CN108187059B - Stimulus-response type amphiphilic cyclodextrin polymer carrier, preparation and application thereof in preparation of sustained and controlled release drugs - Google Patents
Stimulus-response type amphiphilic cyclodextrin polymer carrier, preparation and application thereof in preparation of sustained and controlled release drugs Download PDFInfo
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Abstract
The invention belongs to the field of pharmaceutical preparations, and particularly discloses an amphiphilic cyclodextrin polymer drug carrier, which is characterized in that: taking cyclodextrin as a core, wherein a hydrophilic chain segment is grafted on the 6-position modification of at least one sugar ring of the cyclodextrin core; the 2-position and/or 3-position of at least one sugar ring is modified with a hydrophobic chain segment. And the chemical bond with stimulation responsiveness to the tumor microenvironment is connected with the hydrophilic end and the cyclodextrin, so as to achieve the purpose of directional release. In addition, the invention also discloses a preparation method and application of the carrier. The amphiphilic cyclodextrin drug-loaded system prepared by the method has the advantages of proper particle size, uniform shape, large drug-loaded amount, good stimulation responsiveness and high sustained and controlled release performance.
Description
The technical field is as follows:
the invention belongs to the field of pharmaceutical preparations; in particular to a drug carrier with hydrophilic and hydrophobic properties.
Background
In recent years, the incidence of cancer has been on the rise with environmental deterioration, population growth, aging, and lifestyle changes. According to data of 'annual report on Chinese tumor registration' in 2017, now, on average, 7 people are diagnosed with cancer every minute every day, 10000 people are diagnosed every day, and the probability of suffering from cancer in the life of the people is 36%. Most of the anticancer drugs applied clinically at present are chemotherapeutic drugs, which can kill cancer cells and greatly damage normal cells, resulting in low drug effect and serious side effect. Therefore, the invention of a drug which specifically kills cancer cells, has low toxic and side effects and can be slowly released is a problem to be solved.
In order to solve the problem of large side effect of anticancer drugs, some methods are reported in the prior art, for example, chinese patent publication No. CN103396521A discloses an amphiphilic β -CD star polymer having a hydrophobic block and a hydrophilic block, which is characterized in that: the amphiphilic beta-CD star polymer takes beta-CD as a core, and an amphiphilic polymer is connected to 3-6 hydroxyl groups of the core to form a three-arm, four-arm, five-arm or six-arm amphiphilic beta-CD star polymer; the hydrophobic block of the attached amphiphilic polymer is polymerized from Acrylic Acid (AA) and Methyl Methacrylate (MMA), and the hydrophilic block is polymerized from N-vinyl pyrrolidone (PNVP). The polymer is pH sensitive, can stimulate response type to release hydrophobic drugs, and has the maximum drug loading rate of 21.44%. Acrylic and methacrylic polymers as hydrophobic end, acrylic and methacrylic polymers are non-toxic, but the monomers are toxic, which limits the application of this polymer on the one hand; and the drug loading is not large, resulting in more carriers being required and resulting in increased potential toxicity.
For another example, chinese patent publication No. CN101284885A discloses an amphiphilic cyclodextrin-based polymer and a preparation method thereof, having a hydrophilic end block and a hydrophobic end block, characterized in that: the hydrophilic end of the prepared amphiphilic cyclodextrin polymer is polyethylene glycol (PEG), the hydrophobic end of the amphiphilic cyclodextrin polymer is degradable Polylactide (PCL), and the polyethylene glycol (PEG) and the degradable Polylactide (PCL) are connected with Cyclodextrin (CD) to form an amphiphilic cyclodextrin drug-loading system (CD-PCL-PEG), so that the entrapment of a hydrophobic anticancer drug adriamycin is realized, the drug toxicity is expected to be covered, the toxic and side effects are reduced, and the maximum drug-loading amount is 20.1%. The amphiphilic cyclodextrin polymer prepared by the method cannot stimulate response type release of drugs.
Although the amphiphilic cyclodextrin is successfully prepared by the existing method and the toxicity of the anticancer drug is successfully covered, the acrylic acid monomer is toxic, and the polyacrylic acid prepared by the existing method is non-toxic, but cannot ensure that all acrylic acid is successfully polymerized, so that the potential toxicity exists; in addition, there is a problem of low drug loading; the prior art needs a carrier and a preparation which have good stimulation responsiveness, high drug loading, no toxicity and good sustained and controlled release performance.
Disclosure of Invention
The first purpose of the invention is to provide an amphiphilic cyclodextrin polymer drug carrier (also referred to as a carrier for short in the invention), aiming at improving the drug loading rate of the carrier.
The second purpose of the invention is to provide a preparation method of the amphiphilic cyclodextrin polymer drug carrier.
The third objective of the invention is to provide an application of the amphiphilic cyclodextrin polymer drug carrier, and the amphiphilic cyclodextrin polymer drug carrier and a hydrophobic drug are mixed in water for self-assembly (also referred to as self-polymerization in the invention) so as to obtain a drug preparation with a sustained and controlled release effect.
The fourth purpose of the invention is to provide a sustained-release preparation (also called drug delivery system) assembled by the amphiphilic cyclodextrin polymer drug carrier.
The amphiphilic cyclodextrin polymer drug carrier takes cyclodextrin as a core, wherein a hydrophilic chain segment is grafted on the 6-site modification of at least one sugar ring of the cyclodextrin core; the 2-position and/or 3-position of at least one sugar ring is modified with a hydrophobic chain segment.
According to the invention, cyclodextrin is innovatively used as a modification core, a hydrophilic chain segment is modified at the 6-position of the cyclodextrin, and a hydrophobic chain segment is modified at the 2-position and/or the 3-position of the cyclodextrin, so that the performance of the obtained carrier can be unexpectedly improved, and the drug loading capacity of the carrier is improved. Meanwhile, taking the application of the self-assembly amphiphilic cyclodextrin in tumor drugs as an example, the particle size of the self-assembly amphiphilic cyclodextrin can be controlled to be 40-100 nm by adjusting the lengths of a hydrophilic chain and a hydrophobic chain, so that the self-assembly amphiphilic cyclodextrin is beneficial to aggregating a drug-loading system at a tumor position through an EPR effect (namely the high-permeability and long-retention effects of solid tumors), the side effect is reduced, and the curative effect is improved; the drug-carrying system is connected through a stimulus-responsive chemical bond, and can be specifically released in tumor tissues, so that side effects are further reduced.
The invention provides a novel amphiphilic cyclodextrin carrier and a preparation method thereof, the amphiphilic cyclodextrin carrier prepared by the method can greatly improve the drug-loading rate, has proper size, can realize irritant release to the reducing environment in the microenvironment of tumor tissues, greatly reduces the toxic and side effects of the amphiphilic cyclodextrin carrier, and increases the curative effect.
The cyclodextrin is preferably alpha-, gamma-or beta-cyclodextrin; beta-cyclodextrin is particularly preferred.
The 6-position of the sugar ring can be modified with a hydrophilic segment. Further research shows that the improvement of the number of the 6-position modified hydrophilic chain segments of the sugar ring helps to further improve the performance of the carrier.
Preferably, the method comprises the following steps: the 6-position of each sugar ring of the cyclodextrin is modified with a hydrophilic chain segment.
In the present invention, the hydrophilic segment is preferably a polymer segment having good hydrophilicity.
Further research shows that the hydrophilic chain segment is polyethylene glycol hydrophilic chain segment. The hydrophilic end of the polyethylene glycol can improve the water solubility of the drug, is beneficial to avoiding the recognition and the damage of an immune system in a drug-carrying system, can effectively prolong the circulation time of the drug-carrying system in vivo and improves the bioavailability.
In the invention, the structural formula of the polyethylene glycol hydrophilic chain segment is-PEG-O-CH3。
More preferably, the molecular weight of the polyethylene glycol hydrophilic segment is 200 to 8000.
Preferably, the method comprises the following steps: the hydrophilic chain segment is connected with the 6-position of the cyclodextrin through a stimulus response type chemical bond; the stimulus-responsive chemical bond is one or more of a pH stimulus-responsive chemical bond, a temperature stimulus-responsive chemical bond or an oxidation-reduction stimulus-responsive chemical bond.
Further preferably, the redox stimulus-responsive chemical bond is a disulfide bond. That is, the hydrophilic segment is attached to the primary carbon at the 6-position of the cyclodextrin sugar ring by a disulfide bond.
The inventor researches and discovers that increasing the number of the hydrophobic chain segments at the 2-position and the 3-position of the sugar ring is helpful to further cooperate with the hydrophilic chain segments and further improve the effect of the obtained carrier, for example, the drug loading rate and the sustained and controlled release performance of the obtained carrier are obviously improved.
More preferably, the 2-position and/or 3-position of each sugar ring of the cyclodextrin is modified with a hydrophobic segment. That is, the 2-position and 3-position of 7 sugar rings of the cyclodextrin, and 14 reaction sites in total are modified with hydrophobic segments.
More preferably, in the amphiphilic cyclodextrin polymer drug carrier, a hydrophilic chain segment is modified at the 6-position of each sugar ring; the 2-position and the 3-position on each sugar ring are both modified with hydrophobic chain segments. In this way, it is helpful to synergistically improve the performance of the resulting carrier by the hydrophilic coordination of the 6-position and the hydrophobic coordination of the 2, 3-positions.
The hydrophobic segment may be selected from groups having hydrophobicity, which are well known to those skilled in the art.
In the present invention, the hydrophobic segment is preferably modified at the secondary hydroxyl group at the 2-position and/or 3-position; that is, a hydrophobic segment is used in place of H on the secondary hydroxyl group at the 2-position and/or 3-position.
Preferably, the method comprises the following steps: the hydrophobic chain segment is a C1-C20 alkyl chain segment; or a C1-C20 hydrocarbyl segment substituted by at least one substituent selected from halogen, C1-C6 alkyl, C1-C6 alkoxy and benzyl. The hydrocarbon chain segment of C1-C20 is the alkyl group with carbon number, or the alkylene group or the alkyne group containing unsaturated bonds.
More preferably, the hydrophobic chain segment is a linear or branched saturated alkyl chain segment of C5-C20; further preferred are straight-chain saturated hydrocarbon segments. It has been found that the hydrophobic segment of the preferred segment and the hydrophilic segment of the present invention cooperate to assist in synergistically increasing the drug loading.
Further preferably, the hydrophobic segment is dodecyl. Further preferred hydrophobic segments may cooperate with the hydrophilic segments of the present invention to synergistically increase drug loading to 40%.
In the invention, in a most preferable amphiphilic cyclodextrin polymer drug carrier, 6-position on each sugar ring is modified with PEG hydrophilic chain segment; the 2-position and the 3-position on each sugar ring are modified with n-dodecyl (the structural formula is shown as formula 1, the structure is a bouquet type conglomerate structure, and the hydrophilic chain segment and the hydrophobic chain segment are not all shown in the formula 1). Thus, the combination of the hydrophilicity of the 6-position and the hydrophobicity of the 2-position and the 3-position is facilitated, the performance of the obtained carrier is improved synergistically, and the drug loading rate of the carrier is obviously improved.
The invention also provides a preparation method of the amphiphilic cyclodextrin polymer drug carrier, which comprises the following steps:
step (a): protection of the 6-hydroxy group of cyclodextrins
Protecting 6-primary alcohol of cyclodextrin to obtain 6-hydroxyl protected cyclodextrin a;
step (b): hydrophobic modification of hydroxyl groups at position 2 and/or 3 of the cyclodextrin;
reacting a with a source material of a hydrophobic chain segment under alkali to obtain a 2-position and/or 3-position hydroxyl hydrophobicity modified product b;
step (c): deprotection of the hydroxyl group at position 6 of product b
Hydrolyzing the b under an acidic condition, and removing the primary hydroxyl protecting group at the 6 th position to obtain a product c;
(d) and the product c is directly connected with the hydrophilic chain segment or connected through a stimulus response type chemical bond to prepare the carrier.
In the invention, through the preparation method, hydrophobic groups are modified at 2 and 3 positions under the condition of 6-position protection in advance, and then a hydrophilic chain segment is connected at 6 positions, so that a carrier with good performance can be prepared.
Researches show that the amphiphilic cyclodextrin synthesized by the preparation method can adjust the drug-loading rate and the sustained and controlled release effect of a drug carrier by adjusting the hydrophobic chain. It was found that if the hydrophobic chain is short (C0-C4) or the number of linkages is small, the drug loading is greatly affected.
In the invention, a proper primary hydroxyl protecting group is selected to match with the preparation steps, so that the steric hindrance can be reduced as much as possible while the carrier can selectively react with the 2, 3-primary hydroxyl, the purpose of complete substitution is achieved, the completely substituted hydrophobic chain segment is matched with the hydrophilic chain segment, the carrier can load a medicament through simple self-polymerization, and the medicament loading capacity can be obviously improved in a synergistic manner.
Preferably, in the step (a), the primary hydroxyl protecting group, cyclodextrin and triethylamine are reacted in an organic solvent at 20-80 ℃ for 72-120 h to protect the cyclodextrin 6-primary alcohol, then the solvent is removed, and the product is purified by silica gel column chromatography or recrystallization, and dried to obtain the 6-hydroxyl protected cyclodextrin.
Preferably, the material source of the primary hydroxyl protecting group is one or more of triphenylchloromethane, tert-butyldimethylchlorosilane and p-toluenesulfonyl chloride. The inventor researches and discovers that the primary hydroxyl can be controlled to be completely substituted or partially substituted by the protective material.
In the invention, in the step (a), the molar ratio of the cyclodextrin to the primary hydroxyl protecting group is 1: 3-1: 14.
In the present invention, in step (b), the base may be any base known to those skilled in the art that can alkylate a secondary alcohol; in the present invention, sodium hydride is preferred.
The source material of the hydrophobic chain segment can be 1-brominated C1-C20 alkyl; preferably 1-brominated C5-C20 hydrocarbyl; further preferred is 1-bromododecane.
The inventors have surprisingly found that in step (b), in addition to the kind of good synergistic alkylating agent (source material of the hydrophobic segment), the need to add both base and hydrophobic segment in portions helps to solve the technical problem of difficult complete long-chain alkylation which is difficult to solve in the art, and helps the hydrophobic segment to fully substitute secondary hydroxyl groups.
The batch adding method comprises the steps of dividing sodium hydride and bromoalkane into 2-3 parts respectively, firstly adding one part of sodium hydride, reacting for more than 4 hours, then adding one part of bromoalkane, reacting for 2-4 days, and repeating the steps until the reaction is finished.
The invention also provides: the preparation method of the carrier comprises the following steps:
(1) protection of the 6-hydroxy group of cyclodextrins
Reacting a primary hydroxyl protecting group, cyclodextrin and triethylamine in an organic solvent at 20-80 ℃ for 72-120 hours to protect 6-primary alcohol of cyclodextrin, removing the solvent, purifying the product by a silica gel column chromatography or recrystallization method, and drying to obtain 6-hydroxyl protected cyclodextrin a;
(2) alkylation of 2, 3-hydroxy groups of cyclodextrins
Fully reacting a with sodium hydride in an organic solvent, adding bromoalkane, reacting for 2-5 days at 0-50 ℃, quenching, removing the solvent, purifying the product by a gel chromatography separation method, and drying to obtain b;
(3) deprotection of the 6-hydroxy group of cyclodextrins
Hydrolyzing the b under an acidic condition, removing a primary hydroxyl protecting group, neutralizing unreacted acid with ammonia water, removing the solvent to obtain a crude product, and drying in vacuum to obtain a product c;
(4) iodination of 6-hydroxy cyclodextrin
c, reacting with triphenyl phosphorus and iodine simple substances in an anhydrous organic solvent at 60-90 ℃ for 18-36 h, reducing half of the solvent, adjusting the pH value to 9-10 by using sodium methoxide, precipitating by using a large amount of methanol, washing by using methanol for three times to obtain a product, and drying to obtain d;
(5) thiolation of the 6-hydroxy group of cyclodextrin
d, reacting the raw materials with thiourea in an organic solvent at the temperature of 60-100 ℃ for 16-28 h, removing the organic solvent, adding a proper amount of aqueous solution of alkali, performing reflux reaction for 0.5-1.5 h, cooling the reaction solution, adjusting the pH value to be less than 7, extracting with dichloromethane, washing with saturated salt water, removing the solvent, and obtaining a product e.
(6) Bromination of polyethylene glycol monomethyl ether
Reacting polyethylene glycol monomethyl ether with phosphorus tribromide in an anhydrous organic solvent at 0-30 ℃ for 18-28 h, slowly dropwise adding the reaction solution into water, adding dichloromethane for extraction, washing with saturated saline water to neutrality, drying, and removing the dichloromethane solvent by using a rotary evaporator to obtain bromo-polyethylene glycol monomethyl ether f.
(7) Thiolation of polyethylene glycol monomethyl ether
f, reacting the mixture with thiourea in an anhydrous organic solvent at 60-100 ℃ for 16-28 h, removing the organic solvent, adding a proper amount of alkali aqueous solution, carrying out reflux reaction for 0.5-1.5 h, cooling the reaction solution, adjusting the pH to be less than 7, extracting with dichloromethane, washing with saturated salt water, removing the solvent, and obtaining thiolated polyethylene glycol monomethyl ether g.
(8) Synthesis of amphiphilic cyclodextrin bouquet type polymer drug carrier
g. e fully reacting in an oxidizing environment to obtain the final amphiphilic cyclodextrin h.
According to the invention, by the preferred preparation method, the technical problem of low drug loading rate caused by mismatching of the hydrophobic chain segment and the hydrophilic chain segment can be solved, complete modification of the required chain segment is realized, the hydrophilicity is matched with the hydrophobicity, and the drug loading rate of the drug is improved.
Preferably, in step (1), the primary hydroxyl protecting group is provided by one or more of triphenylchloromethane, tert-butyldimethylchlorosilane and p-toluenesulfonyl chloride. In the invention, a proper primary hydroxyl protecting group is selected, so that the steric hindrance can be reduced as much as possible while the primary hydroxyl protecting group can selectively react with the primary hydroxyl, and the primary hydroxyl can be completely substituted; or increase the steric hindrance so that it does not completely substitute for the primary hydroxyl group. That is, the choice of protecting group plays a key role in controlling the complete or partial substitution of the primary hydroxyl group.
The molar ratio of the cyclodextrin to the primary hydroxyl protecting group is 1: 3-1: 14.
Preferably, in step (2), the alkyl bromide has the chemical formula R1-Br. Said R1Is C1-12 alkyl; more preferably a C5-12 linear alkyl hydrocarbon group. Preferred R1The hydrophilic chain segment is beneficial to cooperating with the hydrophilic chain segment to improve the drug loading rate.
a, bromoalkane and sodium hydride in a molar ratio of 1:7 to 1:60 (i.e. 1:7 to 60). In the reaction, bromoalkane and sodium hydride need to be added in batches, so as to achieve the purpose of fully reacting bromoalkane and cyclodextrin. The extended reaction time facilitates the total substitution of the secondary hydroxyl groups of the cyclodextrin. The proportion of the three, the reaction time, the feeding method and the selection of bromoalkane jointly determine the substitution degree of the secondary hydroxyl.
The batch adding method comprises the steps of dividing sodium hydride and bromoalkane into 2-3 parts (for example, dividing the sodium hydride and the bromoalkane into equal parts), firstly adding one part of sodium hydride, reacting for more than 4 hours, then adding one part of bromoalkane, reacting for 2-4 days, and repeating the steps until the reaction is finished. For example, after a and 1/3 parts of sodium hydride for more than 4 hours (for example, 12 to 24 hours), 1/3 parts of bromoalkane is added for more than 1 to 4 hours, after the reaction, 1/3 parts of sodium hydride for more than 4 hours is added, 1/3 parts of bromoalkane is added for more than 1 to 4 hours, and finally the rest of sodium hydride for more than 4 hours is added until the reaction is finished.
The total reaction time of the step (2) is preferably 3-9 d.
Preferably, the acid in the step (3) is one or more of tetrafluoroboric acid, tetrabutylammonium fluoride and acetic acid.
In step (3), the amount of acid added needs to be excessive.
Preferably, the molar ratio of the c to the iodine simple substance and the triphenyl phosphine in the step (4) is 1:7 to 1:28 (1: 7 to 28). The reaction is carried out under anhydrous conditions.
Preferably, the alkali in steps (5) and (7) is one or more of sodium hydroxide, potassium hydroxide and sodium methoxide.
In the steps (5) and (7), the molar ratio of the reactant to the thiourea is 1: 3-1: 30.
Preferably, the molar ratio of the polyethylene glycol monomethyl ether to the phosphorus tribromide in the step (6) is 1: 2-1: 5.
Preferably, the oxidizing atmosphere in step (8) is realized by adding oxygen to the reaction solution, or adding an aqueous solution containing hydrogen peroxide or switching on air and heating.
The molar ratio of g to e is 1:7 to 1: 21. Within this range, it is helpful to prepare a hydrophilic segment that matches the hydrophobic segment, and further to improve the drug loading of the resulting carrier.
Preferably, in steps (1) - (8), the organic solvent is one or more selected from N, N-dimethylformamide, dichloromethane, methanol, ethanol and dimethyl sulfoxide.
Preferably, in steps (1) to (8), the anhydrous and oxygen-free reaction system is completed under an inert gas atmosphere such as nitrogen or argon.
The invention also discloses an application of the amphiphilic cyclodextrin polymer drug carrier, a hydrophobic drug and an organic solvent are mixed to obtain a dispersion, water is added into the dispersion, and self-assembly is carried out to obtain the hydrophobic drug-loaded sustained release preparation.
Preferably, the particle size of the amphiphilic cyclodextrin polymer drug carrier is preferably 40-100 nm. Under the particle size range, the effect of the obtained sustained-release preparation is favorably improved, and the side effect is less; for example, if an anti-tumor drug is loaded, the carrier with the particle size is favorable for enabling a drug-carrying system to be gathered at a tumor position through an EPR effect (namely the high permeability and long retention effect of solid tumors), so that the side effect is reduced, and the curative effect is improved.
Further preferably, the particle size of the amphiphilic cyclodextrin polymer drug carrier is preferably 40-50 nm. The carrier performance is more excellent in this preferred range.
The inventor finds that the self-polymerization method is helpful for further improving the drug loading rate and reducing the drug dissolution.
The inventor researches and discovers that the carrier and the self-polymerization method can obviously improve the drug loading capacity, avoid the burst effect and have excellent slow release performance.
The hydrophobic drug can be any water-insoluble compound drug.
According to the action part of the hydrophobic drug, a proper stimulus response chemical bond can be selected, so that the loaded drug is selectively released at the action part of the drug, and the action effect is improved.
Preferably, the hydrophobic drug is a hydrophobic anticancer drug. Preferably, the stimuli-responsive bond of the carrier for loading the hydrophobic anticancer drug is a redox stimuli-responsive bond; further preferred are disulfide bonds.
Preferably used for the carrier that hydrophobic tumor medicament loads, its 6-position hydroxy group connects polyglycol as the hydrophilic end, 2, 3-position hydroxy group connects the carbon chain as the hydrophobic end; the polyethylene glycol and the cyclodextrin are connected through a chemical bond with reduction stimulation responsiveness or DH stimulation responsiveness, and the bond can be directionally broken under a tumor tissue microenvironment. The amphiphilic cyclodextrin molecules can be self-assembled into a spherical drug-carrying system with a hydrophilic outer layer and a hydrophobic inner layer in a water phase, and hydrophobic anticancer drugs can be carried in a hydrophobic cavity. The amphiphilic cyclodextrin bouquet type polymer drug carrier has the advantages of large drug loading capacity, good biocompatibility, no toxic or side effect, directional release in a tumor tissue microenvironment, and good sustained and controlled release performance. The loaded medicine can be directionally released in a tumor tissue microenvironment, and has good sustained and controlled release performance, small side effect and high medicine effect.
According to the invention, through the self-polymerization of the carrier, a spherical structure with inner hydrophobicity and outer hydrophilicity is formed, so that the hydrophobic cavity of the cyclodextrin is enlarged, and the entrapment quantity of the hydrophobic drug is increased. In addition, a reduction-oxidation response or pH response type chemical bond such as a disulfide bond or an acylhydrazone bond is introduced on the amphiphilic cyclodextrin molecule, so that the amphiphilic cyclodextrin molecule can directionally release the anticancer drug in a tumor tissue microenvironment, and the drug content of a cancer tissue is increased, and the side effect of the carrier drug in a normal tissue is reduced.
Preferably, the hydrophobic anticancer drug is one or more of 5-fluorouracil, doxorubicin hydrochloride, hydroxycamptothecin, vincristine and paclitaxel.
In the application, the drug carrier is fully and uniformly mixed with the hydrophobic anticancer drug in the organic solvent, then the deionized water is slowly dropped in, the solvent is removed after self-assembly, and drying is carried out, thus forming the amphiphilic cyclodextrin spherical self-polymerization polymer drug-carrying system (also called as a sustained-release preparation).
In the invention, the obtained drug loading system can be subjected to drug loading test by adopting the conventional method. For example, amphipathic cyclodextrin drug-loaded systems are dissolved in methanol solution, sonicated, and the uv absorption is measured at a specific wavelength to yield the drug-loaded amount.
The invention also provides a controlled-release preparation of the hydrophobic drug, which is assembled by the application method and comprises spherical particles formed by self-assembly of a plurality of amphiphilic cyclodextrin polymer drug carriers, wherein the outer layer of the spherical particles is grafted with the hydrophilic chain segment; hydrophobic drugs are loaded in the inner cavity of the spherical particle, and the inner surface of the spherical particle is modified with a hydrophobic chain segment.
Taking the hydrophilic chain segment as PEG, the hydrophobic chain segment as dodecyl as an example, and the hydrophobic drug as an anti-tumor drug as an example, the structural formula is shown as formula 2:
the particle size of the amphiphilic cyclodextrin spherical self-polymerization system prepared by the invention is 40-100 nm, and the amphiphilic cyclodextrin spherical self-polymerization system can be further aggregated in tumor tissues through an EPR (ethylene propylene rubber) effect, so that a better treatment effect is achieved. The amphiphilic cyclodextrin drug-loaded system prepared by the method has the advantages of uniform particle size, controllable form, large drug-loaded amount, good stimulation responsiveness and high sustained and controlled release performance.
The drug loading of the sustained-release preparation can reach 40%.
Advantageous effects
The amphiphilic cyclodextrin bouquet type polymer drug carrier synthesized by the invention is prepared by taking cyclodextrin as a core and chemically modifying the cyclodextrin: the hydrophobic chain alkyl group is introduced to serve as a hydrophobic end of the amphiphilic polymer, so that the hydrophobic cavity of the cyclodextrin can be expanded to further increase the drug loading rate; the introduction of the stimulus response type chemical bond disulfide bond can directionally break in a reducing environment in a tumor tissue microenvironment, destroy a spherical self-assembly body formed by the amphiphilic cyclodextrin, and release the drug. Can reduce side effects in normal tissues and improve drug effects in tumor tissues. The introduction of the polyethylene glycol is used as a hydrophilic group of the amphiphilic cyclodextrin on one hand, is used for reducing the toxic and side effects of the material on the other hand, and can prolong the circulation time of the material in vivo. The amphiphilic cyclodextrin drug-loaded system prepared by the method has the particle size of 40-100 nm, and is beneficial to aggregation of the drug-loaded system at a tumor position through an EPR effect (namely a high-permeability and long-retention effect of solid tumors), so that the side effect is further reduced, and the curative effect is improved; the drug-loading system (also called as sustained release preparation) of the invention has drug-loading rate of 40%. Uniform grain diameter, controllable shape, large drug-loading rate, good stimulation responsiveness and high sustained and controlled release performance.
Drawings
FIG. 1 is a transmission electron microscope image of the amphipathic cyclodextrin spherical self-polymerized polymer drug delivery system prepared in example 1, and it can be seen that the spherical self-polymer of the present invention has a size of about 40nm to 50nm and a uniform size.
FIG. 2 is a nuclear magnetic hydrogen spectrum of the amphiphilic cyclodextrin polymer drug carrier prepared in example 1.The assignment of hydrogen of the polyethylene glycol and the cyclodextrin is between 3.38 and 3.83ppm, and the assignment of hydrogen of the dodecyl group is 1.26ppm (except-CH)3And α -H), 1.68ppm (. alpha. -H) and 0.88ppm (-CH3) in a ratio of 1358: 231: 28: 42, which indicates that both the primary (6-position) and secondary (2, 3-position) hydroxyl groups of the cyclodextrin are completely substituted.
FIG. 3 is the nuclear magnetic hydrogen spectrum of the amphiphilic cyclodextrin polymer drug carrier prepared in example 3. The assignment of hydrogen of the polyethylene glycol and the cyclodextrin is between 3.38 and 3.66ppm, and the assignment of hydrogen of the dodecyl group is 1.26ppm (except-CH)3And α -H), 1.68ppm (. alpha. -H) and 0.88ppm (-CH)3) The spectrum, at a ratio of 1358: 112: 21: 14, indicates that the primary hydroxyl groups of the cyclodextrin are completely substituted, but the secondary hydroxyl groups are only partially substituted.
FIG. 4 is a nuclear magnetic hydrogen spectrum of a cyclodextrin polymer drug carrier prepared in comparative example (comparative example 1). The hydrogen assignment of the polyethylene glycol and the cyclodextrin is between 3.24 and 3.75 ppm. The figures show that the primary hydroxyl groups of the cyclodextrin are substituted.
FIG. 5 is a cumulative release diagram of the controlled release of the drug in different environments (the drug is exemplified by doxorubicin) in the amphiphilic cyclodextrin spherical self-polymerization polymer drug delivery systems of examples 1-3 and comparative examples.
In FIG. 5, DTT is shown as the release profile after addition of the reducing agent Dithiothreitol (DTT), while DTT is shown as the release profile without addition of the reducing agent. The 8 icons in the figure represent the cumulative release profiles of DOX in examples 1-3 and comparative examples.
As can be seen from fig. 5, the drug delivery system prepared in example 1 has a significantly increased drug release rate in the presence of a reducing agent Dithiothreitol (DTT), and has little drug release in the absence of DTT. This fully demonstrates that the amphiphilic cyclodextrin spherical self-polymerization polymer drug-loading system of the invention has good controlled-release effect. The hydrophobic end in example 2 is ethyl, and the hydrophobic end is shortened relative to example 1, resulting in a relatively weak hydrophobic moiety, a small loading of drug, and a burst release. The water delivery chain in the example 3 only partially replaces the hydroxyl groups at the 2 and 3 positions of the cyclodextrin, and relatively speaking, the loading capacity of the drug is small. In the comparative case, cyclodextrin is hydrophobic relative to polyethylene glycol due to the absence of hydrophobic end, and would constitute a less stable self-polymerizing polymer, resulting in a more pronounced burst effect.
Detailed Description
The present invention will be described in detail with reference to specific examples, but the present invention is not limited thereto.
Example 1
(1) Protection of the 6-hydroxy group of cyclodextrins
In DMF, reacting 14.5g of tert-butyldimethylsilyl chloride, 9g of cyclodextrin and 5mL of triethylamine for 96 hours at 30 ℃ to protect 6-primary alcohol of cyclodextrin, then carrying out reduced pressure distillation to remove the solvent, carrying out gradient elution by using methanol and dichloromethane by a silica gel column chromatography to purify a product, and carrying out vacuum drying to obtain tert-butyldimethylsiloxy cyclodextrin a;
(2) alkylation of 2, 3-hydroxy groups of cyclodextrins
Taking 2g of a, firstly reacting with 0.5g of sodium hydride in anhydrous DMF for fully 24h, then adding 0.21mL of bromododecane, and reacting for 3d at 30 ℃; 0.5g of sodium hydride was added to the anhydrous DMF, and the mixture was reacted for 24 hours, followed by addition of 0.21mL of bromododecane and reaction at 30 ℃ for 2 days. Quenching with methanol, distilling under reduced pressure to remove solvent, purifying the product by gel chromatography, and vacuum drying to obtain 12-alkyl substituted tert-butyl dimethyl silica cyclodextrin b;
(3) deprotection of the 6-hydroxy group of cyclodextrins
Hydrolyzing 2g of b in an excessive tetrabutylammonium fluoride aqueous solution, removing tert-butyl dimethyl, neutralizing unreacted acid with ammonia water to neutrality, removing the solvent to obtain a crude product, and drying in vacuum to obtain a product c;
(4) iodination of 6-hydroxy cyclodextrin
Taking 2g of c to react with 6.47g of triphenylphosphine and 6.74g of iodine simple substance in anhydrous DMF at 70 ℃ for 24h, reducing the pressure and distilling to remove half of solvent, adjusting the pH value to 9 by using sodium methoxide, precipitating by using a large amount of methanol, washing by using methanol for three times to obtain a product, and drying to obtain d;
(5) thiolation of the 6-hydroxy group of cyclodextrin
Taking 2g of d, reacting with 0.57g of thiourea in DMF at 90 ℃ for 28h, distilling under reduced pressure to remove DMF, adding a proper amount of sodium hydroxide aqueous solution, refluxing for 1h, cooling the reaction solution, adjusting the pH to be less than 7 by using potassium hydrogen sulfate aqueous solution, extracting by using dichloromethane, washing by using saturated salt water, and removing the solvent to obtain a product e.
(6) Bromination of polyethylene glycol monomethyl ether
5g of polyethylene glycol monomethyl ether 2000 and 1mL of phosphorus tribromide are taken to react in anhydrous dichloromethane at the temperature of 20 ℃ for 24h, then the reaction solution is slowly dripped into water, dichloromethane is added for extraction, saturated saline water is used for washing to be neutral, drying is carried out, and a dichloromethane solvent is removed by a rotary evaporator to obtain bromo-polyethylene glycol monomethyl ether f.
(7) Thiolation of polyethylene glycol monomethyl ether
And (2) taking 5g of f and 5.7g of thiourea to react for 24 hours at 70 ℃ in anhydrous DMF, carrying out reduced pressure distillation to remove the organic solvent, adding a proper amount of aqueous solution of alkali, carrying out reflux reaction for 0.5-1.5 hours, cooling the reaction solution, adjusting the pH to be less than 7, extracting with dichloromethane, washing with saturated salt water, and carrying out reduced pressure distillation to remove the solvent to obtain g of thiolated polyethylene glycol monomethyl ether.
(8) Synthesis of amphiphilic cyclodextrin bouquet type polymer drug carrier
0.1g of e and 0.7g of g are taken to react well in DMSO solution in the presence of excess hydrogen peroxide to give the final amphiphilic cyclodextrin h.
(9) Assembly of amphiphilic cyclodextrin spherical self-polymerization polymer drug-loading system
And fully stirring and uniformly mixing 10mg of the mixture h and 10mg of adriamycin in DMSO, then dripping deionized water, removing the solvent after uniformly mixing, and drying to form the amphiphilic cyclodextrin drug-carrying system.
(10) Drug load measurement
The amphiphilic cyclodextrin drug-loaded system is dissolved in methanol solution, and the ultraviolet absorption is measured at 490nm by ultrasonic wave, so that the drug-loaded amount is 40% (mass/total mass of adriamycin).
(11) Sustained drug release study
The 10mg amphipathic cyclodextrin spherical self-polymerized polymer drug-loaded system is placed in 2mL PBS (or PBS containing 10M DTT), placed in a dialysis bag, then placed in a vial containing 8mL PBS buffer solution (or PBS containing 10M DTT), placed in a shaker, released at 37 ℃, and the outer layer of PBS is taken to measure ultraviolet at 490 nm. And obtaining the adriamycin cumulative release curve.
Example 2
Compared with the example 1, the main difference is that the hydrophobic chain segment is ethyl, and the specific operation is as follows:
(1) protection of the 6-hydroxy group of cyclodextrins
Reacting 9g of triphenylchloromethane, 5g of cyclodextrin and 5mL of triethylamine in DMF at 70 ℃ for 96 hours to protect primary alcohol at 6-position of cyclodextrin, then distilling under reduced pressure to remove the solvent, purifying the product by gradient elution of methanol and dichloromethane in silica gel column chromatography, and drying in vacuum to obtain triphenylmethyl ether-cyclodextrin a;
(2) alkylation of 2, 3-hydroxy groups of cyclodextrins
Taking 2g of a, firstly reacting with 0.5g of sodium hydride in anhydrous DMF for fully 24h, then adding 0.15mL of ethyl bromide, and reacting for 3d at 30 ℃; 0.5g of sodium hydride was added to anhydrous DMF and the mixture was reacted for 24 hours, followed by addition of 0.15mL of ethyl bromide and reaction at 30 ℃ for 2 days. Quenching with methanol, distilling under reduced pressure to remove solvent, purifying the product by gel chromatography, and vacuum drying to obtain methoxy-triphenylmethoxy-cyclodextrin b;
(3) steps (9) example 1 was repeated
(10) Drug load measurement
The amphiphilic cyclodextrin drug-loaded system is dissolved in methanol solution, and the ultraviolet absorption of the amphiphilic cyclodextrin drug-loaded system is measured at specific wavelength by ultrasonic wave, so that the drug-loaded rate is 25 percent.
(11) Sustained drug release study
Placing 5-10 mg of amphiphilic cyclodextrin spherical self-polymerization polymer drug delivery system into 2mL of PBS (or PBS containing 10 MDTT), placing the system into a dialysis bag, then placing the system into a vial containing 8mL of PBS buffer solution (or PBS containing 10M DTT), placing the system into a shaker, releasing the system at 37 ℃, and taking the outer layer of PBS to measure ultraviolet at a specific wavelength. Obtaining the cumulative release curve of the anticancer drug.
Example 3
Compared with the example 1, the main difference is that the hydrophobic chain segment is modified at the 2, 3-position of the part, and the specific operation is as follows:
(1) protection of the 6-hydroxy group of cyclodextrins
In DMF, reacting 14.5g of tert-butyldimethylsilyl chloride, 9g of cyclodextrin and 5mL of triethylamine for 96 hours at 70 ℃ to protect 6-primary alcohol of cyclodextrin, then carrying out reduced pressure distillation to remove the solvent, carrying out gradient elution by using methanol and dichloromethane by a silica gel column chromatography to purify a product, and carrying out vacuum drying to obtain tert-butyldimethylsiloxy cyclodextrin a;
(2) alkylation of 2, 3-hydroxy groups of cyclodextrins
Taking 2g of a, firstly, fully reacting with 0.5g of sodium hydride in anhydrous DMF, then adding 0.21mL of bromododecane, reacting at 30 ℃ for 24h, quenching with methanol, distilling under reduced pressure to remove the solvent, purifying the product by a gel chromatography separation method, and drying in vacuum to obtain 12-alkyl substituted tert-butyl dimethyl silica cyclodextrin b;
(3) deprotection of the 6-hydroxy group of cyclodextrins
Hydrolyzing 2g of b in an excessive tetrabutylammonium fluoride aqueous solution, removing tert-butyl dimethyl, neutralizing unreacted acid with ammonia water to neutrality, removing the solvent to obtain a crude product, and drying in vacuum to obtain a product c;
(4) iodination of 6-hydroxy cyclodextrin
Taking 2g of c to react with 3.24g of triphenylphosphine and 3.37g of iodine simple substance in anhydrous DMF at 70 ℃ for 24h, reducing the pressure and distilling to remove half of solvent, adjusting the pH value to 9 by using sodium methoxide, precipitating by using a large amount of methanol, washing by using methanol for three times to obtain a product, and drying to obtain d;
(5) thiolation of the 6-hydroxy group of cyclodextrin
Taking 2g of d, reacting with 0.29g of thiourea in DMF at 90 ℃ for 28h, distilling under reduced pressure to remove DMF, adding a proper amount of sodium hydroxide aqueous solution, refluxing for 1h, cooling the reaction solution, adjusting the pH to be less than 7 by using potassium hydrogen sulfate aqueous solution, extracting by using dichloromethane, washing by using saturated salt water, and removing the solvent to obtain a product e.
(6) Bromination of polyethylene glycol monomethyl ether
5g of polyethylene glycol monomethyl ether 2000 and 1mL of phosphorus tribromide are taken to react in anhydrous dichloromethane at the temperature of 20 ℃ for 24h, then the reaction solution is slowly dripped into water, dichloromethane is added for extraction, saturated saline water is used for washing to be neutral, drying is carried out, and a dichloromethane solvent is removed by a rotary evaporator to obtain bromo-polyethylene glycol monomethyl ether f.
(7) Thiolation of polyethylene glycol monomethyl ether
And (2) taking 5g of f and 5.7g of thiourea to react for 24 hours at 70 ℃ in anhydrous DMF, carrying out reduced pressure distillation to remove the organic solvent, adding a proper amount of aqueous solution of alkali, carrying out reflux reaction for 0.5-1.5 hours, cooling the reaction solution, adjusting the pH to be less than 7, extracting with dichloromethane, washing with saturated salt water, and carrying out reduced pressure distillation to remove the solvent to obtain g of thiolated polyethylene glycol monomethyl ether.
(8) Synthesis of amphiphilic cyclodextrin bouquet type polymer drug carrier
0.1g of e and 0.7g of g are taken to react well in DMSO solution in the presence of excess hydrogen peroxide to give the final amphiphilic cyclodextrin h.
(9) Assembly of amphiphilic cyclodextrin spherical self-polymerization polymer drug-loading system
And fully stirring and uniformly mixing 10mg of the mixture h and 10mg of adriamycin in DMSO, slowly dripping deionized water, removing the solvent after uniformly mixing, and drying to form the amphiphilic cyclodextrin drug-carrying system.
(10) Drug load measurement
The amphiphilic cyclodextrin drug-loaded system is dissolved in methanol solution, and the ultraviolet absorption is measured at 490nm by ultrasonic wave, so that the drug-loaded amount is 29% (mass/total mass of adriamycin).
(11) Sustained drug release study
The 10mg amphipathic cyclodextrin spherical self-polymerized polymer drug-loaded system is placed in 2mL PBS (or PBS containing 10M DTT), placed in a dialysis bag, then placed in a vial containing 8mL PBS buffer solution (or PBS containing 10M DTT), placed in a shaker, released at 37 ℃, and the outer layer of PBS is taken to measure ultraviolet at 490 nm. And obtaining the adriamycin cumulative release curve.
Comparative example (comparative example 1)
Compared with the examples, the difference is that the carrier which is relatively close to the carrier of the prior art and is only modified at the 6-position is prepared, and the specific operation is as follows:
(1) iodination of primary hydroxyl groups of cyclodextrins
Taking 2g of cyclodextrin to react with 6.47g of triphenylphosphine and 6.74g of iodine simple substance in anhydrous DMF at 70 ℃ for 24h, reducing the pressure and distilling to remove half of solvent, adjusting the pH value to 9 by using sodium methoxide, precipitating by using a large amount of methanol, washing by using methanol for three times to obtain a product, and drying to obtain a;
(2) thiolation of primary cyclodextrin hydroxyls
Taking 2g of a, reacting with 0.57g of thiourea in DMF at 90 ℃ for 28h, distilling under reduced pressure to remove DMF, adding a proper amount of sodium hydroxide aqueous solution, refluxing for 1h, cooling the reaction solution, adjusting the pH to be less than 7 by using potassium hydrogen sulfate aqueous solution, extracting by using dichloromethane, washing by using saturated salt water, and removing the solvent to obtain a product c.
(3) Bromination of polyethylene glycol monomethyl ether
5g of polyethylene glycol monomethyl ether 2000 and 1mL of phosphorus tribromide are taken to react in anhydrous dichloromethane at the temperature of 20 ℃ for 24h, then the reaction solution is slowly dripped into water, dichloromethane is added for extraction, saturated saline water is used for washing to be neutral, drying is carried out, and a dichloromethane solvent is removed by a rotary evaporator to obtain bromo-polyethylene glycol monomethyl ether f.
(4) Thiolation of polyethylene glycol monomethyl ether
And (2) taking 5g of f and 5.7g of thiourea to react for 24 hours at 70 ℃ in anhydrous DMF, carrying out reduced pressure distillation to remove the organic solvent, adding a proper amount of aqueous solution of alkali, carrying out reflux reaction for 0.5-1.5 hours, cooling the reaction solution, adjusting the pH to be less than 7, extracting with dichloromethane, washing with saturated salt water, and carrying out reduced pressure distillation to remove the solvent to obtain g of thiolated polyethylene glycol monomethyl ether.
(5) Synthesis of cyclodextrin polymer drug carriers
0.05g of e and 0.7g of g are taken to react well in DMSO solution in the presence of excess hydrogen peroxide to give the final cyclodextrin polymer h.
(6) Assembly of cyclodextrin polymer drug delivery system
And (3) fully and uniformly stirring 10mg of the mixture h and 10mg of adriamycin in DMSO, slowly dripping deionized water, uniformly mixing, removing the solvent, and drying to form a cyclodextrin drug-loaded system.
(7) Drug load measurement
The cyclodextrin drug-loaded system was dissolved in methanol solution, and the ultraviolet absorption was measured at 490nm with ultrasound to obtain a drug-loaded amount of 21% (mass/total mass of doxorubicin).
(8) Sustained drug release study
The 10mg amphipathic cyclodextrin spherical self-polymerized polymer drug-loaded system is placed in 2mL PBS (or PBS containing 10M DTT), placed in a dialysis bag, then placed in a vial containing 8mL PBS buffer solution (or PBS containing 10M DTT), placed in a shaker, released at 37 ℃, and the outer layer of PBS is taken to measure ultraviolet at 490 nm. And obtaining the adriamycin release curve.
Claims (18)
1. An amphiphilic cyclodextrin polymer drug carrier, characterized in that: taking cyclodextrin as a core, wherein the 6-position of each sugar ring of the cyclodextrin core is modified and grafted with a hydrophilic chain segment; 2-position and 3-position of each sugar ring are modified with hydrophobic chain segments;
the hydrophilic chain segment is a polyethylene glycol hydrophilic chain segment; the hydrophilic chain segment is connected with the 6-position of the cyclodextrin through a stimulus response type chemical bond; the stimulus-responsive chemical bond is a redox stimulus-responsive chemical bond; the redox stimulus responsive chemical bond is a disulfide bond;
the hydrophobic chain segment is dodecyl.
2. The amphiphilic cyclodextrin polymer drug carrier of claim 1, wherein: the molecular weight of the polyethylene glycol hydrophilic chain segment is 200-8000.
3. A method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 1 or 2, wherein: the method comprises the following steps:
step (a): protection of the 6-hydroxy group of cyclodextrins
Protecting 6-primary alcohol of cyclodextrin to obtain 6-hydroxyl protected cyclodextrin a;
step (b): hydrophobic modification of hydroxyl groups at positions 2 and 3 of cyclodextrin;
reacting a with a source material of a hydrophobic chain segment under alkali to obtain a 2-position and 3-position hydroxyl hydrophobicity modified product b;
step (c): deprotection of the hydroxyl group at position 6 of product b
Hydrolyzing the b under an acidic condition, and removing the primary hydroxyl protecting group at the 6 th position to obtain a product c;
(d) and connecting the product c with a stimulus-responsive chemical bond to prepare the carrier, wherein the stimulus-responsive chemical bond is a disulfide bond.
4. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 3, wherein: the method comprises the following steps:
(1) protection of the 6-hydroxy group of cyclodextrins
In an organic solvent, reacting a primary hydroxyl protecting group, cyclodextrin and triethylamine for 72-120 h at 20-80 ℃ to protect 6-primary alcohol of cyclodextrin, and then removing the solvent to obtain 6-hydroxyl protected cyclodextrin a;
(2) alkylation of 2, 3-hydroxy groups of cyclodextrins
Fully reacting a with sodium hydride in an organic solvent, adding bromoalkane, reacting for 2-5 days at 0-50 ℃, quenching, removing the solvent, purifying the product by a gel chromatography separation method, and drying to obtain a product b; the chemical formula of the bromoalkane is R1-Br, said R1Is dodecyl; a, bromoalkane and sodium hydride in a molar ratio of 1:7:7 to 1:60: 60;
(3) deprotection of the 6-hydroxy group of cyclodextrins
Hydrolyzing the b under an acidic condition, removing a primary hydroxyl protecting group, neutralizing unreacted acid with ammonia water, removing the solvent to obtain a crude product, and drying in vacuum to obtain a product c;
(4) iodination of 6-hydroxy cyclodextrin
c, reacting with triphenyl phosphorus and iodine simple substances in an anhydrous organic solvent at 60-90 ℃ for 18-36 h, reducing half of the solvent, adjusting the pH value to 9-10 by using sodium methoxide, precipitating by using a large amount of methanol, washing by using methanol for three times to obtain a product, and drying to obtain d;
(5) thiolation of the 6-hydroxy group of cyclodextrin
d, reacting the thiourea in an organic solvent at 60-100 ℃ for 16-28 h, removing the organic solvent, adding a proper amount of aqueous solution of alkali, performing reflux reaction for 0.5-1.5 h, cooling the reaction solution, adjusting the pH to be less than 7, extracting with dichloromethane, washing with saturated salt water, and removing the solvent to obtain a product e;
(6) bromination of polyethylene glycol monomethyl ether
Reacting polyethylene glycol monomethyl ether with phosphorus tribromide in an anhydrous organic solvent at 0-30 ℃ for 18-28 h, slowly dropwise adding the reaction solution into water, adding dichloromethane for extraction, washing with saturated saline water to be neutral, drying, and removing the dichloromethane solvent by using a rotary evaporator to obtain bromo-polyethylene glycol monomethyl ether f;
(7) thiolation of polyethylene glycol monomethyl ether
f, reacting the mixture with thiourea in an anhydrous organic solvent at 60-100 ℃ for 16-28 h, removing the organic solvent, adding a proper amount of alkali aqueous solution, carrying out reflux reaction for 0.5-1.5 h, cooling the reaction solution, adjusting the pH to be less than 7, extracting with dichloromethane, washing with saturated salt water, removing the solvent to obtain thiolated polyethylene glycol monomethyl ether g;
(8) synthesis of amphiphilic cyclodextrin bouquet type polymer drug carrier
g. e fully reacting in an oxidizing environment to obtain the final amphiphilic cyclodextrin h.
5. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: in the step (1), the molar ratio of cyclodextrin to primary hydroxyl protecting group is 1: 3-1: 14; the primary hydroxyl protecting group is one or more of triphenylchloromethane, tert-butyldimethylchlorosilane and p-toluenesulfonyl chloride.
6. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: in the step (2), bromoalkane and NaH are added in batches, the batch adding method comprises the steps of dividing sodium hydride and bromoalkane into 2-3 parts respectively, adding one part of sodium hydride firstly, reacting for more than 4 hours, adding one part of bromoalkane again, and repeating the step after reacting for 1-4 days until the reaction is finished.
7. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: the acid in the step (3) is one or more of tetrafluoroboric acid, tetrabutylammonium fluoride and acetic acid; the amount of acid added needs to be in excess.
8. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: in the step (4), the molar ratio of the c to the iodine simple substance and the triphenyl phosphine is 1:7: 7-1: 28: 28.
9. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: the alkali in the steps (5) and (7) is one or more of sodium hydroxide, potassium hydroxide and sodium methoxide; the molar ratio of the reactants to the thiourea is 1: 3-1: 30.
10. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: in the step (6), the molar ratio of the polyethylene glycol monomethyl ether to the phosphorus tribromide is 1: 2-1: 5.
11. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: the oxidizing environment in the step (8) is realized by adding oxygen into the reaction solution, or adding an aqueous solution containing hydrogen peroxide or switching on air and heating; the molar ratio of g to e is 1:7 to 1: 21.
12. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: in the steps (1) - (8), the organic solvent is one or more selected from N, N-dimethylformamide, dichloromethane, methanol, ethanol and dimethyl sulfoxide.
13. The method of preparing the amphiphilic cyclodextrin polymer drug carrier of claim 4, wherein: in the steps (1) to (8), the anhydrous anaerobic reaction system is in a nitrogen or argon atmosphere.
14. The use of an amphiphilic cyclodextrin polymer drug carrier of any of claims 1-2 in the preparation of a sustained release formulation, wherein: mixing the amphiphilic cyclodextrin polymer drug carrier, the hydrophobic drug and the organic solvent to obtain dispersion, adding water into the dispersion, and carrying out self-assembly to obtain the hydrophobic drug-loaded sustained-release preparation.
15. The use of claim 14, wherein: the hydrophobic drug is a hydrophobic anticancer drug.
16. The use of claim 15, wherein: the hydrophobic drug is one or more of 5-fluorouracil, doxorubicin hydrochloride, hydroxycamptothecin, vincristine and paclitaxel.
17. The use of claim 14, wherein: the particle size of the amphiphilic cyclodextrin polymer drug carrier is 40-100 nm.
18. A controlled release preparation of a hydrophobic drug, which is assembled by the application method of any one of claims 14 to 17, and comprises spherical particles formed by self-assembly of a plurality of amphiphilic cyclodextrin polymer drug carriers, wherein the outer layer of the spherical particles is grafted with the hydrophilic chain segment; hydrophobic drugs are loaded in the inner cavity of the spherical particles, and the inner surfaces of the spherical particles are modified with hydrophobic chain segments.
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