CN111773209A - Application of psoralen in preparation of medicine for preventing or treating cochlear hair cell injury - Google Patents
Application of psoralen in preparation of medicine for preventing or treating cochlear hair cell injury Download PDFInfo
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
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- A61K9/5042—Cellulose; Cellulose derivatives, e.g. phthalate or acetate succinate esters of hydroxypropyl methylcellulose
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- A61P27/16—Otologicals
Abstract
The invention discloses an application of psoralen in preparing a medicament for preventing or treating cochlear hair cell injury, wherein the medicament is an auricular point patch for preventing noise hearing injury; the auricular point patch bean is a psoralen hydrogel auricular point patch bean, wherein the psoralen hydrogel auricular point patch bean is obtained by sticking psoralen drug-loaded pellets on psoralen hydrogel and drying in the shade at room temperature for 24 hours; the effective components of the psoralen hydrogel auricular point patch bean are derived from kidney-tonifying traditional Chinese medicine bone fat supplement, the psoralen hydrogel auricular point patch bean is prepared by combining with the traditional auricular point treatment means of the traditional Chinese medicine, and pharmacodynamics verifies that the preparation has a targeted protection effect on cochlear cell injury caused by various reasons, can press and stimulate auricular points, can stably and effectively absorb the medicine through skin, and jointly protects hearing.
Description
(I) technical field
The invention relates to an auricular point sticking bean which can prevent auricular injury caused by recreational noise.
(II) background of the invention
Noise-induced hearing impairment is also known as noise-induced deafness, which is a slow, progressive sensorineural deafness caused by prolonged exposure to noise stimuli. The entertainment noise damage refers to ear damage caused by unreasonable use of audio equipment with earphones such as mobile phones and MP3, and entertainment places with high noise such as KTV and bars. Domestic and foreign research shows that serious damage to cochlear hair cells can be caused by long-time exposure to recreational noise with large volume. While each individual has a fixed number of cochlear hair cells, which have no regenerative capacity and, once lost, can cause irreversible damage to the user's hearing. Excessive hearing loss will have great impact on our individual and society, including physical and functional, social and emotional, and economic aspects.
Many acupoints, called ear acupoints for short, are densely distributed on the auricle. The acupoint is stimulated by pressing the ear acupoint at a fixed point, the physical information is transmitted to the corresponding viscera by the 'channels and collaterals', the operation function of the viscera is adjusted, and the functional adjustment is performed to defend the human body, so that the effects of preventing and treating diseases are achieved. The seed sticking and the magnetic bead sticking of the cowherb seeds are the prototype of the invention, the seeds are mainly stuck on the ear epidermis, the invention is relatively safe, and the invention has the health care and auxiliary treatment effects on various diseases such as insomnia, hypertension, angina after coronary heart disease operation, tinnitus and the like through the clinical practice of Chinese medicine for hundreds of years. However, the dosage form improvement and deep health care mechanism of the auricular point patch are not well researched and developed. The existing ear acupuncture point therapy only stimulates acupuncture points by physical means such as 'pressing' and 'magnetism', and does not combine effective medicine percutaneous absorption for treatment.
Fructus Psoraleae (Psoralea corilifolia Linn) is dry mature fruit of Psoralea corylifolia of Leguminosae, is a common clinical Chinese medicine, has warm nature and bitter taste, enters kidney and spleen channels, and has effects of warming kidney, tonifying yang, invigorating qi, relieving asthma, warming spleen and relieving diarrhea. According to the found in the literature, the compounds identified and contained in the fructus psoraleae are mainly coumarins, flavonoids, monoterpene phenols and the like. Psoralea fruit shows various biological activities. Wherein, the monoterpene phenols are the material basis for resisting tumor, inhibiting cell proliferation, resisting oxidation, resisting bacteria, resisting inflammation, resisting depression and protecting liver. Psoralen A (compound structure shown in figure 1) is used as one of main components in psoralen flavonoids, and has antitumor activity, ABTS free radical scavenging effect and strong alpha-glucosidase inhibiting effect. The invention discovers that psoralen has the function of protecting cochlear hair cell injury caused by various reasons for the first time.
Disclosure of the invention
The invention aims to provide a psoralen drug-loaded pellet and application thereof in preparing an auricular point paste for preventing noise hearing damage.
The technical scheme adopted by the invention is as follows:
the invention provides an application of psoralen in preparing a medicament for preventing or treating cochlear hair cell injury.
The medicine is an auricular point patch for preventing noise hearing impairment.
The auricular point patch bean is a psoralen hydrogel auricular point patch bean, wherein the psoralen hydrogel auricular point patch bean is obtained by sticking psoralen drug-loaded pellets on psoralen hydrogel and drying in the shade at room temperature for 24 hours; the psoralen A hydrogel is prepared by mixing the following raw materials in parts by mass: psoralen 0.002-0.005%, sodium polyacrylate (AH-105X) 4.0-7.0%, dihydroxyaluminium glycolate 0.1-0.25%, tartaric acid 0.1-0.3%, glycerol 7.2-18.0%, polyethylene glycol (PEG 400) 2.4-5.0%, sodium carboxymethylcellulose (CMC-NA) 0.1-1%, disodium ethylene diamine tetraacetate (EDTA-2NA) 0.1-1%, polyvinylpyrrolidone (PVP K90) 0.6-2.4%, azone 0.1-1.2%, and the balance of deionized water, the total amount is 100%; the psoralen A drug-loaded pellet is prepared by coating a carrier with a drug-loaded coating solution and then coating with a film coating solution; the carrier is a medical stone pellet; the medicine-carrying coating liquid is prepared by mixing the following raw materials in parts by mass: 0.02-0.05% of psoralen, 0.2-0.5% of adhesive and 70% of ethanol aqueous solution with volume concentration as solvent; the adhesive is hydroxypropyl methylcellulose (HPMC K15M) or sodium carboxymethylcellulose (CMC-Na); the film coating liquid is prepared by mixing the following raw materials in percentage by mass: 7.2% of Opadry, 1% of magnesium stearate and deionized water as a solvent.
Further, the psoralen hydrogel is prepared by mixing the following raw materials in parts by mass: 0.003-0.004% of psoralen, 5.0-7.0% of sodium polyacrylate (AH-105X), 0.1-0.2% of dihydroxyaluminium glycinate, 0.2-0.25% of tartaric acid, 7.2-12% of glycerol, 4.0-5.0% of polyethylene glycol (PEG 400), 0.2-0.4% of sodium carboxymethylcellulose (CMC-NA), 0.2-0.3% of disodium ethylenediamine tetraacetic acid (EDTA-2NA), 0.6-1.5% of polyvinylpyrrolidone (PVP K90), 0.1-1.2% of azone and deionized water as a solvent, wherein the total amount is 100%.
Furthermore, the psoralen hydrogel is prepared by mixing the following raw materials in parts by mass: the final concentration of psoralen is 0.002%, sodium polyacrylate (AH-105X) 5.4%, dihydroxyaluminium aminoacetate 0.19%, tartaric acid 0.23%, glycerin 15.77%, polyethylene glycol (PEG 400) 4.5%, sodium carboxymethylcellulose (CMC-NA) 0.33%, disodium ethylenediamine tetraacetate (EDTA-2NA) 0.25%, polyvinylpyrrolidone (PVP K90) 0.62%, azone 0.6%, and the balance of deionized water, and the total amount is 100%.
Further, the carrier is porous medical stone micro-pills (food safety level) with the diameter of 2 mm.
Further, the medicine-carrying coating liquid is prepared by mixing the following raw materials in parts by mass: psoralen 0.02-0.04%, hydroxypropyl methylcellulose (HPMC K15M) 0.2-0.4%, and 70% ethanol water solution; most preferably 0.03% of psoralen, 0.3% of hydroxypropyl methylcellulose (HPMC K15M) and 70% ethanol aqueous solution of solvent.
Further, the psoralen hydrogel is prepared by the following method: mixing sodium polyacrylate, sodium carboxymethylcellulose, disodium ethylene diamine tetraacetate and dihydroxyaluminum glycerate according to the formula amount, grinding uniformly by using a mortar, adding glycerol and polyethylene glycol, and stirring uniformly at a low speed to obtain an oil phase; dissolving tartaric acid in deionized water, adding polyvinylpyrrolidone, and stirring for 10min to obtain water phase; pouring all the water phase into all the oil phase, stirring for 15min, adding psoralen and azone, and mixing to obtain psoralen hydrogel.
Further, the psoralen drug-loaded pellet is prepared by the following method: (1) mixing psoralen, adhesive (preferably hydroxypropyl methylcellulose) and 70% ethanol water solution according to formula amount, ultrasonic dissolving at 50Hz for 15min, stirring at low speed for 45min, and mixing to obtain coating solution; coating the carrier with the drug-loaded coating solution by adopting a fluidized bed bottom spraying mode, and coating for 2 hours under the conditions of material temperature of 33-35 ℃, fan frequency of 10-15Hz, atomization pressure of 0.1-0.3kPa and peristaltic pump rotating speed of 1.0-2.0 ml/min (preferably material temperature of 35 ℃, fan frequency of 15Hz, atomization pressure of 0.2kPa and peristaltic pump rotating speed of 1.5ml/min) to obtain drug-loaded pellets; (2) dissolving the Opadry powder in deionized water according to the formula amount, adding magnesium stearate, and stirring at low speed for 45min to prepare a film coating solution; (3) coating the drug-loaded pellets prepared in the step (1) with a film coating solution in a fluidized bed bottom spraying manner, and coating for 2 hours under the conditions of the material temperature of 33-35 ℃, the fan frequency of 10-15Hz, the atomizing pressure of 0.1-0.3kPa and the peristaltic pump of 1.0-2.0 ml/min (the material temperature is preferably 35 ℃, the fan frequency of 15Hz, the atomizing pressure of 0.2kPa and the rotation speed of the peristaltic pump of 1.5ml/min) to obtain the psoralen drug-loaded pellets.
The auricular point patch bean is characterized in that psoralen drug-loaded pellets are fixed on an inner auricular point through psoralen hydrogel, and the patch is replaced every 12 hours; the patch is pressed to stimulate the inner ear acupoint 3-4 times per day by a gentle massage method; every seven days is a course of treatment.
The invention adopts skeleton pellets containing effective drugs to replace the traditional auricular point sticking media such as the seed of the cowherb, magnetite and the like; optimizing technical parameters of a fluidized bed, and preparing medical stone drug-loaded pellets with uniform quality; the traditional medical rubberized fabric layer is replaced by the medicine-containing hydrogel layer; the medical hydrogel with better transdermal drug release function is prepared by observing the transdermal absorption performance and optimizing the gel proportion; the method takes the cochlear hair cell oxidative damage resistance as a main efficacy evaluation mode, and preferably selects the traditional Chinese medicine monomer for preventing cochlear hair cell damage and the application concentration range thereof; finally, the product of the medicine-containing auricular point patch bean product which integrates the functions of pressing and stimulating auricular point and transdermal absorption of the medicine and has definite effect of preventing auricular injury is prepared.
Compared with the prior art, the invention has the following beneficial effects: the effective components of the psoralen hydrogel auricular point patch bean are derived from kidney-tonifying traditional Chinese medicine bone fat supplement, the psoralen hydrogel auricular point patch bean is prepared by combining with the traditional auricular point treatment means of the traditional Chinese medicine, and pharmacodynamics verifies that the preparation has a targeted protection effect on cochlear cell injury caused by various reasons, can press and stimulate auricular points, can stably and effectively absorb the medicine through skin, and jointly protects hearing.
(IV) description of the drawings
FIG. 1 shows the structure of psoralen.
Fig. 2 is a photograph of a carrier and a schematic diagram of an ear patch, wherein a is maifanite with a diameter of 2mm and a magnification of 16.9 × (zeiss stereomicroscope), B is a micropore on the surface of a maifanite sphere and a magnification of 163 × (zeiss stereomicroscope), C is a drug-loaded pellet, a magnification of 16.9 × (zeiss stereomicroscope), and D is a schematic diagram of the whole ear patch.
Fig. 3 is an overall schematic diagram (a) and a use schematic diagram (B) of psoralen hydrogel auricular point patch bean.
FIG. 4 is a graph showing MTT assay to determine the effect of psoralen on the survival rate of acetaminophen-induced damaged HEI-OC1 cells; note that P <0.05 and P <0.01 compared to the model group.
FIG. 5 is a flow apoptosis method for detecting the effect of psoralen on the apoptosis rate of acetaminophen-induced damage HEI-OC1 cells.
FIG. 6 is a flow cytometry assay to examine the effect of psoralen on ROS reactive oxygen species production in acetaminophen-injured mouse cochlear hair cells.
(V) detailed description of the preferred embodiments
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
example 1: evaluation of effect of psoralen on preventing and protecting cochlear hair cell injury
Studies have shown that high intensity noise exposure or ototoxic drug exposure can cause apoptosis, degeneration, necrosis and central auditory processing dysfunction of cochlear hair cells, while extracochlear hair cell death is the earliest and most prominent pathological change of noise-induced cochlear hair cell damage. Wherein the accumulation of Reactive Oxygen Species (ROS) is responsible for hearing impairment. Therefore, after the psoralen is subjected to prevention and protection, MTT cell activity detection, flow cell apoptosis detection and intracellular Reactive Oxygen Species (ROS) level detection are carried out on a cochlear hair cell (HEI-OC1) damage model after the stimulation of the acetaminophen by the ototoxic drug, so that the ear damage prevention and protection effect of the psoralen is verified.
1. Materials:
preparing psoralen mother liquor: 0.0050g of psoralen A is dissolved in 500 μ l of dimethyl sulfoxide (DMSO) to obtain 20mM mother solution of psoralen A.
Preparation of acetaminophen mother liquor: acetaminophen (0.0378 g) was dissolved in 630. mu.l DMSO to prepare a 20mM stock solution of acetaminophen.
MTT staining solution, 5 mg/mL; DMEM medium (Gibco); PBS (pH 7.2-7.4); pancreatin. FBS fetal bovine serum (hangzhou sijiqing); FITC Annexin V Apoptosis Detection Kit I (BD) Kit; active oxygen detection kit (Biyuntian).
2. Experimental cell lines: mouse cochlear hair cells (HEI-OC1) were purchased from Biofeng, Inc. (original strain purchased from House Ear Institute, Inc.; CVCL _ D899).
3. Cell line culture conditions: 33 ℃ and 10% CO2。
4. The experimental method comprises the following steps:
taking HEI-OC1 cells in logarithmic growth phase, and adjusting cell density to 2 × 104The cells were then seeded in 96-well plates. Adding 100 μ L of high-glucose DMEM complete medium into each well, and standing at 33 deg.C and 10% CO2Culturing in a constant temperature incubator for 24 h. After the cells adhere to the wall, the cells are replaced by DMEM high-glucose culture solution containing FBS with the volume concentration of 2%, after the cells are continuously cultured for 24 hours, the cells are replaced by DMEM high-glucose culture solution (100 mu L/hole) containing FBS with the volume concentration of 5%, the 96-well plate is provided with zero adjusting holes and is divided into a blank group, an experimental group and a control group (namely an injury model group), wherein the experimental group comprises a psoralen high-concentration group, a psoralen medium-concentration group and a psoralen low-concentration group, and each group is provided with 3 parallel holes.
The experimental groups were added with psoralen mother liquor to final concentrations of 100nM (high), 10nM (medium) and 1nM (low), respectively. After further incubation for 24h, DMEM high-glucose medium (100. mu.L/well) containing acetaminophen at a final concentration of 400. mu.M and 5% FBS by volume was added to the experimental and control groups, respectively, and drug-free DMEM high-glucose medium containing DMSO at a volume concentration of 0.1% was added to the blank group and the incubation was continued in the incubator for 24 h.
5. Effect of psoralen on survival of Acetaminophen-injured mouse cochlear hair cells following intervention
And 4, adding 20 mu L of freshly prepared MTT solution (5mg/mL) into each hole, continuously culturing for 4h, absorbing the supernatant in the plate, adding 150 mu L of dimethyl sulfoxide (DMSO) into each hole, placing the plate on a shaker, vibrating the plate at a low speed for 10min (60 times/min), and measuring the absorbance (A) value at 570nm by using an enzyme-linked immunosorbent calibration instrument. The cell survival rate of each group was calculated from the absorbance, and three duplicate wells were set for each group.
Cell viability (%) × (experimental well a value-blank well a value)/(control well a value-blank well a value) ] × 100%
The MTT method is adopted to primarily detect and determine that psoralen has protective effect on acetaminophen-induced damaged HEI-OC1 cells, and the figure is shown in figure 4. The experimental result shows that in the model of acetaminophen (400 mu M, 24h) induced injury, the cell survival rate of the acetaminophen injury model group (0nM psoralen A) is (87.854 +/-5.2178)%; under the condition that the concentration of psoralen is 100, 10 and 1nM (high, medium and low dose groups), the cell survival rates are respectively (94.558 +/-4.253)%, (103.309 +/-6.149)%, and (106.922 +/-4.350)%, compared with the injury model group, the difference of the low dose group has statistical significance (P <0.05), and the difference of the medium dose group has significant difference (P < 0.001). Therefore, 1-100nM psoralen has protective effect on acetaminophen-induced HEI-OC1 cell injury.
6. Effect of psoralen on apoptosis rate of Acetaminophen-injured mouse cochlear hair cells after intervention
And 4, removing the culture solution, washing with PBS once, adding 300 mu l of pancreatin into each hole for digestion, treating the cells according to the instruction shown by a FITCANNEXIN V Apoptosis Detection Kit I (BD) Kit, carrying out double-color marking on FITC and PI, and establishing a single-staining group and a blank non-staining group. After incubation and staining for 15min in the absence of light on ice, each group of HEI-OC1 was detected by flow cytometry for apoptosis.
And (3) data analysis: annexin V FITC/PI flow cytometry distinguishes normal, necrotic and apoptotic cells in an experimental sample. FITC and PI fluorescence were used as a two-parameter dot plot, and the cells were divided into 4 zones, zone 1: mechano-damaged cells (Annexin VFITC-/PI +); zone 2: late apoptotic or necrotic cells (Annexin V FITC +/PI +); zone 3: viable cells (Annexin VFITC-/PI-); zone 4: early apoptotic cells (Annexin V FITC +/PI-). The proportion of 4 groups of cells was counted and compared with the injury model group at different concentrations of psoralen, respectively, as shown in fig. 5.
The experimental results are as follows: the viable cell rates of the psoralen low-dose group (1nM) and the medium-dose group (10nM) were 76.21% and 78.33%, respectively, which were higher and less apoptotic than those of the injury model group compared to 70.10%. The result shows that psoralen has protective effect on acetaminophen-induced damaged HEI-OC1 cells.
7. Effect of psoralen on ROS reactive oxygen species production by Acetaminophen-injured mouse cochlear hair cells following psoralen intervention
The experimental principle is as follows: the active oxygen detection kit is a kit for detecting active oxygen by using a fluorescent probe DCFH-DA. DCFH-DA has no fluorescence, can freely pass through cell membranes, and can be hydrolyzed by intracellular esterase to generate DCFH after entering cells. DCFH, however, does not permeate the cell membrane, thus allowing the probe to be easily loaded into the cell. Intracellular reactive oxygen species can oxidize non-fluorescent DCFH to produce fluorescent DCF (detectable by the FITC channel). The level of reactive oxygen species in the cell can be known by measuring the fluorescence of DCF.
Step 4, collecting each group of cells, diluting DCFH-DA with serum-free DMEM culture solution according to a volume ratio of 1:1000 to a final concentration of 10. mu.M, collecting the cells, suspending the cells in the diluted DCFH-DA to a cell concentration of 1 × 106mL, incubation in a 37 ℃ cell incubator for 20 min. Mix by inversion every 5min to bring the probe and cells into intimate contact. The cells were washed three times with serum-free DMEM cell culture medium to sufficiently remove DCFH-DA that had not entered the cells. The flow cytometer FITC channel detected the ROS production levels in each group of HEI-OC1 cells, and the results are shown in FIG. 6.
And (3) data analysis: the cells were divided into negative and positive cell populations by using a control group (see fig. 6) without a fluorescent probe, and the level of ROS production was determined to be different by the difference in the ratio between the negative cell population and the positive cell population (the stronger the fluorescence intensity, the higher the ratio of the positive cell population, i.e., the higher the ROS production level).
The experimental results are as follows: the proportion of blank group positive cell population and model group positive cell population is 5.98 percent and 27.92 percent respectively, namely the ROS level in model group HEI-OC1 cells after acetaminophen damage is obviously increased. Under the prevention condition of the psoralen high, medium and low dose groups, the proportion of positive cell populations is 24.83%, 18.72% and 19.54%, the results are all smaller than the positive proportion of model group cells, and the proportion of the positive cells in the medium dose group is the lowest. The result can show that the psoralen A reduces the ROS (reactive oxygen species) generation level of mouse cochlear hair cells damaged by acetaminophen after the psoralen A is dried, namely the psoralen A is proved to be capable of effectively reducing the accumulation of ROS in the cochlear hair cells and reducing the damage of cells.
Example 2: preparation of psoralen A drug-loaded pellet
1. Material
The carrier is a medical stone pellet with the diameter of about 2mm, is a scientific name quartz dilonge (purchased from Hebei Hongyao mineral company), is a natural silicate mineral, is nontoxic and harmless to organisms, has large sphere hardness and micropores (shown as A and B in figure 2) on the surface, has strong adsorption effect, and is an ideal framework material of the drug-carrying pellet; opadry (available from shanghai kalekang coating technology limited) is a common drug coating material, can optimize appearance, improve odor, effectively prevent light and moisture, and increase drug stability.
2. The instrument comprises the following steps: electronic balance, precision force-increasing electric stirrer, 300ml beaker, fluidized bed (Glatt, germany).
3. Preparation of psoralen A drug-loaded pellet
(1) Fluidized bed liquid lamination method for loading medicine
The psoralen coating solution has the formula 1: psoralen A0.03%, hydroxypropyl methylcellulose (HPMC K15m) 0.3%, and solvent 70% ethanol water solution.
Psoralen coating solution formula 2: psoralen A0.03% and solvent 70% ethanol water solution.
Psoralen coating solution formula 3: psoralen A0.03%, sodium carboxymethylcellulose (CMC-Na) 0.3%, and solvent 70% ethanol water solution.
Respectively preparing a formula 1 according to the formula amount: 0.03g psoralen, 0.3g HPMC K15m, and 99.67g ethanol water solution with volume concentration of 70%.
And (2) formula: 0.03g of psoralen and 99.97g of 70% ethanol aqueous solution with volume concentration.
And (3) formula: 0.03g of psoralen, 0.3g of CMC-Na and 99.67g of 70% ethanol aqueous solution with volume concentration. Dissolving the above materials in 50Hz ultrasonic wave for 15min, stirring at low speed (preferably without foaming) for 45min, and mixing to obtain 100g of coating solution.
Taking 50 medical stones in 3 groups, wherein the total weight of the group 1 is 0.6225g, the total weight of the group 2 is 0.6345g, and the total weight of the group 2 is 0.610g, and the average particle weight of each group is shown in Table 1. The medical stones of groups 1 to 3 are respectively placed in a fluidized bed coating machine, 100g of drug-loaded coating liquid of the formula 1, the formula 2 and the formula 3 with the same number is used for coating a carrier in a fluidized bed bottom spraying mode, the carrier is coated for 2 hours under the conditions that the material temperature is 35 ℃, the fan frequency is 15Hz, the atomizing pressure is 0.2kPa and the rotating speed of a peristaltic pump is 1.5ml/min, the drug-loaded medical stones are obtained and are respectively marked as drug-loaded medical stone 1, drug-loaded medical stone 2 and drug-loaded medical stone 3, and the average drug-loaded amount of the medical stones before and after drug loading is calculated according to the formula (1) and.
Formula (1): the average drug loading is (total weight after drug loading-total weight before drug loading) × "total drug amount/(total weight of adjuvant + total drug) ]/50, wherein the adjuvant refers to adhesive, and the drug refers to psoralen.
As can be seen from Table 1: the average grain weight increment of the medical stones in each group is respectively 0.45mg, 0.05mg and 0.3mg, so that the drug-loading rates of different auxiliary materials are as follows: HPMC K15m > CMC-Na > without auxiliary material. Therefore, the formula 1 is the optimal choice, namely the formula of the final drug-loaded coating solution is 0.03 percent of psoralen, 0.3 percent of hydroxypropyl methylcellulose (HPMC K15m) and the solvent is ethanol water solution with the volume concentration of 70 percent. Each Maifanitum granule can carry 40.9 μ g psoralen, and the Maifanitum after carrying the medicine is shown in figure 2C.
TABLE 1 weight change and drug loading of Maifanitum before and after drug loading
(2) Film coating
The film coating liquid comprises the following components in parts by mass: 7.2% of Opadry, 1% of magnesium stearate and the balance of deionized water, wherein the total amount is 100%.
According to the formula amount, 7.2 g of Opadry powder is dissolved in 91.8g of deionized water, 1g of magnesium stearate is added, and the mixture is placed on a stirrer to be stirred at a low speed for 45min to prepare 100g of film coating liquid.
Placing 0.645g of the medicine-carrying medical stone 1 prepared in the step (1) in a fluidized bed coating machine, coating 100g of film coating liquid on the medicine-carrying medical stone in a fluidized bed bottom spraying mode, and coating for 2 hours under the conditions of material temperature of 35 ℃, fan frequency of 15Hz, atomizing pressure of 0.2kPa and peristaltic pump of 2.0ml/min to obtain 0.6644g of psoralen A medicine-carrying pellets, wherein the weight is increased by about 3 percent, and the particle diameter is not obviously changed.
Example 3: preparation of psoralen hydrogel
1. The psoralen A hydrogel comprises the following components in parts by mass:
the psoralen A hydrogel comprises the following components in parts by mass: the final concentration of psoralen is 0.002%, sodium polyacrylate (AH-105X) 5.4%, dihydroxyaluminium aminoacetate 0.19%, tartaric acid 0.23%, glycerin 15.77%, polyethylene glycol (PEG 400) 4.5%, sodium carboxymethylcellulose (CMC-NA) 0.33%, disodium ethylenediamine tetraacetate (EDTA-2NA) 0.25%, polyvinylpyrrolidone (PVPK90) 0.62%, azone 0.6%, and the balance of deionized water, and the total amount is 100%.
2. The preparation process comprises the following steps:
(1) the instrument comprises the following steps: an electronic balance, a precision force-increasing electric stirrer, a 300ml beaker, a spatula and a mortar.
(2) The preparation method comprises the following steps: mixing 5.4g of sodium polyacrylate, 0.33g of CMC-NA, 0.25g of EDTA-2NA and 0.19g of dihydroxyaluminum glycinate according to the formula amount, grinding uniformly by using a mortar, adding 15.77g of glycerol and 4.5g of PEG400, and uniformly stirring at a low speed (preferably without foaming) to obtain an oil phase; dissolving 0.23g tartaric acid in 72.108ml deionized water (pH 2.38), adding 0.62g PVP, and stirring for 10min to obtain water phase; pouring all water phase into all oil phase, stirring for 15min, adding 0.002g psoralen and 0.6g azone, and mixing to obtain 100g psoralen hydrogel (pH 6-7) with diameter of 8mm × 8 mm.
Example 4 preparation of psoralen hydrogel auricular point patch bean
Taking 100g of psoralen hydrogel prepared by the method in example 3 as an adhesive layer, drying in the shade at room temperature for 24h, covering a non-woven mucous membrane, and covering the psoralen drug-loaded pellet prepared in example 2 with the psoralen hydrogel to obtain the psoralen hydrogel auricular point patch bean, with reference to a in fig. 3A.
Claims (10)
1. An application of psoralen in preparing medicine for preventing and treating cochlear hair cell injury is disclosed.
2. The use of claim 1, wherein the medicament is an auricular point patch for the prevention of noise-induced hearing impairment.
3. The use of claim 2, wherein the auricular point patch is a psoralen hydrogel auricular point patch obtained by pasting a psoralen drug-loaded pellet on psoralen hydrogel and drying in the shade at room temperature;
the psoralen A hydrogel is prepared by mixing the following raw materials in parts by mass: 0.002-0.005% of psoralen, 4.0-7.0% of sodium polyacrylate, 0.1-0.25% of dihydroxyaluminium glycolate, 0.1-0.3% of tartaric acid, 7.2-18.0% of glycerol, 2.4-5.0% of polyethylene glycol, 0.1-1% of sodium carboxymethylcellulose, 0.1-1% of disodium ethylene diamine tetraacetate, 0.6-2.4% of polyvinylpyrrolidone, 0.1-1.2% of azone and the balance of deionized water, wherein the total amount is 100%;
the psoralen A drug-loaded pellet is prepared by coating a carrier with a drug-loaded coating solution and then coating with a film coating solution; the carrier is a medical stone pellet; the medicine-carrying coating liquid is prepared by mixing the following raw materials in parts by mass: 0.02-0.05% of psoralen, 0.2-0.5% of adhesive and 70% of ethanol aqueous solution with volume concentration as solvent; the adhesive is hydroxypropyl methyl cellulose or sodium carboxymethyl cellulose; the film coating liquid is prepared by mixing the following raw materials in percentage by mass: 7.2% of Opadry, 1% of magnesium stearate and deionized water as a solvent.
4. The application of claim 2, wherein the psoralen hydrogel is prepared by mixing the following raw materials in parts by mass: 0.003-0.004% of psoralen, 5.0-7.0% of sodium polyacrylate, 0.1-0.2% of dihydroxyaluminium glycolate, 0.2-0.25% of tartaric acid, 7.2-12% of glycerol, 4.0-5.0% of polyethylene glycol, 0.2-0.4% of sodium carboxymethylcellulose, 0.2-0.3% of disodium ethylenediamine tetraacetic acid, 0.6-1.5% of polyvinylpyrrolidone, 0.1-1.2% of azone and the balance of deionized water, wherein the total amount is 100%.
5. The application of claim 2, wherein the psoralen hydrogel is prepared by mixing the following raw materials in parts by mass: the final concentration of psoralen is 0.002%, sodium polyacrylate 5.4%, dihydroxyaluminium aminoacetate 0.19%, tartaric acid 0.23%, glycerin 15.77%, polyethylene glycol 4.5%, sodium carboxymethylcellulose 0.33%, disodium ethylenediamine tetraacetate 0.25%, polyvinylpyrrolidone 0.62%, azone 0.6%, and the balance of deionized water, and the total amount is 100%.
6. The use of claim 2, wherein said carrier is a porous Maifanitum pellet having a diameter of 2 mm.
7. The application of claim 2, wherein the drug-loaded coating solution is prepared by mixing the following raw materials in parts by mass: psoralen 0.02-0.04%, hydroxypropyl methylcellulose 0.2-0.4%, and 70% ethanol water solution as solvent.
8. The application of claim 2, wherein the drug-loaded coating solution is prepared by mixing the following raw materials in parts by mass: psoralen A0.03%, hydroxypropyl methylcellulose 0.3%, and 70% ethanol water solution.
9. The use of claim 2, wherein said psoralen-based hydrogel is prepared by the following method: mixing sodium polyacrylate, sodium carboxymethylcellulose, disodium ethylene diamine tetraacetate and dihydroxyaluminum glycerate according to the formula amount, grinding uniformly by using a mortar, adding glycerol and polyethylene glycol, and stirring uniformly at a low speed to obtain an oil phase; dissolving tartaric acid in deionized water, adding polyvinylpyrrolidone, and stirring for 10min to obtain water phase; pouring all the water phase into all the oil phase, stirring for 15min, adding psoralen and azone, and mixing to obtain psoralen hydrogel.
10. The use of claim 2, wherein the psoralen A drug-loaded pellet is prepared by the following method: (1) mixing psoralen and adhesive with 70% ethanol water solution, ultrasonic dissolving at 50Hz for 15min, stirring at low speed for 45min, and making into coating solution; coating the carrier with the drug-loaded coating solution by adopting a fluidized bed bottom spraying mode, and coating for 2 hours under the conditions of material temperature of 33-35 ℃, fan frequency of 10-15Hz, atomization pressure of 0.1-0.3kPa and peristaltic pump rotating speed of 1.0-2.0 ml/min to obtain drug-loaded pellets; (2) dissolving the Opadry powder in deionized water according to the formula amount, adding magnesium stearate, and stirring at low speed for 45min to prepare a film coating solution; (3) coating the drug-loaded pellets prepared in the step (1) with a film coating solution in a fluidized bed bottom spraying manner, and coating for 2 hours under the conditions of material temperature of 33-35 ℃, fan frequency of 10-15Hz, atomization pressure of 0.1-0.3kPa and peristaltic pump of 1.0-2.0 ml/min to obtain the psoralen drug-loaded pellets.
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