阅读说明：本技术 一种聚乙二醇衍生物及其制备方法和用途 (Polyethylene glycol derivative and preparation method and application thereof ) 是由 王晓 于 2020-06-15 设计创作，主要内容包括：本发明提供了一种聚乙二醇衍生物及其制备方法和用途,所述聚乙二醇衍生物包括含有脂肪链的基团和聚乙二醇,所述含有脂肪链的基团与聚乙二醇通过连接基团连接,其中所述含有脂肪链的基团中的至少一个基团是磷脂类基团。其中疏水性部分(即亲油基团)可以通过改变脂肪链链长、饱和度以及脂肪链数量来调节疏水部分的体积和缔合作用强度,而亲水部分的体积可以通过选择聚乙二醇高分子链长和分支数量来调节,两者协同配合,可以得到不同胶束尺寸的、稳定的胶束。(The invention provides a polyethylene glycol derivative, a preparation method and application thereof, wherein the polyethylene glycol derivative comprises a group containing a fatty chain and polyethylene glycol, the group containing the fatty chain is connected with the polyethylene glycol through a connecting group, and at least one group in the group containing the fatty chain is a phospholipid group. The volume of the hydrophobic part and the association strength can be adjusted by changing the chain length, the saturation and the number of the fatty chains of the hydrophobic part, the volume of the hydrophilic part can be adjusted by selecting the chain length and the number of branches of the polyethylene glycol polymer, and the two are cooperated to obtain stable micelles with different micelle sizes.)

1. The polyethylene glycol derivative has a structure of PEG- (L)₁-R₁)_nWherein PEG is polyethylene glycol group, L₁Is a divalent linking group, n is an integer between 2 and 10, R₁Same or different, independently selected from the group consisting of phospholipid groups, C_8-40A hydrocarbyl group, and wherein at least one R is₁Is a phospholipid group;

wherein the phospholipid group has a structure shown in a formula (I) or a formula (II),

or

Wherein R is₂、R₃The same or different, are independently selected from C_8-40Hydrocarbyl represents a bond.

2. The polyethylene glycol derivative according to claim 1, wherein the linking group L₁The same or different, said linking group L₁Contains the following groups: -CO-NH-, -CO-O-, -CS-O-, -NH-CO-NH-, -CO-NH-CO-, -O-, -S-S-, -NH-,an azido-alkynyl cycloaddition linking group, a tetrazinyl-alkenyl cycloaddition linking group.

3. The polyethylene glycol derivative according to claim 1, wherein the linking group L₁Comprises the following steps: -CO-CH₂-CH₂-CO-NH-CH₂-CH₂-、-CO-CH₂-CH₂-CO-NH-、-CH₂-CO-NH-CH₂-CH₂-、-CH₂-CO-NH-、-S-S-、。

4. The polyethylene glycol derivative according to claim 1, wherein PEG is a linear polyethylene glycol group or a multi-arm polyethylene glycol group which is a three-arm polyethylene glycol group, a four-arm polyethylene glycol group, a five-arm polyethylene glycol group, a six-arm polyethylene glycol group, a seven-arm polyethylene glycol group, an eight-arm polyethylene glycol group, a nine-arm polyethylene glycol group, or a ten-arm polyethylene glycol group; the multi-arm polyethylene glycol group comprises a polyethylene glycol repeating unit and a branched core structure, wherein the branched core structure is selected from glycerol, glycidyl ether, ethylene glycol glycidyl ether, pentaerythritol, diglycidyl pentaerythritol ether, triglycidyl ether, gallic acid ether or ethylene glycol gallic acid ether.

5. A polyethylene glycol derivative is prepared by reacting the following raw materials:

(a) one or more kinds of phospholipid compounds, and (b) polyethylene glycol PEG- (Y) with Y as terminal group_nAnd optionally one or more than two (c) compounds R containing an aliphatic chain₄-X₁Wherein the phospholipid compound is glycerophospholipid or lysophospholipid, and the amino group or hydroxyl group in the phospholipid compound is reacted with (b) PEG- (Y)_nY in (b) is reacted with (c) R₄-X₁X in (1)₁And (b) PEG- (Y)_nReacting Y in the step (1);

wherein, PEG is a linear polyethylene glycol group or a multi-arm polyethylene glycol group, the multi-arm polyethylene glycol group is a three-arm polyethylene glycol group, a four-arm polyethylene glycol group, a five-arm polyethylene glycol group, a six-arm polyethylene glycol group, a seven-arm polyethylene glycol group, an eight-arm polyethylene glycol group, a nine-arm polyethylene glycol group or a ten-arm polyethylene glycol group, and n is an integer between 2 and 10;

the radicals Y being identicalOr different, independently selected from-R₅-a, said a being a reactive group: -COOH, amino, -OH, -SH, -C.ident.N, -C.ident.C, -C = C,N-Hydroxyl succinimide ester group, maleimide group, dithiopyridyl group, vinyl sulfone group, isocyanate group and tetrazine group; the R is₅Is absent or a divalent linking group;

the group X₁The same or different, are independently selected from the following reactive groups: -COOH, amino, -OH, -SH, -C.ident.N, -C.ident.C, -C = C,N-Any one of a hydroxysuccinimide ester group, a maleimide group, a dithiopyridyl group, a vinylsulfone group, an isocyanate group and a tetrazine group;

R₄is C_8-40A hydrocarbyl group.

6. The polyethylene glycol derivative according to claim 5, wherein said glycerophospholipid is Phosphatidylethanolamine (PE), Phosphatidylserine (PS), Phosphatidylglycerol (PG) or Phosphatidic Acid (PA), and said lysophospholipid is lysophosphatidylcholine (L PC), lysophosphatidylethanolamine (L PE), lysophosphatidylinositol (L PI) or lysophosphatidic acid (L PA).

7. The polyethylene glycol derivative according to claim 6, wherein the phosphatidylethanolamine is selected from the group consisting of dihexanoylphosphatidylethanolamine, dioctanoylphosphatidylethanolamine, didecanoylphosphatidylethanolamine, dilauroylphosphatidylethanolamine (D L PE), Dimyristoylphosphatidylethanolamine (DMPE), Dipalmitoylphosphatidylethanolamine (DPPE), Distearoylphosphatidylethanolamine (DSPE), Dioleoylphosphatidylethanolamine (DOPE), Dicambaoylphosphatidylethanolamine (DEPE), 1, 2-Docosahexaenoyl (DHA) phosphatidylethanolamine, 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), 1-stearoyl-2-oleoylphosphatidylethanolamine (SOPE);

the lysophosphatidylethanolamine is selected from myristoyl lysophosphatidylethanolamine (L MPE), palmitoyl lysophosphatidylethanolamine (L PPE), and stearoyl lysophosphatidylethanolamine (L SPE).

8. A process for the preparation of a polyethylene glycol derivative according to any one of claims 1 to 7, said process comprising the steps of:

the phospholipid compound (a) and the polyethylene glycol PEG- (Y) with Y as the end group (b)_nAnd optionally (c) a compound R containing an aliphatic chain₄-X₁Mixing and reacting to obtain said polyethylene glycol derivative, wherein PEG is as defined in any one of claims 2 to 7, Y, R₄、X₁The method according to any one of claims 5 to 7.

9. A micelle comprising the polyethylene glycol derivative according to any one of claims 1 to 7.

10. Use of the micelle of claim 9 in the coating and/or delivery of hydrophobic compounds.

Technical Field

The invention relates to a polyethylene glycol derivative, a preparation method thereof and application thereof in coating and delivering a hydrophobic compound, belonging to the technical field of lipoid molecules.

Background

After the surfactant containing hydrophilic and lipophilic groups reaches and exceeds a certain concentration in solution, the molecules self-associate into colloid-sized aggregate particles called micelles, which is called Critical Micelle Concentration (CMC). Micelle formation is an entropy-driven process. When a single surfactant molecule is completely surrounded by water molecules in water, the degree of freedom of the water molecules at the contact interface of the oleophilic group and the water molecules is rapidly reduced, so that the entropy of a mixed system is reduced, and the mixed system becomes a thermodynamically unstable state. To maintain the overall entropy of the system, the surfactant molecules first occupy the solution surface with their lipophilic groups facing the air, exhibiting surface adsorption. If the concentration of the surfactant continues to increase, the surfactant molecules in the solution avoid entropy reduction by aggregating with each other after the surface adsorption reaches saturation. At the moment, the hydrophilic group faces to water molecules, and the lipophilic groups are mutually associated to form molecular aggregates, so that the low-entropy state of the water molecules at the contact interface of the lipophilic groups and the water is reduced, and the thermodynamic stability of the system is maintained. Thus, when the surfactant exceeds the critical colloidal concentration, micelles spontaneously form in solution and are in dynamic equilibrium with free surfactant molecules.

The structural characteristics of the micelle are that the monolayer aggregate of the amphiphilic surfactant molecules is mostly spherical, and can also form a column under a specific molecular structure. Since micelles do not contain a bilayer structure and do not have an inner vesicle cavity, they have a smaller structural size than liposomes. The hydration radius of the micelles formed by the single fatty chain surfactant is generally 2nm to 20nm, which is less than the lower limit of the liposome size and is more permeable to skin and tissues. The hydrophobic core of the micelle makes it possible to encapsulate poorly soluble hydrophobic compounds in the aqueous phase. The micelle is in dynamic balance in the solution, so that the micelle can quickly release insoluble active ingredients and can be used for a delivery system of medicines and cosmetics. Compared with a liposome delivery system, the structural characteristics of the molecular arrangement of the amphiphilic molecules in a monolayer make the stability of the micelle inferior to that of the liposome. When the micelle contacts with the surface skin tissue, the micelle can be rapidly disintegrated, so that the effective components wrapped by the micelle system can not be released for a long time. Free surfactants in micellar systems can disrupt cell membranes, causing allergic reactions in biological tissues and skin.

Amphiphilic molecules with the volume of the hydrophilic group close to that of the lipophilic group are easy to form liposome, and the surfactant with the volume of the hydrophilic group larger than that of the lipophilic group is easy to form micelle. Common single chain fatty acid surfactants promote micelle formation by reducing the volume of the lipophilic group. The micelle formed by the surfactant consisting of a single fatty chain has weak association of oleophilic groups, and surfactant molecules in the micelle are easy to leave the micelle through dynamic equilibrium and are free in a solution. Thus, single fatty chain surfactants generally exhibit significant detergency and are not suitable as carriers for hydrophobic compounds. In the biomedical field, micelles used as carriers must be biocompatible, i.e., the surfactant molecules forming the micelles can be biodegraded in vivo or harmlessly excluded from the body. Lipid molecules in nature have natural advantages in biocompatibility. The common lipoid molecules have double fatty chains, and lipophilic groups and hydrophilic groups of the lipoid molecules have similar volumes and are easy to form a vesicular structure. The lysophospholipid with a special structure contains lipophilic groups which are single-chain fatty acids, and the volume of the lipophilic groups is smaller than that of phosphorylcholine of a hydrophilic group, so that micelles are easy to form.

According to the principle of micelle formation, a stable micelle system can be obtained by increasing the association of the hydrophobic portion of the surfactant. In the prior art, the volume of a hydrophilic part is larger than that of a lipophilic part through the structural design of a linear amphiphilic block copolymer, so that a high-molecular micelle is formed. The stability of the micelle is improved by a large amount of oleophylic groups in the amphiphilic polymer, and the effective components are protected in the in vivo circulation. However, the size of the block copolymer polymer micelle is larger due to a large amount of oleophylic and hydrophilic groups, the hydration radius can reach more than 100 nm, and the tissue penetrability is reduced. Furthermore, the distribution and number of hydrophobic chains in the copolymer is difficult to control precisely, which affects the precise regulation of the kinetics of the release of the hydrophobic compound. In the field of biomedical applications, in particular, in the application of drug delivery and the encapsulation of hydrophobic compounds related to cosmetics, it is often necessary to flexibly adjust the size and stability of micelles according to the use conditions and the structure of the active ingredient, so that the micelles have high biocompatibility, tissue penetration, long-term stability and controllable release performance at the same time. In the preparation of surfactant molecules, the number and the structure of hydrophobic parts (namely oleophilic groups) and hydrophilic parts (namely hydrophilic groups) are accurately adjustable, and the method is a practical way for obtaining micelle type carriers with wide practicability.

Disclosure of Invention

The invention aims to prepare a stable polyethylene glycol derivative, a preparation method and application thereof, wherein the polyethylene glycol derivative can form micelles when dispersed in a solvent, and the micelles can be used for wrapping and delivering hydrophobic substances, so that long-acting release and high-efficiency tissue penetrating delivery of the hydrophobic substances are realized.

The invention realizes the aim through the following technical scheme:

a polyethylene glycol derivative comprising a group containing a fatty chain and polyethylene glycol, said group containing a fatty chain being linked to polyethylene glycol via a linking group, wherein at least one of said groups containing a fatty chain is a phospholipid group.

According to the invention, the structure of the polyethylene glycol derivative is PEG- (L)₁-R₁)_nWherein PEG is polyethylene glycol group, L₁Is a divalent linking group, n is an integer between 2 and 10, R₁Same or different, independently selected from the group consisting of phospholipid groups, C_4-40A hydrocarbyl group, and wherein at least one R is₁Are phospholipid groups.

According to the invention, the polyethylene glycol groups are selected from linear polyethylene glycol groups or multi-arm polyethylene glycol groups.

According to the invention, R₁The same or different, are independently selected from the group consisting of phospholipid groups,C_4-24Hydrocarbon radicals, e.g. phospholipid radicals, C_4-24Alkyl radical, C_4-24Alkenyl radical, C_4-24Alkynyl, and wherein at least one R₁Are phospholipid groups.

According to the present invention, the phospholipid group is preferably a structure represented by the following formula (I) or formula (II),

or

Wherein R is₂、R₃The same or different, are independently selected from C_4-40Hydrocarbyl represents a bond.

Preferably, R₂、R₃The same or different, are independently selected from C_4-24Hydrocarbyl radicals, e.g. C_4-24Alkyl radical, C_4-24Alkenyl radical, C_4-24Alkynyl.

According to the present invention, the number of the phospholipid groups in the polyethylene glycol derivative is 1 to 10.

According to the invention, the divalent linking group L₁Is capable of cleaving the radical R₁Any group attached to the polyethylene glycol group PEG, and a divalent linking group L₁The same or different, the divalent linking group L₁Contains the following groups: -CO-NH-, -CO-O-, -CS-O-, -NH-CO-NH-, -CO-NH-CO-, -O-, -S-S-, -NH-,an azido-alkynyl cycloaddition linking group, a tetrazinyl-alkenyl cycloaddition linking group.

The divalent linking group L₁For example, it may be: -CO-CH₂-CH₂-CO-NH-CH₂-CH₂-、-CO-CH₂-CH₂-CO-NH-、-CH₂-CO-NH-CH₂-CH₂-、-CH₂-CO-NH-、-S-S-、。

According to the invention, the divalent linking group L if the polyethylene glycol group is selected from linear polyethylene glycol groups₁Is equal to 2, said divalent linking group L if said polyethylene glycol group is selected from multi-armed polyethylene glycol groups₁The number of (a) is equal to the number of arms of the multi-arm polyethylene glycol.

According to the invention, the multi-armed polyethylene glycol group has a number of arms of 3, 4, 5, 6, 7, 8, 9 or 10, and the number of arms is the same as n. Illustratively, the multi-arm polyethylene glycol is at least one of a three-arm polyethylene glycol, a four-arm polyethylene glycol, a five-arm polyethylene glycol, a six-arm polyethylene glycol, a seven-arm polyethylene glycol, an eight-arm polyethylene glycol, a nine-arm polyethylene glycol, or a ten-arm polyethylene glycol.

According to the present invention, the multi-armed polyethylene glycol group comprises a polyethylene glycol repeat unit and a branched core structure selected from glycerol, glycidyl ether, ethylene glycol glycidyl ether, pentaerythritol, diglycidyl pentaerythritol ether, triglycidyl ether, gallic acid ether, or ethylene glycol gallic acid ether; wherein the glycidyl ether is diglycidyl ether, triglycidyl ether, tetraglycidyl ether, pentaglycidyl ether and hexaglycidyl ether.

According to the invention, the number average molecular weight of the polyethylene glycol group is 500-60000 Da, preferably 1000-50000Da, and more preferably 2000-40000 Da.

The invention also provides a polyethylene glycol derivative which is prepared by reacting the following raw materials:

(a) phospholipid compound, and (b) polyethylene glycol PEG- (Y) with Y as terminal group_nAnd optionally (c) a compound R containing an aliphatic chain₄-X₁Wherein (a) an amino group or a hydroxyl group in the phospholipid compound and (b) PEG- (Y)_nY in (b) is reacted with (c) R₄-X₁X in (1)₁And (b) PEG- (Y)_nReacting Y in the step (1);

wherein, PEG is linear polyethylene glycol group or multi-arm polyethylene glycol group, and n is an integer between 2 and 10;

the radicals Y, which are identical or different, are independently selected from the group consisting of-R₅-a, said a being a reactive group: -COOH, amino, -OH, -SH, -C.ident.N, -C.ident.C, -C = C,N-Hydroxyl succinimide ester group, maleimide group, dithiopyridyl group, vinyl sulfone group, isocyanate group and tetrazine group; the R is₅In the absence or divalent linking groups, e.g. R₅is-CO-C_1-6Alkyl-, -C_1-6Alkyl-, -NH-C_1-6Alkyl-;

the group X₁The same or different, are independently selected from the following reactive groups: -COOH, amino, -OH, -SH, -C.ident.N, -C.ident.C, -C = C,N-Any one of a hydroxysuccinimide ester group, a maleimide group, a dithiopyridyl group, a vinylsulfone group, an isocyanate group and a tetrazine group;

R₄is C_4-40A hydrocarbyl group.

According to the present invention, the raw material (a) is one or more kinds of phospholipid compounds. The phospholipid compound is lipid containing phosphoric acid and amino or hydroxyl. The phospholipid compound has an amino group or a hydroxyl group which reacts with the reactive group in the raw material (b).

According to the present invention, the phospholipid compound may be natural or synthetic.

Illustratively, the phospholipid compound is, for example, a glycerophospholipid (also referred to as phospholipid), such as Phosphatidylethanolamine (PE), Phosphatidylserine (PS), Phosphatidylglycerol (PG), or Phosphatidic Acid (PA), a lysophospholipid, such as lysophosphatidylcholine (L PC), lysophosphatidylethanolamine (L PE), lysophosphatidylinositol (L PI), or lysophosphatidic acid (L PA), a lysophospholipid, a sphingomyelin, or the like.

Illustratively, the phosphatidylethanolamine is selected from the group consisting of dihexanoylphosphatidylethanolamine, dioctanoylphosphatidylethanolamine, didecanoylphosphatidylethanolamine, dilauroylphosphatidylethanolamine (D L PE), Dimyristoylphosphatidylethanolamine (DMPE), Dipalmitoylphosphatidylethanolamine (DPPE), Distearoylphosphatidylethanolamine (DSPE), Dioleoylphosphatidylethanolamine (DOPE), Dicapryoylphosphatidylethanolamine (DEPE), 1, 2-Docosahexaenoyl (DHA) phosphatidylethanolamine, 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), 1-stearoyl-2-oleoylphosphatidylethanolamine (SOPE), and the lysophosphatidylethanolamine is selected from the group consisting of myristoyl lysophosphatidylethanolamine (L MPE), palmitoyl lysophosphatidylethanolamine (L PPE), and stearoyl lysophosphatidylethanolamine (L SPE).

According to the invention, in the raw material (b), Y is a capping group of polyethylene glycol, and the raw material (b) is polyethylene glycol with various capping groups which exist in the prior art and can be purchased or synthesized autonomously.

According to the present invention, the raw material (b) may be a linear polyethylene glycol having a terminal group substituted.

According to the present invention, the raw material (b) may be a multi-arm polyethylene glycol with a substituted terminal group, illustratively, at least one of a three-arm polyethylene glycol, a four-arm polyethylene glycol, a five-arm polyethylene glycol, a six-arm polyethylene glycol, a seven-arm polyethylene glycol, an eight-arm polyethylene glycol, a nine-arm polyethylene glycol, or a ten-arm polyethylene glycol.

According to the invention, the polyethylene glycol has a number average molecular weight of 500-.

According to the invention, the starting material (c) may or may not be present. The group X in the starting material (c)₁A reactive group which can react with the reactive group of the group Y in the starting material (b) to form a group R₄Linked together with polyethylene glycol.

The polyethylene glycol derivative has the structure of PEG- (L)₁-R₁)_nWherein PEG is polyethylene glycol group, L₁Is a linking group, n is an integer between 2 and 10, R₁Same or different, independently selected from the group consisting of phospholipid groups, C_4-40A hydrocarbyl group, and wherein at least one R is₁Are phospholipid groups.

The invention also provides a preparation method of the polyethylene glycol derivative, which comprises the following steps:

the phospholipid compound (a) and the polyethylene glycol PEG- (Y) with Y as the end group (b)_nAnd optionally (c) a compound R containing an aliphatic chain₄-X₁Mixing and reacting to obtain the polyethylene glycol derivative.

According to the invention, the above-mentioned starting materials are mixed in a solvent, for example selected from N, N-dimethylformamide.

According to the invention, the method comprises the following steps:

(1) mixing (a) phospholipid compound and optionally (c) aliphatic chain-containing compound R₄-X₁Dissolving in a solvent to obtain a mixed solution A;

(2) polyethylene glycol PEG- (Y) with the end group of (b) being Y_nDissolving in a solvent to obtain a mixed solution B;

(3) and mixing the mixed solution A and the mixed solution B, and reacting at room temperature to prepare the polyethylene glycol derivative.

According to the invention, the method may further comprise the steps of:

(1') dissolving the phospholipid compound (a) in a solvent to obtain a mixed solution A;

(2') adding (b) polyethylene glycol PEG- (Y) with Y as the end group_nDissolving in a solvent to obtain a mixed solution B;

(3') mixing the mixed solution A and the mixed solution B, reacting at room temperature, and optionally adding (c) a compound R having an aliphatic chain₄-X₁And preparing the polyethylene glycol derivative.

According to the present invention, step (3) or step (3') further comprises adding a terminating reagent during the reaction to terminate the reaction. The terminating agent is selected, for example, from water.

According to the present invention, when the raw material (c) is not present, the phospholipid compound as the raw material (a) and PEG- (Y) as the raw material (b)_nThe molar ratio of (a) to (b) is n: 1.

According to the bookIn the present invention, when the raw material (c) is present, the phospholipid compound as the raw material (a) and the R as the raw material (c)₄The sum of the moles of-X and the starting material (b) PEG- (Y)_nThe molar ratio of (a) to (b) is n: 1.

Wherein the phospholipid compound as the raw material (a) and the R as the raw material (c) are₄The molar ratio of-X is not particularly limited, and it is sufficient that the polyethylene glycol derivative obtained by the preparation contains at least one phospholipid group.

The invention also provides application of the polyethylene glycol derivative in preparation of micelles.

The invention also provides a micelle, which comprises the polyethylene glycol derivative.

According to the present invention, the micelle is formed by dissolving the polyethylene glycol derivative into a solvent. Preferably, the polyethylene glycol derivative is dissolved in an aqueous solution so that the concentration thereof is 1 to 1000 times the critical micelle concentration.

According to the invention, the micelle is composed of one polyethylene glycol derivative or is formed by combining more than two polyethylene glycol derivatives. The combination may be a combination of binary, ternary or polyvalent polyethylene glycol derivatives, and the mass percentage of each polyethylene glycol derivative in the composition is not particularly limited as long as the composition is capable of forming micelles.

The invention also provides a preparation method of the micelle, which comprises the following steps:

dissolving the polyethylene glycol derivative in a solvent to make the concentration of the polyethylene glycol derivative be 1-1000 times of the critical micelle concentration.

According to the present invention, the concentration is preferably 1 to 200 times the critical micelle concentration, more preferably 1 to 50 times the critical micelle concentration.

According to the invention, the solvent is, for example, an aqueous solution.

According to the present invention, the critical micelle concentration of the polyethylene glycol derivative is measured by a method known in the art, and includes, for example, a conductance method, a surface tension method, a drop volume method, an ultrafiltration curve method, a single-point ultrafiltration method, a two-point ultrafiltration method, an ultraviolet spectrophotometry method, a dye adsorption method, a light scattering luminescence method, a fluorescence probe method, a solubility method.

The invention also provides the use of the micelle in the coating and delivery of hydrophobic compounds.

According to the invention, the hydrophobic compound is chosen, for example, from hydrophobic drugs, such as paclitaxel, hydrophobic cosmetics, such as squalane, tocopherols.

The present invention also provides a delivery system comprising a micelle and a hydrophobic compound, the hydrophobic compound being located inside the micelle.

According to the invention, the delivery system is prepared by contacting the solution containing the micelles with a hydrophobic compound, optionally evaporating the solvent.

According to the invention, the molar ratio of the hydrophobic compound to the micelles is 0.0001 to 1000, preferably 0.001 to 100, more preferably 0.01 to 50, for example 0.005:1, 0.01:1, 0.02:1, 0.05:1, 0.1:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.8:1, 1:1, 2:1, 3:1, 4:1, 5:1, 8:1, 10:1, 12:1, 15:1, 18:1, 20:1, 25:1, 28:1, 30:1, 35:1, 40:1, 45:1, 50:1, 80:1, 100:1, 120:1, 150:1, 180:1, 200:1, 300:1, 400:1, 500:1, 600:1, 700:1, 800:1, 900:1, 1000: 1.

Wherein, mechanical stirring, shaking or ultrasonic dispersion is used in the contact process.

Wherein the solution containing the micelles may be, for example, an organic solution containing micelles, and the solvent of the organic solution is not particularly limited as long as it can dissolve the lipid molecules, and may be, alternatively, one of alcohol, ether, ketone, ester, amide, sulfoxide, alkane, cycloalkane, aromatic hydrocarbon, chloroalkane, or a mixture thereof.

The invention has the beneficial effects that:

(1) the invention provides a polyethylene glycol derivative, which contains a phospholipid structure and a plurality of fatty chains, wherein the volume and the association strength of a hydrophobic part (namely a lipophilic group) can be adjusted by changing the chain length, the saturation and the number of the fatty chains, while the volume of a hydrophilic part can be adjusted by selecting the chain length and the number of branches of a polyethylene glycol macromolecule, and the two are cooperated to obtain stable micelles with different micelle sizes.

(2) Particularly, the polyethylene glycol derivative realizes the stable existence of the small-particle-size nano micelle in an organic solvent, and overcomes the technical problem of low tolerance of the liposome and the single-fatty-chain micelle to the organic solvent under the conventional condition.

(3) According to the invention, a proper polyethylene glycol derivative can be selected to form a micelle according to the structure of the active ingredient and the use condition required in the biomedical field, so that the size and the stability of the micelle are flexibly adjusted through a 'bottom-up' strategy, a nano-encapsulation and delivery system is efficiently constructed, and the efficient encapsulation, delivery and release of the hydrophobic active ingredient by the micelle are realized.

(4) The polyethylene glycol derivative micelle has high tissue penetrability, long-acting stability and controllable release performance. Particularly, the polyethylene glycol derivative of the invention contains a phospholipid structure, has outstanding biocompatibility, and is suitable for the field of biological medicines with high requirements on biological safety.

[ terms and explanations ]

The "alkyl" in the invention represents saturated or unsaturated aliphatic hydrocarbon with 4-40 carbon atoms, such as 8-40 carbon atoms, the unsaturated aliphatic hydrocarbon contains unsaturated groups, optionally alkenyl or alkynyl, and the unsaturated groups can be one or more than two. Preferably, the hydrocarbyl group is "C_4-24Hydrocarbyl groups "are for example C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24.

The "alkyl" as referred to herein represents a straight-chain, branched-chain alkyl group having 4 to 40, such as 8 to 40, carbon atoms, and preferably the alkyl group is "C_4-24Alkyl groups "; for example, octyl, decyl, undecyl, dodecyl, tetradecyl, hexadecyl, octadecyl.

The "alkenyl" group in the present invention represents a straight-chain or branched alkenyl group having 4 to 40 carbon atoms, for example, 8 to 40 carbon atoms, and preferably, the number of double bonds is an integer of 1 to 6. Preferably, the alkene isRadical being "C_4-24Alkenyl radicals "; for example, octenyl, decenyl, undecenyl, dodecenyl, tetradecenyl, hexadecenyl, octadecenyl.

The "alkynyl" in the present invention represents a straight-chain, branched-chain alkynyl group having 4 to 40 carbon atoms, and preferably, the number of triple bonds is an integer of 1 to 6. Preferably, said alkynyl is "C_4-24Alkynyl "; for example, octynyl, decynyl, undecylynyl, dodecynyl, tetradecynyl, hexadecylynyl, octadecynyl.

Said "amino" according to the invention represents the group-NH₂、-NHR⁷Wherein R is⁷Independently selected from H, alkyl, aryl, heteroaryl, heterocyclic radical.

The ether group of the invention represents a group-OR⁸Wherein R is⁸Independently selected from C_1-6Alkyl, - (CH)₂-CH₂O)n-CH₂-CH₃(n is greater than 2); examples of the ether group include methyl ether, ethyl ether, propyl ether, isopropyl ether, butyl ether, isobutyl ether, tert-butyl ether, polyoxyethylene ether group having an ethylene oxide number of 9 to 12, and the like.

Suitable reactive groups are well known in the art and may be, for example, hydroxyl, amino, carboxyl, aldehyde, ketone, ester, thiol, maleimide, α -halocarbonyl, alkynyl, alkenyl, azido, tetrazinyl.

The "linking group" as used herein means a group which links two groups, specifically, a phospholipid compound or a hydrocarbon compound and the polyethylene glycol.

The "end group substituted polyethylene glycol" refers to that the hydroxyl group of the end capping group of the polyethylene glycol is substituted by other groups. Various end-substituted polyethylene glycols are known in the art, for example, polyethylene glycols having the following end-capping groups: methoxy, ethoxy, propoxy, amino, carboxyl, alkynyl, azido, and the like.

The linear polyethylene glycol refers to the linear polyethylene glycol molecular chain, namely the linear polyethylene glycol contains a linear vinyl ether repeating unit and two hydroxyl end-capping groups.

The "multi-arm polyethylene glycol," also referred to as "multi-arm PEG polyol," as described herein, refers to a branched poly (ethylene glycol) having 3 to 10 branches ("arms") with a hydroxyl end-cap. Suitable multi-arm polyethylene glycols include, but are not limited to, dendritic, comb, and star poly (ethylene glycol). Typically, useful multi-arm polyethylene glycols have a molecular weight of from about 450 to about 200000 daltons, or from about 2000 to about 40000 daltons. It should be recognized that multi-arm polyethylene glycols are generally heterogeneous mixtures of species having a distribution of arm lengths and, in some cases, different arm numbers. When the multi-arm polyethylene glycol has a distribution of species with different arm numbers, it can be expressed on the basis of the average arm number in the distribution. For example, in one embodiment, the multi-arm polyethylene glycol can be an 8-arm star PEG polyol comprising a mixture of multi-arm star PEG polyols, some of which have fewer than 8 arms, some of which have more than 8 arms; however, the multi-arm star PEG polyol in this mixture has an average of 8 arms. Mixtures of multi-arm PEG polyols having different arm numbers and/or different molecular weights may additionally be used as starting materials.

The "Phospholipid" (Phospholipid), the "Phospholipid compound" and the "Phospholipid" as used herein refer to a lipid containing a phosphoric acid, and belong to complex lipids. The main components are glycerophospholipids and sphingomyelin, and the phospholipids composed of glycerol are called glycerophospholipids; phospholipids, consisting of sphingosine, are called sphingomyelins. The structure is characterized in that: having a hydrophilic head (hydrophic head) consisting of a phosphate-linked substituent (containing an ammonia base or alcohol) and a hydrophobic tail (hydrophthalil) consisting of a fatty acid chain.

The glycerophospholipid is also called phosphoglyceride, the main chain is glycerol-3-phosphoric acid, the other two hydroxyl groups in glycerol molecules are esterified by fatty acid, and phosphate groups can be esterified by micromolecular compounds with different structures to form various phosphoglycerides. There may be several per phospholipid depending on the constituent fatty acids.

The "lysophospholipid" (lysolecithin) according to the present invention is a kind of phospholipid, hydrolyzed phospholipid, which is a compound produced by hydrolyzing a phospholipid with phospholipase a1, phospholipase a2, or phospholipase B, etc. lysophospholipids are classified into lysophosphatidylcholine (L PC), lysophosphatidylethanolamine (L PE), lysophosphatidylinositol (L PI), lysophosphatidic acid (L PA), a compound in which an acyl group at the hydroxyl group at the 1-or 2-position of the glycerol backbone is combined into an ester (a decomposition product of phospholipase a2 or a decomposition product of a 1), depending on the substrate source.

The "Sphingomyelin" (Sphingomyelin) in the present invention refers to sphingosine or dihydrosphingosine-containing phospholipids, which do not contain glycerol in the molecule, and are molecules of fatty acid linked to the amino group of sphingosine via amide bond. Sphingosine or dihydrosphingosine is an amino diol with a long aliphatic chain. Has a hydrophobic long-chain aliphatic tail and two polar heads with hydroxyl and amino groups.

Drawings

Fig. 1 is a schematic view of the structure of the micelle formed by the polyethylene glycol derivative and the polyethylene glycol derivative according to the present invention.

FIG. 2 is a mass spectrum of the molecular weight of the 1, 2-distearoyl-3-phosphatidylethanolamine-n-octadecanethiol-polyethylene glycol derivative synthesized in example 1.

FIG. 3 is a fluorescence emission spectrum (A) of Critical Micelle Concentration (CMC) of micelle determined by the fluorescence probe method and a concentration titration curve (B) of CMC determined by the fluorescence probe method in example 14, wherein I₁And I₃The intensity is deconvoluted and corrected, and the influence of the long-wave band emission peak is removed.

FIG. 4 is a graph showing the determination of micelle hydration radius by dynamic laser light scattering in example 15.

FIG. 5 is a graph showing the effect of micelle encapsulation of pyrene molecules by lipid molecules of a multi-fatty-chain multi-arm polyethylene glycol derivative in absorbance detection of example 16.

Detailed Description

The preparation method of the present invention will be described in further detail with reference to specific examples. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

The molecular weight of diisopropylethylamine used in the following examples was 129 Da, and the manufacturer was Beijing chemical plant. Used in the following examplesN,NThe-dimethylformamide is anhydrous DMF, the purity is analytical purity, and the manufacturer is a Beijing chemical plant.

The experimental methods used in the following examples are all conventional methods unless otherwise specified; reagents, materials and the like used in the following examples are commercially available unless otherwise specified. Optionally indicating the presence or absence of the stated feature, and also indicating that the stated feature must be present, although the particular choice may be arbitrary.

The number of polyethylene glycol repeating units in the following examples is schematically shown as an average.

1, 2-Distearoyl-3-phosphatidylethanolamine (DSPE, molecular weight 748 Da) used in the examples described below was obtained from Mirida technologies, Inc. of Beijing, and 1, 2-dilauroyl-phosphatidylethanolamine (D L PE, molecular weight 579 Da) used in the examples was obtained from Xinqiao, Hangzhou, Biotech, Inc.

Octadecylamine (molecular weight 270 Da) used in the following examples was purchased from Shanghai Xiandong Biotechnology Ltd. N-dodecylamine (molecular weight 185 Da) was used from Shanghai Allantin Biotech Co., Ltd.