CN114632145A - 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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- CN114632145A CN114632145A CN202210218932.8A CN202210218932A CN114632145A CN 114632145 A CN114632145 A CN 114632145A CN 202210218932 A CN202210218932 A CN 202210218932A CN 114632145 A CN114632145 A CN 114632145A
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- insulin
- polylysine
- acid
- fatty acid
- epsilon
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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 165
- 102000004877 Insulin Human genes 0.000 title claims abstract description 102
- 108090001061 Insulin Proteins 0.000 title claims abstract description 102
- 229940125396 insulin Drugs 0.000 title claims abstract description 102
- 108010039918 Polylysine Proteins 0.000 title claims abstract description 101
- 235000014113 dietary fatty acids Nutrition 0.000 title claims abstract description 84
- 229930195729 fatty acid Natural products 0.000 title claims abstract description 84
- 239000000194 fatty acid Substances 0.000 title claims abstract description 84
- 150000004665 fatty acids Chemical class 0.000 title claims abstract description 84
- 239000002245 particle Substances 0.000 title claims abstract description 72
- 238000002360 preparation method Methods 0.000 title 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 83
- 238000000502 dialysis Methods 0.000 claims description 77
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 53
- 238000003756 stirring Methods 0.000 claims description 50
- 238000000034 method Methods 0.000 claims description 26
- 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 21
- 229920000642 polymer Polymers 0.000 claims description 15
- 239000002904 solvent Substances 0.000 claims description 14
- 238000004108 freeze drying Methods 0.000 claims description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 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
- 238000002156 mixing Methods 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 8
- 108010005991 Pork Regular Insulin Proteins 0.000 claims description 7
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 6
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 claims description 6
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims description 6
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 claims description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 5
- 101000976075 Homo sapiens Insulin Proteins 0.000 claims description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 229960001701 chloroform Drugs 0.000 claims description 4
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 claims description 4
- 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
- 239000005639 Lauric acid Substances 0.000 claims description 3
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 claims description 3
- 235000021314 Palmitic acid Nutrition 0.000 claims description 3
- 235000021355 Stearic acid Nutrition 0.000 claims description 3
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 claims description 3
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 3
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 3
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 claims description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 3
- 239000008117 stearic acid Substances 0.000 claims description 2
- 238000013271 transdermal drug delivery Methods 0.000 claims 1
- 239000008280 blood Substances 0.000 abstract description 16
- 210000004369 blood Anatomy 0.000 abstract description 16
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 15
- 208000013016 Hypoglycemia Diseases 0.000 abstract description 13
- 208000024891 symptom Diseases 0.000 abstract description 8
- 201000001421 hyperglycemia Diseases 0.000 abstract description 7
- 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 description 18
- 239000008103 glucose Substances 0.000 description 18
- 238000011049 filling Methods 0.000 description 9
- 239000013505 freshwater Substances 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 206010012601 diabetes mellitus Diseases 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 5
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- 238000003786 synthesis reaction Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000001186 cumulative effect Effects 0.000 description 4
- 230000003914 insulin secretion Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 3
- -1 phenylboronic acid compound Chemical group 0.000 description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 108010092217 Long-Acting Insulin Proteins 0.000 description 2
- 102000016261 Long-Acting Insulin Human genes 0.000 description 2
- 229940100066 Long-acting insulin Drugs 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
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- 229920002085 Dialdehyde starch Polymers 0.000 description 1
- 102100023915 Insulin Human genes 0.000 description 1
- 108010089308 Insulin Detemir Proteins 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
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 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
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003095 anti-phagocytic effect Effects 0.000 description 1
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- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
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- 210000001508 eye Anatomy 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 230000003345 hyperglycaemic effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 229960003948 insulin detemir Drugs 0.000 description 1
- 229960002869 insulin glargine Drugs 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- UGOZVNFCFYTPAZ-IOXYNQHNSA-N levemir Chemical compound CCCCCCCCCCCCCC(=O)NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)CNC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H]1NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=2C=CC(O)=CC=2)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=2N=CNC=2)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=2N=CNC=2)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=2C=CC=CC=2)C(C)C)CSSC[C@@H]2NC(=O)[C@@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)C(C)C)CSSC[C@H](NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CO)NC(=O)[C@H]([C@@H](C)O)NC2=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)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=2C=CC(O)=CC=2)C(=O)N[C@@H](CSSC1)C(=O)N[C@@H](CC(N)=O)C(O)=O)CC1=CC=C(O)C=C1 UGOZVNFCFYTPAZ-IOXYNQHNSA-N 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
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Images
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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Diabetes (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Endocrinology (AREA)
- Dermatology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Emergency Medicine (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
- Immunology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses 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 through a grafting reaction and are soaked in an insulin solution to prepare the 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 the insulin according to the real-time blood sugar level, can effectively relieve hyperglycemia symptoms, and simultaneously avoid hypoglycemia caused by excessive insulin; the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin can deliver insulin by adopting subcutaneous injection and microneedle transdermal administration.
Description
Technical Field
The invention belongs to the technical field of high polymer materials, and particularly relates to an insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particle and a preparation method thereof.
Background
Diabetes is a chronic disease with hyperglycemia as a main characteristic, and 5.37 hundred million diabetics (2021 year data of international diabetes union) exist in the world. The predisposing factor of diabetes is the damage or complete loss of insulin secretion function, which may also lead to the damage of heart, nerve, eye, kidney and other functions along with the progress of the disease, further threatening the life safety. Exogenous insulin can significantly relieve hyperglycemia symptoms of diabetics, but inaccurate dosage in the treatment process is a main reason for poor blood sugar control of diabetics, and low blood sugar caused by overhigh dosage is fatal. Therefore, exogenous insulin preparations that can both alleviate the symptoms of hyperglycemia and avoid the problem of hypoglycemia are a hot spot in the field of diabetes treatment. Currently, long-acting insulin is clinically used for treatment, such as commercialized insulin detemir, insulin glargine, insulin deglutition and the like. The treatment principle is to reduce the release rate of insulin to avoid the rapid reduction of blood sugar level to hypoglycemia during the process of blood sugar reduction. However, long acting insulin is still associated with hypoglycemia at high doses.
Recently, strategies for loading insulin with glycopyrronics based polymer particles have attracted considerable attention in the field of diabetes therapy. The polymer particles have an amphiphilic structure, wherein a hydrophobic phenylboronic acid group can be combined with glucose to form a hydrophilic glucose/phenylboronic acid compound group, so that swelling or dissolution of the phenylboronic acid group polymer particles is induced, and loaded insulin is released. 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, and the hyperglycemia symptom can be relieved quickly; under the condition that the blood sugar level is normal or low, the insulin-loaded glycophenylboronic acid-based polymer particles have the property of inhibiting the release of insulin, so that the occurrence of hypoglycemia can be avoided. At present, the phenylboronic acid-based polymer particles have the problems of complex synthesis process and high large-scale production cost. For example, Li et al [1] reported synthesis of polyethylene glycol-modified phenylboronic acid-based polymer particles required at least 5 steps, while Wen et al [2] reported synthesis of starch-modified phenylboronic acid-based polymer particles required 3 steps.
Citation documents:
[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 art, the invention provides an insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particle and a preparation method thereof. The preparation method comprises the steps of preparing phenylboronic acid/fatty acid double-modified epsilon-polylysine particles by adopting a grafting reaction, and soaking the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles in an insulin solution to prepare the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin.
The technical scheme adopted by the invention is as follows:
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 through 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 being loaded with insulin, and can also be used for delivering insulin through a transdermal administration method after being filled in a polymer microneedle patch.
The epsilon-polylysine has the following structural formula:
the structural formula of the phenylboronic acid/fatty acid double-modified epsilon-polylysine is as follows:
wherein n is 4, 6, 8, 10, 12, 14, 16 or 18.
Secondly, the preparation method of the phenylboronic acid/fatty acid double-modified epsilon-polylysine particle loaded with the insulin comprises the following steps:
1) adding fatty acid, 4-carboxyl-3 fluorobenzeneboronic acid and auxiliary materials into a solvent, and stirring to obtain a mixture A.
2) And (3) adding the epsilon-polylysine into the solvent and stirring to obtain a mixture B.
3) The mixture A and the mixture B are mixed and stirred to obtain a mixture C.
4) And filling the mixture C into a dialysis bag, putting the dialysis bag filled with the mixture C into water for dialysis, and replacing the dialysis water with fresh water at intervals. And after dialysis, freeze-drying the mixture in the dialysis bag to obtain the product phenylboronic acid/fatty acid double-modified epsilon-polylysine.
5) Soaking the phenylboronic acid/fatty acid double-modified epsilon-polylysine in an insulin solution, centrifuging, removing a supernatant, collecting solids, and freeze-drying to obtain the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin.
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, and n-capric acid is preferred.
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 steps 1) and 2) is one or more of water, dimethyl sulfoxide, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dichloromethane and trichloromethane.
In the step 1), by mass, 1 part of 4-carboxyl-3 fluorobenzeneboronic acid, 0.02-5 parts of fatty acid, 0.2-10 parts of auxiliary material and 10-300 parts of solvent are used.
In the step 2), the epsilon-polylysine accounts for 1 part by mass, and the solvent accounts for 1-100 parts by mass.
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 counted by mass parts, wherein the 4-carboxyl-3 fluorobenzeneboronic acid is 1 part, and the epsilon-polylysine is 0.1-10 parts.
The stirring speed in the step 3) is 200-600rpm, the time is 5-3000min, and the temperature is normal temperature.
The cut-off molecular weight of the dialysis bag in the step 4) is 500-.
The time interval of each water change in the step 4) is 2-24h, the water change times are 6-10 times, and the volume of dialysis water is 3-5 liters.
In the step 5), the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are 1 part by mass, and the insulin is 0.01-1 part by mass.
The solvent of the insulin solution in the step 5) is water, the insulin is one or more of porcine 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 centrifugation time is 1-60 min.
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 time is 5-3000min, and the temperature is normal temperature. By mass, 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 phenylboronic acid/fatty acid double-modified epsilon-polylysine particle for loading insulin, epsilon-polylysine is modified by hydrophobic fatty acid and phenylboronic acid and self-assembled to form the phenylboronic acid/fatty acid double-modified epsilon-polylysine particle, and the insulin can be loaded in the particle through a hydrophobic effect. Since the hydrophobic phenylboronic acid group can bind to glucose and change to a hydrophilic glucose/phenylboronic acid complex group, the particles undergo a swelling behavior under the induction of glucose, thereby releasing the insulin entrapped inside. When the blood sugar level is higher, the phenylboronic acid/fatty acid double-modified polymer particles loaded with insulin can release insulin more quickly, and the hyperglycemia symptom can be relieved quickly; under the condition that the blood sugar level is normal or low, the phenylboronic acid/fatty acid double-modified polymer particles loaded with insulin have the property of inhibiting the release of insulin, so that the occurrence of hypoglycemia can be avoided. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with the insulin can be directly injected subcutaneously, and the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with the insulin can also 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 the insulin, and the polymer microneedle patch is delivered by a transdermal administration method, so that a diabetic can freely select an administration mode according to the self requirement.
The invention has the beneficial effects that:
1. the synthesis process of the 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-fluorophenylboronic acid and lysine micromolecules, and the excretion of the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles in vivo is facilitated.
3. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin have stimulation-response performance on glucose concentration, can automatically adjust the release rate and release dosage of loaded drugs according to the change of the glucose concentration, have low insulin burst release performance, can effectively relieve hyperglycemia symptoms, and simultaneously avoid hypoglycemia caused by excessive insulin.
4. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin have proper hydrophobic stability, can inhibit the sudden release behavior of the insulin under normal blood sugar level or lower blood sugar level, and further avoid the generation of hypoglycemia.
Drawings
FIG. 1 is the nuclear magnetic hydrogen spectrum of phenylboronic acid/fatty acid double modified epsilon-polylysine prepared in example 4.
Fig. 2 is an insulin cumulative release profile of insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles prepared in example 8.
Fig. 3 is the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of example 8 and the corresponding comparative ratio of hypoglycemic duration.
Detailed Description
The invention is described in further detail below with reference to the figures and the embodiments.
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 to be stirred, so that a mixture A is obtained, the stirring speed is 400rpm, the time is 90min, and the temperature is normal temperature. 1.0004g of epsilon-polylysine was 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h, wherein the water changing time interval is 9 times, and the volume of the dialysis water is 3 liters. And after dialysis, 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-carboxyl-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 N, N-dimethylformamide and 25g N, N-dimethylacetamide and stirred to obtain a mixture A, wherein the stirring speed is 400rpm, the time is 90min, and the temperature is normal temperature. 1.0202g of epsilon-polylysine was 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h, wherein the water changing time interval is 9 times, and the volume of the dialysis water is 4 liters. And after dialysis, 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 to obtain a mixture A, wherein the stirring speed is 400rpm, the stirring time is 90min, and the temperature is normal temperature. 1.0006g of epsilon-polylysine is put into 10g of water and stirred, the stirring speed is 400rpm, the time is 10min, and the temperature is normal temperature, so that a mixture B is obtained. 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h, wherein the water changing time interval is 9 times, and the volume of the dialysis water is 3 liters. And after dialysis, 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-carboxyl-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 trichloromethane and stirred to obtain a mixture A, wherein the stirring speed is 400rpm, the stirring time is 90min, and the temperature is normal temperature. 1.2006g of epsilon-polylysine was 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h, wherein the water changing time interval is 9 times, and the volume of the dialysis water is 3 liters. And after dialysis, 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 hydrogen spectrum of phenylboronic acid/fatty acid double-modified epsilon-polylysine. The calculated grafting rate of the 4-carboxyl-3-fluorobenzeneboronic acid is 47 percent, and the grafting rate of the n-decanoic acid is 10 percent.
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 g of hydroxysuccinimide are put into 90g of dimethyl sulfoxide and 10g of dichloromethane and stirred to obtain a mixture A, wherein the stirring speed is 400rpm, the stirring time is 90min, and the temperature is normal temperature. 1.0009g of epsilon-polylysine is put into 10g of water and stirred, the stirring speed is 300rpm, the time is 200min, and the temperature is normal temperature, so that a mixture B is obtained. 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h, wherein the water changing time interval is 9 times, and the volume of the dialysis water is 3 liters. And after dialysis, 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 palmitic acid and 5.0129g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride were put into 75g of dimethyl sulfoxide and 25g of chloroform and stirred to obtain a mixture A, the stirring rate was 500rpm, the time was 60min, and the temperature was room temperature. 1.0009g of epsilon-polylysine is put into 10g of water and 2g of dimethyl sulfoxide and stirred at the speed of 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 500Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 8h, wherein the water changing time interval is 8 times, and the volume of the dialysis water is 3 liters. And after dialysis, 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 g of hydroxysuccinimide are put into 60g of dimethyl sulfoxide and 40g of dichloromethane to be stirred, so that a mixture A is obtained, the stirring speed is 500rpm, the stirring time is 60min, and the temperature is normal temperature. 1.0012g of epsilon-polylysine was 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. And (3) filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 8h, wherein the water changing time interval is 8 times, and the volume of the dialysis water is 3 liters. And after dialysis, 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 di-modified epsilon-polylysine obtained in example 4 was immersed in 1g of 0.2% swine insulin aqueous solution at room temperature for 60min, with a stirring rate of 250 rpm. Then, the mixture was centrifuged at 3000rpm for 20 min. The supernatant was removed, the solids were collected and lyophilized to obtain insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles with an insulin load of 0.151(mg insulin)/(mg insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles).
As shown in FIG. 2, the cumulative release profile of insulin from the insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles was modeled using a glucose concentration of 400mg/dL to simulate high blood glucose levels, a glucose concentration of 100mg/dL to simulate normal blood glucose levels, 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, the rate of insulin release is only less than 30% of the total insulin load. 0.06mg of the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles were delivered by subcutaneous injection into diabetic SD rats weighing 200g, and the hyperglycemic level of 500mg/dL was reduced to a normoglycemic level of 50-200mg/dL within 2 hours, after which the normoglycemic level could be maintained for more than 12 hours.
Example 9:
12.1mg of phenylboronic acid/fatty acid di-modified epsilon-polylysine obtained in example 3 was immersed in 1g of an aqueous solution containing 0.1% human insulin and 0.2% porcine insulin at room temperature for 90min at a stirring speed of 300 rpm. Then, the mixture was centrifuged at 3500rpm for 15 min. And 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, wherein the stirring speed is 400rpm, the stirring time is 90min, and the stirring temperature is normal temperature. And (3) putting 1.0001g of epsilon-polylysine into 10mL of water, and stirring at the speed of 400rpm for 10min at the 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 the stirring temperature of 30 ℃ to obtain a mixture C. And filling the mixture C into a dialysis bag, wherein the cut-off molecular weight of the dialysis bag is 1000Da, putting the dialysis bag filled with the mixture C into water for dialysis, and changing the dialysis water into fresh water at intervals of 6h every time, wherein the water changing times are 9 times. And after dialysis, freeze-drying the mixture in the dialysis bag to obtain a product phenylboronic acid modified epsilon-polylysine, wherein 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 the normal temperature and at the stirring speed of 250 rpm. Then, the mixture was centrifuged at 3000rpm for 20 min. The supernatant was removed, the solids were collected and lyophilized to obtain insulin-loaded phenylboronic acid-modified epsilon-polylysine particles with an insulin load of 0.102(mg insulin)/(mg insulin-loaded phenylboronic acid-epsilon-polylysine particles).
The insulin cumulative release rate equilibrium values of the insulin-loaded phenylboronic acid-modified epsilon-polylysine particles of comparative example 1 were (27.8 + -1.3)% (0mg/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 equilibrium values of cumulative insulin release rates of (16.0 + -3.7)% (0mg/dL) and (25.2 + -3.2)% (100 mg/dL). It can be seen that the phenylboronic acid/fatty acid double-modified epsilon-polylysine-loaded particles have a weaker insulin release behavior than the phenylboronic acid-modified epsilon-polylysine-loaded particles under conditions of normal blood glucose (100mg/dL) or lower blood glucose levels, and therefore, the generation of hypoglycemia can be better avoided.
Fig. 3 is the insulin-loaded phenylboronic acid/fatty acid double modified epsilon-polylysine particles of example 8 and the corresponding comparative ratio of hypoglycemic duration. Wherein INS is porcine insulin, the dosage is 0.06mg, and the potency is 1.65 unit; Det-INS is commercially available insulin Normandiprodione and insulin rague, is used for relieving hypoglycemia symptoms in insulin treatment, and has dosage of 0.165mg insulin rague and potency of 1.65 unit; PBA-PL is phenylboronic acid modified epsilon-polylysine particles loaded with insulin of comparative example 1, containing 0.05mg of porcine insulin, and the titer is 1.38 units; PBA/DA-PL was phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin of example 8, containing 0.06mg porcine insulin, and having a potency of 1.65 units. After four of them were delivered subcutaneously into diabetic SD rats weighing 200g, the duration of hypoglycemia (<70mg/dL) of the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles was lower than both commercially available porcine insulin and commercially available nordheim for the relief of hypoglycemia and than insulin-loaded phenylboronic acid-modified epsilon-polylysine particles, demonstrating that the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles could effectively avoid hypoglycemia symptoms in insulin therapy.
Claims (10)
1. The phenylboronic acid/fatty acid double-modified epsilon-polylysine particle loaded with insulin is characterized in that the phenylboronic acid/fatty acid double-modified epsilon-polylysine particle is obtained by grafting 4-carboxyl-3 fluorobenzeneboronic acid and fatty acid on epsilon-polylysine through a grafting reaction; the phenylboronic acid/fatty acid double-modified group epsilon-polylysine particles are loaded with insulin and then delivered by a subcutaneous injection method, or the phenylboronic acid/fatty acid double-modified group epsilon-polylysine particles loaded with the insulin are filled in a polymer microneedle patch and delivered by a transdermal drug delivery method.
2. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 1, comprising the steps of:
1) putting 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 cutoff of 500-3500Da, placing the bag in water for dialysis, changing dialysis water once every 2-24h, wherein the water changing frequency is 6-10 times, and the volume of the dialysis water is 3-5 liters, and after the dialysis is finished, freeze-drying the mixture in the dialysis bag to obtain phenylboronic acid/fatty acid double-modified epsilon-polylysine;
5) soaking the phenylboronic acid/fatty acid double-modified epsilon-polylysine in an insulin solution at normal temperature for 10-240min, centrifuging at the rotation speed of 100 plus 10000rpm for 1-60min, removing supernatant, collecting solids, and freeze-drying to obtain the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles loaded with insulin.
3. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: in the step 1), the auxiliary material is one or more of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide; the fatty acid 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.
4. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: the solvent in the steps 1) and 2) is one or more of water, dimethyl sulfoxide, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dichloromethane and trichloromethane.
5. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: the stirring speed of the step 1) and the step 2) is 200-600rpm, and the stirring time is 1-240 min; the stirring speed of the step 3) is 200-600rpm, and the stirring time is 5-3000 min.
6. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: in the step 1), by mass, 1 part of 4-carboxyl-3 fluorobenzeneboronic acid, 0.02-5 parts of fatty acid, 0.2-10 parts of auxiliary material and 10-300 parts of solvent are used.
7. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: in the step 2), the epsilon-polylysine accounts for 1 part by mass, and the solvent accounts for 1-100 parts by mass.
8. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: the 4-carboxyl-3 fluorobenzeneboronic acid in the step 1) and the epsilon-polylysine in the step 2) are counted by mass parts, wherein the 4-carboxyl-3 fluorobenzeneboronic acid is 1 part, and the epsilon-polylysine is 0.1-10 parts.
9. The method for preparing the insulin-loaded phenylboronic acid/fatty acid di-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: in the step 5), the phenylboronic acid/fatty acid double-modified epsilon-polylysine particles are 1 part by weight, and the insulin is 0.01-1 part by weight.
10. The method for preparing the insulin-loaded phenylboronic acid/fatty acid double-modified epsilon-polylysine particles according to claim 2, wherein the method comprises the following steps: the solvent of the insulin solution in the step 5) is water, the insulin is one or more of porcine insulin and human insulin, and the mass concentration of the insulin is 0.01-10%.
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| WO2024067600A1 (en) * | 2022-09-29 | 2024-04-04 | 浙江大学 | Glucose-responsive complex, preparation method therefor, and use thereof |
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| WO2024067600A1 (en) * | 2022-09-29 | 2024-04-04 | 浙江大学 | Glucose-responsive complex, preparation method therefor, and use thereof |
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