CN116548205B - Microneedle preparation, microneedle patch, and preparation methods and applications thereof - Google Patents
Microneedle preparation, microneedle patch, and preparation methods and applications thereof Download PDFInfo
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- CN116548205B CN116548205B CN202310453651.5A CN202310453651A CN116548205B CN 116548205 B CN116548205 B CN 116548205B CN 202310453651 A CN202310453651 A CN 202310453651A CN 116548205 B CN116548205 B CN 116548205B
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- microneedle
- hyaluronic acid
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- weight hyaluronic
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- 238000002360 preparation method Methods 0.000 title claims abstract description 58
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims abstract description 79
- 229920002674 hyaluronan Polymers 0.000 claims abstract description 79
- 229960003160 hyaluronic acid Drugs 0.000 claims abstract description 79
- 102000018997 Growth Hormone Human genes 0.000 claims abstract description 48
- 108010051696 Growth Hormone Proteins 0.000 claims abstract description 48
- 239000000122 growth hormone Substances 0.000 claims abstract description 48
- 239000000463 material Substances 0.000 claims abstract description 43
- 230000008635 plant growth Effects 0.000 claims abstract description 43
- 239000002904 solvent Substances 0.000 claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000012377 drug delivery Methods 0.000 claims abstract description 15
- 239000000758 substrate Substances 0.000 claims description 32
- 239000000123 paper Substances 0.000 claims description 30
- 229930191978 Gibberellin Natural products 0.000 claims description 19
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims description 19
- 239000003448 gibberellin Substances 0.000 claims description 19
- CERQOIWHTDAKMF-UHFFFAOYSA-M Methacrylate Chemical compound CC(=C)C([O-])=O CERQOIWHTDAKMF-UHFFFAOYSA-M 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 9
- 229930192334 Auxin Natural products 0.000 claims description 8
- 239000002363 auxin Substances 0.000 claims description 8
- 239000004062 cytokinin Substances 0.000 claims description 8
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 claims description 8
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- 239000000416 hydrocolloid Substances 0.000 claims description 4
- 239000004698 Polyethylene Substances 0.000 claims description 3
- 239000011086 glassine Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000002985 plastic film Substances 0.000 claims description 3
- 229920006255 plastic film Polymers 0.000 claims description 3
- -1 polyethylene Polymers 0.000 claims description 3
- 229920000573 polyethylene Polymers 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 18
- 239000000126 substance Substances 0.000 abstract description 4
- 239000002699 waste material Substances 0.000 abstract description 4
- 239000003337 fertilizer Substances 0.000 abstract description 2
- 239000000575 pesticide Substances 0.000 abstract description 2
- 125000005395 methacrylic acid group Chemical group 0.000 abstract 2
- 241000196324 Embryophyta Species 0.000 description 41
- 210000001519 tissue Anatomy 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 10
- 239000000872 buffer Substances 0.000 description 9
- 230000002792 vascular Effects 0.000 description 9
- 239000000853 adhesive Substances 0.000 description 6
- 230000001070 adhesive effect Effects 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 206010052428 Wound Diseases 0.000 description 5
- 208000027418 Wounds and injury Diseases 0.000 description 5
- JLIDBLDQVAYHNE-YKALOCIXSA-N (+)-Abscisic acid Chemical compound OC(=O)/C=C(/C)\C=C\[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-YKALOCIXSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000000149 penetrating effect Effects 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000005507 spraying Methods 0.000 description 4
- 241000219195 Arabidopsis thaliana Species 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 235000017807 phytochemicals Nutrition 0.000 description 3
- 229930000223 plant secondary metabolite Natural products 0.000 description 3
- 210000003491 skin Anatomy 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000007921 spray Substances 0.000 description 3
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 2
- 239000005977 Ethylene Substances 0.000 description 2
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 2
- 208000002193 Pain Diseases 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- FCRACOPGPMPSHN-UHFFFAOYSA-N desoxyabscisic acid Natural products OC(=O)C=C(C)C=CC1C(C)=CC(=O)CC1(C)C FCRACOPGPMPSHN-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108010022355 Fibroins Proteins 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000004205 dimethyl polysiloxane Substances 0.000 description 1
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000003822 epoxy resin Substances 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- 208000004296 neuralgia Diseases 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 229920000647 polyepoxide Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 230000037380 skin damage Effects 0.000 description 1
- 210000000434 stratum corneum Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000013271 transdermal drug delivery Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
- A01N25/10—Macromolecular compounds
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/34—Shaped forms, e.g. sheets, not provided for in any other sub-group of this main group
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/06—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
- A01N43/12—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings condensed with a carbocyclic ring
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N45/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds having three or more carbocyclic rings condensed among themselves, at least one ring not being a six-membered ring
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Biodiversity & Conservation Biology (AREA)
- Botany (AREA)
- Toxicology (AREA)
- Forests & Forestry (AREA)
- Ecology (AREA)
- Cultivation Of Plants (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Chemical & Material Sciences (AREA)
Abstract
The invention relates to the technical field of microneedles, in particular to a microneedle preparation, a microneedle patch, a preparation method and application thereof. The microneedle preparation contains a microneedle skeleton material, a plant growth hormone and a solvent; the microneedle skeleton material contains large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylic acid-esterified hyaluronic acid, and the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylic acid-esterified hyaluronic acid is 1:2-5:0.1-0.5. The microneedle patch prepared by the microneedle preparation can be used for plant drug delivery, has a high-efficiency delivery effect, is low in preparation cost, simple and safe in use method, ensures high yield, and can reduce the input of substances such as water, chemical fertilizer and pesticide and resource waste.
Description
Technical Field
The invention relates to the technical field of microneedles, in particular to a microneedle preparation, a microneedle patch, a preparation method and application thereof.
Background
The plant growth hormone can promote the development of plant root system, promote the growth of plant stalk and promote the flowering and fruiting of plant. The artificial replenishment of exogenous plant growth hormone during plant growth can raise crop yield and quality. The current method for supplementing growth hormone to plants is two ways of foliage spraying and soil fertilization, and mainly relies on active transportation and passive transportation of plants to be absorbed from the outside. The two methods have larger resource loss, the loss rate of active substances in the foliar spraying method is 30-40%, and the active substances in the soil fertilizing method are easy to degrade in the soil. Modern agriculture is devoted to developing a high-efficiency nutrient conveying mode, and simultaneously, the high yield is ensured, and meanwhile, the investment of substances such as water, chemical fertilizers, pesticides and the like and the resource waste are reduced to the greatest extent. Therefore, there is a need to develop a completely new delivery system for precision agriculture that enables efficient low-loss delivery of phytochemicals such as micronutrients in crops.
Microneedles are generally defined as needles 10-2000 μm in height and 10-50 μm in width that can pass through just the stratum corneum of the skin without touching the pain nerve, form drug delivery channels on the skin surface to allow the drug to reach the skin to a specified depth and enter the subcutaneous capillary network to be absorbed, and act to facilitate drug penetration without causing pain and skin damage. As a novel transdermal drug delivery technology, the microneedle has the advantages of no pain, minimally invasive, convenience, safety and wide application scene. The microneedle patches in the prior art are mostly applied to animal or human body drug administration, and research on accurate delivery of plants is not reported.
Disclosure of Invention
The invention aims to solve the problems of high loss rate, environmental pollution, unreported application of a microneedle technology to plant drug delivery and the like of the conventional drug delivery method in the prior art, and provides a microneedle preparation, a microneedle patch, a preparation method and application thereof. The microneedle patch prepared by the microneedle preparation can be used for plant drug delivery, has a high delivery effect, and is low in preparation cost, simple and safe in use method.
In order to achieve the above object, a first aspect of the present invention provides a microneedle preparation comprising a microneedle skeleton material, a plant growth hormone, and a solvent; the microneedle skeleton material contains large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylate hyaluronic acid; the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:2-5:0.1-0.5.
Preferably, the mass ratio of the plant growth hormone, the microneedle skeleton material and the solvent is 1:10-300:100-3000.
Preferably, the plant growth hormone is selected from at least one of auxin, gibberellin and cytokinin.
Preferably, the solvent is water or a buffer.
In a second aspect, the present invention provides a microneedle patch comprising a microneedle array made of the microneedle formulation and a substrate, the microneedle array being disposed on the substrate.
Preferably, the microneedle patch further comprises release paper, the substrate is attached to the release paper, an opening is formed in the release paper, and the microneedle array is suitable for penetrating through the opening.
Preferably, the substrate is a hydrocolloid.
Preferably, the release paper is glassine paper or polyethylene plastic film.
Preferably, the microneedle array comprises a plurality of microneedles, and a plurality of the microneedles are arranged in a plurality of concentric circle structures.
The third aspect of the invention provides a method for preparing a microneedle patch, comprising the following steps:
s1, mixing a microneedle skeleton material, plant growth hormone and a solvent to obtain a microneedle preparation;
s2, injecting the microneedle preparation into a mould, drying, and demoulding to obtain the microneedle array;
s3, assembling the microneedle array with a substrate and optional release paper to obtain the microneedle patch;
the microneedle skeleton material contains large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylate hyaluronic acid; the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:2-5:0.1-0.5.
Preferably, in step S1, the plant growth hormone is selected from at least one of auxin, gibberellin and cytokinin; further preferably, the mass ratio of the plant growth hormone, the microneedle scaffold material and the solvent is 1:10-300:100-3000.
Preferably, the solvent is water or a buffer.
Preferably, the microneedle array comprises a plurality of microneedles, and a plurality of the microneedles are arranged in a plurality of concentric circle structures.
Preferably, in step S3, the method of assembling the microneedle array with a substrate and optionally a release paper includes: attaching the microneedle array on the substrate, attaching the release paper to the substrate, and penetrating the microneedle array out of the opening of the release paper.
The fourth aspect of the invention provides an application of the microneedle patch or the microneedle patch prepared by the preparation method as a plant drug delivery carrier.
Through the technical scheme, the invention has the beneficial effects that:
according to the microneedle preparation provided by the invention, the microneedle skeleton material is limited to the large molecular weight hyaluronic acid, the small molecular weight hyaluronic acid and the methacrylate hyaluronic acid, and the three components are limited in the mass ratio range, so that the prepared microneedle array can enable growth hormone to directly act on plant tissues and vascular systems through interaction among the large molecular weight hyaluronic acid, the small molecular weight hyaluronic acid and the methacrylate hyaluronic acid with a specific mass ratio, thereby realizing permeation absorption of the plant tissues and vascular systems on the growth hormone, achieving better delivery effect, avoiding drug loss in traditional application, and realizing application of the microneedle technology on plant drug delivery. The microneedle prepared by the microneedle preparation provided by the invention can be dissolved in plant tissues, has the advantages of buffer administration and full administration, and ensures high-efficiency and low-loss delivery of phytochemicals. The microneedle patch prepared by the microneedle preparation provided by the invention is minimally invasive, efficient and convenient to use, the production process is simple, the industrial large-scale mass production can be rapidly realized, and the microneedle patch can be widely popularized in the market.
Drawings
FIG. 1 is a schematic illustration of the structure of a single piece of the microneedle patch of the present invention;
FIG. 2 is a schematic cross-sectional view of a microneedle array according to the present invention;
FIG. 3 is a schematic diagram of a microneedle mould preparation flow, wherein A is a microneedle male mould, B is a polymer material injection male mould and curing, and C is a prepared polymer female film, namely a microneedle mould;
FIG. 4 is an SEM image of a microneedle array prepared according to example 1;
FIG. 5 is an enlarged view of FIG. 4, A being an enlarged view of a single microneedle;
fig. 6 is a schematic diagram of two specifications of the microneedle patch according to the present invention, a being a monolithic package and B being six-piece packages.
Reference numerals
1 release paper, 2 substrate, 3 microneedle array, 4 microneedles.
Detailed Description
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and are to be considered as specifically disclosed herein.
The first aspect of the invention provides a microneedle preparation, which comprises a microneedle skeleton material, plant growth hormone and a solvent, wherein the microneedle skeleton material comprises large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylic acid, and the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylic acid is 1:2-5:0.1-0.5. The inventor finds that the microneedle skeleton material is limited to contain the three components according to the mass ratio, and is mixed according to the mass ratio, and the prepared microneedle array can enable growth hormone to directly act on plant tissues and vascular systems through interaction of the three components, so that a better delivery effect is achieved, meanwhile, the traditional application drug loss is avoided, and the application of a microneedle technology in plant drug delivery is realized. Further preferably, the mass ratio of the large molecular weight hyaluronic acid, the small molecular weight hyaluronic acid and the methacrylated hyaluronic acid is 1:2-4:0.2-0.5. And hyaluronic acid in the microneedle skeleton material can be dissolved in plant tissues, so that the prepared microneedle has the advantages of buffer administration and full administration, and meanwhile, the hyaluronic acid can accelerate the repair of wounds and play a role in curing the wounds.
According to the invention, the large molecular weight hyaluronic acid is hyaluronic acid with molecular weight more than or equal to 50000da, and the small molecular weight hyaluronic acid is hyaluronic acid with molecular weight less than 50000 da.
According to the present invention, preferably, the mass ratio of the plant growth hormone, the microneedle skeleton material, and the solvent is 1:10-300:100-3000. Under the condition of the mass ratio, the administration effect of the microneedle array can be improved. From the viewpoint of further improving the administration effect of the micro-needle prepared from the microneedle preparation to plants, it is further preferable that the mass ratio of the plant growth hormone, the microneedle scaffold material, and the solvent is 1:100-200:1000-2000.
According to the present invention, the plant growth hormone may be auxin, gibberellin, cytokinin, abscisic acid, ethylene, etc., and preferably, the plant growth hormone is at least one selected from auxin, gibberellin and cytokinin. It has been found that the combination of the above plant growth hormone with the microneedle scaffold material can better allow the plant growth hormone in the microneedle preparation to act on plant growth. More preferably, the plant growth hormone is gibberellin.
According to the invention, the solvent is water or buffer solution, and the solvent is water or buffer solution, so that the microneedle skeleton material and the plant growth hormone have good dispersion effect, the plant growth hormone is uniformly released in an acting object, and the administration effect of the prepared microneedle array is improved. Further preferably, the solution is water.
A second aspect of the present invention provides a microneedle patch comprising a microneedle array 3 made of the microneedle formulation of any one of claims 1 to 3 and a substrate 2, the microneedle array 3 being disposed on the substrate 2.
According to the invention, the substrate 2 is coated with a medical adhesive, and the microneedle array 3 is connected with the substrate 2 through the medical adhesive. The microneedle array 3 contains plant growth hormone, and the microneedle array 3 can enable the growth hormone to directly act on plant tissues and vascular systems, so that the penetration and absorption of the plant tissues and vascular systems on the growth hormone are realized, the application of the microneedle technology on plant drug delivery is realized, and a better delivery effect can be achieved.
Preferably, as shown in fig. 1 and 2, the microneedle patch further comprises a release paper 1, the substrate 2 is attached to the release paper 1 by a medical adhesive, an opening is formed in the release paper 1, and the microneedle array 3 is adapted to pass through the opening.
Preferably, the substrate is a hydrocolloid which has good air permeability and is convenient for coating an adhesive, and can be attached to the rootstock of crops. After the microneedle skeleton material is degraded, the hydrocolloid substrate can be easily removed, so that the wound is convenient to heal.
Preferably, the release paper is glassine paper or polyethylene plastic film, and can protect the substrate and prevent adhesion.
Preferably, the microneedle array 3 includes a plurality of microneedles 4, and a plurality of the microneedles 4 are arranged in a plurality of concentric circular structures, which can better utilize growth hormone, avoid waste, and the diameter, height, spacing and number of the bottom surface of the microneedle array can be customized by changing the size of the mold. The microneedles 4 are arranged into a plurality of concentric circle structures, so that the administration is more uniform, and the administration effect is improved.
The third aspect of the invention provides a method for preparing a microneedle patch, comprising the following steps:
s1, mixing a microneedle skeleton material, plant growth hormone and a solvent to obtain a microneedle preparation;
s2, injecting the microneedle preparation into a mould, drying, and demoulding to obtain the microneedle array;
s3, assembling the microneedle array with a substrate and optional release paper to obtain the microneedle patch;
the microneedle skeleton material contains large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylate hyaluronic acid; the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:2-5:0.1-0.5.
The inventor finds that the microneedle skeleton material is limited to contain the three components according to the mass ratio and is mixed according to the mass ratio, and the prepared microneedle array can enable growth hormone to directly act on plant tissues and vascular systems through interaction of the three components, so that penetration and absorption of the plant tissues and vascular systems on the growth hormone are realized, a better delivery effect is achieved, and meanwhile, the traditional application of drug loss is avoided, and the application of a microneedle technology on plant drug delivery is realized. Further preferably, the mass ratio of the large molecular weight hyaluronic acid, the small molecular weight hyaluronic acid and the methacrylated hyaluronic acid is 1:2-4:0.2-0.5. And hyaluronic acid in the microneedle skeleton material can be dissolved in plant tissues, so that the prepared microneedle has the advantages of buffer administration and full administration, and meanwhile, the hyaluronic acid can accelerate the repair of wounds and play a role in curing the wounds.
Preferably, in step S1, the plant growth hormone may be auxin, gibberellin, cytokinin, abscisic acid, ethylene, etc., and preferably, the plant growth hormone is selected from at least one of auxin, gibberellin, and cytokinin. It has been found that the combination of the above plant growth hormone with the microneedle scaffold material can better allow the plant growth hormone in the microneedle preparation to act on plant growth. More preferably, the plant growth hormone is gibberellin.
Preferably, the mass ratio of the plant growth hormone, the microneedle skeleton material and the solvent is 1:10-300:100-3000. Under the condition of the mass ratio, the micro-needle array can be ensured to have better drug delivery effect on plants. From the viewpoint of further improving the administration effect of the micro-needle prepared from the microneedle preparation to plants, it is further preferable that the mass ratio of the plant growth hormone, the microneedle scaffold material, and the solvent is 1:100-200:1000-2000.
According to the present invention, the solvent is water or a buffer, and the buffer may be a conventionally used buffer such as a phosphate buffer, a carbonate buffer, or the like. The solvent is water or buffer solution, so that the microneedle skeleton material and the plant growth hormone have good dispersion effect, the plant growth hormone is uniformly released in an acting object, and the administration effect of the prepared microneedle array is improved. Further preferably, the solution is water.
Preferably, in step S2, the method of injecting the microneedle preparation into the mold is a horizontal centrifugation method or a negative pressure suction method, and the microneedle preparation can be spread on the mold more quickly and uniformly without generating small bubbles. Naturally air-dried at room temperature, the microneedle preparation can keep stability and can evaporate water as soon as possible.
Preferably, the microneedle array 3 includes a plurality of microneedles 4, and a plurality of the microneedles 4 are arranged into a plurality of concentric circle structures, and the microneedles 4 are arranged into a plurality of concentric circle structures, so that the administration is more uniform, the administration effect is improved, the waste is avoided, and the bottom surface diameter, the height, the spacing and the quantity of the microneedle array can be customized by changing the size of the die.
Preferably, in step S3, the method of assembling the microneedle array with a substrate and optionally a release paper includes: attaching the microneedle array to the substrate through a medical adhesive, attaching the release paper to the substrate through the medical adhesive, and penetrating the microneedle array out of an opening of the release paper.
Preferably, the microneedle patch prepared by the preparation method has two specifications, namely a single-piece package and a six-piece package, as shown in fig. 6.
Preferably, the mold is formed by copying a metal microneedle male mold and is in an inward inverted needle shape, the material is selected from PDMS, modified epoxy resin or silicic acid gel, and the preparation flow is shown in figure 3.
The fourth aspect of the invention provides the microneedle patch and application of the microneedle patch prepared by the method as a plant drug delivery carrier.
The application method of the microneedle patch serving as a plant drug delivery carrier to plants comprises the following steps:
(1) The preparation stage: before the microneedle patch is used, the surfaces (such as roots, stems and leaves) of the plants are cleaned, so that the plants are kept clean and dry, and meanwhile, the hands are kept dry after the hands are cleaned.
(2) And tearing a piece of plant growth hormone patch from the release paper, and placing the microneedle patch on the surface of a plant without touching the central microneedle array part by hands to ensure that the microneedle array part of the microneedle patch is completely covered on the surface of the plant.
(3) And (5) applying force to press, and tearing off the microneedle patch after applying for 2-12 hours.
In a relatively preferred embodiment of the present invention, there is provided a method for preparing a microneedle patch, comprising:
(1) The mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:2-4: and mixing 0.2-0.5 to obtain the microneedle skeleton material.
(2) The gibberellin, the microneedle skeleton material and water are mixed according to the mass ratio of 1:100-200: mixing 1000-2000 to obtain microneedle preparation;
(3) Filling the obtained microneedle preparation into a mould by a horizontal centrifugation method or a negative pressure air suction method, naturally air-drying at room temperature for 3-5h, and demoulding after drying to obtain a microneedle array;
(4) And adhering the microneedle array to a substrate, wherein the release paper is provided with an opening with the same size as the microneedle array, and the microneedle array penetrates out of the opening of the release paper and is attached to the substrate.
The microneedle patch prepared by the preparation method can be used for applying drugs to plants, so that the prepared microneedle array can enable growth hormone to directly act on plant tissues and vascular systems, so that penetration and absorption of the plant tissues and vascular systems on the growth hormone are realized, the microneedle preparation acts on plant drug delivery is realized, a better delivery effect can be achieved, and high-efficiency low-loss delivery of phytochemicals is realized.
The present invention will be described in detail by examples.
Unless otherwise specified, the following examples generally follow conventional conditions or recommended conditions by the reagent company; the reagents, consumables, etc. used in the examples described below are commercially available, and the process water is pure water, and the methods used are conventional in the art, unless otherwise specified.
In the following examples, small molecular weight hyaluronic acid and large molecular weight hyaluronic acid were purchased from Huaxi biotechnology Co., ltd. The methacrylated hyaluronic acid was purchased from Shanghai Ala Biotechnology Co., ltd. Gibberellin standard was purchased from Jiangxi Xinruifeng Biotechnology Co., ltd.
Example 1
Firstly, the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:3: mixing the materials according to the mass of 0.2 to obtain a microneedle skeleton material, dissolving 1g of the microneedle skeleton material in 200mL of water, adding 0.1g of gibberellin, and uniformly mixing to obtain the microneedle preparation.
And (3) injecting the prepared microneedle preparation into a mould (the preparation flow of the mould is shown in figure 3), putting the mould into a vacuum drying oven, opening a vacuum pump to uniformly and rapidly fill the microneedle preparation into inverted needle-shaped holes of the mould, naturally airing for 5 hours, and stripping the cured microneedle preparation from the mould by using tweezers to obtain the microneedle array.
And adhering the prepared microneedle array to a substrate, arranging holes with the same size as the microneedle array on release paper, and assembling the microneedle array to obtain the microneedle patch, wherein the microneedle array is suitable for penetrating through the holes of the release paper.
And carrying out SEM characterization on the prepared microneedle array, wherein the microneedle array is orderly arranged, an image of a microneedle body is obtained after the microneedle array is amplified, the needle body is complete and has no defect, the single microneedle is characterized, and the single microneedle is cone-shaped, as shown in A in FIG. 5.
Example 2
The microneedle patch was prepared according to the preparation method of example 1, except that the mass ratio of the large molecular weight hyaluronic acid, the small molecular weight hyaluronic acid and the methacrylated hyaluronic acid was 1:2: mixing the materials by mass of 0.5 to obtain a microneedle skeleton material, dissolving 1g of the microneedle skeleton material in 10mL of water, adding 0.1g of gibberellin, and uniformly mixing to obtain the microneedle preparation.
Example 3
The microneedle patch was prepared according to the preparation method of example 1, except that the mass ratio of the large molecular weight hyaluronic acid, the small molecular weight hyaluronic acid and the methacrylated hyaluronic acid was 1:5: and 0.1g of gibberellin is added and uniformly mixed to obtain a microneedle skeleton material, 1g of the microneedle skeleton material is dissolved in 300mL of water, and then 0.1g of gibberellin is added and uniformly mixed to obtain the microneedle preparation.
Comparative example 1
A microneedle patch was prepared as in example 1, except that the methacrylated hyaluronic acid was not added to the microneedle formulation.
Comparative example 2
A microneedle patch was prepared as in example 1, except that in the microneedle formulation, the methacrylated hyaluronic acid was replaced with silk fibroin.
Comparative example 3
A microneedle patch was prepared as in example 1, except that no large molecular weight hyaluronic acid and no small molecular weight hyaluronic acid were added to the microneedle preparation.
Comparative example 4
A microneedle patch was prepared as in example 1, except that no gibberellin was added to the microneedle preparation.
Test example 1
Selecting 8*5 Arabidopsis thaliana (respectively counted as A group, B group, C group, D group, E group, F group, G group and H group) with 30 days old as model plants, and spraying gibberellin-containing solution on Arabidopsis thaliana leaves of the A group by a fine mist sprayer (the spraying amount of gibberellin of each plant is 0.02G); the microneedle patches prepared in example 1 were equally divided into five groups, and the microneedle patches of each group were attached to the petioles of the plant leaves of the different plants of group B, respectively. The microneedle patches of examples 2-3 and comparative examples 1-4 were sequentially administered to arabidopsis thaliana in group C, group D, group E, group F, group G, and group H as in example 1 above. The three indices of the number of rosette leaves, the number of stem leaves and the bolting time were recorded, and the following results were obtained, as shown in table 1.
TABLE 1
| Group number | Experimental group | Number of rosette leaves (slices) | Number of stems and leaves (tablet) | Bolting time (Tian) |
| Group A | Traditional spray | 32±3 | 15±2 | 51±5 |
| Group B | Example 1 | 37±2 | 17±1 | 50±3 |
| Group C | Examples2 | 39±3 | 19±2 | 55±5 |
| Group D | Example 3 | 35±1 | 17±3 | 53±1 |
| Group E | Comparative example 1 | 30±3 | 17±4 | 40±3 |
| Group F | Comparative example 2 | 29±1 | 15±1 | 47±2 |
| Group G | Comparative example 3 | 26±2 | 10±3 | 39±5 |
| H group | Comparative example 4 | 20±3 | 5±1 | 31±3 |
As can be seen from Table 1, the plant growth rates of examples 1 to 3 were higher than those of the conventional spray group A, and the plant growth rates of the conventional spray group A were higher than those of comparative examples 1 to 4. The experimental result can be obtained, and the microneedle patch prepared by the microneedle preparation in the protection scope of the invention acts on plants and has better delivery effect.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, a number of simple variants of the technical solution of the invention are possible, including combinations of the individual technical features in any other suitable way, which simple variants and combinations should likewise be regarded as being disclosed by the invention, all falling within the scope of protection of the invention.
Claims (7)
1. A microneedle preparation, characterized in that the microneedle preparation comprises a microneedle skeleton material, a plant growth hormone and a solvent;
the microneedle skeleton material contains large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylate hyaluronic acid; the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:2-5:0.1-0.5;
the mass ratio of the plant growth hormone to the microneedle skeleton material to the solvent is 1:10-300:100-3000;
the plant growth hormone is at least one selected from auxin, gibberellin and cytokinin;
the solvent is water or buffer solution.
2. A microneedle patch, characterized in that it comprises a microneedle array (3) made of the microneedle preparation of claim 1 and a substrate (2), the microneedle array (3) being disposed on the substrate (2).
3. Microneedle patch according to claim 2, characterized in that the microneedle patch further comprises a release paper (1), the substrate (2) is attached to the release paper (1), and the release paper (1) is provided with openings, the microneedle array (3) being adapted to pass through the openings;
the substrate (2) is a hydrocolloid; the release paper (1) is glassine paper or polyethylene plastic film.
4. A microneedle patch according to claim 2 or 3, characterized in that the microneedle array (3) comprises a plurality of microneedles (4), a plurality of the microneedles (4) being arranged in a plurality of concentric circular structures.
5. A method of preparing the microneedle patch of claim 2, comprising the steps of:
s1, mixing a microneedle skeleton material, plant growth hormone and a solvent to obtain a microneedle preparation;
s2, injecting the microneedle preparation into a mould, drying, and demoulding to obtain a microneedle array (3);
s3, assembling the microneedle array (3) with a substrate (2) and a release paper (1) which is selectively added to obtain the microneedle patch;
the microneedle skeleton material contains large molecular weight hyaluronic acid, small molecular weight hyaluronic acid and methacrylate hyaluronic acid; the mass ratio of the large molecular weight hyaluronic acid to the small molecular weight hyaluronic acid to the methacrylate hyaluronic acid is 1:2-5:0.1-0.5;
the plant growth hormone is at least one selected from auxin, gibberellin and cytokinin;
the mass ratio of the plant growth hormone to the microneedle skeleton material to the solvent is 1:10-300:100-3000, wherein the solvent is water or buffer solution.
6. The method of manufacturing according to claim 5, wherein the microneedle array (3) comprises a plurality of microneedles (4), the plurality of microneedles (4) being arranged in a plurality of concentric circular structures;
in step S3, the method of assembling the microneedle array (3) with the substrate (2) and the optionally added release paper (1) comprises: attaching the microneedle array (3) to the substrate (2), attaching the release paper (1) to the substrate (2), and enabling the microneedle array (3) to penetrate out of the opening of the release paper (1).
7. Use of a microneedle patch according to any one of claims 2 to 4 or a microneedle patch prepared by a preparation method according to claim 5 or 6 as a carrier for plant drug delivery.
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| CN107184417A (en) * | 2017-03-31 | 2017-09-22 | 广州新济药业科技有限公司 | Soluble microneedle patch and preparation method thereof |
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| CN111904951A (en) * | 2020-08-15 | 2020-11-10 | 李媚 | Microneedle patch and preparation method thereof |
| WO2022142195A1 (en) * | 2020-12-31 | 2022-07-07 | 苏州悦肤达医疗科技有限公司 | Microneedle patch and preparation method therefor |
| KR20220141235A (en) * | 2021-04-12 | 2022-10-19 | 한미약품 주식회사 | Microneedle array and fabrication method thereof |
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| KR20160112975A (en) * | 2015-03-19 | 2016-09-28 | 라이온 가부시키가이샤 | Microneedle preparation and method for producing microneedle preparation |
| CN107184417A (en) * | 2017-03-31 | 2017-09-22 | 广州新济药业科技有限公司 | Soluble microneedle patch and preparation method thereof |
| CN108969880A (en) * | 2018-06-12 | 2018-12-11 | 上海白衣缘生物工程有限公司 | A kind of solubility micropin film and preparation method thereof |
| CN111904951A (en) * | 2020-08-15 | 2020-11-10 | 李媚 | Microneedle patch and preparation method thereof |
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| KR20220141235A (en) * | 2021-04-12 | 2022-10-19 | 한미약품 주식회사 | Microneedle array and fabrication method thereof |
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