CN113750033B - Baicalin ethosome-loaded soluble hyaluronic acid microneedle array and preparation method and application thereof - Google Patents
Baicalin ethosome-loaded soluble hyaluronic acid microneedle array and preparation method and application thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7023—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms
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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
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
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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
Abstract
The invention provides a soluble hyaluronic acid microneedle array carrying baicalin ethosome, a preparation method and application thereof. The invention provides a soluble microneedle array, which takes hyaluronic acid as a matrix and carries baicalin ethosome, wherein the mass ratio of the hyaluronic acid to the baicalin ethosome is (50-100) (1-5). The soluble microneedle array prepared by the invention has the advantages of low cost, good biocompatibility, high safety, good mechanical strength, excellent transdermal drug delivery effect and good application prospect.
Description
Technical Field
The invention belongs to the field of biological medicine, and in particular relates to a soluble hyaluronic acid microneedle array carrying baicalin ethosome, a preparation method and application thereof.
Background
Psoriasis is a common chronic and easily recurrent inflammatory skin disease with characteristic skin lesions, and is mostly brownish red plaque, clear in boundary, inflammatory halation around the skin, obvious in basal infiltration and covered with multiple layers of dry off-white or silvery white scales. The current research shows that psoriasis patients have abnormality in genetic factors, biochemistry, immunity, microcirculation, endocrine and the like besides skin damage. All people can be ill, and the incidence of the young and the young is about 80 percent in the young and the middle-aged. Worldwide, caucasians have a high incidence of about 1% to 3% of the general population, and yellow-colored individuals have a incidence of about 0.1% to 0.3%.
There are many studies on drugs and preparations for treating psoriasis. Baicalin is one of the main active ingredients of the baical skullcap root of the Labiatae, and has the effects of clearing heat, detoxicating, inhibiting bacteria, resisting inflammation and the like. It has been found that baicalin can act as a therapeutic agent for psoriasis by inhibiting neutrophil (PMN) chemotaxis caused by leukotriene B4 (LTB 4), blocking LTB4 binding to PMN membrane-specific receptors. In addition, in many pharmaceutical dosage forms, external preparations are getting more attention due to the advantages of convenient use, high safety and the like, but the barrier effect of the skin barrier has a great influence on the exertion of the drug effect. Therefore, how to prepare a pharmaceutical preparation by taking baicalin as an active ingredient, the baicalin can effectively penetrate through skin barriers in the actual treatment process, target to focus positions and accumulate effective doses, and the problem still needs to be solved when the pharmaceutical preparation is applied to clinical treatment.
In order to penetrate the skin barrier, researchers have continuously studied and improved transdermal drug delivery formulations such as liposomes, etc., but conventional liposome drug delivery systems are still easily blocked by the outermost lipophilic stratum corneum of the skin, and the transdermal effect is not ideal. Based on the research of traditional liposome, the ethosome is used as a novel percutaneous administration preparation carrier, and compared with the common liposome, the ethosome has the advantages of small particle size, stable structure, better flexibility and the like, can form lipid vesicles with better permeability and encapsulation rate, and has good stability and skin tolerance.
In addition to improvements in transdermal formulations, microneedles can be used to directly pierce the stratum corneum barrier of the skin to provide a micro-tunnel for drug delivery. The microneedle array is a novel minimally invasive drug delivery mode in the field of biological medicine, and is prepared from a series of needle-like structures (namely microneedles arranged in an array form) with the length of 25-2000 microns in an array form, and can be prepared from metal, silicon, high polymer materials and the like. The medicine is directly absorbed into blood through capillary vessels through a plurality of micro-tunnels caused by the micro-needle array, so that the first pass effect is avoided, the problem of swallowing of a patient, the problem of low medicine utilization rate when the medicine passes through the gastrointestinal tract and the problem of medicine stability are avoided, the problem of patient compliance is improved, and the medicine is a research hot spot of local transdermal administration at present.
Depending on the manner of administration of the microneedle array, it can be broadly divided into: solid microneedle arrays, coated microneedle arrays, hollow microneedle arrays, soluble microneedle (gel microneedle) arrays. Wherein, the solid microneedle array is generally prepared from a metal material and a non-degradable polymer, and the administration mode comprises the steps of firstly using microneedles to puncture the skin surface to form micro-tunnels; and then the medicine is applied to the puncture part of the micro needle, and the medicine passively permeates into the skin through the pore canal.
For example, cheng Liang (SIN-HC 1 "microneedle array-delivery ethosome" combined large-flux transdermal drug delivery law study [ D ]. Northwest university, 2013) proposes a novel transdermal drug delivery system combining microneedle array-ethosome, wherein a metallic solid microneedle array is used for breaking the physical and lipid barriers of the stratum corneum, and then gentiopicroside and sinomenine hydrochloride are used as model drugs to prepare the delivery ethosome, so that the transdermal effect is verified, and the improvement of the transdermal effect is found. The solid microneedle has excellent mechanical property and is beneficial to breaking the stratum corneum barrier; however, the metal solid mechanical microneedle array is insoluble and non-degradable, and once broken or damaged in the administration process, the needle body is remained in the skin, so that potential safety hazards are caused, and complex and precise equipment is required for manufacturing, so that the preparation process is complex. The transdermal drug delivery scheme needs to treat skin by using a microneedle array and then deliver drug by using an ethosome, is complex in operation, is inconvenient and is difficult to popularize and apply widely.
The coated microneedle array is similar to the solid microneedle array, and in order to simplify the administration step, the medicine is wrapped on the surface of the microneedle body, and the medicine is dissolved and released along with the penetration of the microneedle into the skin, but the specific surface area of the microneedle is limited, so that the medicine carrying capacity of the coated microneedle array is severely limited. The hollow microneedle array is equivalent to a micron-sized injector, and the liquid medicine in the hollow hole is injected under the skin after penetrating the skin, however, the hollow structure of the hollow microneedle leads to more complex preparation process, and the inner cavity of the needle body is easy to be blocked when the hollow microneedle array is inserted into the skin due to the compactness of dermal tissue, so that the transfer of the medicine is affected.
The soluble microneedle array is mainly prepared from water-soluble polymers, the drugs are distributed on a needle point matrix, and after penetrating into the skin, the needle point absorbs a small amount of tissue fluid to dissolve and release the drugs, and the soluble microneedle array has the advantages of simple preparation method, high efficiency of drug delivery, stable drug assurance, high safety and the like. However, microneedle arrays prepared from water-soluble polymers tend to have poor mechanical properties and are difficult to effectively disrupt the skin barrier to promote transdermal drug delivery.
Therefore, the soluble drug-carrying microneedle array which has low cost, good biological safety, mass production and application, good mechanical property and excellent transdermal drug delivery effect is provided, and has very important significance.
Disclosure of Invention
The invention aims to provide a soluble drug-loaded microneedle array with excellent transdermal drug delivery effect for treating psoriasis.
The invention provides a soluble microneedle array, which takes hyaluronic acid as a matrix and carries baicalin ethosome, wherein the mass ratio of the hyaluronic acid to the baicalin ethosome is (50-100) (1-5);
the baicalin ethosome is prepared from the following raw materials: baicalin, carrier material, oil phase solvent, aqueous phase solvent, emulsifying agent and surfactant;
the carrier material is one or more of soybean lecithin, PLGA or PEG-PLGA;
the oil phase solvent is one or more of ethanol, glycerol, 1, 2-propylene glycol or polyvinyl alcohol;
the aqueous phase solvent is one or more of distilled water, PBS buffer solution and PVA aqueous solution;
the emulsifier is one or more of isopropyl palmitate, isopropyl myristate or isopropyl acrylate;
the surfactant is one or more of Tween-80, PEG-polyoxyethylene castor oil or polyethylene glycol.
Preferably, the carrier material is soy lecithin; the oil phase solvent is 1, 2-propylene glycol; the aqueous phase solvent is PBS buffer solution; the emulsifier is isopropyl myristate; the surfactant is Tween-80.
Further, the mass ratio of the hyaluronic acid to the baicalin ethosome is (80-100): (1-3), preferably 100:1.
Further, the mass volume ratio of the baicalin, the carrier material, the oil phase solvent, the emulsifier, the surfactant and the water phase solvent is as follows: 7 (40-60): 100-200): 40-80): 2-10 mg/mg/mu L/mL; preferably 7:50:200:100:60:8 mg/mg/. Mu.L/mL.
Further, the molecular weight of the hyaluronic acid is 5Kda and 40Kda, wherein the mass percentage of the hyaluronic acid with the molecular weight of 5Kda is 30% -70%; the particle size of the baicalin ethosome is 100-200 nm.
Further, the microneedles of the microneedle array are of a regular rectangular pyramid structure, the height of the rectangular pyramid is 600 μm, the width of the bottom is 290 μm, and the distance between the tips of two adjacent microneedles is 800 μm.
The invention also provides a preparation method of the soluble microneedle array, which comprises the following steps:
(1) Preparation of baicalin ethosomes: uniformly mixing the metered carrier material, the oil phase solvent, the emulsifying solvent and the surfactant to obtain an oil phase; dissolving the measured baicalin in a water phase solvent to obtain a water phase, adding the water phase into the oil phase, uniformly dispersing, and homogenizing to obtain baicalin ethosomes;
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution: uniformly mixing metered hyaluronic acid, the baicalin ethosome obtained in the step (1) and distilled water to obtain a baicalin ethosome-hyaluronic acid aqueous solution;
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array: standing the baicalin ethosome-hyaluronic acid aqueous solution obtained in the step (2), and then coating the solution on the surface of a PDMS microneedle array mould; the height of any rectangular pyramid of the PDMS microneedle array mold is 600 micrometers, and the bottom width is 290 micrometers; the tip distance of the adjacent 2 rectangular pyramids is 800 μm; and (5) carrying out vacuum treatment and normal pressure drying, and then stripping the PDMS microneedle array mould to obtain the soluble microneedle array.
Further, the rate of addition of the aqueous phase to the oil phase in step (1) was 200. Mu.L.min -1 ～500μL·min -1 The method comprises the steps of carrying out a first treatment on the surface of the The homogenizing pressure is 1-200 bar, and the homogenizing time is 5-30 min; preferably, the homogenizing pressure is 100-200 Bar, and the homogenizing time is 15-30 min;
and/or the smearing amount of the baicalin ethosome-hyaluronic acid aqueous solution in the step (3) is 100-1000 mu L; the standing time is 2-12 h, preferably 8-12 h; the vacuum treatment is carried out for 2-5 min under the vacuum environment of minus 0.01 to minus 0.1Mpa, the normal pressure drying temperature is 10-30 ℃, preferably 20-30 ℃ and the drying time is 10-15 h.
The invention also provides application of the soluble microneedle array in preparation of a medicament for treating psoriasis; preferably, the medicament for treating psoriasis is a medicament for inhibiting thymus atrophy, and/or spleen enlargement, and/or reducing inflammatory factor expression.
The invention also provides a medicine, which is a preparation prepared by adding pharmaceutically acceptable auxiliary materials or auxiliary components into the soluble microneedle array; preferably, the formulation is an external formulation; more preferably, the external preparation is a patch.
The beneficial effects of the invention include:
the hyaluronic acid used for the soluble microneedle array is a polymer material which has good biocompatibility, is soluble and water-soluble, is low in cost and is easy to obtain, and the preparation and the application of the microneedle array are facilitated to be enlarged.
The preparation process is simple, the microneedle arrays with different concentrations, regular structures and different hyaluronic acid molecular weights can be prepared under the conditions of normal temperature and negative pressure, the whole process is short in time consumption and high in repeatability, and the mass production is facilitated.
The invention provides a soluble microneedle array, which not only increases the mechanical strength of the microneedle array, but also enables the microneedle array to have the application of treating psoriasis.
The hyaluronic acid soluble microneedle array carrying baicalin ethosome provided by the invention has obvious treatment effect after treatment of mice psoriasis induced by imiquimod, and is suitable for treatment of psoriasis.
The preparation method of the microneedle array combines the preparation of baicalin ethosome, the preparation of baicalin ethosome-hyaluronic acid aqueous solution and the preparation of baicalin ethosome-hyaluronic acid loaded microneedle array, and finally the baicalin ethosome-hyaluronic acid loaded microneedle array is obtained. In the preparation of baicalin ethosome, the medicine-lipid ratio, the PH value of PBS buffer solution, the homogenization power of the ethosome, the homogenization time and the like all influence the particle size, the encapsulation efficiency and the like of the baicalin ethosome; in the aqueous solution of hyaluronic acid carrying baicalin ethosome, the molecular weight and the like of hyaluronic acid also play a key role in the preparation molding of the aqueous solution of hyaluronic acid of baicalin ethosome and the mechanical strength of the microneedle; in the preparation step of the hyaluronic acid microneedle array carrying baicalin ethosomes, the application amount of the aqueous solution containing baicalin ethosomes, the vacuum degree in a vacuum environment, the drying temperature, the drying time and the like are all used for affecting the properties of the prepared hyaluronic acid microneedle array carrying baicalin ethosomes; the loading of baicalin in the microneedle can also influence the transdermal drug delivery effect and the therapeutic effect. The invention improves and optimizes the prescription process in the aspects, finally obtains the baicalin ethosome hyaluronic acid microneedle array with regular structure, high drug loading capacity and excellent mechanical property, can efficiently and transdermally administer, and has very good application prospect as an external preparation for treating psoriasis.
Description of the terminology:
PLGA: polylactic acid-glycolic acid copolymer; PEG-PLGA: polyethylene glycol-polylactic acid-glycolic acid copolymer; PEG-40-polyoxyethylated castor oil: polyethylene glycol-40-polyoxyethylene hydrogenated castor oil refers to a block copolymer of polyethylene glycol 40 and polyoxyethylene hydrogenated castor oil.
In summary, the invention improves the preparation process of the microneedle array and simplifies the existing method for preparing the microneedle array: under the conditions of normal temperature and negative pressure, the micro-needle array with different concentrations, regular structure and different hyaluronic acid molecular weights can be prepared, the whole process is short in time consumption and strong in repeatability, and the mass production is facilitated. And (3) preparing a PDMS model with the same specification, pouring the hyaluronic acid solution on a die, and selecting a vacuum drying mode to prepare the drug-loaded microneedle array. The complex manufacturing modes of the micro-needles such as metal micro-needles and roller micro-needles are reduced, and batch production can be realized according to the number of the models. The invention combines nanometer drug delivery technology, and has targeting property compared with traditional micro-needle drug delivery. The physical and lipid barriers of the stratum corneum are firstly broken by using the soluble microneedle array with excellent mechanical strength, and simultaneously, the drug passes through the active epidermis layer in a large flux by using the hydrophilic characteristic of the ethosome, so that the transdermal effect of the drug is improved.
The mass volume ratio of baicalin, carrier material, oil phase solvent, emulsifier, surfactant and water phase solvent is as follows: 7 (40-60): (100-200): (40-80): (2-10) mg/mg/mu L/mL) means: the usage amount of baicalin corresponding to each 2-10 mL of water phase solvent is 7mg, the usage amount of corresponding carrier material is 40-60 mg, the usage amount of corresponding oil phase solvent is 100-200 mu L, the usage amount of corresponding emulsifier is 100-200 mu L, and the usage amount of corresponding surfactant is 40-80 mu L. The proportion is not limited to the combination of 7mg baicalin, 40-60 mg carrier material, 100-200 mu L oil phase solvent, 100-200 mu L emulsifier, 40-80 mu L surfactant and 2-10 mL water phase solvent, but also includes the combination of the raw material dosage which is multiplied or reduced. For example, "70mg of baicalin, 0.4 to 0.6g of carrier material, 1 to 2mL of oil phase solvent, 1 to 2mL of emulsifier, a combination of 0.4 to 0.8mL of surfactant and 20 to 100mL of aqueous phase solvent", "0.7mg of baicalin, 4 to 6mg of carrier material, 10 to 20. Mu.L of oil phase solvent, 10 to 20. Mu.L of emulsifier, 4 to 8. Mu.L of surfactant and 0.2 to 1mL of aqueous phase solvent", "0.7g of baicalin, 4 to 6g of carrier material, 10 to 20mL of oil phase solvent, 10 to 20mL of emulsifier, 4 to 8mL of surfactant and 0.2 to 1L of aqueous phase solvent", and the like.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 shows a distribution diagram of particle size of (a) baicalin ethosomes and (b) a transmission electron microscope scan of baicalin ethosomes.
FIG. 2 is an optical microscope image and a visual appearance image of the baicalin ethosome hyaluronic acid microneedle array prepared in (a); (b) A scanning electron microscope image of the prepared baicalin ethosome hyaluronic acid microneedle array and a scanning electron microscope image of a single microneedle of the prepared baicalin ethosome hyaluronic acid microneedle array; (c) Inverted fluorescence microscopy images of hyaluronic acid microneedle arrays prepared with rhodamine B instead of baicalin ethosomes.
Fig. 3 is a graph showing the effect of (a) testing the puncture strength of a hyaluronic acid microneedle array prepared by rhodamine B instead of baicalin ethosome in the isolated skin of a mouse, (B) a microscopic image of the isolated skin of the mouse under the observation of an inverted fluorescence microscope in a bright field, and (c) a microscopic image of the isolated skin of the mouse under the observation of an inverted fluorescence microscope in a fluorescence environment.
Fig. 4 shows fluorescence observation of a hyaluronic acid microneedle array prepared by rhodamine B instead of baicalin ethosome under laser confocal microscope observation on mouse isolated skin, wherein a place with the strongest red fluorescence is taken as an initial surface, fluorescent skin on an X-Y surface is collected, and fluorescence can be seen at a position 300 μm below the skin.
Fig. 5 shows the cumulative release amount of (a) the in vitro release test and the cumulative transdermal penetration amount of (b) the in vitro membrane transdermal test of the hyaluronic acid microneedle array.
Fig. 6 is a graph showing the therapeutic effect of a hyaluronic acid microneedle array loaded with baicalin ethosomes on imiquimod-induced psoriasis-like mice, and fig. 6 (a) is a graph showing the back skin changes during the experiment of mice of each group, demonstrating the success of the modeling of the experiment. The treatment of the microneedle array administration group is superior to that of baicalin raw material administration and baicalin ethosome administration. Fig. 6 (b) shows the change in body weight of mice in each group, demonstrating that imiquimod inhibits body weight of mice. FIG. 6 (c) shows organ indexes measured in spleen and thymus of mice, and the results demonstrate that the treatment with baicalin microneedle arrays can be significantly different from that of the model group.
FIG. 7 shows the expression of inflammatory factors TNF- α, IL-23, IL-2, IL-17A in mouse skin as determined by ELISA.
Detailed Description
The raw materials and equipment used in the invention are all known products and are obtained by purchasing commercial products.
EXAMPLE 1 preparation of the soluble microneedle array of the present invention
(1) Preparation of baicalin ethosome
Mixing soybean lecithin 250mg, 1, 2-propylene glycol 1.0mL and isopropyl myristate 0.5mL in EP tube, homogenizing at 45deg.C, 100deg.C for 100 r.min -1 Under the condition of weighing 10mg of baicalin raw material medicine, dissolving in oil phase, magnetically stirring to complete dissolution, adding 300 mu L of Tween-80, and continuously stirring and uniformly mixing. Into the oil phase at 500. Mu.L.min -1 8mL of aqueous phase, which is PBS buffer,the pH was 7.2 to ensure uniform dispersion in the oil phase and stirring was carried out for 1 hour after all additions. Pouring the solution prepared by stirring into a high-pressure homogenizer, wherein the power of the homogenizer is 160bar; homogenizing time is 25min. Homogenizing to obtain yellow transparent clear solution to obtain baicalin ethosome.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
uniformly dispersing hyaluronic acid into 1mL solution containing baicalin ethosomes to obtain aqueous solution of hyaluronic acid containing baicalin ethosomes, wherein the concentration of baicalin ethosomes in the aqueous solution of hyaluronic acid containing baicalin ethosomes is 1 mg.mL -1 The concentration of the hyaluronic acid is 100 mg.mL -1 The average molecular mass of the hyaluronic acid is 5Kda and 40Kda, and the mass ratio is 5Kda to 40 kda=3:7.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
standing the baicalin ethosome-hyaluronic acid aqueous solution obtained in the step (2) for 12 hours, and then, taking 1000 mu L of the baicalin ethosome-hyaluronic acid aqueous solution obtained in the step (2) to be smeared on the surface of a PDMS microneedle array die; placing the PDMS microneedle array template in a vacuum environment with the vacuum degree of 26 ℃ and 0.08 to 0.1MPa for 5 minutes to enable the hyaluronic acid solution to fully enter the gaps of the microneedle array mould, then further curing and drying the template in a vacuum drying box with the vacuum temperature of 26 ℃ under normal pressure for 12 hours, and then stripping the PDMS microneedle array mould by using tweezers to obtain the soluble hyaluronic acid microneedle array loaded with baicalin ethosome.
EXAMPLE 2 preparation of the soluble microneedle array of the present invention
(1) Preparation of baicalin ethosome
The amount of baicalin raw material is 20mg, the amount of water phase is 8mL, the water phase is PBS buffer solution, the pH is 7.2, and the rest steps are the same as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
uniformly dispersing hyaluronic acid into 1mL solution containing baicalin ethosome to obtain transparent solution containing baicalin ethosomeAqueous solution of hyaluronic acid, wherein the concentration of baicalin ethosome in the aqueous solution of hyaluronic acid containing baicalin ethosome is 2 mg.mL -1 The concentration of the hyaluronic acid is 100 mg.mL -1 The average molecular mass of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=3:7.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
EXAMPLE 3 preparation of soluble microneedle arrays of the invention
(1) Preparation of baicalin ethosome
The amount of baicalin raw material is 35mg, the amount of water phase is 8mL, the water phase is PBS buffer solution, the pH is 7.2, and the rest steps are the same as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
uniformly dispersing hyaluronic acid into a solution containing baicalin ethosomes to obtain an aqueous solution of hyaluronic acid containing baicalin ethosomes, wherein the concentration of baicalin ethosomes in the aqueous solution of hyaluronic acid containing baicalin ethosomes is 3.5 mg.mL -1 The concentration of the hyaluronic acid is 100 mg.mL -1 The average molecular mass of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=3:7.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
EXAMPLE 4 preparation of soluble microneedle arrays of the invention
(1) Preparation of baicalin ethosome
The amount of baicalin raw material is 35mg, the amount of water phase is 8mL, the water phase is PBS buffer solution, the pH is 7.2, and the rest steps are the same as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
uniformly dispersing hyaluronic acid into a solution containing baicalin ethosomes to obtain an aqueous solution of hyaluronic acid containing baicalin ethosomes, wherein the aqueous solution of hyaluronic acid containing baicalin ethosomes contains yellowThe concentration of the qin glycoside ethosome is 3.5 mg.mL -1 The concentration of the hyaluronic acid is 80 mg.mL -1 The average molecular weight of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=3:7.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
EXAMPLE 5 preparation of soluble microneedle arrays of the invention
(1) Preparation of baicalin ethosome
The amount of baicalin raw material is 35mg, the amount of water phase is 8mL, the water phase is PBS buffer solution, the pH is 7.2, and the rest steps are the same as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
uniformly dispersing hyaluronic acid into a solution containing baicalin ethosomes to obtain an aqueous solution of hyaluronic acid containing baicalin ethosomes, wherein the concentration of baicalin ethosomes in the aqueous solution of hyaluronic acid containing baicalin ethosomes is 3.5 mg.mL -1 The concentration of the hyaluronic acid is 90 mg.mL -1 The average molecular weight of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=3:7.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
EXAMPLE 6 preparation of the soluble microneedle array of the invention
(1) Preparation of baicalin ethosome
The procedure is as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
the average molecular mass of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=2:8. The rest of the procedure is the same as in example 3.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
EXAMPLE 7 preparation of soluble microneedle arrays of the invention
(1) Preparation of baicalin ethosome
The procedure is as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
the average molecular mass of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=1:9. The rest of the procedure is the same as in example 3.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
EXAMPLE 8 preparation of the soluble microneedle array of the invention
(1) Preparation of baicalin ethosome
The procedure is as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
the average molecular mass of the hyaluronic acid is 5Kda and 40Kda, and the ratio of the hyaluronic acid to the 40 kda=4:6. The rest of the procedure is the same as in example 3.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
Comparative example 1 preparation of soluble microneedle array without baicalin ethosome
(1) Preparation of baicalin ethosome
Baicalin content is 0mg, and the rest steps are the same as in example 1.
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution:
the concentration of baicalin ethosome is 0mg/mL, and the rest steps are the same as in example 1.
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array:
the procedure is as in example 1.
Test example 1 prescription Process optimization of baicalin ethosome
Referring to the preparation method of the step (1) of the example 1, the raw material dosage ratio, the water phase pH, the addition amount of Tween-80 and the homogenizing power are adjusted by a single factor, a plurality of baicalin ethosomes are prepared, and the particle size, the encapsulation efficiency and the drug loading rate of the baicalin ethosomes are detected and calculated, so that the results are shown in the table 1.
TABLE 1
Note that: the exterior traditional Chinese medicine lipid ratio specifically refers to the mass ratio (mg/mg) of baicalin as the medicine and lecithin as the carrier material.
According to the results, the medicine has a medicine-fat ratio of 7:50, an aqueous phase pH of 7.2: tween-80 mass-volume ratio 7:60, medicine: (oil phase solvent+emulsifier) mass-to-volume ratio of 7:300, homogenizing pressure power 160Bar as a preferred parameter (i.e., protocol of example 3), three batches of baicalin ethosomes were prepared and characterized, and the results are shown in table 2:
TABLE 2
Therefore, the baicalin ethosome prepared under the optimized preparation parameters has high encapsulation efficiency and drug loading capacity and good stability.
Experimental example 2, preparation of microneedle array prescription Process optimization
Referring to the preparation method of the step (2) of the example 1, transparent microneedle arrays were prepared by uniformly dispersing hyaluronic acid in a solvent containing no baicalin ethosome, and the influence of the type and amount of the solvent and the molecular weight of the hyaluronic acid on the performance of the microneedle arrays was examined:
1. solvent screening
The types of solvents added to disperse hyaluronic acid into baicalin ethosome solution were selected, and the results were as follows:
it can be seen that the distilled water is used as the solvent of baicalin ethosome-hyaluronic acid, and the molding property of the microneedle array is best further, and no bubbles are generated basically, so that the influence of the water consumption on the performance of the microneedle array is further verified by using the water as the solvent. In the preparation of the baicalin-carrying ethosome hyaluronic acid microneedle array, an aqueous solution (e.g., PBS buffer solution, PVA aqueous solution, etc.) without ethanol is used as a dispersion.
2. Water addition amount
As can be seen, when the water addition amount is 1ml, the prepared microneedle array has good formability, high needle content and excellent mechanical strength. Thus, in the process of preparing baicalin ethosome-loaded hyaluronic acid microneedle arrays, a preparation step of dispersing hyaluronic acid into 1ml of baicalin ethosome solution was adopted.
3. Hyaluronic acid molecular weight
It can be seen that when 5kDa/40kDa molecular weight is used as hyaluronic acid, the moldability and hardness are more excellent than those of hyaluronic acid of other molecular weights.
4. Mechanical Property investigation of the molecular weight ratio of hyaluronic acid
It can be seen that hyaluronic acid with a molecular weight of 5kDa/40kDa is found at 5kDa: the 40kDa ratio is 3:7-7: 3 and has good formability and excellent mechanical properties.
Experimental example 3 optimization of microneedle array preparation Process
After the solution for preparing the microneedle array is obtained, the solution is smeared on the surface of a microneedle array die, and the process for forming the microneedle array comprises the following steps of: (1) The time for vacuum pumping is performed in order to make the solution fully enter the gaps of the microneedle array mold; and (2) drying and curing times, the results were as follows:
1. influence of vacuuming time on formability and needle content:
it can be seen that the solution can be fully introduced into the gaps of the microneedle array mold by placing under vacuum for 5 minutes, and the microneedle array mold has good formability, preferably excellent formability at the time of 5 minutes, and the highest number of microneedles. However, the vacuum time is too long, and up to 10 minutes may cause the liquid medicine to overflow from the mold or cause evaporation of water, resulting in failure to form the microneedles. Thus, a vacuum treatment time of 5min was taken during the preparation of the microneedle array.
2. Effect of dry cure temperature on microneedle array:
the above results indicate that the dry microneedle array has good moldability at a low temperature (30 ℃ or lower) and fewer bubbles, and is a preferable dry curing condition. Wherein, the drying is carried out for 12 hours plus or minus 1 hour at the constant temperature of 26 ℃, the molding is good, the bubbles are few, and the most preferable experimental scheme is adopted.
Experimental example 4 characterization of baicalin ethosomes and soluble microneedle arrays of the invention
The particle size and PDI of baicalin ethosomes of example 3 were measured by a malvern particle sizer to obtain fig. 1 (a); taking 0.3mL of the prepared baicalin ethosome, adopting an ultrafiltration centrifuge tube with a molecular weight of 3K, and sucking the free medicine at the lower layer after high-speed refrigerated centrifugation (centrifugal force 16000r,4 ℃ for 15 min). Filtering, sampling, and calculating the peak area. Substituting the drug-loading rate into a standard curve to obtain the encapsulation efficiency and the drug-loading rate, wherein the calculation method comprises the following steps:
encapsulation efficiency EE (%) = (encapsulated drug/total drug dose) ×100%;
drug loading DL (%) = (total weight of encapsulated drug/ethosome) ×100%;
chromatographic conditions: the column was Swell C-18 (150 mm. Times.4.6 mm,5 μm); mobile phase a:40% aqueous methanol; mobile phase B:60% phosphoric acid aqueous solution; volume flow 1ml/min; a detection wavelength of 278nm; the sample injection amount is 10 mu L; column temperature was 30 ℃. The theoretical plate number is more than or equal to 5000, and the separation degree is more than 1.5.
The transmission electron microscope scanning method comprises the following steps: taking an appropriate amount of ethosome, diluting with PBS (phosphate buffer solution) with pH7.0 for 10 times, dripping 20 mu L of the ethosome on a copper mesh, absorbing for 5min, adding an appropriate amount of 1% phosphotungstic acid for dyeing for 1min, naturally air-drying, observing the shape under a Transmission Electron Microscope (TEM), and photographing. FIG. 1 (b) is obtained.
The appearance of the soluble microneedle array was observed, and the appearance was observed with an optical microscope and a scanning electron microscope. Because baicalin has weak fluorescence, rhodamine B with fluorescence is selected to replace baicalin ethosome for preparing a transparent microneedle array in the experiment, and the transparent microneedle array is observed by a fluorescence microscope. The shape, size, height, etc. of the microneedles can be observed in a fluorescent environment.
2. Experimental results
As can be seen from the graph (a) of FIG. 1, the particle size of the baicalin ethosome prepared by experiment is about 100nm, the encapsulation efficiency is 79.09% by calculation, and the drug loading is 2.77%; as can be seen from fig. 1 (b), the prepared baicalin ethosomes are spheroids and uniformly dispersed; the portable microscopic image of the baicalin ethosome microneedle array prepared in fig. 2 (a) and the scanning electron microscope image of fig. 2 (B) can observe that the needle type of the microneedle array is perfect, rhodamine B with fluorescence is used for replacing baicalin ethosome to prepare transparent microneedles, and the needle points of all the microneedles of the microneedle array are perfect and uniformly distributed as observed under an inverted fluorescence microscope, wherein the microneedles are in a regular pyramid shape, the height of a rectangular pyramid is 600 mu m, and the bottom width is 290 mu m; the tip distance of the adjacent 2 rectangular pyramids was 800 μm. The following conclusions are explained: the baicalin ethosome prepared by the experiment has high entrapment rate and drug loading capacity and small particle size, and the microneedle tips of the prepared microneedle array are intact and distributed uniformly. The method for preparing the baicalin ethosome hyaluronic acid microneedle array by the experiment is proved to be feasible, and the medicine carrying capacity and the encapsulation efficiency of the baicalin ethosome are high.
Experimental example 5 verification of skin penetration effect of hyaluronic acid microneedle array of the invention
1. Experimental method
The mice are killed, dehaired and frozen for later use, the mouse skin is thawed before the experiment, soaked in physiological saline, separated into squares of 1.5cm X1.5 cm by a medical scalpel, and the fat layer is removed. The prepared patch of microneedle array (example 3) was inserted into the isolated skin for 5min, then removed, and the skin surface was rubbed with filter paper, and the state of the microneedle array on the skin was observed with a portable microscope and an inverted fluorescence microscope. Rhodamine B was used in place of baicalin ethosomes in the microneedle array of example 3, and observation was performed using an inverted fluorescence microscope.
2. Experimental results (subsequent experimental results were all performed with the microneedle prepared in example 3)
From fig. 3, it can be seen that after the microneedle array pierces the skin, obvious pores are left, the pores are uniformly distributed among the pores (fig. 3 (a)), and the distribution of the microneedle array in the skin is observed by an inverted fluorescence microscope under a bright field state and a fluorescence state, the microneedle array appears pink (fig. 3 (b)), and the microneedle array emits light blue luster under the fluorescence condition (fig. 3 (c)). Fig. 4 shows fluorescence observation of a hyaluronic acid microneedle array prepared by rhodamine B instead of baicalin ethosome under laser confocal microscope observation on mouse isolated skin, wherein a place with the strongest red fluorescence is taken as an initial surface, fluorescent skin on an X-Y surface is collected, and fluorescence can be seen at a position 300 μm below the skin. The following conclusions are explained: the micro needle prepared by the experiment can penetrate through the skin stratum corneum, and has good penetrating performance.
Experimental example 6 release and transdermal Effect of baicalin ethosome in microneedle arrays of the present invention
1. Experimental method
In vitro release experiments: simulating human body environment, taking physiological saline as a release medium, carrying out in vitro research on the prepared microneedle array, loading the baicalin ethosome-hyaluronic acid aqueous solution prepared in the example 3 into a dialysis bag, carrying out water bath at 37 ℃ in 50mL of physiological saline, carrying out dynamic dialysis at 60r/min, and supplementing 1mL of physiological saline at the same temperature while sucking 1mL of physiological saline at 0.5, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18 and 24h, and measuring the content of free baicalin in the dialysis medium, thereby calculating the cumulative release rate of baicalin liposome nanoparticles.
Cn＝(M1+M2+···+Mn)/M Total (S) *100％
Where Cn is the cumulative release rate, M is the release amount of theoretical 100% release, mn is the release amount obtained at each time point, mn=c×v
In vitro transdermal experiments: experiments were performed using a Franz transdermal diffusion tester. The ethosome-microneedle array prepared in example 3 was tightly attached to a cellulose acetate film, and then a backing gel layer prepared before was attached, and the receiving tank was filled with physiological saline as a receiving medium, and the temperature was set at 32℃and the rotational speed was set at 200r/min. And respectively sucking 500 mu L of the baicalin ethosome-microneedle array in 0,2,4,6,8, 10, 12h,24h and 48h, simultaneously supplementing an equal amount of isothermal fresh normal saline, filtering with a 0.22 mu m microporous filter membrane, sampling, calculating cumulative permeation quantity (Qn) according to the following formula, and measuring the cumulative permeation quantity of the baicalin ethosome-microneedle array in 48 h.
Qn＝(Cn×V)/A Cn＝C1+C2+……+Cj
In the above formula, qn is the cumulative permeation per unit area (g/cm 2 ) Cn is the drug concentration (g/mL) measured at the nth time point, V is the sampling volume (mL), A is the receiving well area (cm) 2 ). In this experiment, v=15ml, a=1.54 cm 2 。
2. Experimental results
As shown in fig. 5, the cumulative release amount of the hyaluronic acid microneedle array was 57.43%, the cumulative membrane permeation amount was 77.93%, and the release rate was slower after 12 hours as compared with the release of the drug substance. The cumulative permeation quantity of the 3 preparations in unit area of membrane permeation within 48 hours is calculated, so that the release of the two dosage forms is gradually gentle before 48 hours, and the drugs of the microneedle group slowly permeate the acetate membrane, thereby playing a slow-release effect.
Experimental example 7 therapeutic Effect of the soluble microneedle array of the present invention on psoriasis
1. Experimental method
Pathological skin of mice with imiquimod-induced psoriasis:
the method is divided into six groups by a random grouping method, and each group comprises six. Blank control group, model group, ethosome group, microneedle group, positive drug control group and raw drug control group, and is suitable for one week. BALB/C mice were dehaired on their backs with a smooth and atraumatic dehaired area of about 1.5 cm. Times.1.5 cm. BALB/C mice were coated with imiquimod at 62.5 mg/(cm) 2 Day), fig. 6a demonstrates molding success.
The microneedle array of example 3 was administered daily beginning on day three afternoon, pressed against the pathological skin of psoriatic mice for 5min, and a backing adhesive gel layer was further administered to fix the microneedle array to the back of the mice, each treatment lasting for 4h, for 5 consecutive days, after day 8, the mice were sacrificed after weighing, the back skin, spleen tissue and thymus tissue of the mice were removed, and HE staining was performed on the skin of the mice. Expression of inflammatory factors TNF- α, IL-23, IL-2, IL-17A in mouse skin as determined by ELISA.
2. Experimental results
As shown in fig. 6b, imiquimod was demonstrated to have an inhibitory effect on the body weight of mice, and resulted in thymus atrophy (thymus as an immune organ, atrophy of which reflects a decrease in immune function), splenomegaly (indicating inflammation generation).
As shown in fig. 6c, the organ index changes measured in spleen and thymus of the mice prove that the baicalin ethosome microneedle array can effectively inhibit splenomegaly and thymus atrophy caused by imiquimod induction, and the treatment group of the baicalin microneedle array has obvious differences compared with the model group, and the effect of the baicalin ethosome is further improved compared with that of the baicalin bulk drug and the baicalin ethosome.
From fig. 7, it can be seen that the soluble microneedle array of the present invention has a remarkable effect of inhibiting typical psoriasis inflammatory factors and exhibits an enhancement effect compared to baicalin ethosomes alone.
The results show that the baicalin ethosome microneedle array can effectively inhibit splenomegaly and thymus atrophy caused by imiquimod induction, reduce the expression of inflammatory factors characteristic of psoriasis, and has great potential for being applied to psoriasis treatment.
In conclusion, the invention provides the soluble microneedle array for treating psoriasis, which has low cost and simple preparation method and is suitable for industrialized mass production; the microneedle array has the advantages of good biocompatibility, high safety, good mechanical strength, excellent transdermal drug delivery effect, and good application prospect, and can be used for treating psoriasis.
Claims (10)
1. The soluble microneedle array is characterized in that hyaluronic acid is used as a matrix and loaded with baicalin ethosomes, and the mass ratio of the hyaluronic acid to the baicalin ethosomes is 100:1, a step of;
the baicalin ethosome is prepared from the following raw materials: baicalin, carrier material, oil phase solvent, aqueous phase solvent, emulsifying agent and surfactant;
the carrier material is soybean lecithin; the oil phase solvent is 1, 2-propylene glycol; the aqueous phase solvent is PBS buffer solution; the emulsifier is isopropyl myristate; the surfactant is Tween-80; the mass volume ratio of baicalin, carrier material, oil phase solvent, emulsifier, surfactant and water phase solvent in the baicalin ethosome is as follows: 7mg (40-60) mg (100-200) mu L (40-80) mu L (2-10) mL;
the molecular weight of the hyaluronic acid is 5kDa and 40kDa, wherein the hyaluronic acid with the molecular weight of 5kDa accounts for 30% of the total amount of the hyaluronic acid by mass.
2. The soluble microneedle array of claim 1, wherein the particle size of the baicalin ethosome is 100-200 nm.
3. The soluble microneedle array of claim 1, wherein the microneedles are in a regular rectangular pyramid structure, the rectangular pyramid height is 600 μm, the bottom width is 290 μm, and the distance between adjacent microneedle tips is 800 μm.
4. A method for preparing a soluble microneedle array according to any one of claims 1 to 3, comprising the steps of:
(1) Preparation of baicalin ethosomes: uniformly mixing the metered carrier material, the oil phase solvent, the emulsifier and the surfactant to obtain an oil phase; dissolving the measured baicalin in a water phase solvent to obtain a water phase, adding the water phase into the oil phase, uniformly dispersing, and homogenizing to obtain baicalin ethosomes;
(2) Preparation of baicalin ethosome-hyaluronic acid aqueous solution: uniformly mixing metered hyaluronic acid, the baicalin ethosome obtained in the step (1) and distilled water to obtain a baicalin ethosome-hyaluronic acid aqueous solution;
(3) Preparation of baicalin ethosome-hyaluronic acid microneedle array: standing the baicalin ethosome-hyaluronic acid aqueous solution obtained in the step (2), and then coating the solution on the surface of a PDMS microneedle array mould; the height of any rectangular pyramid of the PDMS microneedle array mold is 600 micrometers, and the bottom width is 290 micrometers; the tip distance of the adjacent 2 rectangular pyramids is 800 μm; and (5) carrying out vacuum treatment and normal pressure drying, and then stripping the PDMS microneedle array mould to obtain the soluble microneedle array.
5. The process of claim 4, wherein the rate of addition of the aqueous phase to the oil phase in step (1) is 200. Mu.L min -1 ~500μL·min -1 The method comprises the steps of carrying out a first treatment on the surface of the The homogenizing pressure is 1-200 bar, and the homogenizing time is 5-30 min;
and/or the smearing amount of the baicalin ethosome-hyaluronic acid aqueous solution in the step (3) is 100-1000 mu L; the standing time is 2-12 hours; the vacuum treatment is carried out by placing for 2-5 min under the vacuum environment of-0.01 to-0.1 Mpa, the normal pressure drying temperature is 10-30 ℃, and the drying time is 10-15 h.
6. Use of a soluble microneedle array according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment of psoriasis.
7. The use according to claim 6, wherein the medicament for the treatment of psoriasis is a medicament for the inhibition of thymus atrophy, and/or splenomegaly, and/or for the reduction of inflammatory factor expression.
8. A medicament characterized in that it is a preparation prepared from the soluble microneedle array according to any one of claims 1 to 3, together with pharmaceutically acceptable auxiliary materials or auxiliary components.
9. The medicament of claim 8, wherein the formulation is an external formulation.
10. The medicament according to claim 9, wherein the external preparation is a patch.
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| CN108785244A (en) * | 2018-06-20 | 2018-11-13 | 华中科技大学 | A kind of soluble micropin and preparation method thereof supporting dewatering medicament |
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| "微针阵列—醇质体"组合下秦艽中龙胆苦苷的经皮释药规律研究;危红华;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》(第9期);E057-28 * |
| 正交试验优选黄芩苷二元醇质体制备工艺;金思岑;;中国药房(19);1777-1779 * |
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