Compound having an azobenzene structure, vesicle and method for controlling vesicle structure

Abstract

To provide a compound that has an azobenzene structure useful as a vesicle component.SOLUTION: Provided is a compound that has an azobenzene structure represented by formula (1) (In the formula, one of R1 and one of R2 are -Z, -O-Z, -CO-O-Z, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, etc.; Z is a substituted or unsubstituted aliphatic hydrocarbon group; and the other of R1, the other of R2, and R3 to R10 are hydrogen atom or hydrocarbon group.).SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to a compound having an azobenzene structure, a vesicle, and a method for controlling the structure of a vesicle. [Background technology]

[0002] Liposomes, a type of vesicle, are highly biocompatible and biodegradable and can encapsulate molecules inside, so they have been the subject of much research in the fields of medicine, biotechnology, and other fields.

[0003] For example, when liposomes are used as drug carriers, it is necessary to release drugs by utilizing stimuli responsiveness such as temperature responsiveness and pH responsiveness. For example, it is known that membrane deformation can be induced by changing the fluidity of phospholipid membranes due to temperature changes (see, for example, Non-Patent Document 1).

[0004] In addition, photoresponsive liposomes using an azobenzene compound have been proposed (see, for example, Patent Document 1). Patent Document 1 specifically discloses only an example in which KAON12 is used as the azobenzene compound. In Patent Document 1, the deformation of liposomes is carried out using ultraviolet light, for example, 365 nm. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2011-178689 A [Non-patent literature]

[0006] [Non-Patent Document 1] M. Yanagisawa et al., “Shape deformation of ternary vesicles coupled with phase separation”, Phys. Rev. Lett. 100:148102 (2008). Summary of the Invention [Problem to be solved by the invention]

[0007] As a result of extensive research, the present inventors discovered that vesicles containing a compound having a specific azobenzene structure in the lipid bilayer easily undergo structural changes upon irradiation with light, thereby completing the present invention.

[0008] In other words, the present disclosure aims to provide a method for controlling vesicle structure, in which the structure of the vesicle is changed by irradiation with light, a vesicle that can be used in the control method, and a compound having an azobenzene structure contained in the lipid bilayer of the vesicle. [Means for solving the problem]

[0009] The present inventors have discovered that vesicles containing a specific azobenzene structure in the lipid bilayer can undergo photoisomerization of a compound having an azobenzene structure upon irradiation with light, thereby enabling a structural change of the vesicle, and have completed the present invention.

[0010] An example aspect of this embodiment is described as follows.

[0011] (1) A compound having an azobenzene structure represented by the following general formula (1): [ka] (In the general formula (1), R 1 One of the two and R 2 each independently is -Z, -OZ, -CO-OZ, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, -NH-CO-OZ, -O-CO-NH-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -SZ, -CO-SZ, -S-CO-Z, or -SSZ, Z's each independently represent a substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms, and at least one methylene structure in the substituted or unsubstituted aliphatic hydrocarbon group may be replaced by a structure selected from -O-, -S-, -NH-, -CO-, -SS-, -SO2-, -CO-O-, -CO-NH-, and -NH-CO-O-; R 1 On the other hand, R 2 The other and R 3 ~R 10 are each independently a hydrogen atom or a hydrocarbon group having 1 to 24 carbon atoms. (2) The above R 3 ~R 6 At least two of R 7 ~R 10 At least two of the above are hydrogen atoms. (3) The compound having an azobenzene structure according to (1) or (2), wherein the substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms is a substituted or unsubstituted aliphatic hydrocarbon group having 5 to 35 carbon atoms. (4) The compound having an azobenzene structure according to (1) or (2), wherein the substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms is a substituted or unsubstituted aliphatic hydrocarbon group having 7 to 30 carbon atoms. (5) The above R 1 One of the two and R 2 or -SSZ, wherein one of said groups is independently -Z, -OZ, -CO-OZ, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -SZ, -CO-SZ, -S-CO-Z, or -SSZ. (6) The compound having an azobenzene structure according to any one of (1) to (5), wherein the substituted or unsubstituted aliphatic hydrocarbon group is a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group, or a substituted or unsubstituted alkynyl group. (7) The compound having an azobenzene structure according to any one of (1) to (6), wherein the substituted or unsubstituted aliphatic hydrocarbon group is an unsubstituted aliphatic hydrocarbon group or an aliphatic hydrocarbon group substituted with at least one group selected from an ionic functional group and a nonionic polar functional group. (8) The above R 1 One of the two and R 2 a compound having an azobenzene structure according to any one of (1) to (7), wherein either one of the above is independently a group selected from the following general formulas (2) to (25): [ka] [ka] In the general formulae (2) to (25), n is each independently an integer of 5 to 15; and in the general formulae (4), (6), (8), (20), (22), (24), and (25), X - is a counter anion, and in formulas (10), (12), (14), (16), and (18), Y + is the counter cation.) (9) A vesicle comprising a compound having an azobenzene structure according to any one of (1) to (8) in a lipid bilayer. (10) The vesicle according to (9), wherein the lipid bilayer contains an amphipathic molecule and a compound having an azobenzene structure, and the compound having an azobenzene structure is contained in an amount of 0.01 to 40% by mass relative to a total of 100% by mass of the amphipathic molecule and the compound having an azobenzene structure. (11) The vesicle according to (10), wherein the amphiphilic molecule is a phospholipid. (12) A method for controlling a vesicle structure, comprising irradiating the vesicle according to any one of (9) to (11) with light to cause photoisomerization of a compound having an azobenzene structure, thereby changing the structure of the vesicle. (13) The method for controlling a vesicle structure according to (12), wherein the structural change of the vesicle is division into the inside of the vesicle or division into the outside of the vesicle. Effect of the Invention

[0012] According to the present disclosure, a novel compound having an azobenzene structure and a vesicle containing the compound in a lipid bilayer, which can be used in a method for controlling the structure of a vesicle, are provided. The method for controlling the structure of a vesicle provided by the present disclosure is capable of changing the structure of the vesicle by irradiating it with light. [Brief description of the drawings]

[0013] [Figure 1] Figure 1(a) shows the change in absorption spectrum when a methanol solution of AzoC12-NMe3+ (AzoC12-NMe3+: 0.88 mM) was irradiated with light of a wavelength of 400 nm for 6 minutes, and Figure 1(b) shows the change in absorption spectrum when the solution was irradiated with light of a wavelength of 500 nm for 3 minutes after the irradiation with light of a wavelength of 400 nm. [Diagram 2] The results of HPLC analysis of a methanol solution of AzoC12-NMe3 + (AzoC12-NMe3 +: 0.88 mM) are shown in Figure 2(a). The prepared solution was irradiated with light of 400 nm wavelength for 10 minutes, a portion of the solution was taken, and the results of HPLC analysis are shown in Figure 2(b). After the irradiation with light of 400 nm wavelength, a portion of the solution was irradiated with light of 500 nm wavelength for 3 minutes, and the results of HPLC analysis are shown in Figure 2(c). [Diagram 3] FIG. 3 shows the structure of a compound in which one of R1 and one of R2 in general formula (1) is -O-Me, and the other of R1, the other of R2, and R3 to R10 are hydrogen atoms (the azo group (-N=N-) is in a cis or trans form), as determined by a structural optimization calculation (calculation conditions: DFT B3LYP 6-31G*). [Figure 4] FIG. 4 shows the absorption spectra of the GUV dispersions obtained in Examples 3 to 5 before and after irradiation with light having a wavelength of 400 nm for 1 minute or 10 minutes. [Diagram 5] FIG. 13 is a diagram (micrograph) showing the observation results of Example 7 (division into the inside of the vesicle). [Figure 6] FIG. 13 is a diagram (micrograph) showing the observation results of Example 9 (outward division of vesicles). [Figure 7]FIG. 13 is a diagram (micrograph) showing the observation results of Example 11 (outward division of vesicles). [Figure 8] FIG. 13 is a diagram (microscope photograph) showing the observation results of Example 12 (PS beads are taken up into the inside of the vesicle together with their fragmentation due to membrane deformation). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present invention will be described in detail below.

[0015] <Compounds with an azobenzene structure> The compound having an azobenzene structure according to this embodiment is represented by the following general formula (1). The present inventors have found that a vesicle containing the compound having an azobenzene structure according to this embodiment in a lipid bilayer can have its structure changed by irradiation with light. Although the reason for this is not clear, one of the reasons is that in the compound having an azobenzene structure represented by general formula (1), the azo group (-N=N-) constitutes an eight-membered ring, and therefore the relative positions of the two benzene rings constituting the azobenzene structure and the R 1 and R 2 The inventors hypothesized that this is because the relative angle between the vesicle and the vesicle changes significantly, and this significant structural change causes a change in the fluidity of other components that make up the vesicle (e.g., amphipathic molecules such as phospholipids), which in turn causes a change in the structure of the vesicle.

[0016] [ka]

[0017] (In the general formula (1), R 1 One of the two and R 2 each independently is -Z, -OZ, -CO-OZ, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, -NH-CO-OZ, -O-CO-NH-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -SZ, -CO-SZ, -S-CO-Z, or -SSZ, Z's each independently represent a substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms, and at least one methylene structure in the substituted or unsubstituted aliphatic hydrocarbon group may be replaced by a structure selected from -O-, -S-, -NH-, -CO-, -SS-, -SO2-, -CO-O-, -CO-NH-, and -NH-CO-O-; R 1 On the other hand, R 2 The other and R 3 ~R 10 are each independently a hydrogen atom or a hydrocarbon group having 1 to 24 carbon atoms.

[0018] As mentioned above, R 1 One of the two and R 2 are each independently -Z, -OZ, -CO-OZ, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, -NH-CO-OZ, -O-CO-NH-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -SZ, -CO-SZ, -S-CO-Z, or -SSZ, more preferably -Z, -OZ, -CO-OZ, -O-CO-Z, -CO-NH-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -SZ, -CO-SZ, -S-CO-Z, or -SSZ, further preferably -Z, -OZ, -CO-OZ, -O-CO-Z, -CO-NH-Z, or -NH-CO-Z, and particularly preferably -Z or -OZ. These groups are preferred because they are easy to synthesize and can easily cause the structural change of the vesicle, which will be described later.

[0019] Each Z is independently a substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms. At least one of the methylene structures in the substituted or unsubstituted aliphatic hydrocarbon group may be replaced with a structure selected from -O-, -S-, -NH-, -CO-, -SS-, -SO2-, -CO-O-, -CO-NH-, and -NH-CO-O-. The replacement may be one methylene structure or two or more methylene structures per substituted or unsubstituted aliphatic hydrocarbon group. The number of replaced methylene structures is usually 10 or less, and preferably 2 or less.

[0020] When Z is a substituted aliphatic hydrocarbon group, the substituted aliphatic hydrocarbon group may be a group in which at least one methylene structure in an unsubstituted aliphatic hydrocarbon group is replaced with a structure selected from -O-, -S-, -NH-, -CO-, -SS-, -SO2-, -CO-O-, -CO-NH-, and -NH-CO-O-, or a group in which an end of an unsubstituted aliphatic hydrocarbon group is substituted with a substituent, or a group in which at least one methylene structure in an unsubstituted aliphatic hydrocarbon group is replaced with a structure selected from -O-, -S-, -NH-, -CO-, -SS-, -SO2-, -CO-O-, -CO-NH-, and -NH-CO-O- and the end is substituted with a substituent.

[0021] The substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms is preferably a substituted or unsubstituted aliphatic hydrocarbon group having 5 to 35 carbon atoms, more preferably a substituted or unsubstituted aliphatic hydrocarbon group having 7 to 30 carbon atoms, and particularly preferably a substituted or unsubstituted aliphatic hydrocarbon group having 8 to 25 carbon atoms. From the viewpoint of the balance between the thickness of the lipid bilayer constituting the vesicle and the size of these groups, it is preferable that the number of carbon atoms is within the above range.

[0022] The substituted or unsubstituted aliphatic hydrocarbon group is preferably a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group, or a substituted or unsubstituted alkynyl group. Another preferred embodiment is a substituted or unsubstituted aliphatic hydrocarbon group having at least two, preferably 2 to 5, more preferably 2 to 3 bonds of at least one kind selected from carbon-carbon double bonds and carbon-carbon triple bonds. The carbon-carbon double bond and the carbon-carbon triple bond may be present in the chain constituting the substituted or unsubstituted aliphatic hydrocarbon group, may be present at the terminal, or may be present in the chain and at the terminal.

[0023] Examples of the unsubstituted aliphatic hydrocarbon group include a straight-chain aliphatic hydrocarbon group, a branched chain aliphatic hydrocarbon group, a cyclic aliphatic hydrocarbon group, and an aliphatic hydrocarbon group having a cyclic structure and a chain structure. The substituted aliphatic hydrocarbon group refers to the unsubstituted aliphatic hydrocarbon group substituted with a group or atom other than an aliphatic hydrocarbon.

[0024] Examples of the unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms include a linear or branched alkyl group having 1 to 40 carbon atoms, a linear or branched alkenyl group or an unsaturated double bond-containing group having 2 to 40 carbon atoms, and a cyclic saturated or cyclic unsaturated hydrocarbon group having 3 to 40 carbon atoms.

[0025] Examples of the linear or branched alkyl group having 1 to 40 carbon atoms include a methyl group, an ethyl group, a 1-propyl group, a 1-butyl group, a 1-pentyl group, a 1-hexyl group, a 1-heptyl group, a 1-octyl group, a 1-nonyl group, a 1-decyl group, a 1-undecyl group, a 1-dodecyl group, a 1-tridecyl group, a 1-tetradecyl group, a 1-pentadecyl group, a 1-hexadecyl group, a 1-heptadecyl group, a 1-octadecyl group, a 1-nonadecyl group, a 1-icosyl group, a 1-eicosyl group, a 1-henicosyl group, a 1-hex ... -heneicosyl group, 1-docosyl group, 1-tricosyl group, 1-tetracosyl group, 1-pentacosyl group, 1-hexacosyl group, 1-heptacosyl group, 1-octacosyl group, 1-nonacosyl group, 1-triacontyl group, 1-hentriacontyl group, 1-dotriacontyl group, 1-tritriacontyl group, 1-tetratriacontyl group, 1-pentatriacontyl group, 1-hexatriacontyl group, 1-tetracontyl group, isopropyl group, sec-butyl group, tert-butyl group, iso-butyl, pentan-2-yl, 2-methylbutyl, iso-pentyl, neopentyl, tert-pentyl (1,1-dimethylpropyl), cyamyl, pentan-3-yl, 2-methylpentyl, 3-methylpentyl, iso-hexyl, 1,1-dimethylbutyl (2-methylpentan-2-yl), 3-methylpentan-2-yl, 4-methylpentan-2-yl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, Examples of such alkyl groups include methylbutyl group, thexyl group, 3-methylpentan-3-yl group, 3,3-dimethylbutan-2-yl group, hexane-3-yl group, 2-methylpentan-3-yl group, heptan-4-yl group, 2,4-dimethylpentan-2-yl group, 3-ethylpentan-3-yl group, 4,4-dimethylpentyl group, 4-methylheptan-4-yl group, 4-propylheptan-4-yl group, 2,3,3-trimethylbutan-2-yl group, and 2,4,4-trimethylpentan-2-yl group.

[0026] Examples of linear or branched alkenyl groups or unsaturated double bond-containing groups having 2 to 40 carbon atoms include vinyl groups, allyl groups, propenyl groups, isopropenyl groups, allenyl groups, but-3-en-1-yl groups, crotyl groups, but-3-en-2-yl groups, methallyl groups, buta-1,3-dienyl groups, pent-4-en-1-yl groups, pent-3-en-1-yl groups, pent-2-en-1-yl groups, isopentenyl groups, 2-methylbut-3-enyl groups, Penta-1-yl, pent-4-en-2-yl, prenyl, 2-methyl-but-2-en-1-yl, pent-3-en-2-yl, 2-methyl-but-3-en-2-yl, pent-1-en-3-yl, penta-2,4-dien-1-yl, penta-1,3-dien-1-yl, penta-1,4-dien-3-yl, iso-prenyl (2-methyl-but-1,3-dien-1-yl), penta-2,4-dien-2-yl hex-5-en-1-yl, hex-4-en-1-yl, hex-3-en-1-yl, hex-2-en-1-yl, 4-methyl-pent-4-en-1-yl, 3-methyl-pent-4-en-1-yl, 2-methyl-pent-4-en-1-yl, hex-5-en-2-yl, 4-methyl-pent-3-en-1-yl, 3-methyl-pent-3-en-1-yl, 2,3-dimethyl-but-2-en-1 Examples of the aryl group include a 2-methylpent-4-en-2-yl group, a 3-ethylpent-1-en-3-yl group, a hexa-3,5-dien-1-yl group, a hexa-2,4-dien-1-yl group, a 4-methylpenta-1,3-dien-1-yl group, a 2,3-dimethyl-buta-1,3-dien-1-yl group, a hexa-1,3,5-trien-1-yl group, a 2-(cyclopentadienyl)propan-2-yl group, and a 2-(cyclopentadienyl)ethyl group.

[0027] Examples of the cyclic saturated or cyclic unsaturated hydrocarbon group having 3 to 40 carbon atoms include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclopentenyl group, a cyclopentadienyl group, a dimethylcyclopentadienyl group, an n-butylcyclopentadienyl group, an n-butyl-methylcyclopentadienyl group, a tetramethylcyclopentadienyl group, a 1-methylcyclopentyl group, a 1-allylcyclopentyl group, a 1-benzylcyclopentyl group, a cyclohexyl group, a cyclohexenyl group, a cyclohexadienyl group, a 1-methylcyclohexyl group, a 1-allylcyclohexyl group, a 1-benzylcyclohexyl group, a cycloheptyl group, a cycloheptenyl group, a cycloheptatrienyl group, a 1-methylcycloheptyl group, a 1-allylcycloheptyl group, a 1-benzylcycloheptyl group, Cyclooctyl, cyclooctenyl, cyclooctadienyl, cyclooctatrienyl, 1-methylcyclooctyl, 1-allylcyclooctyl, 1-benzylcyclooctyl, 4-cyclohexyl-tert-butyl, norbornyl, norbornenyl, norbornadienyl, 2-methylbicyclo[2.2.1]heptan-2-yl, 7-methylbicyclo[2. 2.1]heptan-7-yl group, bicyclo[2.2.2]octan-1-yl group, bicyclo[2.2.2]octan-2-yl group, 1-adamantyl group, 2-adamantyl group, 1-(2-methyladamantyl), 1-(3-methyladamantyl), 1-(4-methyladamantyl), 1-(3,5-dimethyladamantyl), 1-(3,5,7-trimethyladamantyl), and the like.

[0028] Examples of the substituted aliphatic hydrocarbon group having 1 to 40 carbon atoms include groups obtained by substituting the unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms. Examples of the substituted aliphatic hydrocarbon group include aliphatic hydrocarbon groups substituted with at least one group selected from an ionic functional group and a nonionic polar functional group. In other words, in one preferred embodiment, the substituted or unsubstituted aliphatic hydrocarbon group is an unsubstituted aliphatic hydrocarbon group, or an aliphatic hydrocarbon group substituted with at least one group selected from an ionic functional group and a nonionic polar functional group.

[0029] The at least one group selected from the ionic functional group and the nonionic polar functional group is not particularly limited, but is preferably at least one group selected from the following formulae (A1) to (A23). In the present invention, the wavy line at the end of the group indicates a bonding site with another atom.

[0030] [ka]

[0031] [ka]

[0032] (In the formulae (A2), (A4), (A6), (A18), (A20), (A22) and (A23), X - is a counter anion, and in formulae (A8), (A10), (A12), (A14) and (A16), Y + is the counter cation.)

[0033] The at least one group selected from the ionic functional groups and the nonionic polar functional groups is more preferably at least one group selected from the formulae (A2), (A4), (A6), (A8), (A10), (A12), (A14), (A16), (A18), (A20), (A22), and (A23), and particularly preferably at least one group selected from the formulae (A2), (A6), (A10), (A12), (A14), and (A16).

[0034] Z is a substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms, and in one preferred embodiment, the aliphatic hydrocarbon group portion is a straight-chain alkyl group. In addition, the at least one group selected from ionic functional groups and nonionic polar functional groups is not particularly limited, but is preferably at least one group selected from the above formulae (A1) to (A23). In one preferred embodiment, the end of the straight-chain alkyl group is substituted with at least one group selected from ionic functional groups and nonionic polar functional groups. Specifically, Z is preferably at least one group selected from the following general formulae (B1) to (B24). As described above, R 1 One of the two and R 2 is particularly preferably -Z or -OZ. 1 One of the two and R 2 It is particularly preferable that one of R is independently a group selected from the following general formulae (B1) to (B24) and the following general formulae (2) to (25). 1 One of the two and R 2 It is more preferable that either one of R is independently a group selected from the following general formulas (2) to (25). 1 One of the two and R 2 may be different groups, but is preferably the same group from the viewpoint of ease of synthesis.

[0035] [ka]

[0036] [ka]

[0037] In the general formulae (B1) to (B24), n is each independently an integer of 5 to 15; and in the groups (B3), (B5), (B7), (B19), (B21), (B23) and (B24), X - is a counter anion, and in groups (B9), (B11), (B13), (B15) and (B17), Y+ is the counter cation.)

[0038] [ka]

[0039] [ka]

[0040] In the general formulae (2) to (25), n is each independently an integer of 5 to 15; and in the general formulae (4), (6), (8), (20), (22), (24), and (25), X - is a counter anion, and in formulas (10), (12), (14), (16), and (18), Y + is the counter cation.)

[0041] Examples of the counter anion include halide ions (e.g., F - , Cl - , Br - , I - ) As the counter cation, for example, an alkali metal ion (e.g., Li + , Na + , K + The counter anion and counter cation may be a divalent or higher ion. For example, calcium ion (Ca 2+ If Ca is the counter cation, 2+ ) 1 / 2 " is Y + is equivalent to.

[0042] Each n is independently an integer of 5 to 15, preferably an integer of 7 to 15, and more preferably an integer of 10 to 15.

[0043] As mentioned above, R 1 On the other hand, R 2 The other and R 3 ~R10 are each independently a hydrogen atom or a hydrocarbon group having 1 to 24 carbon atoms, preferably a hydrogen atom or a hydrocarbon group having 1 to 20 carbon atoms, more preferably a hydrogen atom or a hydrocarbon group having 1 to 15 carbon atoms, even more preferably a hydrogen atom or a hydrocarbon group having 1 to 10 carbon atoms, and particularly preferably a hydrogen atom or a hydrocarbon group having 1 to 6 carbon atoms.

[0044] R 1 The other and R 2 and the other is preferably a hydrogen atom, and more preferably both are hydrogen atoms. 3 ~R 6 At least two of R 7 ~R 10 At least two of R are preferably hydrogen atoms. 3 ~R 6 At least three of R 7 ~R 10 More preferably, at least three of R 3 ~R 6 are all hydrogen atoms, 7 ~R 10 It is particularly preferred that all of R 1 On the other hand, R 2 The other and R 3 ~R 10 In one particularly preferred embodiment, all of are hydrogen atoms.

[0045] Examples of the hydrocarbon group having 1 to 24 carbon atoms include an aliphatic hydrocarbon group having 1 to 24 carbon atoms and an aromatic hydrocarbon group having 6 to 24 carbon atoms, with an aliphatic hydrocarbon group having 1 to 24 carbon atoms being preferred from the viewpoint of easy mixing with amphipathic molecules such as phospholipids. Furthermore, as the hydrocarbon group having 1 to 24 carbon atoms, a hydrocarbon group having 1 to 15 carbon atoms is preferred, a hydrocarbon group having 1 to 10 carbon atoms is more preferred, and a hydrocarbon group having 1 to 6 carbon atoms is particularly preferred.

[0046] Examples of the aromatic hydrocarbon group having 6 to 24 carbon atoms include a phenyl group, a tolyl group, a naphthyl group, a biphenyl group, and an anthracenyl group.

[0047] Examples of the aliphatic hydrocarbon group having 1 to 24 carbon atoms include a linear or branched alkyl group having 1 to 24 carbon atoms, a linear or branched alkenyl group or an unsaturated double bond-containing group having 2 to 24 carbon atoms, and a cyclic saturated or cyclic unsaturated hydrocarbon group having 3 to 24 carbon atoms.

[0048] Examples of the linear or branched alkyl group having 1 to 24 carbon atoms include those having 24 or less carbon atoms among the groups listed above as linear or branched alkyl groups having 1 to 40 carbon atoms. Examples of the linear or branched alkenyl group or unsaturated double bond-containing group having 2 to 24 carbon atoms include those having 24 or less carbon atoms among the groups listed above as linear or branched alkenyl group or unsaturated double bond-containing group having 2 to 40 carbon atoms. Examples of the cyclic saturated or cyclic unsaturated hydrocarbon group having 3 to 24 carbon atoms include those having 24 or less carbon atoms among the groups listed above as cyclic saturated or cyclic unsaturated hydrocarbon groups having 3 to 40 carbon atoms.

[0049] As the hydrocarbon group having 1 to 24 carbon atoms, a methyl group, an ethyl group, a 1-propyl group, or an iso-propyl group is particularly preferred.

[0050] Specific examples of the compound having an azobenzene structure according to this embodiment include the compounds represented by the following formulae (C1) to (C3) which are synthesized in the examples described later. In the present invention, the compound represented by formula (C1) is referred to as AzoC 12 The compound represented by formula (C2) is also referred to as AzoC 12 -NMe3 + The compound represented by formula (C3) is also referred to as AzoC 16 Also written as.

[0051] [ka]

[0052] The method for synthesizing the compound having an azobenzene structure according to the present embodiment is not particularly limited, and the compound can be synthesized by a conventionally known organic synthesis method, such as the method described in the Examples or the method described in W. Moormann et al., “Solvent-free synthesis of diazocine”, Synthesis 49:3471 (2017).

[0053] (Vesicles) The vesicle according to the present embodiment contains the compound having the azobenzene structure according to the present embodiment in the lipid bilayer. By irradiating the vesicle according to the present embodiment with light as described below, it is possible to control the structure of the vesicle, such as splitting the vesicle into the inside or the outside.

[0054] The lipid bilayer contains a compound having an azobenzene structure, but also usually contains amphipathic molecules. That is, it is preferable that the lipid bilayer contains an amphipathic molecule and a compound having an azobenzene structure. In addition, it is preferable that the compound having an azobenzene structure is contained in an amount of 0.01 to 40% by mass, more preferably 0.8 to 25% by mass, and particularly preferably 5 to 15% by mass, relative to the total 100% by mass of the amphipathic molecule and the compound having an azobenzene structure. The vesicle according to this embodiment is useful compared to KAON12, which requires a large amount of use, because the structure of the vesicle can be controlled even when the amount of the compound having an azobenzene structure relative to the total 100% by mass of the amphipathic molecule and the compound having an azobenzene structure is small.

[0055] Examples of the amphipathic molecule include phospholipids, glycolipids, sterols, cationic lipids, etc. From the viewpoint of forming a stable bilayer membrane (lipid bilayer), it is preferable that the amphipathic molecule is a phospholipid.

[0056] When the amphiphilic molecule is a phospholipid, the compound having an azobenzene structure is preferably 40 mol% or less, more preferably 30 mol% or less, even more preferably 20 mol% or less, and particularly preferably 15 mol% or less, relative to the total of 100 mol% of the phospholipid and the compound having an azobenzene structure. Also, the compound having an azobenzene structure is preferably 0.01 mol% or more, more preferably 0.1 mol% or more, even more preferably 0.5 mol% or more, and particularly preferably 1 mol% or more.

[0057] Examples of phospholipids include DOPC (1,2-dioleoyl sn-glycero-3-phosphocholine), DOPA (1,2-dioleoyl-sn-glycero-3-phosphate sodium salt), DPPC (1,2-dipalmitoyl sn-glycero-3-phosphocholine), DMPC (1,2-ministryl sn-glycero-3-phosphocholine), DPPG (1,2-dipalmitoyl sn-glycero-3-phosphatidylglycerol), and POPC (1,2-dipalmitoyl-oleyl sn-glycero-3-phosphocholine).

[0058] Examples of glycolipids include glyceroglycolipids and sphingoglycolipids. Examples of sterols include cholesterol and phytosterols (e.g., sitosterol, stigmasterol, fucosterol, spinasterol, brassicasterol, etc.).

[0059] An example of the cationic lipid is DOTAP (1,2-dioleoyl-3-trimethylammonium-propane chloride).

[0060] The lipid bilayer constituting the vesicle may contain components other than amphipathic molecules and compounds having an azobenzene structure. Examples of components other than amphipathic molecules and compounds having an azobenzene structure include drugs, detection drugs, and hydrophobic lipid molecules such as cholesterol and cholic acid.

[0061] The vesicle according to the present embodiment usually contains a liquid (water, a solution such as a buffer solution containing water, etc.) inside the lipid bilayer (inside the vesicle), but may contain other components depending on the intended use of the vesicle. Examples of other components contained in the vesicle include drugs, detection agents, nucleic acids (DNA, RNA), proteins, and sugar chains.

[0062] The size of the vesicle may be appropriately set depending on the purpose of use of the vesicle and is not particularly limited, but is, for example, 500 nm to 200 μm, preferably 1 to 150 μm, more preferably 3 to 100 μm, and particularly preferably 5 to 50 μm.

[0063] The method for producing vesicles is not particularly limited except that the compound having the azobenzene structure described above is used as a material for forming the lipid bilayer, and can be performed according to conventional artificial vesicle production methods such as the method described in the Examples or the method described in T. Muraoka et al., “A synthetic ion channel with anisotropic ligand response”, Nature Communication 11:2924 (2020).

[0064] <Method for controlling vesicle structure> In the method for controlling the structure of a vesicle according to this embodiment, the vesicle according to this embodiment described above is irradiated with light to cause photoisomerization of a compound having an azobenzene structure, thereby changing the structure of the vesicle.

[0065] The method for controlling the structure of a vesicle is usually carried out in a liquid, preferably in an aqueous solution. The aqueous solution contains water, and also contains components for forming a buffer solution and components for forming a liquid medium. There is no particular limit to the pH of the aqueous solution, but it is usually preferably 5 to 9, more preferably 6 to 8, and particularly preferably 6.5 to 7.5. pH 7 is one of the preferred embodiments.

[0066] The method for controlling the vesicle structure is usually carried out at 100° C. or lower, preferably at 5 to 60° C., and more preferably at 20 to 40° C. In one preferred embodiment, the temperature is room temperature (20 to 30° C.) or around body temperature (35 to 38° C.).

[0067] The method for controlling the structure of a vesicle may be carried out under normal pressure (atmospheric pressure), under pressure, or under reduced pressure, but is usually carried out under normal pressure. The atmosphere in which the method for controlling the structure of a vesicle is carried out is not particularly limited, and may be carried out under an atmospheric (air) atmosphere, an inert gas (e.g., rare gas, nitrogen) atmosphere, an oxygen atmosphere, a carbon dioxide atmosphere, or a mixed gas atmosphere.

[0068] The method for controlling the structure of a vesicle according to the present embodiment can control the dynamic behavior of a vesicle by light irradiation. One of the reasons for this is that the compound having an azobenzene structure represented by general formula (1) contained in the lipid bilayer of the vesicle has an azo group (-N=N-) that constitutes an eight-membered ring, and therefore the relative positions of the two benzene rings that constitute the azobenzene structure and the R 1 and R 2 The inventors presume that this is because the relative angle between the vesicle and the vesicle changes significantly, and this significant structural change causes a change in the fluidity of other components constituting the vesicle (e.g., amphipathic molecules such as phospholipids), which can cause a structural change in the vesicle. The structural change in the vesicle is preferably division into the inside of the vesicle or division into the outside of the vesicle. Here, division into the inside of the vesicle means that by irradiating a vesicle with light, the lipid bilayer membrane penetrates toward the inside of the vesicle, and another vesicle is formed inside the vesicle. Moreover, division into the outside of the vesicle means that by irradiating a vesicle with light, the lipid bilayer membrane buds toward the outside of the vesicle, and another vesicle is formed outside the vesicle.

[0069] In one preferred embodiment, the light irradiation is light containing at least visible light having a wavelength of 367 to 420 nm from the viewpoint of efficiently performing photoisomerization of a compound having an azobenzene structure. In another preferred embodiment, the light irradiation is light containing at least visible light having a wavelength of 460 to 530 nm from the viewpoint of efficiently performing photoisomerization of a compound having an azobenzene structure. When the azo group (-N=N-) of a compound having an azobenzene structure is a cis-form, photoisomerization can be efficiently performed with visible light having a wavelength of 367 to 420 nm. On the other hand, when the azo group (-N=N-) of a compound having an azobenzene structure is a trans-form, photoisomerization can be efficiently performed with visible light having a wavelength of 460 to 530 nm. The wavelength of the light irradiation can be appropriately set depending on whether the azo group (-N=N-) is a cis-form or a trans-form, etc.

[0070] Since visible light, which is less harmful to living organisms, can be used as light irradiation, the method for controlling the structure of a vesicle according to the present embodiment can be used for various applications. The method for controlling the structure of a vesicle according to the present embodiment may be performed in a living organism (e.g., humans, non-human mammals, birds, reptiles, fish, crustaceans, insects) or outside the living organism (in vitro).

[0071] The vesicle structure control method according to the present embodiment can utilize the structural change of the vesicle to take in a substance present outside the vesicle into the vesicle or release a substance present inside the vesicle to the outside, and can therefore be used for various applications. Specific examples of applications include the use in research in the fields of nucleic acid introduction, drug delivery systems, and regenerative medical materials. EXAMPLES

[0072] The present embodiment will be described below with reference to examples, but the present disclosure is not limited to these examples.

[0073] [Example 1] AzoC is synthesized by the method shown in the scheme below. 12 -NH2 and AzoC12 -NMe3 + was synthesized.

[0074] [ka]

[0075] (Synthesis of 1-azido-12-bromododecane (product 3)) [ka]

[0076] After replacing the atmosphere in a two-necked eggplant flask with nitrogen, 1,12-dibromododecane (compound 2) (3.0 g, 9.14 mmol) was added and dissolved in dry DMF (10 ml). NaN3 (198 mg, 3.05 mmol) was then added and the mixture was stirred at 60°C for 19 hours. 100 ml of water was then added, and the mixture was extracted three times with 30 ml of ethyl acetate, and washed once with 200 ml of saturated saline. Na2SO4 was added to the separated organic phase, filtered, and concentrated under reduced pressure. Product 3 (820 mg, 31%) was obtained as a pale yellow liquid by silica gel column chromatography (hexane).

[0077] The analytical results of product 3 were as follows: TLC R f (Merck 60 F254, Hexane): 0.38 1 H NMR (400 MHz, CDCl3 with TMS): δ 3.38 (t, J = 7.1 Hz, 2H), 3.24 (t, J = 7.1 Hz, 2H), 1.84 (quin, J = 7.3 Hz, 2H), 1.58 (quin, J = 7.1 Hz, 2H), 1.28 (m, 16H)ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ 26.7, 28.2, 28.7, 28.8, 29.1, 29.4, 29.5, 32.8, 33.8, 51.4 ppm ESI MS (positive mode): m / z: C 12 H 24 Calculated value for BrN3: [M + H] + 290.1232; Actual value: 290.1230

[0078] (Synthesis of (1S,2S)-1,2-bis(2-nitrophenyl)ethane-1,2-diol (product 5)) [ka]

[0079] After replacing the atmosphere in a two-necked eggplant flask with nitrogen, compound 4 (5.0 g, 33.1 mmol) was placed in the flask and dissolved in THF / Water = 3 / 1 (40 ml). After degassing with nitrogen bubbling for 30 minutes, PBu3 (6.7 g, 33.1 mmol) was added and the mixture was stirred at room temperature for 12 hours. Water was then added, and the mixture was extracted three times with methylene chloride. The separated organic phase was washed once with saturated saline. Na2SO4 was added to the organic phase, filtered, and concentrated under reduced pressure. Product 5 (2.8 g, 56%) was obtained as a pale yellow solid by reprecipitation (ethanol).

[0080] The analytical results of product 5 were as follows: TLC R f (Merck 60 F254, CH2Cl2): 0.13 1 H NMR (400 MHz, CDCl3 with TMS): δ 7.78 (2H, t, J = 8.0 Hz), 7.62 (2H, t, J = 7.2 Hz), 7.44 (2H, t, J = 7.2 Hz), 5.59 (2H, s), 3.49 (2H, s,) ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ 71.9, 124.5, 129.0, 129.4, 133.2, 134.7, 148.6 ppm ESI MS (positive mode): m / z: C 14 H 12Calculated for N2O6: [M + Na]+ 327.0593; Found: 327.0592

[0081] (Synthesis of (1S,2S)-1,2-bis(2-aminophenyl)ethane-1,2-diol (Product 6)) [ka]

[0082] After replacing the atmosphere in a two-necked eggplant flask with nitrogen, product 5 (300 mg, 0.98 mmol) was placed in the flask and dissolved in EtOH / Water = 10 / 1 (40 ml). Fe (542 mg, 9.86 mmol) was then added, followed by NH4Cl (106 mg, 1.97 mmol), and the mixture was heated under reflux for 2 hours. After the reaction was complete, the mixture was cooled to room temperature. Water was added, and the mixture was extracted three times with methylene chloride. The separated organic phase was washed once with saturated saline. Na2SO4 was added to the organic phase, which was then filtered and concentrated under reduced pressure. Product 6 (150 mg, 63%) was obtained as a white solid by recrystallization (ethanol).

[0083] The analytical results of product 6 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 1): 0.13 1 H NMR (400 MHz, CDCl3 with TMS): δ 7.02 (2H, t, J = 7.6 Hz), 6.91 (2H, dd, J = 7.6 Hz, J = 1.6 Hz), 6.63 (2H, t, J = 7.6 Hz), 6.57 (2H, dd, J = 7.5 Hz, J = 1.0 Hz) 5.06 (2H, s), 4.03 (6H, br) ppm ESI MS (positive mode): m / z: C 14 H 16 Calculated value for N2O2: [M + H] + 245.1290; Actual value: 245.1288

[0084] (Synthesis of 2,2'-((1R,2R)-1,2-bis((12-azidododecyl)oxy)ethane-1,2-diyl)dianiline (Product 7)) [ka]

[0085] After replacing the atmosphere in a two-necked eggplant flask with nitrogen, NaH (60%, dispersed in liquid paraffin, 65 mg, 1.64 mmol) was added and washed three times with dry hexane. After that, it was vacuum dried and dry DMF (2 ml) was added. After that, it was cooled on ice and a dry DMF solution (3 ml) of product 6 (100 mg, 0.41 mmol) was added dropwise over 7 minutes. After the dropwise addition, it was stirred at room temperature for 30 minutes. Then, a dry DMF (1 ml) solution of product 3 (261 mg, 0.90 mmol, 2.05 mmol) was added dropwise over 5 minutes. After the dropwise addition, it was stirred at room temperature for 5 hours. Water was added to inactivate NaH, and it was extracted three times with ethyl acetate. Na2SO4 was added to the separated organic phase, filtered, concentrated under reduced pressure, and then product 7 (203 mg, 75%) was obtained as a pale yellow oily liquid by silica gel column chromatography (ethyl acetate / hexane = 1 / 4).

[0086] The analytical results of product 7 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 4): 0.39 1 H NMR (400 MHz, CDCl3 with TMS): δ 7.21 (2H, dd, J = 7.6 Hz, J = 1.6 Hz), 7.08 (2H, td, J = 7.6 Hz, J = 1.7 Hz), 6.75 (2H, t, J = 7.6 Hz), 6.65 (2H, dd, J = 8.0 Hz, J = 0.9 Hz) 4.59 (2H, s), 4.21 (4H, br), 3.32 (2H, m), 3.25 (4H, t, J = 6.9 Hz), 3.03 (2H, m), 1.60 (4H, quin, J = 7.3 Hz), 1.61-1.02 (36H, m)ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ 26.0, 26.9, 29.0, 29.3, 29.4, 29.7, 51.6, 69.7, 82.7, 116.6, 118.2, 125.6, 128.2, 129.2, 146,0 ppm ESI MS (positive mode): m / z: C 38 H 62 Calculated value for N8O2: [M + H] + 663.5074; Actual value: 663.5073

[0087] (Synthesis of (11R,12R,Z)-11,12-bis((12-azidododecyl)oxy)-11,12-dihydrodibenzo[c,g][1,2]diazocine (Product 8)) [ka]

[0088] After replacing the atmosphere in a two-necked eggplant flask containing product 7 (460 mg, 0.69 mmol), product 7 was dissolved in dry CHCl (10 ml). PIDA (iodobenzene diacetate) (447 mg, 1.39 mmol) was then added and stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure and subjected to column chromatography (ethyl acetate / hexane = 1 / 4) to obtain product 8 (171 mg, 37%) as a yellow oily liquid.

[0089] The analytical results of product 8 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 4): 0.48 1H NMR (400 MHz, CDCl3 with TMS): δ 7.50 (2H, d, J = 7.6 Hz), 7.21 (2H, td, J = 7.5 Hz, J = 1.3 Hz), 7.16 (2H, t, J = 7.5 Hz), 6.95 (2H, dd, J = 7.5 Hz, J = 1.0 Hz) 4.58 (2H, s), 3.76 (2H, q), 3.62 (2H, q, J = 6.3 Hz), 3.25 (4H, t), 1.66-1.58 (8H, m), 1.28 (32H, m) ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ 26.4, 26.9, 27.0, 29.0, 29.3, 29.6, 29.73, 29.77, 29.8, 30.3, 32.8, 45.3, 51.6, 72.7, 79.6, 119.9, 127.50, 127.57, 127.7, 152.8 ppm ESI MS (positive mode): m / z: C 38 H 58 Calculated value of N8O2: [M + Na] + 681.4580; Actual value: 681.4582

[0090] (12,12'-(((11R,12R,Z)-11,12-dihydrodibenzo[c,g][1,2]diazocin-11,12-diyl)bis(oxy))bis(dodecan-1-amine), (Product: AzoC 12 Synthesis of -NH2) [ka]

[0091] After replacing the atmosphere in a two-necked recovery flask containing product 8 (170 mg, 0.26 mmol), product 8 was dissolved in dry THF (5 ml). PPh3 (271 mg, 1.03 mmol) was added thereto and stirred at room temperature for 15 hours. Water (1 ml) was then added and stirred at room temperature for an additional 10 hours. The reaction solution was concentrated under reduced pressure and purified by column chromatography (NH silica, CHCl3 containing 5% MeOH (methanol)). 12 -NH2 (81 mg, 51%) was obtained as a yellow solid.

[0092] AzoC 12 The results of the -NH2 analysis were as follows: TLC R f (NHsilica, CHCl3 containing 5% MeOH): 0.50 1 H NMR (400 MHz, CDCl3 with TMS): δ 7.50 (2H, d, J = 7.6 Hz,), 7.21 (2H, t, J = 7.5 Hz), 7.16 (2H, t, J = 7.5 Hz), 6.95 (2H, dd, J = 7.5 Hz, J = 1.0 Hz) 4.58 (2H, s), 3.76 (2H, q), 3.62 (2H, q, J = 6.3 Hz), 2.67 (4H, t, J = 6.4 Hz), 1.66 (4H, quin, J = 7.1 Hz), 1.41 (8H, m), 1.28 (32H, s) ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ 26.3, 27.0, 29.60, 29.64, 29.76, 30.3, 33.94, 42.4, 62.4, 72.6, 79.6, 119.9, 127.4, 127.5, 127.7, 152.8 ppm ESI MS (positive mode): m / z: C 38 H 62 Calculated value for N4O2: [M + H] + 607.4951; Actual value: 607.4948

[0093] (12,12'-(((11R,12R,Z)-11,12-dihydrodibenzo[c,g][1,2]diazocin-11,12-diyl)bis(oxy))bis(N,N,N-trimethyldodecane-1-ammonium), (Product: AzoC 12 -NMe3 + ) synthesis [ka]

[0094] AzoC 12 After replacing the nitrogen gas in a two-necked eggplant flask containing -NH2 (45 mg, 0.074 mmol), AzoC was added to dry MeOH (methanol) (5 ml). 12 The -NH2 was dissolved. Then, MeI (126 mg, 0.89 mmol) was added, followed by K2CO3 (123 mg, 0.89 mmol) dried at 120 °C for 20 min, and the mixture was stirred for 24 hours. The reaction solution was concentrated under reduced pressure and purified by HPLC to obtain AzoC 12 -NH2 and AzoC 12 -NMe3 + (63 mg, 90%) was obtained.

[0095] HPLC purification was carried out under the following conditions. Model: JASCO HPLC Detector: UV-4075 UV / VIS detector Detection wavelength: 425nm Column: YMC Triat C18 column Mobile phase: Water / acetonitrile (volume ratio) containing 0.1 vol% TFA (trifluoroacetic acid), 0-3 min: 70 / 30, 3 min to less than 18 min: gradient, 18-23 min: 0 / 100, 23 min to less than 30 min: gradient, 30-35 min: 70 / 30 Flow rate: 18.9mL / min

[0096] AzoC 12 -NMe3 + The results of the analysis were as follows: TLC R f(NHsilica, CHCl3 containing 5% MeOH): 0.01 1 H NMR (400 MHz, CD3OD with TMS): δ 7.47 (2H, m), 7.27 (2H, t, J = 7.5 Hz), 7.18 (2H, t, J = 7.5 Hz), 6.94 (2H, d, J = 7.5 Hz) 4.61 (2H, s, CH2), 3.37 (4H, m), 3.15 (18H, s), 1.78 (4H, m), 1.39-1.35 (40H, m) ppm ESI MS (positive mode): m / z: C 44 H 76 Calculated value of I2N4O2: [MI] + 819.5007, Actual value [MI] + 819.0000

[0097] (AzoC 12 -NMe3 + Changes in absorption spectrum due to photoisomerization of AzoC 12 -NMe3 + In order to confirm the photoisomerization of the compound, the absorption spectrum was measured. AzoC 12 -NMe3 + Dissolve in MeOH (methanol) and add AzoC 12 -NMe3 + A solution containing 0.88 mM of was prepared. Figure 1(a) shows the change in absorption spectrum when the prepared solution was irradiated with light of 400 nm wavelength for 6 minutes, and Figure 1(b) shows the change in absorption spectrum when the solution was irradiated with light of 500 nm wavelength for 3 minutes after the irradiation with light of 400 nm wavelength. The light irradiation was performed using a 300 W xenon light source MAX-303 (manufactured by Asahi Spectroscopy).

[0098] Figures 1(a) and 1(b) show that the absorption spectrum has an isosbestic point and changes reversibly. 12 -NMe3 +It was suggested that the azo group (-N=N-) of isomerized by light. From the change in wavelength of the absorption spectrum, it was suggested that isomerization from the cis form to the trans form occurs upon light irradiation in Figure 1(a), and that isomerization from the trans form to the cis form occurs upon light irradiation in Figure 1(b). Furthermore, from Figure 1, the absorption spectrum shifts to the longer wavelength side than KAON12, and the absorption wavelength of the cis form is well within the visible light wavelength of 400 nm or more, suggesting that it has visible light responsiveness.

[0099] (AzoC 12 -NMe3 + Analysis of photoisomerization of by high performance liquid chromatography (HPLC) AzoC 12 -NMe3 + HPLC analysis was carried out to analyze the photoisomerization of . AzoC 12 -NMe3 + Dissolve in MeOH and add AzoC 12 -NMe3 + A solution containing 0.88 mM of was prepared. A portion of the prepared solution was taken and subjected to HPLC analysis. The results are shown in Figure 2(a).

[0100] The prepared solution was irradiated with light having a wavelength of 400 nm for 10 minutes, and a portion of the solution was taken and analyzed by HPLC. The results are shown in Figure 2(b). After the irradiation with light having a wavelength of 400 nm, the solution was irradiated with light having a wavelength of 500 nm for 3 minutes, and a portion of the solution was taken and analyzed by HPLC. The results are shown in Figure 2(c). Irradiation with light was performed using a 300 W xenon light source MAX-303 (manufactured by Asahi Spectroscopy).

[0101] The HPLC analysis was carried out under the following conditions. Model: JASCO HPLC Detector: UV-4075 UV / VIS detector Detection wavelength: 425nm Column: YMC Triat C18 column Mobile phase: Water / acetonitrile (volume ratio) containing 0.1 vol% TFA (trifluoroacetic acid), 0-2 min: 70 / 30, 2 min to less than 18 min: gradient, 18-27 min: 0 / 100, 27 min to less than 32 min: gradient, 32 min to 35 min: 70 / 30 Flow rate: 1.0mL / min

[0102] AzoC before light irradiation 12 -NMe3 + More than 99% of AzoC was cis-isomer, and 66% was converted to trans-isomer by 10 min of irradiation at 400 nm. 12 -NMe3 + was again >99% cis.

[0103] (AzoC 12 -NMe3 + (Structure of In the general formula (1), R 1 One of the two and R 2 One of the groups is -O-Me, and the other is R 1 On the other hand, R 2 The other and R 3 ~R 10 The structure of a compound in which R is a hydrogen atom (the azo group (-N=N-) is in either cis or trans form) is shown in Figure 3. Figure 3 shows a side view of the carbon-carbon bond between the carbon atoms bonded to the oxygen atom, looking perpendicular to the paper, and a top view looking from the two methyl groups toward the N=N bond. The angle shown in the side view indicates the dihedral angle between the two C-O bonds. That is, in general formula (1), 1 One of the two and R 2 One of the groups is -O-Me, and the other is R 1 On the other hand, R 2 The other and R 3 ~R 10 It was suggested that the three-dimensional structure of compounds in which the is a hydrogen atom changes significantly upon photoisomerization.

[0104] AzoC 12 -NMe3+ and other compounds synthesized in the examples (AzoC 12 -NH2, AzoC 16 It was also suggested that the three-dimensional structure of compounds having an azobenzene structure represented by general formula (1), such as 1,2-diphenylphosphine (1,2-diphenylphosphine) and 1,2-diphenylphosphine (1,2-diphenylphosphine)-1,3-tri ...

[0105] [Example 2] AzoC is synthesized by the method shown in the scheme below. 16 was synthesized.

[0106] [ka]

[0107] (Synthesis of 1,2-bis(2-nitrophenyl)ethane (product 12)) [ka]

[0108] After placing the flask in a nitrogen atmosphere, 2-nitrotoluene (10.0 g, 75.0 mmol) was added and dissolved in dry THF (250 ml). The flask was placed in an ice bath, potassium tert-butoxide (10.0 g, 89.0 mmol) was added, and the mixture was stirred at 0°C for 5 minutes. Br2 (4.0 g, 25 mmol) was then added and the mixture was stirred for another 15 minutes. After stirring, water was added, and the mixture was extracted three times with methylene chloride. The combined organic phase was washed once with saturated saline. Na2SO4 was added to the separated organic phase, filtered, concentrated under reduced pressure, and recrystallized (ethanol) to obtain product 12 (9.75 g, 97%) as a pale yellow solid.

[0109] The analytical results of product 12 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 3): 0.38 (raw material is 0.5) 1H NMR (400 MHz, CDCl3 with TMS): δ = 7.96 (2H, dd, J = 8.2 Hz, J = 1.2 Hz), 7.54 (2H, pseudo t), 7.42 (2H, dd, J = 7.7 Hz, J = 1.3 Hz), 7.38 (2H, pseudo t), 3.25 (4H, s)ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ 57.1, 84.6, 116.2, 118.2, 121.9, 128.7, 130.0, 145.7 ppm ESI MS (positive mode): m / z: C 14 H 12 Calculated value for N2O4: [M + H] + 273.0875; Actual value: 273.0878

[0110] (Synthesis of (Z)-11,12-dihydrodibenzo[c,g][1,2]diazocine (product 13)) [ka]

[0111] A two-necked eggplant flask containing product 12 (9.75 g, 35 mmol) was replaced with nitrogen, and product 12 was dissolved in EtOH (500 ml). Zn (22 g, 350 mmol) was then added. 0.2 M Ba(OH)2·8H2O aq. (570 ml) was added and the mixture was heated to reflux for 3 hours. After removing Zn by filtration through Celite, an aqueous solution of ammonium chloride was added, and the mixture was extracted three times with ethyl acetate and washed once with saturated saline. Na2SO4 was added to the separated organic phase, filtered, concentrated under reduced pressure, and then column chromatography (ethyl acetate / hexane = 1 / 3) was used to obtain product 13 (2.93 g, 40%) as a yellow solid.

[0112] The analytical results of product 13 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 3): 0.56 1 H NMR (400 MHz, CDCl3 with TMS): δ = 7.13 (2H, pseudo t), 7.02 (2H, pseudo t), 6.98 (2H, d, J = 7.7 Hz), 6.83 (2H, d, J = 7.8 Hz), 2.87 (4H, m) ppm 13 C NMR (400 MHz, CDCl3 with TMS): δ = 155.49, 129.60, 128.08, 127.01, 126.66, 118.70, 31.65 ppm ESI MS (positive mode): m / z: C 14 H 12 Calculated value of N2: [M + H] + 209.1079; Actual value: 209,1079

[0113] (Synthesis of (Z)-11-bromo-11,12-dihydrodibenzo[c,g][1,2]diazocine (product 14)) [ka]

[0114] After replacing the atmosphere in a two-necked recovery flask containing product 13 (2.29 g, 14.0 mmol), product 13 was dissolved in CCl4 (100 ml). NBS (N-bromosuccinimide) (2.99 g, 16.8 mmol) was then added, followed by benzoyl peroxide (17 mg, 0.07 mmol), and the mixture was heated to reflux for 24 hours. The mixture was then cooled to room temperature, and the precipitated succinimide was filtered and concentrated under reduced pressure. Product 14 (3.2 g, 80%) was obtained as a purple solid by column chromatography (ethyl acetate / hexane = 1 / 3).

[0115] The analytical results of product 14 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 3): 0.48 1H NMR (400 MHz, CDCl3 containing TMS): δ = 7.25-7.19 (2H, m), 7.10 (2H, m), 6.98 (1H, m), 5.19 (1H, m), 3.38 (1H, m), 3.12 (1H, m) ppm ESI MS (positive mode): m / z: C 14 H 11 Calculated value for BrN2: [M + Na] + 309.0003; Actual value: 308.9999

[0116] (Synthesis of (5Z,11Z)-dibenzo[c,g][1,2]diazocine (product 15)) [ka]

[0117] A two-necked eggplant flask containing product 14 (3.20 g, 11.2 mmol) was replaced with nitrogen, and product 14 was dissolved in MeOH (300 ml). KOH (3.00 g, 53.6 mmol) was added and stirred at 40°C for 27 hours. After stirring, an aqueous ammonium chloride solution was added to neutralize the mixture, and the mixture was extracted three times with ethyl acetate. The organic phase was washed once with saturated saline, and Na2SO4 was added to the separated organic phase. After filtration, the mixture was concentrated under reduced pressure. Product 15 (2.00 g, 87%) was obtained as a yellow solid by column chromatography (ethyl acetate / hexane = 1 / 4).

[0118] The analytical results of product 15 were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 3): 0.56 1H NMR (400 MHz, CDCl3 with TMS): δ = 7.26 (2H, t, J = 7.3 Hz), 7.13 (2H, t, J = 7.3 Hz), 7.05 (2H, dd, J = 7.8 Hz, J = 0.9 Hz), 7.01 (2H, dd, J = 7.8 Hz, J = 0.9 Hz), 6.72 (2H, s) ppm ESI MS (positive mode): m / z: C 14 H 10 Calculated value of N2: [M + H] + 207.0922; Actual value: 207.0922

[0119] (Synthesis of (11R,12S,Z)-11,12-dihydrodibenzo[c,g][1,2]diazocin-11,12-diol (Product 16)) [ka]

[0120] Product 15 (560 mg, 2.71 mmol) was placed in a two-necked eggplant flask and the atmosphere was replaced with nitrogen. Product 15 was dissolved in acetone / water = 1 / 1 (20 ml). N-methylmorpholine N-oxide (50%, in water, about 4.8 mol / L, 0.73 ml, 3.53 mmol) was then added, followed by microencapsulated OsO4 (content about 10%, 350 mg, 0.14 mmol), and stirred at room temperature for 24 hours. After filtration, product 16 (140 mg, 21%) was obtained as a yellow solid by column chromatography (CHCl).

[0121] The analytical results of product 16 were as follows: TLC R f (Merck 60 F254, CH2Cl2): 0.12 1H NMR (400 MHz, CDCl3 with TMS): δ = 7.53 (2H, dd, J = 7.6 Hz, J = 1.2 Hz), 7.24 (2H, t, J = 7.6 Hz), 7.18 (2H, t, J = 7.6 Hz), 6.92 (2H, dd, J = 7.6 Hz, J = 1.2 Hz), 4.97 (2H, s), 3.03 (2H, s) ppm ESI MS (positive mode): m / z: C 14 H 12 Calculated value of N2O2: [M + Na] + 263.0796; Actual value: 263.0788

[0122] ((11R,12S,Z)-11,12-bis(hexadecyloxy)-11,12-dihydrodibenzo[c,g][1,2]diazocine (Product: AzoC 16 Synthesis of [ka]

[0123] After replacing the atmosphere in a two-necked recovery flask with nitrogen, NaH (60%, dispersed in liquid paraffin, 116 mg, 2.92 mmol) was added and washed three times with dry hexane. The flask was then vacuum dried and dry THF (3 ml) was added. Then, at room temperature, a solution of 1-iodohexadecane (410 mg, 1.17 mmol) in dry THF (5 ml) was added dropwise over 15 minutes. After the addition was completed, heating was started and a solution of product 16 (70 mg, 0.29 mmol) in dry THF (10 ml) was added dropwise over 20 minutes under reflux conditions. After the addition was completed, the flask was heated under reflux for 13 hours. After cooling to room temperature, water was added to inactivate NaH, and the mixture was extracted three times with methylene chloride. Na2SO4 was added to the separated organic phase, filtered, concentrated under reduced pressure, and AzoC was isolated by silica gel column chromatography (ethyl acetate / hexane = 1 / 3). 16 (10 mg, 5%) was obtained as a yellow solid.

[0124] AzoC 16 The results of the analysis were as follows: TLC R f (Merck 60 F254, ethyl acetate / hexane = 1 / 3): 0.62 1 H NMR (400 MHz, TMS included) 1 H): δ = 7.21 (2H, d, J = 7.4 Hz), 7.12 (2H, t, J = 7.4 Hz), 6.94 (2H, t, J = 7.4 Hz), 6.86 (2H, d, J = 7.4 Hz), 4.88 (2H, s), 3.50 (4H, t, J = 6.3 Hz), 1.57 (8H, m), 1.26 (48H, s), 0.88 (6H, t, J = 6.8 Hz) ppm

[0125] [Example 3] In a Durham tube, add 2 mM phospholipid (amphiphilic molecule) (DOPC: 1,2-dioleoyl sn-glycero-3-phosphocholine) in CHCl3 / MeOH = 2 / 1 (18 μL) and 2 mM AzoC 12 -NMe3 + A MeOH solution (2 μL) of 0.18 mM ammonium nitrate was added and dried by blowing nitrogen. The mixture was then dried in a desiccator for 3 hours to produce a lipid film. MilliQ water (200 μL) was added and incubated at 37.5°C for 3 hours or more to obtain a GUV dispersion in which GUVs (Giant Unilamellar Vesicles) were dispersed in water (the concentration of amphiphilic molecules (DOPC) in water was 0.18 mM, AzoC 12 -NMe3 + concentration 0.02M).

[0126] [Example 4] A GUV dispersion was obtained in the same manner as in Example 3, except that the amphiphilic molecule was changed from DOPC to DOTAP (1,2-dioleoyl-3-trimethylammonium-propane chloride).

[0127] [Example 5] A GUV dispersion was obtained in the same manner as in Example 3, except that the amphiphilic molecule was changed from DOPC to DOPA (1,2-dioleoyl-SN-glycero-3-phosphate sodium salt).

[0128] [Example 6] The GUV dispersions obtained in Examples 3 to 5 (in water, amphiphilic molecule concentration 0.18 mM, AzoC 12 -NMe3 + The absorption spectrum was measured before and after irradiation with 400 nm light (concentration 0.02 mM) for 1 or 10 minutes. The results are shown in Figure 4. The absorption spectrum changed with an isosbestic point, indicating that AzoC 12 -NMe3 + It was suggested that the lipid bilayer that constitutes the GUV undergoes photoisomerization. Light irradiation was performed using a 300W xenon light source MAX-303 (manufactured by Asahi Spectroscopy).

[0129] [Example 7] The GUV dispersion obtained in Example 3 (in water, amphiphilic molecule (DOPC) concentration 0.18 mM, AzoC 12 -NMe3 + The behavior of the membrane deformation when the GUV dispersion (concentration 0.02 mM) was irradiated with light (wavelength 370 nm) using a 300 W xenon light source MAX-303 (manufactured by Asahi Spectroscopy) was observed with a microscope. For the microscopic observation, a 0.2 mm thick silicon film was placed on a slide glass, and a GUV dispersion (water, amphiphilic molecule (DOPC) concentration 0.18 mM, AzoC 12 -NMe3 + A 6 μL sample (concentration: 0.02 mM) was placed on the plate, and a cover glass was placed on top for observation.

[0130] After 171 seconds from the start of light irradiation, the phospholipid membrane penetrated into the inside, and after 173 seconds, another vesicle was observed to be formed inside the vesicle. In other words, the division of the vesicle into the inside was observed. The results are shown in Figure 5.

[0131] [Example 8] AzoC 12 -NMe3 + AzoC 12The same procedure as in Example 3 was carried out, except that the -NH2 was used instead, and the GUV dispersion (phospholipid (DOPC) concentration in water: 0.18 mM, AzoC 12 -NH2 concentration of 0.02M was obtained.

[0132] [Example 9] The GUV dispersion obtained in Example 8 (in water, amphiphilic molecule (DOPC) concentration 0.18 mM, AzoC 12 The behavior of the membrane deformation when light (wavelength 370 nm) was irradiated to the GUVs (-NH2 concentration 0.02 mM) using a 300 W xenon light source MAX-303 (Asahi Spectroscopy Co., Ltd.) was observed with a microscope. For the microscopic observation, a 0.2 mm thick silicon film was placed on a slide glass, and a GUV dispersion (water, amphiphilic molecule (DOPC) concentration 0.18 mM, AzoC 12 6 μL of -NH2 (concentration: 0.02 mM) was added, and a cover glass was placed on top for observation.

[0133] After 144 seconds from the start of light irradiation, the phospholipid membrane was observed to bud outward, and after 146 seconds, another vesicle was observed to form outside the vesicle. In other words, the division of the vesicle to the outside was observed. The results are shown in Figure 6.

[0134] [Example 10] In a Durham tube, 2 mM amphiphilic molecule (DOPC: 1,2-dioleoyl sn-glycero-3-phosphocholine) in CHCl3 / MeOH = 2 / 1 (19.8 μL) and 0.2 mM AzoC 16 A MeOH solution (2 μL) of 0.198 mM was added and dried by blowing nitrogen. The mixture was then dried in a desiccator for 3 hours to produce a lipid film. MilliQ water (200 μL) was added and incubated at 37.5°C for 3 hours or more to obtain a GUV dispersion in which GUVs were dispersed in water (the concentration of amphiphilic molecules (DOPC) in water was 0.198 mM, and AzoC 16 Concentration: 0.002M).

[0135] [Example 11] The GUV dispersion obtained in Example 10 (in water, amphiphilic molecule (DOPC) concentration 0.198 mM, AzoC 16The behavior of the membrane deformation when the GUV dispersion (concentration 0.002 mM) was irradiated with light (wavelength 370 nm) using a 300 W xenon light source MAX-303 (manufactured by Asahi Spectroscopy) was observed with a microscope. For the microscopic observation, a 0.2 mm thick silicon film was placed on a slide glass, and a GUV dispersion (water, amphiphilic molecule (DOPC) concentration 0.198 mM, AzoC 16 A 6 μL sample (concentration: 0.002 mM) was placed on the plate, and a cover glass was placed on top for observation.

[0136] After 109 seconds from the start of light irradiation, the phospholipid membrane was observed to bud outward, and after 120 seconds, another vesicle was observed to form outside the vesicle. In other words, the division of the vesicle to the outside was observed. The results are shown in Figure 7.

[0137] [Example 12] The GUV dispersion obtained in Example 3 (in water, amphiphilic molecule (DOPC) concentration 0.18 mM, AzoC 12 -NMe3 + The membrane deformation behavior was observed under a microscope when light (wavelength 370 nm) was irradiated using a 300 W xenon light source MAX-303 (manufactured by Asahi Spectroscopy) in the presence of anionic PS (polystyrene) beads (diameter 1 μm, 0.0026 wt% of the entire GUV dispersion) in water (concentration 0.02 mM). 12 -NMe3 + A 6 μL sample (concentration: 0.02 mM) was placed on the plate, and a cover glass was placed on top for observation.

[0138] Six hundred seconds after the start of light irradiation, it was observed that the PS beads adhering to the vesicle surface were fragmented and taken up into the interior of the vesicle due to membrane deformation. The results are shown in Figure 8.

[0139] Novel compounds having an azobenzene structure were obtained from Examples 1 and 2. It was revealed that the vesicles using the compounds obtained in Examples 1 and 2 change in structure by photoisomerization.

[0140] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

[0141] Although the present embodiment has been described in detail above, the specific configuration is not limited to this embodiment, and even if there are design changes within the scope that does not deviate from the gist of this disclosure, they are included in this disclosure.

Claims

1. A compound having an azobenzene structure represented by the following general formula (1). 【Chemistry 1】 (In the general formula (1), R １ One of the two and R ２ One side of each is independently -Z, -O-Z, -CO-O-Z, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, -NH-CO-O-Z, - O-CO-NH-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -S-Z, -CO-S-Z, -S-CO-Z, or -S-S-Z, Z's each independently represent a substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms, and at least one of the methylene structures in the substituted or unsubstituted aliphatic hydrocarbon group is -O-, -S-, -NH-, -CO-, -S-S-, or -SO ２ may be replaced by a structure selected from -, -CO-O-, -CO-NH-, and -NH-CO-O-, R １ On the other hand, R ２ The other and R ３ ~R １０ are each independently a hydrogen atom or a hydrocarbon group having 1 to 24 carbon atoms.

2. The R ３ ~R ６ At least two of R ７ ~R １０ The compound having an azobenzene structure according to claim 1 , wherein at least two of the above are hydrogen atoms.

3. 3. The compound having an azobenzene structure according to claim 1, wherein the substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms is a substituted or unsubstituted aliphatic hydrocarbon group having 5 to 35 carbon atoms.

4. 3. The compound having an azobenzene structure according to claim 1, wherein the substituted or unsubstituted aliphatic hydrocarbon group having 1 to 40 carbon atoms is a substituted or unsubstituted aliphatic hydrocarbon group having 7 to 30 carbon atoms.

5. The R １ One of the two and R ２ wherein one of the azobenzene structures is independently -Z, -O-Z, -CO-O-Z, -O-CO-Z, -CO-NH-Z, -NH-CO-Z, -NH-CO-NH-Z, -NH-CS-NH-Z, -S-Z, -CO-S-Z, -S-CO-Z, or -S-S-Z.

6. The compound having an azobenzene structure according to any one of claims 1 to 5, wherein the substituted or unsubstituted aliphatic hydrocarbon group is a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group, or a substituted or unsubstituted alkynyl group.

7. The compound having an azobenzene structure according to any one of claims 1 to 6, wherein the substituted or unsubstituted aliphatic hydrocarbon group is an unsubstituted aliphatic hydrocarbon group or an aliphatic hydrocarbon group substituted with at least one group selected from an ionic functional group and a nonionic polar functional group.

8. The R １ One of the two and R ２ wherein each of the above is independently a group selected from the following general formulas (2) to (25): 【Chemistry 2】 【Chemistry 3】 In the general formulas (2) to (25), n is each independently an integer of 5 to 15; in the general formulas (4), (6), (8), (20), (22), (24), and (25), X － is a counter anion, and in formulae (10), (12), (14), (16) and (18), Y ＋ is the counter cation.)

9. A vesicle comprising the compound having an azobenzene structure according to any one of claims 1 to 8 in a lipid bilayer.

10. The lipid bilayer contains an amphipathic molecule and a compound having an azobenzene structure, 10. The vesicle according to claim 9, comprising 0.01 to 40% by mass of the compound having an azobenzene structure relative to 100% by mass in total of the amphiphilic molecule and the compound having an azobenzene structure.

11. The vesicle of claim 10, wherein the amphiphilic molecule is a phospholipid.

12. A method for controlling a vesicle structure, comprising irradiating the vesicle according to any one of claims 9 to 11 with light to cause photoisomerization of a compound having an azobenzene structure, thereby changing the structure of the vesicle.

13. The method for controlling a vesicle structure according to claim 12, wherein the structural change of the vesicle is division into the inside of the vesicle or division into the outside of the vesicle.

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