Tool for crystal structure analysis, and method for manufacturing the same, and method for determining molecular structure of target molecule

Abstract

To provide a tool for crystal structure analysis and a method for manufacturing the same, and a method for determining the molecular structure of a target molecule that allow execution of a series of operations in a molecular structure determination method by a crystal sponge method without contact of an experimental instrument or fingers with a crystal structure analysis sample, and contribute to full automation of the series of operations.SOLUTION: A tool for crystal structure analysis 1, 2 respectively have single crystal holders 1a, 2a, liquid for fixing a single crystal 1b, 2b, and one or two or more crystal structure analysis samples 1c, 2c. The single crystal holder has a shape that can be fixed to a goniometer of a crystal structure analyzer. The liquid for fixing a single crystal has lost its fluidity and is present on a surface of each single crystal holder. The crystal structure analysis sample is covered by the liquid for fixing a single crystal and directly or indirectly fixed to the surface of the single crystal holder. The crystal structure analysis sample regularly accommodates target molecules in a single crystal having a plurality of regularly arranged target molecule accommodation spaces.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to an instrument for crystal structure analysis, a method for producing the same, and a method for determining the molecular structure of a target molecule. [Background technology]

[0002] Conventionally, single crystal structure analysis methods such as single crystal X-ray diffraction have been known as one of the highly reliable methods for determining molecular structures. However, since single crystal structure analysis requires the use of high-quality single crystals as measurement samples, when performing crystal structure analysis of unknown compounds or compounds that are difficult to crystallize, it can take a lot of time and effort to prepare the measurement samples.

[0003] As a method for solving this problem, a method using a single crystal of a porous polymer compound is known. For example, Patent Document 1 describes a method in which molecules of a compound to be analyzed are regularly arranged within the pores of a single crystal of a polymer complex, and the resulting guest molecule inclusion complex is used as a measurement sample to perform crystal structure analysis (the so-called "method for determining molecular structure by the crystalline sponge method"). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2014 / 038220 Summary of the Invention [Problem to be solved by the invention]

[0005] Figure 1 is a simple diagram showing an example of the procedure for determining the molecular structure of a molecule of a target compound (hereinafter, the "molecule of the target compound" may be referred to as the "target molecule") using the crystalline sponge method. In this example, first, as shown in Figs. 1(a) and (b), a raw material solution of the polymer complex is prepared, and then allowed to stand to precipitate single crystals of the polymer complex. Next, as shown in Figures 1(c) and (d), a portion of the obtained single crystal is sucked up together with the solution using a pipette or the like, and the single crystal is transferred to a container containing the solution in which the target molecule has been dissolved, and the single crystal is immersed in the solution in which the target molecule has been dissolved. Next, as shown in FIG. 1(e), the solution in which the target molecules are dissolved is concentrated while the target molecules are incorporated into the single crystal. Next, as shown in Figure 1(f) and (g), the single crystal incorporating the target molecule is fixed to the tip of a glass rod using adhesive or the like, or is packed into a capillary and fixed, or is fixed to a sample loop, and then the single crystal is placed in a crystal structure analyzer for measurement.

[0006] As described above, conventional methods require tasks in which laboratory equipment or fingers come into direct contact with the single crystal, such as moving the single crystal with a pipette or fixing the single crystal before performing crystal structure analysis, and these tasks pose the risk of deteriorating the single crystal. Furthermore, in order to perform structural analysis more efficiently, it would be desirable to fully automate the series of steps in the molecular structure determination method using the crystalline sponge method; however, single crystals can be very fragile, and it has been difficult to mechanize tasks in which laboratory equipment, etc., as described above, directly contacts the single crystals.

[0007] The present invention aims to solve these problems, and to provide a crystal structure analysis instrument and a manufacturing method thereof, and a method for determining the molecular structure of a target molecule, which enable a series of steps in a molecular structure determination method using the crystalline sponge method to be performed without bringing laboratory equipment or fingers into contact with the crystal structure analysis sample (and its precursor, the single crystal), and which contributes to the complete automation of the series of steps. [Means for solving the problem]

[0008] In order to solve the above problems, the present inventors have extensively investigated a series of steps in a molecular structure determination method using the crystalline sponge method. As a result, we discovered that by using a single crystal holder having a shape that can be fixed to the goniometer of a crystal structure analysis device and a single crystal fixing liquid that functions as a fixative or fixing assistant for crystal structure analysis samples (and their precursor single crystals), it is possible to efficiently obtain a crystal structure analysis instrument having the desired characteristics, and thus completed the present invention.

[0009] Thus, according to the present invention, there are provided the following crystal structure analysis instruments (1) to (10), methods for manufacturing the crystal structure analysis instruments (11) to (13), and methods for determining the molecular structure of a target molecule (14) and (15).

[0010] [1] A crystal structure analysis apparatus having a single crystal holder, a single crystal fixing liquid, and one or more crystal structure analysis samples, wherein the single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analysis device directly or via other parts, the single crystal fixing liquid is present on the surface of the single crystal holder in a state in which it has lost fluidity, the crystal structure analysis sample is fixed directly or indirectly to the surface of the single crystal holder while being covered with the single crystal fixing liquid, and the crystal structure analysis sample is an apparatus for crystal structure analysis comprising a single crystal having a plurality of regularly arranged target molecule storage spaces, and the target molecules are regularly stored in the target molecule storage spaces. [2] The crystal structure analysis device according to [1], wherein the single crystal holder has a shape of a multi-hole plate, a dish, a plate or a thin film in whole or in part. [3] The crystal structure analysis apparatus according to [1] or [2], wherein the single crystal holder is made entirely or in part of glass, resin or carbon. [4] The crystal structure analysis instrument according to any one of [1] to [3], wherein the liquid for fixing the single crystal on the surface of the single crystal holder is a droplet having a volume of 0.01 to 500 μL. [5] The crystal structure analysis instrument according to [4], wherein the number of droplets of the single crystal fixing liquid on the surface of the single crystal holder is 1 to 500. [6] The crystal structure analysis instrument according to [4] or [5], wherein the number of the crystal structure analysis samples contained in one droplet of the single crystal fixing liquid is 1 to 1000. [7] The crystal structure analysis instrument according to any one of [1] to [6], wherein the crystal structure analysis sample has a longest side length of 500 μm or less when the smallest rectangular parallelepiped capable of containing the crystal structure analysis sample is imagined. [8] The crystal structure analysis instrument according to any one of [1] to [7], wherein the single crystal is grown on the surface of the single crystal holder. [9] The crystal structure analysis instrument according to any one of [1] to [8], wherein the single crystal is a crystal of a polynuclear metal complex.

[10] The crystal structure analysis apparatus according to any one of [1] to [9], wherein the single crystal holder is a resin multi-hole plate, and the crystal structure analysis sample has a length of the longest side of the smallest imaginary rectangular parallelepiped capable of containing the crystal structure analysis sample, which is 100 μm or less.

[11] A method for manufacturing a crystal structure analysis instrument according to any one of [1] to

[10] above, comprising the steps of: bringing a single crystal preparation solution into contact with a single crystal holder to precipitate a single crystal on the surface of the single crystal holder (step a); bringing a single crystal fixing liquid into contact with the single crystal precipitated in step a (step b); and bringing target molecules into contact with the single crystal on the single crystal holder to regularly accommodate the target molecules in the target molecule accommodation space of the single crystal (step c).

[12] A method for manufacturing a crystal structure analysis instrument according to

[11] , characterized in that steps b and c are carried out simultaneously using a liquid containing a single crystal fixing liquid and a target molecule.

[13] A method for manufacturing a crystal structure analysis instrument according to

[11] or

[12] , characterized in that steps a, b, and c are carried out without laboratory instruments or fingers coming into contact with either the single crystal or the crystal structure analysis sample.

[14] A method for determining the molecular structure of a target molecule by crystal structure analysis using the crystal structure analysis instrument according to any one of [1] to

[10] above, characterized in that the crystal structure analysis sample constituting the crystal structure analysis instrument is placed in a crystal structure analyzer without contacting the crystal structure analysis sample with laboratory instruments or fingers.

[15] A method for determining the molecular structure of a target molecule by crystal structure analysis, comprising the following steps 1 and 2: [Step 1] A step of irradiating a crystal structure analysis sample constituting the crystal structure analysis instrument according to the above

[10] with synchrotron X-rays and collecting diffraction intensity data. [Step 2] A step of performing the above step 1 for a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data. Effect of the Invention

[0011] According to the present invention, a series of steps in a molecular structure determination method using the crystalline sponge method can be performed without bringing laboratory equipment or fingers into contact with the crystal structure analysis sample (and its precursor, the single crystal), and the present invention provides a crystal structure analysis instrument and a manufacturing method thereof, and a method for determining the molecular structure of a target molecule, which contribute to the complete automation of the series of steps. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 is a simplified diagram of a general method for determining molecular structure by the crystalline sponge method. [Diagram 2] FIG. 1 is a schematic diagram showing an example of a crystal structure analysis instrument according to the present invention. [Diagram 3] FIG. 1 is a diagram showing a simplified manufacturing process of a crystal structure analysis device having a multi-hole plate-shaped single crystal holder. [Figure 4] 1 is a diagram simply illustrating a manufacturing process of a crystal structure analysis tool having a plate-shaped single crystal holder. [Diagram 5] FIG. 2 is a diagram showing the results of crystal structure analysis carried out in Production Example 1. [Figure 6] FIG. 2 is a diagram showing the results of crystal structure analysis carried out in Reference Example 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present invention will be described in detail below, divided into the following sections: 1) an instrument for crystal structure analysis and a method for producing the same, and 2) a method for determining the molecular structure of a target molecule.

[0014] 1) Crystal structure analysis equipment and its manufacturing method The crystal structure analysis instrument of the present invention is an instrument for crystal structure analysis having a single crystal holder, a single crystal fixing liquid, and one or more crystal structure analysis specimens, the single crystal holder having a shape that allows it to be fixed to a goniometer of a crystal structure analysis apparatus directly or via another part. The single crystal fixing liquid is present on the surface of the single crystal holder in a state in which it has lost fluidity, the crystal structure analysis specimen is fixed directly or indirectly to the surface of the single crystal holder while being covered with the single crystal fixing liquid, and the crystal structure analysis specimen is composed of a single crystal having a plurality of regularly arranged target molecule accommodation spaces, and the target molecules are regularly accommodated in the target molecule accommodation spaces.

[0015] As used herein, the term "target molecule" refers to a molecule of a compound to be analyzed. The term "target molecule-accommodating space" refers to the space inside the single crystal in which the target molecule is accommodated. The shape of the space is not limited as long as it can accommodate the target molecule, and may be elongated, for example, like a "pore." The term "orderly arranged" in "plurality of target molecule-accommodating spaces regularly arranged" means that the structure of the molecules surrounding the target molecule-accommodating spaces can be determined by crystal structure analysis. By "the single crystal accommodates the target molecule in an orderly manner" is meant that the single crystal accommodates the target molecule in such a way that the molecular structure of the target molecule can be determined by crystal structure analysis. In this specification, the term "crystal structure analysis sample" refers to a single crystal in which a target molecule is included in the target molecule accommodation space. In addition, in this specification, the term "single crystal" refers, in principle, to a crystal having single crystallinity and not having a target molecule in the target molecule accommodation space (i.e., a precursor of a crystal structure analysis sample). However, in some cases, the term "single crystal" is used to include a crystal having a target molecule in the target molecule accommodation space (i.e., a crystal structure analysis sample).

[0016] The crystal structure solving instrument of the present invention is used in determining the molecular structure of a target molecule by crystal structure analysis. In the crystal structure analysis device of the present invention, the crystal structure analysis sample or its precursor, a single crystal, is fixed to the surface of the single crystal holder. Therefore, in a series of operations in the molecular structure determination method using the crystal sponge method, the single crystal or the crystal structure analysis sample can be moved by holding the single crystal holder without touching the single crystal or the crystal structure analysis sample with laboratory equipment or fingers.

[0017] [Single crystal holder] The single crystal holder constituting the crystal structure analysis instrument of the present invention functions as a member for fixing a crystal structure analysis sample or a single crystal which is a precursor thereof.

[0018] The single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analyzer directly or via another part. Because the single crystal holder has such a shape, it is possible to omit the task of fixing the crystal structure analysis sample (a single crystal incorporating a target molecule) to the tip of a glass rod or inside a capillary, which has traditionally been performed prior to crystal structure analysis.

[0019] The shape of the single crystal holder that can be fixed to the goniometer of the crystal structure analyzer includes, for example, a capillary shape, a rod shape, a plate shape, etc. However, the shape is not limited to these. When the single crystal holder has a capillary or rod-shaped shape that can be fixed to the goniometer of a crystal structure analysis apparatus, the single crystal holder can be efficiently fixed to the goniometer of a crystal structure analysis apparatus by utilizing the goniometer head of a conventional crystal structure analysis apparatus.

[0020] From the viewpoint of efficiently fixing a crystal structure analysis sample to the surface of a single crystal holder using a single crystal fixing liquid, and from the viewpoint of irradiating the crystal structure analysis sample with X-rays or the like to obtain sufficient diffraction intensity data, it is preferable that the shape of the entire or part of the single crystal holder is multi-hole plate-like, dish-like, plate-like, or thin film-like. In other words, since at least the portion of the single crystal holder where the crystal structure analysis sample is fixed has the above-mentioned shape, the crystal structure analysis sample is efficiently fixed by the single crystal fixing liquid, and sufficient diffraction intensity data can be easily obtained.

[0021] The single crystal holder is preferably made entirely or partially of glass, resin or carbon. In general, in single crystal structure analysis using X-rays or electron beams, a single crystal is fixed on a substrate made of glass, resin or carbon and then measured. Therefore, by making at least a portion of the single crystal holder from glass, resin or carbon, the crystal structure analysis sample fixed to that portion can be suitably used as a measurement sample for single crystal structure analysis using X-rays or electron beams.

[0022] The single crystal holder may be subjected to a surface treatment, such as a chemical treatment with an oxidizing agent, an acid treatment, an alkali treatment, a primer treatment, or a silane compound treatment, or a physical treatment such as a corona discharge treatment or a plasma treatment. In the crystal structure analysis apparatus of the present invention, the crystal structure analysis sample is fixed to the surface of the single crystal holder while being covered with a single crystal fixing liquid. However, as described below, this crystal structure analysis sample may or may not be fixed to the surface of the single crystal holder in the absence of a single crystal fixing liquid. In the single crystal holder that has been subjected to the above-mentioned surface treatment, a crystal structure analysis sample tends to be more reliably fixed on its surface even in the absence of a single crystal fixing liquid.

[0023] [Liquid for fixing single crystals] The single crystal fixing liquid constituting the crystal structure analysis instrument of the present invention functions as a fixing agent or fixing assistant for the crystal structure analysis sample. In this specification, a fixing agent for a crystal structure analysis sample refers to a liquid agent used to fix a crystal structure analysis sample that is not fixed to the surface of a single crystal holder to the surface of a single crystal holder, and a fixing aid for a crystal structure analysis sample refers to a liquid agent used to more firmly fix a crystal structure analysis sample that has already been fixed to the surface of a single crystal holder.

[0024] The single crystal fixing liquid is present on the surface of the single crystal holder in a state where it has lost its fluidity. When the single crystal fixing liquid is present on the surface of the single crystal holder in a state in which it has lost its fluidity, the single crystal fixing liquid can function as a fixing agent or fixing assistant for the crystal structure analysis sample. The state in which the single crystal fixing liquid has lost its fluidity refers to a state in which the shape of the single crystal fixing liquid does not change to the extent that crystal structure analysis can be performed using a crystal structure analysis sample covered with the single crystal fixing liquid, even when the crystal structure analysis equipment is tilted.

[0025] The viscosity of the single crystal fixing liquid at 20° C. is preferably 0.1 mPa·s or more, and more preferably 0.2 mPa·s or more. By setting the viscosity of the single crystal fixing liquid to 0.1 mPa·s or more, the single crystal fixing liquid can easily maintain a state in which it has lost fluidity. Furthermore, as long as a desired amount of the single crystal fixing liquid can be discharged to a desired position, there is no particular upper limit on the viscosity of the single crystal fixing liquid.

[0026] The components of the liquid for fixing the single crystal include a solvent, a polymer component, other additives, etc. These may be used in combination. In addition, when only a small amount of target molecule can be obtained, it is preferable to prepare a crystal structure analysis sample after confirming that the components of the single crystal fixing liquid do not pose a risk of dissolving the single crystal or promoting its decomposition.

[0027] Examples of the solvent include aromatic hydrocarbons such as benzene, toluene, xylene, chlorobenzene, 1,2-dichlorobenzene, and nitrobenzene; aliphatic hydrocarbons such as n-butane, n-pentane, n-hexane, and n-heptane; alicyclic hydrocarbons such as cyclopentane, cyclohexane, and cycloheptane; nitriles such as acetonitrile and benzonitrile; sulfoxides such as dimethyl sulfoxide (DMSO); amides such as N,N-dimethylformamide and n-methylpyrrolidone; diethyl ether and tetrahydrofuran. ethers such as 1,2-dimethoxyethane, and 1,4-dioxane; alcohols such as methanol, ethanol, isopropyl alcohol, and t-butyl alcohol; ketones such as acetone, methyl ethyl ketone, and cyclohexanone; cellosolves such as ethyl cellosolve; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, and 1,2-dichloroethane; nitro compounds such as nitromethane and nitroethane; esters such as methyl acetate, ethyl acetate, ethyl lactate, and ethyl propionate; water; and the like.

[0028] Examples of the polymer component include paraffin oil, paratone oil, silicone oil, and polyethylene glycol. Other additives include glycerol, ethylene glycol, methylpentanediol, and the like. Furthermore, commercially available oil-based compositions such as cryoprotectants and immersion oils for objective lenses can also be used as the liquid for fixing the single crystal. In addition, in cases where various additives contained in the single crystal fixing liquid are accommodated in the target molecule accommodation space of the single crystal and affect the analysis results, it is preferable that the liquid does not contain these components.

[0029] The single crystal fixing liquid on the surface of the single crystal holder is preferably a droplet having a volume of 0.01 to 500 μl. When the droplet volume is 0.01 μl or more, a sufficient number of crystal structure analysis samples can be contained in the droplet. Furthermore, when the droplet volume is 500 μl or less, the occurrence of fluidity in the single crystal fixing liquid can be avoided. For the same reason, the droplet volume is preferably 0.1 to 400 μl, more preferably 1 to 300 μl, and even more preferably 10 to 200 μl. When preparing droplets with a large volume, it may be possible to avoid fluidity in the single crystal fixing liquid by selecting a single crystal fixing liquid with high viscosity or high surface tension.

[0030] The number of droplets of the single crystal fixing liquid on the surface of the single crystal holder is not particularly limited, but is usually 1 to 500 droplets, preferably 1 to 400 droplets, and more preferably 1 to 200 droplets.

[0031] The number of crystal structure analysis samples contained in one droplet of the single crystal fixing liquid is not particularly limited, but is usually 1 to 1,000, preferably 1 to 500, more preferably 1 to 200, and even more preferably 1 to 200.

[0032] [Crystal structure analysis sample] The crystal structure analysis sample constituting the crystal structure analysis instrument of the present invention is composed of a single crystal having a plurality of regularly arranged target molecule storage spaces, with target molecules regularly stored in the target molecule storage spaces. In the crystal structure analysis instrument of the present invention, a crystal structure analysis sample is an object to be irradiated with X-rays or the like.

[0033] The crystal structure analysis sample is directly or indirectly fixed to the surface of a single crystal holder while being covered with the single crystal fixing liquid. In this specification, the phrase "the crystal structure analysis sample is directly fixed" refers to a state in which the crystal structure analysis sample is fixed to the surface of the single crystal holder even in the absence of a single crystal fixing liquid. An example of a crystal structure analysis sample fixed directly to the surface of a single crystal holder is a single crystal that precipitates in the presence of a single crystal holder, grows without peeling off from the surface of the single crystal holder, and regularly accommodates target molecules in its target molecule accommodation space, as described below. Furthermore, "the crystal structure analysis sample is indirectly fixed" refers to a state in which the crystal structure analysis sample is fixed to the surface of the single crystal holder due to the presence of a single crystal fixing liquid.

[0034] The size of the crystal structure analysis sample is not particularly limited. When the smallest rectangular parallelepiped capable of accommodating the crystal structure analysis sample is imagined, the length of its long side (longest side) is usually 500 μm or less. However, it is preferable that the crystal structure analysis sample has a size suitable for the crystal structure analysis device to be used. For example, when performing crystal structure analysis using a commercially available X-ray crystal structure analyzer, when imagining the smallest rectangular parallelepiped capable of accommodating a crystal structure analysis sample, the length of its long side is usually 10 to 500 μm, preferably 50 to 500 μm, and the length of its short side (shortest side) is usually 10 to 500 μm, preferably 10 to 200 μm.

[0035] When a plurality of crystal structure analysis samples are irradiated with synchrotron X-rays and the molecular structure of the target compound is determined based on the collected diffraction intensity data, when the smallest rectangular parallelepiped capable of accommodating the crystal structure analysis samples is imagined, the length of its long side is usually 0.1 to 100 μm, preferably 1 to 100 μm, and the length of its short side is usually 0.1 to 100 μm, preferably 1 to 10 μm.

[0036] When performing crystal structure analysis by micro electron diffraction, assuming the smallest rectangular parallelepiped capable of accommodating a crystal structure analysis sample, the length of its long side is usually 0.1 to 1000 nm, preferably 1 to 1000 nm, and the length of its short side is usually 0.1 to 100 nm, preferably 0.1 to 100 nm.

[0037] A single crystal, which is a precursor of a crystal structure analysis sample, has a plurality of regularly arranged target molecule accommodation spaces. A crystal structure analysis sample can be obtained by using a single crystal having a plurality of regularly arranged target molecule accommodation spaces. In other words, since a single crystal has a large number of homogeneous target molecule accommodation spaces, each of the target molecules taken into the single crystal is accommodated in a more stable state, and as a result, the target molecules are accommodated in an orderly manner as a whole.

[0038] The size of the target molecule is not particularly limited as long as the target molecule can enter the target molecule accommodation space of the single crystal. The molecular weight of the target molecule is usually 20 to 3,000, preferably 100 to 2,000.

[0039] The occupancy rate of the target molecule in the crystal structure analysis sample is preferably 20% or more, more preferably 50 to 100%, and even more preferably 80 to 100%. The occupancy rate is a value obtained by crystal structure analysis, and represents the amount of the target molecule actually present in the crystal structure analysis sample, when the amount of the target molecule in an ideal inclusion state is taken as 100%.

[0040] The single crystal is preferably grown on the surface of the single crystal holder. When a single crystal is grown on the surface of the single crystal holder, it becomes easier to obtain a single crystal or a crystal structure analysis sample that is directly fixed to the surface of the single crystal holder, and it becomes easier to obtain a crystal structure analysis instrument with superior durability. Furthermore, even if the single crystal grows in a state where it is peeled off from the single crystal holder, if the single crystal has grown on the surface of the single crystal holder and is present on the surface of the single crystal holder, the single crystal can be indirectly fixed to the surface of the single crystal holder by contacting the single crystal with a single crystal fixing liquid on the single crystal holder. In particular, since there is no need to move the single crystal with a pipette or the like when performing this operation, deterioration of the single crystal can be avoided.

[0041] Since the molecular design of single crystals is relatively easy, crystals of polynuclear metal complexes are preferred. Polynuclear metal complexes are metal complexes that contain two or more metal ions and ligands. The polynuclear metal complex used in the present invention is, for example, a metal complex having a three-dimensional structure containing two or more metal ions and a ligand having two or more coordinating sites, and having an internal space.

[0042] The metal ion is not particularly limited as long as it can form a polynuclear metal complex. The metal ion is preferably an ion of an element selected from the group consisting of Ti, Fe, Co, Ni, Cu, Zn, Ru, Rh, Pd, Cd, Os, Ir, and Pt, and more preferably an ion of an element of Group 8, 9, or 10 of the periodic table. The valence of the metal ion is not particularly limited, and is usually 1 to 4, preferably 1 to 3, and more preferably 2.

[0043] The ligands include those that form the framework or walls of the space that contains the target molecule (hereinafter, these may be referred to as "ligand (α)"), and those that have other roles (such as adjusting the charge of the polynuclear metal complex or occupying vacant coordination sites of the metal ion (hereinafter, these may be referred to as "ligand (β)").

[0044] The ligand (α) is preferably a multidentate ligand having a π-conjugated system. When the ligand (α) is a multidentate ligand having a π-conjugated system, a stable target molecule storage space is easily formed. The multidentate ligand having a π-conjugated system is preferably a multidentate ligand containing an aromatic group as a central skeleton. A multidentate ligand containing an aromatic group as a central skeleton is relatively rigid and has excellent planarity, so that a stable target molecule storage space is easily formed. An example of the ligand (α) is one represented by the following formula (1).

[0045] [ka]

[0046] In formula (1), A is an m-valent group having aromaticity. X is a divalent organic group or a single bond directly connecting A and Y. Y is a coordinating atom or a monovalent group containing a coordinating atom. m represents an integer of 2 to 6. Multiple Xs may be different from each other, and multiple Ys may be different from each other.

[0047] The group represented by A typically has 6-100, preferably 6-60, and more preferably 6-30, atoms (excluding hydrogen atoms). Examples of the group represented by A include a six-membered aromatic group, a group in which a plurality of six-membered aromatic groups are linked by single bonds, and a group having a porphyrin skeleton.

[0048] Examples of the six-membered aromatic group include groups having an aromatic ring such as a benzene ring, a triazine ring, a pyridine ring, and a pyrazine ring. The six-membered aromatic group may have a substituent other than -(-XY). Examples of the substituent include an alkyl group having 1 to 10 carbon atoms; and a halogen atom such as a fluorine atom, a bromine atom, or a chlorine atom.

[0049] Examples of the group represented by A include, but are not limited to, the following: In addition, "*" represents a bond (position of bonding to X).

[0050] [ka]

[0051] [ka]

[0052] In the above formula, M represents a metal ion. Examples of the metal ion include the same as those exemplified as the metal ions constituting the polynuclear metal complex. Among these, zinc ions are preferred.

[0053] The group represented by X typically has 1-30, preferably 2-20, and more preferably 2-10, atoms (excluding hydrogen atoms). Examples of the group represented by X include, but are not limited to, the divalent group represented by A, hydrocarbon groups such as methylene, ethylene, 1,2-ethenediyl, 1,2-ethynediyl (acetylene), p-phenylene, and m-phenylene, amide groups (-C(=O)-NH-), ester groups (-C(=O)-O-), oxymethylene groups (-O-CH2-), and oxyethylene groups (-O-CH2CH2-). The group represented by X may be a group formed by bonding two or more of these groups. Examples of such groups include, but are not limited to, the following:

[0054] [ka]

[0055] The coordinating atom represented by Y includes an oxygen atom, a sulfur atom, a nitrogen atom, a phosphorus atom, and the like. The monovalent group represented by Y typically has 1-20, preferably 1-15, and more preferably 1-10, atoms (excluding hydrogen atoms). Examples of the monovalent group represented by Y include a pyridyl group, an amino group, a hydroxyl group, a deprotonated amide group, a carboxylate group, a sulfonate group, a phosphonate group, a dithiocarboxylate group, a cyano group, and groups containing these groups as a substituent. Examples of the monovalent group represented by Y include the following.

[0056] [ka]

[0057] Examples of the ligand (α) include, but are not limited to, the following:

[0058] [ka]

[0059] [ka]

[0060] [ka]

[0061] As the ligand (α), a ligand other than that represented by formula (1) can also be used. Examples of such a ligand include the following.

[0062] [ka]

[0063] The ligand (β) is preferably a ligand having a relatively low molecular weight, since a low molecular weight ligand is less likely to have adverse effects such as steric hindrance on the coordination of the ligand (α). The ligand (β) may be a monodentate ligand or a chelating ligand.

[0064] The monodentate ligand used as the ligand (β) is an oxide ion (O 2- ) and other divalent anions; hydroxide ion (OH - ), chloride ion (Cl - ), bromide ion (Br - ), iodide ion (I - ), thiocyanate ion (SCN - ) and other monovalent anions; water, ammonia, electrically neutral coordinating compounds such as monoalkylamines, dialkylamines, and trialkylamines; and the like.

[0065] Examples of chelating ligands used as ligand (β) include bidentate chelating ligands such as ethylenediamine, N,N'-dimethylethylenediamine, N,N,N',N'-tetramethylethylenediamine, 2,2'-bipyridyl, and 1,2-cyclohexanediamine, but are not limited to these.

[0066] The single crystal of the polynuclear metal complex may contain components other than metal ions and ligands. Components other than metal ions and ligands contained in the single crystal of a polynuclear metal complex include, for example, molecules capable of affinity interactions with the ligands, such as π-π interactions, π-σ interactions, etc. Single crystals of polynuclear metal complexes containing such molecules tend to have more stable target molecule accommodation spaces.

[0067] Examples of molecules capable of affinity interactions such as π-π interaction and π-σ interaction with a ligand include compounds represented by the following formulas.

[0068] [ka]

[0069] [ka]

[0070] Examples of the polynuclear metal complexes include those represented by the following formulas (2) to (4).

[0071] [ka]

[0072] In formulas (2) to (4), L 1 represents a ligand represented by the formula (1) in which m is 3 (tridentate ligand), and M 1represents a divalent metal ion of Groups 8 to 12 of the periodic table, Q represents a monovalent anionic monodentate ligand, "solv" represents a solvent molecule or the like used during synthesis, and T represents a molecule capable of affinity interaction with the ligand, such as π-π interaction or π-σ interaction. a, b, and n are arbitrary numbers.

[0073] Specific examples of the polynuclear metal complexes represented by formulas (2) to (4) include those represented by the following formulas.

[0074] [ka]

[0075] In the formulas (2a) to (4a), "tpt" represents 2,4,6-tris-(4-pyridyl)-1,3,5-triazine, and "solv", "T", a, b, and n have the same meanings as above.

[0076] Details of these polynuclear metal complexes are described in documents such as JP-A-2006-188560, JP-A-2008-214584, and WO2011 / 062260.

[0077] Further, examples of single crystals of polynuclear metal complexes other than those mentioned above include single crystals formed by three-dimensionally regularly assembling host molecules having internal spaces. Examples of host molecules constituting such single crystals include those shown in the diagram below.

[0078] [ka]

[0079] In the above diagram, Pd represents a palladium ion coordinated with a bidentate chelating ligand (eg, ethylenediamine).

[0080] [ka]

[0081] In the above diagram, Pd represents a palladium ion coordinated with a bidentate chelating ligand (eg, ethylenediamine).

[0082] Details of these host molecules are described, for example, in WO 2018 / 159692.

[0083] [Instrument for crystal structure analysis] 2(a) and (b) are schematic diagrams showing an example of the crystal structure analysis tool of the present invention. FIG. 2(a) is a schematic diagram of a crystal structure analysis device (1) in which a single crystal fixing liquid (1b) that has lost its fluidity is attached to the surface (bottom of the holes) of a multi-hole plate-like single crystal holder (1a), and a crystal structure analysis sample (1c) is incorporated into a droplet of the single crystal fixing liquid (1b). The single crystal holder (1a) has a portion (1d) that can be fixed to a goniometer of a crystal structure analyzer, for example, via fixing members that sandwich at least two faces of a multi-hole plate.

[0084] FIG. 2(b) is a schematic diagram of a crystal structure analysis device (2) in which a single crystal fixing liquid (2b) that has lost its fluidity is attached to the surface of a plate-shaped single crystal holder (2a), and a crystal structure analysis sample (2c) is incorporated into a droplet of the single crystal fixing liquid (2b). The single crystal holder (2a) has a portion (2d) that can be fixed to a goniometer of a crystal structure analyzer, for example, via fixing members that sandwich the plate from the front and back sides.

[0085] The crystal structure analysis tools (1) and (2) have portions to which a crystal structure analysis sample is not fixed, such as the vicinity of the portions (1d) and (2d) of the single crystal holders (1a) and (2a) that can be fixed to a goniometer of a crystal structure analysis apparatus. Therefore, with the crystal structure analysis instrument of the present invention, by gripping the single crystal holder, it is possible to move the crystal structure analysis sample without touching the sample with laboratory instruments or fingers.

[0086] In particular, in the conventional crystalline sponge method, the obtained crystal structure analysis sample (single crystal incorporating the target molecule) was fixed to the tip of a glass rod using adhesive or the like, or was packed into a capillary and fixed, or was fixed to a sample loop, and then the crystal structure analysis sample was placed in a crystal structure analyzer for measurement. This may result in laboratory equipment or fingers coming into contact with the crystal structure analysis sample, which may cause the crystal structure analysis sample to deteriorate.

[0087] On the other hand, the crystal structure analysis instrument of the present invention has a crystal structure analysis sample fixed to the surface of a single crystal holder having a shape that can be fixed to a goniometer of a crystal structure analysis apparatus, either directly or via other parts, so that the crystal structure analysis sample can be placed on the crystal structure analysis apparatus together with the crystal structure analysis instrument. In this manner, according to the crystal structure analysis instrument of the present invention, the crystal structure analysis sample can be placed in the crystal structure analysis apparatus without the need for fixing the crystal structure analysis sample to a glass rod or the like.

[0088] Furthermore, among the crystal structure analysis instruments of the present invention, the single crystal holder is a resin multi-hole plate, and the crystal structure analysis sample is one in which, when the smallest rectangular parallelepiped capable of containing the crystal structure analysis sample is imagined, the longest side has a length of 100 μm or less (hereinafter, this crystal structure analysis instrument may be abbreviated as "crystal structure analysis instrument (A)"), which is suitably used when determining the molecular structure of a target molecule by irradiating synchrotron X-rays.

[0089] That is, when irradiating with synchrotron X-rays, it is usually necessary to prepare multiple fine crystal structure analysis samples, irradiate those crystal structure analysis samples with synchrotron X-rays, and determine the molecular structure of the target molecule based on the collected diffraction intensity data, so it is necessary to reliably irradiate the fine crystal structure analysis samples with synchrotron X-rays. When using a crystal structure analysis instrument (A), the number and positions of crystal structure analysis samples can be known in advance, allowing for more efficient irradiation with synchrotron X-rays.

[0090] Furthermore, with multi-well plates, each fraction obtained by HPLC or other methods can be directly injected into each well, making it possible to clarify the molecular structure of the components contained in each fraction by structural analysis. Furthermore, when a multi-well plate is used, a dispenser used in protein crystallization experiments and the like can be utilized, making it easy to fully automate the series of steps in the molecular structure determination method using the crystalline sponge method.

[0091] The thickness of the bottom of the multi-well plate is usually 1 to 500 μm, preferably 10 to 100 μm. If the thickness of the bottom of the multi-well plate is within the above range, sufficient diffraction intensity data can be obtained.

[0092] The crystal structure analysis sample constituting the crystal structure analysis instrument (A) has a longest side of 100 μm or less, preferably 0.1 to 100 μm, more preferably 1 to 50 μm, when the smallest rectangular parallelepiped capable of accommodating the crystal structure analysis sample is imagined, and the shortest side is usually 0.1 to 100 μm, preferably 1 to 10 μm.

[0093] Each hole in the crystallography tool (A) typically contains a droplet of single crystal mounting liquid. The number of crystal structure analysis samples contained in one droplet is usually 1 to 1000, preferably 5 to 500, and more preferably 10 to 100.

[0094] [Method of manufacturing a crystal structure analysis instrument] The manufacturing method of the present invention for a crystal structure analysis instrument comprises the steps of: contacting a single crystal holder with a single crystal preparation solution to precipitate a single crystal on the surface of the single crystal holder (step a); contacting the single crystal precipitated in step a with a single crystal fixing liquid (step b); and contacting target molecules with the single crystal on the single crystal holder to regularly accommodate the target molecules in the target molecule accommodation space of the single crystal (step c).

[0095] Step a is a step of contacting a single crystal holder with a single crystal preparation solution to precipitate a single crystal on the surface of the single crystal holder.

[0096] A single crystal preparation solution is a solution containing components of a single crystal and from which a single crystal is to be precipitated. For example, in the case of a single crystal of a polynuclear metal complex, a solution containing components of the polynuclear metal complex such as metal ions and ligands is used as the single crystal preparation solution. The solution for preparing a single crystal may be a single-phase system, or may be a two-phase system in which two types of solutions are layered on top of each other.

[0097] Regarding the concentration of the solution for preparing a single crystal, for example, in the case of a single crystal of a polynuclear metal complex, the metal ion concentration is usually 1 μM to 1 M, preferably 1 to 100 mM, and the ligand concentration is usually 1 μM to 1 M, preferably 100 μM to 10 mM. The temperature for precipitating the single crystal is usually from -80 to 150°C, and preferably from 10 to 30°C. The standing time for precipitating the single crystals is usually from 1 minute to 1 month, preferably from 1 hour to 1 week.

[0098] In some cases, it is possible to control the size and quality of the resulting single crystal by adjusting the concentration of the solution for preparing the single crystal and the precipitation conditions.

[0099] When the shape of the portion of the single crystal holder where the single crystal is precipitated (the portion where the crystal structure analysis sample is ultimately fixed) is a multi-hole plate, dish, plate, thin film, etc., step a can be performed by pouring or applying the single crystal preparation solution to that portion and precipitating the single crystal. For example, in Fig. 3 (a simplified diagram showing the manufacturing process of a crystal structure analysis instrument having a multi-hole plate-like single crystal holder), the multi-hole plate-like single crystal holder (1a) in Fig. 3(a) is filled with single crystal preparation solution (3) and left to stand in the state shown in Fig. 3(b), resulting in the precipitation of a single crystal (1e) (Fig. 3(c)). Thereafter, the single crystal preparation solution (3) is removed, and cleaning or other treatment is performed as necessary to obtain a crystal structure analysis sample preparation instrument (1f) (Fig. 3(d)). In this specification, the combination of a single crystal holder and a single crystal precipitated on its surface (but not containing a target molecule) is sometimes referred to as a "crystal structure analysis sample preparation device."

[0100] If the shape of the portion of the single crystal holder where the single crystal is to be precipitated (the portion where the crystal structure analysis sample is ultimately fixed) is plate-like, thin-film-like, etc., step a can be carried out by placing the single crystal preparation solution in a container, immersing the portion of the single crystal holder where the single crystal is to be precipitated in the solution, and precipitating the single crystal. For example, in Fig. 4 (a diagram simply showing the manufacturing process of a crystal structure analysis tool having a plate-shaped single crystal holder), the plate-shaped single crystal holder (2a) in Fig. 4(a) is immersed in a single crystal preparation solution (3) and left to stand in the state of Fig. 4(b), whereby a single crystal (2e) precipitates (Fig. 4(c)). Thereafter, the plate-shaped single crystal holder (2a) is taken out and washed as necessary to obtain a crystal structure analysis sample preparation tool (2f) (Fig. 4(d)).

[0101] The washing treatment can be carried out by bringing a washing solvent into contact with the single crystal or the single crystal holder. The obtained crystal structure analysis sample preparation tool may be subjected to a solvent exchange operation within the single crystal. By carrying out a solvent exchange operation within the single crystal, it may be possible to efficiently accommodate the target molecule. The solvent exchange within the single crystal may be carried out by placing the crystal structure analysis sample preparation tool at rest with the single crystal in contact with the solvent to be exchanged.

[0102] Step b is a step of contacting the single crystal precipitated in step a with a liquid for fixing the single crystal. Step b can be carried out, for example, by discharging a single crystal fixing liquid onto the location where the single crystal is present or onto its periphery.

[0103] Step c is a step of contacting target molecules with the single crystal on the single crystal holder and accommodating the target molecules regularly in the target molecule accommodating spaces of the single crystal. Step c can be carried out, for example, by preparing a solution (or suspension) containing the target molecule and contacting the single crystal with this solution (or suspension), or, if the target molecule is a liquid or gas, by directly contacting the single crystal with the target molecule.

[0104] The solvent of the solution (or suspension) containing the target molecule is appropriately selected from among those that do not dissolve the single crystal and dissolve at least a portion of the target molecule.

[0105] Specific examples of the solvent include the solvents exemplified as components of the single crystal fixing liquid.

[0106] The temperature when step c is carried out is usually −20 to 150° C., preferably 0 to 50° C. The period of step c is usually 1 second to 10 days, preferably 1 second to 5 days, more preferably 1 minute to 1 day.

[0107] Steps b and c are both performed after step a, but there is no particular restriction on the order in which they are performed: step b may be performed before step c, step c may be performed before step b, or step b and step c may be performed simultaneously. It is preferable to carry out steps b and c simultaneously using a liquid containing a single crystal fixing liquid and a target molecule, since this makes it possible to carry out a series of steps in the molecular structure determination method using the crystal sponge method more efficiently.

[0108] In the conventional crystalline sponge method, when a single crystal was moved to bring it into contact with a target molecule, the solution was sucked up using a pipette or other device, making it impossible to avoid contact between the single crystal and the experimental equipment.

[0109] On the other hand, as described below, in the manufacturing method of the crystal structure analysis instrument of the present invention, steps a, b, and c can be performed without laboratory instruments or fingers coming into contact with either the single crystal constituting the instrument for preparing a sample for crystal structure analysis or the crystal structure analysis sample constituting the instrument for crystal structure analysis.

[0110] When the shape of the portion of the single crystal holder on which the single crystal is fixed is a multi-hole plate, a dish, a plate, a thin film, etc., steps b and c can be performed by ejecting or applying a liquid for fixing the single crystal or a solution containing the target molecule, and therefore steps a, b, and c can be performed without moving the single crystal (i.e., without laboratory equipment or fingers coming into contact with the single crystal, etc.). For example, as shown in Figures 3(d) to (g), when manufacturing a crystal structure analysis device having a multi-hole plate-shaped single crystal holder, steps b and c can be carried out simultaneously by pouring a solution (4) containing a single crystal fixing liquid and a target molecule into each hole and leaving it in that state. When the solvent evaporates and the remaining liquid loses its fluidity, a crystal structure analysis device (1) is obtained (Figure 3(g)).

[0111] In addition, when the shape of the part of the crystal holder to which the single crystal is fixed is plate-like, thin film-like, etc., and it is necessary to move the single crystal, it is possible to avoid contact between laboratory equipment or fingers and the single crystal by grasping the part of the single crystal holder where the single crystal is not present. For example, when a crystal structure analysis sample preparation tool (2f) is manufactured using the method of Figures 4(a) to (d), the single crystal can be moved by grasping a portion of the single crystal holder where no single crystal is present, without the single crystal coming into contact with laboratory equipment, fingers, etc. As shown in Figures 4(d) to (g), when manufacturing a crystal structure analysis device having a plate-shaped single crystal holder, a solution (4) containing a liquid for fixing the single crystal and a target molecule is dropped so that the single crystal is covered, and the device is left to stand in this state, whereby operations in steps b and c can be performed. When the solvent evaporates and the remaining liquid loses its fluidity, a crystal structure analysis device (2) is obtained (Figure 4(g)).

[0112] 2) Method for determining the molecular structure of a target molecule The method for determining the molecular structure of a target molecule of the present invention is a method for determining the molecular structure of a target molecule by crystal structure analysis using the crystal structure analysis instrument, and is characterized in that the crystal structure analysis sample constituting the crystal structure analysis instrument is placed in a crystal structure analysis apparatus without laboratory instruments or fingers coming into contact with the crystal structure analysis sample.

[0113] As described above, the crystal structure analysis instrument used in the method for determining the molecular structure of a target molecule of the present invention has a crystal structure analysis sample fixed to the surface of a single crystal holder having a shape that allows it to be fixed to a goniometer of a crystal structure analysis apparatus, either directly or via other parts, so that the crystal structure analysis sample can be placed on the crystal structure analysis apparatus without laboratory equipment or fingers coming into contact with the crystal structure analysis sample that constitutes the crystal structure analysis instrument.

[0114] After placing the crystal structure analysis sample in the crystal structure analyzer, the crystal structure analysis sample is irradiated with X-rays or electron beams in accordance with standard methods, and the molecular structure of the target molecule can be determined based on the obtained diffraction intensity data.

[0115] X-rays are electromagnetic waves with wavelengths of approximately 0.05 to 0.25 nm. In an X-ray tube, thermoelectrons emitted from a filament are accelerated in an electric field and collided with an anode (target: a metal such as Cu or Mo), generating two types of X-rays: continuous X-rays and characteristic X-rays. When thermal electrons collide with a metal anode, K-shell electrons are ejected, creating vacant orbitals. When an outer shell electron transitions to this vacant orbit, X-rays with energy equal to the energy difference between the electron orbitals are emitted. The wavelength of the X-rays is unique to the target atoms, so they are called characteristic X-rays, and these characteristic X-rays are used in crystal structure analysis.

[0116] An electron beam is a stream of electrons moving in a certain direction, and is usually obtained by accelerating thermal electrons in an electric field. While X-rays are a flow of particles (photons) that have no electric charge, electron beams are a flow of particles (electrons) that have a negative charge. Therefore, when an electron beam is irradiated onto a sample, it strongly interacts with the atomic nuclei of the atoms that make up the sample and the electrons around them due to the Coulomb force, so that in crystal structure analysis using an electron beam (microelectron diffraction method), sufficient diffraction intensity data can be obtained even for thin and small crystals that are not suitable for X-ray crystal structure analysis. The electron beam used has an acceleration voltage of preferably 10 to 1000 kV, more preferably 100 to 400 kV.

[0117] In the method for determining the molecular structure of a target molecule of the present invention, when the crystal structure analysis instrument used is the crystal structure analysis instrument (A), the molecular structure of the target molecule can usually be determined by a method having the following steps 1 and 2. [Step 1] A step of irradiating a crystal structure analysis sample constituting a crystal structure analysis instrument (A) with synchrotron radiation X-rays and collecting diffraction intensity data. [Step 2] A step of performing the above step 1 for a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data.

[0118] Step 1 is a step of irradiating a crystal structure analysis sample constituting a crystal structure analysis instrument (A) with synchrotron X-rays and collecting diffraction intensity data. Since the crystal structure analysis sample constituting the crystal structure analysis instrument (A) is very small, when such a crystal structure analysis sample is irradiated with X-rays, it tends to be difficult to obtain sufficient diffraction intensity data. Also, the scattering background due to the materials around the crystal structure analysis sample tends to be relatively high.

[0119] For these reasons, when a general X-ray crystal structure analyzer is used, there is a risk that it may not be possible to determine the molecular structure of the target molecule in the crystal structure analysis sample that constitutes the crystal structure analysis instrument (A); however, by irradiating the crystal structure analysis sample that constitutes the crystal structure analysis instrument (A) with synchrotron X-rays, sufficient diffraction intensity data can be collected and the molecular structure of the target molecule can be determined.

[0120] Crystal structure analysis using synchrotron X-rays is a known technique for determining the molecular structures of biopolymers such as proteins, inorganic compounds, organic compounds, etc. In particular, crystal structure analysis using synchrotron X-rays has the characteristic that sufficient diffraction intensity data can be obtained even if the measurement sample is a microcrystal, and is therefore preferably used when determining the molecular structure of a target molecule in a crystal structure analysis sample constituting the crystal structure analysis instrument (A).

[0121] Synchrotron radiation X-rays are X-rays with high brightness and small beam size, making them suitable for structural analysis of microcrystals. The synchrotron radiation X-rays used have a beam size of 0.1×0.1~100×100μm at the sample position. 2 Preferably, the size is 1×1 to 10×10 μm. 2 is more preferred. The synchrotron radiation X-rays used have a photon flux of 1×10 11 Photons / s or more is preferable, 1×10 12 ~5×10 13 photons / s is preferred.

[0122] Step 2 is a step of performing step 1 on a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data. Step 2 also utilizes known techniques in crystallography using synchrotron X-rays. In other words, in the past, when a sample was irradiated with synchrotron X-rays, the sample would sometimes deteriorate during the irradiation, so molecular structures were determined by irradiating multiple samples with synchrotron X-rays and integrating the data obtained. In recent years, programs used for integrating data have been developed.

[0123] For example, small-wedge data collection is a scheme in which multiple samples are used to collect vibration photographic data at angles of about 5 to 10° per sample. When data processing is performed manually, the data can be selected using BLEND (a program that performs hierarchical clustering based on lattice constants) and then merged.

[0124] In recent years, data collection schemes have also been automated. For example, the SPring-8 beamline BL32XU has been automated as the ZOO system. The ZOO system is an automatic data collection system that combines several component technologies, including KUMA, a technology that collects high-quality data by reducing radiation damage to sample crystals; a high-speed two-dimensional scanning system that uses micro-X-rays to scan and detect crystals that cannot be seen by optical microscope observation; SHIKA, a system that identifies the location of crystals by analyzing the large number of diffraction images output from the scanning system; and KAMO, a technology that almost automatically outputs diffraction data as data to be used for structural analysis.

[0125] When determining the molecular structure of a target molecule in a crystal structure analysis sample that constitutes the crystal structure analysis instrument (A), steps 1 and 2 can be performed more efficiently by using an automatic data collection system such as the ZOO system. EXAMPLES

[0126] The present invention will be described in more detail below with reference to examples, although the present invention is not limited to the following examples in any way.

[0127] [Production Example 1] Production of single crystals A metal ion solution [ZnCl2 (1.5 mM), KOH (0.4 mM)] containing ZnCl2, KOH, and t-butyl alcohol as a solvent, and a ligand solution [tpt (0.44 mM)] containing 2,4,6-tris-(4-pyridyl)-1,3,5-triazine (tpt) and a mixed solvent (nitrobenzene / methanol = 12:1) as a solvent were prepared. 20 μl of the metal ion solution and 100 μl of the ligand solution were mixed to obtain a mixed solution. The resulting mixture was dispensed into each well of a 96-well plate at 1.5 μl using a dispensing device for screening protein crystallization conditions (Gryphon LCP, manufactured by Art Robbins Instruments). After sealing the holes of the 96-well plate with film, the plate was left to stand at 50°C for a day, and crystals with long sides of 20 to 30 μm were formed. The formed crystals were not attached to the bottom of the 96-well plate, but were present inside the droplets (a mixed solvent of t-butyl alcohol, nitrobenzene, and methanol), so their positions did not change even when the 96-well plate was held vertically. For this reason, the following diffraction experiments were performed with the 96-well plate held vertically and fixed to the goniometer of the crystal structure analyzer.

[0128] The beamline was equipped with a Dectris PILATUS3 6M detector at SPring-8 BL45XU, with a wavelength of 0.7750 Å and a 5×5 μm 2 A diffraction experiment was performed by irradiating the obtained single crystal with synchrotron radiation X-rays with a beam size of 100 nm. By integrating 38 pieces of data using KAMO software, structural analysis was performed with sufficient completeness. The molecular structure revealed by crystal structure analysis is shown in Figure 5. Figure 5(a) shows the structure of the asymmetric unit, and Figure 5(b) shows the backbone structure and the target molecule accommodation space. These results showed that the molecular structure was identical to that of known crystalline sponges.

[0129] [Production Example 2] Production of single crystals The same operations as in Production Example 1 were carried out, except that the conditions for leaving the 96-well plate were changed to 40° C. for one day. As a result, crystals with long sides of 15 to 20 μm were formed.

[0130] [Production Example 3] Production of single crystals The same operations as in Production Example 1 were carried out, except that the conditions for leaving the 96-well plate were changed to 25° C. for 2 days. As a result, crystals with a long side length of 15 μm or less were formed.

[0131] Example 1 In the same manner as in Production Example 1, single crystals were grown in each well of a 96-well plate. The 96-well plate was left as it was, and the solvent was evaporated. Then, n-hexane (1 μl) was added to each well, and the 96-well plate was left as it was, and the process of evaporating n-hexane was repeated five times. Then, n-hexane (1 μl) was added to each well, and the single crystal was immersed in n-hexane. A mixture of carvacrol and protectant oil (Cargille, Immersion oil for microscopy Type A) (1 μl) [carvacrol:protectant oil = 95:5 (volume ratio)] was added to each well. The 96-well plate was left as it was, and the n-hexane was evaporated, leaving a highly viscous droplet in each well. The single crystal was captured in the droplet, and did not move even when the 96-well plate was tilted. Therefore, it is believed that by carrying out a diffraction experiment similar to that in Production Example 1, the presence of the guest molecule (carvacrol) can be confirmed.

[0132] [Reference example 1] Using the same method as in Example 1, a single crystal was obtained that was entrapped in a highly viscous liquid droplet. This single crystal was attached to a loop-shaped fixture, and a diffraction experiment was performed using a commercially available crystal structure analyzer (Rigaku Corporation, XtaLAB SynergyCustom). The results of the structural analysis are shown in Figure 6. As shown in Figure 6, it was confirmed that carvacrol was present in the target molecule accommodation space of the crystalline sponge.

[0133] [ka] [Explanation of symbols]

[0134] 1, 2: Instruments for crystal structure analysis 1a, 2a: Single crystal holder 1b, 2b: Liquid for fixing single crystals 1c, 2c: Crystal structure analysis samples 1d, 2d: Sites that can be fixed to the goniometer of a crystal structure analyzer 1e, 2e: Single crystal 1f, 2f: Equipment for preparing samples for crystal structure analysis 3: Solution for single crystal preparation 4: Solution containing single crystal fixing liquid and target molecule

Claims

1. 1. A crystal structure analysis apparatus having a single crystal holder, a single crystal fixing liquid, and one or more crystal structure analysis samples, The single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analyzer directly or via another part, the single crystal fixing liquid is present on the surface of the single crystal holder in a state in which it has lost fluidity; the crystal structure analysis sample is directly or indirectly fixed to a surface of a single crystal holder while being covered with the single crystal fixing liquid; The crystal structure analysis sample is a crystal structure analysis instrument in which a single crystal has a plurality of regularly arranged target molecule accommodating spaces, and target molecules are regularly accommodated in the target molecule accommodating spaces.

2. 2. The crystal structure analysis tool according to claim 1, wherein the single crystal holder is entirely or partially shaped like a multi-hole plate, a dish, a plate or a thin film.

3. 2. The crystal structure analysis tool according to claim 1, wherein the single crystal holder is entirely or partially made of glass, resin or carbon.

4. 2. The crystal structure analysis tool according to claim 1, wherein the liquid for fixing the single crystal on the surface of the single crystal holder is a droplet having a volume of 0.01 to 500 μL.

5. 5. The crystal structure analysis instrument according to claim 4, wherein the number of droplets of the single crystal fixing liquid on the surface of said single crystal holder is 1 to 500.

6. 5. The crystal structure analysis instrument according to claim 4, wherein the number of said crystal structure analysis samples contained in one droplet of said single crystal fixing liquid is 1 to 1000.

7. 2. The crystal structure analysis instrument according to claim 1, wherein the crystal structure analysis sample has a longest side of 500 μm or less when the smallest rectangular parallelepiped capable of containing the crystal structure analysis sample is imagined.

8. 2. The crystal structure analysis tool according to claim 1, wherein the single crystal is grown on a surface of the single crystal holder.

9. 2. The crystal structure analysis instrument according to claim 1, wherein the single crystal is a crystal of a polynuclear metal complex.

10. the single crystal holder is a resin multi-hole plate, 2. The crystal structure analysis instrument according to claim 1, wherein the crystal structure analysis sample has a longest side of 100 μm or less when the smallest rectangular parallelepiped capable of containing the crystal structure analysis sample is imagined.

11. A method for producing the crystal structure analysis instrument according to claim 1, comprising the steps of: A step (step a) of contacting a single crystal holder with a solution for preparing a single crystal and precipitating a single crystal on the surface of the single crystal holder; A step (step b) of contacting the single crystal precipitated in step a with a single crystal fixing liquid; and A method for manufacturing a crystal structure analysis instrument, comprising: a step (c) of contacting target molecules with a single crystal on a single crystal holder, and regularly accommodating the target molecules in the target molecule accommodating space of the single crystal.

12. 12. The method for producing a crystal structure analysis instrument according to claim 11, wherein steps b and c are carried out simultaneously using a liquid containing a single crystal fixing liquid and a target molecule.

13. 12. The method for producing a crystal structure analysis instrument according to claim 11, wherein steps a, b, and c are performed without any laboratory instrument or finger coming into contact with either the single crystal or the crystal structure analysis sample.

14. A method for determining the molecular structure of a target molecule by crystal structure analysis using the crystal structure analysis instrument according to claim 1, comprising: A method for determining the molecular structure of a target molecule, comprising placing a crystal structure analysis sample constituting a crystal structure analysis tool in a crystal structure analysis apparatus without touching the crystal structure analysis sample with laboratory equipment or fingers.

15. A method for determining the molecular structure of a target molecule by crystal structure analysis, comprising the following steps 1 and 2: Step 1 A step of irradiating a crystal structure analysis sample constituting the crystal structure analysis instrument according to claim 10 with synchrotron X-rays and collecting diffraction intensity data. Step 2 A step of performing the step 1 for a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data.

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