CN116178589A - Amphiphilic lipoyl cationic chitosan derivative and preparation and application thereof - Google Patents
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Abstract
The invention relates to the technical field of nano biomedicine, in particular to an amphiphilic lipoyl cationic chitosan derivative, a preparation method and application thereof. The structural formula of the amphiphilic lipoic acid acyl cationic chitosan derivative is shown as a formula (1), wherein R is pyridinium containing lipoic acid amide, the average polymerization degree n is 20-60, the steps for preparing the amphiphilic compound are simple and easy, the nano micelle prepared by the amphiphilic lipoic acid acyl cationic chitosan derivative has good biocompatibility, has higher entrapment capacity and redox responsive release behavior for an antitumor drug, and is a carrier with good application value.
Formula (1).
Description
Technical Field
The invention belongs to the technical field of nano biomedicine, and particularly relates to an amphiphilic lipoyl cationic chitosan derivative, a preparation method thereof and application thereof in nano carriers.
Background
Chitosan (Chitosan) is an alkaline polysaccharide obtained by deacetylation of chitin, and is a renewable polysaccharide with no toxic or side effect and good biocompatibility and degradability, and has a plurality of unique physiological and pharmacological functional properties, so that the Chitosan has great application potential in the field of self-assembled nano drug delivery systems. The chitosan itself has certain hydrophilicity, but lacks a lipophilic structure and cannot form a nano micelle with a core-shell structure, the chitosan itself contains amino and hydroxyl groups which can be used as chemical modification sites, and hydrophobic groups can be introduced into the chitosan by a high-efficiency chemical modification means to obtain amphiphilic compounds, so that the amphiphilic compounds are prepared into nano carriers. In addition, the stability of the nano-carrier is closely related to the charge of the micelle, the 2-amino group in the chitosan molecule is easy to be protonated in an acidic solution, and the chitosan is electropositive under an acidic condition, but cannot show electropositive under neutral and alkaline conditions; meanwhile, the amino group at the 2-position of chitosan is relatively active, and is easy to chemically modify, and the electropositivity of the modified chitosan is greatly reduced, so that stable positive charges are required to be introduced to improve the stability of the chitosan nano-micelle. Meanwhile, considering the redox responsiveness of disulfide bonds to the tumor microenvironment, the invention utilizes the lipophilic structure of lipoic acid and the redox responsiveness of disulfide bonds to connect chitosan and lipoic acid through pyridine cations to form a lipoic acid cation amphiphilic compound, and further prepares the redox responsive nano micelle.
Disclosure of Invention
The invention aims to provide an amphiphilic lipoyl cationic chitosan derivative which can be used as a carrier of a hydrophobic chemotherapeutic drug, and preparation and application thereof.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
an amphiphilic lipoyl cationic chitosan derivative, the structural formula of which is shown in the formula (1),
formula (1), wherein->
Wherein the value range of the polymerization degree n is 20-60.
The preparation method of the amphiphilic lipoyl cationic chitosan derivative comprises the following steps: firstly, the chitosan is subjected to nicotinoylation, and then nucleophilic reaction with bromoethylamine hydrobromide to obtain aminoethyl cationic chitosan, and the obtained aminoethyl cationic chitosan is used in the following steps ofN,NThe' -carbonyl diimidazole reacts with alpha-lipoic acid under the catalysis of the carbonyl diimidazole to obtain the amphiphilic lipoic acyl cationic chitosan derivative shown in the formula (1);
the saidN,NThe molar quantity of the' -carbonyl diimidazole is 3.0-6.0 times of that of the chitosan; the molar quantity of the nicotinic acid is 3.0-6.0 times of that of the chitosan; the molar quantity of the bromoethylamine hydrobromide is 2.0-5.0 times of that of the nicotinoylated chitosan;N,Nthe molar quantity of the' -carbonyl diimidazole is 1.0-4.0 times of the molar quantity of the alpha-lipoic acid; the molar quantity of the alpha-lipoic acid is 1.0 to 4.0 times of that of the aminoethyl cation chitosan.
Will beN,NAdding' -carbonyl diimidazole into dimethyl sulfoxide solution containing nicotinic acid, then reacting with chitosan at 50-70 ℃ for 12-24 h, precipitating the reaction mixture with excessive acetone, washing, cooling and drying to obtain nicotinoylated chitosan; reacting the obtained nicotinamide acylated chitosan with bromoethylamine hydrobromide at 50-70 ℃ for 12-36 h, precipitating the obtained solid with ethanol, washing, and freeze-drying to obtain the aminoethyl cationic chitosan for later use.
Will beN,N'-Adding carbonyl diimidazole into dimethyl sulfoxide solution containing alpha-lipoic acid, and reacting at 50-70 ℃ in nitrogen atmosphere for 12-36 h, wherein the obtained solution is prepared for standby; dissolving the obtained aminoethyl cationic chitosan in dimethyl sulfoxide, continuously reacting with the obtained solution at 50-70 ℃ for 12-36 h, then precipitating and washing with acetone, and freeze-drying to obtain the amphiphilic lipoyl cationic chitosan derivative shown in the formula (1).
The application of the amphiphilic lipoyl cationic chitosan derivative is that the nano micelle prepared by the amphiphilic lipoyl cationic chitosan derivative shown in the formula (1) is applied to a drug carrier.
The invention has the advantages that:
(1) The preparation method of the amphiphilic lipoyl cationic chitosan derivative has the advantages of simple and efficient process, easy acquisition of required equipment and raw materials, low cost, safety and innocuity, good biocompatibility and suitability for being applied to the field of biological medicine.
(2) The redox responsive nano micelle prepared by the amphiphilic lipoyl cationic chitosan derivative has positive surface net charge, so that the redox responsive nano micelle can penetrate through a cell membrane to enter cells in a cell endocytosis mode, and the entrapped chemotherapeutic medicine enters the cells to exert the efficacy, and has congenital advantages in the aspect of medicine transmission.
(3) The redox responsive nano micelle prepared by the amphiphilic lipoyl cationic chitosan derivative has good entrapment capacity and redox behavior, can passively target a hydrophobic chemotherapeutic drug to tumor cells, disintegrate in a high-concentration Glutathione (GSH) environment in the tumor cells, can realize the rapid release of the chemotherapeutic drug in the tumor cells, improves the curative effect of the chemotherapeutic drug, and is a carrier with good application value.
Drawings
FIG. 1 is an infrared spectrum of chitosan.
FIG. 2 is an infrared spectrum of a nicoti-acylated chitosan provided by an embodiment of the present invention, 1726 cm -1 Absorption peak as ester bond, 1650 cm -1 Absorption peaks for amide bonds, 829, 743, 703 and 630 cm -1 Is the absorption peak of the pyridine group.
FIG. 3 is an infrared spectrum of an aminoethyl cationic chitosan according to an embodiment of the present invention, which is shown in FIG. 2, 1503 cm -1 The successful synthesis of aminoethyl cationic chitosan was demonstrated as an absorption peak for pyridine cations.
FIG. 4 is an infrared spectrum of an amphiphilic lipoyl cationic chitosan derivative according to an embodiment of the present invention, which is the same as that of FIG. 3Compared with ethyl cationic chitosan, 2930 and 2930 cm -1 And 2863 cm -1 As the absorption peak of the lipoic acid alkyl chain, the analysis data above prove the successful synthesis of the amphiphilic lipoic acid acyl cationic chitosan derivative.
Fig. 5 is a graph of the experimental result of preparing nano-micelle by using amphiphilic lipoyl cationic chitosan derivative, wherein the physical graph in the graph represents the measurement result of the hemolysis rate of nano-micelle prepared by using amphiphilic lipoyl cationic chitosan derivative under different concentrations by using distilled water as a positive control and physiological saline as a negative control.
Fig. 6 is a graph showing the results of in vitro release experiments of drug-loaded micelles prepared from amphiphilic lipoyl cationic chitosan derivatives, wherein p <0.01 shows that the cumulative release amounts of doxorubicin have obvious statistical differences under two different conditions of ph=5.5 and ph=5.5+gsh.
Detailed Description
The invention is further illustrated by the following figures and examples.
According to the invention, pyridine cations and lipoic acid are introduced to synthesize the redox-response amphiphilic compound, and the amphiphilic compound simultaneously contains hydrophilic chains and hydrophobic chains, so that the redox-response nano micelle can be self-assembled; the amphiphilic compound prepared by the invention has simple and easy steps, the nano micelle prepared by the amphiphilic lipoyl cationic chitosan derivative has good biocompatibility, has higher entrapment capacity and redox responsive release behavior for anti-tumor drugs, and is a carrier with good application value.
Specifically: first usingN,N'Activating carboxyl on nicotinic acid molecule with carbonyl diimidazole, then reacting with chitosan to obtain nicotinamide acylated chitosan, and then reacting with bromoethylamine hydrobromic acid to obtain aminoethyl cation chitosan, wherein the obtained aminoethyl cation chitosan is prepared by nucleophilic reactionN,NThe' -carbonyl diimidazole reacts with the lipoic acid serving as a redox response group under the catalysis of the carbonyl diimidazole to obtain the amphiphilic lipoic acid cation chitosan derivative.
The nano micelle prepared from the amphiphilic lipoyl cationic chitosan derivative is a drug carrier with good application value, and the prepared drug-loaded nano micelle can passively target tumor cells and rapidly release drugs in the tumor cells, so that high-efficiency drug release is realized, and the clinical curative effect of the chemotherapeutic drugs is improved.
The synthesis route of the amphiphilic lipoyl cationic chitosan derivative is as follows:
wherein R is 1 Is nicotinyl, R 2 Is bromoethylamine pyridine cation, R is pyridine salt containing sulfur octanoyl amide, and the average polymerization degree n is 20-60.
Example 1
The amphiphilic lipoyl cationic chitosan derivative of the target compound is synthesized according to the synthetic route.
1) Preparation of nicotinamide chitosan: weigh 7.38 g (60 mmol) nicotinic acid into 10 mL dimethyl sulfoxide solution and then add 9.72 g (60 mmol)N,NAfter stirring for 30 min, the' -carbonyl diimidazole is reacted at 60 ℃ under the protection of nitrogen for 12 h for later use. 3.22. 3.22 g (20 mmol) of chitosan (see FIG. 1) was weighed and dissolved in 30 mL dimethyl sulfoxide, then the solution prepared in the previous step was dropped, and the reaction was stirred under nitrogen protection at 60℃for 12 h. And then precipitating with excessive acetone, washing, and freeze-drying to obtain nicotinoylated chitosan 2.66 g (see figure 2) for later use.
2) Preparation of aminoethyl cationic Chitosan 1.596 g (6 mmol) of nicotinamide acylated chitosan was weighed into a solution of 20 mL dimethyl sulfoxide, then 2.46 g (12 mmol) of bromoethylamine hydrobromide was added to the solution in N 2 The reaction was stirred at 60℃for 12 h under protection. After the reaction, the reaction solution was dropped into ethanol and repeatedly washed for 3 times, and the solid was obtained by suction filtration, and the solid was dried in vacuo to obtain aminoethyl cationic chitosan 2.02. 2.02 g (see fig. 3).
3) Preparation of amphiphilic lipoyl cationic chitosan derivatives: 0.83 g (4 mmol) of alpha-sulfur Xin Suanrong is weighed into 2 mL dimethyl sulfoxide solution, and 0.65 g (4) mmol)N,N' -carbonyl diimidazole, activating carboxyl 12 h at 60 ℃ to obtain solution A; 1.6 g (4 mmol) of aminoethyl nicotinoyl chitosan is weighed and dissolved in 5 mL dimethyl sulfoxide solution to obtain solution B; then add solution B to solution A, at N 2 Under the protection of the method, stirring at 60 ℃ to continue the reaction for 12 h, after the reaction is finished, dripping the reaction solution into acetone to wash and precipitate for 3 times, filtering to obtain a solid, and drying the solid in vacuum to obtain the amphiphilic lipoyl cationic chitosan derivative 1.278 g shown in the formula (1) (see figure 4), wherein the average polymerization degree n is in the range of 20-60.
Example 2
1) Preparation of nicotinamide chitosan: 9.84 g (80 mmol) of nicotinic acid was weighed into 15 mL of dimethyl sulfoxide solution, followed by 12.96 g (80 mmol)N,NAfter stirring for 40 min, the' -carbonyl diimidazole is reacted at 70 ℃ under the protection of nitrogen for 24 h for later use. 3.22. 3.22 g (20 mmol) of chitosan (see FIG. 1) was weighed and dissolved in 40 mL dimethyl sulfoxide, then the solution prepared in the previous step was dropped, and the reaction was stirred under nitrogen protection at 70℃for 24 h. And then precipitating with excessive acetone, washing, and freeze-drying to obtain nicotinoylated chitosan 2.87. 2.87 g (see figure 2) for later use.
2) Preparation of aminoethyl cationic Chitosan 1.596 g (6 mmol) of nicotinamide acylated chitosan was weighed into a solution of 20 mL dimethyl sulfoxide, then 3.072 g (15 mmol) of bromoethylamine hydrobromide was added to the solution in N 2 Reaction 18 h was stirred at 65℃under protection. After the reaction, the reaction solution was dropped into ethanol to be repeatedly washed for 3 times, and the solid was obtained by suction filtration and dried in vacuo to obtain aminoethyl cationic chitosan 2.56. 2.56 g (see fig. 3).
3) Preparation of amphiphilic lipoyl cationic chitosan derivatives: 1.24 g (6 mmol) of alpha-sulfur Xin Suanrong was weighed into 2 mL dimethyl sulfoxide solution and 0.97 g (6 mmol) was slowly addedN,N' -carbonyl diimidazole, activating carboxyl 18 h at 65 ℃ to obtain solution A; 1.6 g (4 mmol) of aminoethyl nicotinoyl chitosan is weighed and dissolved in 5 mL dimethyl sulfoxide solution to obtain solution B; then add solution B to solution A, at N 2 Under the protection, stirring at 65 ℃ to continue the reaction for 18 h, and dripping the reaction liquid into acetone after the reaction is finishedThe precipitate is washed 3 times, the solid is obtained by suction filtration, and the solid is dried in vacuum to obtain 1.46g (see figure 4) of amphiphilic lipoyl cationic chitosan derivative shown in the formula (1), wherein the average polymerization degree n is in the range of 20-60.
Example 3
1) Preparation of nicotinamide chitosan: 12.3. 12.3 g (100 mmol) of nicotinic acid was weighed into 30 mL dimethyl sulfoxide solution, followed by 16.2. 16.2 g (100 mmol)N,NThe' -carbonyl diimidazole is stirred for 60 min and then reacted at 80 ℃ for 36 h under the protection of nitrogen for later use. 3.22. 3.22 g (20 mmol) of chitosan (see FIG. 1) was weighed and dissolved in 60 mL dimethyl sulfoxide, then the solution prepared in the previous step was dropped, and the reaction was stirred under nitrogen protection at 80℃for 36 h. And then precipitating with acetone, washing, and freeze-drying to obtain the nicotinoylated chitosan 3.66 g (see figure 2) for later use.
2) Preparation of aminoethyl cationic Chitosan 1.596 g (6 mmol) of nicotinamide acylated chitosan was weighed into a solution of 20 mL dimethyl sulfoxide, then 3.686 g (18 mmol) of bromoethylamine hydrobromide was added to N 2 Reaction 24 h was stirred at 80℃under protection. After the reaction, the reaction solution was dropped into ethanol to be repeatedly washed for 3 times, and the solid was obtained by suction filtration, and the solid was dried in vacuo to obtain aminoethyl cationic chitosan 3.01. 3.01 g (see fig. 3).
3) Preparation of amphiphilic lipoyl cationic chitosan derivatives: 1.65 g (8 mmol) of alpha-sulfur Xin Suanrong was weighed into 2 mL dimethyl sulfoxide solution and 1.30 g (8 mmol) was slowly addedN,N' -carbonyl diimidazole, activating carboxyl 24 h at 80 ℃ to obtain solution A; 1.6 g (4 mmol) of aminoethyl nicotinoyl chitosan is weighed and dissolved in 5 mL dimethyl sulfoxide solution to obtain solution B; then add solution B to solution A, at N 2 Under the protection of the method, stirring at 80 ℃ to continue the reaction for 24 h, after the reaction is finished, dripping the reaction solution into acetone to wash and precipitate for 3 times, filtering to obtain a solid, and drying the solid in vacuum to obtain the amphiphilic lipoyl cationic chitosan derivative shown in the formula (1) 1.83 g (see figure 4), wherein the average polymerization degree n is in the range of 20-60.
Application example 1
Preparation of nano micelle by amphiphilic lipoyl cationic chitosan derivative
(1) The nano micelle is prepared by a dialysis/ultrasonic method. Dissolving the product of the example 1 in dimethyl sulfoxide with the concentration of 2 mg/mL, transferring the solution into a dialysis bag (Mwco=100), dialyzing in water for 24 h at room temperature, and placing the dialyzate in a probe ultrasonic instrument (135W) for ultrasonic treatment (30 min) after the dialyzate is completely dialyzed, thus obtaining the redox stimulus response type cation nano micelle.
(2) Characterization of the preparation of cationic nanomicelle from amphiphilic lipoyl cationic chitosan derivative
The resulting redox-responsive micelles were characterized using Dynamic Light Scattering (DLS). Placing 1 mL of the redox stimulus-responsive cationic nano micelle prepared in the step (1) in a cuvette, and measuring the particle size, potential and PDI value in a Litesizer 500 nanometer particle size measuring instrument;
experimental results: the characterization of the preparation of the cationic nano-micelle by the amphiphilic lipoic acid cationic chitosan derivative synthesized in the embodiment 1 is shown in the table 1, the particle size of the nano-micelle prepared by the amphiphilic lipoic acid cationic chitosan derivative is 289.01 +/-4.94 nm, the particle size is less than 500 nm, the clearance of a blood system can be avoided, and the nano-micelle can be better phagocytosed by cells in a focus area to further play a role in entering the cells; PDI is 25.54+/-1.52 and less than 30, which indicates that the prepared nano micelle is uniformly distributed; the potentials were all positive at 42.14.+ -. 0.60. 0.60 mV, which is closely related to the presence of pyridine cations, further demonstrating the successful preparation of nicotinamide chitosan oligosaccharides.
TABLE 1 particle size, PDI and potential test results of cationic nanomicelles prepared in EXAMPLE 1
(3) Safety evaluation was performed on the cationic nanomicelle prepared according to step (1) using the amphiphilic lipoyl cationic chitosan derivative synthesized in example 1.
The method comprises the following steps: firstly, taking 5 mL of healthy rabbit blood, centrifuging at 2500 rpm for 10 min, discarding supernatant plasma, adding physiological saline, mixing uniformly, and centrifuging again under the same condition. This procedure was repeated three times until the supernatant was colorless, and a suspension of 2% strength erythrocytes was obtained by adding physiological saline. Preparing nano micelle solutions (2, 1.5,1,0.5 mg/mL) with different concentrations by taking physiological saline as a solvent, simultaneously setting distilled water and the physiological saline as positive and negative control groups respectively, placing all test tubes added with reagents into a constant-temperature water tank at 37 ℃ for water bath for 1h, centrifuging the samples at 4000rpm for 10 min, taking supernatant after centrifugation, measuring absorbance at 541 nm by using an ultraviolet spectrophotometer, and calculating according to the following formula:
hemolysis rate= (a Sample of -A Yin type vagina )/(A Yang (Yang) -A Yin type vagina )×100％
Wherein A is Sample of ，A Yang (Yang) ，A Yin type vagina The absorbance values of the sample to be measured, distilled water and physiological saline group are shown respectively.
Experimental results: the evaluation of the biological safety of the cationic nano-micelle prepared by the amphiphilic lipoic acid cationic chitosan derivative synthesized in the embodiment 1 is shown in figure 5, and the hemolysis rate of the cationic nano-micelle prepared by the amphiphilic compound synthesized in the embodiment 1 is lower than 5%, which indicates that the cationic nano-micelle has no obvious interference to erythrocytes and has good potential for in vivo application.
(4) Encapsulation efficiency and drug loading rate of cationic nano-micelles prepared by using the amphiphilic lipoyl cationic chitosan derivative synthesized in example 1 were determined.
The method comprises the following specific steps: 20 mg of the product of example 1 was weighed into dimethyl sulfoxide at a concentration of 2 mg/mL, and the solution was transferred into a dialysis bag (mwco=100), dialyzed against water at room temperature for 12 h, and kept ready for use after dialysis.
200 uL (15 mg/mL) of doxorubicin/dimethyl sulfoxide solution is taken, slowly dripped into the solution under strong stirring to obtain compound aqueous solution after dialysis, ultrasonic treatment is carried out for 20 min by a probe under the power condition of 135W, after ultrasonic treatment is finished, the obtained solution is centrifuged for 30 min at 12000rpm, the absorption degree of the supernatant is measured at 484 nm by an ultraviolet spectrophotometry, and the encapsulation efficiency and the drug loading rate of the drug-loaded nano-micelle are calculated according to the following formula.
Encapsulation efficiency = (total drug-free drug)/total drug x 100%
Drug loading = (total drug amount-free drug)/(total carrier + total drug) ×100%
Table 2, results of measuring encapsulation efficiency and drug loading of the cationic nano-micelle prepared in example 1
Experimental results: the encapsulation capacity of the amphiphilic lipoic acid acyl cationic chitosan derivative synthesized in the embodiment 1 of the invention for preparing the cationic nano micelle is shown in table 2, and the amphiphilic compound synthesized in the embodiment 1 has better encapsulation capacity for the chemotherapeutic drugs.
(5) Examination of drug-loaded nano-micelle in vitro drug release performance prepared by using amphiphilic lipoyl cationic chitosan derivative synthesized in example 1.
The method comprises the following specific steps: and (3) transferring a proper amount of the drug-loaded nanoparticle solution (about 0.7-mg containing doxorubicin) prepared in the step (4) into a dialysis bag (Mwco=8000-12000-Da), simultaneously taking an equal amount of doxorubicin dissolved in PBS solution as a control group, clamping two ends of the dialysis bag, placing the control group into a 150 mL buffer solution (PH=5.5), placing the drug-loaded nanoparticle into 150 mL release media (PH=5.5 buffer solution and PH=5.0+10 mmole GSH buffer solution) respectively, oscillating at 37 ℃ under the water bath condition of 100rpm, sampling 3mL at fixed time, supplementing an equal volume of release media, and measuring the absorbance of each sample at 484 nm.
Experimental results: the in-vitro drug release performance of the drug-loaded nano micelle prepared by the amphiphilic lipoic acid cationic chitosan derivative synthesized in the embodiment 1 of the invention is shown in figure 6, the free doxorubicin shows a sudden release phenomenon in a release medium, and 24 h is basically and completely released; compared with free doxorubicin, the release rate of doxorubicin entrapped in the nano-micelle prepared in example 1 is obviously reduced, and the phenomenon of "slow release" is presented; the release rate of the doxorubicin in the drug-loaded nano-micelle is obviously improved under the GSH condition of 10mM, which indicates that the nano-micelle prepared in the embodiment 1 has 'redox responsiveness', so that the doxorubicin can be rapidly released after reaching tumor tissues so as to kill tumor cells.
Claims (5)
1. An amphiphilic lipoyl cationic chitosan derivative, characterized in that: the structural formula of the amphiphilic lipoyl cationic chitosan derivative is shown as a formula (1),
formula (1), wherein
Wherein the value range of the polymerization degree n is 20-60.
2. A method for preparing the amphiphilic lipoyl cationic chitosan derivative according to claim 1, which is characterized in that: firstly, the chitosan is subjected to nicotinoylation, and then nucleophilic reaction with bromoethylamine hydrobromide to obtain aminoethyl cationic chitosan, and the obtained aminoethyl cationic chitosan is used in the following steps ofN,NReacting the'-carbonyl diimidazole with alpha-lipoic acid under the catalysis of the' -carbonyl diimidazole to obtain the amphiphilic lipoyl cationic chitosan derivative shown in the formula (1);
the saidN,NThe molar quantity of the' -carbonyl diimidazole is 3.0-6.0 times of that of the chitosan; the molar quantity of the nicotinic acid is 3.0-6.0 times of that of the chitosan; the molar quantity of the bromoethylamine hydrobromide is 2.0-5.0 times of that of the nicotinoylated chitosan;N,Nthe molar quantity of the' -carbonyl diimidazole is 1.0-4.0 times of the molar quantity of the alpha-lipoic acid; the molar quantity of the alpha-lipoic acid is 1.0 to 4.0 times of that of the aminoethyl cation chitosan.
3. The method for preparing the amphiphilic lipoyl cationic chitosan derivative according to claim 2, wherein the method comprises the following steps: will beN,NAdding' -carbonyl diimidazole into dimethyl sulfoxide solution containing nicotinic acid, then reacting with chitosan at 50-70 ℃ for 12-24 h, precipitating the reaction mixture with excessive acetone, washing, cooling and drying to obtain nicotinoylated chitosan; the obtained nicotinamide acylated chitosan and bromoethylamine hydrobromideReacting the acid salt at 50-70 ℃ for 12-36 h, precipitating the obtained solid with ethanol, washing, and freeze-drying to obtain the aminoethyl cation chitosan for later use.
4. The method for preparing the amphiphilic lipoyl cationic chitosan derivative according to claim 2, wherein the method comprises the following steps: will beN,N'-Adding carbonyl diimidazole into dimethyl sulfoxide solution containing alpha-lipoic acid, and reacting at 50-70 ℃ in nitrogen atmosphere for 12-36 h, wherein the obtained solution is prepared for standby; dissolving the obtained aminoethyl cationic chitosan in dimethyl sulfoxide, continuously reacting with the obtained solution at 50-70 ℃ for 12-36 h, then precipitating and washing with acetone, and freeze-drying to obtain the amphiphilic lipoyl cationic chitosan derivative shown in the formula (1).
5. Use of an amphiphilic lipoyl cationic chitosan derivative according to claim 1, characterized in that: the application of the nano micelle prepared by the amphiphilic lipoyl cationic chitosan derivative shown in the formula (1) in a drug carrier.
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| CN116496430A (en) * | 2023-06-26 | 2023-07-28 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan cystamine grafted derivative and preparation and application thereof |
| CN116589610A (en) * | 2023-07-17 | 2023-08-15 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan lipoic acid grafted derivative and preparation and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103945830A (en) * | 2011-02-25 | 2014-07-23 | 南达科他州立大学 | Polymer conjugated protein micelles |
| CN106581686A (en) * | 2016-12-15 | 2017-04-26 | 中国药科大学 | Preparation and application of hyaluronic acid-modified amphipathic chitosan derivative carrier with tumor microenvironment specificity drug release effect |
| CN108003354A (en) * | 2017-12-28 | 2018-05-08 | 浙江大学 | A kind of polymer and its preparation and application for responding intracellular acidic and redox environment |
| CN108420793A (en) * | 2017-12-26 | 2018-08-21 | 厦门本素药业有限公司 | A kind of blank mixed micelle and its preparation method and application |
| CN111658612A (en) * | 2020-06-10 | 2020-09-15 | 浙江工业大学 | Intelligent amphiphilic polymer nano micelle and preparation method and application thereof |
| CN114853922A (en) * | 2022-05-09 | 2022-08-05 | 中国科学院烟台海岸带研究所 | Thiocanyl chitosan quaternary ammonium salt and preparation method and application thereof |
-
2023
- 2023-05-04 CN CN202310486747.1A patent/CN116178589B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103945830A (en) * | 2011-02-25 | 2014-07-23 | 南达科他州立大学 | Polymer conjugated protein micelles |
| CN106581686A (en) * | 2016-12-15 | 2017-04-26 | 中国药科大学 | Preparation and application of hyaluronic acid-modified amphipathic chitosan derivative carrier with tumor microenvironment specificity drug release effect |
| CN108420793A (en) * | 2017-12-26 | 2018-08-21 | 厦门本素药业有限公司 | A kind of blank mixed micelle and its preparation method and application |
| CN108003354A (en) * | 2017-12-28 | 2018-05-08 | 浙江大学 | A kind of polymer and its preparation and application for responding intracellular acidic and redox environment |
| CN111658612A (en) * | 2020-06-10 | 2020-09-15 | 浙江工业大学 | Intelligent amphiphilic polymer nano micelle and preparation method and application thereof |
| CN114853922A (en) * | 2022-05-09 | 2022-08-05 | 中国科学院烟台海岸带研究所 | Thiocanyl chitosan quaternary ammonium salt and preparation method and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116496430A (en) * | 2023-06-26 | 2023-07-28 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan cystamine grafted derivative and preparation and application thereof |
| CN116496430B (en) * | 2023-06-26 | 2023-08-25 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan cystamine grafted derivative and preparation and application thereof |
| CN116589610A (en) * | 2023-07-17 | 2023-08-15 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan lipoic acid grafted derivative and preparation and application thereof |
| CN116589610B (en) * | 2023-07-17 | 2023-10-03 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan lipoic acid grafted derivative and preparation and application thereof |
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