Intelligent dual-responsive drug-loaded myocardial injection hydrogel and preparation method thereof

Abstract

The application discloses an intelligent double-response drug-loaded myocardial injection hydrogel and a preparation method thereof, and belongs to the field of biomedical materials. In order to solve the problem of realizing the accurate release of drugs in time and space according to the changes of microenvironments after ischemia and different conditions of diseases, N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, then a certain volume of dimethylformamide solution is added, and the mixture is stirred and reacted under oil bath conditions for 0.5h-4h, the mixture is added into a certain volume of ice tetrahydrofuran, and then is placed into a refrigerator at-20 DEG C and is left for 10h-12h, white solid is separated out, centrifugal separation and purification treatment are carried out, and then vacuum freeze-drying treatment is carried out to obtain (3,3'-p-boronic acid phenylmethylammonium bromide)-methyl dipropylamine, a drug-loaded aqueous solution is prepared, polyvinyl alcohol solution is added, and then mixing and rapid stirring are carried out for 1min-2min, so as to obtain the intelligent double-response drug-loaded myocardial injection hydrogel.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of biomedical materials, and particularly relates to an intelligent double-response drug-loaded myocardial injection hydrogel and a preparation method thereof. BACKGROUND

[0002] At present, the irreversible structural damage, abnormal microenvironment and microcirculation damage of the heart caused by myocardial ischemia are difficult to improve by traditional stents and bypass techniques. At this time, the tissue engineering method plays a unique advantage. The mechanical properties of the myocardial patch and the hydrogel itself can make up for the stress difference of the ischemic area of the heart, and the different drugs loaded in the patch and the hydrogel can improve the microenvironment of the ischemic area to reduce the loss of myocardial cells. However, the current patch and hydrogel carrier has certain limitations. The drug release of the patch and the hydrogel carrier only relies on the degradability of the material itself, and cannot achieve precise drug release for local lesions. At the same time, due to the inhomogeneity of the acid change and the active oxygen change after myocardial ischemia, the accuracy of the single-sensitive carrier to identify the lesion is poor, thereby affecting the treatment effect of the drug on the local lesion. Moreover, in the method of responding to two different environmental conditions by two chemical bonds, the product is not easy to be clinically transformed due to the complexity of the reaction process and conditions. SUMMARY

[0003] The problems to be solved by the application are to sense the change of the microenvironment after ischemia and to achieve the precise release of the drug in time and space according to different conditions of the lesion, and an intelligent double-response drug-loaded myocardial injection hydrogel and a preparation method thereof are provided.

[0004] To achieve the above-mentioned purpose, the application realizes the following technical scheme:

[0005] A preparation method of an intelligent double-response drug-loaded myocardial injection hydrogel, comprising the following steps:

[0006] S1. A certain amount of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, and then a certain volume of dimethylformamide solution is added, and the mixture is stirred and reacted under oil bath conditions for 0.5h-4h to obtain a mixture, which is used for standby;

[0007] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1 according to the volume ratio, and then placed in a-20℃ refrigerator for 10h-12h, and a white solid is precipitated, which is used for standby;

[0008] S3. The white solid obtained in step S2 is added to a certain volume of ice tetrahydrofuran, and 2-3 times of centrifugal separation purification treatment is carried out, and then vacuum freeze-drying treatment is carried out to obtain (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine, which is used for standby;

[0009] S4. A certain amount of drug-loaded medicine is weighed and added to ultrapure water to obtain a drug-loaded medicine aqueous solution;

[0010] S5. The drug-loaded medicine aqueous solution obtained in step S4 is added to (3,3'-p-boronic acid phenylmethyl ammonium bromide)-methyl dipropylamine obtained in step S3 according to the mass ratio, and a certain volume of polyvinyl alcohol solution is added after uniform mixing, mixed and rapidly stirred for 1-2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0011] Further, the mass of N'N-bis(3-aminopropyl)methylamine in step S1 is 0.11 g, the mass of 4-(bromomethyl)phenylboronic acid is 0.333 g, the volume of the dimethylformamide solution is 15 ml, and the concentration of the dimethylformamide solution is 98 wt%.

[0012] Further, the oil bath temperature in step S1 is 25-50 DEG C, and the stirring speed is 200-500 rpm / min.

[0013] Further, the volume ratio of the mixture to tetrahydrofuran in step S2 is 10-20:35-50, the temperature of the ice to tetrahydrofuran is -20 DEG C, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1.

[0014] Further, the centrifugal separation and purification treatment in step S3 is carried out at 4 DEG C at a speed of 4000-4500 rpm for 4-6 min, and the vacuum freeze-drying treatment time is 10-14 h.

[0015] Further, the drug-loaded medicine in step S4 is glutamine, and the ratio of the drug-loaded medicine to ultrapure water is 0.22 g:10 ml.

[0016] Further, in step S5, the drug-loaded medicine aqueous solution is dissolved with (3,3'-p-boronic acid phenylmethyl ammonium bromide)-methyl dipropylamine at a proportion of 1.5% W / W, and after uniform mixing, polyvinyl alcohol solution is added according to a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 3.5% W / W-4.5% W / W, and the mixture is mixed and rapidly stirred for 1-2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0017] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel is a kind of intelligent dual-responsive drug-loaded myocardial injection hydrogel, and the intelligent dual-responsive drug-loaded myocardial injection hydrogel is a dual-sensitive drug release hydrogel carrier.

[0018] The beneficial effects of the present application are:

[0019] The preparation method of the intelligent double-response drug-loaded myocardial injection hydrogel provided by the application, the intelligent drug carrier mainly senses the local microenvironment changes of the lesion, and when myocardial ischemia occurs, the local microenvironment in the area will have different degrees of increase in acidity and active oxygen, and the drug release of the intelligent drug release carrier of the myocardium mainly relies on the chemical bond rupture after sensing the local acidity and active oxygen, so that the drug is released as needed. Therefore, the chemical bond capable of sensing acidity and active oxygen needs to be included in the synthesis process of the material, or two chemical bonds respond to different conditions respectively, or the same chemical bond responds to two conditions. In the design of the application, a chemical bond capable of sensing acidity and active oxygen, borate ester bond, is found and the design is carried out around this chemical bond. First, N'N-bis(3-aminopropyl)methylamine is combined with 4-bromomethylphenylboronic acid to obtain a new boronic acid-containing compound, (3,3'-p-boronic acid phenylmethyl ammonium bromide)-methyl dipropylamine (BBMDP). Then, the drug is dissolved in ultrapure water to obtain a mixed solution. Finally, (3,3'-p-boronic acid phenylmethyl ammonium bromide)-methyl dipropylamine is purified and dissolved in the mixed solution containing glutamine, and then mixed with polyvinyl alcohol in proportion, and quickly stirred to obtain a double-sensitive drug-loaded hydrogel containing borate ester bond. The preparation method is simple and feasible, and is easy to be clinically transformed.

[0020] The intelligent double-response drug-loaded myocardial injection hydrogel provided by the application can sense two main changes of the microenvironment after ischemia, and achieve accurate release of the drug in time and space and release as needed according to different conditions of the lesion. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 It is a structural schematic diagram of (3,3'-p-boronic acid phenylmethyl ammonium bromide)-methyl dipropylamine provided by the application;

[0022] Figure 2 It is a nuclear magnetic hydrogen spectrum of (3,3'-p-boronic acid phenylmethyl ammonium bromide)-methyl dipropylamine provided by the application;

[0023] Figure 3 It is a microstructure photo of the intelligent double-response drug-loaded myocardial injection hydrogel provided by the application under a freeze scanning electron microscope;

[0024] Figure 4 It is a sensitivity comparison curve of active oxygen of the intelligent double-response drug-loaded myocardial injection hydrogel provided by the application;

[0025] Figure 5 It is a sensitivity comparison curve of acidic environment of the intelligent double-response drug-loaded myocardial injection hydrogel provided by the application;

[0026] Figure 6Comparison chart of residual mass of the product prepared for the first to fourth embodiments of the present application at pH 5.0 within five consecutive days per day;

[0027] Figure 7 Comparison chart of residual mass of the product prepared for the first to fourth embodiments of the present application at 0.1 mM H2O2 within four consecutive days per day. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solutions and advantages of the present application more clear, the present application is further described in detail below in combination with the drawings and specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application, i.e., the described specific embodiments are only a part of the embodiments of the present application, but not all the specific embodiments. The components of the specific embodiments of the present application generally described and shown in the drawings herein can be arranged and designed in various different configurations, and the present application can also have other embodiments.

[0029] Therefore, the detailed description of the specific embodiments of the present application provided below in the drawings is not intended to limit the scope of the claimed present application, but only represents selected specific embodiments of the present application. All other specific embodiments obtained by those skilled in the art based on the specific embodiments of the present application without making creative efforts, fall within the scope of the present application.

[0030] In order to further understand the invention content, characteristics and effects of the present application, the following specific embodiments are exemplified, and the drawings are attached Figure 1 - the drawings Figure 7 The detailed description is as follows: Specific embodiment one:

[0032] A preparation method of an intelligent dual-response drug-loaded myocardial injection hydrogel, comprising the following steps:

[0033] S1. A certain mass of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, and then a certain volume of dimethylformamide solution is added, and the mixture is stirred under oil bath conditions for 2h to obtain a mixture, which is ready for use;

[0034] Further, in step S1, the mass of N'N-bis(3-aminopropyl)methylamine is 0.11g, the mass of 4-(bromomethyl)phenylboronic acid is 0.333g, the volume of the dimethylformamide solution is 15ml, and the concentration of the dimethylformamide solution is 98wt%;

[0035] Further, in step S1, the oil bath temperature is 40℃, and the stirring speed is 200rpm;

[0036] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1, and then placed in a refrigerator at -20°C for 12h to precipitate white solids, which are used as is;

[0037] Further, the volume ratio of the mixture in step S2 to tetrahydrofuran is 15:40, the temperature of the ice tetrahydrofuran is -20°C, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1;

[0038] S3. The white solids obtained in step S2 are added to a certain volume of ice tetrahydrofuran, and then subjected to two centrifugal separation purification treatments, followed by vacuum freeze-drying treatment to obtain (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine, which is used as is;

[0039] Further, the centrifugal separation purification treatment in step S3 is performed at 4°C at a speed of 4000rpm for 5min, and the vacuum freeze-drying treatment is performed for 12h;

[0040] S4. A certain mass of drug-loaded is weighed and added to ultrapure water to obtain a drug-loaded aqueous solution, which is used as is;

[0041] Further, the drug-loaded in step S4 is glutamine, and the mass ratio of the drug-loaded to the ultrapure water is 0.22g:10ml;

[0042] S5. The drug-loaded aqueous solution obtained in step S4 is mixed with (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine obtained in step S3 according to the mass ratio, and then a certain volume of polyvinyl alcohol solution is added, mixed and stirred rapidly for 2min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0043] Further, in step S5, the drug-loaded aqueous solution is dissolved with (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine at a proportion of 1.5%W / W, and then polyvinyl alcohol solution is added according to a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 4%W / W, and the mixture is mixed and stirred rapidly for 1min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0044] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment of the application can prepare an intelligent dual-responsive drug-loaded myocardial injection hydrogel, which is a dual-sensitive drug release hydrogel carrier. Figure 2 NMR is used to determine the molecular structure of the new cross-linking agent;

[0045] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment prepares an intelligent dual-responsive drug-loaded myocardial injection hydrogel. The obtained intelligent dual-responsive drug-loaded myocardial injection hydrogel sample is subjected to 24 hours of freeze-drying treatment, then the sample is quickly frozen with liquid nitrogen to make it fragile, so as to break the cross section; then, the sample is fixed on an observation platform using a conductive adhesive, and the cross section is ensured to face upwards, and then vacuum sputtering gold is performed to increase the conductivity; after the above steps are completed, the sample is placed into an electron microscope, and clear images are selected under different magnifications for shooting and collecting relevant data. The scanning electron microscope photos of the injectable hydrogel in the obtained hydrogel composition are as shown in Figure 3 It can be seen that it is a three-dimensional, porous sample structure. Figure 3

[0046] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment prepares an intelligent dual-responsive drug-loaded myocardial injection hydrogel. In order to verify the ROS sensitivity of the hydrogel, the hydrogel patch is added into a non-active oxygen group (PBS group) and a H2O2 solution containing 0.005 mmol / L, 0.01 mmol / L and 0.05 mmol / L respectively, and the mass change of the hydrogel patch is recorded at different time points, for a total of 60 hours. The ROS responsiveness curve of the hydrogel patch in the hydrogel composition is as shown in Figure 4 It can be seen from the figure that the degradation rate is the fastest when the concentration of H2O2 is 0.05 mmol / L, and the hydrogel composition has good ROS sensitivity.

[0047] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment prepares an intelligent dual-responsive drug-loaded myocardial injection hydrogel. In order to verify the pH sensitivity of the hydrogel, the hydrogel patch is added into a PBS solution with pH=7.4, pH=6.5 and pH=5.0 respectively, and the mass change of the hydrogel patch is recorded every day, for a total of 18 days. The pH sensitivity curve of the hydrogel patch in the hydrogel composition is as shown in Figure 5 The sensitivity of the hydrogel to active oxygen environment and acidic environment is reflected by the statistical daily weight remaining percentage. The statistical results show that the degradation rate gradient of the hydrogel increases with the decrease of pH, which indicates that the hydrogel has good pH sensitivity. Specific embodiment two

[0049] A preparation method of an intelligent dual-responsive drug-loaded myocardial injection hydrogel, comprising the following steps:

[0050] S1. A certain amount of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, and then a certain volume of dimethylformamide solution is added. The mixture is stirred and reacted under oil bath conditions for 0.5 h, and is ready for use.​

[0051] Further, the mass of N'N-bis(3-aminopropyl)methylamine in step S1 is 0.11 g, the mass of 4-(bromomethyl)phenylboronic acid is 0.333 g, the volume of dimethylformamide solution is 15 ml, and the concentration of dimethylformamide solution is 98 wt%;

[0052] Further, the oil bath temperature in step S1 is 25-50°C, and the stirring speed is 300 rpm;

[0053] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1, and then placed in a-20°C refrigerator for 10 h to precipitate white solids, which are used as needed;

[0054] Further, the volume ratio of the mixture and tetrahydrofuran in step S2 is 10:35, the temperature of ice tetrahydrofuran is-20°C, and the volume ratio of tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1;

[0055] S3. The white solids obtained in step S2 are added to a certain volume of ice tetrahydrofuran, and then subjected to 2 times of centrifugal separation and purification treatment, followed by vacuum freeze-drying treatment to obtain (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine, which is used as needed;

[0056] Further, the centrifugal separation and purification treatment in step S3 is carried out at 4°C at a speed of 4000 rpm for 4 min, and the vacuum freeze-drying treatment is carried out for 10 h;

[0057] S4. A certain mass of drug-loaded is weighed and added to ultrapure water to obtain a drug-loaded aqueous solution, which is used as needed;

[0058] Further, the drug-loaded in step S4 is glutamine, and the mass ratio of drug-loaded to ultrapure water is 0.22 g:10 ml;

[0059] S5. The drug-loaded aqueous solution obtained in step S4 is added with (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine obtained in step S3 according to the mass ratio, mixed uniformly, and then added with a certain volume of polyvinyl alcohol solution, mixed and stirred rapidly for 1 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0060] Further, in step S5, the drug-loaded aqueous solution is dissolved with (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine at a proportion of 1.5% W / W, mixed uniformly, and then added with polyvinyl alcohol solution according to a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 3.5 W / W, mixed and stirred rapidly for 1 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0061] The preparation method of the intelligent dual-response drug-loaded myocardial injection hydrogel is used for preparing an intelligent dual-response drug-loaded myocardial injection hydrogel. Specific embodiment three

[0063] The preparation method of the intelligent dual-response drug-loaded myocardial injection hydrogel comprises the following steps:

[0064] S1. A certain mass of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, and then a certain volume of dimethylformamide solution is added, and stirring is carried out under oil bath conditions for 4 h to obtain a mixture, which is used as needed.

[0065] Further, in step S1, the mass of N'N-bis(3-aminopropyl)methylamine is 0.11 g, the mass of 4-(bromomethyl)phenylboronic acid is 0.333 g, the volume of the dimethylformamide solution is 15 ml, and the concentration of the dimethylformamide solution is 98 wt%.

[0066] Further, in step S1, the oil bath temperature is 50°C, and the stirring speed is 500 rpm.

[0067] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1 according to a volume ratio, and then placed in a-20°C refrigerator for 12 h to precipitate white solids, which are used as needed.

[0068] Further, in step S2, the volume ratio of the mixture to tetrahydrofuran is 20:50, the temperature of the ice tetrahydrofuran is-20°C, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1.

[0069] S3. The white solids obtained in step S2 are added to a certain volume of ice tetrahydrofuran, and then subjected to 3 times of centrifugal separation and purification treatment, and then subjected to vacuum freeze-drying treatment to obtain (3,3'-p-boronic acid phenyl methyl ammonium bromide)-methyl dipropylamine, which is used as needed.

[0070] Further, in step S3, the centrifugal separation and purification treatment is carried out at 4°C at a speed of 4500 rpm for 6 min, and the vacuum freeze-drying treatment is carried out for 14 h.

[0071] S4. A certain mass of a drug is weighed and added to ultrapure water to obtain a drug-loaded aqueous solution, which is used as needed.

[0072] Further, in step S4, the drug is glutamine, and the mass ratio of the drug to ultrapure water is 0.22 g:10 ml.

[0073] S5. The drug-loaded aqueous solution obtained in step S4 is added with (3,3'-p-boronic acid phenyl methyl ammonium bromide)-methyl dipropylamine in a mass ratio, mixed uniformly, and then a certain volume of polyvinyl alcohol solution is added, mixed and stirred rapidly for 2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0074] Further, in step S5, the drug-loaded aqueous solution is dissolved with (3,3'-p-boronic acid phenyl methyl ammonium bromide)-methyl dipropylamine at a proportion of 1.5% W / W, mixed uniformly, and then polyvinyl alcohol solution is added at a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 4.5% W / W, mixed and stirred rapidly for 2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0075] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment. Specific embodiment four:

[0077] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel comprises the following steps:

[0078] S1. A certain mass of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, and then a certain volume of dimethylformamide solution is added, and the mixture is stirred under oil bath conditions for 3 h to obtain a mixture, which is used as needed;

[0079] Further, in step S1, the mass of N'N-bis(3-aminopropyl)methylamine is 0.11 g, the mass of 4-(bromomethyl)phenylboronic acid is 0.333 g, the volume of the dimethylformamide solution is 15 ml, and the concentration of the dimethylformamide solution is 98 wt%;

[0080] Further, in step S1, the oil bath temperature is 30°C, and the stirring speed is 400 rpm;

[0081] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1 in a volume ratio, and then placed in a-20°C refrigerator for 12 h to precipitate white solids, which are used as needed;

[0082] Further, in step S2, the volume ratio of the mixture to tetrahydrofuran is 15:40, the temperature of the ice tetrahydrofuran is-20°C, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1;

[0083] S3. The white solid obtained in step S2 was added to a certain volume of icy tetrahydrofuran, centrifuged and purified three times, and then freeze-dried in vacuo to obtain (3,3'-para-boronic acid benzyl ammonium bromide) - methyldipropylamine for use;

[0084] Furthermore, the centrifugal separation and purification treatment in step S3 is carried out under the conditions of centrifugation at 4000 rpm for 6 min at 4° C., and the vacuum freeze-drying treatment time is 12 h;

[0085] S4 weighing a certain mass of the loaded drug was added to ultrapure water to obtain a loaded drug aqueous solution for use;

[0086] Furthermore, in step S4, the loaded drug is glutamine, and the material-liquid ratio of the loaded drug to ultrapure water is 0.22 g:10 ml;

[0087] S5. Add (3,3'-p-boronic acid benzyl ammonium bromide)-methyldipropylamine obtained in step S3 to the drug-loaded aqueous solution obtained in step S4 according to the mass ratio. After mixing evenly, add a certain volume of polyvinyl alcohol solution, mix and rapidly stir for 1-2 minutes to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0088] Furthermore, in step S5, the drug-loaded aqueous solution is dissolved in (3,3'-para-boronic acid benzyl ammonium bromide)-methyldipropylamine at a ratio of 1.5% W / W, mixed evenly, and then a polyvinyl alcohol solution is added at a volume ratio of 1:2. The concentration of the polyvinyl alcohol solution is 4% W / W. The mixture is mixed and rapidly stirred for 1 minute to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0089] The intelligent dual-responsive drug-loaded myocardial injection hydrogel prepared by the preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel described in this embodiment is a dual-sensitive drug-releasing hydrogel carrier.

[0090] The hydrogel compositions of Examples 1 to 4 were tested for their mass remaining at pH 5.0 for five consecutive days. The test results were as follows: Figure 6 As shown, from Figure 6 Statistical results showed that the four different embodiments of the hydrogel had good pH sensitivity, and there was no statistical difference between the groups.

[0091] The hydrogels of Examples 1 to 4 were tested under 0.1 mM H2O2 conditions for five consecutive days. The test results are as follows: Figure 7 As shown, from Figure 7 It can be demonstrated that the four different embodiments of the hydrogel have good ROS sensitivity, and there is no statistical difference between the groups. DETAILED DESCRIPTION FIVE

[0093] A preparation method of an intelligent dual-response drug-loaded myocardial injection hydrogel, comprising the following steps:

[0094] S1. A certain mass of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenylboronic acid are weighed, and then a certain volume of dimethylformamide solution is added, and stirring is performed under oil bath conditions for 1 h to obtain a mixture, which is used as needed;

[0095] Further, in step S1, the mass of N'N-bis(3-aminopropyl)methylamine is 0.11 g, the mass of 4-(bromomethyl)phenylboronic acid is 0.333 g, the volume of the dimethylformamide solution is 15 ml, and the concentration of the dimethylformamide solution is 98 wt%;

[0096] Further, in step S1, the oil bath temperature is 35°C, and the stirring speed is 500 rpm;

[0097] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1 according to a volume ratio, and then placed in a-20°C refrigerator for 11 h to precipitate white solids, which are used as needed;

[0098] Further, in step S2, the volume ratio of the mixture to tetrahydrofuran is 18:45, the temperature of the ice tetrahydrofuran is-20°C, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1;

[0099] S3. The white solids obtained in step S2 are added to a certain volume of ice tetrahydrofuran, and then subjected to 2 times of centrifugal separation and purification treatment, and then subjected to vacuum freeze-drying treatment to obtain (3,3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine, which is used as needed;

[0100] Further, in step S3, the centrifugal separation and purification treatment is performed at 4°C at a speed of 4200 rpm for 5 min, and the vacuum freeze-drying treatment is performed for 11 h;

[0101] S4. A certain mass of a drug-loaded drug is weighed and added to ultrapure water to obtain a drug-loaded drug aqueous solution, which is used as needed;

[0102] Further, in step S4, the drug-loaded drug is glutamine, and the mass ratio of the drug-loaded drug to ultrapure water is 0.22 g:10 ml;

[0103] S5. The drug-loaded aqueous solution obtained in step S4 is added with (3,3'-p-boronic acid phenyl methyl ammonium bromide)-methyl dipropylamine in a mass ratio, mixed uniformly, and then a certain volume of polyvinyl alcohol solution is added, mixed and stirred rapidly for 2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0104] Further, in step S5, the drug-loaded aqueous solution is dissolved with (3,3'-p-boronic acid phenyl methyl ammonium bromide)-methyl dipropylamine at a proportion of 1.5% W / W, mixed uniformly, and then the polyvinyl alcohol solution is added at a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 3.5 W / W, mixed and stirred rapidly for 1 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0105] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment is used to prepare an intelligent dual-responsive drug-loaded myocardial injection hydrogel, and the intelligent dual-responsive drug-loaded myocardial injection hydrogel is a dual-sensitive drug release hydrogel carrier. Specific embodiment six:

[0107] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel comprises the following steps:

[0108] S1. A certain mass of N'N-bis(3-aminopropyl)methylamine and 4-(bromomethyl)phenyl boronic acid are weighed, and then a certain volume of dimethylformamide solution is added, and stirring is performed under oil bath conditions for 2.5 h to obtain a mixture, which is used as needed;

[0109] Further, in step S1, the mass of N'N-bis(3-aminopropyl)methylamine is 0.11 g, the mass of 4-(bromomethyl)phenyl boronic acid is 0.333 g, the volume of the dimethylformamide solution is 15 ml, and the concentration of the dimethylformamide solution is 98 wt%;

[0110] Further, in step S1, the oil bath temperature is 50°C, and the stirring speed is 300 rpm;

[0111] S2. A certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1 in a volume ratio, and then placed in a-20°C refrigerator for 12 h to precipitate white solids, which are used as needed;

[0112] Further, in step S2, the volume ratio of the mixture to tetrahydrofuran is 20:40, the temperature of the ice tetrahydrofuran is-20°C, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1;

[0113] S3. The white solid obtained in step S2 is added to a certain volume of ice in tetrahydrofuran, and then purified by centrifugal separation for 3 times, and then vacuum freeze-dried to obtain (3, 3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine, which is ready for use;

[0114] Further, the centrifugal separation purification treatment in step S3 is carried out at 4°C at a speed of 4500 rpm for 6 min, and the vacuum freeze-drying treatment is carried out for 14 h;

[0115] S4. A certain amount of drug-loaded is weighed and added to ultrapure water to obtain a drug-loaded aqueous solution, which is ready for use;

[0116] Further, the drug-loaded in step S4 is glutamine, and the ratio of drug-loaded to ultrapure water is 0.22 g:10 ml;

[0117] S5. The drug-loaded aqueous solution obtained in step S4 is added to (3, 3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine obtained in step S3 according to the mass ratio, mixed uniformly, and then a certain volume of polyvinyl alcohol solution is added, mixed and stirred rapidly for 1-2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0118] Further, in step S5, the drug-loaded aqueous solution is dissolved with (3, 3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine at a proportion of 1.5% W / W, mixed uniformly, and then a polyvinyl alcohol solution is added according to a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 4.5% W / W, and the mixture is mixed and stirred rapidly for 2 min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

[0119] The preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel according to the embodiment is used to prepare an intelligent dual-responsive drug-loaded myocardial injection hydrogel, and the intelligent dual-responsive drug-loaded myocardial injection hydrogel is a dual-sensitive drug release hydrogel carrier.

[0120] It should be noted that the terms "first" and "second" and the like relational terms are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between the entities or operations. Moreover, the terms "include", "contain" or any other variants thereof are intended to cover non-exclusive inclusion, so that the process, method, article or equipment including a series of elements not only includes those elements, but also includes other elements not explicitly listed or inherent to such process, method, article or equipment. Without more limitations, the element defined by the statement "including a" does not exclude the presence of another identical element in the process, method, article or equipment including the element.

[0121] Although the present application has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes can be made to the embodiments described and that equivalents can be substituted for elements thereof without departing from the scope of the present application. In particular, various features of the specific embodiments of the present application can be used in any combination, and the present application includes all such combinations, whether or not such combinations have been previously considered. Therefore, the present application is not limited to the specific embodiments disclosed herein, but includes all such embodiments falling within the scope of the claims.

Claims

1. A method for preparing an intelligent dual-responsive drug-loaded myocardial injection hydrogel, characterized in that, Comprising the following steps: S1. Weigh a certain mass of N'N-bis (3-aminopropyl) methylamine, 4- (bromomethyl) phenylboronic acid, and then add a certain volume of dimethylformamide solution, stir the reaction under oil bath conditions for 0.5h-4h to obtain a mixture, and wait for use; The mass of N'N-bis (3-aminopropyl) methylamine in step S1 is 0.11g, the mass of 4- (bromomethyl) phenylboronic acid is 0.333g, the volume of the dimethylformamide solution is 15ml, and the concentration of the dimethylformamide solution is 98wt%. The oil bath temperature in step S1 is 25℃-50℃, and the stirring speed is 200-500rpm; S2. According to the volume ratio, a certain volume of ice tetrahydrofuran is added to the mixture obtained in step S1, and then placed in a-20℃ refrigerator for 10h-12h to precipitate white solids, and then used; The volume ratio of the mixture and tetrahydrofuran in step S2 is 10-20:35-50, the temperature of the ice tetrahydrofuran is-20℃, and the volume ratio of the tetrahydrofuran added in step S3 to the tetrahydrofuran added in step S2 is 1:1; S3. The white solid obtained in step S2 is added to a certain volume of ice tetrahydrofuran, and then subjected to 2-3 times of centrifugal separation purification treatment, and then subjected to vacuum freeze-drying treatment to obtain (3, 3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine, and then used; The centrifugal separation purification treatment in step S3 is centrifuged at 4000-4500rpm for 4-6min at 4℃, and the vacuum freeze-drying treatment time is 10h-14h; S4. Weigh a certain mass of drug-loaded into ultrapure water to obtain a drug-loaded aqueous solution for use; The drug-loaded in step S4 is glutamine, and the feed liquid ratio of the drug-loaded and ultrapure water is 0.22g:10ml; S5. According to the mass ratio, (3, 3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine obtained in step S3 is added to the drug-loaded aqueous solution obtained in step S4, mixed uniformly, and then a certain volume of polyvinyl alcohol solution is added, mixed and stirred quickly for 1min-2min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel; In step S5, the drug-loaded aqueous solution is dissolved with (3, 3'-p-boronic acid benzyl ammonium bromide)-methyl dipropylamine at a ratio of 1.5%W / W, mixed uniformly, and then a polyvinyl alcohol solution is added according to a volume ratio of 1:2, the concentration of the polyvinyl alcohol solution is 3.5%W / W-4.5%W / W, and the mixture is mixed and stirred quickly for 1min-2min to obtain an intelligent dual-responsive drug-loaded myocardial injection hydrogel.

2. The intelligent dual-responsive drug-loaded myocardial injection hydrogel of claim 1, which is prepared by the preparation method of the intelligent dual-responsive drug-loaded myocardial injection hydrogel. The intelligent dual-responsive drug-loaded myocardial injection hydrogel is a dual-sensitive drug release hydrogel carrier.

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