CN116370400A - Phenylboronic acid modified chitosan-based microneedle patch and preparation method thereof - Google Patents
Phenylboronic acid modified chitosan-based microneedle patch and preparation method thereof Download PDFInfo
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- CN116370400A CN116370400A CN202310268802.XA CN202310268802A CN116370400A CN 116370400 A CN116370400 A CN 116370400A CN 202310268802 A CN202310268802 A CN 202310268802A CN 116370400 A CN116370400 A CN 116370400A
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- microneedle
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- chitosan
- mixed gel
- patch
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- HXITXNWTGFUOAU-UHFFFAOYSA-N phenylboronic acid Chemical compound OB(O)C1=CC=CC=C1 HXITXNWTGFUOAU-UHFFFAOYSA-N 0.000 title claims abstract description 73
- 229920001661 Chitosan Polymers 0.000 title claims abstract description 72
- 238000002360 preparation method Methods 0.000 title abstract description 10
- 239000000499 gel Substances 0.000 claims abstract description 98
- 239000000203 mixture Substances 0.000 claims abstract description 86
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims abstract description 74
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 71
- 239000008103 glucose Substances 0.000 claims abstract description 71
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- 102000004877 Insulin Human genes 0.000 claims abstract description 37
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 17
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 43
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 34
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- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 32
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 32
- 238000003756 stirring Methods 0.000 claims description 23
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 238000000502 dialysis Methods 0.000 claims description 21
- 239000002245 particle Substances 0.000 claims description 18
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 14
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- 239000008280 blood Substances 0.000 claims description 8
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- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 7
- KWNPRVWFJOSGMZ-UHFFFAOYSA-N 2-boronobenzoic acid Chemical compound OB(O)C1=CC=CC=C1C(O)=O KWNPRVWFJOSGMZ-UHFFFAOYSA-N 0.000 claims description 6
- SIAVMDKGVRXFAX-UHFFFAOYSA-N 4-carboxyphenylboronic acid Chemical compound OB(O)C1=CC=C(C(O)=O)C=C1 SIAVMDKGVRXFAX-UHFFFAOYSA-N 0.000 claims description 5
- 108010057186 Insulin Glargine Proteins 0.000 claims description 5
- COCFEDIXXNGUNL-RFKWWTKHSA-N Insulin glargine Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(=O)NCC(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)[C@@H](C)CC)[C@@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 COCFEDIXXNGUNL-RFKWWTKHSA-N 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 5
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- 238000004108 freeze drying Methods 0.000 claims description 5
- 229960002869 insulin glargine Drugs 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 238000005859 coupling reaction Methods 0.000 claims description 4
- 238000011049 filling Methods 0.000 claims description 4
- 238000002493 microarray Methods 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- AKTIXMMQDITKQU-UHFFFAOYSA-N 2-(4,5,5-trimethyl-2-phenyl-1,3,2-dioxaborolan-4-yl)acetic acid Chemical compound O1C(CC(O)=O)(C)C(C)(C)OB1C1=CC=CC=C1 AKTIXMMQDITKQU-UHFFFAOYSA-N 0.000 claims description 2
- WVKOYVWFDCAHKF-UHFFFAOYSA-N 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=CC(C(O)=O)=C1F WVKOYVWFDCAHKF-UHFFFAOYSA-N 0.000 claims description 2
- DBVFWZMQJQMJCB-UHFFFAOYSA-N 3-boronobenzoic acid Chemical compound OB(O)C1=CC=CC(C(O)=O)=C1 DBVFWZMQJQMJCB-UHFFFAOYSA-N 0.000 claims description 2
- CZDWJVSOQOMYGC-UHFFFAOYSA-N 4-borono-2-fluorobenzoic acid Chemical compound OB(O)C1=CC=C(C(O)=O)C(F)=C1 CZDWJVSOQOMYGC-UHFFFAOYSA-N 0.000 claims description 2
- 108010073961 Insulin Aspart Proteins 0.000 claims description 2
- 201000001421 hyperglycemia Diseases 0.000 claims description 2
- 229960004717 insulin aspart Drugs 0.000 claims description 2
- VOMXSOIBEJBQNF-UTTRGDHVSA-N novorapid Chemical compound C([C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CS)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CO)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CS)NC(=O)[C@H](CS)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O)C1=CC=C(O)C=C1.C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CS)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 VOMXSOIBEJBQNF-UTTRGDHVSA-N 0.000 claims description 2
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- 208000013016 Hypoglycemia Diseases 0.000 abstract description 5
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- 238000002347 injection Methods 0.000 abstract description 5
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- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
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- -1 phenylboronic acid modified glucose Chemical class 0.000 description 1
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Images
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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
- A61K9/0012—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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/28—Insulins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- 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
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
- A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
- A61M2037/0046—Solid microneedles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
- A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
- A61M2037/0053—Methods for producing microneedles
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Diabetes (AREA)
- Dermatology (AREA)
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- Hematology (AREA)
- Endocrinology (AREA)
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- Immunology (AREA)
- Medical Informatics (AREA)
- Anesthesiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Heart & Thoracic Surgery (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Emergency Medicine (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a phenylboronic acid modified chitosan-based microneedle patch and a preparation method thereof. The microneedle patch is prepared by attaching a mixed gel microneedle prepared by mixing a glucose response part, insulin and a polymer hydrogel to a back patch. The method comprises the following steps: dispersing glucose response part in dilute hydrochloric acid solution, adding insulin, adding polymer hydrogel, and mixing to obtain a precursor of microneedle mixed gel; injecting the mixture into a microneedle mould, placing the microneedle mould into a centrifuge for centrifugation, and drying the mixture to constant weight to obtain a mixed gel microneedle; and attaching the chitosan-based micro-needle patch to a back patch to prepare the chitosan-based micro-needle patch. The preparation method of the microneedle patch is simple and convenient, and the safety of raw materials is high. The method has the advantages of having sensitive glucose concentration responsiveness and a long-acting controllable release process, being painless and minimally invasive in the treatment process, effectively avoiding the risk of hypoglycemia, improving the bioavailability of the medicine, and reducing the treatment risk and side effect caused by subcutaneous insulin injection.
Description
Technical Field
The invention relates to a chitosan-based microneedle patch, in particular to a phenylboronic acid modified chitosan-based microneedle patch and a preparation method thereof.
Background
Diabetes is a disease that results in defective insulin secretion and blood glucose disorders due to impaired islet function. For type 1 and severe type 2 diabetics, the only treatment is the injection of exogenous insulin. The most common injection mode at present is syringe subcutaneous injection, and the treatment mode can bring great psychological treatment conflict and obvious injection pain to patients, and long-term subcutaneous injection can also cause irreversible damage to the skin of the patients. Thus, the microneedle can be used for delivering insulin transdermally, and thus, the microneedle becomes a potential better treatment means. The microneedles have size advantages that can avoid many of the disadvantages of subcutaneous injection. And through the design of the components and the structure in the micro needle, the insulin controllable release process of blood sugar concentration dependency can be realized. And simultaneously, the risk of hypoglycemia caused by excessive insulin treatment is avoided to a great extent.
At present, the micro-needles developed by researchers for treating diabetes are of two types. One type is a general microneedle, and the other type is a sugar-sensitive microneedle. Conventional microneedles can achieve painless, minimally invasive transdermal delivery of insulin, but the release process of insulin is uncontrolled and there may be a potential risk of oversherapy. The glucose-sensitive microneedle can realize the glucose-responsive release regulation and control process on the basis of painless minimally invasive. However, some glucose-responsive substances are biologically toxic or react with glucose to produce toxic byproducts, such as canavalin and glucose oxidase, which can cause unwanted toxic side effects to the human body. The phenylboronic acid group-based sugar-sensitive microneedle has good biological inertness. Can be excreted metabolically in the human body. Therefore, the glucose-sensitive microneedle can be used as a glucose responsive substance to prepare the glucose-sensitive microneedle, the controllability of the insulin release process is improved through the design of the phenylboronic acid-containing material, and the treatment safety of the glucose-sensitive microneedle is further improved through the combination of the glucose-sensitive microneedle and the material with good biocompatibility, so that the glucose-sensitive microneedle has important research significance for realizing clinical treatment as early as possible.
Disclosure of Invention
In order to solve the problems in the background art, the invention provides a phenylboronic acid modified chitosan-based microneedle patch and a preparation method thereof. The glucose-sensitive microneedle patch is prepared by taking phenylboronic acid modified chitosan particles as glucose responsive substances and PVA/PVP hydrogel as a microneedle needle material. The prepared microneedle body has sufficient skin penetration strength and toughness for preventing fracture, and can effectively load insulin and maintain pharmaceutical activity for a certain period of time. The method realizes the drug controllable release process with two-stage glucose responsiveness by a particle mixing gel mode.
The technical scheme adopted by the invention is as follows:
1. phenylboronic acid modified chitosan-based microneedle patch:
the chitosan-based microneedle patch is prepared by mixing a glucose response part, insulin and polymer hydrogel, and then attaching a mixed gel microneedle prepared by a microarray die to a back patch.
The glucose response part is chitosan grafted with phenylboronic acid through a coupling reaction, namely phenylboronic acid groups are modified on a chitosan chain segment through the coupling reaction, and the mole ratio of the phenylboronic acid groups in chitosan molecules is 1% -70%.
The insulin is insulin glargine or insulin aspart.
The polymer hydrogel is one or more of polyvinyl alcohol PVA and polyvinylpyrrolidone PVP, and when the polymer hydrogel is PVA/PVP mixed hydrogel of polyvinyl alcohol PVA and polyvinylpyrrolidone PVP, the mass ratio of the polyvinyl alcohol PVA to the polyvinylpyrrolidone PVP is 0/1, 1/2, 1/1, 2/1 or 1/0.
The chitosan has different molecular weights, and the molecular weight range is 3000-300000; the structural formula is specifically as follows:
the phenylboronic acid modified chitosan has the structural formula as follows:
-R is a phenylboronic acid group for modifying chitosan, one or more of the six groups shown below, the grafting ratio of phenylboronic acid groups being y/1:
2. a preparation method of phenylboronic acid modified chitosan-based microneedle patch comprises the following steps:
the method comprises the following steps:
1) The glucose responsive portion was prepared.
2) Uniformly dispersing the glucose response part in pure water, then adding a dilute hydrochloric acid solution until the glucose response part is completely dissolved, then adding insulin, uniformly stirring and mixing, finally adding a polymer hydrogel, uniformly stirring and mixing, sealing and standing to obtain a precursor of the microneedle mixed gel.
3) Injecting the precursor of the microneedle mixed gel into a microneedle mould and filling the microneedle mould, placing the microneedle mould in a centrifuge for centrifugal treatment, scraping the surface of the microneedle mould after the centrifugal treatment is finished, removing redundant gel, and placing the microneedle mould in a constant-temperature and constant-humidity dryer to dry the precursor of the microneedle mixed gel to constant weight, so as to obtain the phenylboronic acid modified chitosan-based mixed gel microneedle carrying insulin; and attaching the mixed gel microneedle to a back patch to prepare the chitosan-based microneedle patch.
In the step 1), the glucose response part is prepared specifically as follows:
1.1 Chitosan particles were dissolved in pure water, and then diluted hydrochloric acid solution was added until complete dissolution, to obtain a first mixture a.
1.2 Dissolving carboxyphenylboronic acid, 1 (3 dimethylaminopropyl) 3 ethylcarbodiimide hydrochloride EDC and N hydroxysuccinimide NHS in pure water or an organic solution until complete dissolution, to give a second mixture B.
1.3 Slowly adding the first mixture A into the second mixture B, uniformly stirring, adding sodium hydroxide solution at normal temperature, and obtaining a third mixture C after the reaction is completed.
1.4 Pouring the third mixture C into a dialysis bag, dialyzing in pure water, and placing the dialysis bag into a water area with medium temperature for rotary evaporation after the dialysis is completed to obtain the evaporated third mixture C.
1.5 Freeze-drying the evaporated third mixture C to completely remove the solution, thereby obtaining the final product phenylboronic acid modified chitosan powder, namely a glucose response part.
In the step 1.1), chitosan particles are dissolved in 0.5-100mL of pure water, and then 40-100mL of diluted hydrochloric acid solution with ph=1 is added until complete dissolution, thus obtaining a first mixture a.
In the step 1.2), 1-5g of carboxyphenylboronic acid, 1-5.5g of 1 (3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride EDC and 0.5-3.5g of N-hydroxysuccinimide NHS are dissolved in 30-100mL of pure water or an organic solution until the solution is completely dissolved, so as to obtain a second mixture B.
In the step 1.2), the carboxyphenylboronic acid is one or more of 3-carboxyphenylboronic acid, 4-carboxyphenylboronic acid, carboxyphenylboronic acid pinacol ester, 4-carboxy 3-fluorobenzeneboronic acid and carboxyfluorobenzeneboronic acid pinacol ester.
In the step 1.3), the first mixture A is slowly added into the second mixture B and uniformly stirred, in the process of adding the first mixture A, the magnetons keep the rotating speed of 500-700rpm and uniformly stirred for 30min, then 40mL of sodium hydroxide solution with the pH value of=1 is added, the pH value is regulated to 6-7, and the third mixture C is obtained after the reaction is carried out for 3-48h at normal temperature.
In the step 1.4), pouring the third mixture C into a dialysis bag, dialyzing in 5L of pure water for 3-5 days, changing water once every 12 hours, placing the dialysis bag into a water area with the temperature of 40-55 ℃ for rotary evaporation after the dialysis is finished, and reducing the rotary evaporation of the solvent in the third mixture C to 20-35mL to obtain the evaporated third mixture C.
In the step 2), 0.5-50mg of glucose response part is uniformly dispersed in 0.5-100mL of pure water, added into 0.1-5mL of dilute hydrochloric acid solution with pH=6 until the glucose response part is completely dissolved, and then added with 0.1-10mg of insulin, and uniformly stirred and mixed to obtain a fourth mixture D; dissolving 10-5000mg of polymer hydrogel in 0.5-100mL of pure water, and uniformly stirring and mixing to obtain a fifth mixture E; uniformly mixing the fourth mixture D and the fifth mixture E, stirring for 0.5-2h at a rotating speed of 25-100rpm, and hermetically standing for 3-24h in a shading environment to obtain a precursor of the microneedle mixed gel; the mass fraction of the dilute hydrochloric acid solution of the glucose response part is 20% -80%; the mass fraction of the polymer hydrogel is 15% -45%;
in the step 3), 50-500mg of precursor of the microneedle mixed gel is injected into a microneedle mould and filled, the microneedle mould is placed in a centrifuge for centrifugal treatment for 5-60min at the centrifugal rotation speed of 1000-10000rpm, the microneedle mould is placed in a dryer with the relative humidity of 40% at 25 ℃ after the centrifugal treatment is finished, the precursor of the microneedle mixed gel is dried for 12-24h to constant weight of 2-100mg and is stored in a shading mode for 24h, and the phenylboronic acid modified chitosan-based mixed gel microneedle carrying insulin is obtained.
In the step 3), the mixed gel microneedle is composed of a mixed gel sheet body and a plurality of mixed gel needle bodies, the mixed gel needle body arrays are uniformly distributed on one side face of the mixed gel sheet body, and the other side face of the mixed gel sheet body is attached to the back patch.
The length of each mixed gel needle body is 0.1-2.0mm, and the bottom area is 0.01-4mm 2 The array density of each mixed gel needle body on the back patch is 16-1000 needles per patch, and the array specification can be 5×5-100×100; the area of the back patch is 0.01-4cm 2 The thickness is 0.1-0.5cm.
3. An application method of phenylboronic acid modified chitosan-based microneedle patch comprises the following steps:
the side, on which the mixed gel needle body is attached, of the chitosan-based microneedle is attached to the skin of a human body suffering from hyperglycemia through the back patch, so that each mixed gel needle body in the mixed gel microneedle pierces the skin and the glucose concentration-dependent insulin in the mixed gel microneedle is released into the human body in a long-acting and controllable manner, and the chitosan-based microneedle is used for adjusting the blood glucose concentration.
The beneficial effects of the invention are as follows:
1. the preparation process of the microneedle patch is simple and convenient, the raw materials are easy to prepare, and the used raw materials have good biological safety; has larger insulin loading capacity, and chitosan and PVA/PVP in the main material can be metabolized and discharged in a short period after human circulation, and no toxic or side effect is generated in the body.
2. The microneedle body material of the microneedle patch has good skin penetration capability, safe and effective skin penetration effect and high-efficiency insulin delivery process, and good toughness, and can prevent breakage and loss in transportation and treatment processes.
3. The microneedle patch provided by the invention has good glucose concentration responsiveness; and the change of the glucose concentration can be rapidly identified through the two-stage controlled release process, and the quick response and long-acting insulin release process of the change of the glucose concentration can be realized through regulating and controlling the release rate of insulin through the PVA/PVP gel network. The risk of hypoglycemia which may occur due to sudden insulin release is effectively avoided. Meanwhile, the waste of medicines is avoided to a certain extent, and the bioavailability is improved.
In a word, the preparation method of the microneedle provided by the invention is simple and convenient, and the safety of raw materials is high. Has sensitive glucose concentration response and long-acting controllable release process. The treatment process is painless and minimally invasive, the risk of hypoglycemia is effectively avoided, and the bioavailability of the medicine is improved. And through treatment of phenylboronic acid modified chitosan-based microneedle patches, patient compliance can be improved to a great extent, and treatment risks and side effects caused by subcutaneous insulin injection can be reduced.
Drawings
FIG. 1 is a flow chart showing the preparation of a microneedle patch according to example 1 of the present invention;
FIG. 2 (a) is a microarray Scanning Electron Microscope (SEM) image of the microneedle patch of example 1 of the present invention;
FIG. 2 (b) is a Scanning Electron Microscope (SEM) image of the micro-array structure of the microneedle patch of example 1 of the present invention;
FIG. 3 (a) is an in vitro glucose-sensitive release profile of the microneedle patch microneedle of example 2 of the present invention;
FIG. 3 (b) is an in vitro glucose-sensitive release scanning electron microscope image of the microneedle patch microneedle of example 2 of the present invention;
FIG. 3 (c) is an in vitro glucose-sensitive release partial-magnifying Scanning Electron Microscope (SEM) of the microneedle patch of example 2 of the present invention;
FIG. 4 (a) is a graph showing the results of mechanical strength test of the microneedle patch of example 2 of the present invention;
FIG. 4 (b) is a mechanical strength test Scanning Electron Microscope (SEM) of the microneedle patch of example 2 of the present invention;
FIG. 5 (a) is a graph showing the in vivo blood glucose level lowering test of the microneedle patch of example 3 of the present invention;
FIG. 5 (b) is a schematic diagram of SD rats treated with the microneedle patch of example 3 of the present invention;
fig. 5 (c) is a schematic diagram of the stained area after microneedle therapy according to example 3 of the present invention.
Detailed Description
The present invention will be described in more detail with reference to the drawings and examples, but the invention is not limited thereto, and it will be apparent to those skilled in the art that modifications and variations can be made without departing from the principle of the present invention, and these modifications and variations are considered to be within the scope of the present invention. What is not described in detail in this specification is prior art known to those skilled in the art.
Specific embodiments of the invention are as follows:
example 1:
preparing a glucose responsive portion, dissolving 0.5033g of chitosan particles having a molecular weight of 3000 in 30mL of pure water, and then adding 40mL of a diluted hydrochloric acid solution having ph=1 to complete dissolution, to obtain a first mixture a; 1.1037g of 4-carboxyphenylboronic acid, 1.1501g of 1 (3 dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride EDC and 0.6913g of N-hydroxysuccinimide NHS were dissolved in 30mL of dimethyl sulfoxide to complete dissolution, giving a second mixture B; slowly adding the first mixture A into the second mixture B and uniformly stirring, wherein in the process of adding the first mixture A, the magneton keeps a rotating speed of 500rpm and uniformly stirring for 30min, then adding 40mL of sodium hydroxide solution with pH=1, adjusting the pH value to 7, and reacting at 25 ℃ for 12h to obtain a third mixture C; in the step 1.4), pouring the third mixture C into a dialysis bag, dialyzing in 5L of pure water for 3 days, changing water once every 12 hours, placing the dialysis bag into a water area with the temperature of 40 ℃ for rotary evaporation after the dialysis is finished, and reducing the rotary evaporation of the solvent in the third mixture C to 30mL to obtain an evaporated third mixture C; and (3) freeze-drying the evaporated third mixture C to completely remove the solution, thereby obtaining the final product phenylboronic acid modified chitosan powder, namely a glucose response part, wherein the mole ratio of phenylboronic acid groups of the glucose response part in chitosan molecules is 39%.
Uniformly dispersing 20mg of glucose responsive part in 1mL of pure water, adding into 0.2mL of dilute hydrochloric acid solution with pH=6 to completely dissolve, then adding 5.5mg of insulin glargine, and uniformly stirring and mixing to obtain a fourth mixture D;880mg of PVA/PVP mixed hydrogel formed by polyvinyl alcohol PVA and 880mg of polyvinylpyrrolidone PVP are dissolved in 10mL of pure water and uniformly stirred and mixed to obtain a fifth mixture E; uniformly mixing the fourth mixture D and the fifth mixture E, stirring for 1h at a rotating speed of 50rpm, and hermetically standing for 12h in a shading environment to obtain a precursor of the microneedle mixed gel.
As shown in fig. 1, a microneedle fabrication process is shown. 200mg of precursor of the microneedle mixed gel is injected into a microneedle mould and filled, the microneedle mould is placed in a centrifuge for centrifugal treatment for 30min at a centrifugal speed of 4500rpm, the microneedle mould is placed in a dryer with a relative humidity of 40% at 25 ℃ after the centrifugal treatment is finished, the precursor of the microneedle mixed gel is dried for 24h to constant weight of 15mg and stored in a shading mode, and the phenylboronic acid modified chitosan-based mixed gel microneedle carrying insulin is obtained. And attaching the mixed gel microneedle to a back patch to prepare the chitosan-based microneedle patch.
As shown in FIGS. 2 (a) and (b), for the prepared microneedles, a scanning electron microscope is usedAnd shooting a orderly arranged microneedle array and a full and complete microneedle structure. The mixed gel microneedle consists of a mixed gel sheet body and a plurality of mixed gel needle bodies, wherein each mixed gel needle body array is uniformly arranged on one side surface of the mixed gel sheet body, and the other side surface of the mixed gel sheet body is attached to the back patch; the length of each mixed gel needle body is 0.566mm, the width of the bottom of the needle body is 0.196mm, the distance between the needle points is 0.507mm, and the array specification of each mixed gel needle body on the back patch is 15 multiplied by 15; the area of the back patch is 1cm 2 The thickness was 0.5cm.
Example 2:
preparing a glucose responsive portion, dissolving 0.5031g of chitosan particles having a molecular weight of 300000 in 30mL of pure water, and then adding 40mL of a diluted hydrochloric acid solution having ph=1 to complete dissolution, to obtain a first mixture a; 1.1031g of 4-carboxyphenylboronic acid, 1.1502g of 1 (3 dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride EDC and 0.6901g of N-hydroxysuccinimide NHS were dissolved in 30mL of dimethyl sulfoxide to complete dissolution, giving a second mixture B; slowly adding the first mixture A into the second mixture B and uniformly stirring, wherein in the process of adding the first mixture A, the magneton keeps the rotating speed of 600rpm, uniformly stirring for 30min, then adding 40mL of sodium hydroxide solution with pH=1, adjusting the pH value to 7, and reacting for 12h at normal temperature to obtain a third mixture C; in the step 1.4), pouring the third mixture C into a dialysis bag, dialyzing in 5L of pure water for 3 days, changing water once every 12 hours, placing the dialysis bag into a water area with the temperature of 40 ℃ for rotary evaporation after the dialysis is finished, and reducing the rotary evaporation of the solvent in the third mixture C to 20mL to obtain an evaporated third mixture C; and (3) freeze-drying the evaporated third mixture C to completely remove the solution, thereby obtaining the final product phenylboronic acid modified chitosan powder, namely a glucose response part, wherein the mole ratio of phenylboronic acid groups of the glucose response part in chitosan molecules is 45%.
Uniformly dispersing 20mg of glucose response part in 1mL of pure water, then adding 0.2mL of dilute hydrochloric acid solution with pH=6 for auxiliary dispersion, then adding 5.5mg of insulin glargine, and uniformly stirring and mixing to obtain a fourth mixture D;1320mg of PVA/PVP mixed hydrogel formed by polyvinyl alcohol PVA and 1320mg of polyvinylpyrrolidone PVP is dissolved in 10mL of pure water and stirred uniformly to obtain a fifth mixture E; uniformly mixing the fourth mixture D and the fifth mixture E, stirring for 1h at a rotating speed of 50rpm, and hermetically standing for 12h in a shading environment to obtain a precursor of the microneedle mixed gel.
And (3) injecting 200mg of precursor of the needle mixed gel into a microneedle mould and filling the microneedle mould, putting the microneedle mould into a centrifuge for centrifugal treatment for 30min at a centrifugal speed of 4000rpm, putting the microneedle mould into a dryer with a relative humidity of 40% at 25 ℃ after the centrifugal treatment is finished, drying the precursor of the microneedle mixed gel for 24h to constant weight of 20mg, and carrying out shading preservation to obtain the insulin-loaded phenylboronic acid modified chitosan-based mixed gel microneedle. And attaching the mixed gel microneedle to a back patch to prepare the chitosan-based microneedle patch.
The mixed gel microneedle consists of a mixed gel sheet body and a plurality of mixed gel needle bodies, wherein each mixed gel needle body array is uniformly arranged on one side surface of the mixed gel sheet body, and the other side surface of the mixed gel sheet body is attached to the back patch; the length of each mixed gel needle is 567 mu m, the width of the bottom of the needle is 0.9mm, and the array specification of each mixed gel needle on the back patch is 15×15; the area of the back patch was 0.81cm 2 The thickness was 0.38cm.
As shown in FIG. 3, the prepared microneedles were subjected to in vitro release tests in phosphate buffers of different glucose concentrations. The amount of insulin released in the microneedles increased with increasing glucose concentration at 37 ℃. The glucose sensitivity performance and the controllable release characteristic of insulin of the micro needle are respectively reflected in glucose concentration solutions of 100mg/dL, 200mg/dL and 400 mg/dL. The CS 5-39% + PP sample in FIG. 3 (a) is a mixed gel of chitosan particles with 39% phenylboronic acid grafting with PVA/PVP. Fig. 3 (b) and 3 (c) are the distribution and size of the mixed gel in the 100mg/dL and 400mg/dL environments. Fig. 4 (a) shows the mechanical compression test performance of the microneedle, and the mechanical strength of the microneedle enough to penetrate the surface layer of the skin was measured by compressing the microneedle in the vertical direction. And during the compression process, the micro-needle can maintain good toughness and needle integrity. The PP is a microneedle prepared from PVA/PVP gel, the CS5-PP is a microneedle prepared from phenylboronic acid grafted chitosan particles and PVA/PVP mixed gel, and the CS 5-INS-PP is a microneedle prepared from phenylboronic acid grafted chitosan particles and PVA/PVP mixed gel loaded with insulin. Fig. 4 (b) shows the compressed shape of the microneedle.
Example 3:
preparing a glucose responsive portion, dissolving 0.5035g of chitosan particles having a molecular weight of 300000 in 30mL of pure water, and then adding 40mL of a diluted hydrochloric acid solution having ph=1 to complete dissolution, to obtain a first mixture a; 1.1025g of 4-carboxyphenylboronic acid, 1.1499g of 1 (3 dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride EDC and 0.6909g of N-hydroxysuccinimide NHS were dissolved in 30mL of dimethyl sulfoxide to complete dissolution, giving a second mixture B; slowly adding the first mixture A into the second mixture B and uniformly stirring, wherein in the process of adding the first mixture A, the magneton keeps a rotating speed of 700rpm and uniformly stirring for 30min, then 40mL of sodium hydroxide solution with pH=1 is added, the pH value is regulated to 7, and a third mixture C is obtained after reaction for 12h at normal temperature; in the step 1.4), pouring the third mixture C into a dialysis bag, dialyzing in 5L of pure water for 3 days, changing water once every 12 hours, placing the dialysis bag into a water area with the temperature of 40 ℃ for rotary evaporation after the dialysis is finished, and reducing the rotary evaporation of the solvent in the third mixture C to 20mL to obtain an evaporated third mixture C; and (3) freeze-drying the evaporated third mixture C to completely remove the solution, thereby obtaining the final product phenylboronic acid modified chitosan powder, namely a glucose response part, wherein the molar ratio of phenylboronic acid groups of the glucose response part in chitosan molecules is 32%.
Uniformly dispersing 20mg of glucose response part in 1mL of pure water, then adding 0.2mL of dilute hydrochloric acid solution with pH=6 for auxiliary dispersion, then adding 5.5mg of insulin glargine, and uniformly stirring and mixing to obtain a fourth mixture D; dissolving PVA/PVP mixed hydrogel formed by 1100mg of polyvinyl alcohol PVA and 1100mg of polyvinylpyrrolidone PVP in 10mL of pure water, and uniformly stirring and mixing to obtain a fifth mixture E; uniformly mixing the fourth mixture D and the fifth mixture E, stirring for 1h at a rotating speed of 50rpm, and hermetically standing for 12h in a shading environment to obtain a precursor of the microneedle mixed gel.
And (3) injecting 200mg of precursor of the microneedle mixed gel into a microneedle mould and filling the microneedle mould, putting the microneedle mould into a centrifuge for centrifugation at 5000rpm for 30min, putting the microneedle mould into a dryer with the relative humidity of 40% at 25 ℃ after the centrifugation is finished, drying the precursor of the microneedle mixed gel for 24h to constant weight of 16mg, and shading and preserving the precursor for 24h to obtain the insulin-loaded phenylboronic acid modified chitosan-based mixed gel microneedle. And attaching the mixed gel microneedle to a back patch to prepare the chitosan-based microneedle patch.
The mixed gel microneedle consists of a mixed gel sheet body and a plurality of mixed gel needle bodies, wherein each mixed gel needle body array is uniformly arranged on one side surface of the mixed gel sheet body, and the other side surface of the mixed gel sheet body is attached to the back patch; the length of each mixed gel needle body is 526mm, the width of the bottom of the needle body is 0.89mm, and the array specification of each mixed gel needle body on the back patch can be 15×15; the area of the back patch was 0.79cm 2 The thickness was 0.39cm.
As shown in fig. 5 (a), a microneedle patch was applied to SD rats, and a blood glucose control in vivo test was performed. In contrast to untreated diabetic rats, the microneedle-treated group was effective in lowering blood glucose and maintaining its blood glucose level in the normal range for at least 8 hours. Compared with a diabetic rat treated by subcutaneous injection, the microneedle treatment group can effectively avoid hypoglycemia caused by excessive release of insulin. Diabetes is Diabetes, MN1-CS5PBA-PP is microneedle therapy, MN2-CS5PBA-PP is microneedle therapy, MN3-CS5-PP is microneedle therapy, SI-INS is subcutaneous therapy, and health is Healthy. Fig. 5 (b) is a microneedle-treated SD rat; fig. 5 (c) shows the stained area after microneedle treatment.
The phenylboronic acid modified glucose responsive mixed gel microneedle prepared by the invention has rapid glucose concentration response capability and long-acting insulin controllable release process. The microneedle patch can load insulin of more than or equal to 10IU, and at least can meet the medicament demand of common diabetics for 24 hours. Through adjusting the PVA/PVP content of the microneedle needle material, the microneedle has enough mechanical strength for puncturing the skin, and simultaneously maintains certain toughness, so that the breakage and loss of the microneedle in the treatment process are prevented. When the microneedle penetrates the skin and reaches the dermis layer, the phenylboronic acid modified chitosan particles can be specifically combined with glucose molecules, so that the water solubility of the chitosan particles is changed, the particles swell, insulin coated inside is released, and the release process of the first stage is completed. The PVA component in the PVA/PVP gel outside the particle can then form a borate bond with the phenylboronic acid groups on the chitosan segment. Thus, an external gel network with dynamic covalent bonds is constructed, and the release process of insulin is delayed. When the concentration of glucose in the interstitial fluid is further improved, glucose molecules can open a dynamic gel network formed by PVA and phenylboronic acid groups, so that insulin is continuously released, and the release process of the second stage is completed. Both release phases are reversible and have a strong dependence on the external glucose concentration, thus having good controllability on the insulin release process.
Claims (10)
1. A phenylboronic acid modified chitosan-based microneedle patch is characterized in that: the chitosan-based microneedle patch is prepared by mixing a glucose response part, insulin and polymer hydrogel and then attaching a mixed gel microneedle prepared by a microarray die to a back patch.
2. The phenylboronic acid modified chitosan-based microneedle patch of claim 1, wherein the patch is characterized in that: the glucose response part is chitosan grafted with phenylboronic acid through a coupling reaction, namely phenylboronic acid groups are modified on a chitosan chain segment through the coupling reaction, and the molar ratio of the phenylboronic acid groups in chitosan molecules is 1% -70%;
the insulin is insulin glargine or insulin aspart;
the polymer hydrogel is one or more of polyvinyl alcohol PVA and polyvinylpyrrolidone PVP, and when the polymer hydrogel is PVA/PVP mixed hydrogel of polyvinyl alcohol PVA and polyvinylpyrrolidone PVP, the mass ratio of the polyvinyl alcohol PVA to the polyvinylpyrrolidone PVP is 0/1, 1/2, 1/1, 2/1 or 1/0.
3. The method for preparing the chitosan-based microneedle patch according to any one of claims 1 to 2, which is characterized by comprising the following steps: the method comprises the following steps:
1) Preparing a glucose responsive moiety;
2) Uniformly dispersing the glucose response part in pure water, then adding a dilute hydrochloric acid solution until the glucose response part is completely dissolved, then adding insulin, uniformly stirring and mixing, finally adding a polymer hydrogel, uniformly stirring and mixing, sealing and standing to obtain a precursor of the microneedle mixed gel;
3) Injecting the precursor of the microneedle mixed gel into a microneedle mould and filling the microneedle mould, placing the microneedle mould into a centrifuge for centrifugal treatment, and placing the microneedle mould into a dryer after the centrifugal treatment is finished so that the precursor of the microneedle mixed gel is dried to constant weight to obtain the mixed gel microneedle; and attaching the mixed gel microneedle to a back patch to prepare the chitosan-based microneedle patch.
4. A method for preparing a chitosan-based microneedle patch according to claim 3, wherein: in the step 1), the glucose response part is prepared specifically as follows:
1.1 Dissolving chitosan particles in pure water, and then adding a dilute hydrochloric acid solution until the chitosan particles are completely dissolved to obtain a first mixture A;
1.2 Dissolving carboxyphenylboronic acid, 1 (3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride EDC and N-hydroxysuccinimide NHS in pure water or an organic solution until the solution is completely dissolved to obtain a second mixture B;
1.3 Adding the first mixture A into the second mixture B, uniformly stirring, adding sodium hydroxide solution at normal temperature, and obtaining a third mixture C after the reaction is completed;
1.4 Pouring the third mixture C into a dialysis bag, dialyzing in pure water, and placing the dialysis bag into a water area with medium temperature for rotary evaporation after the dialysis is completed to obtain an evaporated third mixture C;
1.5 Freeze-drying the evaporated third mixture C to completely remove the solution, thereby obtaining the final product phenylboronic acid modified chitosan powder, namely a glucose response part.
5. The method for preparing the chitosan-based microneedle patch according to claim 4, wherein the method comprises the following steps: in the step 1.1), chitosan particles are dissolved in 0.5-100mL of pure water, and then 40-100mL of dilute hydrochloric acid solution with pH=1 is added until the chitosan particles are completely dissolved, so as to obtain a first mixture A;
in the step 1.2), 1-5g of carboxyphenylboronic acid, 1-5.5g of 1 (3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride EDC and 0.5-3.5g of N-hydroxysuccinimide NHS are dissolved in 30-100mL of pure water or an organic solution until the solution is completely dissolved, so as to obtain a second mixture B.
6. The method for preparing the chitosan-based microneedle patch according to claim 4, wherein the method comprises the following steps: in the step 1.2), the carboxyphenylboronic acid is one or more of 3-carboxyphenylboronic acid, 4-carboxyphenylboronic acid, carboxyphenylboronic acid pinacol ester, 4-carboxy 3-fluorobenzeneboronic acid and carboxyfluorobenzeneboronic acid pinacol ester.
7. The method for preparing the chitosan-based microneedle patch according to claim 4, wherein the method comprises the following steps: in the step 1.3), the first mixture A is added into the second mixture B and uniformly stirred, in the process of adding the first mixture A, the magneton is kept at a rotating speed of 500-700rpm and uniformly stirred for 30min, then 40mL of sodium hydroxide solution with pH value of 1 is added, the pH value is regulated to 6-7, and the third mixture C is obtained after the reaction is carried out for 3-48h at normal temperature;
in the step 1.4), pouring the third mixture C into a dialysis bag, dialyzing in 5L of pure water for 3-5 days, changing water once every 12 hours, placing the dialysis bag into a water area with the temperature of 40-55 ℃ for rotary evaporation after the dialysis is finished, and reducing the rotary evaporation of the solvent in the third mixture C to 20-35mL to obtain the evaporated third mixture C.
8. The method for preparing the chitosan-based microneedle patch according to claim 2, wherein the method comprises the following steps: in the step 2), 0.5-50mg of glucose response part is uniformly dispersed in 0.5-100mL of pure water, added into 0.1-5mL of dilute hydrochloric acid solution with pH=6 until the glucose response part is completely dissolved, and then added with 0.1-10mg of insulin, and uniformly stirred and mixed to obtain a fourth mixture D; dissolving 10-5000mg of polymer hydrogel in 0.5-100mL of pure water, and uniformly stirring and mixing to obtain a fifth mixture E; uniformly mixing the fourth mixture D and the fifth mixture E, stirring for 0.5-2h at a rotating speed of 25-100rpm, and hermetically standing for 3-24h in a shading environment to obtain a precursor of the microneedle mixed gel; the mass fraction of the dilute hydrochloric acid solution of the glucose response part is 20% -80%; the mass fraction of the polymer hydrogel is 15% -45%;
in the step 3), the precursor of the microneedle mixed gel is injected into a microneedle mould and filled, the microneedle mould is placed in a centrifuge for centrifugal treatment for 5-60min at the centrifugal speed of 1000-10000rpm, the microneedle mould is placed in a dryer after the centrifugal treatment is finished, the precursor of the microneedle mixed gel is dried for 12-24h to constant weight of 2-100mg and is stored in a shading mode for 24h, and the mixed gel microneedle is obtained.
9. The method for preparing the chitosan-based microneedle patch according to claim 2, wherein the method comprises the following steps: in the step 3), the mixed gel microneedle is composed of a mixed gel sheet body and a plurality of mixed gel needle bodies, wherein each mixed gel needle body array is uniformly arranged on one side surface of the mixed gel sheet body, and the other side surface of the mixed gel sheet body is attached to the back patch;
the length of each mixed gel needle body is 0.1-2.0mm, and the bottom area is 0.01-4mm 2 The array density of each mixed gel needle body on the back patch is 16-1000 needles per patch; the area of the back patch is 0.01-4cm 2 The thickness is 0.1-0.5cm.
10. A chitosan-based microneedle patch according to any one of claims 1 to 2 or a method of using the chitosan-based microneedle patch according to any one of claims 3 to 9, characterized in that: the side, on which the mixed gel needle body is attached, of the chitosan-based microneedle is attached to the skin of a human body suffering from hyperglycemia through the back patch, so that each mixed gel needle body in the mixed gel microneedle pierces the skin and releases insulin in the mixed gel microneedle into the human body for adjusting the blood sugar concentration.
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