WO2022196089A1 - Liposome complex, substance delivery system, substance delivery method, and light irradiation device - Google Patents

Abstract

The present invention provides: a liposome complex that is easy to prepare; and a technology that uses this liposome complex.　Provided is a liposome complex that includes at least: a liposome; and a photoresponsive pigment that is supported, by a non-covalent interaction, on lipids that constitute the liposome. Also provided is a material delivery system that has at least: a liposome complex that includes at least a liposome and a photoresponsive pigment that is supported, by a non-covalent interaction, on lipids that constitute the liposome; and a light irradiation device that comprises at least a light irradiation unit that irradiates light on the liposome complex.

Description

Liposome complex, substance delivery system, substance delivery method, and light irradiation device

This technology relates to a liposome complex, a substance delivery system, a substance delivery method, and a light irradiation device.

A liposome is a closed endoplasmic reticulum formed by at least one or more lipid bilayer membranes. Conventionally, liposomes encapsulating drugs and the like have been mainly used as carriers of drug delivery systems (DDS) to target tissues. Examples of the target tissue include cancer tissue.

For example, Patent Document 1 discloses a medical device that induces specific and sufficient necrosis in cancer tissue, wherein the medical device comprises a liposome membrane-constituting substance covalently bound to a light-absorbing compound, In addition, a liposome complex containing a drug inside the liposome is used.

JP 2013-230211 A

However, using covalent bonds to produce liposome complexes has the problem of requiring complex chemical synthesis to directly bond lipids and compounds. In addition, direct binding of a lipid and a compound may greatly affect the properties of the lipid, and as a result, it becomes difficult to control the properties of the liposome complex itself.

Therefore, the main purpose of this technology is to provide a liposome complex that is easy to prepare and a technology that uses the liposome complex.

That is, the present technology provides a liposome complex that includes at least a liposome and a photoresponsive dye supported by a lipid that constitutes the liposome through non-covalent interaction.

Further, in the present technology, a liposome complex including at least a liposome and a photoresponsive dye supported by a lipid constituting the liposome through non-covalent interaction, and the liposome complex is irradiated with light. Also provided is a substance delivery system comprising at least a light irradiation device comprising at least a light irradiation unit for performing.

Furthermore, in the present technology, at least a step of irradiating a liposome complex containing at least a liposome and a photoresponsive dye supported by a non-covalent interaction with a lipid constituting the liposome is performed. , also provides a method of substance delivery.

In addition, in the present technology, a light irradiation unit for irradiating a liposome complex containing at least a liposome and a photoresponsive dye supported by a non-covalent interaction with a lipid constituting the liposome, A light irradiation device is also provided, comprising at least

1 is a schematic conceptual diagram showing a first embodiment; FIG. 4A to 4D are diagrams for explaining application example 1. FIG. 4A to 4E are diagrams for explaining application example 2. FIG. 3A to 3E are diagrams for explaining application example 3. FIG. It is a schematic conceptual diagram which shows 2nd Embodiment. 4A and 4B are diagrams for explaining Experimental Example 1. FIG. FIG. 5 is a diagram showing the results of Experimental Example 1; 8A and 8B are diagrams for explaining Experimental Example 2. FIG. FIG. 10 is a diagram showing the results of Experimental Example 2; FIG. 10 is a diagram showing the results of Experimental Example 3; FIG. 10 is a diagram showing the results of Experimental Example 3; 3A to 3C are diagrams showing the results of Experimental Example 4. FIG. 7A to 7G show the results of Experimental Example 5. FIG. FIG. 10 is a diagram showing the results of Experimental Example 6;

A preferred embodiment for implementing the present technology will be described below.
It should be noted that the embodiments described below are representative embodiments of the present technology, and the scope of the present technology should not be construed narrowly. The present technology will be described in the following order.

1. First embodiment (liposome complex 10)
(1) Liposome 11
(2) Photoresponsive dye 12
(3) Inclusion 13
(4) Applications (4-1) Application example 1
(4-2) Application example 2
(4-3) Application example 3
(4-4) Application example 4
2. Second embodiment (substance delivery system 100)
3. Third embodiment (substance delivery method)
4. Fourth embodiment (light irradiation device)

1. First embodiment (liposome complex 10)

FIG. 1 is a schematic conceptual diagram showing an example of the first embodiment.
A liposome complex 10 according to this embodiment includes at least a liposome 11 and a photoresponsive dye 12 . Moreover, the inclusion 13 etc. may be included as needed.

(1) Liposome 11

The liposome 11 is a closed endoplasmic reticulum formed of at least one or more lipid bilayer membranes, and has an aqueous phase (inner aqueous phase) within the space of the closed vesicle. The liposomes 11 usually exist in a state of being dispersed in the solution outside the closed vesicles (external aqueous phase).
In the present embodiment, the liposome 11 may be a single lamella structure with a single double membrane or a multilamellar structure with multiple double membranes. If is large, it is preferably single lamellae. Moreover, the form is not specifically limited, either.

The components that make up the lipid bilayer of the liposome 11 are selected from lipids, and examples of the lipids include phospholipids, lipids that do not contain phosphoric acid, cholesterols, fatty acids, and the like, and combinations thereof may also be used.

Phospholipids include, for example, natural phospholipids such as phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidic acid, cardiolipin, sphingomyelin, egg yolk lecithin, soybean lecithin, and lysolecithin, or hydrogenated from these by a conventional method. Synthetic phospholipids such as distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleosestearoylphosphatidylcholine, eleosestearoylphosphatidylethanolamine, and eleosestearoylphosphatidylserine; .
Lipids that do not contain phosphoric acid include, for example, amino acid-type lipids, peptide lipids, glycolipids, and the like.

Cholesterols include, for example, cholesterol and phytosterols.
Examples of fatty acids include oleic acid, palmitoleic acid, linoleic acid, and fatty acid mixtures containing these unsaturated fatty acids.

In the present embodiment, the lipid is particularly preferably a lipid with phase transition among these. Here, "phase transition property" means having a phase transition point such as phase transition pressure and phase transition temperature. In addition, "phase transition" means a transition that occurs between two states, a liquid crystal phase and a gel phase, in a lipid bilayer.

(2) Photoresponsive dye 12

The photoresponsive dye 12 is supported by the lipids constituting the liposome 11 through non-covalent interactions. Here, the term "photoresponsive" means that the property changes reversibly or irreversibly when irradiated with light. Specifically, for example, when the photoresponsive dye 12 is irradiated with light, an absorption spectrum is observed. In this embodiment, the absorption wavelength of the photoresponsive dye 12 is not particularly limited, but is preferably 200 nm to 2000 nm, more preferably 300 nm to 1000 nm.

In addition, the term “carrying” refers to a form in which the entire photoresponsive dye 12 is contained in the membrane of the lipid constituting the liposome 11, a form in which only a part thereof is contained in the membrane, or a form in which the photoresponsive dye 12 is contained in the membrane. It means taking a form that exists outside the membrane in a state where the whole is attached to the membrane.

In the present embodiment, the photoresponsive dye 12 is supported by the lipid through non-covalent interaction, which facilitates the preparation of the liposome complex 10 according to the present embodiment. If the photoresponsive dye is covalently bound to the lipid, a very complicated chemical synthesis is required for the preparation of the liposome complex. On the other hand, when the photoresponsive dye 12 is supported on the lipid using a non-covalent bond, the liposome complex 10 can be prepared simply by performing operations such as mixing in an appropriate solvent and drying. It is possible to

It is also easy to change the combination of the photoresponsive dye 12 and the lipid as appropriate according to the application of the liposome complex 10 and the like. As described above, the preparation of the liposome complex 10 is easier than the case where the photoresponsive dye is covalently bound to the lipid. Therefore, it is possible to prepare the liposome complex 10 having various properties. It becomes possible. Specifically, for example, by changing the photoresponsive dye 12, it is possible to make the liposome complex 10 multicolor, or to change the phase transition temperature by using a combination of lipids having different phase transition temperatures. can.

Furthermore, covalent bonding of photoresponsive dyes to lipids may have a significant impact on the lipid aggregation properties. Therefore, it is necessary to design the photoresponsive dye-lipid conjugate each time, and chemically synthesize it each time, which makes it difficult to control the properties of the liposome complex. On the other hand, when the photoresponsive dye 12 is supported on the lipid using a non-covalent bond, the characteristics of the photoresponsive dye 12 and the lipid constituting the liposome 11 are maintained. Therefore, it is easy to predict the properties of the liposome complex 10 itself, and as a result, it is easy to control the properties.

Examples of non-covalent interactions include any one or more interactions selected from the group consisting of electrostatic interactions, hydrophobic interactions, van der Waals interactions, and π interactions.

Electrostatic interactions include, for example, interactions between ions, hydrogen bonds, interactions between halogens, or combinations thereof.
Van der Waals interactions include, for example, dipole-dipole interaction, dipole-induced dipole interaction, induced dipole-induced dipole interaction (London dispersion force) and the like.
Examples of π interaction include π-π interaction, cation-π/anion-π interaction, metal-π interaction, polar-π interaction, π donor-acceptor interaction, CH-π interaction, and the like. mentioned.

In the present embodiment, the non-covalent action is particularly preferably hydrophobic interaction or a combination of hydrophobic interaction and other non-covalent action.

As the photoresponsive dye 12, for example, the following molecules can be used singly or in combination. When used in combination, a substituent or the like described later may be used.

Specifically, for example, polymethines such as cyanine, squalane, croconine, merocyanine, curcumin; phthalocyanine, naphthalocyanine, subphthalocyanine; porphyrin, chlorin, chlorophyll, azaporphyrin, corole, cyanocobalamin, oligophyrin; oligophenol, polyphenol, melanin , quinone; diimonium; metal complexes such as dithiolate complexes, rare earth complexes and transition metal complexes; dipyrromethene such as BODIPY; spiro compounds such as spiropyran and spiroperimidine; azo compounds such as azobenzene; , naphthalene, anthracene; perylene diimide, naphthalene diimide; indigo, thioindigo; acridine, quinacridone; phenothiazine; structures in which heteroatoms are substituted at each position of the structure; structures in which a plurality of the above conjugated structures are connected or condensed;

A portion of the photoresponsive dye 12 may be substituted with one or more substituents. In this case, the substituent preferably has a high affinity with the lipid that constitutes the liposome. The substituent can be appropriately selected from, for example, the following substituents, and a single or multiple types of substituents can be used in combination. Moreover, in this embodiment, the photoresponsive dyes 12 may be connected to each other via a substituent.

Specifically, for example, linear, branched or cyclic alkyl groups; halogenated alkyl groups such as partial fluoroalkyl groups and perfluoroalkyl groups; unsaturated carbonized groups such as phenyl groups, ethynyl groups, vinyl groups and pyrene groups; Hydrogen groups, or linear or condensed conjugated molecules connecting them; Aromatic rings containing heteroatoms such as thiophene, pyrrole, furan, pyridine, aniline groups, etc., or their condensed rings; Halogen-containing organic compounds, halides; silyl compounds such as silylalkyl groups, silylalkoxy groups, and arylsilyl groups; Sulfur compounds such as carbonyl groups; nitrogen compounds such as amino groups, alkylamino groups, arylamino groups, amide groups, ammonium groups, nitro groups, cyano groups; hydroxy groups, alkoxy groups, phenoxy groups, aryloxy groups, acyl groups, Oxygen compounds such as acylamino group, acyloxy group, carboxy group, carboxamide group, carboalkoxy group, formyl group, thioacyl group; vinyl group, allyl group, (meth)acrylic group, glycidyl group, aziridine ring, isocyanate group, conjugated diene , acid anhydride, acid chloride, carbonyl group, hydroxyl group, amide group, amino group, chloromethyl group, ester group, formyl group, nitrile group, nitro group, carbodiimide group, oxazoline group and other polymerizable functional groups; boron compound ; a chalcogenide compound; and the like, and a combination thereof may also be used. Also, metals such as alkali metals, alkaline earth metals and transition metals may be coordinated through the above substituents.

The liposome complex 10 according to the present embodiment releases an inclusion 13, which will be described later, when irradiated with light. Here, "release" is a broad concept including stepwise sustained release, large-scale release at one time, and the like. Moreover, in this embodiment, it is possible to control the release amount of the inclusion 13, the release rate, and the like.

Specifically, for example, by arranging the liposome complex 10 in the vicinity of the target and irradiating it with light, the inclusions 13 can be released and act on the target. Here, the “object” is not particularly limited, but includes, for example, cells, biological tissue, etc., and the surface of materials such as paper, resin, metal, etc., and the inclusions 13 are directly acted inside and outside, or liposome complexes are used. Anything that the body 10 can react to as the external environment can be targeted.

More specifically, for example, an object is placed in an optical system such as a microscope, the dispersion liquid of the liposome complex 10 containing the inclusions 13 is brought into contact, any liposome complex 10 is irradiated with light, and the inclusions 13 are exposed. is released. A more specific example will be described in detail in "(4) Application" described later.

In this embodiment, the size of the liposome complex 10 can be, for example, 1 nm or more and 1 mm or less, preferably 20 nm or more and 100 μm or less. By setting the size of the liposome complex 10 to 1 nm or more and 1 mm or less, the liposome complex 10 can be directly irradiated with light.

When the liposome complex 10 is irradiated with light, the energy of the light is preferably less than the energy at which the temperature changes in the vicinity of the liposome complex 10 irradiated with the light. When the photoresponsive dye is covalently bound to the lipid, the entire surroundings are heated when the encapsulation is released, or a large amount of heat is generated by the stimulus for release. It affects the surroundings (for example, it damages surrounding normal cells, etc.), and it is difficult to obtain the action of the inclusion alone. On the other hand, in the present embodiment, the photoresponsive dye 12 is carried by non-covalent interaction with the lipid that constitutes the liposome 11, and the affinity between the photoresponsive dye 12 and the lipid is high. Even relatively small energy can cause the inclusion 13 to be released. In addition, by setting the energy of the light to be less than the energy at which the temperature changes in the vicinity of the liposome complex 10 irradiated with the light, the effects of temperature rise on the surroundings are suppressed, and the effect is non-invasive. , it can be expected that only the action of the inclusion 13 can be obtained while reducing undesirable stimulation as much as possible.

In this embodiment, it is preferable that the molar ratio between the lipid constituting the liposome 11 and the photoresponsive dye 12 is 10 ⁸ :1 or more. This allows the photoresponsive dye 12 to efficiently stimulate the lipid membrane.

Further, in the present embodiment, the photoresponsive dye 12 is preferably a photothermal conversion dye. This is because heat stimulation is particularly preferable as the noninvasive stimulation means in this embodiment.

Furthermore, in this embodiment, the photoresponsive dye 12 preferably has photostability. Here, the term "light stability" means durability when irradiated with light. A dye having photostability is, for example, a dye that has the property of being difficult to decompose when the dye is irradiated with light. More specifically, for example, when a solution containing the dye is continuously irradiated with light, the temperature rise curve converges and the temperature does not drop after a certain time. The above time varies depending on the pigment skeleton, pigment concentration, oxygen saturation, power of light irradiation, and the like.

Examples of the photoresponsive dye 12 having photostability include, among the specific examples of the photoresponsive dye 12 described above, an aromatic ring, a condensed ring conjugate system, or a condensed or condensed ring thereof. . In addition, even in a linear conjugated molecule having a low aromatic structure such as ethylene or acetylene, it is possible to improve the photostability by introducing a substituent for photostabilization. Specifically, for example, in the case of a cyanine-based polymer, photostability can be imparted by introducing thiophenol or the like. As the substituent for photostabilization, the above-described substituents and the like can be used, excluding substituents with high eliminability such as halogen. Photostability can also be imparted to the photoresponsive dye 12 by binding a radical scavenger to it or arranging it in the vicinity.

(3) Inclusion 13

In this embodiment, the inclusion 13 may be included as necessary.

The inclusion 13 is not particularly limited as long as it can be included in the liposome 11 described above. Here, the term “encapsulated” means that the encapsulated matter 13 is contained in the inner aqueous phase and the membrane itself of the liposome 11 . Specifically, for example, there are a form in which the inclusion 13 is enclosed in a closed space formed of a membrane, a form in which the inclusion is enclosed in the membrane itself, and the like, and a combination thereof is also possible.

Specifically, for example, acetylcholine, lipids, signaling substances, ATP, mRNA, calcium ions, metal ions, other various ions, drugs, physiologically active substances, amino acids, peptides, proteins, nucleic acids, sugars, reducing agents, oxidation agents, acidic substances, alkaline substances, and the like. The inclusion 13 may be either water-insoluble or water-soluble, but among these, water-soluble is particularly preferred.

(4) Applications

Hereinafter, applications of the liposome complex 10 according to the present embodiment will be described with examples.
Application examples 1 to 4 described below show typical application examples of the liposome complex 10 according to the present embodiment, and thus the range of application of the liposome complex 10 according to the present embodiment. cannot be interpreted narrowly.

(4-1) Application example 1
FIG. 2 shows an example of a substance transport process to living tissue, cells, and the like. As shown in FIGS. 2A and 2B, the subject is brought into contact with the dispersion liquid of the liposome complex 10 encapsulating the drug or the like, and as shown in FIG. By releasing the included drug or the like, the drug or the like reaches a desired position, range, or the like, and as shown in FIG. At this time, the liposome complex 10 according to the present embodiment can be manipulated non-invasively, since the liposome complex 10 according to the present embodiment causes less stimulation such as heat to be applied to the surroundings. Therefore, it can be used for tissues such as nervous tissue, brain tissue, etc., which are sensitive to heat, and delicate tissues, such as skin.

In addition, it is also possible to target molecules such as proteins in the substance transport process mentioned above. For example, by appropriately selecting lipids constituting the liposome or functionalizing the lipid surface, it is possible to target a specific molecule and allow the inclusion 13 to act on the molecule. As a result, the inclusion 13 can act in the vicinity of the target molecule at a high concentration, so that the reaction efficiency, speed, etc. between the target molecule and the inclusion 13 are increased. Therefore, since the reaction efficiency is high, the amount of the inclusion 13 can be reduced, and as a result, for example, the dose to be administered into the body can be suppressed.

(4-2) Application example 2

Fig. 3 shows an example of a material deposition process such as plating. As shown in FIGS. 3A and 3B, the object is allowed to carry a catalyst or the like (for example, the catalyst or the like is fixed to a base material), and as shown in FIG. 10 are brought into contact with each other, and as shown in FIG. 3D, a desired position, range, or the like is irradiated with light to gradually release the contained metal ions or the like. After that, as shown in FIG. 3E, the metal or the like can be gradually precipitated around the areas where the sustained release has occurred. At this time, since the liposome complex 10 according to the present embodiment does not generate thermal convection, the deposition rate can be controlled with high accuracy, and a smooth surface can be obtained.

(4-3) Application example 3

An example of the crystallization process is shown in Fig. 4. As shown in FIGS. 4A and 4B, the substrate is brought into contact with the dispersion liquid of the liposome complex 10 encapsulating organic monomers, inorganic monomers, etc., which are the source of crystals, and as shown in FIG. The material or the entire system is heated or cooled to prepare crystal growth conditions. Next, as shown in FIG. 4D, a desired position, range, or the like is irradiated with light to gradually release the encapsulated organic monomer, inorganic monomer, or the like. After that, as shown in FIG. 4E, crystals can be slowly grown on the base material or in the system, centering on the slowly released portion. At this time, since thermal convection does not occur in the liposome complex 10 according to the present embodiment, the deposition rate can be controlled with high accuracy, and crystals with a high degree of crystallinity can be obtained.

(4-4) Application example 4

The liposome complex 10 according to the present embodiment can be applied even if its shape is not spherical as long as it has a membrane composed of lipids. For example, if a lipid membrane placed in a microchannel or the like is used as the liposome complex 10, the channel can be gated or closed by irradiating a desired position, range, or the like with light.

2. Second embodiment (substance delivery system 100)

FIG. 5 is a conceptual schematic diagram showing an example of the second embodiment.
A substance delivery system 100 according to this embodiment includes at least the liposome complex 10 and the light irradiation device 101 described above. In addition, it may have other devices or the like as necessary. Since the liposome complex 10 is the same as described above, the explanation is omitted here.

(1) Light irradiation device 101

The light irradiation device 101 irradiates the liposome complex 10 with light. The light irradiation device 101 includes, for example, a light irradiation unit 102 that irradiates laser light, an irradiation condition setting unit 103 that can change the frequency, irradiation intensity, pulse width, etc. of the laser light to be irradiated, movement of the irradiation area, irradiation area etc., a light-receiving unit 105 that receives light emitted from a target in response to the laser light emitted from the light-irradiating unit 102, and an analyzing unit 106 that analyzes the light obtained from the light-receiving unit 105. And prepare.

In this embodiment, the light irradiation unit 102 can also irradiate the liposome complex 10 with light of two or more different wavelengths. Also, an external analysis device or the like may be used instead of the analysis unit 106 . Specifically, for example, it may be implemented in a personal computer or CPU, and stored as a program in a hardware resource including a recording medium (eg, nonvolatile memory (USB memory), HDD, CD, etc.), and stored in a personal computer. or by a CPU. Also, an external analysis device or the like may be connected to each part of the light irradiation device 101 via a network.

3. Third embodiment (substance delivery method)

The substance delivery method according to this embodiment performs at least the step of irradiating the liposome complex 10 with light. Moreover, you may have another process as needed. Since the liposome complex 10 is the same as described above, the explanation is omitted here. Further, since the method performed in the above steps is the same as the method performed by the light irradiation device 101 described above, the description is omitted here.

4. Fourth embodiment (light irradiation device)

The light irradiation device according to this embodiment includes at least a light irradiation unit that irradiates the liposome complex 10 described above with light. In addition, it may have other parts or the like as necessary. Since the liposome complex 10 is the same as described above, the explanation is omitted here. Further, since the processing performed in the light irradiation unit is the same as the processing performed in the light irradiation device 101 described above, the description thereof is omitted here.

Hereinafter, the present technology will be described in further detail based on examples.
It should be noted that the embodiments described below are examples of representative embodiments of the present technology, and the scope of the present technology should not be interpreted narrowly.

<Preparation example>

1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), N-(3-carboxy-1-oxopropyl)-, 1,5-dihexadecyl ester (SA), 1,2-Distaroyl-sn-glycero- 3-phosphoethanolamine-N-[monomethoxy poly(ethylene glycol) (2000)] (PEG-DSPE) was mixed at a molar ratio of 9:1:0.06, and 0.05, 0.5 mM was added to 5 mg of the resulting mixed lipid. 0.5 mL of dye chloroform solution was added and dissolved, and chloroform was distilled off in a glass vial. An aqueous solution in which a drug, a fluorescent dye, a pH adjuster, an ion concentration adjuster, etc. were dissolved was added to the obtained lipid-pigment film to hydrate it, thereby obtaining a suspension. The resulting suspension was fractionated by gel chromatography to isolate only the liposome component to obtain a liposome complex.

In addition, in this preparation example, compounds represented by the following chemical formulas (1) to (6) were used as the dyes.

<Experimental example 1>

A compound represented by the chemical formula (4) was selected as the dye, and a liposome complex was prepared in the same manner as in the above preparation example. Next, as shown in FIGS. 6A and 6B, the calcein-encapsulating liposome complex is placed in a temperature probe solution whose fluorescence intensity varies with temperature, and laser light is irradiated to generate The ambient temperature change due to heat (circled points 1, 2 and 3 in FIG. 6B) was estimated. FIG. 7 shows changes in the fluorescence intensity ratio derived from the temperature probe at each point during laser light irradiation. As shown in FIG. 7, there was no fluorescence intensity change at any point, and no temperature change due to heat emitted from the liposome complex was observed.

<Experimental example 2>

A compound represented by the chemical formula (4) was selected as the dye, and a liposome complex was prepared in the same manner as in the above preparation example. Next, as shown in FIG. 8, a temperature probe was directly bound to the liposome complex, calcein was incorporated into the liposome complex, and temperature changes in the vicinity of the liposome complex during laser light irradiation were observed. FIG. 9 shows changes in the fluorescence intensity ratio during laser light irradiation. α in FIG. 9 indicates changes in the fluorescence intensity ratio derived from the temperature probe, and β in FIG. 9 indicates changes in the fluorescence intensity ratio derived from calcein. As shown in FIG. 9, no change in temperature was observed in the vicinity of the liposome complex, while the fluorescence intensity in the center changed. Therefore, it was found that inclusions can be released without changing the temperature in the vicinity of the liposome complex.

<Experimental example 3>

A compound represented by the chemical formula (4) was selected as the dye, and a liposome complex was prepared in the same manner as in the above preparation example. Next, the calcein-encapsulated liposome complex was irradiated with laser light under different temperature conditions of 37°C or 25°C. FIG. 10 shows changes in fluorescence intensity ratio derived from calcein during laser light irradiation under a temperature condition of 37°C. Further, FIG. 11 shows changes in fluorescence intensity ratio derived from calcein during laser light irradiation under a temperature condition of 25°C. As shown in FIGS. 10 and 11, it was found that since the stimulation by laser light irradiation was effectively transmitted to the lipids constituting the liposomes, it was possible to make the inclusions leak out with less energy than in the past. rice field. It was also found that the gradual release of the encapsulation is possible without collapsing the liposome complex. Furthermore, it was also found that the inclusions can be released even at low temperatures that are far from the phase transition temperature of the lipids that constitute the liposomes. Therefore, the liposome complex can be used not only in vivo but also in industrial applications.

<Experimental example 4>

In Experimental Example 4, each compound represented by Chemical Formula (1), Chemical Formula (4), and Chemical Formula (5) was selected as the dye, and each liposome complex was prepared in the same manner as in the above Preparation Examples. . Next, the calcein-encapsulated liposome complex was irradiated with a laser beam at a temperature of 25°C. FIG. 12A shows the change in fluorescence intensity ratio derived from calcein upon laser light irradiation when the compound represented by the chemical formula (4) is selected, and FIG. 12B shows the compound represented by the chemical formula (1). FIG. 12C shows the change in the fluorescence intensity ratio derived from calcein during laser light irradiation when the compound represented by the chemical formula (5) was selected. shows a change in As shown in FIG. 12, it was found that the dye composed of the compound represented by the chemical formula (1) did not release inclusions even when irradiated with laser light, and did not exhibit photoresponsiveness.

<Experimental example 5>

In Experimental Example 5, each compound represented by Chemical Formulas (1) to (6) was selected as the dye, and each liposome complex was prepared in the same manner as in the above Preparation Examples. Next, the absorption spectrum of the suspension of each liposome complex was measured. FIG. 13 shows the result of measuring the absorption spectrum of the suspension of each liposome complex. A in FIG. 13 is a negative control, B in FIG. 13 is when the compound represented by the chemical formula (1) is selected, and C in FIG. 13 is when the compound represented by the chemical formula (2) is selected. D is when the compound represented by the chemical formula (3) is selected, E in FIG. 13 is when the compound represented by the chemical formula (4) is selected, and F in FIG. 13 is when the compound represented by the chemical formula (5) is selected , and G in FIG. 13 show the case where the compound represented by the chemical formula (6) is selected. As shown in FIG. 13, a broad absorption spectrum derived from the absorption of the dye was observed except for the dye made of the compound represented by the chemical formula (1). not observed.

This is related to the affinity between each dye and the lipid that constitutes the liposome, and the dye enters the lipid in a system with high affinity, that is, the interaction between the two is strong, or the dye is adsorbed in the vicinity. It is conceivable that it will become a starting point for transmitting stimulation by laser light. Therefore, the dye composed of the compound represented by the chemical formula (1) has low interaction with lipids and is not incorporated into lipids, resulting in a liposome complex composed of the dye and lipids having low photoresponsiveness. it is conceivable that.

<Experimental example 6>

In this Experimental Example 6, each compound represented by the chemical formulas (2) to (6), excluding the compound represented by the chemical formula (1), was selected as the dye, and each dye was dissolved in an organic solvent and allowed to stand at room temperature. While irradiating the laser light, the temperature change of each solution was observed over time. FIG. 14 shows the results of measuring the temperature of each solution over time. As shown in FIG. 14, it can be seen that the temperature of each dye increases with the irradiation time, and the light is converted into heat. going from rising to falling. This is due to the fact that the dye made of the compound represented by the chemical formula (5) has low photostability, and the dye decomposes as it is irradiated with laser light, resulting in a decrease in absorbance. It captures the phenomenon of being unable to convert and returning to room temperature. That is, since the dye composed of the compound represented by the chemical formula (5) has low photostability, it cannot be decomposed in lipids to convert light into heat. As shown in Example 5, the photoresponsivity was found to be low.

Note that the following configuration can be adopted in the present technology.
[1]
a liposome;
a photoresponsive dye supported by a non-covalent interaction on the lipid constituting the liposome;
A liposome complex comprising at least
[2]
[1], wherein the non-covalent interactions are any one or more interactions selected from the group consisting of electrostatic interactions, hydrophobic interactions, van der Waals interactions, and π interactions; liposome complex.
[3]
The liposome complex according to [1] or [2], wherein the photoresponsive dye is partially substituted with one or more substituents.
[4]
The liposome complex according to any one of [1] to [3], which releases the entrapped substance upon irradiation with light.
[5]
The liposome complex according to [4], wherein the liposome complex has a size of 1 nm or more and 1 mm or less.
[6]
The liposome complex according to [4] or [5], wherein the energy of the light is less than the energy at which the temperature changes in the vicinity of the liposome complex irradiated with the light.
[7]
The liposome complex according to any one of [1] to [6], wherein the molar ratio between the lipid and the photoresponsive dye is 10 ⁸ :1 or more.
[8]
The liposome complex according to any one of [1] to [7], wherein the photoresponsive dye is a photothermal conversion dye.
[9]
The liposome complex according to any one of [1] to [8], wherein the photoresponsive dye has photostability.
[10]
a liposome complex comprising at least a liposome and a photoresponsive dye supported by a lipid constituting the liposome through non-covalent interaction;
A substance delivery system comprising at least a light irradiation device including at least a light irradiation unit for irradiating the liposome complex with light.
[11]
a step of irradiating a liposome complex containing at least a liposome and a photoresponsive dye supported by a non-covalent interaction with a lipid constituting the liposome with light;
A method of substance delivery comprising:
[12]
a light irradiation unit for irradiating a liposome complex containing at least a liposome and a photoresponsive dye supported by a non-covalent interaction with a lipid constituting the liposome;
A light irradiation device comprising at least

10: Liposome Complex 11: Liposome 12: Photoresponsive Dye 13: Inclusion 100: Substance Delivery System 101: Light Irradiation Device 102: Light Irradiation Unit 103: Irradiation Condition Setting Unit 104: Adjusting Means 105: Light Receiving Unit 106: Analysis Department

Claims (12)

1. a liposome;
   a photoresponsive dye supported by a non-covalent interaction on the lipid constituting the liposome;
   A liposome complex comprising at least
2. 2. The non-covalent interaction according to claim 1, wherein the interaction is any one or more selected from the group consisting of electrostatic interaction, hydrophobic interaction, van der Waals interaction, and π interaction. liposome complex.
3. The liposome complex according to claim 1, wherein the photoresponsive dye is partially substituted with one or more substituents.
4. The liposome complex according to claim 1, which releases the entrapped material when irradiated with light.
5. The liposome complex according to claim 4, wherein the size of the liposome complex is 1 nm or more and 1 mm or less.
6. The liposome complex according to claim 4, wherein the energy of the light is less than the energy at which the temperature changes in the vicinity of the liposome complex irradiated with the light.
7. The liposome complex according to claim 1, wherein the molar ratio of said lipid and said photoresponsive dye is ¹⁰⁸ :1 or more.
8. The liposome complex according to claim 1, wherein the photoresponsive dye is a photothermal conversion dye.
9. The liposome complex according to claim 1, wherein the photoresponsive dye has photostability.
10. a liposome complex comprising at least a liposome and a photoresponsive dye supported by a lipid constituting the liposome through non-covalent interaction;
    A substance delivery system comprising at least a light irradiation device including at least a light irradiation unit for irradiating the liposome complex with light.
11. a step of irradiating a liposome complex containing at least a liposome and a photoresponsive dye supported by a non-covalent interaction with a lipid constituting the liposome with light;
    A method of substance delivery comprising:
12. a light irradiation unit for irradiating a liposome complex containing at least a liposome and a photoresponsive dye supported by a non-covalent interaction with a lipid constituting the liposome;
    A light irradiation device comprising at least
    
    

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