CN110694052A - 利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用 - Google Patents
利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用 Download PDFInfo
- Publication number
- CN110694052A CN110694052A CN201911087840.5A CN201911087840A CN110694052A CN 110694052 A CN110694052 A CN 110694052A CN 201911087840 A CN201911087840 A CN 201911087840A CN 110694052 A CN110694052 A CN 110694052A
- Authority
- CN
- China
- Prior art keywords
- liraglutide
- preparation
- peripheral nerve
- application
- nerve
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- YSDQQAXHVYUZIW-QCIJIYAXSA-N Liraglutide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCNC(=O)CC[C@H](NC(=O)CCCCCCCCCCCCCCC)C(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=C(O)C=C1 YSDQQAXHVYUZIW-QCIJIYAXSA-N 0.000 title claims abstract description 31
- 108010019598 Liraglutide Proteins 0.000 title claims abstract description 27
- 229960002701 liraglutide Drugs 0.000 title claims abstract description 27
- 210000000578 peripheral nerve Anatomy 0.000 title claims abstract description 18
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 230000023105 myelination Effects 0.000 title claims abstract description 10
- 230000001737 promoting effect Effects 0.000 title claims abstract description 9
- 239000003814 drug Substances 0.000 claims description 6
- 102000006386 Myelin Proteins Human genes 0.000 claims description 3
- 108010083674 Myelin Proteins Proteins 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 3
- 210000005012 myelin Anatomy 0.000 claims description 3
- 210000003497 sciatic nerve Anatomy 0.000 abstract description 16
- 210000003007 myelin sheath Anatomy 0.000 abstract description 14
- 101500028774 Homo sapiens Glucagon-like peptide 1 Proteins 0.000 abstract description 8
- KISWVXRQTGLFGD-UHFFFAOYSA-N 2-[[2-[[6-amino-2-[[2-[[2-[[5-amino-2-[[2-[[1-[2-[[6-amino-2-[(2,5-diamino-5-oxopentanoyl)amino]hexanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxypropanoyl]amino]-5-oxopentanoyl]amino]-5-(diaminomethylideneamino)p Chemical compound C1CCN(C(=O)C(CCCN=C(N)N)NC(=O)C(CCCCN)NC(=O)C(N)CCC(N)=O)C1C(=O)NC(CO)C(=O)NC(CCC(N)=O)C(=O)NC(CCCN=C(N)N)C(=O)NC(CO)C(=O)NC(CCCCN)C(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=C(O)C=C1 KISWVXRQTGLFGD-UHFFFAOYSA-N 0.000 abstract description 7
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical class C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 abstract description 7
- 230000015572 biosynthetic process Effects 0.000 abstract description 7
- 101150059401 EGR2 gene Proteins 0.000 abstract description 5
- 239000003102 growth factor Substances 0.000 abstract description 3
- 238000005259 measurement Methods 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 9
- 241000700159 Rattus Species 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 6
- 230000014509 gene expression Effects 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 description 4
- 102000047918 Myelin Basic Human genes 0.000 description 4
- 101710107068 Myelin basic protein Proteins 0.000 description 4
- 239000002033 PVDF binder Substances 0.000 description 4
- 210000003050 axon Anatomy 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 210000005036 nerve Anatomy 0.000 description 4
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 108020004463 18S ribosomal RNA Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 108010051748 Early Growth Response Protein 2 Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 208000010886 Peripheral nerve injury Diseases 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- 210000002237 B-cell of pancreatic islet Anatomy 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- GDBQQVLCIARPGH-UHFFFAOYSA-N Leupeptin Natural products CC(C)CC(NC(C)=O)C(=O)NC(CC(C)C)C(=O)NC(C=O)CCCN=C(N)N GDBQQVLCIARPGH-UHFFFAOYSA-N 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 229940100389 Sulfonylurea Drugs 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- ABUBSBSOTTXVPV-UHFFFAOYSA-H [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O Chemical compound [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O ABUBSBSOTTXVPV-UHFFFAOYSA-H 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- HOQPTLCRWVZIQZ-UHFFFAOYSA-H bis[[2-(5-hydroxy-4,7-dioxo-1,3,2$l^{2}-dioxaplumbepan-5-yl)acetyl]oxy]lead Chemical compound [Pb+2].[Pb+2].[Pb+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HOQPTLCRWVZIQZ-UHFFFAOYSA-H 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 239000003822 epoxy resin Substances 0.000 description 1
- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- GDBQQVLCIARPGH-ULQDDVLXSA-N leupeptin Chemical compound CC(C)C[C@H](NC(C)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C=O)CCCN=C(N)N GDBQQVLCIARPGH-ULQDDVLXSA-N 0.000 description 1
- 108010052968 leupeptin Proteins 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 1
- 229960003105 metformin Drugs 0.000 description 1
- 238000003801 milling Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000000944 nerve tissue Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- QNDVLZJODHBUFM-WFXQOWMNSA-N okadaic acid Chemical compound C([C@H](O1)[C@H](C)/C=C/[C@H]2CC[C@@]3(CC[C@H]4O[C@@H](C([C@@H](O)[C@@H]4O3)=C)[C@@H](O)C[C@H](C)[C@@H]3[C@@H](CC[C@@]4(OCCCC4)O3)C)O2)C(C)=C[C@]21O[C@H](C[C@@](C)(O)C(O)=O)CC[C@H]2O QNDVLZJODHBUFM-WFXQOWMNSA-N 0.000 description 1
- VEFJHAYOIAAXEU-UHFFFAOYSA-N okadaic acid Natural products CC(CC(O)C1OC2CCC3(CCC(O3)C=CC(C)C4CC(=CC5(OC(CC(C)(O)C(=O)O)CCC5O)O4)C)OC2C(O)C1C)C6OC7(CCCCO7)CCC6C VEFJHAYOIAAXEU-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229920000647 polyepoxide Polymers 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- GCYXWQUSHADNBF-AAEALURTSA-N preproglucagon 78-108 Chemical class C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 GCYXWQUSHADNBF-AAEALURTSA-N 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 210000004116 schwann cell Anatomy 0.000 description 1
- FQENQNTWSFEDLI-UHFFFAOYSA-J sodium diphosphate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]P([O-])(=O)OP([O-])([O-])=O FQENQNTWSFEDLI-UHFFFAOYSA-J 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000003656 tris buffered saline Substances 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
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
- A61K38/26—Glucagons
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Neurology (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Endocrinology (AREA)
- Biomedical Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Neurosurgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了人胰高糖素样肽‑1类似物利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用。通过对早期生长因子20（krox20）和髓鞘基础蛋白（MBP）的测定，及坐骨神经超微结构的观察，证实了利拉鲁肽可促进外周神经髓鞘的形成。
Description
技术领域
本发明属于医学技术领域，具体涉及人胰高糖素样肽-1类似物利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用。
背景技术
利拉鲁肽(liraglutide，Arg34Lys26-(N-ε-(γ-Glu(N-α-十六酰基))))是一种人胰高糖素样肽-1(GLP-1)类似物，用于成人II型糖尿病患者控制血糖，尤其适用于单用二甲双胍或磺脲类药物治疗效果不佳的患者。该药物能够促进胰腺β细胞葡萄糖浓度依赖性的分泌胰岛素，维持体内血糖稳定，能量平衡。它的刺激会引起体内环磷酸腺苷(cAMP)的增加，很多研究表明，利拉鲁肽的激活可平衡体内细胞血糖稳定，为生理活动提供能量。
外周神经细胞轴突外面包裹着由雪旺细胞组成的膜结构，称之为髓鞘。它的生理作用表现为绝缘，防止神经电冲动从神经元轴突传递至另一神经元轴突。在日常生活中，外周神经损伤是一种常见病征，车祸、地震等各种天灾人祸都有可能造成外周神经损伤，而损伤后髓鞘的形成是神经功能得以恢复的重要保证。大量研究指出，髓鞘形成需要环磷酸腺苷(cAMP)的刺激，而利拉鲁肽的刺激会引起体内环磷酸腺苷(cAMP)的增加，因此，发明人推测利拉鲁肽可能有助于外周神经髓鞘的形成。
发明内容
本发明的目的是提供人胰高糖素样肽-1类似物利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用。
为了实现上述发明目的，本发明采用以下技术方案：
利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用。
利拉鲁肽在制备治疗外周神经髓鞘类疾病药物中的应用。
本发明验证了人胰高糖素样肽-1类似物利拉鲁肽可促进外周神经髓鞘的形成，从而可用于制备促进外周神经髓鞘形成制剂和治疗外周神经髓鞘类疾病药物。
附图说明
图1为实施例1中坐骨神经组织MBP、Krox20基因表达分析。其中，Vehicle为对照组，Liraglutide为利拉鲁肽组。
图2为实施例1中坐骨神经组织MBP、Krox20蛋白表达分析。其中，Vehicle为对照组，Liraglutide为利拉鲁肽组。
图3为实施例1中坐骨神经的电镜图。其中，Vehicle为对照组，Liraglutide为利拉鲁肽组。
图4为大鼠坐骨神经髓鞘厚度分析。****p<0.0001为Student's t test分析。其中，Vehicle为对照组，Liraglutide为利拉鲁肽组。
具体实施方式
下面结合具体实施例，进一步阐述本发明。下述实施例中所使用的实验方法如无特殊说明，均为常规方法。下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到，未详细描述的技术均按照本领域人员熟知的标准方法进行。本申请中提及的试剂等均有商品供应或以别的途径能为公众所得，它们仅作为举例，对本发明不是唯一的。可分别用其它适合的工具或生物材料来替代。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。
实施例1
为研究人胰高糖素样肽-1类似物利拉鲁肽是否可以促进外周神经髓鞘的形成，本发明用其处理刚出生的大鼠(剂量为15μg/kg体重/天，皮下注射)，在处理后的8天收集坐骨神经组织用于后续分析。
1、动物处理
实验材料为刚出生的SD大鼠，向每只大鼠皮下注射利拉鲁肽，剂量为15μg/kg体重/天。对照组大鼠注射等剂量的磷酸缓冲液(PBS)。在处理后8天收集大鼠坐骨神经组织用于后续分析。每组实验动物8只，实验重复三次。
2、坐骨神经组织RNA提取及基因表达检测
使用Trizol试剂(Invitrogen公司)提取坐骨神经组织的RNA，再逆转录合成cDNA第一链(Roche公司)。采用SYBR Green Supermix(Bio-Rad公司)试剂检测各基因表达情况，通过仪器iQ5 Multicolor Real-Time PCR Detection System(Bio-Rad公司产品)实现。数据分析采用
方法来衡量mRNA水平。以18s看家基因来校准mRNA的水平。
所使用的引物序列如下：
18S rRNA正向引物:AGCTCCAATAGCGTATATTAAAG(SEQ ID NO.1)；
18S rRNA负向引物:CGGTCCTATTCCATTATTCCTA(SEQ ID NO.2)；
Krox20正向引物:TGGGTTTAAGTATGGCTGTATA(SEQ ID NO.3)；
Krox20负向引物:AGTTAGTGGTTCTGTGTTAGA(SEQ ID NO.4)；
MBP正向引物:GAGAGTGTGGGTTTAAGATAAC(SEQ ID NO.5)；
MBP负向引物:TCAATATCCAAGAACAGTAGGT(SEQ ID NO.6)。
3、坐骨神经蛋白提取及检测
采用台式匀浆机(Polytron，PT2100)对坐骨神经组织进行磨碎匀浆处理。
动物组织裂解液配方：25mM Tris-HCl，pH 7.4；100mM NaF；50mM Na4P2O7；10mMNa3VO4；10mM EGTA；10mM EDTA；1％NP-40；10μg/mL Leupeptin；10μg/mL Aprotinin；2mMPMSF；20nM Okadaic acid。
样品于4℃旋转裂解1小时后离心(12500rpm)30min，离心后去除上层脂质，上清转移至另一离心管并再次离心。这一过程重复2次以彻底去除蛋白样品中的脂质成分。样品蛋白含量用试剂盒测定(pierce公司产品)，根据所得到的结果将所有样品蛋白浓度调至相同水平，加入上样缓冲液混匀后于100℃煮沸5min，冷却至室温后冷冻保存，用于后续Westernblot分析。
将上述步骤得到的样品用聚丙烯酰胺凝胶电泳(SDS-PAGE)分离，并将胶上蛋白转至PVDF膜上。转膜后的PVDF膜用含5％牛血清白蛋白(BSA)的TBST(Tris-buffered salinesolution/Tween)缓冲液室温封闭1小时。封闭后的PVDF膜与一抗4℃共敷过夜，之后用TBST清洗PVDF膜三次，再与二抗室温孵育1小时，随后用TBST清洗三次。最后用化学发光反应体系(chemiluminescence assay system,Roche)反应，曝光至X光片(Kodak)。蛋白表达水平定量采用Quantity-One(Bio-Rad)分析。所有抗体均购自于Cell Signaling Technology公司。
早期生长因子20(krox20)和髓鞘基础蛋白(MBP)表达升高，是髓鞘形成过程中非常关键的基因。如图1所示，利拉鲁肽可有效激活这些基因的表达。同时，如图2所示，利拉鲁肽处理还可显著促进早期生长因子20(krox20)和髓鞘基础蛋白(MBP)表达的升高。
4、坐骨神经组织结构透射电镜观察
实验组和对照组随机抽取3只动物取坐骨神经作为电镜标本。在动物深度麻醉下，取坐骨神经组织并修切成电镜标本大小(1.2mm×1mm×1mm)，浸入到4％戊二醛溶液进行固定，并用1％的锇酸进行二次固定，醋酸铀染色，梯度乙醇脱水，环氧树脂包埋，半薄切片定位，超薄切片经枸橼酸铅复染，透射电镜选片观察。所得照片在仪器自带图像分析系统上测量有髓神经直径和轴突直径，以轴突直径/有髓神经直径数值(g-ratio)衡量髓鞘厚度，从而推测髓鞘发育情况。
如图3所示，在坐骨神经的超微结构中，利拉鲁肽处理的大鼠坐骨神经髓鞘套厚度较对照组变厚，髓鞘结构明显畸形。图4中的定量数据也表明利拉鲁肽的处理可促进新生大鼠坐骨神经髓鞘的厚度。
以上结果表明，人胰高糖素样肽-1类似物利拉鲁肽可促进外周神经髓鞘的形成。
序列表
<110> 南通大学
<120> 利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用
<130> 20191108
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
agctccaata gcgtatatta aag 23
<210> 2
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
cggtcctatt ccattattcc ta 22
<210> 3
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
tgggtttaag tatggctgta ta 22
<210> 4
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
agttagtggt tctgtgttag a 21
<210> 5
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
gagagtgtgg gtttaagata ac 22
<210> 6
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
tcaatatcca agaacagtag gt 22
Claims (2)
1.利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用。
2.利拉鲁肽在制备治疗外周神经髓鞘类疾病药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911087840.5A CN110694052A (zh) | 2019-11-08 | 2019-11-08 | 利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911087840.5A CN110694052A (zh) | 2019-11-08 | 2019-11-08 | 利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110694052A true CN110694052A (zh) | 2020-01-17 |
Family
ID=69204715
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911087840.5A Pending CN110694052A (zh) | 2019-11-08 | 2019-11-08 | 利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110694052A (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023138320A1 (zh) * | 2022-01-21 | 2023-07-27 | 上海市精神卫生中心（上海市心理咨询培训中心） | 胰岛素样生长因子结合蛋白2在制备治疗神经脱髓鞘性疾病的药物中的应用 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007296A (zh) * | 2012-12-24 | 2013-04-03 | 上海市内分泌代谢病研究所 | 一种利拉鲁肽干预gk大鼠模型及其在制备干预治疗2型糖尿病药物中的应用 |
| CN105920015A (zh) * | 2016-05-09 | 2016-09-07 | 南通大学 | AMPK抑制剂Compound C在制备激活坐骨神经髓鞘形成制剂中的应用 |
-
2019
- 2019-11-08 CN CN201911087840.5A patent/CN110694052A/zh active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007296A (zh) * | 2012-12-24 | 2013-04-03 | 上海市内分泌代谢病研究所 | 一种利拉鲁肽干预gk大鼠模型及其在制备干预治疗2型糖尿病药物中的应用 |
| CN105920015A (zh) * | 2016-05-09 | 2016-09-07 | 南通大学 | AMPK抑制剂Compound C在制备激活坐骨神经髓鞘形成制剂中的应用 |
Non-Patent Citations (4)
| Title |
|---|
| JINGJING MA ET AL.: "GLP‑1R agonists ameliorate peripheral nerve dysfunction and inflammation via p38 MAPK/NF‑κB signaling pathways in streptozotocin‑induced diabetic rats", 《INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE》 * |
| 曾琳等: "大鼠坐骨神经离断后髓鞘碱性蛋白的变化研究", 《中国临床康复》 * |
| 李红志等: "利拉鲁肽对脊髓损伤修复作用的探讨", 《山东大学学报(医学版)》 * |
| 温旭东等: "胰高血糖素样肽-1缓解糖尿病周围神经病变的机制研究进展", 《第二军医大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023138320A1 (zh) * | 2022-01-21 | 2023-07-27 | 上海市精神卫生中心（上海市心理咨询培训中心） | 胰岛素样生长因子结合蛋白2在制备治疗神经脱髓鞘性疾病的药物中的应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Roberts et al. | Mechanisms of mechanical overload-induced skeletal muscle hypertrophy: current understanding and future directions | |
| McKoy et al. | Expression of insulin growth factor‐1 splice variants and structural genes in rabbit skeletal muscle induced by stretch and stimulation | |
| Yariz et al. | Mutations in OTOGL, encoding the inner ear protein otogelin-like, cause moderate sensorineural hearing loss | |
| ITUB20155765A1 (it) | Metodi per diagnosi, prognosi e monitoraggio terapeutico di patologie neurologiche, neurodegenerative e infiammatorie basati su microRNA contenuto in microvescicole microglia | |
| CN103372218B (zh) | 自身免疫性疾病相关的microRNA及其应用 | |
| Kim et al. | Correlation between expression of glucose transporters in granulosa cells and oocyte quality in women with polycystic ovary syndrome | |
| Escobar et al. | Expression of aquaporins early in human pregnancy | |
| Ning et al. | Lycopene‐Loaded Microemulsion Regulates Neurogenesis in Rats with Aβ‐Induced Alzheimer’s Disease Rats Based on the Wnt/β‐catenin Pathway | |
| McKay et al. | Brain-derived neurotrophic factor is associated with human muscle satellite cell differentiation in response to muscle-damaging exercise | |
| Li et al. | Control of growth hormone receptor and insulin‐like growth factor‐I expression by cortisol in ovine fetal skeletal muscle | |
| Makurina et al. | Functional Features of Platelets in Rats Fed a Standard Diet with Low Antioxidant Content During Ontogenesis. | |
| Lai et al. | Expression of cAMP response element-binding protein in major depression before and after antidepressant treatment | |
| CN110694052A (zh) | 利拉鲁肽在制备促进外周神经髓鞘形成制剂中的应用 | |
| CN107937532B (zh) | 胶质瘤诊断标志物hsa_circ_0021827及应用 | |
| CN107557472B (zh) | 胶质瘤诊断标志物circ9:135881633|135883078及应用 | |
| Hou et al. | Activation of the NGF/TrkA signaling pathway attenuates diabetic erectile dysfunction | |
| Heiran et al. | C‐Kit+ progenitors restore rat asthmatic lung function by modulation of T‐bet and GATA‐3 expression | |
| US20220195525A1 (en) | Molecular marker detection and regulating methods in de-servitization state of cells | |
| CN107937538B (zh) | 胶质瘤诊断标志物circ1:201817088|201817285及应用 | |
| Huang et al. | Low-frequency electrical stimulation of the hippocampus plays a role in the treatment of pharmacoresistant epilepsy by blocking the PKA-CREB pathway | |
| CN109820856A (zh) | 化合物Compound C的新应用 | |
| JP2019502381A (ja) | 細胞から指標シグナルを得る方法 | |
| CN107779502A (zh) | Mir‑143在制备用于脑卒中诊断的试剂盒中的应用 | |
| Zhou et al. | Hypoxia-induced regulation of placental REDD1 and mTOR was impaired in a rat model of estrogen-induced cholestasis | |
| CN117327704A (zh) | Npas4在学习记忆功能及突触可塑性损伤中的应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200117 |