Preparation method of porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride
The preparation of porous carbon drug carriers by high-temperature carbonization treatment of porous polymers has been solved, and the biocompatibility and solubility problems have been achieved, efficient adsorption and release of drugs have been achieved, and the biosafety and solubility of drugs have been improved.
Patent Information
- Application Number
- CN202211029107.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-08-26
- Publication Date
- 2025-08-12
- Estimated Expiration
- 2042-08-26
AI Technical Summary
The existing porous organic polymer drug carriers have defects in biocompatibility, limiting their application potential in organisms, and the low solubility of insoluble drugs in vivo limits their clinical use.
Porous carbon materials are prepared by high-temperature carbonization of the precursor porous polymer, and berberine hydrochloride is loaded thereon to form a porous carbon drug carrier to achieve efficient adsorption and release of the drug.
It improves the biosafety and solubility of the drug, and after drug loading, it changes from crystalline state to amorphous morphology, enhancing the stability and release speed of the drug.
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Figure CN115554407B_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of pharmaceutical carriers, and in particular relates to a method for preparing a porous carbon drug carrier capable of efficiently adsorbing and releasing berberine hydrochloride. Background Art
[0002] In clinical applications, the low solubility of some poorly soluble drugs in the body greatly limits their application. Loading poorly soluble drugs on some carriers can increase drug solubility, thereby improving the bioavailability of poorly soluble drugs, which is an effective measure to solve the limited clinical use of poorly soluble drugs. With the rapid development of drug carrier systems, it has been found that loading poorly soluble drugs into porous organic polymers (POPs) can effectively improve the solubility of drugs. This is because POPs can disperse poorly soluble drugs well due to their high specific surface area, rich pores and pore confinement, effectively reducing the crystallinity of poorly soluble drugs and thus improving their solubility.
[0003] However, many synthetic POP materials suffer from biocompatibility deficiencies and are not biocompatible, severely limiting their potential for practical application. High-temperature carbonization is an effective method for removing volatile substances from materials and also plays a role in regulating the pore structure of materials, particularly improving the biosafety of drug carriers. Therefore, it is often used to prepare unique porous carbon drug carriers. Summary of the Invention
[0004] The present invention provides a method for preparing a porous carbon drug carrier that efficiently adsorbs and releases berberine hydrochloride. By subjecting a precursor porous polymer to high-temperature carbonization treatment, a porous carbon material is successfully prepared and used for the loading and release of the poorly soluble drug berberine hydrochloride.
[0005] The purpose of the present invention is achieved through the following technical solutions:
[0006] A method for preparing a porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride comprises the following steps: dissolving 4,4′-(bromomethyl)biphenyl and 1,3,5-tris(bromomethyl)benzene in 1,2-dichloroethane under a nitrogen atmosphere, rapidly adding anhydrous ferric chloride, stirring at 45°C for 1 hour, heating to 80°C and continuing stirring for 1 hour, cooling to room temperature, filtering, washing with methanol, and vacuum drying at 70°C for 24 hours to obtain a precursor polymer PBTP; placing the precursor polymer PBTP in a porcelain boat, placing it in a tube furnace, and calcining it under a nitrogen atmosphere to obtain a porous carbon material PBTC; ultrasonically dissolving berberine hydrochloride in an appropriate amount of methanol, adding the porous carbon material PBTC, sealing the reaction vessel, magnetically stirring for 24 hours, and then drying. After the methanol evaporates, a BBH porous carbon drug carrier is obtained.
[0007] Furthermore, the molar ratio of the 4,4′-(bromomethyl)biphenyl: the 1,3,5-tris(bromomethyl)benzene is 1:4 to 4:1, and the molar ratio of the 1,3,5-tris(bromomethyl)benzene: the 1,2-dichloroethane: the anhydrous ferric chloride is 4:40:5.
[0008] Furthermore, the mass ratio of the berberine hydrochloride to the porous carbon material PBTC is 1:1.
[0009] Furthermore, the specific process of calcining under N2 atmosphere is as follows: in N2 atmosphere, heating the tube furnace to 500-1200°C at a heating rate of 5-20°C / min, keeping the temperature for 2-5 hours and then cooling to room temperature to obtain the porous carbon material PBTC.
[0010] Furthermore, the specific process of calcining under N2 atmosphere is as follows: in N2 atmosphere, the temperature of the tube furnace is raised to 1000°C at a heating rate of 10°C / min, kept at this temperature for 2 hours, and then cooled to room temperature to obtain the porous carbon material PBTC.
[0011] Beneficial effects of the present invention:
[0012] 1. A simple high-temperature carbonization treatment of porous organic polymers was used to successfully remove the biotoxic substance Br, significantly improving the biosafety of the carrier material.
[0013] 2. The specific surface area and pore size distribution of porous carbon materials are regulated by changing the carbonization temperature.
[0014] 3. During the drug loading process, the crystalline state is transformed into the amorphous state, which greatly improves the solubility of the drug. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] Figure 1 : Infrared spectra of the berberine hydrochloride raw material BBH, the porous carbon material PBTC-1000, and the berberine hydrochloride porous carbon drug carrier PBTC-1000 / BBH in this Example 1.
[0016] Figure 2 : SEM images of berberine hydrochloride bulk drug BBH (a) and berberine hydrochloride porous carbon drug carrier PBTC-1000 / BBH (b) in Example 1.
[0017] Figure 3 : In vitro dissolution curves of the berberine hydrochloride porous carbon drug carrier PBTC-1000 / BBH and BBH in Example 1 in (a) pH 7.4 phosphate buffer solution, (b) pH 6.8 phosphate buffer solution, and (c) water; (d) in vitro dissolution curves of PBTC-1000 / BBH under different solvent conditions.
[0018] Figure 4 : UV full wavelength scanning spectra of BBH under different influencing factors in Example 1: (a) temperature, (b) light and (c) humidity.
[0019] Figure 5 : Effects of different concentrations of berberine hydrochloride raw material BBH (a), porous carbon material PBTC-1000 (b), and berberine hydrochloride porous carbon drug carrier PBTC-1000 / BBH (c) on cell viability in this Example 1. DETAILED DESCRIPTION
[0020] In order to further understand the present invention, preferred embodiments of the present invention are described below in conjunction with examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, rather than limiting the claims of the present invention.
[0021] The reagents and instruments used in the present invention without manufacturer indication are all conventional products that can be purchased from the market.
[0022] Example 1
[0023] The precursor polymer PBTP was synthesized by dissolving 0.01 mol of 4,4′-(bromomethyl)biphenyl (BBMBP) and 0.04 mol of 1,3,5-tris(bromomethyl)benzene (TBB) in 4 mol of 1,2-dichloroethane under a nitrogen atmosphere. 0.05 mol of anhydrous ferric chloride was quickly added to the solution, stirred at 45°C for 1 hour, then heated to 80°C and stirred rapidly for 1 hour. After the reaction, the mixture was cooled to room temperature, filtered, washed with methanol, and dried under vacuum at 70°C for 24 hours to obtain a dark brown solid powder, the precursor polymer PBTP.
[0024] High-temperature carbonization of the precursor polymer PBTP to prepare the porous carbon material PBTC: An appropriate amount of the precursor material PBTP was weighed into a porcelain boat and placed in a tube furnace. Under a nitrogen atmosphere, the sample was heated to 1000°C at a rate of 10°C / min. The temperature was maintained for 2 hours before being cooled to room temperature to obtain the porous carbon material PBTC, designated PBTC-1000.
[0025] Loading of berberine hydrochloride (BBH): 10 mg of berberine hydrochloride was dissolved in 5 mL of methanol and completely dissolved by ultrasound. Then, 10 mg of porous carbon material PBTC was dispersed in the above solution. The reaction vessel was sealed and the drug was evenly dispersed in the porous carbon material PBTC under the action of a magnetic stirrer. After stirring for 24 hours, the loaded solution was placed in a 35°C oven for drying. After the methanol evaporated, the BBH porous carbon drug carrier was obtained, which was recorded as PBTC-1000 / BBH.
[0026] Example 2
[0027] The precursor polymer PBTP was synthesized by dissolving 0.16 mol of 4,4′-(bromomethyl)biphenyl (BBMBP) and 0.04 mol of 1,3,5-tris(bromomethyl)benzene (TBB) in 4 mol of 1,2-dichloroethane under a nitrogen atmosphere. 0.05 mol of anhydrous ferric chloride was quickly added to the solution, stirred at 45°C for 1 hour, then heated to 80°C and stirred rapidly for 1 hour. After the reaction, the mixture was cooled to room temperature, filtered, washed with methanol, and dried under vacuum at 70°C for 24 hours to obtain a dark brown solid powder, the precursor polymer PBTP.
[0028] High-temperature carbonization of the precursor polymer PBTP to prepare the porous carbon material PBTC: An appropriate amount of the precursor material PBTP was weighed into a porcelain boat and placed in a tube furnace. Under a nitrogen atmosphere, the sample was heated to 1200°C at a rate of 20°C / min. The temperature was maintained for 2 hours before being cooled to room temperature to obtain the porous carbon material PBTC, designated PBTC-1200.
[0029] Loading of berberine hydrochloride (BBH): 10 mg of berberine hydrochloride was dissolved in 5 mL of methanol and completely dissolved by ultrasound. Then, 10 mg of porous carbon material PBTC was dispersed in the above solution. The reaction vessel was sealed and the drug was evenly dispersed in the porous carbon material PBTC under the action of a magnetic stirrer. After stirring for 24 hours, the loaded solution was placed in a 35°C oven for drying. After the methanol evaporated, the BBH porous carbon drug carrier was obtained, which was recorded as PBTC-1200 / BBH.
[0030] Example 3
[0031] The precursor polymer PBTP was synthesized by dissolving 0.04 mol of 4,4′-(bromomethyl)biphenyl (BBMBP) and 0.04 mol of 1,3,5-tris(bromomethyl)benzene (TBB) in 4 mol of 1,2-dichloroethane under a nitrogen atmosphere. 0.05 mol of anhydrous ferric chloride was quickly added to the solution, stirred at 45°C for 1 hour, then heated to 80°C and stirred rapidly for 1 hour. After the reaction, the mixture was cooled to room temperature, filtered, washed with methanol, and dried under vacuum at 70°C for 24 hours to obtain a dark brown solid powder, the precursor polymer PBTP.
[0032] High-temperature carbonization of the precursor polymer PBTP to prepare the porous carbon material PBTC: An appropriate amount of the precursor material PBTP was weighed into a porcelain boat and placed in a tube furnace. Under a nitrogen atmosphere, the sample was heated to 500°C at a rate of 5°C / min, held for 5 hours, and then cooled to room temperature to obtain the porous carbon material PBTC, designated PBTC-500.
[0033] Loading of berberine hydrochloride (BBH): 10 mg of berberine hydrochloride was dissolved in 5 mL of methanol and completely dissolved by ultrasound. Then, 10 mg of porous carbon material PBTC was dispersed in the above solution. The reaction vessel was sealed and the drug was evenly dispersed in the porous carbon material PBTC under the action of a magnetic stirrer. After stirring for 24 hours, the loaded solution was placed in a 35°C oven for drying. After the methanol evaporated, the BBH porous carbon drug carrier was obtained, which was recorded as PBTC-500 / BBH.
[0034] The BBH porous carbon drug carriers prepared in Examples 1-3 were subjected to an in vitro drug dissolution test:
[0035] Accurately weigh 5 mg of drug or a drug-carrier complex containing the same amount of drug into a dialysis bag (MD44-3500) sealed at one end. Add 5 mL of the corresponding medium and seal the other end. Place the dialysis bag in 200 mL of water, pH 6.8, or pH 7.4, respectively, and use magnetic stirring to ensure uniform drug diffusion. At 5, 10, 20, 30, 50, 70, 100, 120, 150, 180, 210, 240, 280, 320, 360, 400, and 450 min, 5 mL of medium was sampled from each system and the same amount of medium was added to the beaker simultaneously. The absorbance was measured using UV-visible spectrophotometry at a wavelength of 345 nm. The in vitro drug release at different sampling times was calculated according to formula (1).
[0036] Formula (1):
[0037] Q i : Drug dissolution at the time of sampling for the i-th time
[0038] C i : Drug concentration at the i-th sampling point
[0039] V1: volume of dissolution medium
[0040] V2: Sampling volume
[0041] n: number of sampling time points
[0042] M: total amount of drug
[0043] The BBH porous carbon drug carriers prepared in Examples 1-3 were subjected to drug stability experiments:
[0044] (1) Temperature effect experiment: 5 mg of BBH porous carbon drug carrier was accurately weighed and placed in two equal parts in an open watch glass, and then placed in an oven at 60°C for 10 days. Samples were taken on the 5th and 10th days, respectively. The samples were dissolved in methanol and diluted to a 100 mL volumetric flask. After ultrasonic treatment for 30 min, the drug loaded on the carrier was completely released. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and scanned at a wavelength of 345 nm using UV-visible spectrophotometry. The drug stability was determined by comparing the UV-visible full-wavelength scanning curve with that of BBH.
[0045] (2) Light effect experiment: 5 mg of BBH porous carbon drug carrier was accurately weighed and placed in two equal parts in an open watch glass. The drug carrier was placed under a light intensity of 4500 lx ± 500 lx for 10 days. Samples were taken on the 5th and 10th days respectively. The samples were dissolved in methanol and diluted to a volume of 100 mL in a volumetric flask. After ultrasonic treatment for 30 minutes, the drug loaded in the carrier was completely released. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and scanned by UV-visible spectrophotometry at a wavelength of 345 nm. The drug stability was determined by comparing the UV-visible full-wavelength scanning curve with that of BBH.
[0046] (3) Humidity effect experiment: 5 mg of BBH porous carbon drug carrier was accurately weighed and placed in two equal portions in an open watch glass. The mixture was placed at room temperature under a relative humidity of 75 ± 5% (saturated NaCl) for 10 days. Samples were taken on the 5th and 10th days, respectively. The samples were dissolved in methanol and diluted to a volume of 100 mL in a volumetric flask. After ultrasonic treatment for 30 min, the drug loaded on the carrier was completely released. The supernatant was filtered through a 0.22 μm ultrafiltration membrane and scanned at a wavelength of 345 nm using UV-visible spectrophotometry. The drug stability was determined by comparing the UV-visible full-wavelength scan curve with that of BBH.
[0047] The BBH porous carbon drug carriers prepared in Examples 1-3 were subjected to biocompatibility experiments:
[0048] To investigate the biocompatibility of the carrier materials, cytotoxicity tests were conducted using HepG2 cells on BBH, the porous carbon material PBTC, and the BBH porous carbon drug carrier. The results showed that the porous carbon material PBTC exhibited minimal cytotoxicity, maintaining high cell viability even at a high concentration of 1000 μg / mL. After drug loading, the BBH porous carbon drug carrier was used at concentrations of 0-100 μg / mL. The cytotoxicity at high concentrations was primarily due to the berberine hydrochloride itself. Therefore, the porous carbon material PBTC is a drug carrier with a stable structure and excellent biocompatibility.
[0049] from Figures 1 to 5It can be seen that in the present invention, the porous carbon material PBTC was successfully prepared by high-temperature carbonization treatment of the precursor polymer PBTP and was used for the loading and release of the poorly soluble drug berberine hydrochloride.
[0050] It was characterized by FTIR and SEM, and the results showed that berberine hydrochloride could be successfully loaded on the porous carbon material PBTC. Further characterization showed that the loaded berberine hydrochloride changed from a crystalline state to an amorphous state, which would be beneficial to the improvement of the solubility of berberine hydrochloride.
[0051] In vitro drug release experiments showed that compared to the API berberine hydrochloride, the loaded BBH porous carbon drug carrier exhibited significantly improved drug release rate and extent. This was primarily attributed to the drug's crystal transformation after loading. The amorphous form of berberine hydrochloride, free from lattice constraints, has a high free energy, significantly enhancing drug solubility. In experiments examining factors affecting drug stability, the full-wavelength UV spectrum of berberine hydrochloride from the BBH porous carbon drug carrier after exposure to high temperature, high humidity, and strong light was consistent with that of the API, demonstrating the drug's excellent stability under these conditions.
[0052] In examining the biocompatibility of carrier materials, HepG2 cells were used to conduct cytotoxicity tests on BBH, porous carbon material PBTC, and BBH porous carbon drug carriers. The results showed that the cytotoxicity of the carrier material PBTC-1000 was very weak, and the cells had high activity even at a high concentration of 1000 μg / mL. After drug loading, the drug-carrier complex PBTC-1000 / BBH was used at a concentration of 0-100 μg / mL. The cytotoxicity at high concentrations mainly came from berberine hydrochloride itself. Therefore, the porous carbon material PBTC-1000 is a type of drug-carrying material with stable structure and good biocompatibility.
[0053] Based on the disclosure of the above description, those skilled in the art may also make appropriate changes and modifications to the above embodiments. Therefore, the present invention is not limited to the specific embodiments disclosed and described above, and any modifications and variations of the present invention should also fall within the scope of protection of the claims of the present invention. In addition, although certain specific terms are used in this description, these terms are merely for convenience of description and do not constitute any limitation to the present invention.
Claims
1. A method for preparing a porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride, characterized in that: Under N2 atmosphere, 4,4′-(bromomethyl)biphenyl and 1,3,5-tri(bromomethyl)benzene were dissolved in 1,2-dichloroethane, and anhydrous ferric chloride was quickly added. After stirring at 45°C for 1 hour, the temperature was raised to 80°C and stirring was continued for 1 hour. The temperature was lowered to room temperature, filtered, washed with methanol, and dried in vacuum at 70°C for 24 hours to obtain the precursor polymer PBTP. The precursor polymer PBTP was placed in a porcelain boat, placed in a tube furnace, and calcined under a N2 atmosphere to obtain the porous carbon material PBTC; Berberine hydrochloride was ultrasonically dissolved in an appropriate amount of methanol, and the porous carbon material PBTC was added. The reaction vessel was sealed, and magnetic stirring was performed for 24 hours before drying. After the methanol evaporated, the BBH porous carbon drug carrier was obtained.
2. The method for preparing a porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride according to claim 1, characterized in that: The molar ratio of the 4,4′-(bromomethyl)biphenyl to the 1,3,5-tris(bromomethyl)benzene is 1:4 to 4:1, and the molar ratio of the 1,3,5-tris(bromomethyl)benzene to the 1,2-dichloroethane to the anhydrous ferric chloride is 4:40:
5.
3. The method for preparing a porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride according to claim 1, characterized in that: The mass ratio of the berberine hydrochloride to the porous carbon material PBTC is 1:
1.
4. The method for preparing a porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride according to claim 1, characterized in that: The specific process of calcining under N2 atmosphere is as follows: in N2 atmosphere, heating the tube furnace to 500-1200°C at a heating rate of 5-20°C / min, keeping the temperature for 2-5 hours and then cooling to room temperature to obtain the porous carbon material PBTC.
5. The method for preparing a porous carbon drug carrier for efficient adsorption and release of berberine hydrochloride according to claim 4, characterized in that: The specific process of calcining under N2 atmosphere is as follows: in N2 atmosphere, heating the tube furnace to 1000°C at a heating rate of 10°C / min, keeping the temperature for 2 hours and then cooling to room temperature to obtain the porous carbon material PBTC.