A preparation method of an anti-epilepsy carbon dot suspension

Abstract

This invention, entitled "A Method for Preparing an Antiepileptic Carbon Dot Suspension," belongs to the field of pharmaceutical technology. It relates to a method for preparing a nanoparticle suspension and provides a carbon dot suspension for treating epilepsy. The raw materials and excipients of the suspension include: carbon dots from human hair, stabilizers, flavoring agents, and preservatives. The suspension of this invention exhibits stable properties, simple excipient composition and preparation process, and excellent taste. The most preferred formulation is: 5g of carbon dots from human hair, 0.125% CMC-Na, 0.05% ethylparaben, and 2.5% sucrose.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals and chemicals, specifically to an anti-epileptic carbon dot suspension and its preparation method. Background Technology

[0002] Epilepsy is a chronic neurological disorder characterized by recurrent seizures caused by abnormal synchronized discharges of neurons in the brain. It is the most common chronic disease of the central nervous system in children, who are more susceptible to epilepsy than adults. Currently, the main treatment for epilepsy is chemical drugs. While their effectiveness has been acknowledged, they suffer from serious side effects such as cognitive impairment, memory decline, and depression, as well as problems like rebound effects and drug tolerance upon discontinuation. Therefore, there is an urgent clinical need for new drugs with high efficacy and minimal side effects for epilepsy. Our team previously invented a traditional Chinese medicine for treating epilepsy (charred human hair). Given that the patient population is primarily children, a liquid formulation convenient for children to take is the most suitable choice.

[0003] Carbon dots from human hair (CC-CDs) have both good water solubility and good lipid solubility. With a particle size of <10nm, which is larger than that of general small molecule compounds, they are prone to sedimentation in solution, causing instability. Therefore, it is very important to select a suitable stabilizer.

[0004] The interaction between carbon dots and excipients is an important aspect that needs to be examined by those in the field. If the excipients are not selected appropriately, it will affect the efficacy of the carbon dots or even lead to their failure. Summary of the Invention

[0005] In view of the above-mentioned existing technical features, the purpose of this invention is to provide a carbon dot suspension for treating epilepsy with stable properties, simple excipient composition and preparation process, and excellent taste, as well as a method for preparing the same.

[0006] This invention provides a carbon dot suspension for treating epilepsy, comprising carbon dots and excipients, characterized in that the mass ratio of carbon dots to stabilizer is 1 to 10:1.

[0007] This invention provides a method for preparing a carbon dot suspension for treating epilepsy, characterized by comprising the following steps:

[0008] The preparation of anti-epileptic CC-CDs follows the method described in our previously filed patent (A Carbon Nanoparticle with Anti-epileptic Effect, Application No. 202211112401.7): Carbonized human hair is boiled in water to obtain an aqueous solution containing CC-CDs. The solution is further filtered, the filtrate is concentrated, and the concentrate is purified by dialysis or ultrafiltration to obtain a purified CC-CDs aqueous solution. After drying, CC-CDs powder is obtained. The CC-CDs have a particle size of 1-9 nm. The CC-CDs are composed of a carbon skeleton with a lattice spacing of approximately 0.2 nm, including but not limited to C, N, and O elements.

[0009] Weigh an appropriate amount of CC-CDs dry powder, add an appropriate amount of water and stir well to obtain a CC-CDs solution, or take an appropriate amount of freshly prepared CC-CDs solution, add a well-swollen stabilizer, preservative and flavoring agent and mix well to obtain the solution.

[0010] Preferably, in any of the above embodiments, the preservative is ethylparaben.

[0011] In any of the above embodiments, the flavoring agent is preferably sucrose.

[0012] The carbon dot suspension for treating epilepsy of the present invention has the following beneficial technical effects compared with the prior art:

[0013] This invention optimizes the drug-excipient formulation ratio of the suspension, selecting a ratio with good compatibility between the drug and excipients. It eliminates the need for special excipients, solving the problem of excipients easily interfering with the fluorescence intensity of carbon dots, thus causing unstable efficacy. It also addresses the issue of carbon dots easily settling and becoming unstable. The preparation method is simple and low-cost. The resulting product has stable properties, a simple excipient composition and preparation process, and excellent taste.

[0014] Experiment 1: Carbonized human hair + CMC-Na + Potassium sorbate + Sucrose

[0015] Preparation of CMC-Na solution: Measure 40 ml of deionized water into a beaker using a graduated cylinder, weigh 0.8 g of CMC-Na powder into the beaker, and stir slowly and continuously with a glass rod to allow it to fully swell, thus obtaining a 2% CMC-Na solution.

[0016] Take 5 ml of a 1 g / ml solution of carbonized human hair (based on the mass of carbonized human hair) into three 15 ml centrifuge tubes, and add 2.5 ml, 1.25 ml, and 0.625 ml of a 2% CMC-Na solution, respectively. Dilute to 10 ml with deionized water to obtain final mass fractions of 0.5%, 0.25%, and 0.125%. Weigh 0.01 g of potassium sorbate and 0.25 g of sucrose into the centrifuge tubes to obtain the final solution.

[0017] Experimental Example 2: Carbonized human hair + CMC-Na + Ethylparaben + Sucrose

[0018] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, and add 2.5 ml, 1.25 ml, and 0.625 ml of 2% CMC-Na solution respectively to make the final mass fractions 0.5%, 0.25%, and 0.125%. Make up to 10 ml with deionized water. Weigh 0.005 g of ethylparaben and 0.25 g of sucrose into the centrifuge tubes to obtain the final product.

[0019] Experimental Example 3: Carbonized human hair + CMC-Na + High-temperature sterilization + Sucrose

[0020] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, and add 2.5 ml, 1.25 ml, and 0.625 ml of 2% CMC-Na solution respectively to make the final mass fractions 0.5%, 0.25%, and 0.125%. Make up to 10 ml with deionized water, weigh 0.25 g of sucrose into the centrifuge tube, sonicate, and heat in a 100℃ water bath for 30 min to obtain the final product.

[0021] Experiment Example 4: Carbonized human hair + water-soluble starch + potassium sorbate + sucrose

[0022] Preparation of saturated starch solution: Measure 40 ml of deionized water into a beaker using a graduated cylinder, add sufficient starch solution into the beaker, stir thoroughly, centrifuge, and collect the supernatant to obtain the saturated starch solution.

[0023] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, and add 5 ml, 2.5 ml, and 1.25 ml of saturated starch solution respectively. Make up to 10 ml with deionized water to obtain three solutions of high, medium, and low concentrations. Weigh 0.01 g of potassium sorbate and 0.25 g of sucrose into centrifuge tubes to obtain the final solution.

[0024] Experimental Example 5: Carbonized human hair + water-soluble starch + ethylparaben + sucrose

[0025] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, and add 5 ml, 2.5 ml, and 1.25 ml of saturated starch solution respectively. Make up to 10 ml with deionized water to obtain three solutions of high, medium, and low concentrations. Weigh 0.005 g of ethylparaben and 0.25 g of sucrose into centrifuge tubes to obtain the final solution.

[0026] Experimental Example 6: Human hair charcoal + water-soluble starch + high-temperature sterilization + sucrose

[0027] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, add 5 ml, 2.5 ml, and 1.25 ml of saturated starch solution respectively, and make up to 10 ml with deionized water to obtain high, medium, and low concentration solutions. Weigh 0.25 g of sucrose into a centrifuge tube, sonicate, and heat in a 100°C water bath for 30 min to obtain the final solution.

[0028] Experiment 7: Carbonized human hair + Gelatin + Potassium sorbate + Sucrose

[0029] Preparation of gelatin solution: Measure 40 ml of deionized water into a beaker using a graduated cylinder, weigh 1 g of gelatin powder into the beaker, stir to dissolve, place in an 80℃ water bath to swell for 2 hours, and prepare a 2% gelatin solution for later use.

[0030] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, and add 1 ml, 0.5 ml, and 0.25 ml of 2% gelatin solution to each tube. Dilute to 10 ml with deionized water to obtain final mass fractions of 0.2%, 0.1%, and 0.05%, respectively. Weigh 0.01 g of potassium sorbate and 0.25 g of sucrose into the centrifuge tubes to obtain the final solution.

[0031] Experimental Example 8: Human hair charcoal + gelatin + ethylparaben + sucrose

[0032] Take 5 ml of 1 g / ml charred human hair solution into three 15 ml centrifuge tubes, and add 1 ml, 0.5 ml, and 0.25 ml of 2% gelatin solution respectively. Dilute to 10 ml with deionized water to make the final mass fractions 0.2%, 0.1%, and 0.05%. Weigh 0.005 g of ethylparaben and 0.25 g of sucrose into the centrifuge tubes to obtain the final solution.

[0033] Experimental Example 9: Human hair charcoal + gelatin + high-temperature sterilization + sucrose

[0034] Take 5 ml of 1 g / ml human hair charcoal solution into three 15 ml centrifuge tubes, add 1 ml, 0.5 ml, and 0.25 ml of 2% gelatin solution respectively, and make up to 10 ml with deionized water to make the final mass fractions 0.2%, 0.1%, and 0.05%. Weigh 0.25 g of sucrose into the centrifuge tubes, sonicate, and then incubate in a water bath at 100℃ for 30 min to obtain the final product.

[0035] Example: Stability Evaluation

[0036] The long-term stability of the nine samples prepared in the above experimental examples was investigated. The appearance was observed at month 0 and month 12, and the fluorescence intensity emitted by the samples was detected by a fluorescence spectrophotometer to investigate the stability.

[0037] (I) Appearance

[0038] As the storage time increased, the suspension with added potassium sorbate changed from a uniform and transparent state to show black mold spots and white precipitate; the suspension with added potassium sorbate changed from a uniform and transparent state to a pale yellow state; the suspension with added ethylparaben remained uniform and transparent. Therefore, ethylparaben was selected as the preservative.

[0039] (II) Fluorescence Measurement (Wavelength Selection: 355-460 nm)

[0040] 1. Determination of basic fluorescence intensity of excipients and CC-CDs

[0041] Take 200 μl of the excipient solution into each 96-well plate, turn on the fluorescence spectrophotometer, select the wavelength as 355-460 nm, set deionized water as the blank well to calibrate the instrument, and record the detection data, as shown in Table 1.

[0042] Table 1 Basic fluorescence values

[0043]

[0044]

[0045] As shown in Table 1, the fluorescence intensity of individual stabilizers (CMC-Na, starch, gelatin), individual preservatives (potassium sorbate, ethylparaben), individual flavoring agents (sucrose), and mixed excipients (stabilizer + preservative + flavoring agent) is consistent with that of the blank wells. Therefore, it is considered that the excipients have no fluorescent properties.

[0046] 2. Fluorescence intensity determination of CC-CDs suspension

[0047] Measurement results of fluorescence intensity in month 0

[0048] Take 200 μl of the suspension into each 96-well plate, set up 3 replicates for each well, turn on the fluorescence spectrophotometer, select the wavelength 355-460 nm, set deionized water as the blank well to calibrate the instrument, and record the detection data.

[0049] Table 2 Fluorescence values ​​in October

[0050]

[0051]

[0052] As shown in Table 2, the fluorescence intensity of carbonized human hair was 119.67. The fluorescence intensity of prescriptions numbered 003, 009, 017, and 027 was closest to that of carbonized human hair itself (±5).

[0053] The fluorescence intensity measurements at month 12 are shown in Table 3.

[0054] Table 3 Fluorescence values ​​in December

[0055]

[0056] Table 3 shows that the fluorescence intensity of carbonized human hair was 122.67, with prescriptions numbered 006, 009, 015, and 024 having fluorescence intensities closest to those of carbonized human hair (±5). Combining the results of the previous two measurements, prescription number 009 (0.125% CMC-Na + 0.05% ethylparaben + 0.25g sucrose + carbonized human hair) showed the best performance. The optimal prescription is: 5g carbonized human hair, 0.125% CMC-Na, 0.05% ethylparaben, and 2.5% sucrose.

Claims

1. A carbon dot suspension for treating epilepsy, comprising carbon dots and excipients, characterized in that, The mass ratio of carbon dots to stabilizers is 1 to 10:

1.

2. The method for preparing a carbon dot suspension for treating epilepsy as described in claim 1, characterized in that, Includes the following steps: Human hair charcoal is boiled in water to obtain an aqueous solution containing human hair charcoal carbon dots (CC-CDs). The solution is further filtered, and the filtrate is concentrated. The concentrate is purified by dialysis or ultrafiltration to obtain a purified CC-CDs aqueous solution, which is then dried to obtain CC-CDs powder. The CC-CDs have a particle size of 1-9 nm. The CC-CDs are composed of a carbon skeleton with a lattice spacing of about 0.2 nm, including but not limited to C, N, and O elements. Weigh an appropriate amount of CC-CDs dry powder, add an appropriate amount of water and stir well to obtain a CC-CDs solution, or take an appropriate amount of freshly prepared CC-CDs solution, add a well-swollen stabilizer, preservative and flavoring agent and mix well to obtain the solution. Preferably, in any of the above embodiments, the preservative is ethylparaben. In any of the above embodiments, the flavoring agent is sucrose.

3. The method for preparing a carbon dot suspension for treating epilepsy as described in claim 2, characterized in that: The flavoring agent mentioned in the steps is sucrose.

4. The method for preparing a carbon dot suspension for treating epilepsy as described in claim 2, characterized in that: The preservative is ethylparaben.

5. The method for preparing a carbon dot suspension for treating epilepsy as described in claim 2, characterized in that: The stabilizer is CMC-Na, water-soluble starch, or gelatin, with CMC-Na being the most preferred stabilizer.

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