A matrine sustained-release material and preparation method thereof

Abstract

The invention discloses a matrine controlled-release material and a preparation method thereof, wherein carbon nanotubes are used as carriers, matrine is used as a template molecule, methacrylic acid is used as a functional monomer, prepolymerization is first performed in an alkaline environment, and then a crosslinking agent and an initiator are added to react. After the reaction is completed, the template molecule matrine is washed away, and the reaction is dried to obtain a surface molecular imprinting polymer of matrine. After the surface molecular imprinting polymer saturates with matrine solution, the matrine controlled-release material is obtained. The invention can simply prepare a controlled-release material with a "memory" function, which can stably and permanently release matrine and inhibit the vitality of liver cancer cells.

Description

Technical Field

[0001] The invention belongs to the field of controlled-release materials, and in particular relates to a matrine controlled-release material and a preparation method thereof. Background Art

[0002] Matrine (MAT) belongs to the tetracyclic quinolizine class of compounds and can be regarded as a fusion of two quinolizine rings. It has the effects of anti-liver damage, anti-liver fibrosis, anti-tumor, anti-inflammatory and anti-allergic, and protective effects on the cardiovascular system. It is now widely used as an injection for the treatment of hepatitis and anti-tumor. At present, matrine injection is mainly used for intravenous injection and intravenous drip in clinical application. Although injection has the advantages of rapid efficacy, reliable effect and long-term effect, its defects cannot be avoided: (1) High quality requirements. Usually, the injection solution needs to be tested for pyrogens in accordance with the provisions of SFDA standard YBH06592006. The rabbit method and the horseshoe crab reagent method are commonly used to test bacterial endotoxins. These two detection methods have high accuracy but are inconvenient to operate. It is impossible to quickly obtain test results and control the quality of intermediates; (2) The preparation technology and production process are complex and the production cost is high; (3) It is inconvenient to use and painful to inject; (4) The physiological effects are difficult to reverse and the risk is relatively high. Clinical accidents caused by injections should not be underestimated. Therefore, researchers hope to find a new way of administering the drug to avoid these problems.

[0003] In recent years, the nano drug delivery system made of drugs and carrier materials using nanotechnology is quite active. The particle size is mostly between 1nm-1000nm. Through a series of chemical modifications or loading, the drug can be encapsulated in the nano material or linked to the surface or inside of the nano material, which can achieve the purpose of specific delivery of the drug to the target area, and can be triggered to release the drug under certain conditions. Molecular Imprinting Technology (MIT) is a marginal discipline branch developed in recent years that integrates the advantages of many disciplines such as polymer synthesis, molecular design, molecular recognition, and biomimetic bioengineering. Molecular Imprinted Polymer (MIP) prepared based on this technology has the characteristics of high affinity and selectivity with template molecules, strong resistance to harsh environments, good stability, long service life, and a wide range of applications. Because MIP has three-dimensional holes for template drugs, the sites on its surface can achieve reversible binding with template molecules through different experimental conditions, thereby improving the therapeutic effect of drugs while reducing their toxic side effects. MIT will have broad application prospects in the targeted delivery of target ingredients to drugs. Among them, the emulsion polymerization method can design and prepare polymer microspheres with suitable morphology and size, which have great potential as nano drug delivery systems. Summary of the invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a matrine sustained-release material and a preparation method thereof, which can effectively control the release rate of matrine, has a simple preparation process, is easy to use and has low cost.

[0005] The purpose of the present invention is achieved through the following technical solutions:

[0006] A method for preparing a matrine controlled-release material comprises the following steps: using carbon nanotubes as carriers, matrine as a template molecule, and methacrylic acid as a functional monomer, prepolymerizing the materials in an alkaline environment, and then adding a crosslinking agent and an initiator to react. After the reaction is completed, the template molecule matrine is washed away, and the materials are dried to obtain a surface molecular imprinted polymer of matrine. After the surface molecular imprinted polymer is saturatedly adsorbed with a matrine solution, the matrine controlled-release material is obtained.

[0007] The carbon nanotubes are multi-walled carbon nanotubes.

[0008] The molar ratio of the functional monomer to the template molecule is 1:3 to 1:7.

[0009] The alkaline environment is a Tris-HCl buffer solution, and the pH is adjusted to pH 8-10.

[0010] The prepolymerization is to dissolve the carbon nanotubes, matrine and methacrylic acid in Tris-HCl buffer by ultrasonication for 10 to 30 minutes, and then stir at a constant temperature of 20 to 45 degrees Celsius for 2 to 6 hours.

[0011] The cross-linking agent and initiator are dopamine (DA); the molar ratio of dopamine to the functional monomer is 3:1 to 1:3.

[0012] The conditions for adding the crosslinking agent and the initiator to react are: at room temperature, magnetic stirring polymerization reaction for 6h to 24h.

[0013] The template molecule matrine is washed away by taking out the reactant, washing it with methanol, then eluting it with a mixed solution of methanol and glacial acetic acid (volume ratio of 4:1 to 9:1) in an oscillator for 36 hours to 48 hours, and then washing it with methanol and water respectively until it is neutral.

[0014] The concentration of the matrine solution is 80-160 μ6 / mL.

[0015] The adsorption time is 1 h to 3 h.

[0016] A matrine sustained-release material is prepared by adopting the above steps.

[0017] Compared with the prior art, the present invention has the following advantages and effects:

[0018] (1) The present invention uses matrine as a template to synthesize the molecular imprinting polymer on the surface of multi-walled carbon nanotubes. The molecular imprinting polymer has a certain recognition of matrine and can adsorb matrine. Moreover, the adsorption is reversible. After saturated adsorption of the obtained molecular imprinting polymer, it is used to inhibit the activity of liver cancer cells.

[0019] (2) The preparation process of the present invention is simple, the Tris-HCl buffer solvent is non-toxic and harmless, and the multi-walled carbon nanotube carrier and dopamine used as a cross-linking agent and initiator have good biocompatibility, so the sustained-release process is safer and more environmentally friendly, and can stably and long-term release matrine.

[0020] (3) The present invention can simply prepare a sustained-release material with a "memory" function, which can stably and permanently release matrine and inhibit the activity of liver cancer cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The figure is a schematic diagram of the process of synthesizing surface molecular imprinted polymers according to the present invention.

[0022] Figure 2 This is a scanning electron microscope image of the surface molecular imprinted polymer after drug loading of the present invention.

[0023] Figure 3 This is a graph of the in vitro release rate of the drug-loaded material of the present invention.

[0024] Figure 4 This is a graph showing the drug-loaded material of the present invention inhibiting the activity of liver cancer cells. DETAILED DESCRIPTION

[0025] In order to facilitate the understanding of the present invention, the present invention will be described in detail below in conjunction with specific embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be pointed out that, for those skilled in the art, without departing from the inventive concept, the present invention can also make some variations and improvements, which all belong to the protection scope of the present invention.

[0026] Example 1

[0027] Weigh the template molecule and the functional monomer MAA in a molar ratio of 1:3 and mix them in a conical flask, add multi-walled carbon nanotubes, and then add solvent, with 25 mL of solvent corresponding to each gram of multi-walled carbon nanotubes; ultrasonically dissolve for 30 minutes, add a magnetic stirring rotor, seal the bottle mouth, stir at a constant temperature of 25°C for about 2 hours, then open the bottle stopper, add dopamine as a cross-linking agent and initiator, with 0.6 mmol of dopamine corresponding to each millimole of functional monomer MAA; ultrasonically stir, seal the bottle mouth, and stir the polymerization reaction at a constant temperature for 6 hours, take out the polymer, wash it with methanol, elute it with a mixed solution of methanol and glacial acetic acid (volume ratio of 4:1) in a constant temperature oscillator for 48 hours, wash it with methanol and water until neutral, respectively, to obtain a surface molecular imprinting polymer of matrine; then use 100 mg of the surface molecular imprinting polymer to adsorb 100 μg / mL of matrine solution for 1 hour, wash and dry, and obtain a matrine sustained-release material. Figure 1 As shown in FIG. 1 , it is a schematic diagram of the process of synthesizing matrine molecular imprinting polymer using matrine as a template, including the synthesis, desorption and adsorption stages of the polymer. Figure 2 The figure shows an electron microscope scanning image (x10000) of the matrine molecular imprinted polymer prepared by the present invention. It can be seen from the figure that the surface of the multi-walled carbon nanotube is rough and has obvious attachments, indicating that the molecular imprinted polymer has been connected.

[0028] Example 2

[0029] The template molecule and the functional monomer MAA in a molar ratio of 1:4 were weighed and mixed in a conical flask, multi-walled carbon nanotubes were added, and then a solvent was added, with each gram of multi-walled carbon nanotubes corresponding to 20 mL of solvent; ultrasonic dissolution was performed for 30 minutes, a magnetic stirring rotor was added, the bottle mouth was sealed, and the bottle stopper was opened after constant temperature stirring at 35°C for about 2 hours, and dopamine was added as a cross-linking agent and initiator, wherein each millimole of the functional monomer MAA corresponds to 0.4 mmol of dopamine; ultrasonic stirring was performed, the bottle mouth was sealed, and the polymerization reaction was stirred at a constant temperature for 12 hours, the polymer was taken out, washed with methanol, and eluted with a mixed solution of methanol and glacial acetic acid (volume ratio of 4:1) in a constant temperature oscillator for 72 hours, and washed with methanol and water to neutrality, respectively, to obtain a surface molecular imprinting polymer of matrine; then 100 mg of the surface molecular imprinting polymer was used to adsorb 100 μg / mL of matrine solution for 1 hour, and after washing and drying, a matrine sustained-release material was obtained.

[0030] Example 3

[0031] The template molecule and the functional monomer MAA in a molar ratio of 1:5 are weighed and mixed in a conical flask, multi-walled carbon nanotubes are added, and then a solvent is added, with each gram of multi-walled carbon nanotubes corresponding to 20 mL of solvent; ultrasonic dissolution is performed for 30 minutes, a magnetic stirring rotor is added, the bottle mouth is sealed, and the bottle stopper is opened after constant temperature stirring at 25°C for about 2 hours, and dopamine is added as a cross-linking agent and initiator, wherein each millimole of the functional monomer MAA corresponds to 0.8 mmol of dopamine; ultrasonic stirring is performed, the bottle mouth is sealed, and the polymerization reaction is performed at a constant temperature for 6 hours, the polymer is taken out, washed with methanol, and eluted with a mixed solution of methanol and glacial acetic acid (volume ratio of 4:1) in a constant temperature oscillator for 96 hours, and washed with methanol and water to neutrality, respectively, to obtain a surface molecular imprinting polymer of matrine; then 100 mg of the surface molecular imprinting polymer is used to adsorb 100 μg / mL of matrine solution for 1 hour, and after washing and drying, a matrine sustained-release material is obtained.

[0032] Test Example 1: In vitro release test

[0033] 30 mg of the matrine sustained-release material prepared in the present invention was dissolved in 50 mL of PBS (pH 4-7), the matrine content was measured at regular intervals, and the in vitro release rate was calculated. Figure 3 This is a graph of the release rate of matrine sustained-release material at different pH values ​​in vitro. It can be seen from the figure that at pH 4, the release rate reaches 83.72%, and the release rate decreases as the pH increases. This provides a theoretical basis for achieving sustained-release effects due to different release rates in different organs and parts of the body due to different environments.

[0034] Test Example 2: Effect of matrine sustained-release material on the viability of liver cancer cells

[0035] The matrine sustained-release material (MAT-loaded MTPs) prepared by the present invention was co-incubated with liver cancer cells HepG2 at the same concentration, with matrine MAT (concentration of 160 μg / mL) as a control group, for 96 h. The cell morphology and number were observed under an inverted microscope, and the matrine content in the culture medium was measured by high performance liquid chromatography, and the inhibitory abilities of the matrine sustained-release material and the matrine group on liver cancer cells were compared. Figure 4 This is a graph showing that matrine sustained-release material inhibits the activity of liver cancer cells (PBS is a phosphate buffer control group). It can be seen from the graph that compared with the control matrine solution group, the number of cells in the matrine sustained-release material group decreased more with the extension of time, that is, the matrine sustained-release material of the present invention has a stronger inhibitory ability on liver cancer cells than the matrine solution.

[0036] The above descriptions are merely embodiments of the present invention, but the implementation methods of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention are equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for preparing a matrine sustained-release material, characterized in that: The method uses carbon nanotubes as carriers, matrine as template molecules, and methacrylic acid as functional monomers, and prepolymerizes in an alkaline environment, then adds a crosslinking agent and an initiator to react. After the reaction is completed, the template molecule matrine is washed away, and the reaction is dried to obtain a surface molecular imprinted polymer of matrine. After the surface molecular imprinted polymer is saturated with the matrine solution, the matrine sustained-release material is obtained. The molar ratio of the functional monomer to the template molecule is 1:3~1:7; the alkaline environment is a Tris-HCl buffer, and the pH is adjusted to pH8~10; the prepolymerization is to ultrasonically dissolve the carbon nanotubes, matrine, and methacrylic acid in a Tris-HCl buffer for 10 min~30 min, and then stir at a constant temperature of 20°C~45°C for 2~6 h; the cross-linking agent and initiator are dopamine; the molar ratio of dopamine to the functional monomer is 3:1~1:3; the conditions for adding the cross-linking agent and initiator to react are: magnetic stirring polymerization reaction at room temperature for 6 h~24 h; the template molecule matrine is washed away, and the reactant is taken out, washed with methanol, and then eluted with a mixed solution of methanol and glacial acetic acid in an oscillator for 36 h~48 h, and then washed with methanol and water respectively until neutral.

2. The method for preparing the matrine sustained-release material according to claim 1, characterized in that: The carbon nanotubes are multi-walled carbon nanotubes.

3. The method for preparing the matrine sustained-release material according to claim 1, characterized in that: The concentration of the matrine solution is 80-160 μg / mL, and the adsorption time is 1-3 hours.

4. A matrine sustained-release material, characterized in that: The method is prepared by any one of claims 1 to 3.

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