CN116947956B - Diselenide with targeting and responsiveness, and preparation method and application thereof - Google Patents

Diselenide with targeting and responsiveness, and preparation method and application thereof Download PDF

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CN116947956B
CN116947956B CN202310937951.0A CN202310937951A CN116947956B CN 116947956 B CN116947956 B CN 116947956B CN 202310937951 A CN202310937951 A CN 202310937951A CN 116947956 B CN116947956 B CN 116947956B
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diselenide
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CN116947956A (en
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景苏
吴春燕
张磊
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Nanjing Taiaode Biotechnology Co ltd
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Nanjing Tech University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J51/00—Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/02—Acyclic radicals, not substituted by cyclic structures
    • C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical

Abstract

The invention relates to the field of self-assembly research, and relates to diselenide with targeting and responsiveness, a preparation method and application thereof. And carrying out dynamic exchange photoreaction on galactose diselenide and cholesterol diselenide in a photoreactor to obtain the compound containing the amphipathic diselenide. The invention uses alpha-D-pentaacetyl galactose, 11-bromoundecanol, 11-bromoundecanoic acid, cholesterol and selenium powder as raw materials to respectively synthesize small molecule galactose diselenide and cholesterol diselenide containing diselenide bond and carbon chain; the alpha-galactoside terminal configuration in the synthesis process of galactose diselenide can be turned over to be changed into beta-galactoside terminal, so that the targeting capability of tumor cells is realized; cholesterol diselenide not only serves as a hydrophobic end and provides hydrophobic acting force for the self-assembly process, but also can increase the biocompatibility of the carrier in organisms.

Description

Diselenide with targeting and responsiveness, and preparation method and application thereof
Technical Field
The invention relates to the field of self-assembly research, and relates to diselenide with targeting and responsiveness, a preparation method and application thereof.
Background
The cell surface glycoconjugates include glycoproteins and glycolipids, wherein a protein or lipid molecule is connected with a glycan by a glycosidic bond, and the glycan is branched into a sugar chain by a plurality of monosaccharides including beta-galactose by the glycosidic bond to carry complex information, and all substances containing the glycosidic bond are collectively called glycoside. Beta-galactoside and Galectin are also found in the sugar complexes on the surfaces of tumor cells and immune cells, and can be identified and specifically combined with the beta-galactoside in the sugar complexes, and the interaction between the beta-galactoside and the Galectin not only exists in the pathological process of tumor cell development and the physiological process of differentiation activation of immune cells, but also participates in the immune identification and response of the immune cells to the tumor cells, so that Galectin is increasingly focused in the tumor immunity field.
Selenium (Se) containing compounds are widely used as antioxidants in pharmaceutical chemistry for glutathione peroxidase (GPx) activity, where dienamides are a promising double redox reaction candidate due to their good activity in the presence of oxidizing or reducing agents. In general, selenium-selenium bond generates selenonic acid in the presence of oxidant and is reduced to selenol in reducing environment, and has drug response release in tumor microenvironment. The diselenide bond serves as a dynamic covalent bond, and the dynamic exchange reaction is performed under mild conditions, which is caused by irradiation with visible light and stops in the dark. This is because diselenide bonds have a lower bond energy than disulfide bonds (diselenide bonds: 172kJ/mol; disulfide bonds: 240 kJ/mol). And compared with visible light, the ultraviolet irradiation can shorten the time of dynamic exchange reaction. Dynamic diselenide linkages and their metathesis have potential uses in the modification of biological molecules, such as certain diselenide linkage-containing proteins. In addition, the double decomposition of diselenide can synthesize different diselenide linked amphiphilic polymers and manufacture visible light induced self-repairing materials containing diselenide bonds. As selenium related chemistry evolves, more applications of diselenide metathesis will be found.
Amphiphilic molecules can self-assemble in water to form a variety of ordered aggregates (most typically micelles and vesicles, for example) that are in a phase (including single or multiple mixed phases). Amphiphilic molecules generally comprise a polar region and a non-polar region, which when dispersed in water can form aggregates of a variety of morphologies, including monolayer films, bilayer or multilayer films, spherical micelles, rod-like micelles, emulsions, discoids, vesicles, cubic structures, nanopatterned structures, and the like. The polymer has a special aggregation form, so that the polymer has wide application in the fields of biological medicine, nanotechnology and the like. However, when the amphiphilic molecules are self-assembled into aggregates to be used as drug carriers, the problems of no targeting to tumor cells, no responsiveness in tumor microenvironment, poor biocompatibility and the like still exist, and research on molecular self-assembly and intelligent molecular self-assembly is a hot spot for future research.
Disclosure of Invention
The invention aims to solve the technical problems that a drug carrier formed by self-assembly of amphiphilic micromolecules in the prior art lacks targeting to tumor cells, lacks responsiveness in tumor microenvironment, cannot release drugs as required, has poor biocompatibility and the like, and provides an amphiphilic diselenide compound with good targeting, responsiveness and biocompatibility, namely galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 Chol), which self-assembles in aqueous solution to form micelles.
The invention is characterized in that: galactose diselenide ((Gal-C) 11 -Se) 2 ) And cholesterol diselenide ((Chol-C) 10 -Se) 2 ) The Se-Se bond is dynamic chemical bond, and can be dynamically exchanged under ultraviolet irradiation to obtain amphiphilic diselenide compound (Gal-C) with hydrophilic galactose at one end and hydrophobic cholesterol at the other end 11 -Se-Se-C 10 Chol), the amphiphilic diselenide compound self-assembles into micelles with drug carrying capacity in aqueous solution.
In order to solve the technical problems, the invention adopts the following technical scheme:
the invention discloses an amphipathic diselenide compound, which has a structural formula shown in a formula VI:
the invention further discloses a preparation method of the amphipathic diselenide compound, which comprises the following steps: dissolving a compound III in a first solvent to obtain a first mixed solution; dissolving the compound V in a second solvent to obtain a second mixed solution; uniformly mixing the first mixed solution and the second mixed solution, then placing the mixed solution into a photoreactor, performing dynamic exchange photoreaction to obtain a reaction solution containing an amphipathic diselenide compound, and performing aftertreatment on the reaction solution to obtain the amphipathic diselenide compound, namely a compound VI;
In some embodiments, the method of preparing compound iii comprises the steps of:
(1) The alpha-D-pentaacetyl galactose and 11-bromo-1-undecanol are subjected to etherification reaction under the action of a first catalyst, so that a compound I is prepared;
(2) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding the compound I obtained in the step (1) and a second catalyst into a reaction solution containing sodium diselenide, and performing a first coupling reaction to obtain a compound II;
(3) Performing hydrolysis reaction on the compound II obtained in the step (2) under the action of alkali to obtain galactose diselenide, namely a compound III;
in some embodiments, in step (1), the first catalyst is anhydrous zinc chloride; the molar ratio of the alpha-D-pentaacetyl galactose to the 11-bromo-1-undecanol is 1:1.33:1.33; the etherification reaction is carried out at the reaction temperature of 60-80 ℃ for 6-9 h.
In some embodiments, preferably, in step (1), the first catalyst is anhydrous zinc chloride; the molar ratio of the alpha-D-pentaacetyl galactose to the 11-bromo-1-undecanol is 1:1.33:1.33; the etherification reaction is carried out at the reaction temperature of 80 ℃ for 9 hours.
Wherein, the solvent used in the etherification reaction is anhydrous toluene; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
In some embodiments, in step (2), the second catalyst is tetrabutylammonium bromide; the molar ratio of the selenium powder to sodium borohydride to the compound I to the second catalyst is 1-1.5: 1 to 1.5:1:0.3; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 35-70 min; the first coupling reaction is carried out at room temperature for 8-12 h.
In some embodiments, preferably, in step (2), the second catalyst is tetrabutylammonium bromide; the molar ratio of the selenium powder to sodium borohydride to the compound I to the second catalyst is 1.5:1.5:1:0.3; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 65min; the first coupling reaction is carried out at room temperature for 12 hours.
Wherein the solvent used in the reduction reaction is ultrapure water; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the reduction reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
Wherein the solvent used in the first coupling reaction is tetrahydrofuran; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the first coupling reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
In some embodiments, in step (3), the base is sodium methoxide; the molar ratio of the compound II to the alkali is 1:1.8; the hydrolysis reaction is carried out at room temperature for 1-2 h.
In some embodiments, preferably, in step (3), the base is sodium methoxide; the molar ratio of the compound II to the alkali is 1:1.8; the hydrolysis reaction is carried out at room temperature for 1h.
Wherein, the solvent used in the hydrolysis reaction is a mixed solution of methanol and dichloromethane in any proportion, and the volume ratio of methanol to dichloromethane is preferably 1: 2; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
In some embodiments, the method of preparing compound v comprises the steps of:
(i) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding 11-bromoundecanoic acid into a reaction solution containing sodium diselenide, and performing a second coupling reaction to prepare a compound IV;
(ii) The compound IV obtained in the step (i) and cholesterol are subjected to esterification reaction under the action of a dehydrating agent and a third catalyst to obtain cholesterol diselenide, namely a compound V;
in some embodiments, in step (i), the molar ratio of selenium powder to sodium borohydride, 11-bromoundecanoic acid is 1:1:1, a step of; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 35-70 min; the second coupling reaction is carried out at room temperature for 8-12 h.
In some embodiments, preferably, in step (i), the molar ratio of selenium powder to sodium borohydride, 11-bromoundecanoic acid is 1:1:1, a step of; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 65min; and the second coupling reaction is carried out at room temperature for 12 hours.
Wherein the solvent used in the reduction reaction is ultrapure water; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the reduction reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
Wherein the solvent used in the second coupling reaction is tetrahydrofuran; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the second coupling reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
In some embodiments, in step (ii), the dehydrating agent is N, N-dicyclohexylcarbodiimide; the third catalyst is 4-dimethylaminopyridine; the molar ratio of the compound IV to cholesterol, the dehydrating agent and the third catalyst is 1:2:2.1:0.03; the esterification reaction is carried out at room temperature for 10-15 h.
In some embodiments, preferably, in step (ii), the dehydrating agent is N, N-dicyclohexylcarbodiimide; the third catalyst is 4-dimethylaminopyridine; the molar ratio of the compound IV to cholesterol, the dehydrating agent and the third catalyst is 1:2:2.1:0.03; the esterification reaction is carried out at room temperature for 12 hours.
Wherein the solvent used in the esterification reaction is ultra-dry dichloromethane; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
In some embodiments, in the method of preparing an amphiphilic diselenide compound, the first solvent is deuterated methanol or methanol; the second solvent is deuterated chloroform or dichloromethane; the mass-volume ratio of the total mass of the compound III and the compound V to the total volume of the first solvent and the second solvent is 1 mg/mL-10 mg/mL; the molar ratio of the compound III to the compound V is 5:1 to 1:5, a step of; in the dynamic exchange photoreaction, the wavelength of light is 390nm, the power is 12W, the rotating speed is 600-800 rpm, the reaction temperature is room temperature, and the reaction time is 1-6 min.
In some embodiments, preferably, the molar ratio of compound iii to compound v is 5: 1. 4: 1. 3: 1. 2: 1. 1: 1. 1: 2. 1: 3. 1: 4. 1:5, further preferably 1:1, a step of; in the dynamic exchange photoreaction, the wavelength of light is 390nm, the power is 12W, the rotating speed is 700rpm, the reaction temperature is room temperature, and the reaction time is 1-6 min.
The first solvent in the first mixed solution is used for dissolving the reaction raw materials and has moderate viscosity; the second solvent is used in the second mixed solution in an amount to dissolve the reaction raw materials and has moderate viscosity.
The application of the amphiphilic diselenide compound in preparing the micelle for wrapping the medicine by self-assembly through an ultrasonic oscillation method as a nano medicine carrier is also within the protection scope of the invention.
Specifically, the specific method for preparing the micelle for wrapping the medicine by self-assembling the amphiphilic diselenide compound serving as the nano medicine carrier through an ultrasonic oscillation method comprises the following steps of:
(a) Dissolving a compound III in a first solvent to obtain a first mixed solution; dissolving the compound V in a second solvent to obtain a second mixed solution; uniformly mixing the first mixed solution and the second mixed solution, and then placing the mixed solution and the second mixed solution in a photoreactor for dynamic exchange photoreaction to obtain a reaction solution containing an amphipathic diselenide compound, namely a reaction solution containing a compound VI;
(b) Dissolving the medicine in a third solvent to obtain a third mixed solution; uniformly mixing the third mixed solution with the reaction solution containing the compound VI in the step (a) to obtain a fourth mixed solution; adding the fourth mixed solution into deionized water, carrying out ultrasonic vibration while adding the fourth mixed solution, continuing ultrasonic vibration after the fourth mixed solution is added, and standing after ultrasonic vibration is finished to obtain the micelle wrapping the medicine.
In some embodiments, in step (a), the first solvent is deuterated methanol or methanol; the second solvent is deuterated chloroform or dichloromethane; the mass-volume ratio of the total mass of the compound III and the compound V to the total volume of the first solvent and the second solvent is 1 mg/mL-10 mg/mL; the molar ratio of the compound III to the compound V is 5:1 to 1:5, a step of; in the dynamic exchange photoreaction, the wavelength of light is 390nm, the power is 12W, the rotating speed is 600-800 rpm, the reaction temperature is room temperature, and the reaction time is 1-6 min.
In some embodiments, preferably, in step (a), the molar ratio of compound iii to compound v is 5: 1. 4: 1. 3: 1. 2: 1. 1: 1. 1: 2. 1: 3. 1: 4. 1:5, further preferably 1:1, a step of; in the dynamic exchange photoreaction, the wavelength of light is 390nm, the power is 12W, the rotating speed is 700rpm, the reaction temperature is room temperature, and the reaction time is 1-6 min.
In some embodiments, in step (b), the third solvent is dichloromethane or ethanol; the medicine is a hydrophobic medicine; the hydrophobic drug is nile red; the mol volume ratio of the medicine in the third mixed solution to the third solvent is 1.89 mu mol-3.8 mu mol: 0.6-1.2 mL.
In some embodiments, the molar ratio of compound iii in step (a) to the drug in step (b) is 1:0.95 to 1.9; the mass to volume ratio of the total mass of the compound III, the compound V and the drug in the step (a) to the deionized water in the step (b) is 1mg:1mL.
In some embodiments, in step (b), the fourth mixed solution is added while performing ultrasonic vibration, and the ultrasonic power is 80W; and continuing ultrasonic oscillation after the fourth mixed solution is added, wherein the ultrasonic power is 80W, and the ultrasonic time is 10-20 min.
Further, the invention discloses galactose diselenide, and the structural formula of the galactose diselenide is shown as a formula III:
the invention further discloses a preparation method of the galactose diselenide, which comprises the following steps:
(1) The alpha-D-pentaacetyl galactose and 11-bromo-1-undecanol are subjected to etherification reaction under the action of a first catalyst, so that a compound I is prepared;
(2) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding the compound I obtained in the step (1) and a second catalyst into a reaction solution containing sodium diselenide, and performing a first coupling reaction to obtain a compound II;
(3) Performing hydrolysis reaction on the compound II obtained in the step (2) under the action of alkali to obtain galactose diselenide, namely a compound III;
wherein the above galactose diselenide, compound III, is denoted as (Gal-C11-Se) 2 。
In some embodiments, in step (1), the first catalyst is anhydrous zinc chloride; the molar ratio of the alpha-D-pentaacetyl galactose to the 11-bromo-1-undecanol is 1:1.33:1.33; the etherification reaction is carried out at the reaction temperature of 60-80 ℃ for 6-9 h.
In some embodiments, preferably, in step (1), the first catalyst is anhydrous zinc chloride; the molar ratio of the alpha-D-pentaacetyl galactose to the 11-bromo-1-undecanol is 1:1.33:1.33; the etherification reaction is carried out at the reaction temperature of 80 ℃ for 9 hours.
Wherein, the solvent used in the etherification reaction is anhydrous toluene; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
In some embodiments, in step (2), the second catalyst is tetrabutylammonium bromide; the molar ratio of the selenium powder to sodium borohydride to the compound I to the second catalyst is 1-1.5: 1 to 1.5:1:0.3; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 35-70 min; the first coupling reaction is carried out at room temperature for 8-12 h.
In some embodiments, preferably, in step (2), the second catalyst is tetrabutylammonium bromide; the molar ratio of the selenium powder to sodium borohydride to the compound I to the second catalyst is 1.5:1.5:1:0.3; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 65min; the first coupling reaction is carried out at room temperature for 12 hours.
Wherein the solvent used in the reduction reaction is ultrapure water; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the reduction reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
Wherein the solvent used in the first coupling reaction is tetrahydrofuran; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the first coupling reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
In some embodiments, in step (3), the base is sodium methoxide; the molar ratio of the compound II to the alkali is 1:1.8; the hydrolysis reaction is carried out at room temperature for 1-2 h.
In some embodiments, preferably, in step (3), the base is sodium methoxide; the molar ratio of the compound II to the alkali is 1:1.8; the hydrolysis reaction is carried out at room temperature for 1h.
Wherein, the solvent used in the hydrolysis reaction is a mixed solution of methanol and dichloromethane in any proportion, and the volume ratio of methanol to dichloromethane is preferably 1: 2; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Further, the invention discloses cholesterol diselenide, and the structural formula of the cholesterol diselenide is shown as formula V:
the invention further discloses a preparation method of the cholesterol diselenide, which comprises the following steps:
(i) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding 11-bromoundecanoic acid into a reaction solution containing sodium diselenide, and performing a second coupling reaction to prepare a compound IV;
(ii) The compound IV obtained in the step (i) and cholesterol are subjected to esterification reaction under the action of a dehydrating agent and a third catalyst to obtain cholesterol diselenide, namely a compound V;
wherein the cholesterol diselenide, compound V, is designated (Chol-C10-Se) 2 。
In some embodiments, in step (i), the molar ratio of selenium powder to sodium borohydride, 11-bromoundecanoic acid is 1:1:1, a step of; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 35-70 min; the second coupling reaction is carried out at room temperature for 8-12 h.
In some embodiments, preferably, in step (i), the molar ratio of selenium powder to sodium borohydride, 11-bromoundecanoic acid is 1:1:1, a step of; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 65min; and the second coupling reaction is carried out at room temperature for 12 hours.
Wherein the solvent used in the reduction reaction is ultrapure water; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the reduction reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
Wherein the solvent used in the second coupling reaction is tetrahydrofuran; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
Wherein the second coupling reaction is carried out under the protection of inert gas; the inert gas is preferably nitrogen.
In some embodiments, in step (ii), the dehydrating agent is N, N-dicyclohexylcarbodiimide; the third catalyst is 4-dimethylaminopyridine; the molar ratio of the compound IV to cholesterol, the dehydrating agent and the third catalyst is 1:2:2.1:0.03; the esterification reaction is carried out at room temperature for 10-15 h.
In some embodiments, preferably, in step (ii), the dehydrating agent is N, N-dicyclohexylcarbodiimide; the third catalyst is 4-dimethylaminopyridine; the molar ratio of the compound IV to cholesterol, the dehydrating agent and the third catalyst is 1:2:2.1:0.03; the esterification reaction is carried out at room temperature for 12 hours.
Wherein the solvent used in the esterification reaction is ultra-dry dichloromethane; the solvent is used in an amount to dissolve the solid raw materials in the reaction system and to have a moderate viscosity.
The use of the galactose diselenide or the cholesterol diselenide in the preparation of the amphiphilic diselenide compound is also within the scope of the invention.
The beneficial effects are that:
(1) The invention uses alpha-D-pentaacetyl galactose, 11-bromoundecanol, 11-bromoundecanoic acid, cholesterol and selenium powder as raw materials to respectively synthesize small molecule galactose diselenide ((Gal-C) containing diselenide bond and carbon chain 11 -Se) 2 ) And cholesterol diselenide ((Chol-C) 10 -Se) 2 ) Compared with the traditional method that expensive or toxic selenium nucleophilic reagent is needed for synthesizing diselenide, the method selects cheap and safe selenium powder, and greatly reduces potential safety hazard and synthesis cost.
(2) In the present invention, galactose diselenide ((Gal-C) 11 -Se) 2 ) The alpha-galactoside terminal configuration in the synthesis process can be turned into beta-galactoside terminal, so that the targeting tumor cell can be realizedForce; cholesterol diselenide ((Chol-C) 10 -Se) 2 ) In addition to acting as a hydrophobic end, providing a hydrophobic force for the self-assembly process, the biocompatibility of the carrier in vivo can also be increased.
(3) Galactose-cholesterol diselenide (Gal-C) in the present invention 11 -Se-Se-C 10 Chol) in the self-assembly process, galactose as hydrophilic end, cholesterol as hydrophobic end and intermediate carbon chain self-assemble in aqueous solution, spontaneously forming micelle with hydrophilic group towards water and lipophilic group inside; gal-C 11 -Se-Se-C 10 The Chol micelle can be uniformly dispersed in the aqueous solution, and the inner hydrophobic core can be used as a carrier for wrapping the anti-tumor hydrophobic drug in a nano drug-carrying system, so that the Chol micelle has good development and application prospects in tumor treatment.
(4) Gal-C in the present invention 11 -Se-Se-C 10 The Chol micelle, the hydrophilic galactose end is exposed on the surface of the micelle, so that specific tumor cells can be targeted, and the surface of the tumor cells is provided with receptors corresponding to the galactoside, so that the micelle can be better endocytosed into the tumor cells, and the anti-tumor effect is better achieved; the hydrophobic end cholesterol can enhance the biocompatibility and stability of the micelle and can avoid the damage of the micelle to normal cells.
(5) The preparation method provided by the invention has the advantages of simplicity in operation, environment friendliness, high economical efficiency and the like.
Drawings
The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings and detailed description.
FIG. 1 shows the compound I of example 1 1 H-NMR spectrum;
FIG. 2 is a diagram of compound I of example 1 13 C-NMR spectrum;
FIG. 3 shows the compound II of example 1 1 H-NMR spectrum;
FIG. 4 is a diagram of compound II of example 1 13 C-NMR spectrum;
FIG. 5 is a diagram of compound III of example 1 1 H-NMR spectrum;
FIG. 6 is a diagram of compound III of example 1 13 C-NMR spectrum;
FIG. 7 is a diagram of compound III of example 1 77 Se-NMR spectrum;
FIG. 8 is a HRMS mass spectrum of compound III of example 1;
FIG. 9 is a diagram of Compound V in example 2 1 H-NMR spectrum;
FIG. 10 is a diagram of compound V of example 2 13 C-NMR spectrum;
FIG. 11 is a diagram of Compound V in example 2 77 Se-NMR spectrum;
FIG. 12 is a HRMS mass spectrum of Compound V of example 2;
FIG. 13 shows two diselenides alone and two diselenides after 1min of illumination 77 Se-NMR spectrum;
FIG. 14 is a mass spectrum of MALDI-TOF-MS of two diselenide compounds alone and two diselenides after mixing and illumination for 6 min;
FIG. 15 shows Gal-C 11 -Se-Se-C 10 -Chol micelle size plot of dynamic light scattering DLS for different times;
FIG. 16 is Gal-C coated with hydrophobic nile red dye 11 -Se-Se-C 10 -fluorescence microscopy bright field and dark field images of Chol micelles;
FIG. 17 is Gal-C coated with hydrophobic nile red dye 11 -Se-Se-C 10 -a map of the responsive release of Chol micelles under different conditions;
FIG. 18 is Gal-C coated with hydrophobic nile red dye 11 -Se-Se-C 10 -two-photon laser confocal microscopy of Chol micelles targeting human ovarian cancer cell a2780 and human normal ovarian epithelial cell IOSE-80;
FIG. 19 is Gal-C without drug encapsulation 11 -Se-Se-C 10 Chol micelle electron microscopy.
Detailed Description
The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, are commercially available.
Phosphate Buffer (PBS) used in the examples of the present invention, ph=7.4, containing 135mM NaCl,4.7mM KCl,10mM Na 2 HPO 4 ，2mM NaH 2 PO 4 。
Example 1: synthesis of galactose diselenide (Compound III, (Gal-C) 11 -Se) 2 )
(1) Preparation of Compound I: adding a dry magneton into a 100mL single-neck flask, sequentially adding alpha-D-pentaacetyl galactose (2.5 g,6.40 mmoL), 11-bromo-1-undecanol (2.1352 g,8.50 mmoL) and 4mL anhydrous toluene, placing the reaction flask into an oil bath at 80 ℃ for stirring, adding anhydrous zinc chloride (1.160 g,8.5 mmoL) dried in advance for 1h after the solid in the flask is completely dissolved, stirring in the oil bath at 80 ℃ for reacting until the reaction liquid turns from colorless to yellow, and stirring for etherification for 9h; after the reaction, the clear yellow reaction solution was transferred to a magnetic stirrer at room temperature, ethyl acetate (20 mL) and saturated sodium bicarbonate aqueous solution (20 mL) were added in this order to the reaction solution, and the mixture was quenched and stirred for 10min; then removing powder by vacuum suction filtration; extracting the reaction solution with ultrapure water for 3 times, and taking an organic phase; drying with anhydrous magnesium sulfate, and removing powder by vacuum filtration again; finally, the solvent was removed by a vacuum rotary evaporator, and the oily yellow crude product obtained was purified by column chromatography (eluent petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate was 20:1-10:1; 10:1 output point) to give compound i, 1.2436g, in 33.49% yield.
For compounds I 1 H-NMR 13 C-NMR characterization, specifically nuclear magnetic hydrogen spectrogram is shown in figure 1, and nuclear magnetic carbon spectrogram is shown in figure 2.
(2) Preparation of Compound II: adding a dry magneton into a 100mL three-necked flask in a magnetic stirrer at room temperature, vacuumizing, introducing nitrogen, and repeating for three times; all conditions at the time of loading were under nitrogen atmosphere. Selenium powder (0.1269 g,1.6076 mmo) was added to the flask in sequenceL) and ultrapure water (6 mL); then adding ice bath condition (ice cubes), adding sodium borohydride (0.1216 g,3.2151 mmol) into the reaction system at 0 ℃, stirring and reacting until the reaction liquid becomes clear from black, wherein the reaction time is 20min; then adding selenium powder (0.1269 g,1.6076 mmoL) into the reaction system, moving the three-neck flask to an oil bath kettle at 50 ℃, stirring for reduction reaction until the reaction liquid turns from black to red, and reacting for 45min to obtain a reaction liquid containing sodium diselenide; then, to the reaction solution containing sodium diselenide, a tetrahydrofuran solution (5 mL) containing Compound I (1.2436 g,2.1434 mmoL) and a tetrahydrofuran solution (2 mL) containing tetrabutylammonium bromide (0.2073 g,0.6430 mmoL) were sequentially added, and the reaction solution was stirred at room temperature until the reaction solution became dark green from reddish wine, and a first coupling reaction was carried out for 12 hours; after the reaction is finished, firstly removing unreacted selenium powder by vacuum filtration, then screwing off tetrahydrofuran, and then extracting for three times by using dichloromethane and saturated saline water, and taking a yellow organic phase; then the organic phase is dried by anhydrous magnesium sulfate, and the powder is removed by vacuum filtration again; finally, the solvent was removed by vacuum rotary evaporator, and the oily yellow crude product was purified by thin layer chromatography (petrol ether and ethyl acetate as the developing solvent, and petrol ether and ethyl acetate in a 3:2 volume ratio) to give compound II, 0.9981g, in 80.26% yield. For compounds II 1 H-NMR 13 C-NMR characterization, specifically nuclear magnetic hydrogen spectrum shown in figure 3, and nuclear magnetic carbon spectrum shown in figure 4.
(3) A25 mL single-necked flask was charged with dry magneton in a magnetic stirrer at room temperature, and a mixed solution of methanol (3 mL) and methylene chloride (6 mL) containing Compound II (0.9981 g,0.858 mmoL) was added to the flask; then, sodium methoxide methanol solution (86. Mu.L, 1.5457 mmoL) was added thereto, and the mixture was stirred at room temperature to carry out hydrolysis reaction for 1 hour; after completion of the reaction, the reaction mixture was quenched with acetic acid (47.6. Mu.L) for 10min; and (5) screwing off the reaction liquid to obtain galactose diselenide, namely the compound III. For compounds III 1 H-NMR、 13 C-NMR、 77 Se-NMR and HRMS characterization, specific nuclear magnetic hydrogen spectrogram is shown in fig. 5, nuclear magnetic carbon spectrogram is shown in fig. 6, nuclear magnetic selenium spectrogram is shown in fig. 7, mass spectrogram is shown in fig. 8, and mass spectrum data are shown in the following: HRMS [ M+Na ]]calculated for C 34 H 66 NaO 12 Se 2 + 849.2777u,found 849.2783u.
Example 2: synthesis of cholesterol diselenide (Compound V, (Chol-C) 10 -Se) 2 )
(i) In a magnetic stirrer at room temperature, a 100mL three-necked flask was charged with dry magneton, evacuated, and purged with nitrogen, and repeated three times. All conditions at the time of loading were under nitrogen atmosphere. Selenium powder (0.75 g,9.5 mmol) and ultrapure water (20 mL) are added in sequence; then adding ice bath condition (ice cubes), adding sodium borohydride (0.75 g,20 mmol) into the reaction system at 0 ℃, and stirring to react until the reaction liquid becomes clear from black, wherein the reaction time is 20min; then adding selenium powder (0.75 g,9.5 mmol) into the reaction system, transferring the three-necked flask to an oil bath pot at 50 ℃, stirring and carrying out reduction reaction until the reaction liquid changes from black to red, and the reaction time is 45min, thus obtaining a reaction liquid containing sodium diselenide;
Then adding tetrahydrofuran solution (10 mL) containing 11-bromoundecanoic acid (5.03 g,19 mmol) into the reaction solution containing sodium diselenide in turn, stirring at room temperature, and performing a second coupling reaction until the reaction solution turns from wine red to yellow, wherein the reaction time is 12h; after the reaction is finished, firstly reducing the pressure and filtering to remove unreacted selenium powder; extracting with dichloromethane for 1-2 times; extracting with 0.1M hydrochloric acid for 2 times, and collecting yellow organic phase; then the organic phase is dried by anhydrous magnesium sulfate, and the powder is removed by vacuum filtration again; finally, the solvent is removed by a vacuum rotary evaporator, and the obtained oily yellow crude product is subjected to a recrystallization method to obtain the crude product. The specific method is that the poor solvent is recrystallized; the mixture was dissolved with a small amount of methylene chloride, and then petroleum ether, a poor solvent having a volume 50 times larger than that of the good solvent, was added thereto, and left to stand overnight to give a crude product IV, 1.6838g.
(ii) In a 250mL single-neck flask was added dry magneton, and under ice bath, compound IV (0.5285 g,1 mmol) and ultra-dry dichloromethane (30 mL) were added sequentially; next, N-dicyclohexylcarbodiimide (0.433)3g,2.1 mmol) of dichloromethane (5 mL), and stirring for 10min; then, methylene chloride (2 mL) containing 4-dimethylaminopyridine (0.0037 g,0.03 mmoL) was added dropwise, and the reaction was stirred for 15min; finally, cholesterol (0.7733 g,2 mmol) was added and the mixture was stirred at room temperature for esterification for 12h; after the reaction is finished, firstly removing unreacted substances by vacuum suction filtration; extracting with saturated saline for three times, and collecting yellow organic phase; drying with anhydrous magnesium sulfate, and removing powder by vacuum filtration again; finally, the solvent was removed by a vacuum rotary evaporator, and the resulting crude yellow solid was purified by column chromatography (eluent n-hexane and dichloromethane with a volume ratio of n-hexane to dichloromethane of 10:1 to 4:1; 4:1 output point) to give cholesterol diselenide ether, compound v, 0.9062g, yield 71.53%. For compounds V 1 H-NMR、 13 C-NMR、 77 Se-NMR and HRMS, a specific nuclear magnetic hydrogen spectrum is shown in fig. 9, a nuclear magnetic carbon spectrum is shown in fig. 10, a nuclear magnetic selenium spectrum is shown in fig. 11, and a mass spectrum is shown in fig. 12, and specific mass spectrum data are as follows: HRMS [ M+Na + ]calculated for C 76 H 130 NaO 4 Se 2 + 1289.8192u,found 1289.8163u.
Example 3: galactose diselenide ((Gal-C11-Se) 2 ) And cholesterol diselenide ((Chol-C10-Se) 2 ) Preparation of galactose-cholesterol diselenether (Gal-C11-Se-Se-C10-Chol) by dynamic exchange of diselenide bonds in a photoreactor
(1) Firstly, galactose diselenide (0.0160 g,0.02mmol, prepared in example 1) is weighed and dissolved in deuterated methanol (1 mL) to obtain a first mixed solution; cholesterol diselenide (0.0240 g,0.02mmol, prepared in example 2) was weighed and dissolved in deuterated chloroform (3 mL) to give a second mixture; and (3) uniformly mixing the first mixed solution and the second mixed solution, adding the mixed solution into a photoreaction tube, and carrying out dynamic exchange reaction for 1min at room temperature under the illumination condition of 390nm in wavelength, 12W in power and 700rpm in rotating speed.
Taking deuterated methanol solution containing galactose diselenide (0.0300 g,0.36 mmol), deuterated chloroform solution containing cholesterol diselenide ether (0.0300 g,0.024 mmol), and reaction solution obtained by mixing two diselenide ethers and illuminating for 1min 77 Se-NMRThe specific verification pattern is shown in fig. 13, wherein fig. 13a is a deuterated methanol solution detection chart containing galactose diselenide, fig. 13b is a deuterated chloroform solution detection chart containing cholesterol diselenide, and fig. 13c is a detection chart of illumination for 1min after two diselenides are uniformly mixed; as can be seen from FIG. 13, comparing the peak position changes of the selenium spectrum before and after mixing galactose diselenide and cholesterol diselenide, it can be demonstrated that dynamic exchange of diselenide bonds can occur between two diselenides after 1min of illumination in the photoreactor.
(2) Firstly, galactose diselenide (0.00160 g,0.002mmol, prepared in example 1) is weighed and dissolved in methanol (1 mL) to obtain a first mixed solution; cholesterol diselenide (0.0024 g,0.002mmol, prepared in example 2) was weighed and dissolved in dichloromethane (3 mL) to give a second mixture; and (3) uniformly mixing the first mixed solution and the second mixed solution, adding the mixed solution into a photoreaction tube, and carrying out dynamic exchange reaction for 6min at room temperature under the illumination condition of 390nm in wavelength, 12W in power and 700rpm in rotating speed.
Separately taking methanol solution containing galactose diselenide (0.0015 g), dichloromethane solution containing cholesterol diselenide (0.0015 g) and mixed solution of two diselenides after illumination for 6min, and performing MALDI-TOF-MS verification, wherein specific verification data is shown in figure 14; as can be seen from fig. 14, after the two diselenides are illuminated by the light of the photoreactor for 6min, the molecular weight of the amphiphilic diselenide compound corresponding to the diselenide compound after the exchange can be found, and specific mass spectrum data are as follows: MALDI-TOF-MS [ M+Na ] + ]calculated for C 55 H 98 NaO 8 Se 2 + 1069.5484u,found 1069.7964u。
Example 4: galactose-cholesterol diselenide (Gal-C) encapsulated with hydrophobic drug 11 -Se-Se-C 10 -Chol) micelle preparation and pair H of micelles 2 O 2 Responsiveness to Glutathione (GSH) and beta-galactosidase
(1) Preparation of galactose-cholesterol diselenide (Gal-C11-Se-Se-C10-Chol) micelle
(a) Firstly, the mol ratio of galactose diselenide to cholesterol diselenide is 1:1, 0.0016g of galactose diselenide (compound III,0.002mmol, prepared in example 1) was weighed and dissolved in methanol (1 mL) to obtain a first mixed solution; 0.0024g cholesterol diselenide (compound V,0.002mmol, prepared in example 2) was weighed and dissolved in dichloromethane (3 mL) to give a second mixture; and (3) placing the first mixed solution and the second mixed solution in a photoreactor, wherein the wavelength is 390nm, the power is 12W, the rotating speed is 700rpm, and the dynamic exchange reaction is carried out for 6min at room temperature, so that the dynamic exchange of diselenide bonds is completed, and the reaction solution containing the amphipathic diselenide compound, namely the reaction solution containing the compound VI, is obtained.
(b) Then, 0.0012g of nile red dye (3.8 mu moL) was weighed and dissolved in 1.2mL of methylene chloride to obtain a third mixed solution; uniformly mixing the third mixed solution with the reaction solution containing the compound VI to obtain a fourth mixed solution; dropwise adding the fourth mixed solution into 5.2mL of deionized water, and simultaneously carrying out ultrasonic vibration (the ultrasonic power is 80W) while adding the fourth mixed solution, and continuing ultrasonic vibration for 10min (the ultrasonic power is 80W) after the addition of the fourth mixed solution is finished; and standing for one night after the ultrasonic treatment is finished, thus obtaining the micelle solution for coating the hydrophobic nile red dye.
(2) Micelle pair H of encapsulated nile red dye 2 O 2 Responsiveness to Glutathione (GSH) and beta-galactosidase
First, 194. Mu.L of a micelle solution (prepared in step (1) of example 4) coated with a nile red dye was added to a 96-well plate, and four groups of 3 groups of the micelle solution were used in parallel experiments, and the four groups were a blank group of PBS and 25mM H, respectively 2 O 2 25mM GSH and 1 μg/mL β -gal; then 6. Mu.L of PBS and H were added to each group 2 O 2 GSH and β -gal, with a final volume of 200 μl per well; then placing the 96-well plate into a constant-temperature shaking box at 37 ℃ for incubation for 30min; and finally, measuring the fluorescence intensity released by the nile red micelle in response to different conditions. From FIG. 17, it can be seen that Gal-C, which encapsulates hydrophobic nile red dye 11 -Se-Se-C 10 Chol micelles have a higher response to 25mM glutathione and 1. Mu.g/mL. Beta. -gal.
Example 5: galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 -Chol) micelles detecting hydrodynamic diameters of different time periods by Dynamic Light Scattering (DLS)
(1) Galactose-cholesterol diselenideEthers (Gal-C) 11 -Se-Se-C 10 Preparation of Chol) micelles
Firstly, the mol ratio of galactose diselenide to cholesterol diselenide is 1:1, 0.0016g of galactose diselenide (compound III,0.002mmol, prepared in example 1) was weighed and dissolved in methanol (1 mL) to obtain a first mixed solution; 0.0024g cholesterol diselenide (compound V,0.002mmol, prepared in example 2) was weighed and dissolved in dichloromethane (3 mL) to give a second mixture; placing the first mixed solution and the second mixed solution in a photoreactor, and dynamically exchanging and reacting for 6 minutes at room temperature at the wavelength of 390nm and the power of 12W and the rotating speed of 700rpm to finish dynamic exchange of diselenide bonds, thereby obtaining a reaction solution containing amphipathic diselenide compounds, namely a reaction solution containing compounds VI; dropwise adding the reaction solution containing the compound VI into 4mL of deionized water, and simultaneously carrying out ultrasonic vibration (the ultrasonic power is 80W) while adding the reaction solution containing the compound VI, and continuing ultrasonic vibration for 10min (the ultrasonic power is 80W) after the addition of the reaction solution containing the compound VI is finished; standing overnight after ultrasonic treatment to obtain galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 Chol) micelle solution.
(2) Galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 -Chol) determination of micelle hydrodynamic diameter
Taking 3mL of galactose-cholesterol diselenide (Gal-C11-Se-Se-C10-Chol) micelle solution (prepared in the step (1) of the example 5), placing the sample pool in a Markov particle size sample pool, setting the temperature to 25 ℃, and measuring the particle sizes of different time periods respectively at 0h, 1h, 2h, 3h, 4h, 5h, 6h and 12h, wherein each time period is measured three times; as can be seen from FIG. 15, gal-C 11 -Se-Se-C 10 Chol micelles are stable well within 12 h.
(3) Galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 Transmission Electron Microscope (TEM) morphology characterization of Chol) micelles
10. Mu.L galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 -Chol) the upper aqueous layer of the micellar solution (prepared in step (1) of example 5) was dropped onto a common copper mesh and repeated3 times; and finally, naturally airing and then loading the sample into a machine for testing. As can be seen from FIG. 19, galactose-cholesterol diselenide (Gal-C 11 -Se-Se-C 10 Chol) micelle is solid spherical micelle, has a particle size of about 200nm, and is uniformly distributed.
Example 6: galactose-cholesterol diselenide (Gal-C) 11 -Se-Se-C 10 -Chol) micelle-encapsulated hydrophobic nile red dye fluorescence microscopy bright field and dark field images
10. Mu.L of galactose-cholesterol diselenide ether (Gal-C) coated with hydrophobic nile red dye was taken 11 -Se-Se-C 10 Chol) micelle solution (prepared in step (1) of example 4) was placed on a special coverslip, dried naturally and then measured on a machine. From FIG. 16, it can be seen that Gal-C can be demonstrated by comparing pictures taken in the bright field with those taken in the dark field, by fluorescence of micelles in which hydrophobic nile red dye is encapsulated under excitation at 530nm (nile red: ex=528 nm, em=576 nm) 11 -Se-Se-C 10 The core of the Chol micelle may encapsulate the hydrophobic drug.
Example 7: galactose-cholesterol diselenide ether (Gal-C) encapsulating nile red dye 11 -Se-Se-C 10 Two-photon laser confocal microscopy image of Chol) micelle targeted human ovarian cancer cells A2780 and human normal ovarian epithelial cells IOSE-80
Galactose-cholesterol diselenide ether (Gal-C) encapsulating nile red dye 11 -Se-Se-C 10 Chol) micelle solution (prepared in step (1) of example 4) was incubated with human ovarian cancer cells a2780 and human normal ovarian epithelial cells IOSE-80, respectively, for 6h, and the targeting ability of the micelle to different cells was photographed by a two-photon laser confocal microscope. As can be seen from FIG. 18, fluorescence aggregation was found around human ovarian cancer cells A2780, which verifies that beta-galactoside on micelle can be specifically bound with galectin on the surface of A2780 cells, while no fluorescence appears around human normal ovarian epithelial cells IOSE-80, indicating Gal-C coating hydrophobic nile red dye 11 -Se-Se-C 10 Chol micelles are targeted.
The invention provides a diselenide with targeting and responsiveness, a preparation method and an application thought and method thereof, and a method for realizing the technical scheme is a plurality of methods and approaches, the above is only a preferred embodiment of the invention, and it should be pointed out that a plurality of improvements and modifications can be made by one of ordinary skill in the art without departing from the principle of the invention, and the improvements and modifications are also considered as the protection scope of the invention. The components not explicitly described in this embodiment can be implemented by using the prior art.

Claims (21)

1. The amphipathic diselenide compound is characterized by having a structural formula shown in a formula VI:
2. the method for preparing an amphiphilic diselenide compound as claimed in claim 1, comprising the steps of: dissolving a compound III in a first solvent to obtain a first mixed solution; dissolving the compound V in a second solvent to obtain a second mixed solution; uniformly mixing the first mixed solution and the second mixed solution, then placing the mixed solution into a photoreactor, performing dynamic exchange photoreaction to obtain a reaction solution containing an amphipathic diselenide compound, and performing aftertreatment on the reaction solution to obtain the amphipathic diselenide compound, namely a compound VI;
3. The preparation method according to claim 2, wherein the preparation method of the compound iii comprises the steps of:
(1) The alpha-D-pentaacetyl galactose and 11-bromo-1-undecanol are subjected to etherification reaction under the action of a first catalyst, so that a compound I is prepared;
(2) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding the compound I obtained in the step (1) and a second catalyst into a reaction solution containing sodium diselenide, and performing a first coupling reaction to obtain a compound II;
(3) Performing hydrolysis reaction on the compound II obtained in the step (2) under the action of alkali to obtain galactose diselenide, namely a compound III;
4. the preparation method according to claim 2, wherein the preparation method of the compound v comprises the steps of:
(i) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding 11-bromoundecanoic acid into a reaction solution containing sodium diselenide, and performing a second coupling reaction to prepare a compound IV;
(ii) The compound IV obtained in the step (i) and cholesterol are subjected to esterification reaction under the action of a dehydrating agent and a third catalyst to obtain cholesterol diselenide, namely a compound V;
5. The method of claim 2, wherein the first solvent is deuterated methanol or methanol; the second solvent is deuterated chloroform or dichloromethane; the mass-volume ratio of the total mass of the compound III and the compound V to the total volume of the first solvent and the second solvent is 1 mg/mL-10 mg/mL; the molar ratio of the compound III to the compound V is 5:1 to 1:5, a step of; in the dynamic exchange photoreaction, the wavelength of light is 390nm, the power is 12W, the rotating speed is 600-800 rpm, the reaction temperature is room temperature, and the reaction time is 1-6 min.
6. The use of the amphiphilic diselenide compound of claim 1 as a nano drug carrier for preparing drug-coated micelles by self-assembly through an ultrasonic oscillation method.
7. The application of claim 6, wherein the specific method for preparing the micelle for wrapping the drug by self-assembling the amphiphilic diselenide compound serving as the nano drug carrier through an ultrasonic oscillation method comprises the following steps:
(a) Dissolving a compound III in a first solvent to obtain a first mixed solution; dissolving the compound V in a second solvent to obtain a second mixed solution; uniformly mixing the first mixed solution and the second mixed solution, and then placing the mixed solution and the second mixed solution in a photoreactor for dynamic exchange photoreaction to obtain a reaction solution containing an amphipathic diselenide compound, namely a reaction solution containing a compound VI;
(b) Dissolving the medicine in a third solvent to obtain a third mixed solution; uniformly mixing the third mixed solution with the reaction solution containing the compound VI in the step (a) to obtain a fourth mixed solution; adding the fourth mixed solution into deionized water, carrying out ultrasonic vibration while adding the fourth mixed solution, continuing ultrasonic vibration after the fourth mixed solution is added, and standing after ultrasonic vibration is finished to obtain the micelle wrapping the medicine.
8. The use according to claim 7, wherein in step (a), the first solvent is deuterated methanol or methanol; the second solvent is deuterated chloroform or dichloromethane; the mass-volume ratio of the total mass of the compound III and the compound V to the total volume of the first solvent and the second solvent is 1 mg/mL-10 mg/mL; the molar ratio of the compound III to the compound V is 5:1 to 1:5, a step of; in the dynamic exchange photoreaction, the wavelength of light is 390nm, the power is 12W, the rotating speed is 600-800 rpm, the reaction temperature is room temperature, and the reaction time is 1-6 min.
9. The use according to claim 7, wherein in step (b), the third solvent is dichloromethane or ethanol; the medicine is a hydrophobic medicine; the hydrophobic drug is nile red; the mol volume ratio of the medicine in the third mixed solution to the third solvent is 1.89 mu mol-3.8 mu mol: 0.6-1.2 mL.
10. The use according to claim 7, wherein the molar ratio of compound iii in step (a) to the medicament in step (b) is 1:0.95 to 1.9; the mass to volume ratio of the total mass of the compound III, the compound V and the drug in the step (a) to the deionized water in the step (b) is 1mg:1mL.
11. The use according to claim 7, wherein in step (b), the fourth mixed solution is added and simultaneously subjected to ultrasonic vibration, the ultrasonic power being 80W; and continuing ultrasonic oscillation after the fourth mixed solution is added, wherein the ultrasonic power is 80W, and the ultrasonic time is 10-20 min.
12. The galactose diselenide is characterized in that the structural formula of the galactose diselenide is shown as a formula III:
13. the method for preparing galactose diselenide of claim 12, comprising the steps of:
(1) The alpha-D-pentaacetyl galactose and 11-bromo-1-undecanol are subjected to etherification reaction under the action of a first catalyst, so that a compound I is prepared;
(2) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding the compound I obtained in the step (1) and a second catalyst into a reaction solution containing sodium diselenide, and performing a first coupling reaction to obtain a compound II;
(3) Performing hydrolysis reaction on the compound II obtained in the step (2) under the action of alkali to obtain galactose diselenide, namely a compound III;
14. the method of claim 3 or claim 13, wherein in step (1), the first catalyst is anhydrous zinc chloride; the molar ratio of the alpha-D-pentaacetyl galactose to the 11-bromo-1-undecanol is 1:1.33:1.33; the etherification reaction is carried out at the reaction temperature of 60-80 ℃ for 6-9 h.
15. The process according to claim 3 or claim 13, wherein in step (2), the second catalyst is tetrabutylammonium bromide; the molar ratio of the selenium powder to sodium borohydride to the compound I to the second catalyst is 1-1.5: 1 to 1.5:1:0.3; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 35-70 min; the first coupling reaction is carried out at room temperature for 8-12 h.
16. The process according to claim 3 or claim 13, wherein in step (3), the base is sodium methoxide; the molar ratio of the compound II to the alkali is 1:1.8; the hydrolysis reaction is carried out at room temperature for 1-2 h.
17. The cholesterol diselenide is characterized in that the structural formula of the cholesterol diselenide is shown as a formula V:
18. the method for preparing cholesterol diselenide of claim 17, comprising the steps of:
(i) Carrying out reduction reaction on the selenium powder, sodium borohydride and water to obtain a reaction solution containing sodium diselenide; adding 11-bromoundecanoic acid into a reaction solution containing sodium diselenide, and performing a second coupling reaction to prepare a compound IV;
(ii) The compound IV obtained in the step (i) and cholesterol are subjected to esterification reaction under the action of a dehydrating agent and a third catalyst to obtain cholesterol diselenide, namely a compound V;
19. the method of claim 4 or claim 18, wherein in step (i), the molar ratio of selenium powder to sodium borohydride, 11-bromoundecanoic acid is 1:1:1, a step of; the reduction reaction is carried out at the reaction temperature of 0-50 ℃ for 35-70 min; the second coupling reaction is carried out at room temperature for 8-12 h.
20. The process according to claim 4 or claim 18, wherein in step (ii), the dehydrating agent is N, N-dicyclohexylcarbodiimide; the third catalyst is 4-dimethylaminopyridine; the molar ratio of the compound IV to cholesterol, the dehydrating agent and the third catalyst is 1:2:2.1:0.03; the esterification reaction is carried out at room temperature for 10-15 h.
21. Use of galactose diselenide according to claim 12 or cholesterol diselenide according to claim 17 for the preparation of an amphiphilic diselenide compound according to claim 1.
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