CN113679657B - Silk fibroin microneedle transdermal patch for treating insomnia and preparation method thereof - Google Patents
Silk fibroin microneedle transdermal patch for treating insomnia and preparation method thereof Download PDFInfo
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- CN113679657B CN113679657B CN202110871605.8A CN202110871605A CN113679657B CN 113679657 B CN113679657 B CN 113679657B CN 202110871605 A CN202110871605 A CN 202110871605A CN 113679657 B CN113679657 B CN 113679657B
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
- A61K31/404—Indoles, e.g. pindolol
- A61K31/4045—Indole-alkylamines; Amides thereof, e.g. serotonin, melatonin
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- 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
- A61K47/40—Cyclodextrins; Derivatives thereof
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- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
- A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/20—Hypnotics; Sedatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
- A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
- A61M2037/0046—Solid microneedles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
- A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
- A61M2037/0053—Methods for producing microneedles
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a silk fibroin microneedle transdermal patch for treating insomnia and a preparation method thereof, wherein the preparation method comprises the following steps: respectively preparing aqueous solutions of a microneedle matrix modification additive, a drug dissolution additive and a drug release additive; adding melatonin into the drug dissolving additive aqueous solution to obtain a melatonin solution; adding the microneedle matrix modification additive aqueous solution and the melatonin solution into the silk fibroin aqueous solution, and adding the drug release additive aqueous solution to obtain a mixed solution II; and pouring the mixed solution II into a microneedle mould, and carrying out vacuumizing, defoaming, drying and demoulding to obtain the silk fibroin microneedle transdermal patch for treating insomnia. The melatonin microneedle patch prepared by the invention can quickly reach a certain blood level and keep the higher concentration of the medicine in the body for a long time; meanwhile, the microneedle transdermal patch avoids the first pass effect of the liver and can improve the bioavailability of the melatonin.
Description
Technical Field
The invention belongs to the technical field of transdermal drug delivery, and particularly relates to a silk fibroin microneedle transdermal patch for treating insomnia and a preparation method thereof.
Background
Melatonin, originally found in the bovine pineal gland in 1958, is a hormone synthesized by the pineal gland and released at night, and exogenous melatonin administration has been shown to have phase change or retiming biological clock rhythm and sleep-wake schedule effects. The physiological role of melatonin in regulating seasonal and circadian rhythms is thought to be mediated by specific, high affinity and G-protein coupled cell membrane receptors. Currently, the oral route is the most studied. Oral melatonin enters the liver through the digestive tract, so that the first pass effect of the liver is caused, and the bioavailability of the medicine is reduced by only about 20%. Meanwhile, oral melatonin is difficult to reach a certain blood concentration and maintain a high concentration for a long time. Therefore, transdermal delivery of melatonin may be a good route of administration.
The micro-needle transdermal drug delivery system is a novel mode of the transdermal drug delivery system, and is a needle-shaped complex structure with micron-sized dimensions. Compared with other transdermal drug delivery methods, the microneedle has unique advantages that: (1) compared with chemical permeation promotion and ion introduction, the microneedle can puncture the stratum corneum to generate tiny pore passages for the entrance of macromolecular drugs; (2) the micro-needle punctures the channel of the human body by only micron level, has no damage to the skin and is not enough to touch the nerve endings, so the pain sensation is almost not generated; (3) the micro-needle is combined with other methods, so that the transdermal penetration capacity of the medicament can be further improved.
In the prior art, a chinese patent with application number CN201811017105.2 discloses a preparation method of a melatonin microneedle patch, which includes the following steps: pouring a melatonin-containing polymer material aqueous solution on a mold with a micropore matrix, filling the polymer material aqueous solution into micropores on the mold, attaching a prefabricated backing layer to the mold, attaching the backing layer to the back of melatonin microneedles formed in the micropores, freezing, thawing, uncovering and drying to obtain the melatonin microneedle patch. But the release speed is slow, and the elimination speed of the medicine in the body is fast.
Disclosure of Invention
The invention aims to provide a silk fibroin microneedle transdermal patch for treating insomnia and a preparation method thereof, wherein melatonin is delivered transdermally, a certain blood level can be rapidly achieved, and a higher concentration of a medicine in a body can be maintained for a long time.
According to the technical scheme of the invention, the preparation method of the silk fibroin microneedle transdermal patch for treating insomnia comprises the following steps,
s1: respectively preparing a microneedle matrix modification additive aqueous solution, a drug dissolution additive aqueous solution and a drug release additive aqueous solution;
s2: adding melatonin into the drug dissolving additive aqueous solution, and uniformly mixing to obtain a melatonin solution;
s3: adding the microneedle matrix modified additive aqueous solution obtained in the step S1 and the melatonin solution obtained in the step S2 into the silk fibroin aqueous solution with the concentration of 20-60mg/mL to obtain a mixed solution I;
s4: adding the aqueous solution of the drug release additive obtained in the step S1 into the mixed solution I to obtain a mixed solution II, wherein the concentration of melatonin in the mixed solution II is 0.5-20mg/mL, the concentration of the microneedle matrix modification additive is 4-25mg/mL, and the concentration of the drug release additive is 0.05-10mg/mL;
s5: and pouring the mixed solution II into a microneedle mould, vacuumizing, defoaming, drying and demoulding to obtain the silk fibroin microneedle transdermal patch for treating insomnia.
Further, the microneedle matrix modification additive is one or more of proline, sorbitol and xylitol.
Further, the concentration of the microneedle matrix modification additive aqueous solution is 100-1000mg/mL, and the preferred concentration is 300-900mg/mL.
Further, in the mixed solution II, the mass ratio of the microneedle matrix modification additive to the silk fibroin is 1-5:10.
further, the drug dissolution additive is one or more of isopropanol, n-propanol, n-butanol, isobutanol and 2-butanol.
Further, the concentration of the drug dissolution additive aqueous solution is 100-800mg/mL, preferably 300-800mg/mL.
Further, in the mixed solution II, the mass ratio of the drug dissolution additive to the silk fibroin is 1:5-20.
Further, the drug release additive is one or more of alpha-cyclodextrin, hydroxypropyl-beta-cyclodextrin and gamma-cyclodextrin.
Further, the concentration of the aqueous solution of the drug release additive is 10-100mg/mL.
Furthermore, the concentration of the melatonin in the melatonin solution is 5-60mg/mL, and the preferable concentration is 15-55mg/mL.
Further, the melatonin solution and the mixed solution II are stored at the temperature of 2-8 ℃ in the dark.
Further, the silk fibroin aqueous solution is obtained by degumming, dissolving, dialyzing and purifying silkworm silk.
Further, the concentration of the silk fibroin aqueous solution is 30-50mg/mL.
Preferably, the concentration of the melatonin in the mixed solution II is 5-15mg/mL, the concentration of the microneedle matrix modification additive is 10-25mg/mL, and the concentration of the drug release additive is 2-10mg/mL.
Furthermore, the concave hole of the microneedle mould is conical, the length is 200-800 μm, and the diameter of the bottom is 100-400 μm.
Furthermore, the microneedle mould is made of silicon rubber.
Further, in step S5, drying is carried out for 4-8h under the condition of constant temperature and humidity (20-30 ℃ and 55-65%) air circulation, and the drug dissolving additive is small molecular alcohol, so that the drug dissolving additive is completely volatilized and removed in the drying process.
On the other hand, the invention provides the silk fibroin microneedle transdermal patch for treating insomnia, which is prepared by the preparation method. The micro-needle body of the obtained fibroin protein micro-needle transdermal patch is in a solid conical shape, the length of the needle body is 200-800 mu m, the diameter of the bottom is 100-400 mu m, and the interval between the adjacent needle bodies is 0.2-2mm.
The invention principle is as follows:
the silk fibroin is a natural protein, has good mechanical property and plasticity, good compatibility with human tissues, biodegradability, no toxicity and easy processing and preparation. A large number of hydrophilic groups contained in the gel can be better included with water-soluble drugs, so that the lipid-rich stratum corneum is broken through, and the transdermal penetration capability of the drugs is improved.
Melatonin is a fat-soluble drug, is insoluble in water and is difficult to be uniformly mixed with a silk fibroin solution. The intervention of the drug dissolving additive changes the molecular stereo arrangement of unit cells in the original crystal structure of the melatonin, so that the melatonin enters the drug dissolving additive in a molecular state. And then can be uniformly blended with silk fibroin to form uniform microneedles. The melatonin is uniformly distributed in the micro-needle, and the uniform release speed is obtained in the subsequent transdermal drug release. The microneedle matrix modification additive is added to promote silk fibroin to form a certain crystal structure, reduce the dissolution and dissolution rate, form a certain swelling pore diameter and control the release speed of melatonin.
The drug release additive is added to accelerate the release speed of the melatonin and improve the cumulative release amount of the melatonin. Due to the high hydrophilicity of the drug release additive, the drug can be dissolved and released in vivo more easily, and higher blood concentration can be achieved in a short time. Meanwhile, the solubility of the melatonin in body fluid is improved, the bioavailability of the melatonin is further improved, and the drug effect is enhanced, so that the addition amount of the drug is reduced. Therefore, after the drug release additive and the drug are included, the proportion, the type and the existence form of the drug can influence the solubility of the drug in the inclusion compound. Meanwhile, for melatonin, 4 controlled release effects, namely immediate release, delayed release (time-controlled release), extended release and controlled release, can be achieved through the inclusion effect of cyclodextrin on melatonin. The release speed can be improved or regulated by selecting proper cyclodextrin and adding proper amount and reasonably designing the formula of the melatonin microneedle.
Compared with the prior art, the technical scheme of the invention has the following advantages: the melatonin microneedle patch prepared by the invention can quickly reach a certain blood level and keep the higher concentration of the in-vivo drug for a long time. Meanwhile, the micro-needle transdermal patch avoids the first-pass effect of the liver and can improve the bioavailability of the melatonin.
Drawings
Fig. 1 is a microphotograph of a microneedle patch prepared in example 1 of the present invention.
Fig. 2 is a plasma-drug concentration graph of the microneedle patch prepared in example 1 of the present invention in rats.
Fig. 3 is a graph showing the in vitro transdermal drug release profile of the microneedle patches prepared in examples 1 to 6 of the present invention.
Detailed Description
The present invention is further described below in conjunction with the drawings and the embodiments so that those skilled in the art can better understand the present invention and can carry out the present invention, but the embodiments are not to be construed as limiting the present invention.
Preparation example 1
Preparation of silk fibroin solution: 3g of NaHCO are weighed 3 And 1g of Na 2 CO 3 Dissolving in 4000mL deionized water, heating to boil, adding 80g silk, slightly boiling at 98 deg.C for 30min, taking out, and washing with deionized water. Repeating the steps for three times, and then placing the mixture in a 60 ℃ drying oven for drying to obtain the silk fibroin fiber. Preparing 9.3mol/L LiBr aqueous solution, heating 100mL LiBr aqueous solution to 65 ℃ in a water bath, adding 15g of silk fibroin fiber for many times, stirring and dissolving, and then continuing to heat and stir for 40min. The liquid is filled into a dialysis bag and placed into deionized water for dialysis for 72 hours, and the deionized water is replaced every 2 hours. Filtering the dialyzed solution to obtain a silk fibroin solution. And taking a small amount of solution, drying, weighing and calculating the mass fraction of the solution.
Example 1
Preparing a small molecule additive aqueous solution: weighing proline, adding the proline into deionized water, and uniformly mixing the proline and the deionized water by shaking to ensure that the concentration of the proline in the aqueous solution is 500mg/mL; weighing hydroxypropyl-beta-cyclodextrin, adding the hydroxypropyl-beta-cyclodextrin into deionized water, shaking and uniformly mixing to ensure that the concentration of the hydroxypropyl-beta-cyclodextrin in the aqueous solution is 100mg/mL; measuring isopropanol, adding the isopropanol into deionized water, and uniformly mixing the isopropanol and the deionized water by shaking to ensure that the concentration of the isopropanol in the aqueous solution is 600mg/mL.
Preparation of melatonin solution: weighing melatonin in a centrifuge tube, adding 1mL of isopropanol water solution, shaking and uniformly mixing to ensure that the concentration of the melatonin in the solution is 50mg/mL.
Preparation of mixed solution: slowly adding the proline aqueous solution and the melatonin solution into the silk fibroin aqueous solution while stirring, wherein the concentration of the silk fibroin aqueous solution is 50mg/mL; adding a hydroxypropyl-beta-cyclodextrin aqueous solution to obtain a mixed solution; the concentration of melatonin in the mixed solution is 15mg/mL, the concentration of proline is 10mg/mL, and the concentration of hydroxypropyl-beta-cyclodextrin is 5mg/mL, and the mixed solution is stored in a dark place at 4 ℃ after being fully mixed.
Injection molding of the solution: 3mL of the mixed solution is poured into a silicon rubber microneedle mould, and the silicon rubber microneedle mould is vacuumized for 3 times to remove the solution and bubbles in the microneedle mould plate, so that the solution fully enters the cavity of the needle body.
Drying of microneedle patches: and (3) placing the defoamed mould system in a constant-temperature constant-humidity room (25 ℃, 65%) in an air circulation place for drying for 8 hours, and demolding to obtain the melatonin-containing silk fibroin microneedle transdermal patch.
The microphotograph of the obtained silk fibroin microneedle transdermal patch is shown in fig. 1, and the length of a microneedle is 600 μm, and the diameter of the bottom surface is about 300 μm.
The plasma-drug concentration curve of the obtained silk fibroin microneedle transdermal patch used for rats is shown in fig. 2. As can be seen from the figure, the time for reaching the maximum blood concentration in the rat body is 0.41 plus or minus 0.03h, and the maximum blood concentration is 10.08 plus or minus 0.02ng/mL. The drug concentration in rats can maintain an effective drug concentration level and keep stable in a time range of 6 h. The microneedle patch prepared by the invention can reach a certain blood concentration in a short time, covers the whole sleep cycle and achieves the effect of assisting sleep.
Example 2
Preparing a small molecule additive aqueous solution: weighing sorbitol, adding the sorbitol into deionized water, shaking and uniformly mixing to ensure that the concentration of sorbitol in the aqueous solution is 400mg/mL; weighing hydroxypropyl-beta-cyclodextrin, adding the hydroxypropyl-beta-cyclodextrin into deionized water, shaking and uniformly mixing to ensure that the concentration of the hydroxypropyl-beta-cyclodextrin in the aqueous solution is 80mg/mL. Measuring n-propanol, adding into deionized water, shaking and uniformly mixing to ensure that the concentration of the n-propanol in the aqueous solution is 500mg/mL.
Preparation of melatonin solution: weighing melatonin in a centrifuge tube, adding 0.8mL of n-propanol aqueous solution, shaking and uniformly mixing to ensure that the concentration of the melatonin in the solution is 40mg/mL, and storing the melatonin solution at 4 ℃ in a dark place.
Preparation of mixed solution: slowly adding the sorbitol aqueous solution and the melatonin solution into the silk fibroin aqueous solution while stirring, wherein the concentration of the silk fibroin aqueous solution is 40mg/mL; adding a hydroxypropyl-beta-cyclodextrin aqueous solution to obtain a mixed solution; the concentration of melatonin in the mixed solution is 8mg/mL, the concentration of sorbitol is 10mg/mL, and the concentration of hydroxypropyl-beta-cyclodextrin is 5mg/mL, and the mixed solution is fully mixed and then placed at 4 ℃ for being stored in a dark place.
Injection molding of the solution: 3mL of the mixed solution is poured into a silicon rubber microneedle mould, and the silicon rubber microneedle mould is vacuumized for 3 times to remove the solution and bubbles in the microneedle mould plate, so that the solution fully enters the cavity of the needle body.
Drying of microneedle patches: and (3) drying the defoamed mould system for 8 hours at a constant temperature and humidity (25 ℃, 65%) air circulation position, and demolding to obtain the melatonin-containing silk fibroin microneedle transdermal patch.
Example 3
Preparing a small molecule additive aqueous solution: weighing sorbitol, adding the sorbitol into deionized water, shaking and uniformly mixing to enable the concentration of sorbitol in the aqueous solution to be 600mg/mL; weighing alpha-cyclodextrin, adding the alpha-cyclodextrin into deionized water, shaking and uniformly mixing to ensure that the concentration of the alpha-cyclodextrin in the aqueous solution is 50mg/mL. Measuring n-butyl alcohol, adding the n-butyl alcohol into deionized water, and shaking and mixing uniformly to ensure that the concentration of the n-butyl alcohol in the aqueous solution is 600mg/mL.
Preparation of melatonin solution: weighing melatonin in a centrifuge tube, adding 1mL of n-butanol aqueous solution, shaking and uniformly mixing to ensure that the concentration of the melatonin in the solution is 60mg/mL, and storing the melatonin solution at 4 ℃ in a dark place.
Preparation of mixed solution: slowly adding the sorbitol aqueous solution and the melatonin solution into the silk fibroin aqueous solution while stirring, wherein the concentration of the silk fibroin aqueous solution is 50mg/mL; adding a hydroxypropyl-beta-cyclodextrin aqueous solution to obtain a mixed solution; the concentration of melatonin in the mixed solution is 7mg/mL, the concentration of sorbitol is 20mg/mL, and the concentration of alpha-cyclodextrin is 5mg/mL, and the mixed solution is fully mixed and then placed at 4 ℃ for being stored in a dark place.
Injection molding of the solution: 3mL of the mixed solution is poured into a silicon rubber microneedle mould, and the silicon rubber microneedle mould is vacuumized for 3 times to remove the solution and bubbles in the microneedle mould plate, so that the solution fully enters the cavity of the needle body.
Drying of microneedle patches: and (3) placing the defoamed mould system in a constant-temperature constant-humidity room (25 ℃, 65%) in an air circulation place for drying for 8 hours, and demolding to obtain the melatonin-containing silk fibroin microneedle transdermal patch.
Example 4
Preparing a small molecule additive aqueous solution: weighing proline, adding the proline into deionized water, and uniformly mixing the proline and the deionized water by shaking to ensure that the concentration of the proline in the aqueous solution is 700mg/mL; weighing alpha-cyclodextrin, adding the alpha-cyclodextrin into deionized water, shaking and uniformly mixing to ensure that the concentration of the alpha-cyclodextrin in the aqueous solution is 30mg/mL. Measuring n-butanol, adding into deionized water, shaking and mixing uniformly to make the concentration of the n-butanol in the aqueous solution be 600mg/mL.
Preparation of melatonin solution: weighing melatonin in a centrifuge tube, adding 1.5mL of n-butyl alcohol aqueous solution, shaking and uniformly mixing to ensure that the concentration of the melatonin in the solution is 30mg/mL, and storing the melatonin solution at 4 ℃ in a dark place.
Preparation of mixed solution: slowly adding the proline aqueous solution and the melatonin solution into the silk fibroin aqueous solution while stirring, wherein the concentration of the silk fibroin aqueous solution is 40mg/mL; adding a hydroxypropyl-beta-cyclodextrin aqueous solution to obtain a mixed solution; the concentration of the melatonin in the mixed solution is 12mg/mL, the concentration of the proline is 15mg/mL, the concentration of the alpha-cyclodextrin is 3mg/mL, and the mixed solution is placed at 4 ℃ for dark storage after being fully mixed.
Injection molding of the solution: and 3mL of mixed solution is poured into a silicon rubber microneedle mould, and the silicon rubber microneedle mould is vacuumized for 3 times to remove the bubbles in the solution and the microneedle mould plate, so that the solution fully enters the cavity of the needle body.
Drying of microneedle patches: and (3) drying the defoamed mould system for 8 hours at a constant temperature and humidity (25 ℃, 65%) air circulation position, and demolding to obtain the melatonin-containing silk fibroin microneedle transdermal patch.
Example 5
Preparing a small molecule additive aqueous solution: weighing proline, adding the proline into deionized water, and uniformly mixing the proline and the deionized water by shaking to ensure that the concentration of the proline in the aqueous solution is 900mg/mL; weighing gamma-cyclodextrin, adding the gamma-cyclodextrin into deionized water, shaking and uniformly mixing to ensure that the concentration of the gamma-cyclodextrin in the aqueous solution is 70mg/mL. Measuring isobutanol, adding the isobutanol into deionized water, shaking and uniformly mixing to enable the concentration of the isobutanol in the water solution to be 800mg/mL.
Preparation of melatonin solution: weighing melatonin in a centrifuge tube, adding 2mL of isobutanol aqueous solution, shaking and uniformly mixing to ensure that the concentration of the melatonin in the solution is 55mg/mL, and storing the melatonin solution at 4 ℃ in a dark place.
Preparation of mixed solution: slowly adding the proline aqueous solution and the melatonin solution into the silk fibroin aqueous solution while stirring, wherein the concentration of the silk fibroin aqueous solution is 50mg/mL; adding a hydroxypropyl-beta-cyclodextrin aqueous solution to obtain a mixed solution; the concentration of melatonin in the mixed solution is 15mg/mL, the concentration of proline is 25mg/mL, and the concentration of gamma-cyclodextrin is 10mg/mL, and the mixed solution is stored in a dark place at 4 ℃ after being fully mixed.
Injection molding of the solution: 3mL of the mixed solution is poured into a silicon rubber microneedle mould, and the silicon rubber microneedle mould is vacuumized for 3 times to remove the solution and bubbles in the microneedle mould plate, so that the solution fully enters the cavity of the needle body.
Drying of microneedle patches: and (3) placing the defoamed mould system in a constant-temperature constant-humidity room (25 ℃, 65%) in an air circulation place for drying for 8 hours, and demolding to obtain the melatonin-containing silk fibroin microneedle transdermal patch.
Example 6
Preparing a small molecule additive aqueous solution: weighing xylitol, adding the xylitol into deionized water, and shaking and uniformly mixing to ensure that the concentration of the xylitol in the aqueous solution is 300mg/mL; weighing gamma-cyclodextrin, adding the gamma-cyclodextrin into deionized water, shaking and uniformly mixing to ensure that the concentration of the gamma-cyclodextrin in the aqueous solution is 10mg/mL. Weighing 2-butanol, adding into deionized water, shaking and mixing uniformly to ensure that the concentration of the 2-butanol in the aqueous solution is 300mg/mL.
Preparation of melatonin solution: weighing melatonin into a centrifuge tube, adding 1.5mL of 2-butanol aqueous solution, shaking and uniformly mixing to ensure that the concentration of the melatonin in the solution is 15mg/mL, and storing the melatonin solution at 4 ℃ in a dark place.
Preparation of mixed solution: slowly adding the xylitol aqueous solution and the melatonin solution into the silk fibroin aqueous solution while stirring, wherein the concentration of the silk fibroin aqueous solution is 50mg/mL; adding a hydroxypropyl-beta-cyclodextrin aqueous solution to obtain a mixed solution; the concentration of melatonin in the mixed solution is 5mg/mL, the concentration of xylitol is 15mg/mL, and the concentration of gamma-cyclodextrin is 2mg/mL, and the mixed solution is fully mixed and then placed at 4 ℃ for storage in a dark place.
Injection molding of the solution: and 3mL of mixed solution is poured into a silicon rubber microneedle mould, and the silicon rubber microneedle mould is vacuumized for 3 times to remove the bubbles in the solution and the microneedle mould plate, so that the solution fully enters the cavity of the needle body.
Drying of microneedle patches: and (3) drying the defoamed mould system for 8 hours at a constant temperature and humidity (25 ℃, 65%) air circulation position, and demolding to obtain the melatonin-containing silk fibroin microneedle transdermal patch.
Detection example 1
The graph of the in vitro transdermal drug release curve of the microneedle patch prepared in examples 1-6 is shown in fig. 3, and it can be seen that silk fibroin microneedles with different swelling degrees can be obtained by adding microneedle matrix modification additives with different kinds and proportions, so that the drug release speed in the microneedles and the time for balancing the drug release can be adjusted. The drug dissolution additive can enhance the dissolution amount of the drug, thereby obtaining microneedles with different drug loading amounts. The drug release additive can promote the release amount of the drug in the microneedle, and improve the accumulative release rate of the microneedle and the bioavailability of the melatonin.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications of the invention may be made without departing from the spirit or scope of the invention.
Claims (9)
1. A preparation method of a silk fibroin microneedle transdermal patch for treating insomnia is characterized by comprising the following steps,
s1: respectively preparing a microneedle matrix modification additive aqueous solution, a drug dissolution additive aqueous solution and a drug release additive aqueous solution;
the microneedle matrix modification additive is one or more of proline, sorbitol and xylitol; the medicine dissolving additive is one or more of isopropanol, n-propanol, n-butanol, isobutanol and 2-butanol; the drug release additive is one or more of alpha-cyclodextrin, hydroxypropyl-beta-cyclodextrin and gamma-cyclodextrin;
s2: adding melatonin into the drug dissolving additive aqueous solution, and uniformly mixing to obtain a melatonin solution;
s3: adding the microneedle matrix modified additive aqueous solution obtained in the step S1 and the melatonin solution obtained in the step S2 into the silk fibroin aqueous solution with the concentration of 20-60mg/mL to obtain a mixed solution I;
s4: adding the aqueous solution of the drug release additive obtained in the step S1 into the mixed solution I to obtain a mixed solution II, wherein the concentration of melatonin in the mixed solution II is 0.5-20mg/mL, the concentration of the microneedle matrix modification additive is 4-25mg/mL, and the concentration of the drug release additive is 0.05-10mg/mL;
s5: and pouring the mixed solution II into a microneedle mould, and carrying out vacuumizing, defoaming, drying and demoulding to obtain the silk fibroin microneedle transdermal patch for treating insomnia.
2. The method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1, wherein the concentration of the microneedle matrix modification additive aqueous solution is 100-1000 mg/mL.
3. The method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1 or 2, wherein in the mixed solution II, the mass ratio of the microneedle matrix modification additive to the silk fibroin is 1-5:10.
4. the method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1, wherein the concentration of the drug dissolution additive aqueous solution is 100-800 mg/mL.
5. The method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1, wherein the concentration of the drug release additive aqueous solution is 10-100mg/mL.
6. The method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1, wherein in the mixed solution II, the mass ratio of the drug release additive to the silk fibroin is 1:5-20.
7. The method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1, wherein the concentration of melatonin in the melatonin solution is 5-60 mg/mL.
8. The method for preparing the silk fibroin microneedle transdermal patch for treating insomnia as claimed in claim 1, wherein the concave hole of the microneedle mould is conical, the length is 200-800 μm, and the bottom diameter is 100-400 μm.
9. The silk fibroin microneedle transdermal patch for treating insomnia prepared by the preparation method of any one of claims 1-8.
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| CN202110871605.8A CN113679657B (en) | 2021-07-30 | 2021-07-30 | Silk fibroin microneedle transdermal patch for treating insomnia and preparation method thereof |
| PCT/CN2021/117078 WO2023004941A1 (en) | 2021-07-30 | 2021-09-08 | Silk fibroin microneedle transdermal patch for treating insomnia and preparation method therefor |
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| TW201330849A (en) * | 2012-01-20 | 2013-08-01 | Renascence Therapeutics Ltd | Pharmaceutical compositions for intranasal administration of melatonin |
| JP2016030072A (en) * | 2014-07-29 | 2016-03-07 | 富士フイルム株式会社 | Microneedle array |
| JP2017514646A (en) * | 2015-04-03 | 2017-06-08 | ソーチョウ ユニバーシティー | Swelled silk fibroin microneedle drug delivery system and method for producing the same |
| CN105833424A (en) * | 2016-03-21 | 2016-08-10 | 南通纺织丝绸产业技术研究院 | Silk fibroin micro-needle patch and preparation method thereof |
| CN107233297A (en) * | 2017-06-16 | 2017-10-10 | 广州新济薇娜生物科技有限公司 | Soluble micropin containing slightly solubility contraceptive and preparation method thereof |
| CN108785244B (en) * | 2018-06-20 | 2021-05-18 | 华中科技大学 | Hydrophobic drug-loaded soluble microneedle and preparation method thereof |
| CN109010258B (en) * | 2018-08-30 | 2021-07-13 | 上海交通大学 | Preparation method of melatonin microneedle patch |
| CN109674737B (en) * | 2019-01-07 | 2021-05-18 | 华中科技大学 | Water-soluble small molecule-based rapidly-dissolvable microneedle, and preparation and application thereof |
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