CN109125249A - Preparation method and application of tyrosine-derived amygdalin-loaded hydrogel - Google Patents
Preparation method and application of tyrosine-derived amygdalin-loaded hydrogel Download PDFInfo
- Publication number
- CN109125249A CN109125249A CN201810996854.8A CN201810996854A CN109125249A CN 109125249 A CN109125249 A CN 109125249A CN 201810996854 A CN201810996854 A CN 201810996854A CN 109125249 A CN109125249 A CN 109125249A
- Authority
- CN
- China
- Prior art keywords
- amarogentin
- hydrogel
- tyrosine
- tyr
- fmoc
- 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.)
- Granted
Links
- 239000000017 hydrogel Substances 0.000 title claims abstract description 85
- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 title claims abstract description 28
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 title claims abstract description 27
- 229940089837 amygdalin Drugs 0.000 title abstract description 14
- YZLOSXFCSIDECK-UHFFFAOYSA-N amygdalin Natural products OCC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC(C#N)c3ccccc3 YZLOSXFCSIDECK-UHFFFAOYSA-N 0.000 title abstract description 12
- YGHHWSRCTPQFFC-UHFFFAOYSA-N eucalyptosin A Natural products OC1C(O)C(O)C(CO)OC1OC1C(OC(C#N)C=2C=CC=CC=2)OC(CO)C(O)C1O YGHHWSRCTPQFFC-UHFFFAOYSA-N 0.000 title abstract description 12
- XUCIJNAGGSZNQT-JHSLDZJXSA-N (R)-amygdalin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O[C@@H](C#N)C=2C=CC=CC=2)O1 XUCIJNAGGSZNQT-JHSLDZJXSA-N 0.000 title abstract description 11
- 150000003667 tyrosine derivatives Chemical class 0.000 claims abstract description 50
- 208000030886 Traumatic Brain injury Diseases 0.000 claims abstract description 38
- 239000011550 stock solution Substances 0.000 claims abstract description 37
- SWZCTMTWRHEBIN-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(4-hydroxyphenyl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=C(O)C=C1 SWZCTMTWRHEBIN-QFIPXVFZSA-N 0.000 claims abstract description 35
- 206010019196 Head injury Diseases 0.000 claims abstract description 22
- 239000000243 solution Substances 0.000 claims abstract description 18
- 206010061218 Inflammation Diseases 0.000 claims abstract description 13
- 230000004054 inflammatory process Effects 0.000 claims abstract description 13
- 239000000843 powder Substances 0.000 claims abstract description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000007787 solid Substances 0.000 claims abstract description 8
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 7
- 239000012498 ultrapure water Substances 0.000 claims abstract description 7
- DBOVHQOUSDWAPQ-WTONXPSSSA-N amarogentin Chemical compound O([C@H]1[C@H](O[C@H]2[C@@H]([C@H]3C(C(OCC3)=O)=CO2)C=C)O[C@@H]([C@H]([C@@H]1O)O)CO)C(=O)C1=C(O)C=C(O)C=C1C1=CC=CC(O)=C1 DBOVHQOUSDWAPQ-WTONXPSSSA-N 0.000 claims description 87
- DBOVHQOUSDWAPQ-UHFFFAOYSA-N (4aS)-6c-[O2-(3,5,3'-trihydroxy-biphenyl-2-carbonyl)-beta-D-glucopyranosyloxy]-5t-vinyl-(4ar)-4,4a,5,6-tetrahydro-3H-pyrano[3,4-c]pyran-1-one Natural products OC1C(O)C(CO)OC(OC2C(C3C(C(OCC3)=O)=CO2)C=C)C1OC(=O)C1=C(O)C=C(O)C=C1C1=CC=CC(O)=C1 DBOVHQOUSDWAPQ-UHFFFAOYSA-N 0.000 claims description 86
- BZXINCMCFVKGKB-UHFFFAOYSA-N Amarogentin Natural products OCC1OC(OC2OC=C3C(CCOC3=O)C2C=C)C(OC(=O)c4cc(O)cc(O)c4c5cccc(O)c5)C(O)C1O BZXINCMCFVKGKB-UHFFFAOYSA-N 0.000 claims description 86
- 208000027418 Wounds and injury Diseases 0.000 claims description 20
- 230000009529 traumatic brain injury Effects 0.000 claims description 17
- 239000003292 glue Substances 0.000 claims description 14
- 239000003814 drug Substances 0.000 claims description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 229940079593 drug Drugs 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 239000003638 chemical reducing agent Substances 0.000 claims description 3
- 208000035475 disorder Diseases 0.000 claims description 2
- 238000001338 self-assembly Methods 0.000 claims description 2
- 239000003381 stabilizer Substances 0.000 claims description 2
- 238000002604 ultrasonography Methods 0.000 claims 1
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 4
- 230000000472 traumatic effect Effects 0.000 abstract description 4
- 230000004071 biological effect Effects 0.000 abstract 1
- 239000000178 monomer Substances 0.000 abstract 1
- 238000001132 ultrasonic dispersion Methods 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 16
- 230000006378 damage Effects 0.000 description 15
- 102000003777 Interleukin-1 beta Human genes 0.000 description 12
- 108090000193 Interleukin-1 beta Proteins 0.000 description 12
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 12
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 12
- 208000014674 injury Diseases 0.000 description 11
- 102000003814 Interleukin-10 Human genes 0.000 description 10
- 108090000174 Interleukin-10 Proteins 0.000 description 10
- 241000700159 Rattus Species 0.000 description 10
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- 239000006180 TBST buffer Substances 0.000 description 7
- 229930182470 glycoside Natural products 0.000 description 7
- 150000002338 glycosides Chemical class 0.000 description 7
- 244000018633 Prunus armeniaca Species 0.000 description 6
- 235000009827 Prunus armeniaca Nutrition 0.000 description 6
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 6
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 239000007928 intraperitoneal injection Substances 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 229910052710 silicon Inorganic materials 0.000 description 6
- 239000010703 silicon Substances 0.000 description 6
- 206010002091 Anaesthesia Diseases 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 230000037005 anaesthesia Effects 0.000 description 5
- 238000010171 animal model Methods 0.000 description 5
- 230000003247 decreasing effect Effects 0.000 description 5
- 238000005485 electric heating Methods 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000006872 improvement Effects 0.000 description 5
- 238000011068 loading method Methods 0.000 description 5
- 238000012453 sprague-dawley rat model Methods 0.000 description 5
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 230000003902 lesion Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 150000001408 amides Chemical class 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000009514 concussion Effects 0.000 description 3
- 210000003414 extremity Anatomy 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 230000003447 ipsilateral effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 3
- 230000036285 pathological change Effects 0.000 description 3
- 231100000915 pathological change Toxicity 0.000 description 3
- 230000004224 protection Effects 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000252506 Characiformes Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- KWYHDKDOAIKMQN-UHFFFAOYSA-N N,N,N',N'-tetramethylethylenediamine Chemical compound CN(C)CCN(C)C KWYHDKDOAIKMQN-UHFFFAOYSA-N 0.000 description 2
- 239000000020 Nitrocellulose Substances 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 208000006673 asthma Diseases 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 230000002490 cerebral effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 210000003128 head Anatomy 0.000 description 2
- LELOWRISYMNNSU-UHFFFAOYSA-N hydrogen cyanide Chemical compound N#C LELOWRISYMNNSU-UHFFFAOYSA-N 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 229920001220 nitrocellulos Polymers 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 230000000384 rearing effect Effects 0.000 description 2
- 230000011514 reflex Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 1
- XLSLFPQAPYONPW-WHUHBCJBSA-N (2s,3s,4s,5r,6r)-6-[(r)-cyano(phenyl)methoxy]-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound O1[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O[C@@H](C#N)C1=CC=CC=C1 XLSLFPQAPYONPW-WHUHBCJBSA-N 0.000 description 1
- 235000011437 Amygdalus communis Nutrition 0.000 description 1
- 244000144730 Amygdalus persica Species 0.000 description 1
- 206010003084 Areflexia Diseases 0.000 description 1
- RAFGELQLHMBRHD-VFYVRILKSA-N Bixin Natural products COC(=O)C=CC(=C/C=C/C(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C(=O)O)/C)C RAFGELQLHMBRHD-VFYVRILKSA-N 0.000 description 1
- 241000167854 Bourreria succulenta Species 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- XFXPMWWXUTWYJX-UHFFFAOYSA-N Cyanide Chemical compound N#[C-] XFXPMWWXUTWYJX-UHFFFAOYSA-N 0.000 description 1
- 206010014561 Emphysema Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 102000003815 Interleukin-11 Human genes 0.000 description 1
- 108090000177 Interleukin-11 Proteins 0.000 description 1
- UNGXBWFICNLIRX-UHFFFAOYSA-N Laetrile Natural products NCC(OC1OC(C(O)C(O)C1O)C(=O)O)c2ccccc2 UNGXBWFICNLIRX-UHFFFAOYSA-N 0.000 description 1
- 206010024229 Leprosy Diseases 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000220304 Prunus dulcis Species 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 235000020224 almond Nutrition 0.000 description 1
- RAFGELQLHMBRHD-UHFFFAOYSA-N alpha-Fuc-(1-2)-beta-Gal-(1-3)-(beta-GlcNAc-(1-6))-GalNAc-ol Natural products COC(=O)C=CC(C)=CC=CC(C)=CC=CC=C(C)C=CC=C(C)C=CC(O)=O RAFGELQLHMBRHD-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000001670 anatto Substances 0.000 description 1
- 235000012665 annatto Nutrition 0.000 description 1
- 230000000879 anti-atherosclerotic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000767 anti-ulcer Effects 0.000 description 1
- 229940030225 antihemorrhagics Drugs 0.000 description 1
- 210000000709 aorta Anatomy 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- RAFGELQLHMBRHD-SLEZCNMESA-N bixin Chemical compound COC(=O)\C=C\C(\C)=C/C=C/C(/C)=C/C=C/C=C(\C)/C=C/C=C(\C)/C=C/C(O)=O RAFGELQLHMBRHD-SLEZCNMESA-N 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 238000000861 blow drying Methods 0.000 description 1
- 230000037185 brain physiology Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000010523 cascade reaction Methods 0.000 description 1
- 235000019693 cherries Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 210000000613 ear canal Anatomy 0.000 description 1
- 230000005489 elastic deformation Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 210000003194 forelimb Anatomy 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 230000004914 glial activation Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000000025 haemostatic effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229940074383 interleukin-11 Drugs 0.000 description 1
- 230000000745 ion overload Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- NNICRUQPODTGRU-UHFFFAOYSA-N mandelonitrile Chemical compound N#CC(O)C1=CC=CC=C1 NNICRUQPODTGRU-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000004112 neuroprotection Effects 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000009023 proprioceptive sensation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000007026 protein scission Effects 0.000 description 1
- 208000005069 pulmonary fibrosis Diseases 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000518 rheometry Methods 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- HFFLGKNGCAIQMO-UHFFFAOYSA-N trichloroacetaldehyde Chemical compound ClC(Cl)(Cl)C=O HFFLGKNGCAIQMO-UHFFFAOYSA-N 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- 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
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Pain & Pain Management (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Rheumatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Medicinal Preparation (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention discloses a preparation method and application of tyrosine derived amygdalin-loaded hydrogel, wherein the preparation method comprises the steps of dissolving Fmoc-Tyr-OH solid powder by dimethyl sulfoxide, and obtaining Fmoc-Tyr-OH stock solution by ultrasonic dispersion; dissolving the amygdalin solution with ultrapure water to prepare amygdalin stock solution; and mixing the Fmoc-Tyr-OH stock solution and the amygdalin stock solution, shaking up, and standing to obtain the tyrosine derivative-loaded amygdalin hydrogel. The tyrosine derivative-delivered amygdalin hydrogel disclosed by the invention has an anti-inflammatory property, can solve the problem that an amygdalin monomer cannot directly act on a traumatic craniocerebral injury part, retains the biological activity of amygdalin, and has an obvious treatment effect when being applied to treatment of inflammatory reaction caused by traumatic craniocerebral injury.
Description
Technical field
The present invention relates to the preparation method and application that a kind of tyrosine-derived loads amarogentin hydrogel, belong to oversubscription
Sub- technical field of hydrogel.
Background technique
The amarogentin laetrile that is otherwise known as is a kind of cyanide, chemical molecular formula C20H27NO11, molecular weight
It is 457.42.Amarogentin is widely present in many plants, especially in the seed of rosaceous plant, such as almond, peach
Benevolence, cherry, plum etc..Amarogentin can hydrolyze and generate bixin and benzaldehyde cyanohydrin, under the action of glucuroide, most
Benzaldehyde and hydrogen cyanide are resolved into eventually.Much research shows that amarogentin has relieving cough and asthma, antiatherosclerosis, inhibition
Kidney region fibrosis, inhibit pulmonary fibrosis, anti-hyperbaric oxygen injury of lungs, immunosupress, immunological regulation, it is antitumor, anti-inflammatory and
The effect of antiulcer.Amarogentin be used to treat asthma, bronchitis, pulmonary emphysema, leprosy, colorectal cancer and leucoderma etc..
Because amarogentin can be decomposed into benzaldehyde, and benzaldehyde has analgesic effect, can be used to relieve pain.
Characters of traumatic brain injury (Traumatic brain injury, TBI) be the main lethal and disease that disables it
One, Characters of traumatic brain injury can lead to 50000 person/year of death.Characters of traumatic brain injury threatens the health of the mankind, and
Huge economic loss is caused to society and family, the above problem also promotes people to find better treatment method.
Caused by Characters of traumatic brain injury is leavened dough mainly due to primary injury and secondary lesion two.Primary injury
Craniocerebral injury caused by when referring to injury, mainly as mechanical external force is hit and caused by damage, this be can not carry out it is artificial
Intervene.Secondary lesion refers to a series of " waterfall type " pathologic, physiologics cascade occurred after the primary injury inside brain
Reaction, wherein inflammatory reaction is one of main pathological change.Characters of traumatic brain injury is mainly due to primary injury and secondary
Property damage two fermentation caused by.Craniocerebral injury caused by when primary injury refers to injury, mainly since mechanical external force is hit
It is damaged caused by and.Secondary lesion refers to a series of " waterfall type " pathology occurred after the primary injury inside brain
Physiology cascade reaction, wherein inflammatory reaction is one of main pathological change.Because primary injury be can not human intervention,
Therefore clinically main to pass through the pathologic process for intervening secondary lesion, to play neuroprotection.
Various factors not only follows self-growth after secondary craniocerebral injury, but also various factors is interweaved, phase interaction
With, collectively promote, ultimately form complicated damage network.Numerous studies discovery after craniocerebral injury can lead to inflammatory reaction, calcium
The pathological changes such as ion overload, excititoxic and oxidative stress.Wherein inflammatory reaction is that Characters of traumatic brain injury is latter
A key factor.It has been confirmed that the release of the excessive activation and proinflammatory factor of inflammation, example can be caused after Characters of traumatic brain injury
Such as IL 1-β (Interleukin 1 β, IL-1 β), tumor necrosis factor (Tumor necrosis factor, TNF-α)
Deng.The up-regulation of these inflammatory factors can increase blood-brain barrier permeability and lasting Glial Activation, these results
Can inflammatory reaction be further amplified again.And pertinent literature report passes through inflammatory reaction after intervention Characters of traumatic brain injury
It can promote the recovery of nervous function and play cerebral protection, wherein Science and JAMA is just clearly indicated that is created by intervening
The inflammatory reaction of wound property after craniocerebral injury can play cerebral protection.Therefore, pass through the inflammation after intervention Characters of traumatic brain injury
Reaction is an effective and feasible method and crucial therapy target.
Amarogentin drug has a good anti-inflammatory pharmacological action, but due to traditional Chinese medicine dosage form have in human body it is certain
Drug half-life, and Characters of traumatic brain injury position cannot be directly acted on, it is difficult to play corresponding curative effect, it is therefore desirable to
A kind of new mode improves its therapeutic effect.
Summary of the invention
In order to solve the above technical problems, the present invention provides the systems that a kind of tyrosine-derived loads amarogentin hydrogel
Preparation Method and application, the tyrosine-derived of production, which loads amarogentin hydrogel, can act on Characters of traumatic brain injury position,
With significant curative effect.
In order to achieve the above objectives, technical scheme is as follows: a kind of tyrosine-derived loading amarogentin hydrogel
Preparation method, comprising the following steps:
Step 1, Fmoc-Tyr-OH stock solution is configured；With dmso solution Fmoc-Tyr-OH solid powder, surpass
Sound disperses to obtain Fmoc-Tyr-OH stock solution；
Step 2, amarogentin stock solution is configured；Amarogentin solution is dissolved with ultrapure water, is made into amarogentin deposit
Solution；
Step 3, hydrogel is prepared；The Fmoc-Tyr-OH stock solution and amarogentin stock solution are mixed and shaken
It is even, it stands and obtains tyrosine derivative loading amarogentin hydrogel.
As the improvement of above-mentioned preparation method, dmso solution Fmoc-Tyr-OH solid powder is used in step 1, is surpassed
The time of sound dispersion is 0.5~1 minute, and the concentration of acquired Fmoc-Tyr-OH stock solution is 50~100mg/mL.
As the improvement of above-mentioned preparation method, in step 2, the concentration of the amarogentin stock solution is 1~16mg/
mL。
As the improvement of above-mentioned preparation method, in step 3, takes the Fmoc-Tyr-OH stock solution of 30~50 μ L to be added and wash
Only in the screw socket bottle of dry 5.0mL, 950~970 μ L amarogentin stock solutions is rapidly joined, are shaken up rapidly, it is then quiet
It sets, the tyrosine derivative loads amarogentin hydrogel and formed.
As the improvement of above-mentioned preparation method, under conditions of not adding reducing agent and stabilizer, the tyrosine-derived
It is the clear obtained by Fmoc-Tyr-OH gelator by non-covalent bond self assembly that object, which loads amarogentin hydrogel,
Hydrogel.
A kind of tyrosine-derived of above-mentioned preparation method production loads amarogentin hydrogel and is treating traumatic head trauma
Hurt the application in the drug of disease.
A kind of tyrosine-derived of above-mentioned preparation method production load amarogentin hydrogel mitigate or treat it is traumatic
Application in craniocerebral injury Disorders Inflammation disease medicament.
Through the above technical solutions, the beneficial effect of technical solution of the present invention is:
1) tyrosine derivative prepared by the present invention loads amarogentin hydrogel, and good biocompatibility is degradable.
2) supramolecular hydrogel prepared by the present invention is the hydrogel of clear, has suitable hardness, elasticity and glues
Property, it is easy to inject.
3) without adding any reducing agent and dispersing agent in preparation process of the present invention, supermolecule can be formed by standing
Hydrogel, it is environmentally protective.
4) preparation method of the present invention is simple, low in cost, can be commercialized, and can be adapted for applying.
5) supramolecular hydrogel prepared by the present invention has good anti-inflammatory effect, after capable of inhibiting Characters of traumatic brain injury
Inflammatory reaction.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, below will to embodiment or
Attached drawing needed to be used in the description of the prior art is briefly described, it should be apparent that, the accompanying drawings in the following description is this hair
Bright some embodiments for those of ordinary skill in the art without creative efforts, can be with
It obtains other drawings based on these drawings.
Fig. 1 is scanning electron microscope (SEM) figure of tyrosine derivative supramolecular hydrogel；
Fig. 2 is scanning electron microscope (SEM) figure of amarogentin solution；
Fig. 3 is the scanning electron microscopy that tyrosine derivative prepared by the embodiment of the present invention 1 loads amarogentin hydrogel
Mirror (SEM) figure.
Fig. 4 is the digital photograph figure that tyrosine derivative prepared by the embodiment of the present invention 1 loads amarogentin hydrogel.
Fig. 5 is Fourier's infrared spectrum that tyrosine derivative loads amarogentin hydrogel in the embodiment of the present invention 2.
Fig. 6 is that the tyrosine derivative in the embodiment of the present invention 3 loads amarogentin hydrogel rheogram.
Fig. 7 is that tyrosine derivative loads amarogentin hydrogel and tyrosine hydrogel to wound in the embodiment of the present invention 4
The influence of the Neuroscore of rat in the craniocerebral injury of wound property.
Fig. 8 is that tyrosine derivative loads amarogentin hydrogel and tyrosine hydrogel to wound in the embodiment of the present invention 4
The influence of interleukin-11 β (IL-1 β) in the craniocerebral injury of wound property.
Fig. 9 is that tyrosine derivative loads amarogentin hydrogel and tyrosine hydrogel to wound in the embodiment of the present invention 4
The influence of tumor necrosis factor α (TNF-α) inflammatory factor level in the craniocerebral injury of wound property.
Figure 10 is that tyrosine derivative loads amarogentin hydrogel and tyrosine hydrogel pair in the embodiment of the present invention 4
The influence of IL-10 (IL-10) inflammatory factor level in Characters of traumatic brain injury.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical solution in the embodiment of the present invention is clearly and completely retouched
It states, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the present invention
In embodiment, every other implementation obtained by those of ordinary skill in the art without making creative efforts
Example, shall fall within the protection scope of the present invention.
Embodiment 1
The Fmoc-Tyr-OH solid powder for weighing 5.0mg/mL, is dissolved in 100 μ L dimethyl sulfoxides, and ultrasonic disperse 30 seconds,
So that it is completely dissolved, the Fmoc-Tyr-OH stock solution that preparation concentration is 50mg/mL；Weigh the amarogentin of 5.0mg in
In centrifuge tube, the ultrapure water that 1.0mL is added is dissolved, the amarogentin stock solution that preparation concentration is 5mg/mL.
It takes the Fmoc-Tyr-OH stock solution of 50 μ L to be added in the 5.0mL screw socket bottle of clean dry, 970 μ L hardship apricots is added
Benevolence glycosides stock solution, rapid oscillation shake up, and stand the tyrosine derivative loading semen armeniacae amarae for forming clear for 3 minutes
The hydrogel of glycosides.
The experimental procedure of scanning electron microscope:
With Piranha solution, (concentrated sulfuric acid (v): hydrogen peroxide (v)=7: then ethyl alcohol is used in 3) ultrasonic cleaning silicon wafer 15 minutes
Solution is cleaned by ultrasonic 15 minutes, is finally cleaned 15 minutes, is then taken out silicon wafer from Piranha solution, with two with ultrapure water
Secondary water rinses silicon wafer, then with nitrogen by silicon wafer blow-drying.It takes 10 μ L amarogentin hydrogels to be placed on silicon wafer with liquid-transfering gun, puts
Enter -4 DEG C of refrigerator freezings, after the sample of freezing is put into vacuum freeze dryer, will wherein moisture extract.Before carrying out SEM test
Sample will be prepared and be put into drier preservation.When carrying out SEM test, silicon wafer is put into sample stage, then sample is sprayed
Gold processing, is put into instrument, observes under vacuum conditions.
Fig. 1 is tyrosine derivative supramolecular hydrogel scanning electron microscope diagram, and Fig. 2 is the scanning of amarogentin solution
Electron microscope picture, Fig. 3 are that tyrosine derivative loads amarogentin supramolecular hydrogel electron-microscope scanning result.As shown,
Tyrosine derivative can form reticular structure, as shown in Figure 1, tyrosine derivative can load amarogentin, such as Fig. 3 institute
Show.Fig. 4 is the digital photograph figure that tyrosine derivative prepared by the embodiment of the present invention 1 loads amarogentin hydrogel
Embodiment 2
The Fmoc-Tyr-OH solid powder for weighing 5.0mg/mL, is dissolved in 100 μ L dimethyl sulfoxides, and ultrasonic disperse 30 seconds,
So that it is completely dissolved, the Fmoc-Tyr-OH stock solution that preparation concentration is 100mg/mL；Weigh the amarogentin of 16.0mg
In centrifuge tube, the ultrapure water that 1.0mL is added is dissolved, the amarogentin stock solution that preparation concentration is 16mg/mL.
It takes the Fmoc-Tyr-OH stock solution of 30 μ L to be added in the 5.0mL screw socket bottle of clean dry, 950 μ L hardship apricots is added
Benevolence glycosides stock solution, concussion shakes up rapidly, and stands the water for forming the tyrosine loading amarogentin of clear for 3 minutes
Gel.
Fourier's infrared test experimental procedure: sample after dry 12 hours, is weighed into 0.2g bromine in freeze drier
Change potassium powder and 2mg sample is then added, is put into grinding in mortar, the two is mixed well, then mixture is put into mold
It is pressed into sheet, carries out infrared test.Fig. 5 is that tyrosine derivative loads amarogentin and the Fourier of amarogentin powder is red
Outer spectrogram.Test result is as shown in Fig. 5, from available in figure: the characteristic absorption of proteins and peptides in infrared spectroscopy
Band is defined as amino acid A, B and I-VII.I (1600-1690cm of amide-1) and II (1480-1575cm of amide-1) absorption
Band is frequently used to analysis Secondary structure.Distinguish in the absorption peak of amide I, beta sheet, alpha-helix and random coils
Close to 1630cm-1、1655cm-1、1645cm-1.Fmoc-Tyr-OH-Amygdalin its in 1643cm-1Place has an amide I to inhale
Peak is received, amarogentin (Amygdalin) is in 1642cm-1There is an absorption peak at place.Amide II is due to the flexible concussion of C-N and N-
H bending is derived.Both in 1481cm-1There is absorption peak at place.These are the result shows that Fmoc- is added in amarogentin drug
The change of amarogentin structure is not caused in Tyr-OH hydrogel.
Embodiment 3
The Fmoc-Tyr-OH solid powder for weighing 5.0mg/mL, is dissolved in 100 μ L dimethyl sulfoxides, and ultrasonic disperse 60 seconds,
So that it is completely dissolved, the Fmoc-Tyr-OH stock solution that preparation concentration is 75mg/mL；Weigh the amarogentin of 1.0mg in
In centrifuge tube, the ultrapure water that 1.0mL is added is dissolved, the amarogentin stock solution that preparation concentration is 1mg/mL.
It takes the Fmoc-Tyr-OH stock solution of 40 μ L to be added in the 5.0mL screw socket bottle of clean dry, 960 μ L hardship apricots is added
Benevolence glycosides stock solution, concussion shakes up rapidly, and stands the tyrosine loading amarogentin water-setting for forming clear for 3 minutes
Glue.
Rheometer test experimental procedure: testing its rheological behavior using AR2000 rheology instrument, the use of diameter is 50mm
Parallel-plate, gap setting 0.2mm, test condition: temperature be 25 DEG C, intensity 2%, frequency be 0.1-100Hz.
Test results are shown in figure 6, while determining Fmoc-Tyr-OH supramolecular hydrogel and Fmoc-Tyr-OH-
Storage modulus (G ') loss modulus (G ") of Amygdalin supramolecular hydrogel, wherein storage modulus (G ') indicate material because
Elastic deformation and the energy stored, loss modulus (G ") indicate the viscous deformation due to material and the energy that consumes, and consume angle is just
The ratio that value (tan θ) is loss modulus (G ") and storage modulus (G ') is cut, the viscoelastic ratio of material is used to represent.One
As gel rubber system in G ' > G ", i.e. tan θ < 1.As shown in fig. 6, the value of G ' is much larger than G " in the frequency range of scanning,
And the relatively simple Fmoc-Tyr-OH supramolecular hydrogel of its G ' after amarogentin is added in Fmoc-Tyr-OH supramolecular hydrogel
Glue increases.According to the above results, amarogentin, which is loaded into Fmoc-Tyr-OH supramolecular hydrogel, will not influence its property.
Embodiment 4
The 30 μ l of Fmoc-Tyr-OH stock solution in above-described embodiment 1 is taken, the 5.0ml screw socket bottle of clean dry is added
In, 970 μ l 8mg/ml amarogentin stock solution identical with 1 preparation method of embodiment is rapidly joined, is shaken up rapidly, is stood
The tyrosine for forming clear loads amarogentin hydrogel.
Experimental animal:
Sprague-Dawley (SD) rat of cleaning grade male is chosen in this experiment, and all experimental animals are bought in
Southern university experimental animal portion, credit number: SCXK (Hunan) 2016-0002, weight: 200-250g.During experimental animal has passed through
Southern university animal Ethics Committee approval, and experimentation follows " the guiding opinion about kind treatment experimental animal " (2006
Version).All animal sub-cage rearings are in Animal Experimental SPF grades of animal house of Central South University: room temperature maintains (22-25) DEG C, environment
Humidity maintains (50 ± 10) %, and daily 12 hours circulation lights shine.
The preparation of Characters of traumatic brain injury model:
Traumatic brain injury model is replicated using USA TBI 0310Head Impactor, the specific method is as follows: with 5% hydration
SD rat head is fixed on craniocerebral injury instrument, preserved skin is strictly abided by by chloral 4mL/kg intraperitoneal injection after anaesthetizing SD rat
It follows sterile working, after Iodophor routine disinfection three times, cuts off skin, exposure skull, in right side bregma with after with sterile cut from center
The bone window that a diameter is 5mm is opened in center with dental burr between fontanel, bone window rat will be ended and be fixed on craniocerebral injury instrument
On device, experiment parameter is set are as follows: depth 5mm, speed 6mac/s, residence time 2s after strike, suture skin
Skin simultaneously sterilizes, and animal is put on thermostatic electric heating blanket and is kept warm wait revive, sub-cage rearing after revival.
Experimental animal grouping:
Male SD rat is randomly divided into 4 groups, sham-operation group (Sham), craniocerebral injury group (TBI+PBS), craniocerebral injury+
Tyrosine derivative hydrogel group (TBI+Fmoc-Tyr-OH), craniocerebral injury+tyrosine load amarogentin hydrogel group
(TBI+8mg/mL Fmoc-Tyr-OH-Amygdalin), concrete scheme is as follows:
Sham-operation group (Sham): after operation for 24 hours, after intraperitoneal injection 5% chloraldurate 4mL/kg anesthesia, at the window that opens seam
20 μ LPBS (with 4 μ L/min speed), skin suture are injected, and is sterilized, animal is put into thermostatic electric heating blanket and is kept warm, wait revive.
Craniocerebral injury group (TBI+PBS): it after operation for 24 hours, after intraperitoneal injection 5% chloraldurate 4mL/kg anesthesia, is opening seam
20 μ L PBS (with 4 μ L/min speed) are injected at window, skin suture, and sterilize, animal is put into thermostatic electric heating blanket and is kept warm, to
Revival.
Craniocerebral injury+tyrosine derivative hydrogel group (TBI+Fmoc-Tyr-OH): 24 h after operation, intraperitoneal injection 5%
After chloraldurate 4mL/kg anesthesia, 20 μ L-Tyr derivative hydrogels (with 4 μ L/min speed) are injected at the window that opens seam, seam
Skin is closed, and is sterilized, animal is put into thermostatic electric heating blanket and is kept warm, wait revive.
Craniocerebral injury+8mg/mL tyrosine derivative loads amarogentin hydrogel group (Fmoc-Tyr-OH-
Amygdalin): after operation for 24 hours, after intraperitoneal injection 5% chloraldurate 4mL/kg anesthesia, 20 μ L8mg/ are injected at the window that opens seam
ML amarogentin hydrogel (with 4 μ L/min speed), skin suture, and sterilize, animal is put into thermostatic electric heating blanket and is kept warm, to
Revival.
Neuroscore:
| Project | Score |
| Best result | 18 |
| Dynamic test | |
| Tail rat is held to gradually rise | 3 |
| 1 forelimb buckling | |
| 1 hind leg buckling | |
| 1 upper lift maintains in 30s away from 10 ° of vertical axis > | |
| Rat is placed in ground (normally=0；Maximum value=3) | 3 |
| 0 normally creeps | |
| 1 can not rectilinear creeping | |
| 2 turn-take along Ipsilateral | |
| 3 Ipsilaterals are fallen | |
| Sensory test | 2 |
| 1 location aware test (vision and tactile test) | |
| 2 proprioceptions test (bathesthesia, opposite table edge pawl son to stimulate limb muscle) | |
| Beam balance test (normal=0；Maximum value=6) | 6 |
| It can be balanced under 0 static posture | |
| 1 grasps beam side | |
| 2 embrace beam, have a limbs to fall | |
| 3 embrace beam, have two limbs to fall or rotate (> 60s) on beam | |
| 4 attempt to keep balance on beam, but fall (> 40s) | |
| 5 attempt to keep balance on beam, but fall (> 20s) | |
| 6 fall, but do not attempt to keep balance or grasp beam (< 20s) | |
| Areflexia or abnormal operation | 4 |
| 1 auricle reflex (is shaken the head) when touching ear canal | |
| 1 corneal reflection (blink when touching cornea with cotton silk) | |
| 1 Moro embrace reflex (motor reaction caused by noise) | |
| 1 grasping, myoclonia, myodystony |
Note: score is higher, and the degree of injury that represents is higher, and 1 point of representative has no ability to test or tested reflection lacks
It loses.
The expression of Western blotting measurement Damage of Rats side cortex IL-1 β, TNF-α, IL-10
(1) sample preparation: with chloraldurate with 4mL/kg intraperitoneal injection of anesthesia after, rapidly expose heart, with catheter needle from
Left ventricular apex is inserted into aorta direction, breaks right auricle of heart with ice saline infusions with haemostatic clamp fixed needle, until right
Until auricle flows out colorless clear liquid.Then broken end takes brain, takes Ipsilateral cortex respectively, is packaged in cryopreservation tube.Clip group
Knit, rinse tissue with ice PBS, be added 200uL RIPA lysate in homogenizer repeatedly tissue abrasion until invisible tissue
Block.On ice, protein cleavage 10 minutes are allowed, after cracking, centrifuge is centrifuged 15 points with 12000rpm/min at 4 DEG C
It is spare to wash away supernatant for clock.
(2) protein concentration detects: suitable BCA working solution is prepared with the BCA reagent A of 50:1 and BCA reagent B, it is sufficiently mixed
It is even.It is completely dissolved protein standard substance, concentration 2mg/mL is distinguished standard items by 0 μ L, 1 μ L, 2 μ L, 3 μ L, 4 μ L, 5 μ L, 6 μ L
It is added in the standard sample wells of 96 orifice plates, the solution for dilution standard product is added to supply to 20 μ L.Standby sample is added to 96 holes
Plate adds solution for dilution standard product to 20 μ L.
4. 200 μ L BCA working solutions are added in each hole, 30min is placed in 37 DEG C.The measuring point absorbance at 562nm,
And protein concentration is calculated according to standard curve.
(3) electrophoresis: 10% separation gel is prepared, shaking up immediately after addition TEMED can encapsulating.After encapsulating, isopropanol is used
Sealing.Isopropanol on glue is gone after solidifying glue sufficiently, with filter paper suck dry moisture.4.8% concentration glue is prepared, is added
Concentration glue is shaken up immediately after TEMED, configured concentration glue is added.Adjusting each group applied sample amount according to protein quantification result makes
Protein content is consistent in sample, and 5*loading buffer is added and mixes, boiling water boiling 5min makes albuminous degeneration, is put into ice chest
Quickly cooling.According to protein quantification as a result, maker is clicked and entered in the first hole, other every holes are added each group sample, start electrophoresis.Electrophoresis
Concentrate glue voltage is 80V, and separation gel electrophoretic voltage is 120V.Electrophoresis is terminated when bromophenol blue electrophoresis to glue bottom.
(4) transferring film: glue IL-1 β, TNF-α, IL-10 are cut respectively.Prepare 6 and an equal amount of filter paper of cut glue and 1
Nitrocellulose filter, nitrocellulose filter and filter paper are put into togerther in transferring film buffer, until being impregnated with completely.According to filter paper, film,
Glue, filter paper sequence successively put well, intermediate bubble is drained.Instrument is covered, is powered on, transferring film voltage is 300mA, time
For 45min~1h.After transferring film, takes the film out to be put into 1*TBST and wash 1 time, time 5min.Film, detection are contaminated with Ponceaux
The efficiency of albumen transferring film.Ponceaux is cleaned with 1*TBST.
(5) it closes: preparing 5% skimmed milk power with 1*TBST, after film is immersed, be placed at room temperature for 1.5h.
(6) primary antibody is incubated for: primary antibody being diluted (table 3-2) according to a certain percentage with 1*TBST, film is incubated together with primary antibody
It educates, 4 DEG C overnight.Incubation terminates, and 1*TBST is washed 3 times, each 15min.
Each antibody uses in table 3-2 western blot
(7) secondary antibody is incubated for: the secondary antibody (Proteintech) marked with 1*TBST dilution HRP, rabbit-anti (R) dilution ratio 1:
6000, mouse resists (M) dilution ratio 1:5000, and the secondary antibody after dilution is incubated for 90min with film jointly.Incubation terminates, and 1*TBST is washed
3 times, each 15min.
(8) colour developing exposure: being incubated for 3min using ECL chemical luminescence for liquid (Thermo) and film, exhausts liquid with blotting paper,
Film is wrapped up into hybond membrane with preservative film, in magazine with several minutes of X exposure；Development is rinsed.
Experimental result is shown in Fig. 7, Fig. 8, Fig. 9 and Figure 10 respectively.
Fig. 7 is that tyrosine derivative loads amarogentin supramolecular hydrogel and tyrosine supramolecular hydrogel to wound
The influence of property after craniocerebral injury rat nerve function score.As can be seen from Figure 7: compared with sham-operation group, model group nerve function
It can score and dramatically increase (P < 0.01)；Compared with model group, tyrosine derivative supramolecular hydrogel Neuroscore without
Notable difference (P > 0.05), tyrosine derivative, which loads amarogentin supramolecular hydrogel group Neuroscore, reduces (P <
0.05)；Tyrosine derivative loads amarogentin hydrogel group compared with tyrosine derivative group, and Neuroscore is obvious
Decline (P < 0.05), the improvement for illustrating its nervous function is tyrosine derivative hydrogel by discharging amarogentin drug
It plays a role.
Fig. 8 is that tyrosine derivative loads amarogentin and tyrosine derivative supramolecular hydrogel to the shadow of IL-1 β
It rings, as can be seen from Figure 6: compared with sham-operation group, the horizontal significant raising (P < 0.01) of model group IL-1 β；With model group ratio
Compared with tyrosine derivative hydrogel group IL-1 β expression changes unobvious (P > 0.05), and tyrosine derivative loads bitter apricot
Benevolence glycosides hydrogel group IL-1 β expression is remarkably decreased (P < 0.01), and tyrosine derivative loads amarogentin hydrogel group
Compared with tyrosine derivative hydrogel group, IL-1 β expression is decreased obviously (P < 0.05), illustrates that it mainly passes through and releases
It puts amarogentin and inhibits IL-1 β expression.
Fig. 9 is that tyrosine derivative loads amarogentin and tyrosine derivative supramolecular hydrogel to the shadow of TNF-α
It rings, as can be seen from Figure 9: compared with sham-operation group, the horizontal significant raising (P < 0.01) of model group TNF-α；With model group ratio
Compared with tyrosine derivative hydrogel group TNF-α expression changes unobvious (P > 0.05), and tyrosine derivative loads bitter apricot
Benevolence glycosides hydrogel group TNF-α expression is remarkably decreased (P < 0.01), and tyrosine derivative loads amarogentin hydrogel group
Compared with tyrosine derivative hydrogel group, IL-1 β expression is decreased obviously (P < 0.05), illustrates that it mainly passes through and releases
It puts amarogentin and inhibits TNF-α expression.
Figure 10 is that tyrosine derivative loads amarogentin and tyrosine derivative supramolecular hydrogel to the shadow of IL-10
It rings, as can be seen from Figure 7: compared with sham-operation group, model group IL-10 level is remarkably decreased (P < 0.01)；With model group ratio
Compared with tyrosine derivative hydrogel group IL-10 expression changes unobvious (P > 0.05), and tyrosine derivative loads bitter apricot
Benevolence glycosides hydrogel group IL-10 expression significantly increases (P < 0.01), and tyrosine derivative loads amarogentin hydrogel group
Compared with tyrosine derivative hydrogel group, IL-10 expression is significantly raised (P < 0.05), illustrates that it mainly passes through and releases
It puts amarogentin and promotes IL-10 expression.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, defined herein
General Principle can realize in other embodiments without departing from the spirit or scope of the present invention.Therefore, originally
Invention is not intended to be limited to the embodiments shown herein, and is to fit to special with principles disclosed herein and novelty
The consistent widest scope of point.
Claims (7)
1. the preparation method that a kind of tyrosine-derived loads amarogentin hydrogel, which comprises the following steps:
Step 1, Fmoc-Tyr-OH stock solution is configured；With dmso solution Fmoc-Tyr-OH solid powder, ultrasound point
It dissipates and obtains Fmoc-Tyr-OH stock solution；
Step 2, amarogentin stock solution is configured；Amarogentin solution is dissolved with ultrapure water, is made into amarogentin stock solution；
Step 3, hydrogel is prepared；The Fmoc-Tyr-OH stock solution and amarogentin stock solution are mixed and shaken up, is stood
It obtains tyrosine derivative and loads amarogentin hydrogel.
2. the preparation method that tyrosine-derived according to claim 1 loads amarogentin hydrogel, which is characterized in that step
Dmso solution Fmoc-Tyr-OH solid powder is used in rapid 1, the time of ultrasonic disperse is 0.5~1 minute, acquired
The concentration of Fmoc-Tyr-OH stock solution is 50~100mg/mL.
3. the preparation method that tyrosine-derived according to claim 2 loads amarogentin hydrogel, which is characterized in that step
In rapid 2, the concentration of the amarogentin stock solution is 1~16mg/mL.
4. the preparation method that tyrosine-derived according to claim 1 loads amarogentin hydrogel, which is characterized in that step
In rapid 3, in the screw socket bottle for the 5.0mL for taking the Fmoc-Tyr-OH stock solution of 30~50 μ L that clean dry is added, rapidly join
950~970 μ L amarogentin stock solutions, shake up rapidly, are then allowed to stand, and the tyrosine derivative loads amarogentin water-setting
Glue is formed.
5. tyrosine-derived according to claim 1 to 4 loads the preparation method of amarogentin hydrogel, feature
It is, under conditions of not adding reducing agent and stabilizer, the tyrosine derivative loads amarogentin hydrogel and serves as reasons
The hydrogel for the clear that Fmoc-Tyr-OH gelator is obtained by non-covalent bond self assembly.
6. a kind of tyrosine-derived of the production of preparation method described in claim 1-4 loads amarogentin hydrogel and creates in treatment
Application in the drug of wound property craniocerebral injury disease.
7. preparation method described in a kind of claim 1-4 production tyrosine-derived load amarogentin hydrogel mitigate or
Treat the application in Characters of traumatic brain injury Disorders Inflammation disease medicament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810996854.8A CN109125249B (en) | 2018-08-29 | 2018-08-29 | Preparation method and application of tyrosine-derived amygdalin-loaded hydrogel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810996854.8A CN109125249B (en) | 2018-08-29 | 2018-08-29 | Preparation method and application of tyrosine-derived amygdalin-loaded hydrogel |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109125249A true CN109125249A (en) | 2019-01-04 |
| CN109125249B CN109125249B (en) | 2021-10-26 |
Family
ID=64829200
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810996854.8A Expired - Fee Related CN109125249B (en) | 2018-08-29 | 2018-08-29 | Preparation method and application of tyrosine-derived amygdalin-loaded hydrogel |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109125249B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111096961A (en) * | 2020-02-18 | 2020-05-05 | 苏州大学 | Application of erythrosin in preparation of medicine for treating renal interstitial fibrosis |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101431983A (en) * | 2004-09-28 | 2009-05-13 | 香港科技大学 | Multifunctional supramolecular hydrogels as biomaterials |
| CN102133175A (en) * | 2011-03-09 | 2011-07-27 | 天津市南开医院 | Amygdalin gel and preparation method and medicinal application thereof |
-
2018
- 2018-08-29 CN CN201810996854.8A patent/CN109125249B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101431983A (en) * | 2004-09-28 | 2009-05-13 | 香港科技大学 | Multifunctional supramolecular hydrogels as biomaterials |
| CN102133175A (en) * | 2011-03-09 | 2011-07-27 | 天津市南开医院 | Amygdalin gel and preparation method and medicinal application thereof |
Non-Patent Citations (2)
| Title |
|---|
| CHUANBIN YANG ET AL.: "Bioactivity-Guided Fractionation Identifies Amygdalin as a Potent Neurotrophic Agent from Herbal Medicine Semen Persicae Extract", 《BIOMED RESEARCH INTERNATIONAL》 * |
| WATHSALA LIYANAGE AND BRADLEY L. NILSSON: "Substituent Effects on the Self-Assembly/Coassembly and Hydrogelation of Phenylalanine Derivatives", 《LANGMUIR》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111096961A (en) * | 2020-02-18 | 2020-05-05 | 苏州大学 | Application of erythrosin in preparation of medicine for treating renal interstitial fibrosis |
| CN111096961B (en) * | 2020-02-18 | 2022-05-13 | 苏州大学 | Application of bixin in preparing medicine for treating renal interstitial fibrosis |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109125249B (en) | 2021-10-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102600496B (en) | I, II and III medical equipment products acting on skin, mucosa and wound for inactivating virus | |
| WO2006032091A2 (en) | Herbal composition | |
| KR20170005115A (en) | Composition of natural vitamin c and fish scale collagen peptide and preparation method thereof | |
| WO2021128996A1 (en) | Natural plant bacteriostatic ointment containing artemisia argyi extract and preparation method therefor | |
| CN110787114A (en) | Foam shampoo capable of removing dandruff, relieving itching and controlling oil and preparation method thereof | |
| JP2018521992A5 (en) | ||
| CN106420485A (en) | Oligopeptide skin smoothing rejuvenating powder | |
| CN107519236A (en) | A kind of topical agent for treating onychomycosis | |
| CN109125249A (en) | Preparation method and application of tyrosine-derived amygdalin-loaded hydrogel | |
| JP2018048195A (en) | Homeopathic therapeutic method and composition | |
| CN110151981A (en) | A kind of composition and preparation method thereof repaired vagina and treat vaginitis | |
| CN116115707A (en) | Concentrated foam medicated bath liquid for preventing and treating cow hoof rot and preparation method thereof | |
| CN110141678A (en) | A kind of alginates physical therapy liquid dressing and preparation method thereof | |
| US9364510B2 (en) | Botanical composition and methods of manufacture and use | |
| RU2530589C1 (en) | Agent for treating septic wounds, method for preparing it and method of treating septic wounds | |
| CN103417555B (en) | Cerebroside compound is preparing the application in analgesic | |
| RU2757497C1 (en) | Method for treating necrobacteriosis of cloven-hoofed animals | |
| CN105106940A (en) | Collagen composition for treating acne and collagen patch | |
| CN109700876A (en) | A kind of oxidation resistant particle is sufficient to paste pack and preparation method thereof | |
| Sen et al. | Influence of Cinnamomum camphora ethanolic extract on Biophysical and Biochemical Parameters of Cutaneous Wounds in Rats | |
| CN115554386B (en) | Application of gastrodia elata polypeptide in preparing external product for inhibiting or killing trichophyton | |
| RU2740450C1 (en) | Dental gel with adaptogen for treating periodontal inflammatory diseases | |
| RU2132197C1 (en) | Agent possessing antitumor and immunomodulating effect | |
| KR101922488B1 (en) | A composition for treating and preventing hair loss containing pozzolan, Sappan wood extract and siegesbockia herb extract | |
| Shetty et al. | Evaluation and comparison of buffering capacity of apamarga and phalasha ksharas |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20211026 |