In-situ temperature-sensitive gel drug-loaded preparation and preparation method thereof

Abstract

The invention relates to the technical field of medical biological materials, in particular to an in-situ temperature-sensitive gel drug-loaded preparation and a preparation method thereof, and the composition of the in-situ temperature-sensitive gel drug-loaded preparation comprises the following raw materials in percentage by mass: 10-50% of poloxamer 407; the composition further comprises any one of the following raw materials in percentage by mass: 1%-10% of poloxamer 188; or 1%-20% of hydroxypropyl methyl cellulose; or 0.1%-5% of polydopamine; or 0.03%-0.4% of a helical polyisocyanide polypeptide; or 0.1%-5% of polydopamine and 0.03%-0.4% of spiral polyisocyanide polypeptide. The in-situ temperature-sensitive gel drug-loaded preparation prepared by the invention has excellent temperature sensitivity, can keep flowing at low temperature, is convenient to apply, can be rapidly cured into gel at body temperature, realizes long-acting structural support and precise drug delivery, and remarkably improves the wound repair effect.

Description

Technical Field

[0001] The invention relates to the technical field of medical biomaterials, in particular to an in-situ temperature-sensitive gel drug-carrying preparation and a preparation method thereof. Background Art

[0002] In situ thermosensitive gel, as an advanced polymer material, has shown broad application prospects in the biomedical field due to its unique intelligent response characteristics. Its core advantage is that it can achieve sol-gel phase transition according to changes in ambient temperature. At room temperature or low temperature, this gel remains in liquid state, which is easy to apply by injection or smearing. When the ambient temperature rises to close to human body temperature, it will quickly transform into a stable gel state to form a three-dimensional structure. This feature makes in situ thermosensitive gel have great potential in many fields such as drug delivery systems, wound management and tissue engineering.

[0003] As a typical thermosensitive block copolymer, Poloxamer 407 is composed of hydrophilic ethylene oxide (PEO) and hydrophobic propylene oxide (PPO). It has excellent temperature sensitivity and can quickly gel at body temperature. It is widely used in the development of thermosensitive gels. However, despite its excellent performance in temperature response, Poloxamer 407 still faces a series of challenges in practical applications.

[0004] First, the lack of mechanical properties limits its scope of application. The gel formed by poloxamer 407 contains a high water content, resulting in relatively low mechanical strength in the gel state, which makes it difficult to provide sufficient structural support. This defect is particularly evident in applications where large wounds are required or structural stability needs to be maintained for a long time. Secondly, poor tissue adhesion is also a problem that needs to be solved urgently. Although high water content helps to create a moist healing environment, poloxamer 407 has weak adhesion to the tissue surface and is easily detached from the wound surface under the action of body fluids or external forces. This will not only affect the continuous delivery of the drug, but may also weaken the healing effect. In addition, the limitations of biocompatibility cannot be ignored. Although the thermosensitive gel using poloxamer 407 alone has certain biocompatibility, it is quite different from the mechanical and chemical properties of biological tissues and cannot effectively imitate the stress stiffening properties of biological tissues. This limits its application effect in complex biological environments to a certain extent.

[0005] In summary, the existing in situ thermosensitive gel based on poloxamer 407 still has significant room for improvement in terms of mechanical properties and bioadhesion. In order to fully realize its potential in the biomedical field, further exploration and optimization of material formulation and preparation process are needed to overcome the above challenges. Summary of the invention

[0006] The purpose of the present invention is to provide an in-situ thermosensitive gel drug-loaded preparation and a preparation method thereof in order to solve at least one of the above technical problems.

[0007] The present invention achieves the above-mentioned purpose through the following technical solutions:

[0008] An in-situ thermosensitive gel drug-carrying preparation, the composition of which comprises the following raw materials in percentage by mass: 10%-50% of poloxamer 407;

[0009] It also includes any of the following raw materials in a mass percentage ratio:

[0010] 1%-10% Poloxamer 188;

[0011] Alternatively, 1%-20% hydroxypropyl methylcellulose;

[0012] Alternatively, 0.1%-5% polydopamine;

[0013] Alternatively, 0.03%-0.4% helical polyisocyanide polypeptide;

[0014] Alternatively, 0.1%-5% polydopamine and 0.03%-0.4% helical polyisocyanide polypeptide.

[0015] Furthermore, the gelling temperature of the in situ thermosensitive gel drug-loaded preparation is 25°C-38°C.

[0016] Furthermore, in the poloxamer 407, EO:PO=7:3.

[0017] Furthermore, the mass percentage of Poloxamer 407 in the composition is 10%-30%.

[0018] Furthermore, the mass percentage of polydopamine in the composition is 1%-5%.

[0019] Furthermore, the helical polyisocyanide polypeptide chain length in the composition is 0.5K-5K.

[0020] Furthermore, dopamine hydrochloride was dissolved in 1x PBS at pH=8, and self-polymerized into polydopamine under alkaline conditions, which was prepared and used immediately.

[0021] A method for preparing an in-situ thermosensitive gel drug-loaded preparation, used to prepare any of the above in-situ thermosensitive gel drug-loaded preparations, the method comprising the following steps:

[0022] Dissolve Poloxamer 407 in ddH2O at low temperature to prepare a stock solution, and let it stand for a day and night to dissolve for later use;

[0023] When the raw materials of the composition include poloxamer 188, the poloxamer 188 solution is dissolved in ddH2O at room temperature to prepare a stock solution for standby use;

[0024] When the raw materials of the composition include hydroxypropyl methylcellulose, the hydroxypropyl methylcellulose is dissolved in ddH2O at room temperature to prepare a stock solution, and the solution is allowed to stand for one day and night to dissolve for later use;

[0025] When the raw materials of the composition include polydopamine, dopamine hydrochloride is dissolved to self-polymerize into polydopamine under alkaline conditions, and the polydopamine is prepared and used immediately;

[0026] When the raw materials of the composition include a helical polyisocyanate polypeptide, the helical polyisocyanate polypeptide is dissolved in 1x PBS to prepare a stock solution for later use;

[0027] The raw materials required for the composition are uniformly mixed to obtain an in-situ thermosensitive gel drug-carrying preparation.

[0028] Further, hydroxypropyl methylcellulose is dissolved in ddH2O at 30-50°C.

[0029] The beneficial effects of the present invention are:

[0030] The present invention provides an in-situ thermosensitive gel drug-carrying preparation and a preparation method thereof, wherein the drug-carrying preparation combines the temperature responsiveness of poloxamer, the biocompatibility and mechanical biomimetic properties of helical polyisocyanurate polypeptide and polydopamine, and the thickening and stability of hydroxypropyl methylcellulose, thereby creating a high-performance in-situ thermosensitive gel drug-carrying preparation. The in-situ thermosensitive gel drug-carrying preparation not only has excellent temperature sensitivity and can quickly form a stable gel structure at body temperature, but also significantly improves mechanical biomimetic properties and tissue adhesion, providing an ideal physiological microenvironment for medical applications such as drug delivery, injection therapy, and wound repair, effectively prolonging the drug action time, enhancing the therapeutic effect, and showing broad prospects for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a flow chart of a method for preparing an in-situ thermosensitive gel drug-loaded preparation according to one embodiment of the present invention;

[0032] Figure 2 The test results of measuring the complex viscosity and storage modulus of poloxamer 407 hydrogels with different ratios by TA-DHR2 rheometer according to one embodiment of the present invention;

[0033] Figure 3 The test results of composite viscosity and storage modulus of poloxamer 407 and poloxamer 188 hydrogels with different ratios measured by TA-DHR2 rheometer according to one embodiment of the present invention;

[0034] Figure 4 The test results of composite viscosity and storage modulus of poloxamer 407 and hydroxypropyl methylcellulose hydrogels in different ratios measured by TA-DHR2 rheometer according to one embodiment of the present invention;

[0035] Figure 5 This is a test result 1 of measuring the composite viscosity and storage modulus of poloxamer 407 and polydopamine hydrogels in different ratios by a TA-DHR2 rheometer according to an embodiment of the present invention;

[0036] Figure 6 The second test result of measuring the composite viscosity and storage modulus of poloxamer 407 and polydopamine hydrogels in different ratios by TA-DHR2 rheometer according to one embodiment of the present invention;

[0037] Figure 7 The test results of composite viscosity and storage modulus of poloxamer 407 and helical polyisocyanurate polypeptide hydrogels with different ratios measured by TA-DHR2 rheometer according to one embodiment of the present invention;

[0038] Figure 8 The test results of the composite viscosity and storage modulus of a mixed hydrogel of poloxamer 407 and helical polyisocyanurate polypeptides of different chain lengths measured by a TA-DHR2 rheometer according to one embodiment of the present invention;

[0039] Fig. 9 The particle size changes of the helical polyisocyanate polypeptide, polydopamine, and the assembly of the helical polyisocyanate polypeptide and polydopamine according to one embodiment of the present invention;

[0040] Fig.10 This is a SEM image of an in-situ thermosensitive gel drug-carrying preparation according to one embodiment of the present invention;

[0041] Fig.11 This is a schematic diagram of an in-situ thermosensitive gel drug-carrying preparation according to one embodiment of the present invention changing from a liquid state to a gel state on a hand;

[0042] Fig.12 This is a schematic diagram of the state change of an in-situ thermosensitive gel drug-loaded preparation before and after gelation according to one embodiment of the present invention. DETAILED DESCRIPTION

[0043] The content of the present invention will now be discussed with reference to exemplary embodiments. It should be understood that the embodiments discussed are only to enable those skilled in the art to better understand and thus implement the content of the present invention, rather than implying any limitation on the scope of the present invention.

[0044] As used herein, the term “including” and variations thereof are to be interpreted as open-ended terms meaning “including, but not limited to.” The term “based on” is to be interpreted as “based, at least in part, on.” The terms “one embodiment” and “an embodiment” are to be interpreted as “at least one embodiment.”

[0045] Embodiment 1

[0046] In situ thermosensitive gel technology is an intelligent drug delivery system. Its key characteristic is temperature responsiveness, that is, the material undergoes a physical or chemical structural change when the ambient temperature changes, forming a gel combination that adapts to the microenvironment of the lesion, providing a physiological environment similar to that of the tissue, and prolonging the duration of the drug's action. Although the commonly used poloxamer 407 has a good temperature response, it lacks the mechanical biomimetic properties of biological tissues. The present invention is based on poloxamer, helical polyisocyanurate polypeptides and polydopamine, hydroxypropyl methylcellulose to prepare an in situ thermosensitive gel drug-carrying preparation with high biocompatibility, excellent mechanical properties, and strong adhesion, which can be used in the fields of drug loading, injection, wound repair, etc.

[0047] According to one embodiment of the present invention, an in situ thermosensitive gel drug-carrying preparation comprises the following raw materials in percentage by mass: 10%-50% of poloxamer 407;

[0048] It also includes any of the following raw materials in a mass percentage ratio:

[0049] 1%-10% Poloxamer 188;

[0050] Alternatively, 1%-20% hydroxypropyl methylcellulose;

[0051] Alternatively, 0.1%-5% polydopamine;

[0052] Alternatively, 0.03%-0.4% helical polyisocyanide polypeptide;

[0053] Alternatively, 0.1%-5% polydopamine and 0.03%-0.4% helical polyisocyanide polypeptide.

[0054] Preferably, the gelling temperature of the in situ thermosensitive gel drug-loaded preparation is 25°C-38°C.

[0055] Preferably, in poloxamer 407, EO:PO=7:3.

[0056] In this embodiment, an in situ thermosensitive gel drug-loaded preparation is provided for wound repair. The raw materials for preparing the in situ thermosensitive gel drug-loaded preparation include poloxamer 407, poloxamer 188, hydroxypropyl methylcellulose, polydopamine, and helical polyisocyanurate polypeptide; the prepared in situ thermosensitive gel drug-loaded preparation has various forms according to the different raw materials used, and the composition includes the following raw materials in mass percentage:

[0057] In-situ thermosensitive gel drug-carrying preparation 1, the raw materials include 10%-50% by weight of poloxamer 407 and 1%-10% by weight of poloxamer 188.

[0058] In-situ thermosensitive gel drug-carrying preparation 2, the raw materials include 10%-50% by mass of poloxamer 407 and 1%-20% by mass of hydroxypropyl methylcellulose. Among them, hydroxypropyl methylcellulose has low viscosity. Poloxamer 407 and hydroxypropyl methylcellulose are combined to form a biomimetic thermosensitive gel preparation.

[0059] In-situ thermosensitive gel drug-carrying preparation 3, the raw materials include 10%-50% by weight of poloxamer 407 and 0.1%-5% by weight of polydopamine.

[0060] In situ thermosensitive gel drug-carrying preparation 4, the raw materials include 10%-50% by mass of poloxamer 407 and 0.03%-0.4% of helical polyisocyanate polypeptide. Helical polyisocyanate polypeptide has a stable helical structure. As a new type of biomimetic polymer material, helical polyisocyanate polypeptide exhibits excellent biocompatibility, thermosensitivity and stress stiffening properties similar to biopolymers, significantly enhancing its mechanical properties and tissue adhesion. The material imitates the mechanical behavior of natural biological tissues, especially in stress stiffening and structural elasticity. Its unique helical structure can simulate the strain response of biological tissues when subjected to stress and provide strong mechanical support. Helical polyisocyanate polypeptide effectively absorbs and releases energy when encountering external pressure or mechanical stress through the extension and compression of the helical structure. This process is similar to the stiffening and rebound mechanism of natural tissues when subjected to stress. This feature is particularly important in applications such as wound repair, because it can provide mechanical support similar to that of natural tissues and effectively resist structural damage that may be caused by movement or pressure in the body. In addition, helical polyisocyanurate peptides also have the ability to achieve sol-gel transition with temperature changes, similar to poloxamer, and can work synergistically with other temperature-sensitive materials to further enhance the physical and chemical stability of the gel.

[0061] In situ thermosensitive gel drug-loaded preparation 5, the raw materials include 10%-50% by mass of poloxamer 407, 0.1%-5% of polydopamine and 0.03%-0.4% of helical polyisocyanate polypeptide. Under alkaline conditions, dopamine hydrochloride self-polymerizes and self-assembles with the helical structure of the helical polyisocyanate polypeptide to form a complex. This process significantly enhances the adhesion of the thermosensitive gel to the wound tissue. By combining the helical polyisocyanate polypeptide and the polydopamine assembly, not only the structural stability and mechanical biomimetic properties of the gel are greatly improved, but also a more bionic tissue repair environment is created, overcoming the deficiencies of existing thermosensitive gels in mechanical properties and adhesion.

[0062] The in situ thermosensitive gel drug-loaded preparation prepared by the present invention has excellent temperature sensitivity, can remain fluid at low temperatures for easy application, and quickly solidifies into a gel at body temperature, thereby achieving long-term structural support and precise drug delivery, and significantly improving the wound repair effect.

[0063] According to one embodiment of the present invention, the mass percentage of Poloxamer 407 in the composition is 10%-30%.

[0064] Preferably, the mass percentage of polydopamine in the composition is 1%-5%.

[0065] Preferably, the helical polyisocyanide polypeptide chain length in the composition is 0.5K-5K.

[0066] Preferably, dopamine hydrochloride is dissolved in 1x PBS at pH=8, so that it self-polymerizes into polydopamine under alkaline conditions and is ready for use.

[0067] In this embodiment, the in-situ thermosensitive gel drug-carrying preparation is a thermosensitive hydrogel based on poloxamer 407, which is specially designed for wound repair. Its characteristic is that it can be mixed with poloxamer 188, hydroxypropyl methylcellulose, polydopamine self-polymerized under alkaline conditions, and helical polyisocyanurate polypeptides with a chain length of 0.5K-5K to prepare a variety of wound dressings. These dressings have shown wide application potential in the field of wound repair by virtue of their unique properties.

[0068] The in-situ thermosensitive gel drug-carrying preparation provided by the present invention is based on poloxamer 407 and significantly improves the effect and application potential of wound repair by adding multiple reinforcing ingredients.

[0069] Embodiment 2

[0070] Figure 1 FIG. 1 is a flow chart of a method for preparing an in situ thermosensitive gel drug-loaded preparation according to an embodiment of the present invention. Figure 1 As shown, according to one embodiment of the present invention, a method for preparing an in situ thermosensitive gel drug-loaded preparation is used for any in situ thermosensitive gel drug-loaded preparation of the present invention, and the method comprises the following steps:

[0071] Step S1, dissolving poloxamer 407 in ddH2O at low temperature to prepare a stock solution, and letting it stand for one day and night to dissolve for later use;

[0072] Step S2, when the raw materials of the composition include poloxamer 188, dissolving the poloxamer 188 solution in ddH2O at room temperature to prepare a stock solution for standby use;

[0073] Step S3, when the raw materials of the composition include hydroxypropyl methylcellulose, dissolving the hydroxypropyl methylcellulose in ddH2O at room temperature to prepare a stock solution, and standing it for one day and night to dissolve for use;

[0074] Step S4, when the raw materials of the composition include polydopamine, dissolving dopamine hydrochloride to allow it to self-polymerize under alkaline conditions to form polydopamine, which is prepared and used immediately;

[0075] Step S5, when the raw materials of the composition include a helical polyisocyanate polypeptide, dissolving the helical polyisocyanate polypeptide in 1xPBS to prepare a stock solution for standby use;

[0076] Step S6, uniformly mixing the raw materials required for the composition to obtain an in-situ thermosensitive gel drug-loaded preparation.

[0077] Preferably, in step S3, hydroxypropyl methylcellulose is dissolved in ddH2O at 30-50°C.

[0078] In this embodiment, a method for preparing an in situ thermosensitive gel drug-loaded preparation for wound repair is provided, based on poloxamer 407, comprising the following steps:

[0079] Step S1, dissolve poloxamer 407 with a mass fraction of 10%-50% in pre-cooled ddH2O under low temperature conditions to prepare a storage solution, and let it stand for a day and night to dissolve for use. Step S2: dissolve poloxamer 188 solution with a mass fraction of 1%-10% in pre-cooled ddH2O under low temperature conditions to prepare a storage solution for use. Step S3: dissolve hydroxypropyl methylcellulose with a mass fraction of 1%-20% in ddH2O at room temperature to prepare a storage solution, and let it stand for a day and night to dissolve for use. Hydroxypropyl methylcellulose has low viscosity and needs to be stirred and dispersed with hot water first, and then cold water is added to stir and cool to dissolve. Therefore, hydroxypropyl methylcellulose is dissolved in ddH2O at 30-50°C to prevent caking and wrapping. Stir quickly and let it stand for a day and night to eliminate bubbles. Step S4: dissolve dopamine hydrochloride in 1x PBS with pH=8, so that it self-polymerizes into polydopamine under alkaline conditions, and prepare it for immediate use. Dopamine hydrochloride self-polymerizes into polydopamine under alkaline conditions, and in order to maintain the stability of alkaline conditions, 1x PBS is used in this embodiment. Step S5: dissolve the helical polyisocyanate polypeptide in 1x PBS to prepare a storage solution for standby use. Step S6: The storage solutions prepared in steps S2-S5 are respectively mixed with the storage solutions prepared in step S1 in corresponding proportions to obtain a thermosensitive hydrogel for wound repair based on poloxamer 407. Specifically, the in situ thermosensitive gel drug-loaded preparation 1, the raw materials include 10%-50% by mass of poloxamer 407 and 1%-10% by mass of poloxamer 188. In situ thermosensitive gel drug-carrying preparation 2, the raw materials include 10%-50% by mass of poloxamer 407 and 1%-20% by mass of hydroxypropyl methylcellulose. In situ thermosensitive gel drug-carrying preparation 3, the raw materials include 10%-50% by mass of poloxamer 407 and 0.1%-5% by mass of polydopamine. In situ thermosensitive gel drug-carrying preparation 4, the raw materials include 10%-50% by mass of poloxamer 407 and 0.03%-0.4% of helical polyisocyanate polypeptide. In situ thermosensitive gel drug-carrying preparation 5, the raw materials include 10%-50% by mass of poloxamer 407, 0.1%-5% of polydopamine and 0.03%-0.4% of helical polyisocyanate polypeptide.

[0080] The preparation method of the in-situ thermosensitive gel drug-loaded preparation in this embodiment is based on the principle that the poloxamer 407-based polymer is composed of hydrophilic ethylene oxide (EO) and hydrophobic propylene oxide (PO) blocks, presenting a triblock structure (PEO-PPO-PEO), and undergoes a sol-gel transition as the temperature rises. Poloxamer 188 is a nonionic block linear copolymer with rheological activity, which has a certain cell protection function and can reduce the gel temperature of the system. The hydroxyl group on hydroxypropyl methylcellulose is a hydrophilic group, and the methoxy group is a hydrophobic group. As the temperature rises, the solution absorbs heat and gradually removes the water molecules bound to the cellulose molecular chain, exposing the hydrophobic group, and the molecules form a bond due to the hydrophobic effect, so that the solution becomes a turbid liquid state and then further transforms into a network gel structure. Dopamine hydrochloride self-polymerizes into polydopamine (PDA) under alkaline conditions, which has excellent biocompatibility, antioxidant activity and wet adhesion. Polyisocyanurate polypeptide hydrogels are connected to oligoethylene glycol on their main chains. The hydrophobic interactions between the oligoethylene glycol tails enable rapid and reversible transition from liquid to gel. As a new type of synthetic polymer material with good biocompatibility, thermosensitivity, and stress stiffening properties similar to those of biopolymers, it overcomes the shortcomings of traditional hydrogels, such as poor mechanical properties and weak tissue adhesion.

[0081] The present invention not only significantly enhances the mechanical properties of the thermosensitive hydrogel, but also flexibly adjusts the gelling temperature by precisely regulating the ratio of poloxamer 407 to poloxamer 188, hydroxypropyl methylcellulose, polydopamine (formed by alkaline self-polymerization of dopamine hydrochloride), and helical polyisocyanurate polypeptide, thereby preparing a high-performance dressing suitable for wound repair. These dressings combine excellent biocompatibility, thermosensitive responsiveness, and enhanced tissue adhesion, effectively overcoming the limitations of traditional hydrogels in mechanical properties and adhesion, and providing a more reliable and efficient solution for wound repair.

[0082] Embodiment 3

[0083] Example 1 Poloxamer 407 hydrogel

[0084] According to one embodiment of the present invention, an in-situ thermosensitive gel drug-loaded preparation includes: 20%-25% by mass of poloxamer 407. Specifically, the preparation method thereof includes the following steps:

[0085] Step S1: dissolving poloxamer 407 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0086] Step S2: dilute the poloxamer 407 stock solution into hydrogels with a mass fraction of 20%-25%.

[0087] The prepared in-situ thermosensitive gel drug-loaded preparation is tested, and the testing method comprises the following steps:

[0088] The gel samples with different mass fractions were measured using a TA-DHR2 rheometer. The rheometer parameters were oscillation mode, starting temperature of 20°C, ending temperature of 50°C, temperature change time of 5°C / min, strain of 1%, and frequency of 1Hz.

[0089] Figure 2 The test results of the composite viscosity and storage modulus of poloxamer 407 hydrogels with different ratios measured by TA-DHR2 rheometer according to one embodiment of the present invention; wherein, Figure 2 (a) shows the change of the complex viscosity of 21%-25% Poloxamer 407 with temperature. Figure 2 (b) shows the change of storage modulus of 21%-25% poloxamer 407 with temperature. Figure 2 As shown, the greater the mass fraction of Poloxamer 407, the lower the gel temperature and the higher the complex viscosity and storage modulus.

[0090] Example 2: Poloxamer 407 and Poloxamer 188 mixed hydrogel

[0091] According to one embodiment of the present invention, an in situ thermosensitive gel drug-loaded preparation comprises: 21% by mass of poloxamer 407 and 1%-10% by mass of poloxamer 188. Specifically, the preparation method thereof comprises the following steps:

[0092] Step S1: dissolving poloxamer 407 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0093] Step S2: dissolving poloxamer 188 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0094] Step S3: Poloxamer 188 is mixed with poloxamer 407 in a certain proportion in sequence, so that the mass fraction of poloxamer 188 in the sample is 1%-10%, and the mass fraction of poloxamer 407 in the sample is 21%.

[0095] The prepared in-situ thermosensitive gel drug-loaded preparation is tested, and the testing method comprises the following steps:

[0096] The different samples were measured using a TA-DHR2 rheometer. The rheometer parameters were oscillation mode, starting temperature of 20°C, ending temperature of 50°C, temperature change time of 5°C / min, strain of 1%, and frequency of 1 Hz.

[0097] Figure 3The composite viscosity and storage modulus of poloxamer 407 and poloxamer 188 hydrogels in different ratios were measured by a TA-DHR2 rheometer according to an embodiment of the present invention; wherein, Figure 3 (a) shows the change of the composite viscosity of the hydrogel mixed with poloxamer 407 and poloxamer 188 in different ratios with temperature. Figure 3 (b) shows the change of storage modulus of hydrogels mixed with poloxamer 407 and poloxamer 188 in different ratios with temperature. Figure 3 As shown in the figure, when poloxamer 188 is mixed with poloxamer 407, the gelation temperature is reduced. The larger the mass fraction of poloxamer 188, the lower the gelation temperature, and the higher the complex viscosity and storage modulus.

[0098] Example 3: Poloxamer 407 and Hydroxypropyl Methylcellulose Mixed Hydrogel

[0099] According to one embodiment of the present invention, an in situ thermosensitive gel drug-carrying preparation comprises: 24% by mass of poloxamer 407 and 1% to 20% by mass of hydroxypropyl methylcellulose. Specifically, the preparation method thereof comprises the following steps:

[0100] Step S1: dissolving poloxamer 407 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0101] Step S2: dissolving hydroxypropyl methylcellulose in ddH2O at room temperature to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for later use;

[0102] Step S3: Poloxamer 407 is sequentially mixed with hydroxypropyl methylcellulose in a certain proportion, so that the mass fraction of hydroxypropyl methylcellulose in the sample is 1%-20%, and the mass fraction of poloxamer 407 in the sample is 24%.

[0103] The prepared in-situ thermosensitive gel drug-loaded preparation is tested, and the testing method comprises the following steps:

[0104] The different samples were measured using a TA-DHR2 rheometer. The rheometer parameters were oscillation mode, starting temperature of 20°C, ending temperature of 50°C, temperature change time of 5°C / min, strain of 1%, and frequency of 1 Hz.

[0105] Figure 4 The composite viscosity and storage modulus of poloxamer 407 and hydroxypropyl methylcellulose hydrogels in different ratios were measured by a TA-DHR2 rheometer according to an embodiment of the present invention; wherein, Figure 4 (a) shows the change of the composite viscosity of the hydrogel mixed with different proportions of poloxamer 407 and hydroxypropyl methylcellulose with temperature. Figure 4 (b) shows the change of storage modulus of hydrogels mixed with different proportions of poloxamer 407 and hydroxypropyl methylcellulose with temperature. Figure 4 As shown, hydroxypropyl methylcellulose reduces the gel temperature, enhances the composite viscosity and storage modulus, the greater the mass fraction of hydroxypropyl methylcellulose, the lower the gel temperature, HPMC is a thickening agent, the greater the mass fraction, the stronger the hydrophobicity, and the lower the gel temperature.

[0106] Example 4: Poloxamer 407 and polydopamine mixed hydrogel

[0107] According to one embodiment of the present invention, an in situ thermosensitive gel drug-loaded preparation comprises: 25% by mass of poloxamer 407 and 0.1% to 5% by mass of polydopamine. Specifically, the preparation method thereof comprises the following steps:

[0108] Step S1: dissolving poloxamer 407 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0109] Step S2: dissolving dopamine hydrochloride in a NaOH solution with a pH of 8, so that it self-polymerizes under alkaline conditions to form polydopamine having biocompatibility, antioxidant activity and wet adhesion, which is used immediately after preparation;

[0110] Step S3: polydopamine is sequentially mixed with poloxamer 407 in a certain proportion, so that the mass fraction of polydopamine in the sample is 0.1%-5%, and the mass fraction of poloxamer 407 in the sample is 25%.

[0111] The prepared in-situ thermosensitive gel drug-loaded preparation is tested, and the testing method comprises the following steps:

[0112] The different samples were measured using a TA-DHR2 rheometer. The rheometer parameters were oscillation mode, starting temperature of 20°C, ending temperature of 50°C, temperature change time of 5°C / min, strain of 1%, and frequency of 1 Hz.

[0113] Figure 5 This is a test result 1 of measuring the composite viscosity and storage modulus of poloxamer 407 and polydopamine hydrogels in different ratios by TA-DHR2 rheometer according to an embodiment of the present invention; wherein, Figure 5 (a) shows the change of the composite viscosity of the hydrogel mixed with poloxamer 407 and 1%-5% polydopamine with temperature. Figure 5 (b) The storage modulus of the hydrogel mixed with poloxamer 407 and 1%-5% polydopamine changes with temperature. Figure 6The second test result of measuring the composite viscosity and storage modulus of poloxamer 407 and polydopamine hydrogels in different ratios by TA-DHR2 rheometer according to an embodiment of the present invention; wherein, Figure 6 (a) shows the change of the composite viscosity of the hydrogel mixed with poloxamer 407 and 0.125%-0.5% polydopamine with temperature. Figure 6 (b) shows the change of storage modulus of hydrogel mixed with poloxamer 407 and 0.125%-0.5% polydopamine with temperature. Figure 5-6 As shown, when the mass fraction of polydopamine is above 1%, the larger the mass fraction of polydopamine, the higher the gel temperature, and the lower the complex viscosity and storage modulus; when the mass fraction of polydopamine is below 1%, the smaller the mass fraction of polydopamine, the higher the gel temperature, and the lower the complex viscosity and storage modulus.

[0114] Example 5: Poloxamer 407 and helical polyisocyanurate polypeptide mixed hydrogel

[0115] According to one embodiment of the present invention, an in situ thermosensitive gel drug-carrying preparation comprises: poloxamer 407 with a mass fraction of 25% and a helical polyisocyanide polypeptide with a concentration of 0.35 mg / ml. Specifically, the preparation method thereof comprises the following steps:

[0116] Step S1: dissolving poloxamer 407 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0117] Step S2: dissolving the pre-synthesized helical polyisocyanide polypeptide in 1X PBS to a 4 mg / ml stock solution;

[0118] Step S3: The helical polyisocyanate polypeptide is sequentially mixed with poloxamer 407 in a certain ratio, so that the final concentration of the helical polyisocyanate polypeptide in the sample is 0.35 mg / ml, and the mass fraction of poloxamer 407 in the sample is 25%.

[0119] The prepared in-situ thermosensitive gel drug-loaded preparation is tested, and the testing method comprises the following steps:

[0120] The different samples were measured using a TA-DHR2 rheometer. The rheometer parameters were oscillation mode, starting temperature of 20°C, ending temperature of 50°C, temperature change time of 5°C / min, strain of 1%, and frequency of 1 Hz.

[0121] Figure 7 The composite viscosity and storage modulus of poloxamer 407 and helical polyisocyanide polypeptide hydrogels in different ratios were measured by TA-DHR2 rheometer according to one embodiment of the present invention; wherein, Figure 7(a) shows the change of the composite viscosity of the hydrogel mixed with poloxamer 407 and helical polyisocyanurate polypeptide with temperature. Figure 7 (b) The storage modulus of the hydrogel mixed with poloxamer 407 and helical polyisocyanurate polypeptide changes with temperature.

[0122] Example 6: Poloxamer 407 and different chain length helical polyisocyanurate polypeptide mixed hydrogel

[0123] According to one embodiment of the present invention, an in situ thermosensitive gel drug-carrying preparation comprises: poloxamer 407 with a mass fraction of 20%, polydopamine with a concentration of 2%, and helical polyisocyanide polypeptide with a concentration of 4 mg / ml. Specifically, the preparation method thereof comprises the following steps:

[0124] Step S1: dissolving poloxamer 407 in pre-cooled ddH2O under low temperature conditions to prepare a stock solution, and letting it stand for one day and night until it is fully dissolved for use;

[0125] Step S2: dissolving the pre-synthesized helical polyisocyanide polypeptide in 1X PBS to a 4 mg / ml stock solution;

[0126] Step S3: polydopamine and helical polyisocyanate polypeptide are mixed in a certain proportion in sequence, so that the final concentration of polydopamine in the sample is 2%, the final concentration of helical polyisocyanate polypeptide in the sample is 4 mg / ml, and the mass fraction of poloxamer 407 in the sample is 20%.

[0127] The prepared in-situ thermosensitive gel drug-loaded preparation is tested, and the testing method comprises the following steps:

[0128] The different samples were measured using a TA-DHR2 rheometer. The rheometer parameters were oscillation mode, starting temperature of 20°C, ending temperature of 50°C, temperature change time of 5°C / min, strain of 1%, and frequency of 1 Hz.

[0129] Figure 8 The composite viscosity and storage modulus of a mixed hydrogel of poloxamer 407 and helical polyisocyanurate polypeptides of different chain lengths were measured by a TA-DHR2 rheometer according to one embodiment of the present invention; wherein, Figure 8 (a) shows the change of the composite viscosity of the hydrogel mixed with poloxamer 407 and helical polyisocyanurate polypeptides of different chain lengths with temperature. Figure 8 (b) The storage modulus of the hydrogel mixed with poloxamer 407 and helical polyisocyanurate polypeptides of different chain lengths changes with temperature. Fig. 9 The particle size changes of the helical polyisocyanate polypeptide, polydopamine, and the helical polyisocyanate polypeptide and polydopamine assembly according to one embodiment of the present invention are shown in FIG. Figure 8-9As shown, the chain length of the helical polyisocyanate polypeptide has an effect on the in situ thermosensitive gel drug-carrying preparation. Under the same conditions, the gel point temperature of the helical polyisocyanate polypeptide with a chain length of 0.5K is higher than the gel point temperature of the helical polyisocyanate polypeptide with a chain length of 3K, indicating that the longer the chain, the lower the gel point. Therefore, in practical applications, we use PIC with a chain length of 0.5K-5K. Since the mechanical strength in the gel state is low after adding PIC, sufficient structural support is provided, and at the same time, the longer the PIC chain, the lower the gel point. The mass percentage of PIC used in the present invention is 0.03%-0.4%, and the chain length is 0.5K-5K.

[0130] Fig.10 This is a SEM image of an in-situ thermosensitive gel drug-carrying preparation according to one embodiment of the present invention; Fig.11 This is a schematic diagram of an in-situ thermosensitive gel drug-carrying preparation according to one embodiment of the present invention changing from a liquid state to a gel state on a hand; Fig.12 Schematic diagram of the state change of the in-situ thermosensitive gel drug-carrying preparation before and after gelation according to one embodiment of the present invention. Figure 10-12 The in-situ thermosensitive gel drug-carrying preparation provided by the present invention has broad application prospects and important research value in the field of biomedicine, specifically including:

[0131] Excellent biocompatibility and thermosensitive properties: The in situ thermosensitive gel drug-loaded preparation of the present invention not only has excellent biocompatibility, but also exhibits stress stiffening properties comparable to those of biopolymers, significantly enhancing its mechanical properties and adhesion to tissues, providing a solid foundation for biomedical applications.

[0132] Efficient wound protection barrier: As a protective barrier, the in situ thermosensitive gel drug-loaded preparation of the present invention can effectively cover and protect open wounds, reduce external contamination and infection risks, and create a safer and cleaner environment for wound healing.

[0133] Sustained drug release and improved utilization rate: The temperature sensitivity of the in situ thermosensitive gel drug-loaded preparation of the present invention enables the drug to be slowly released in vivo, prolonging the administration time, thereby significantly improving the utilization rate and therapeutic effect of the drug, and bringing more lasting and stable therapeutic effects to patients.

[0134] Simple preparation and high cost-effectiveness: The in situ thermosensitive gel drug-loaded preparation of the present invention is prepared by a relatively simple and low-cost method, which not only reduces the production cost but also improves the production efficiency, making the preparation have a wider application prospect in the medical field.

[0135] Excellent antibacterial properties: The in situ thermosensitive gel drug-loaded preparation of the present invention also exhibits excellent antibacterial properties, which can further reduce the risk of wound infection and provide patients with more comprehensive and effective treatment protection.

[0136] The above description is only a preferred embodiment of the present application and an explanation of the technical principles used. Those skilled in the art should understand that the scope of the invention involved in the present application is not limited to the technical solution formed by a specific combination of the above technical features, but should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the inventive concept. For example, the above features are replaced with (but not limited to) technical features with similar functions disclosed in the present application.

[0137] It should be understood that the size of the serial numbers of each step in the content of the invention and the embodiments of the present invention does not absolutely mean the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present invention.

Claims

1. An in situ thermosensitive gel drug-loaded preparation, characterized in that: The composition includes the following raw materials in percentage by mass: 10%-50% of poloxamer 407; It also includes any of the following raw materials in a mass percentage ratio: 1%-10% Poloxamer 188; Alternatively, 1%-20% hydroxypropyl methylcellulose; Alternatively, 0.1%-5% polydopamine; Alternatively, 0.03%-0.4% helical polyisocyanide polypeptide; Alternatively, 0.1%-5% polydopamine and 0.03%-0.4% helical polyisocyanide polypeptide.

2. The in situ thermosensitive gel drug-loaded preparation according to claim 1, characterized in that: The gelling temperature of the in-situ thermosensitive gel drug-carrying preparation is 25°C-38°C.

3. The in situ thermosensitive gel drug-loaded preparation according to claim 1, characterized in that: In the poloxamer 407, EO:PO=7:

3.

4. The in situ thermosensitive gel drug-loaded preparation according to claim 1, characterized in that: The mass percentage of Poloxamer 407 in the composition is 10%-30%.

5. The in situ thermosensitive gel drug-loaded preparation according to claim 1, characterized in that: The mass percentage of polydopamine in the composition is 1%-5%.

6. The in situ thermosensitive gel drug-loaded preparation according to claim 1, characterized in that: The helical polyisocyanide polypeptide chain length in the composition is 0.5K-5K.

7. The in situ thermosensitive gel drug-loaded preparation according to claim 1, characterized in that: Dopamine hydrochloride is dissolved in 1x PBS at pH=8, and is self-polymerized into polydopamine under alkaline conditions, which is ready for use.

8. A method for preparing an in situ thermosensitive gel drug-loaded preparation, for preparing an in situ thermosensitive gel drug-loaded preparation as claimed in any one of claims 1 to 7, characterized in that: The method comprises the following steps: Dissolve Poloxamer 407 in ddH2O at low temperature to prepare a stock solution, and let it stand for a day and night to dissolve for later use; When the raw materials of the composition include poloxamer 188, the poloxamer 188 solution is dissolved in ddH2O at room temperature to prepare a stock solution for standby use; When the raw materials of the composition include hydroxypropyl methylcellulose, the hydroxypropyl methylcellulose is dissolved in ddH2O at room temperature to prepare a stock solution, and the solution is allowed to stand for one day and night to dissolve for later use; When the raw materials of the composition include polydopamine, dopamine hydrochloride is dissolved to self-polymerize into polydopamine under alkaline conditions, and the polydopamine is prepared and used immediately; When the raw materials of the composition include a helical polyisocyanate polypeptide, the helical polyisocyanate polypeptide is dissolved in 1x PBS to prepare a stock solution for later use; The raw materials required for the composition are uniformly mixed to obtain an in-situ thermosensitive gel drug-carrying preparation.

9. The method for preparing the in situ thermosensitive gel drug-loaded preparation according to claim 8, characterized in that: Hydroxypropyl methylcellulose was dissolved in ddH2O at 30-50°C.

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