoms out of the 19 carbon atoms of the parent compound is changed. is there.
  • the portion chemically changed from the parent compound is 15.79% of the total chemical bond of the parent compound.
  • the number of “parts chemically changed from the parent compound” is preferably 5 or less, more preferably 4 or less, and even more preferably 3 or less.
  • changes in the structure of the parent compound and metabolite include changes in the rotation angle of substituents around a rotatable single bond and variable conformations, as in the examples shown below. Changes may be observed. These are not included in chemical changes.
  • the obtained metabolite is a mixture of those having different substituent rotation angles, bond angles, conformations, etc., or different solvation states (ie, isomer mixtures, substituent rotations). Even a mixture of compounds having different angles, bond angles, conformations, etc.), these can be included in the hollow interior with regularity, and each isomer, rotation angle of substituent, bond angle, It is also possible to analyze the molecular structure of a plurality of compounds having different conformations.
  • the size of the metabolite is not particularly limited as long as the metabolite is large enough to enter the pores and / or hollows of the single crystal.
  • the molecular weight of the metabolite is usually 20 to 3,000, preferably 100 to 2,000.
  • the molecular size of the metabolite is grasped to some extent by nuclear magnetic resonance spectroscopy, mass spectrometry, elemental analysis, etc., and a single crystal having appropriate pores and hollows is appropriately selected and used in advance. It is also preferable.
  • the solvent of the solvent solution containing the metabolite is not particularly limited as long as it does not dissolve the single crystal to be used and dissolves the metabolite.
  • the solvent solution containing a metabolite may be one in which the metabolite is in an equilibrium mixture state in the solution.
  • a plurality of isomers of the equilibrium mixture are included.
  • An in contact porous metal complex can be obtained.
  • the crystal structure analysis method of the present invention when compounds having different molecular structures (including compounds having different conformations) are included in the hollow or pores, peaks derived from the respective compounds are observed. As a result, it is possible to analyze the molecular structure of all compounds.
  • the solvent to be used is normal pressure (1 ⁇ 10 5 Pa).
  • the boiling point is preferably 200 ° C. or less, more preferably ⁇ 50 to + 185 ° C., and further preferably 30 to 80 ° C.
  • solvent used include aromatic hydrocarbons such as benzene, toluene, xylene, chlorobenzene, 1,2-dichlorobenzene and nitrobenzene; fats such as n-butane, n-pentane, n-hexane and n-heptane.
  • aromatic hydrocarbons such as benzene, toluene, xylene, chlorobenzene, 1,2-dichlorobenzene and nitrobenzene
  • fats such as n-butane, n-pentane, n-hexane and n-heptane.
  • Aromatic hydrocarbons such as cyclopentane, cyclohexane and cycloheptane; Nitriles such as acetonitrile and benzonitrile; Sulfoxides such as dimethyl sulfoxide (DMSO); N, N-dimethylformamide and n-methyl Amides such as pyrrolidone; Ethers such as diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane and 1,4-dioxane; Alcohols such as methanol, ethanol and isopropyl alcohol; Ketones such as acetone, methyl ethyl ketone and cyclohexanone; Cellosolves such as lucerosolv; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, and 1,2-dichloroethane; esters such as methyl acetate, ethyl acetate, ethyl lac
  • the content of the metabolite in the solvent solution containing the metabolite is preferably 5 mg or less, more preferably 0.5 ⁇ g to 1 mg, and further preferably 1 ⁇ g to 0.5 mg.
  • the preparation method of the present invention can efficiently prepare a sample for crystal structure analysis even when a metabolite can be obtained only in a trace amount of 5 mg or less. When the amount of metabolites exceeds 5 mg, it is considered that there is a high possibility that a single crystal can be obtained because the crystallization conditions can be sufficiently studied even when a conventional method for producing a single crystal is used. .
  • the method for preparing a sample for crystal structure analysis of the present invention is particularly useful when the content of metabolites is 5 mg or less, where it has been difficult to obtain a single crystal by the conventional crystallization method.
  • the concentration of the metabolite in the solvent solution containing the metabolite is not particularly limited, but is usually 0.001 to 50 ⁇ g / ⁇ L, preferably 0.01 from the viewpoint of efficiently producing a high-quality crystal structure analysis sample. ⁇ 5 ⁇ g / ⁇ L, more preferably 0.1 to 1 ⁇ g / ⁇ L.
  • the solvent solution containing the metabolite is a biological sample solution collected from a living body, a culture medium, and a purification treatment liquid obtained by performing a purification treatment of the biological sample solution or the culture medium. (Hereinafter sometimes referred to as “purified treatment solution”).
  • Bio sample fluids collected from living bodies are obtained by treating body fluids such as blood (plasma), urine, sweat, saliva, bile; cells, fungi (microorganisms), tissues, organs, etc. Homogenate solution; and the like.
  • body fluids such as blood (plasma), urine, sweat, saliva, bile; cells, fungi (microorganisms), tissues, organs, etc. Homogenate solution; and the like.
  • the medium examples include a medium after culturing tissues, cells, and fungi (microorganisms). These media may be either liquid media or solid media. In the case of a solid medium, it can be used for the method of the present invention by preparing a solution by performing a solvent extraction treatment described later.
  • the purification method is not particularly limited. Examples include purification methods such as centrifugation, filtration, dialysis, solvent extraction, electrophoresis, and liquid chromatography. These purification methods can be used singly or in combination of two or more.
  • purification treatment liquid an extraction liquid after solvent extraction, an eluent after liquid chromatography, or a solution in which the concentration of these liquids is adjusted is preferable because it can be obtained efficiently.
  • a single crystal of a porous compound used in the present invention has a three-dimensional skeleton and a three-dimensionally regularly arranged fine particle formed by partitioning with the three-dimensional skeleton. It has a hole and / or a hollow. If there is a three-dimensional skeleton and three-dimensionally regularly arranged pores and / or hollows that are partitioned by the three-dimensional skeleton, the pores and / or hollows.
  • the metabolite to be subjected to the crystal structure analysis can be regularly (with a certain regularity) and included, and as a result, the crystal structure can be analyzed using the inclusion.
  • the three-dimensional skeleton refers to a skeleton-like structure having a three-dimensional extension inside a single crystal.
  • the three-dimensional skeleton is composed of one or more molecular chains, or one or two or more molecular chains and a skeleton-forming compound.
  • “Molecular chain” refers to an organization organized by covalent bonds and / or coordinate bonds. This molecular chain may have a branched structure or a cyclic structure. Examples of the three-dimensional skeleton composed of one molecular chain include a skeleton organized in a “jungle gym” shape.
  • a three-dimensional skeleton composed of two or more molecular chains
  • two or more molecular chains are organized as a whole by interactions such as hydrogen bonds, ⁇ - ⁇ stacking interactions, van der Waals forces, etc.
  • the skeleton For example, there is a skeleton in which two molecular chains are entangled in a “Chienowa” shape.
  • Examples of such a three-dimensional skeleton include the three-dimensional skeletons of polynuclear metal complexes 1 and 2 described later.
  • “Skeletogenic compounds” do not constitute part of the molecular chain, but constitute part of the three-dimensional skeleton by interactions such as hydrogen bonds, ⁇ - ⁇ stacking interactions, van der Waals forces, etc. Refers to the compound.
  • the skeleton-forming aromatic compound in the polynuclear metal complex mentioned later is mentioned.
  • “Three-dimensionally ordered pores and / or hollows” means pores and hollows that are regularly aligned without being disturbed to the extent that pores and hollows can be confirmed by crystal structure analysis.
  • Pore and “hollow” represent an internal space in the single crystal. The internal space extending in a cylindrical shape is called “pore”, and the other internal space is called “hollow”.
  • the size of the pore is defined as an inscribed circle of the pore (hereinafter simply referred to as a parallel plane) parallel to the crystal plane that is closest to the perpendicular to the direction in which the pore extends (hereinafter simply referred to as a parallel plane).
  • a parallel plane parallel to the crystal plane that is closest to the perpendicular to the direction in which the pore extends
  • the “direction in which the pores extend” can be determined by the following method. That is, first, a crystal plane X (A plane, B plane, C plane, or a diagonal plane of each) in an appropriate direction across the target pore is selected. Then, by expressing the atoms that exist on the crystal plane X and constitute the three-dimensional skeleton using the van der Waals radii, a cross-sectional view of the pore having the crystal plane X as a cutting plane is drawn. Similarly, a cross-sectional view of a pore having a crystal plane Y shifted from the crystal plane X by one unit cell as a cut plane is drawn.
  • the centers of the cross-sectional shapes of the pores in the respective crystal planes are connected with a straight line (dashed line) in the three-dimensional view (see FIG. 1).
  • the direction of the straight line obtained at this time is the direction in which the pores extend.
  • the “diameter of the inscribed circle of the pore” can be obtained by the following method. That is, first, a cross-sectional view of the pore having the parallel plane as a cut plane is drawn by the same method as described above. Next, after drawing the inscribed circle of the pore in the cross-sectional view and measuring the diameter, the obtained measured value is converted into an actual scale to obtain the diameter of the inscribed circle of the actual pore. be able to. Furthermore, by measuring the diameter of the inscribed circle of the pore in each parallel surface while gradually translating the parallel surface by one unit cell, the diameter of the inscribed circle of the narrowest part and the widest The diameter of the inscribed circle of the part is obtained.
  • the diameter of the inscribed circle of the single crystal pores used in the present invention is preferably 2 to 30 mm, and more preferably 3 to 10 mm.
  • the major axis of the inscribed ellipse of the single crystal pores used in the present invention is preferably 2 to 30 mm, and more preferably 3 to 10 mm.
  • the minor axis of the inscribed ellipse of the single crystal pores is preferably 2 to 30 mm, and more preferably 3 to 10 mm.
  • the pore volume of the single crystal used in the present invention is described in the article Acta Crystallogr. A 46, 194-201 (1990). That is, it is possible to calculate using “volume of single crystal ⁇ porosity in unit cell” based on Solvent Accessible Void (void volume in a unit cell) calculated by a calculation program (PLATON SQUEEZE PROGRAM).
  • Single crystals of pore volume used in the present invention (all pore volume in the grain of the single crystal) is preferably 1 ⁇ 10 -7 ⁇ 0.1mm 3, 1 ⁇ 10 -5 ⁇ 1 ⁇ 10 - 3 mm 3 is more preferable.
  • the size of the hollow is also the same as that of the article Acta Crystallogr. A, 46, 194-201 (1990).
  • the single crystal used in the present invention preferably has a cubic or cuboid shape. One side thereof is preferably 10 to 1000 ⁇ m, more preferably 60 to 200 ⁇ m. By using a single crystal having such a shape and size, a good quality crystal structure analysis sample can be easily obtained.
  • the single crystal to be used is irradiated with MoK ⁇ rays (wavelength: 0.71 ⁇ ) generated at a tube voltage of 24 kV and a tube current of 50 mA, and when diffracted X-rays are detected by a CCD detector, at least 1.5 ⁇ . Those that can determine the molecular structure with a resolution of 1 are preferred. By using a single crystal having such characteristics, a sample for crystal structure analysis of good quality can be easily obtained.
  • the single crystal of the porous compound is not particularly limited as long as it has the above pores and / or hollows.
  • a single crystal of a polynuclear metal complex, a urea crystal, or the like can be given.
  • a crystal of a polynuclear metal complex is preferable because it can easily control the size of pores and hollows and the environment (polarity and the like) in the pores and hollows.
  • polynuclear metal complex examples include a plurality of ligands having two or more coordination sites and a plurality of metal ions as a central metal.
  • the ligand having two or more coordinating sites (hereinafter sometimes referred to as “polydentate ligand”) is not particularly limited as long as it can form the three-dimensional skeleton. Multidentate ligands can be utilized.
  • the “coordinating moiety” refers to an atom or atomic group in a ligand having an unshared electron pair capable of coordinating bond. Examples thereof include heteroatoms such as nitrogen atom, oxygen atom, sulfur atom and phosphorus atom; atomic groups such as nitro group, amino group, cyano group and carboxyl group; Especially, the atomic group containing a nitrogen atom or a nitrogen atom is preferable.
  • a multidentate ligand with a long distance from the center of the ligand to the coordination site a single crystal of a polynuclear metal complex having relatively large pores and hollows is obtained.
  • a multidentate ligand having a short distance from the center of the child to the coordination site a single crystal of a polynuclear metal complex having relatively small pores and hollows can be obtained.
  • the polydentate ligand is preferably a polydentate ligand having two or more coordination sites. More preferred is a ligand having three functional sites (hereinafter sometimes referred to as a “tridentate ligand”), and the unshared electron pairs (orbitals) of the three coordinating sites exist on a quasi-coplanar surface. In addition, it is more preferable that the three coordinating sites are arranged radially at equal intervals with respect to the center portion of the tridentate ligand.
  • each unshared electron pair is on the same plane or is slightly displaced from the plane, for example, 20 ° or less with respect to the reference plane. It also includes the state that exists in a plane that intersects at.
  • three coordinating sites are arranged radially at equal intervals with respect to the central portion of the tridentate ligand means that on a line extending radially from the central portion of the ligand at equal intervals, It means a state in which three coordination sites are arranged at approximately the same distance from the central portion.
  • tridentate ligand for example, the following formula (1)
  • Ar represents a trivalent aromatic group which may have a substituent.
  • X1 to X3 are each independently a divalent organic group, or Ar and Y1 to Y3 are directly connected.
  • Y1 to Y3 each independently represents a monovalent organic group having a coordinating moiety, and a ligand represented by the following formula:
  • Ar represents a trivalent aromatic group.
  • the number of carbon atoms constituting Ar is usually 3 to 22, preferably 3 to 13, and more preferably 3 to 6.
  • Ar is a trivalent aromatic group having a monocyclic structure composed of one 6-membered aromatic ring or a trivalent aromatic group having a condensed ring structure formed by condensing three 6-membered aromatic rings. Groups.
  • Examples of the trivalent aromatic group having a monocyclic structure composed of one 6-membered aromatic ring include groups represented by the following formulas (2a) to (2d).
  • Examples of the trivalent aromatic group having a condensed ring structure formed by condensing three 6-membered aromatic rings include groups represented by the following formula (2e).
  • “*” represents a bonding position with X1 to X3, respectively.
  • Ar may have a substituent at an arbitrary position of the aromatic group represented by formula (2a), formula (2c) to formula (2e).
  • substituents include alkyl groups such as methyl, ethyl, isopropyl, n-propyl, and t-butyl; alkoxy groups such as methoxy, ethoxy, n-propoxy, and n-butoxy; fluorine A halogen atom such as an atom, a chlorine atom or a bromine atom;
  • an aromatic group represented by the formula (2a) or (2b) is preferable, and an aromatic group represented by the formula (2b) is particularly preferable.
  • X1 to X3 each independently represent a divalent organic group or a single bond directly connecting Ar and Y1 to Y3.
  • the divalent organic group those capable of forming a ⁇ -electron conjugated system together with Ar are preferable. Since the divalent organic group represented by X1 to X3 constitutes a ⁇ -electron conjugated system, the planarity of the tridentate ligand represented by the formula (1) is improved, and a stronger three-dimensional network structure is obtained. It becomes easy to form.
  • the number of carbon atoms constituting the divalent organic group is preferably 2 to 18, more preferably 2 to 12, and further preferably 2 to 6.
  • divalent organic group examples include a divalent unsaturated aliphatic group having 2 to 10 carbon atoms, a divalent organic group having a monocyclic structure consisting of one 6-membered aromatic ring, and 2 to 4 6-membered aromatic rings.
  • Examples of the divalent unsaturated aliphatic group having 2 to 10 carbon atoms include vinylene group and acetylene group (ethynylene group).
  • Examples of the divalent organic group having a monocyclic structure composed of one 6-membered aromatic ring include a 1,4-phenylene group.
  • Examples of the divalent organic group having a condensed ring structure in which 2 to 4 6-membered aromatic rings are condensed include a 1,4-naphthylene group and an anthracene-1,4-diyl group. Examples of combinations of two or more of these divalent organic groups include the following.
  • divalent organic groups may have a substituent.
  • substituent include the same as those described above as the substituent for Ar.
  • the divalent organic groups represented by X1 to X3 are preferably the following.
  • Y1 to Y3 each independently represent a monovalent organic group having a coordination site.
  • the organic group represented by Y1 to Y3 those capable of forming a ⁇ -electron conjugated system together with Ar and X1 to X3 are preferable.
  • the organic groups represented by Y1 to Y3 form a ⁇ -electron conjugated system, the planarity of the tridentate ligand represented by the formula (1) is improved, and a strong three-dimensional network structure is easily formed.
  • the number of carbon atoms constituting Y1 to Y3 is preferably 5 to 11, and more preferably 5 to 7.
  • Examples of Y1 to Y3 include organic groups represented by the following formulas (3a) to (3f).
  • “*” represents a bonding position with X1 to X3.
  • Y1 to Y3 may have a substituent at any position of the organic group represented by the formulas (3a) to (3f).
  • substituents include the same as those exemplified above as the substituent for Ar.
  • the group represented by the formula (3a) is particularly preferable.
  • the size of the single crystal pores and hollows can be adjusted.
  • a single crystal having pores and hollows with a size capable of including the target metabolite can be efficiently obtained.
  • tridentate ligand represented by the formula (1) since a strong three-dimensional network structure is easily formed, the planarity and symmetry are high, and the ⁇ -conjugated system spreads throughout the ligand. Those are preferred.
  • tridentate ligands include ligands represented by the following formulas (4a) to (4f).
  • the tridentate ligand represented by the formula (1) includes 2,4,6-tris (4-pyridyl) -1,3,5-triazine (TPT) represented by the above formula (4a). Is particularly preferred.
  • a commercial item can also be used as a polydentate ligand of a polynuclear metal complex.
  • PCP Coordination Polymer
  • MOF Metal Organic Structure
  • the metal ion as the central metal of the polynuclear metal complex is not particularly limited as long as it can form a coordinate bond with the polydentate ligand to form a three-dimensional skeleton.
  • ions of metals in Group 8 to 12 of the periodic table such as iron ions, cobalt ions, nickel ions, copper ions, zinc ions, silver ions, palladium ions, ruthenium ions, rhodium ions, platinum ions, etc. are preferable.
  • metal ions of Groups 8 to 12 of the periodic table are more preferred.
  • zinc (II) ions and cobalt (II) ions are preferred because single crystals having large pores and hollows are easily obtained.
  • a monodentate ligand may be coordinated with the central metal of the polynuclear metal complex.
  • Such monodentate ligands include monovalent anions such as chloride ion (Cl-), bromide ion (Br-), iodide ion (I-), thiocyanate ion (SCN-); ammonia, monoalkyl Electrically neutral coordinating compounds such as amine, dialkylamine, trialkylamine, and ethylenediamine; and the like.
  • the polynuclear metal complex is a reaction solvent (the solvent used for the synthesis of the polynuclear metal complex), a substitution solvent (refers to another solvent replaced with the reaction solvent, the same applies hereinafter), and a skeleton-forming aromatic described later. It may contain a compound.
  • “Skelet-forming aromatic compound” means an aromatic compound that interacts with a molecular chain constituting a three-dimensional skeleton (excluding covalent bonds and coordinate bonds) and can constitute a part of the three-dimensional skeleton.
  • the polynuclear metal complex contains a skeleton-forming aromatic compound, the three-dimensional skeleton tends to become stronger, and the three-dimensional skeleton is more stable even after inclusion of metabolite molecules that determine the molecular structure. There is a case.
  • Examples of the skeleton-forming aromatic compound include condensed polycyclic aromatic compounds. Examples thereof include those represented by the following formulas (5a) to (5i).
  • polynuclear metal complex examples include the following compounds. (1) Compound consisting only of ligand and metal ion [polynuclear metal complex ( ⁇ )] (2) Compound [polynuclear metal complex ( ⁇ )] comprising the polynuclear metal complex ( ⁇ ) and a skeleton-forming aromatic compound (3) A compound in which a guest molecule such as a solvent molecule is included in the polynuclear metal complex ( ⁇ ) or polynuclear metal complex ( ⁇ ) [polynuclear metal complex ( ⁇ )]. Of these polynuclear metal complexes, the polynuclear metal complex ( ⁇ ) and the polynuclear metal complex ( ⁇ ) are sometimes referred to as “host molecules”.
  • the polynuclear metal complex used in the present invention does not lose crystallinity even after incorporating a metabolite molecule that determines the molecular structure into the pore or hollow, and has a relatively large pore or hollow. preferable.
  • the polynuclear metal complex having such characteristics can be easily obtained by using the tridentate ligand represented by the formula (1).
  • Examples of the polynuclear metal complex obtained by using the tridentate ligand represented by the formula (1) include polynuclear metal complexes represented by the following formulas (6a) to (6c).
  • M represents a divalent metal ion belonging to Groups 8 to 12 of the periodic table
  • X represents a monovalent anionic monodentate ligand
  • L represents The tridentate ligand represented by the formula (1) is represented
  • solv represents a guest molecule such as a solvent molecule used in the synthesis
  • SA represents a skeleton-forming aromatic compound
  • a, b and c represent arbitrary natural numbers.
  • the polynuclear metal complex using the TPT represented by the formula (4a) as L is a form in which a guest molecule such as a solvent has been incorporated so far.
  • the molecular structure is determined by single crystal X-ray structural analysis, and is particularly suitable as a polynuclear metal complex used in the present invention.
  • polynuclear metal complexes examples include polynuclear metal complexes represented by the following formulas (7a) to (7d).
  • PhNO 2 Nitrobenzene TPH: Triphenylene PER: Perylene MeOH: Methanol DCB: 1,2-dichlorobenzene
  • the three-dimensional skeleton of the polynuclear metal complex 1 is shown in FIGS. 2 (a) to 2 (d).
  • the three-dimensional skeleton of the polynuclear metal complex 1 is composed of two molecular chains 1a and 1b.
  • each zinc (II) ion is coordinated in a tetracoordinate tetrahedral form by two TPT pyridyl groups and two iodide ions.
  • the structures containing the zinc (II) ions are three-dimensionally connected by TPT to form respective molecular chains [FIG. 2 (a)].
  • Each of the molecular chains 1a and 1b has a closed cyclic chain structure composed of a TPT10 molecule and a Zn10 atom as the shortest closed cyclic chain structure [FIG. 2 (b)].
  • These molecular chains 1a and 1b can be regarded as a helical hexagonal three-dimensional network structure having a pitch along the (010) axis of 15 mm [FIG. 2 (c)].
  • the molecular chains 1a and 1b do not share the same zinc (II) ion and are independent of each other. Then, a three-dimensional skeleton organized as a whole is formed by interpenetrating each other in a nested manner so as to share the same space.
  • the single crystal of the polynuclear metal complex 1 having a three-dimensional skeleton has one kind of regularly arranged pores [FIG. 2 (d)].
  • the porosity of the single crystal of the polynuclear metal complex 1 is 50%.
  • the diameter of the inscribed circle of the single crystal pore of the polynuclear metal complex 1 is 5 to 8 mm.
  • Examples of the polynuclear metal complex represented by the formula (7b) include [(ZnBr 2 ) 3 (TPT) 2 (PhNO 2 ) 5 (H 2 O)] n (polynuclear metal complex 2 described in JP-A-2008-214318. And all or part of the reaction solvent molecules in the polynuclear metal complex 2 are replaced with a substitution solvent.
  • the polynuclear metal complex 2 has a skeleton similar to the three-dimensional skeleton of the polynuclear metal complex 1 except that (ZnI 2 ) is replaced by (ZnBr 2 ).
  • the shape and size of the pores of the single crystal of the polynuclear metal complex 2 and the porosity were almost the same as those of the single crystal of the polynuclear metal complex 1.
  • the three-dimensional skeleton of the polynuclear metal complex 3 is shown in FIGS.
  • the three-dimensional skeleton of the polynuclear metal complex 3 is composed of two molecular chains 1A and 1B and a triphenylene molecule that is a skeleton-forming aromatic compound.
  • each zinc (II) ion has two iodide ions and two TPT pyridyl groups coordinated in a tetracoordinate tetrahedral form. Then, the molecular chains are formed by three-dimensionally connecting the structures containing zinc (II) ions with TPT.
  • the molecular chains 1A and 1B do not share the same zinc (II) ion and are independent of each other. Then, a three-dimensional skeleton organized as a whole is formed by interpenetrating each other in a nested manner so as to share the same space.
  • the triphenylene molecule (2) contains tris (4-pyridyl) triazine [TPT (1a)] with a molecular chain 1A and tris (4-pyridyl) triazine [TPT ( 1b)] is firmly inserted (intercalated) with the ⁇ plane [FIG. 3B].
  • the triphenylene molecule is stabilized by the ⁇ - ⁇ interaction between TPT (1a) and TPT (1b) and functions as a part of the three-dimensional skeleton of the polynuclear metal complex 3.
  • FIG.3 (b) is a figure when the part enclosed with the line in Fig.3 (a) is seen from the side.
  • the single crystal of the polynuclear metal complex 3 has two types of regularly arranged pores (pores A and B) [FIG. 3 (c)].
  • the pores A and B are regularly formed between stacked structures in which TPT and TPH are alternately stacked.
  • the pore A has a substantially cylindrical shape and is substantially surrounded by hydrogen atoms present on the side edges of the infinite number of stacked TPTs and TPHs on the ⁇ plane.
  • the pore B has a substantially triangular prism shape, and among the three surfaces forming the triangular prism, two are surrounded by the ⁇ plane of the TPT, and the other is an infinite number of stacked TPT and TPH.
  • the pores A and B have an elongated shape that is slightly meandered.
  • the porosity of the single crystal pores of the polynuclear metal complex 3 is 28%.
  • the diameter of the inscribed circle of the pore A of the single crystal of the polynuclear metal complex 3 is 5 to 8 mm.
  • the diameter of the inscribed circle of the single crystal pore B of the polynuclear metal complex 3 is 5 to 8 mm.
  • the polynuclear metal complex 4 has a skeleton structure similar to that of the polynuclear metal complex 3 except that a perylene molecule is inserted between two TPTs instead of the triphenylene molecule of the polynuclear metal complex 3.
  • the shape and size of the pores of the single crystal of the polynuclear metal complex 4 and the porosity were almost the same as those of the single crystal of the polynuclear metal complex 3.
  • the three-dimensional skeleton of the polynuclear metal complex 5 is shown in FIG.
  • the polynuclear metal complex 5 has a [Co 6 (TPT) 4 ] structure composed of six cobalt ions and four TPTs as structural units.
  • This structural unit has an octahedral three-dimensional shape, and cobalt ions are arranged at six vertices of the octahedron [FIG. 4B].
  • cobalt (II) ion four pyridyl groups of TPT and two thiocyanate ions are coordinated in a hexacoordinate octahedral form.
  • FIG. 4B is an enlarged view of a portion surrounded by a line in FIG.
  • the structural unit has a hollow inside.
  • the porosity of the single crystal of the polynuclear metal complex 5 is 78%. This value is a value calculated by combining the pores and the hollow volume.
  • the diameter of the inscribed circle of the single crystal pores of the polynuclear metal complex 5 is 10 to 18 mm.
  • PCP porous coordination polymer
  • MOF metal organic structure
  • the method for synthesizing the polynuclear metal complex is not particularly limited, and a known method can be used.
  • the Sigma-Aldrich brochure published in September 2012 includes multidentate ligands, etc.
  • Hydrothermal method in which a hydrothermal reaction is carried out by heating; a microwave method in which a solvent, a polydentate ligand, a metal ion, etc. are placed in a container and microwave irradiation; a solvent, a polydentate ligand in the container , Ultrasonic methods of putting metal ions, etc., and irradiating ultrasonic waves; solid-phase synthesis methods of mechanically mixing polydentate ligands, metal ions, etc. without using a solvent; Using this method, a single crystal of a polynuclear metal complex can be obtained.
  • the solution method is preferably used.
  • the solvent solution of the second solvent of the metal ion-containing compound is added to the solvent solution of the first solvent of the polydentate ligand, and is kept at 0 to 70 ° C. for several hours to several days. The method of leaving still is mentioned.
  • the metal ion-containing compound is not particularly limited.
  • a compound represented by the formula: MXn can be mentioned.
  • M represents a metal ion
  • X represents a counter ion
  • n represents the valence of M.
  • X include F—, Cl—, Br—, I—, SCN—, NO 3 —, ClO 4 —, BF 4 —, SbF 4 —, PF 6 —, AsF 6 —, CH 3 CO. 2- and the like.
  • Reaction solvents (first solvent and second solvent) used include aromatic hydrocarbons such as benzene, toluene, xylene, chlorobenzene, 1,2-dichlorobenzene, nitrobenzene; n-pentane, n-hexane, n -Aliphatic hydrocarbons such as heptane; Alicyclic hydrocarbons such as cyclopentane, cyclohexane and cycloheptane; Nitriles such as acetonitrile and benzonitrile; Sulfoxides such as dimethyl sulfoxide (DMSO); N, N-dimethyl Amides such as formamide and n-methylpyrrolidone; ethers such as diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane and 1,4-dioxane; alcohols such as methanol, ethanol and isopropyl alcohol; acetone, methyl ethyl ket
  • the first solvent and the second solvent that are not compatible with each other (that is, separated into two layers).
  • a method of using nitrobenzene, dichlorobenzene, a mixed solvent of nitrobenzene and methanol, a mixed solvent of dichlorobenzene and methanol as the first solvent, and using methanol as the second solvent may be mentioned.
  • the polynuclear metal complexes 1 to 5 can be synthesized according to the methods described in the above documents.
  • a step of bringing a single crystal of a porous compound into contact with a solvent solution containing a metabolite is not particularly limited.
  • a method of immersing the single crystal in a solvent solution containing a metabolite a method of passing the solvent solution containing a metabolite through the capillary after the single crystal is packed in a capillary, and the like.
  • a method in which a single crystal of a porous compound is immersed in a solvent solution containing a metabolite is preferable because the solvent solution containing the metabolite can be efficiently brought into contact with the single crystal of the porous compound.
  • the A value calculated from the following formula (I) is 100 or less, preferably 0.1 to 30, more preferably 1 to 5. It is preferable to immerse an amount of the single crystal of the porous compound.
  • X represents the mass of the metabolite in the solvent solution
  • Y is assumed to fill all pores and hollows in the single crystal of the porous compound with a substance having a specific gravity of 1. It represents the mass of the material having the specific gravity of 1 that is sometimes required.
  • the metabolite molecules are sufficiently taken into the pores and hollows of the single crystal of the porous compound, and it becomes easy to obtain a good quality crystal structure analysis sample.
  • the target crystal structure analysis sample can be obtained, but an effect commensurate with it cannot be obtained, and metabolites are likely to be wasted.
  • the number of single crystals to be immersed is not particularly limited. When the amount of the metabolite is extremely small, a target crystal structure analysis sample can be obtained by immersing one single crystal. When there is a sufficient amount of metabolites, two or more single crystals of the same kind of porous compound may be immersed, or single crystals of different kinds of porous compounds may be immersed simultaneously.
  • the solvent solution may be concentrated by volatilizing the solvent under mild conditions. By performing this treatment, metabolite molecules can be efficiently taken into the pores and hollows of the single crystal of the porous compound.
  • the immersion conditions (concentration conditions) at this time are not particularly limited, but the temperature of the solvent solution is preferably 0 to 180 ° C., more preferably 0 to 80 ° C., and further preferably 20 to 60 ° C.
  • the immersion time (concentration time) is usually 6 hours or more, preferably 12 to 168 hours, more preferably 24 to 78 hours.
  • the volatilization rate of the solvent is preferably 0.1 to 1000 ⁇ L / 24 hours, more preferably 1 to 100 ⁇ L / 24 hours, and further preferably 5 to 50 ⁇ L / 24 hours.
  • the volatilization rate of the solvent is too fast, there is a possibility that a high-quality crystal structure analysis sample cannot be obtained.
  • the volatilization rate of the solvent is too slow, it is not preferable from the viewpoint of working efficiency.
  • the temperature at which the solvent is volatilized is usually 0 to 120 ° C., preferably 15 to 60 ° C., although it depends on the boiling point of the organic solvent used.
  • the operation of immersing the porous compound single crystal in a solvent solution containing a metabolite, volatilizing the solvent, and concentrating the solvent solution can be performed under normal pressure or under reduced pressure. You may carry out under pressure.
  • the pressure during the operation of volatilizing the solvent and concentrating the solvent solution is usually 1 to 1 ⁇ 10 6 Pa, preferably 1 ⁇ 10 to 1 ⁇ 10 6 Pa.
  • the volatilization rate of the solvent can be adjusted appropriately by adjusting the temperature and pressure during the operation of concentrating the solvent solution.
  • the molecules of the solvent used in the solvent solution are incorporated into the pores and hollows of the single crystal of the porous compound.
  • a process may be provided. By providing this process, the solvent molecules used during the synthesis of the porous compound are removed from the pores and hollows and replaced with a solvent that can be easily replaced with metabolite molecules. A sample can be produced efficiently.
  • the single crystal to be used is immersed in the solvent used for preparing the solvent solution containing the metabolite in advance.
  • a method is mentioned.
  • the immersion conditions at this time are not particularly limited, but the temperature of the solvent is usually 0 to 70 ° C., preferably 10 to 60 ° C., more preferably 20 to 50 ° C., and the immersion time is usually 6 hours or more. , Preferably 12 to 168 hours, more preferably 24 to 78 hours.
  • sample for crystal structure analysis obtained by the method of the present invention has metabolite molecules regularly arranged in the pores and hollows of the single crystal of the porous compound.
  • metabolite molecules are regularly arranged” means that the metabolite molecules are not disturbed to such an extent that the structure can be determined by crystal structure analysis. It means that it is regularly accommodated in the hole and hollow.
  • the sample for crystal structure analysis obtained by the method of the present invention was irradiated with MoK ⁇ rays (wavelength: 0.71 ⁇ ) generated at a tube voltage of 24 kV and a tube current of 50 mA, and diffracted X-rays were detected by a CCD detector. Sometimes it is preferable to be able to determine the molecular structure with a resolution of at least 1.5 mm.
  • the metabolite molecules are contained in all the pores and hollows of the single crystal of the porous compound. Need not be included.
  • the solvent used in the solvent solution of the metabolite may be incorporated into a part of the pores and hollows in the single crystal of the porous compound.
  • the crystal structure analysis sample obtained by the method of the present invention preferably has a metabolite molecule occupancy of 10% or more.
  • the occupancy is a value obtained by crystal structure analysis.
  • the amount of the guest molecule (metabolite molecule) in the ideal inclusion state is 100%, the guest molecule actually present in the single crystal is It represents the quantity.
  • the sample for crystal structure analysis obtained by the method of the present invention is a metabolite which is available only in a trace amount or a liquid metabolite at room temperature (20 ° C.), the structure should be clarified by the crystal structure analysis method. Is possible.
  • the target crystal structure analysis sample is obtained by the method of the present invention because the single crystal of the porous compound used in the present invention aligns the orientation of the metabolite molecules as described below. It is thought that it is functioning as.
  • the metabolite molecular structure determination method of the present invention uses the crystal structure analysis sample obtained by the method of the present invention to determine the metabolite molecular structure by the crystal structure analysis method. It is characterized by doing.
  • any of X-ray diffraction and neutron diffraction can be used.
  • a method similar to the conventional one can be used except that the sample for crystal structure analysis obtained by the above method is mounted instead of the conventional single crystal. it can.
  • the molecular structure of the metabolite can be determined more efficiently by using the crystal structure analysis data of the parent compound of the metabolite.
  • the crystal structure analysis data (electron density diagram) of the crystal structure analysis sample obtained using the method for preparing the crystal structure analysis sample of the present invention and the parent compound molecule of the metabolite are: Crystal structure analysis data (electron density diagram) for a crystal structure analysis sample regularly arranged in the pores and / or hollows of the same single crystal (single crystal of the same porous compound) as the single crystal. ) Can be used to know structural changes due to metabolic reactions. By utilizing this knowledge, the molecular structure of the metabolite can be determined more efficiently and more easily.
  • the molecular structure determination method of the present invention even a trace amount of a metabolite can efficiently perform a crystal structure analysis and determine its molecular structure. Moreover, even if it is a liquid metabolite at normal temperature, the molecular structure can be determined by using a crystal structure analysis sample that includes the metabolite. Therefore, according to the molecular structure determination method of the present invention, in the development of drugs, agricultural chemicals, foods, etc., the cost and time required for the analysis of metabolites, which are important in ensuring safety and confirming usefulness, are greatly reduced. can do.
  • Example 1 61 mg of dry baker's yeast and 134 mg of sucrose were added to 1 ml of water to obtain a yeast suspension.
  • 0.1 ml of an adrenosterone ethanol solution 0.1 mg / ml
  • diethyl ether was added to the obtained suspension, and this was stirred for 12 hours and then allowed to stand.
  • the diethyl ether layer was extracted, and the metabolites in the suspension were extracted with diethyl ether.
  • Diethyl ether was added so that the concentration of the metabolite in diethyl ether was about 1 ⁇ g / ⁇ l to obtain a sample solution.
  • FIG. 5 shows the crystal structure obtained
  • FIG. 6 shows the structural formula of the analyzed metabolite.
  • FIG. 7 shows the crystal structure obtained
  • FIG. 8 shows the structural formula of the metabolite analyzed.
  • Example 3 In a microvial, 5 ⁇ g of 6 ⁇ -hydroxytestosterone (Aldrich, which is known as a metabolite of testosterone produced by metabolizing testosterone with the metabolic enzyme CYP450), 5 ⁇ l of methylene chloride and 45 ⁇ l of cyclohexane. The whole volume was made into a uniform solution. The porous compound 1 crystal obtained in Production Example 1 is immersed in the obtained solution, and the solution is concentrated by slowly distilling off the solvent at 50 ° C. over 2 days. Obtained. Table 3 shows the crystallographic data obtained by mounting the obtained sample for crystal structure analysis on an X-ray structure analyzer and analyzing the crystal structure.
  • 6 ⁇ -hydroxytestosterone Aldrich, which is known as a metabolite of testosterone produced by metabolizing testosterone with the metabolic enzyme CYP450
  • FIG. 9 shows the crystal structure obtained
  • FIG. 10 shows the structural formula of the analyzed metabolite.
  • the obtained metabolite was dissolved in a microvial by adding 5 ⁇ l of methylene chloride and 45 ⁇ l of cyclohexane.
  • a crystal of porous compound 1 obtained in Production Example 1 was immersed in the obtained solution, and the solvent was slowly distilled off at 50 ° C. for 5 days to obtain a single crystal sample. This single crystal sample was mounted on an X-ray structure analysis apparatus, and crystal structure analysis was performed. The crystallographic data is shown in Table 4.
  • FIG. 11 shows the crystal structure obtained
  • FIG. 12 shows the structural formula of the metabolite analyzed.
  • FIG. 13 shows the crystal structure obtained
  • FIG. 14 shows the structural formula of the metabolite analyzed.
  • Example 6 DDT was metabolized in the same manner as in the above (Example 1-5), and components with an outflow time of about 9 minutes were separated by analytical liquid chromatography (column INERTSIL NH2, n-hexane 100%). The solvent of the obtained fraction was distilled off, and 5 ⁇ l of methylene chloride and 45 ⁇ l of cyclohexane were added and dissolved in the microvial. A crystal of porous compound 1 obtained in Production Example 1 was immersed in the obtained solution, and the solvent was slowly distilled off at 50 ° C. for 2 days to obtain a single crystal sample. This single crystal sample was mounted on an X-ray structure analysis apparatus, and crystal structure analysis was performed. The crystallographic data is shown in Table 6.
  • FIG. 15 shows the crystal structure obtained
  • FIG. 16 shows the structural formula of the analyzed metabolite.
  • Example 7 Crystals of porous compound 1 obtained in Production Example 1 were added to a microvial, and 45 ⁇ l of cyclohexane and 5 ⁇ l of a solution of 1 mg of adrenosterone in 1 ml of methylene chloride were added. Thereafter, the solvent was slowly distilled off at 50 ° C. over 2 days to obtain a single crystal sample. This single crystal sample was mounted on an X-ray structure analysis apparatus, and crystal structure analysis was performed. When this crystal structure was compared with the crystal structure of the inclusion complex of the metabolite in Example 1, it was found that the positions of the parent compound and the metabolite in the crystal were almost the same.
  • the molecular structure of the metabolite can be determined. It is possible to determine more quickly.
  • Table 7 shows the crystallographic data of the adrenosterone inclusion complex.
  • FIG. 17 shows the crystal structure obtained
  • FIG. 18 shows the structural formula of the metabolite analyzed
  • FIG. 19 shows a comparative diagram of the positions of the parent compound and metabolite in the crystal.

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Abstract

La présente invention concerne un procédé qui est conçu pour préparer un échantillon de cristallographie pour déterminer la structure moléculaire d'un métabolite, et qui est caractérisé en ce qu'il comprend une étape de préparation d'un échantillon de cristallographie est préparé dont les molécules du métabolite sont disposées de façon régulière dans les pores et/ou cavités du monocristal, la préparation étant réalisée au moyen de la mise en contact d'une solution de solvant contenant le métabolite, avec un monocristal d'un composé poreux ayant un squelette 3D, qui est conçu à partir d'une ou plusieurs chaînes moléculaires ou d'une ou plusieurs chaînes moléculaires et d'un composé formant un squelette, et des pores et/ou cavités formés en étant séparés par le squelette 3D et disposés de façon tridimensionnelle régulière.
PCT/JP2014/067233 2013-12-27 2014-06-27 Procédé pour préparer un échantillon de cristallographie et procédé pour déterminer la structure moléculaire de métabolite Ceased WO2015098152A1 (fr)

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CN113287003A (zh) * 2018-11-22 2021-08-20 株式会社理学 单晶x射线结构解析装置用试样保持架组件
EP4029869A1 (fr) * 2021-01-15 2022-07-20 Merck Patent GmbH Complexe métallique polynucléaire cristallin solvaté avec un mélange de solvants non polaires et polaires, ledit complexe métallique polynucléaire cristallin solvaté comprenant un analyte de composé hôte et son utilisation dans un procédé de détermination de la structure moléculaire de l'analyte de composé hôte
EP3885751A4 (fr) * 2018-11-21 2022-10-12 Rigaku Corporation Dispositif et procédé d'analyse structurelle aux rayons x sur monocristal, et unité porte-échantillon associé

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WO2018159692A1 (fr) * 2017-03-01 2018-09-07 国立大学法人 東京大学 Procédé d'identification d'une structure moléculaire

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EP3885751A4 (fr) * 2018-11-21 2022-10-12 Rigaku Corporation Dispositif et procédé d'analyse structurelle aux rayons x sur monocristal, et unité porte-échantillon associé
CN113287003A (zh) * 2018-11-22 2021-08-20 株式会社理学 单晶x射线结构解析装置用试样保持架组件
EP4029869A1 (fr) * 2021-01-15 2022-07-20 Merck Patent GmbH Complexe métallique polynucléaire cristallin solvaté avec un mélange de solvants non polaires et polaires, ledit complexe métallique polynucléaire cristallin solvaté comprenant un analyte de composé hôte et son utilisation dans un procédé de détermination de la structure moléculaire de l'analyte de composé hôte
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