CN113209028A - Insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles and preparation method thereof - Google Patents
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Abstract
The invention discloses an insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particle and a preparation method thereof. The phenylboronic acid group epsilon-polylysine is obtained by grafting carboxyl modified phenylboronic acid on epsilon-polylysine through a grafting reaction, and the mass part ratio of the carboxyl modified phenylboronic acid to the epsilon-polylysine is as follows: 1: (0.1-10) and then preparing the insulin-loaded epsilon-polylysine particles by soaking the epsilon-polylysine particles in an insulin solution. The preparation method is simple; the phenylboronic acid group epsilon-polylysine particles loaded with insulin have glucose response performance, and can automatically adjust the release rate and release dosage of the insulin; the insulin-loaded phenylboronate epsilon-polylysine particles can be used to deliver insulin by subcutaneous injection.
Description
Technical Field
The invention belongs to the technical field of high polymer materials, and particularly relates to insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles and a preparation method thereof.
Background
Diabetes is a chronic disease caused by loss or impairment of insulin secretion function, is mainly characterized by hyperglycemia, and can further induce functional impairment of heart, nerves, eyes, kidneys and the like in severe cases. According to the statistics of the international diabetes union, the number of diabetes patients in the whole world in 2019 is up to 4.63 hundred million. With the progress of the disease, the diabetic patients have a gradually increasing dependence on antidiabetic drugs, especially exogenous insulin. Currently, subcutaneous insulin injection is a common method for supplementing exogenous insulin for diabetics. However, the patient needs to adopt a strategy of injecting insulin several times a day to achieve satisfactory blood sugar control, and thus the burden on the patient is large.
Insulin-loaded glycophenylboronic acid-based polymer particles have recently attracted considerable attention in the field of diabetes therapy. The principle is that insulin is loaded in glucose responsive polymer particles containing hydrophobic phenylboronic acid groups through hydrophobic action, and after the insulin loaded phenylboronic acid based polymer particles are delivered into a diabetic patient, because the hydrophobic phenylboronic acid groups can be combined with glucose into hydrophilic glucose/phenylboronic acid complex groups, the glucose can induce the dissolution of the phenylboronic acid based polymer particles, and then the loaded insulin is released. The higher the blood sugar concentration is, the more insulin is released, and the more remarkable the blood sugar reducing effect is; after the blood sugar concentration is reduced, the insulin release amount is correspondingly reduced, and the unreleased insulin can be stored and released when the blood sugar is increased next time. Based on the glucose-responsive insulin delivery properties described above, the insulin-loaded phenylboronic acid-based polymer particles can control blood glucose levels to normal levels while reducing injection frequency and avoiding the pain of multiple injections a day. At present, the phenylboronic acid-based polymer particles have the problems of complex synthesis process and high large-scale production cost. For example, the synthesis of polyethylene glycol modified phenylboronic acid-based polymer particles reported by Li et al requires at least 5 steps; the synthesis of starch modified phenylboronic acid based polymer particles reported by Wen et al required 3 steps.
Disclosure of Invention
In order to solve the problems, the invention provides insulin-loaded glycophenborate epsilon-polylysine particles. The phenylboronic acid group epsilon-polylysine particles are prepared by adopting a grafting reaction, and the phenylboronic acid group epsilon-polylysine particles are soaked in an insulin solution to prepare the insulin-loaded phenylboronic acid group epsilon-polylysine particles. The preparation method of the sugar-sensitive phenylboronic acid group epsilon-polylysine disclosed by the invention is simple and only needs one-step synthesis reaction; the phenylboronic acid group epsilon-polylysine particles loaded with insulin have glucose response performance, and can automatically adjust the release rate and the release dosage of the insulin.
The technical scheme adopted by the invention is as follows:
insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles
The phenylboronic acid group epsilon-polylysine is obtained by grafting carboxyl modified phenylboronic acid on epsilon-polylysine through a grafting reaction, and the mass part ratio of the carboxyl modified phenylboronic acid to the epsilon-polylysine is as follows: 1: (0.1-10).
The structural formula of epsilon-polylysine is as follows:
the structural formula of the phenylboronic acid group epsilon-polylysine is as follows:
r is a structural formula of phenylboronic acid group epsilon-polylysine, R is one or more of four groups shown below, and the grafting ratio of the phenylboronic acid group is y/x.
Preparation method of insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles
The method comprises the following steps:
1) adding carboxyl modified phenylboronic acid and auxiliary materials into a solvent, and stirring at the stirring speed of 0-1500rpm for 0-240min at normal temperature to obtain a mixture A;
2) adding epsilon-polylysine into a solvent, and stirring at normal temperature under the same stirring condition as that in the step 1) to obtain a mixture B;
3) mixing the mixture A and the mixture B, and stirring at the stirring speed of 0-1500rpm for 5-3000min at normal temperature to obtain a mixture C;
4) putting the mixture C into a dialysis bag with the molecular weight cutoff of 500-;
5) soaking the epsilon-polylysine in an insulin solution at normal temperature for 10-240min, centrifuging at the rotation speed of 100 plus 10000rpm for 1-60min, removing supernatant, collecting solids, and freeze-drying to obtain the insulin-loaded glucose-sensitive epsilon-polylysine particles.
In the step 1), the auxiliary material is one or more of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide.
In the step 1), the carboxyl-modified phenylboronic acid comprises one or more of 2-carboxyl phenylboronic acid, 4-carboxyl-3-chlorobenzene boronic acid, 4-carboxyl-3-fluorobenzeneboronic acid and 4-carboxyl-2-chlorobenzene boronic acid.
The solvent in the step 1) and the step 2) is one or more of water, dimethyl sulfoxide, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dichloromethane and chloroform.
In the step 1), by mass, 1 part of carboxyl modified phenylboronic acid, 0.2-10 parts of auxiliary materials and 10-300 parts of solvent are used.
In the step 2), the epsilon-polylysine accounts for 1 part by mass, and the solvent accounts for 1-100 parts by mass.
The carboxyl modified phenylboronic acid in the step 1) and the epsilon-polylysine in the step 2) are counted by mass parts, wherein the carboxyl modified phenylboronic acid is 1 part, and the epsilon-polylysine is 0.1-10 parts.
In the step 5), the phenylboronic acid group epsilon-polylysine particles and the insulin are counted in parts by mass as 1 part and 0.01-1 part respectively.
The solvent of the insulin solution in the step 5) is water, the insulin is one or more of porcine insulin and human insulin, and the mass concentration of the insulin is 0.01-10%.
The phenylboronic acid group epsilon-polylysine particles are loaded with insulin and then delivered by a subcutaneous injection method.
The steps 1) to 3) can also be combined into one step, namely, the carboxyl modified phenylboronic acid, the auxiliary material, the epsilon-polylysine and the solvent are mixed and stirred to obtain a mixture C. Wherein the carboxyl modified phenylboronic acid comprises one or more of 2-carboxyl phenylboronic acid, 4-carboxyl-3-chlorobenzene boronic acid, 4-carboxyl-3-fluorobenzeneboronic acid and 4-carboxyl-2-chlorobenzene boronic acid. The auxiliary material comprises one or more of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. The solvent comprises one or more of water, dimethyl sulfoxide, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dichloromethane and chloroform. By mass, 1 part of carboxyl modified phenylboronic acid, 0.2-10 parts of auxiliary materials, 0.1-10 parts of epsilon-polylysine and 10-300 parts of solvent.
In the insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles, epsilon-polylysine is modified by hydrophobic phenylboronic acid and self-assembled to form phenylboronic acid group epsilon-polylysine particles, and insulin can be loaded in the particles through hydrophobic action. Because the hydrophobic phenylboronic acid group can be combined with glucose to form a hydrophilic glucose/phenylboronic acid compound group, the glucose can induce the dissolution of the phenylboronic acid polymer particles, and then the loaded insulin is released. After the phenylboronic acid-based polymer particles loaded with insulin are delivered into a diabetic patient, the higher the blood sugar concentration of the patient is, the more insulin is released by the phenylboronic acid-based polymer particles loaded with insulin, and the more remarkable the blood sugar reduction effect is; after the blood sugar concentration is reduced, the insulin release amount is correspondingly reduced, and the insulin which is not released can be stored and loaded on the glucose-sensitive phenylboronic acid group epsilon-polylysine particles until the next blood sugar is increased.
The invention has the beneficial effects that:
1. the synthesis process of the phenylboronic acid group epsilon-polylysine is simple, the synthesis reaction only needs 1 step, and the large-scale production cost is low.
2. For the glucose-sensitive phenylboronic acid group epsilon-polylysine particles loaded with insulin, the insulin loading process is mild, and the problem of insulin inactivation is avoided.
3. The phenylboronic acid group epsilon-polylysine particles can be completely hydrolyzed into lysine and carboxyl modified phenylboronic acid micromolecules, and the excretion of the phenylboronic acid group epsilon-polylysine particles in vivo is facilitated.
4. The glucose-sensitive phenylboronic acid group epsilon-polylysine particles loaded with insulin have stimulation-response performance on glucose concentration, and the release rate and the release dosage of the loaded drug can be automatically adjusted according to the change of the glucose concentration.
Summarizing, the preparation method of the invention is simple; the problem of insulin inactivation is not caused, and the prepared phenylboronic acid group epsilon-polylysine particles loaded with insulin have glucose response performance and can automatically adjust the release rate and release dosage of the insulin; and the phenylboronate epsilon-polylysine particles loaded with insulin can deliver insulin by subcutaneous injection.
Drawings
FIG. 1 is a nuclear magnetic hydrogen spectrum of phenylboronic acid group ε -polylysine prepared in example 1;
FIG. 2 is a graph of the cumulative release of insulin from insulin-loaded glycophenylboronate ε -polylysine particles prepared in example 9;
FIG. 3 is a circular dichroism plot of insulin for insulin-loaded glycophenylboronate ε -polylysine particles prepared in example 9.
Detailed Description
The present invention is described in more detail below with reference to examples, but the present invention is not limited thereto, and those skilled in the art can make various modifications and improvements without departing from the principle of the present invention, and the modifications and improvements are also considered to be within the scope of the present invention. Those not described in detail in this specification are within the skill of the art.
The embodiment of the invention is as follows:
example 1:
1.0780g of 4-carboxy-3-fluorobenzeneboronic acid, 1.1245g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 0.6739g N-hydroxysuccinimide are put into 100mL of dimethyl sulfoxide and stirred to obtain a mixture A, wherein the stirring speed is 400rpm, the stirring time is 90min, and the stirring temperature is normal temperature. And (3) putting 1.0001g of epsilon-polylysine into 10mL of water, and stirring at the speed of 400rpm for 10min at the normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at the stirring temperature of 30 ℃ to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 9 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
The results of this example are shown in FIG. 1, where FIG. 1 is the nuclear magnetic hydrogen spectrum of phenylboronic acid group ε -polylysine. The calculated graft ratio of phenylboronic acid groups was 65%. The synthesis process of the phenylboronic acid group epsilon-polylysine is simple, the synthesis reaction only needs 1 step, and the large-scale production cost is low.
Example 2:
1.3345g of 2-carboxyphenylboronic acid and 1.5087g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride were added to 50mL of water and 150mL of N, N-dimethylformamide to obtain a mixture A, and stirring was performed at 0rpm for 0min at room temperature. 1.2010g of epsilon-polylysine is put into 10mL of water and 2mL of N, N-dimethylformamide to be stirred, the stirring speed is 400rpm, the stirring time is 5min, and the stirring temperature is normal temperature, so that a mixture B is obtained. And mixing and stirring the mixture A and the mixture B at the stirring speed of 300rpm for 300min at the stirring temperature of normal temperature to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 3500Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 8h every time, wherein the water changing times are 12 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 3:
1.0001g of 4-carboxy-3-chlorobenzeneboronic acid, 0.9122g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 0.7397g N-hydroxysuccinimide were added to 50mL of water and 150mL of N, N-dimethylacetamide to obtain a mixture A, and the stirring rate was 200rpm, the stirring time was 60min, and the stirring temperature was room temperature. 1.0212g of epsilon-polylysine is put into 15mL of water and 1mL of N, N-dimethylacetamide and stirred at the stirring speed of 200rpm for 15min at the normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 200rpm for 1200min at the stirring temperature of normal temperature to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 500Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 8 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 4:
1.0005g of 4-carboxy-2-chlorobenzeneboronic acid, 1.5451g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.3125g N-hydroxysuccinimide were put into 50mL of water and 150mL of tetrahydrofuran to obtain a mixture A, and the stirring rate was 500rpm, the stirring time was 120min, and the stirring temperature was room temperature. 1.0012g of epsilon-polylysine was put into 9mL of water and 3mL of tetrahydrofuran and stirred at a stirring speed of 500rpm for 120min at a normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 120min at the stirring temperature of normal temperature to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 8 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 5:
1.0004g of 4-carboxy-3-chlorobenzeneboronic acid, 1.0014g of 4-carboxy-2-chlorobenzeneboronic acid, 2.2534g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.7533g N-hydroxysuccinimide are put into 100mL of dimethyl sulfoxide, 30mL of dichloromethane and 30mL of trichloromethane to obtain a mixture A, the stirring speed is 500rpm, the stirring time is 120min, and the stirring temperature is normal temperature. 1.1012g of epsilon-polylysine is put into 15mL of water, 3mL of dimethyl sulfoxide, 3mL of dichloromethane and 3mL of chloroform to be stirred, the stirring speed is 500rpm, the stirring time is 120min, and the stirring temperature is normal temperature, so that a mixture B is obtained. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 120min at the stirring temperature of normal temperature to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 8 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 6:
1.0780g of 4-carboxy-3-fluorobenzeneboronic acid, 1.1245g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.0001g of epsilon-polylysine are put into 100mL of water to be mixed and stirred, the stirring speed is 500rpm, the stirring time is 2000min, and the stirring temperature is normal temperature, so that a mixture C is obtained. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 9 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 7:
1.0006g of 2-carboxyphenylboronic acid, 1.0002g of 4-carboxy-3-chlorobenzeneboronic acid, 2.2242g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 2.0001g N-hydroxysuccinimide and 2.0002g of epsilon-polylysine were added to 100mL of water, 50mL of N, N-dimethylformamide, 50mL of N, N-dimethylacetamide and 50mL of tetrahydrofuran, and the mixture was stirred at 800rpm for 1000min at room temperature to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 9 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 8:
1.0021g of 4-carboxy-2-chlorobenzeneboronic acid, 1.3459g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.0008g of epsilon-polylysine were put into 100mL of water, 50mL of dimethyl sulfoxide, 50mL of dichloromethane and 50mL of chloroform and mixed and stirred at the stirring speed of 800rpm for 1000min at normal temperature to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 9 times. And after dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product of phenylboronic acid group epsilon-polylysine.
Example 9:
10.0mg of the product, phenylboronyl epsilon-polylysine, obtained in example 1 was soaked in 1mL of a 0.128% aqueous solution of human insulin for 60min at room temperature with a stirring rate of 250 rpm. Then, the mixture was centrifuged at 3000rpm for 20 min. The supernatant was removed, the solids were collected and lyophilized to obtain insulin-loaded glycophenylboronate epsilon-polylysine particles with an insulin load of 0.102(mg insulin)/(mg insulin-loaded phenylboronate epsilon-polylysine particles).
The results of this embodiment are shown in FIGS. 2-3. FIG. 2 is a graph of the cumulative release of insulin from insulin-loaded particles of glycophenylboronate ε -polylysine, using a glucose concentration of 4g/L to simulate high blood glucose levels, a glucose concentration of 1g/L to simulate normal blood glucose levels, and a glucose concentration of 0g/L as a reference. The higher the glucose concentration, the more insulin is released. Fig. 3 is a circular dichroism chart of insulin of the sugar-sensitive phenylboronic acid group epsilon-polylysine particles loaded with insulin, and the result shows that the secondary structure of the insulin loaded on the sugar-sensitive phenylboronic acid group epsilon-polylysine particles is not different from that of natural insulin, which shows that the insulin loading process of the sugar-sensitive phenylboronic acid group epsilon-polylysine particles is mild and does not generate the problem of insulin inactivation. 0.55mg of the insulin-loaded glycopyrrolate epsilon-polylysine particles are injected into a diabetic SD rat body with the weight of 200g, the hyperglycemia level of 5g/L can be reduced to the normal blood sugar level of 0.5-2g/L within 0.5 hour, and then the normal blood sugar level can be maintained for more than 9 hours. 1mg of the granules of the sugar-sensitive phenylboronic acid group epsilon-polylysine are injected into a diabetic SD rat body with the weight of 200g, and can be completely excreted within 24 hours.
Claims (10)
1. An insulin-loaded glycophenborate epsilon-polylysine particle, which is characterized in that: the phenylboronic acid group epsilon-polylysine is obtained by grafting carboxyl modified phenylboronic acid on epsilon-polylysine by using a grafting reaction.
2. A method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 1, comprising: the method comprises the following steps:
1) adding carboxyl modified phenylboronic acid and auxiliary materials into a solvent, and stirring at the stirring speed of 1-1500rpm for 1-240min at normal temperature to obtain a mixture A;
2) adding epsilon-polylysine into a solvent, and stirring at normal temperature under the same stirring condition as that in the step 1) to obtain a mixture B;
3) mixing the mixture A and the mixture B, and stirring at the stirring speed of 1-1500rpm for 5-3000min at normal temperature to obtain a mixture C;
4) putting the mixture C into a dialysis bag, placing the bag in water for dialysis, changing the dialysis water once every 2-24 hours, and freeze-drying the mixture in the dialysis bag after the dialysis is finished to obtain a product of phenylboronic acid group epsilon-polylysine;
5) soaking the epsilon-polylysine in an insulin solution at normal temperature for 10-240min, centrifuging at the rotation speed of 100 plus 10000rpm for 1-60min, removing supernatant, collecting solids, and freeze-drying to obtain the insulin-loaded glucose-sensitive epsilon-polylysine particles.
3. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: in the step 1), the auxiliary material is one or more of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide.
4. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: in the step 1), the carboxyl-modified phenylboronic acid comprises one or more of 2-carboxyl phenylboronic acid, 4-carboxyl-3-chlorobenzene boronic acid, 4-carboxyl-3-fluorobenzeneboronic acid and 4-carboxyl-2-chlorobenzene boronic acid.
5. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: the solvent in the step 1) and the step 2) is one or more of water, dimethyl sulfoxide, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dichloromethane and chloroform.
6. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: in the step 1), by mass, 1 part of carboxyl modified phenylboronic acid, 0.2-10 parts of auxiliary materials and 10-300 parts of solvent are used.
7. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: in the step 2), the epsilon-polylysine accounts for 1 part by mass, and the solvent accounts for 1-100 parts by mass.
8. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: the carboxyl modified phenylboronic acid in the step 1) and the epsilon-polylysine in the step 2) are counted by mass parts, wherein the carboxyl modified phenylboronic acid is 1 part, and the epsilon-polylysine is 0.1-10 parts.
9. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: in the step 5), the phenylboronic acid group epsilon-polylysine particles and the insulin are counted in parts by mass as 1 part and 0.01-1 part respectively.
10. The method of preparing insulin-loaded glycopyrrolate epsilon-polylysine particles of claim 2, wherein: the solvent of the insulin solution in the step 5) is water, the insulin is one or more of porcine insulin and human insulin, and the mass concentration of the insulin is 0.01-10%.
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| Application Number | Priority Date | Filing Date | Title |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114632145A (en) * | 2022-03-07 | 2022-06-17 | 浙江大学 | Insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and preparation method thereof |
| WO2024067600A1 (en) * | 2022-09-29 | 2024-04-04 | 浙江大学 | Glucose-responsive complex, preparation method therefor, and use thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111763317A (en) * | 2019-03-13 | 2020-10-13 | 华东师范大学 | Polymer material modified by phenylboronic acid and application thereof in intracellular delivery of protein and polypeptide |
-
2021
- 2021-03-31 CN CN202110346422.4A patent/CN113209028A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111763317A (en) * | 2019-03-13 | 2020-10-13 | 华东师范大学 | Polymer material modified by phenylboronic acid and application thereof in intracellular delivery of protein and polypeptide |
Non-Patent Citations (2)
| Title |
|---|
| 吕娟等: "含苯硼酸材料在药物投递中的研究进展", 《科学通报》 * |
| 罗磊: "负载胰岛素的壳聚糖-苯硼酸接枝物的制备及体外释放", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114632145A (en) * | 2022-03-07 | 2022-06-17 | 浙江大学 | Insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and preparation method thereof |
| CN114632145B (en) * | 2022-03-07 | 2024-02-27 | 浙江大学 | Insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and preparation method thereof |
| WO2024067600A1 (en) * | 2022-09-29 | 2024-04-04 | 浙江大学 | Glucose-responsive complex, preparation method therefor, and use thereof |
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