CN114632145B - Insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and preparation method thereof - Google Patents
Insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and preparation method thereof Download PDFInfo
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- 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 title claims abstract description 212
- HXITXNWTGFUOAU-UHFFFAOYSA-N phenylboronic acid Chemical compound OB(O)C1=CC=CC=C1 HXITXNWTGFUOAU-UHFFFAOYSA-N 0.000 title claims abstract description 173
- 102000004877 Insulin Human genes 0.000 title claims abstract description 103
- 108090001061 Insulin Proteins 0.000 title claims abstract description 103
- 229940125396 insulin Drugs 0.000 title claims abstract description 102
- 108010039918 Polylysine Proteins 0.000 title claims abstract description 98
- 235000014113 dietary fatty acids Nutrition 0.000 title claims abstract description 80
- 229930195729 fatty acid Natural products 0.000 title claims abstract description 80
- 239000000194 fatty acid Substances 0.000 title claims abstract description 80
- 150000004665 fatty acids Chemical class 0.000 title claims abstract description 80
- 239000002245 particle Substances 0.000 title claims abstract description 66
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 239000000243 solution Substances 0.000 claims abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims abstract description 4
- 238000010254 subcutaneous injection Methods 0.000 claims abstract description 4
- 239000007929 subcutaneous injection Substances 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims description 81
- 238000000502 dialysis Methods 0.000 claims description 76
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 53
- 238000003756 stirring Methods 0.000 claims description 43
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 22
- 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 20
- 239000002904 solvent Substances 0.000 claims description 15
- 229920000642 polymer Polymers 0.000 claims description 14
- 238000004108 freeze drying Methods 0.000 claims description 12
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 10
- GHVNFZFCNZKVNT-UHFFFAOYSA-N decanoic acid Chemical compound CCCCCCCCCC(O)=O GHVNFZFCNZKVNT-UHFFFAOYSA-N 0.000 claims description 10
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 7
- 101000976075 Homo sapiens Insulin Proteins 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- PBGKTOXHQIOBKM-FHFVDXKLSA-N insulin (human) 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)=O)C(=O)N[C@@H](CC(N)=O)C(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 PBGKTOXHQIOBKM-FHFVDXKLSA-N 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 101000993800 Sus scrofa Insulin Proteins 0.000 claims description 3
- 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 abstract description 26
- 239000008103 glucose Substances 0.000 abstract description 26
- 239000008280 blood Substances 0.000 abstract description 16
- 210000004369 blood Anatomy 0.000 abstract description 16
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 16
- 208000013016 Hypoglycemia Diseases 0.000 abstract description 15
- 208000024891 symptom Diseases 0.000 abstract description 8
- 201000001421 hyperglycemia Diseases 0.000 abstract description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 239000013505 freshwater Substances 0.000 description 9
- 239000000047 product Substances 0.000 description 8
- 206010012601 diabetes mellitus Diseases 0.000 description 7
- 108010005991 Pork Regular Insulin Proteins 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 230000003914 insulin secretion Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 4
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 4
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 4
- 238000009825 accumulation Methods 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 229960001701 chloroform Drugs 0.000 description 3
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 239000005639 Lauric acid Substances 0.000 description 2
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 2
- 235000021314 Palmitic acid Nutrition 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- -1 carboxyl modified phenylboronic acid Chemical class 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 230000001771 impaired effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000000693 micelle Substances 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 229920002085 Dialdehyde starch Polymers 0.000 description 1
- 102100023915 Insulin Human genes 0.000 description 1
- 108010057186 Insulin Glargine Proteins 0.000 description 1
- 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 description 1
- 108010092217 Long-Acting Insulin Proteins 0.000 description 1
- 102000016261 Long-Acting Insulin Human genes 0.000 description 1
- 229940100066 Long-acting insulin Drugs 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 230000003095 anti-phagocytic effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000009748 deglutition Effects 0.000 description 1
- 238000002716 delivery method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 210000001508 eye Anatomy 0.000 description 1
- 230000010030 glucose lowering effect Effects 0.000 description 1
- 230000002641 glycemic effect Effects 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000004026 insulin derivative Substances 0.000 description 1
- 229960002869 insulin glargine Drugs 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 229940093561 novox Drugs 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 230000037317 transdermal delivery Effects 0.000 description 1
Classifications
-
- 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
- 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
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
-
- 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
Abstract
The invention discloses insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and a preparation method thereof. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are prepared through a grafting reaction and are soaked in insulin solution to prepare insulin-loaded double-modified epsilon-polylysine particles. The preparation method disclosed by the invention is simple; the double modified epsilon-polylysine particles loaded with insulin have low insulin burst release performance, can automatically adjust the release dosage of insulin according to the real-time blood glucose level, can effectively relieve hyperglycemia symptoms, and simultaneously avoid the problem of hypoglycemia caused by excessive insulin; insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles can deliver insulin by subcutaneous injection and transdermal microneedle delivery.
Description
Technical Field
The invention belongs to the technical field of high polymer materials, and particularly relates to insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles and a preparation method thereof.
Background
Diabetes is a chronic disease characterized by hyperglycemia, and there are 5.37 million diabetics worldwide (international diabetes consortium 2021 data). The cause of diabetes is the impaired or complete loss of insulin secretion function, which may also lead to impaired heart, nerve, eye, kidney, etc. function with progression of the disease, thus threatening life safety. Exogenous insulin administration can significantly alleviate hyperglycemia symptoms of diabetics, but the problem of inaccurate dosage during treatment is a main cause of poor glycemic control of diabetics, wherein hypoglycemia caused by overdose is particularly fatal. Therefore, exogenous insulin preparations that can alleviate the symptoms of hyperglycemia and avoid the problem of hypoglycemia are a hotspot in the field of diabetes treatment. Currently, long acting insulins are used clinically, such as commercially available insulin detete, insulin glargine, insulin deglutition, etc. The principle of the treatment is to avoid the rapid lowering of blood glucose level to hypoglycemia during the blood glucose lowering process by lowering the release rate of insulin. However, the problem of hypoglycemia still occurs when long acting insulin is administered in high doses.
Recently, strategies utilizing glucose-sensitive phenylboronic acid-based polymer particles to support insulin have attracted considerable attention in the field of diabetes treatment. This class of polymer particles has an amphiphilic structure in which hydrophobic phenylboronic acid groups can be combined with glucose to form hydrophilic glucose/phenylboronic acid complex groups, thereby inducing swelling or dissolution of the phenylboronic acid based polymer particles and release of the supported insulin. Based on the principle, when the blood sugar level is higher, the glucose-sensitive phenylboronic acid-based polymer particles loaded with insulin can release insulin more quickly, so that the hyperglycemia symptom can be relieved quickly; in the case where the blood glucose level is normal or low, the insulin-loaded glucose-sensitive phenylboronic acid-based polymer particles have the property of inhibiting insulin release, and thus occurrence of hypoglycemia can be avoided. At present, the phenylboronic acid based polymer particles have the problems of complex synthesis process and high mass production cost. For example, the synthesis of polyethylene glycol modified phenylboronic acid based polymer particles as reported by Li et al [1] requires at least 5 steps, while the synthesis of starch modified phenylboronic acid based polymer particles as reported by Wen et al [2] requires 3 steps.
Citation document:
[1]C.Li,F.Huang,Y.Liu,et al.Nitrilotriacetic Acid-Functionalized Glucose-Responsive Complex Micelles for the Efficient Encapsulation and Self-Regulated Release of Insulin Langmuir 2018,34,12116.
[2]N.Wen,S.Lv,C.Gao,et al.Glucose-responsive zwitterionic dialdehyde starch-based micelles with potential anti-phagocytic behavior for insulin delivery Chem.Eng.J.2018,335,52.
disclosure of Invention
In order to solve the problems in the background technology, the invention provides an insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particle and a preparation method thereof. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are prepared by adopting a grafting reaction, and the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin are prepared by soaking the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles in insulin solution.
The technical scheme adopted by the invention is as follows:
1. insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles
The phenylboronic acid/fatty acid double-modified epsilon-polylysine is obtained by grafting fatty acid and 4-carboxyl-3-fluorobenzeneboronic acid on epsilon-polylysine by utilizing a grafting reaction, and the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles can be used for delivering insulin through a subcutaneous injection method after insulin is loaded, or can be used for delivering insulin through a transdermal administration method after the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin are filled in a polymer microneedle patch.
The epsilon-polylysine has the following structural formula:
the phenylboronic acid/fatty acid double modified epsilon-polylysine has the following structural formula:
where n=4, 6, 8, 10, 12, 14, 16 or 18.
2. The preparation method of the insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles comprises the following steps:
1) And (3) adding the fatty acid, the 4-carboxyl-3-fluorobenzeneboronic acid and the auxiliary materials into a solvent, and stirring to obtain a mixture A.
2) Epsilon-polylysine is put into a solvent and stirred to obtain a mixture B.
3) The mixture A and the mixture B are mixed and stirred to obtain a mixture C.
4) The mixture C is filled into a dialysis bag, and the dialysis bag filled with the mixture C is placed into water for dialysis, and the dialysis water is changed into fresh water at intervals. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
5) And soaking phenylboronic acid/fatty acid double-modified epsilon-polylysine in an insulin solution, centrifuging, removing supernatant, collecting solids and freeze-drying to obtain insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles.
The auxiliary materials in the step 1) are one or more of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide.
The fatty acid in the step 1) is one or more of n-butyric acid, n-caproic acid, n-caprylic acid, n-capric acid, lauric acid, myristic acid, palmitic acid and stearic acid, preferably n-capric acid.
The stirring speed in the step 1) is 200-600rpm, the time is 1-240min, and the temperature is normal temperature.
The solvent in the step 1) and the step 2) is one or more of water, dimethyl sulfoxide, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dichloromethane and trichloromethane.
In the step 1), 1 part of 4-carboxyl-3-fluorobenzeneboronic acid, 0.02-5 parts of fatty acid, 0.2-10 parts of auxiliary materials and 10-300 parts of solvent are calculated according to parts by weight.
In the step 2), the epsilon-polylysine is 1 part by weight and the solvent is 1-100 parts by weight.
The stirring speed in the step 2) is 200-600rpm, the time is 1-240min, and the temperature is normal temperature.
The carboxyl modified phenylboronic acid in the step 1) and the epsilon-polylysine in the step 2) are 1 part by mass of 4-carboxyl-3-fluorobenzeneboronic acid and 0.1-10 parts by mass of epsilon-polylysine.
The stirring speed in the step 3) is 200-600rpm, the time is 5-3000min, and the temperature is normal temperature.
The molecular weight cut-off of the dialysis bag in the step 4) is 500-3500Da.
The time interval of each water change in the step 4) is 2-24 hours, the number of water changes is 6-10, and the volume of the water for dialysis is 3-5 liters.
In the step 5), 1 part of phenylboronic acid/fatty acid double modified epsilon-polylysine particles and 0.01-1 part of insulin are calculated according to parts by weight.
The solvent of the insulin solution in the step 5) is water, the insulin is one or more of pig insulin and human insulin, and the mass concentration of the insulin is 0.01-10%.
The soaking time in the step 5) is 10-240min, the rotating speed is 100-10000rpm, and the centrifuging time is 1-60min.
The steps 1) to 3) can be combined into one step, and the fatty acid, the 4-carboxyl-3-fluorobenzeneboronic acid, the auxiliary material, the epsilon-polylysine and the solvent are mixed and stirred to obtain a mixture C. The stirring speed is 200-600rpm, the stirring time is 5-3000min, and the stirring temperature is normal temperature. 1 part of 4-carboxyl-3-fluorobenzeneboronic acid, 0.02-5 parts of fatty acid, 0.2-10 parts of auxiliary material, 0.1-10 parts of epsilon-polylysine and 10-300 parts of solvent.
In the insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles, epsilon-polylysine is modified by hydrophobic fatty acid and phenylboronic acid, and self-assembly is carried out to form the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles, and insulin can be loaded in the particles through hydrophobic action. Since the hydrophobic phenylboronic acid groups can bind to glucose and become hydrophilic glucose/phenylboronic acid complex groups, the particles undergo swelling behavior under the induction of glucose, releasing the internally entrapped insulin. When the blood sugar level is higher, the phenylboronic acid/fatty acid double modified polymer particles loaded with insulin can release insulin faster, and the hyperglycemia symptom can be relieved quickly; at normal or low blood glucose levels, the insulin-loaded phenylboronic acid/fatty acid double modified polymer particles have the property of inhibiting insulin release, and thus the occurrence of hypoglycemia can be avoided. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin can be directly injected subcutaneously, or the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin can be filled in a polymer microneedle patch to prepare the polymer microneedle patch filled with the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin, and the delivery is carried out by a transdermal delivery method, so that diabetics can freely select the delivery mode according to own requirements.
The invention has the beneficial effects that:
1. the synthesis process of phenylboronic acid/fatty acid double-modified epsilon-polylysine is simple, the synthesis reaction only needs 1 step, and the large-scale production cost is low.
2. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles can be completely hydrolyzed into fatty acid, 4-carboxyl-3-fluorobenzeneboronic acid and lysine small molecules, and are beneficial to the excretion of the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles in vivo.
3. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin have stimulus-response performance on glucose concentration, can automatically adjust the release rate and release dosage of the loaded drug according to the change of the glucose concentration, have low insulin burst release performance, can effectively relieve hyperglycemia symptoms, and simultaneously avoid the problem of hypoglycemia caused by excessive insulin.
4. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin have proper hydrophobic stability, and can inhibit the insulin burst release behavior at normal blood sugar level or lower blood sugar level, thereby avoiding the generation of hypoglycemia.
Drawings
FIG. 1 is a nuclear magnetic resonance spectrum of phenylboronic acid/fatty acid double modified epsilon-polylysine prepared in example 4.
FIG. 2 is an insulin accumulation release profile of insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles prepared in example 8.
FIG. 3 is the hypoglycemic duration of insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of example 8 and corresponding comparative examples.
Detailed Description
The invention will be described in further detail with reference to the accompanying drawings and specific examples.
Example 1:
1.2856g of 4-carboxy-3-fluorobenzeneboronic acid, 0.2649g of n-butyric acid, 1.9120g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.1563g N-hydroxysuccinimide are put into 90g of dimethyl sulfoxide and 10g of water and stirred to obtain a mixture A, the stirring speed is 400rpm, the time is 90min, and the temperature is normal temperature. 1.0004g of epsilon-polylysine is put into 10g of water and stirred at 400rpm for 10min at normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at normal temperature to obtain a mixture C. The mixture C is put into a dialysis bag, the molecular weight cut-off of the dialysis bag is 1000Da, the dialysis bag filled with the mixture C is placed into water for dialysis, the dialysis water is changed into fresh water at intervals of 6 hours each time, the water changing times are 9 times, and the volume of the dialysis water is 3 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
Example 2:
1.2705g of 4-carboxy-3-fluorobenzeneboronic acid, 0.2923g of n-hexanoic acid, 1.7997g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.0925g N-hydroxysuccinimide are put into 50g of dimethyl sulfoxide, 25g of N, N-dimethylformamide and 25g of N, N-dimethylacetamide and stirred to obtain a mixture A, the stirring speed is 400rpm, the time is 90min, and the temperature is normal temperature. 1.0202g of epsilon-polylysine is put into 10g of water and stirred at 400rpm for 10min at normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at normal temperature to obtain a mixture C. The mixture C is put into a dialysis bag, the molecular weight cut-off of the dialysis bag is 1000Da, the dialysis bag filled with the mixture C is placed into water for dialysis, the dialysis water is changed into fresh water at intervals of 6 hours each time, the water changing times are 9 times, and the volume of the dialysis water is 4 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
Example 3:
1.3005g of 4-carboxy-3-fluorobenzeneboronic acid, 0.3626g of n-octanoic acid, 1.9001g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.0005g N-hydroxysuccinimide are put into 70g of dimethyl sulfoxide and 30g of tetrahydrofuran and stirred, so that a mixture A is obtained, the stirring speed is 400rpm, the time is 90min and the temperature is normal temperature. 1.0006g of epsilon-polylysine was put into 10g of water and stirred at 400rpm for 10min at room temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at normal temperature to obtain a mixture C. The mixture C is put into a dialysis bag, the molecular weight cut-off of the dialysis bag is 1000Da, the dialysis bag filled with the mixture C is placed into water for dialysis, the dialysis water is changed into fresh water at intervals of 6 hours each time, the water changing times are 9 times, and the volume of the dialysis water is 3 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
Example 4:
1.2808g of 4-carboxy-3-fluorobenzeneboronic acid, 0.3440g of n-decanoic acid, 1.7253g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.0350g N-hydroxysuccinimide are put into 90g of dimethyl sulfoxide and 10g of chloroform and stirred to obtain a mixture A, the stirring speed is 400rpm, the time is 90min, and the temperature is normal temperature. 1.2006g of epsilon-polylysine is put into 10g of water and stirred at 400rpm for 10min at normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 1000min at normal temperature to obtain a mixture C. The mixture C is put into a dialysis bag, the molecular weight cut-off of the dialysis bag is 1000Da, the dialysis bag filled with the mixture C is placed into water for dialysis, the dialysis water is changed into fresh water at intervals of 6 hours each time, the water changing times are 9 times, and the volume of the dialysis water is 3 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
The results of this example are shown in FIG. 1. FIG. 1 is a nuclear magnetic resonance spectrum of phenylboronic acid/fatty acid double modified epsilon-polylysine. The grafting rate of 4-carboxyl-3-fluorobenzeneboronic acid is calculated to be 47%, and the grafting rate of n-capric acid is calculated to be 10%.
Example 5:
1.2884g of 4-carboxy-3-fluorobenzeneboronic acid, 0.6044g of lauric acid, 1.9172g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.1565g N-hydroxysuccinimide were put into 90g of dimethyl sulfoxide and 10g of methylene chloride and stirred to obtain a mixture A at a stirring speed of 400rpm for 90min at room temperature. 1.0009g of epsilon-polylysine was put into 10g of water and stirred at 300rpm for 200min at room temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at normal temperature to obtain a mixture C. The mixture C is put into a dialysis bag, the molecular weight cut-off of the dialysis bag is 1000Da, the dialysis bag filled with the mixture C is placed into water for dialysis, the dialysis water is changed into fresh water at intervals of 6 hours each time, the water changing times are 9 times, and the volume of the dialysis water is 3 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
Example 6:
1.2880g of 4-carboxy-3-fluorobenzeneboronic acid, 0.4050g of myristic acid, 0.4521g of hexadecanoic acid and 5.0129g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride are put into 75g of dimethyl sulfoxide and 25g of chloroform and stirred to obtain a mixture A, the stirring speed is 500rpm, the time is 60min, and the temperature is normal temperature. 1.0009g of epsilon-polylysine was put into 10g of water and 2g of dimethyl sulfoxide and stirred at 400rpm for 10min at room temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at normal temperature to obtain a mixture C. The mixture C is put into a dialysis bag, the molecular weight cut-off of the dialysis bag is 500Da, the dialysis bag filled with the mixture C is placed into water for dialysis, the dialysis water is changed into fresh water at intervals of 8 hours each time, the water changing times are 8 times, and the volume of the dialysis water is 3 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
Example 7:
1.2883g of 4-carboxy-3-fluorobenzeneboronic acid, 1.1317g of octadecanoic acid, 2.1121g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 1.2622g N-hydroxysuccinimide are put into 60g of dimethyl sulfoxide and 40g of methylene chloride and stirred to obtain a mixture A, the stirring speed is 500rpm, the time is 60min, and the temperature is normal temperature. 1.0012g of epsilon-polylysine is put into 10g of water and stirred at 400rpm for 10min at normal temperature to obtain a mixture B. And mixing and stirring the mixture A and the mixture B at the stirring speed of 500rpm for 2000min at normal temperature to obtain a mixture C. Putting the mixture C into a dialysis bag, putting the dialysis bag with the molecular weight cut-off of 1000Da into water for dialysis, changing the dialysis water into fresh water at intervals of 8 hours each time, changing the water for 8 times, and changing the volume of the dialysis water to 3 liters. And after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double modified epsilon-polylysine.
Example 8:
10.0mg of phenylboronic acid/fatty acid double modified epsilon-polylysine obtained in example 4 was immersed in 1g of 0.2% aqueous solution of porcine insulin at normal temperature for 60min with stirring speed of 250rpm. Then, the mixture was centrifuged at 3000rpm for 20min. The supernatant was removed, and the solid was collected and lyophilized to obtain insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles with an insulin loading of 0.151 (mg insulin)/(mg insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles).
As shown in FIG. 2, the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles have an insulin accumulation release profile, which simulates high blood glucose levels with a glucose concentration of 400mg/dL, normal blood glucose levels with a glucose concentration of 100mg/dL, and a glucose concentration of 0mg/dL as a reference. The higher the glucose concentration, the more insulin is released; at normal blood glucose levels, insulin release rates account for only less than 30% of total insulin loading. Insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of 0.06mg are delivered subcutaneously into diabetic SD rats weighing 200g, lowering high blood glucose levels of 500mg/dL to normal blood glucose levels of 50-200mg/dL within 2 hours, after which the normal blood glucose levels can be maintained for more than 12 hours.
Example 9:
12.1mg of phenylboronic acid/fatty acid double modified epsilon-polylysine obtained in example 3 was immersed in 1g of an aqueous solution containing 0.1% of human insulin and 0.2% of porcine insulin at normal temperature for 90min with a stirring rate of 300rpm. Then, the mixture was centrifuged at 3500rpm for a centrifugation time of 15 minutes. Removing the supernatant, collecting the solid and freeze-drying to obtain the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles.
Comparative example 1:
1.0780g of 4-carboxy-3-fluorobenzeneboronic acid, 1.1245g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 0.6739g N-hydroxysuccinimide are put into 100mL of dimethyl sulfoxide and stirred to obtain a mixture A, the stirring speed is 400rpm, the stirring time is 90min, and the stirring temperature is normal temperature. 1.0001g of epsilon-polylysine was put into 10mL of water and stirred at 400rpm for 10min at room temperature to give a mixture B. The mixture A and the mixture B were mixed and stirred at a stirring speed of 500rpm for 2000 minutes at a stirring temperature of 30℃to give a mixture C. Putting the mixture C into a dialysis bag, putting the dialysis bag with the molecular weight cut-off of 1000Da into water for dialysis, and changing the dialysis water into fresh water at intervals of 6 hours each time, wherein the number of water changing times is 9. After the dialysis is finished, the mixture in the dialysis bag is freeze-dried, and the phenylboronic acid modified epsilon-polylysine is obtained, and the grafting rate of phenylboronic acid groups is 65%. 10.0mg of the phenylboronic acid modified epsilon-polylysine is soaked in 1mL of 0.128% human insulin aqueous solution for 60min at normal temperature with stirring speed of 250rpm. Then, the mixture was centrifuged at 3000rpm for 20min. The supernatant was removed, and the solid was collected and lyophilized to obtain insulin-loaded phenylboronic acid modified epsilon-polylysine particles with an insulin loading of 0.102 (mg insulin)/(mg insulin-loaded phenylboronic acid based epsilon-polylysine particles).
The insulin-loaded phenylboronic acid modified epsilon-polylysine particles of comparative example 1 had an insulin accumulation release rate balance of (27.8.+ -. 1.3)% (0 mg/dL) and (36.6.+ -. 1.5)% (100 mg/dL); the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of example 8 had insulin cumulative release rate balance values of (16.0.+ -. 3.7)% (0 mg/dL) and (25.2.+ -. 3.2)% (100 mg/dL). It can be seen that under normal blood glucose (100 mg/dL) or lower blood glucose conditions, insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles have weaker insulin release behavior than insulin-loaded phenylboronic acid modified epsilon-polylysine particles, and thus can better avoid the occurrence of hypoglycemia.
FIG. 3 is the hypoglycemic duration of insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of example 8 and corresponding comparative examples. Wherein INS is pig insulin, the dosage is 0.06mg, and the titer is 1.65 units; the Det-INS is commercially available Novox and insulin deltoid for relieving hypoglycemia symptoms in insulin treatment, the dosage is 0.165mg of insulin deltoid, and the potency is 1.65 units; PBA-PL is an insulin-loaded phenylboronic acid modified epsilon-polylysine granule of comparative example 1, which contains 0.05mg of porcine insulin and has a potency of 1.38 units; PBA/DA-PL was an insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine granule of example 8 containing 0.06mg of porcine insulin with a potency of 1.65 units. After four are delivered into diabetic SD rats weighing 200g by subcutaneous injection, the duration of hypoglycemia (< 70 mg/dL) of the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles is lower than that of both commercial porcine insulin and commercial norand prandial insulin for alleviating hypoglycemia, and lower than that of the insulin-loaded phenylboronic acid modified epsilon-polylysine particles, demonstrating that the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles can effectively avoid the symptoms of hypoglycemia in insulin treatment.
Claims (4)
1. The insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are characterized in that the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are obtained by grafting 4-carboxyl-3-fluorobenzeneboronic acid and fatty acid on epsilon-polylysine through a grafting reaction; insulin delivery is carried out by subcutaneous injection after the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are loaded with insulin, or the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin are loaded in a polymer microneedle patch and insulin delivery is carried out by transdermal administration;
the preparation method of the insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles comprises the following steps:
1) Adding fatty acid, 4-carboxyl-3-fluorobenzeneboronic acid and auxiliary materials into a solvent, and stirring at normal temperature to obtain a mixture A;
2) Adding epsilon-polylysine into a solvent, and stirring at normal temperature to obtain a mixture B;
3) Mixing the mixture A and the mixture B, and stirring at normal temperature to obtain a mixture C;
4) Putting the mixture C into a dialysis bag with the molecular weight cut-off of 500-3500Da, putting the dialysis bag into water for dialysis, changing dialysis water once every 2-24 hours, changing the water for 6-10 times, changing the volume of dialysis water to 3-5 liters, and freeze-drying the mixture in the dialysis bag after the dialysis is finished to obtain phenylboronic acid/fatty acid double modified epsilon-polylysine;
5) Soaking phenylboronic acid/fatty acid double-modified epsilon-polylysine in insulin solution for 10-240min at normal temperature, centrifuging for 1-60min at a rotating speed of 100-10000rpm, removing supernatant, collecting solid and freeze-drying to obtain insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles;
in the step 1), the auxiliary materials are 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide; the fatty acid is n-capric acid;
the solvent in the step 1) is dimethyl sulfoxide and chloroform; the solvent in the step 2) is water;
in the step 1), 1 part of 4-carboxyl-3-fluorobenzeneboronic acid, 0.02-5 parts of fatty acid, 0.2-10 parts of auxiliary materials and 10-300 parts of solvent are calculated according to parts by weight;
in the step 2), the epsilon-polylysine is 1 part by weight and the solvent is 1-100 parts by weight;
the 4-carboxyl-3-fluorobenzeneboronic acid in the step 1) and the epsilon-polylysine in the step 2) are 1 part by mass, and the epsilon-polylysine is 0.1-10 parts by mass.
2. The insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of claim 1 wherein: the stirring speed of the step 1) and the step 2) is 200-600rpm, and the stirring time is 1-240min; the stirring speed of the step 3) is 200-600rpm, and the stirring time is 5-3000min.
3. The insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of claim 1 wherein: in the step 5), 1 part of phenylboronic acid/fatty acid double modified epsilon-polylysine particles and 0.01-1 part of insulin are calculated according to parts by weight.
4. The insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of claim 1 wherein: the solvent of the insulin solution in the step 5) is water, the insulin is one or two of pig insulin and human insulin, and the mass concentration of the insulin is 0.01-10%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9828445B1 (en) * | 2016-12-13 | 2017-11-28 | King Saud University | Synthesis of modified chitosan particles for oral insulin delivery |
| CN111763317A (en) * | 2019-03-13 | 2020-10-13 | 华东师范大学 | Polymer material modified by phenylboronic acid and application thereof in intracellular delivery of protein and polypeptide |
| CN113209028A (en) * | 2021-03-31 | 2021-08-06 | 浙江大学 | Insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles and preparation method thereof |
| CN113230388A (en) * | 2021-03-31 | 2021-08-10 | 浙江大学 | Microneedle patch containing insulin-loaded phenylboronic acid group epsilon-polylysine and preparation method thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9828445B1 (en) * | 2016-12-13 | 2017-11-28 | King Saud University | Synthesis of modified chitosan particles for oral insulin delivery |
| CN111763317A (en) * | 2019-03-13 | 2020-10-13 | 华东师范大学 | Polymer material modified by phenylboronic acid and application thereof in intracellular delivery of protein and polypeptide |
| CN113209028A (en) * | 2021-03-31 | 2021-08-06 | 浙江大学 | Insulin-loaded glucose-sensitive phenylboronic acid group epsilon-polylysine particles and preparation method thereof |
| CN113230388A (en) * | 2021-03-31 | 2021-08-10 | 浙江大学 | Microneedle patch containing insulin-loaded phenylboronic acid group epsilon-polylysine and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Glucose-responsive insulin activity by covalent modification with aliphatic phenylboronic acid conjugates;Chou DH等;Proc Natl Acad;第112卷(第8期);2401-2406 * |
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