CN107158406A - The glycosylated SiO of Glucomannan2Nano particle and its preparation method and application - Google Patents
The glycosylated SiO of Glucomannan2Nano particle and its preparation method and application Download PDFInfo
- Publication number
- CN107158406A CN107158406A CN201710424892.1A CN201710424892A CN107158406A CN 107158406 A CN107158406 A CN 107158406A CN 201710424892 A CN201710424892 A CN 201710424892A CN 107158406 A CN107158406 A CN 107158406A
- Authority
- CN
- China
- Prior art keywords
- sio
- glucomannan
- particle
- glycosylated
- cdi
- 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
- 239000002245 particle Substances 0.000 title claims abstract description 45
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 73
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 71
- 229910052681 coesite Inorganic materials 0.000 claims abstract description 69
- 229910052906 cristobalite Inorganic materials 0.000 claims abstract description 69
- 229910052682 stishovite Inorganic materials 0.000 claims abstract description 69
- 229910052905 tridymite Inorganic materials 0.000 claims abstract description 69
- 239000002105 nanoparticle Substances 0.000 claims abstract description 41
- 150000004676 glycans Chemical class 0.000 claims abstract description 37
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 35
- 239000005017 polysaccharide Substances 0.000 claims abstract description 35
- 238000005576 amination reaction Methods 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 23
- 230000029663 wound healing Effects 0.000 claims abstract description 20
- 241001313855 Bletilla Species 0.000 claims abstract description 19
- 206010061218 Inflammation Diseases 0.000 claims abstract description 14
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 14
- 230000004054 inflammatory process Effects 0.000 claims abstract description 14
- 239000003814 drug Substances 0.000 claims abstract description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 54
- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 claims description 51
- 229920002581 Glucomannan Polymers 0.000 claims description 51
- 229940046240 glucomannan Drugs 0.000 claims description 51
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 32
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 18
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 13
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims description 12
- 208000007107 Stomach Ulcer Diseases 0.000 claims description 12
- 230000004913 activation Effects 0.000 claims description 12
- 201000005917 gastric ulcer Diseases 0.000 claims description 12
- 201000010099 disease Diseases 0.000 claims description 11
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 claims description 10
- 230000004048 modification Effects 0.000 claims description 10
- 238000012986 modification Methods 0.000 claims description 10
- 238000011282 treatment Methods 0.000 claims description 9
- 230000009514 concussion Effects 0.000 claims description 8
- 125000002883 imidazolyl group Chemical group 0.000 claims description 8
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims description 7
- 206010009887 colitis Diseases 0.000 claims description 7
- 230000013595 glycosylation Effects 0.000 claims description 6
- 238000006206 glycosylation reaction Methods 0.000 claims description 6
- 238000010438 heat treatment Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 3
- 208000007882 Gastritis Diseases 0.000 claims description 2
- 206010003246 arthritis Diseases 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 208000023652 chronic gastritis Diseases 0.000 claims description 2
- 238000002242 deionisation method Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 150000003851 azoles Chemical class 0.000 claims 2
- 210000002540 macrophage Anatomy 0.000 abstract description 15
- 230000017423 tissue regeneration Effects 0.000 abstract description 8
- 210000002784 stomach Anatomy 0.000 abstract description 7
- 230000004071 biological effect Effects 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 4
- 230000008929 regeneration Effects 0.000 abstract description 4
- 238000011069 regeneration method Methods 0.000 abstract description 4
- 230000004968 inflammatory condition Effects 0.000 abstract description 3
- 150000002704 mannoses Chemical class 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 230000001105 regulatory effect Effects 0.000 abstract description 3
- 229910004298 SiO 2 Inorganic materials 0.000 abstract description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 abstract description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 abstract description 2
- 230000001133 acceleration Effects 0.000 abstract description 2
- 230000001276 controlling effect Effects 0.000 abstract description 2
- 210000000936 intestine Anatomy 0.000 abstract description 2
- 230000035772 mutation Effects 0.000 abstract description 2
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 abstract 1
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 abstract 1
- 230000003213 activating effect Effects 0.000 abstract 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 36
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 25
- NHJVRSWLHSJWIN-UHFFFAOYSA-N 2,4,6-trinitrobenzenesulfonic acid Chemical compound OS(=O)(=O)C1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O NHJVRSWLHSJWIN-UHFFFAOYSA-N 0.000 description 20
- 239000000243 solution Substances 0.000 description 20
- 206010052428 Wound Diseases 0.000 description 14
- 208000027418 Wounds and injury Diseases 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 230000009266 disease activity Effects 0.000 description 10
- 230000002496 gastric effect Effects 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 230000002757 inflammatory effect Effects 0.000 description 9
- 238000001802 infusion Methods 0.000 description 9
- 230000008859 change Effects 0.000 description 8
- 210000001072 colon Anatomy 0.000 description 7
- 230000035876 healing Effects 0.000 description 6
- 239000002504 physiological saline solution Substances 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 208000024891 symptom Diseases 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 230000001225 therapeutic effect Effects 0.000 description 5
- 241001313857 Bletilla striata Species 0.000 description 4
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 4
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 235000013339 cereals Nutrition 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 150000004804 polysaccharides Polymers 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 208000032843 Hemorrhage Diseases 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 229910002808 Si–O–Si Inorganic materials 0.000 description 3
- 208000025865 Ulcer Diseases 0.000 description 3
- 230000001476 alcoholic effect Effects 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000002296 dynamic light scattering Methods 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 230000008733 trauma Effects 0.000 description 3
- 231100000397 ulcer Toxicity 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 208000004232 Enteritis Diseases 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 229920002752 Konjac Polymers 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 229960003276 erythromycin Drugs 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 230000002550 fecal effect Effects 0.000 description 2
- 230000001279 glycosylating effect Effects 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 239000002674 ointment Substances 0.000 description 2
- 125000002971 oxazolyl group Chemical group 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- TWNIBLMWSKIRAT-RWOPYEJCSA-N (1r,2s,3s,4s,5r)-6,8-dioxabicyclo[3.2.1]octane-2,3,4-triol Chemical compound O1[C@@]2([H])OC[C@]1([H])[C@@H](O)[C@H](O)[C@@H]2O TWNIBLMWSKIRAT-RWOPYEJCSA-N 0.000 description 1
- 235000001206 Amorphophallus rivieri Nutrition 0.000 description 1
- 244000247812 Amorphophallus rivieri Species 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 229910014571 C—O—Si Inorganic materials 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 208000016285 Movement disease Diseases 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 244000170916 Paeonia officinalis Species 0.000 description 1
- 235000006484 Paeonia officinalis Nutrition 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 1
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 241000256856 Vespidae Species 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 208000034158 bleeding Diseases 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 208000037893 chronic inflammatory disorder Diseases 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000000254 damaging effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000000762 glandular Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000033687 granuloma formation Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- NUHSROFQTUXZQQ-UHFFFAOYSA-N isopentenyl diphosphate Chemical compound CC(=C)CCO[P@](O)(=O)OP(O)(O)=O NUHSROFQTUXZQQ-UHFFFAOYSA-N 0.000 description 1
- 239000000252 konjac Substances 0.000 description 1
- 235000010485 konjac Nutrition 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 210000003024 peritoneal macrophage Anatomy 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 239000013585 weight reducing agent Substances 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
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/736—Glucomannans or galactomannans, e.g. locust bean gum, guar gum
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses the glycosylated SiO of Glucomannan2Nano particle and its preparation method and application.A kind of glycosylated SiO of Glucomannan2Nano particle, by SKGM or bletilla polysaccharide and SiO2The particle that obtained particle diameter is 10 1000nm is connected with each other by covalent bond.The invention provides a kind of new silica nanometer polysaccharide, pass through the various sizes of nano SiO 2 particle formation nanometer polysaccharide hydroxy activated with CDI after SKGM containing a large amount of mannoses or bletilla polysaccharide amination, can be with the character mutation of activating macrophage, and then cause new biological effect, improve the inflammatory conditions of intestines and stomach；Simultaneously in wound healing process, by the phenotype for regulating and controlling macrophage, tissue repair type macrophage is formed, is adjusted and acceleration of tissue repair, regeneration, can be applied in terms of the medicine of related inflammation disease and wound healing or external medical apparatus is prepared by TGF β 1 and VEGF.
Description
Technical field
The invention belongs to biomedicine technical field, it is related to the glycosylated SiO of Glucomannan2Nano particle and its preparation side
Method and application.
Background technology
Macrophage (Macrophage) is exempting from of being played an important role in all kinds of chronic inflammatory diseases develop
Epidemic disease cell, including atherosclerosis, asthma, inflammatory gastrointestinal are scorching, rheumatoid arthritis and tissue repair etc..What is activated is huge
Phagocyte stimulates a large amount of secretions of proinflammatory factor, so as to promote inflammatory reaction to develop.When body, inner tissue is inflamed
When, the monocyte in blood is recruited to inflammation part, and is divided into the macrophage of activation, infiltrates mucous layer, and secretion promotees
Inflammatory cytokines, so as to cause epithelial cell and body of gland apoptosis, barrier function forfeiture, necrosis, Granuloma formation and fiber
Change.Such as IBD (Inflammatory bowel disease) and gastric ulcer (Gastric ulcer) are typical slow
Property, the diseases associated with inflammation easily recurred, its cause of disease and pathogenesis are more complicated, and diagnosis and treatment are also quite intractable, but are directed to
The treatment of macrophage is one of current popular research direction, suppresses the activation of macrophage by novel molecular therapeutic strategy
State, reduces the expression of inflammatory factor, so as to alleviate the development of diseases associated with inflammation；Wound healing (wound healing)
It is each stage healed after a complicated biological process, wound to have substantial amounts of protein molecular to be induced and regulated and controled,
These factors are after synthesis and secretion, by acting on body cell and cytoplasm, the balanced development of organization of regulation control healing.Both
Tissue repair is stimulated, the hyperplasia of tissue is limited again, to maintain the dynamic equilibrium of healing.Macrophage by Phenotypic change and
The expression of relevant cell factor, participates in whole tissue repair, the process of regeneration (Tissue repair, Regeneration).
SKGM (Konjac glucomannan, hereinafter referred to as KGM) is neutral nonionic type linear polysaccharide, is by Portugal
Grape sugar and mannose a kind of high-molecular compound with reference to formed by with β-Isosorbide-5-Nitrae glycosidic bond, there is very high viscosity, gelling, hold
Aqueous, biocompatibility etc., these physicochemical properties are with good application prospect.KGM has multiple biological activities, such as anti-
Tumour, improvement function of intestinal canal, fat-reducing, hypoglycemic effect etc., have been applied to clinical, medicine and other fields, to the further of KGM
Research and deep development utilize the extensive concern for having caused people.In recent years, the research of polysaccharide polymer enters one newly
Level, be it in medicine, food with the further investigation to its structure-activity relationship, it was found that the mechanism of action of Pharmacological
The application in product field provides theoretical foundation, accelerates the exploitation of natural polysaccharide polymers.The bioactivity of polysaccharide is relied on
It is mutually distinguishable with acceptor, interact during, and natural poly- polysaccharide greatly limit to itself as macromolecule
The exploration of bioactivity.Bletilla polysaccharide (Bletilla Striata polysaccharide, hereinafter referred to as BSP) be it is a kind of from
Mannosym, the neutral nonionic molded line being made up of 4 molecule mannoses and 1 molecule glucose obtained by the extraction of the traditional Chinese medicine bletilla striata
Shape mannosan, with anti-inflammatory, promotees blood coagulation, antiviral, antitumor, the biological activity such as anti-oxidant.Bletilla polysaccharide is safe height
The medical material of effect, with excellent physicochemical property, the quite biomaterial with development prospect.
Functional molecular is introduced to nano grain surface by covalent bond to obtain as a kind of method for preparing new material
The extensive concern of people and further investigation, and made important progress in fields such as catalysis, electronics and biological and chemical detections,
The particle of this modified can be applied directly as a kind of functional material.SiO2Nano particle is due to its excellent physics
Chemical property, from industrial production to basic research, is all widely used in various fields.Particularly monodispersed SiO2Colloid
Particle, with monodispersity, size is in big scope adjustability, with monodispersed SiO2Nano particle be unit construct it is new
Nano material has potential application prospect.At present, about study limitation of the mannocarolose on macrophage in targeted drug
The structure of carrier.
The content of the invention
It is an object of the invention to provide a kind of glycosylated SiO of Glucomannan2Nano particle.
It is a further object of the present invention to provide the glycosylated SiO of the Glucomannan2The preparation method of nano particle.
It is yet another object of the invention to provide the glycosylated SiO of the Glucomannan2The application of nano particle.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of glycosylated SiO of Glucomannan2Nano particle, by Glucomannan and SiO2It is connected with each other by covalent bond
The particle diameter arrived is 10-1000nm particle.
The glycosylated SiO of described Glucomannan2Nano particle, preferably passes through different SiO2Size Control Glucomannan sugar
The SiO of base2The particle diameter of nano particle.
The glycosylated SiO of described Glucomannan2What nano particle was preferably prepared by the following method arrives：
(1) amination Glucomannan is prepared；
(2) CDI methods activation SiO2Surface hydroxyl obtains the SiO containing imidazole group2；
(3) by amidized Glucomannan and the SiO containing imidazole group2Reaction obtains the glycosylated SiO of Glucomannan2
Nano particle.
The glycosylated SiO of described Glucomannan2What nano particle was further preferably prepared by the following method arrives：
(1) prepared by amination Glucomannan
Glucomannan is weighed in DMSO, 2~4h is dissolved by heating, 40~60 DEG C, is cooled to room temperature, adds excessive carbonyl
Diimidazole (CDI), 2~4h of room temperature reaction is fully activated, and adds excessive ethylenediamine, and continuing stirring makes it fully react, thoroughly
Analysis, is freezed standby；Glucomannan and DMSO mol ratio are 1：106~1：108, the mass ratio of Glucomannan and carbonyl dimidazoles
For 1：2~1：10, Glucomannan is 1 with ethylenediamine mol ratio：105~1：106；
(2) CDI methods activation SiO2Surface hydroxyl
Weigh SiO2Particle is washed for several times, is added anhydrous propanone and is ensured water-less environment, SiO2Mole of particle and anhydrous propanone
Than for 1：40~1:50, add CDI and react 1~2h, anhydrous propanone cleans for several times, removes unnecessary CDI, dry；SiO2With CDI's
Mol ratio is 1：1~1：SiO described in 52The diameter of particle selects 10,30,100,500,1000nm five kinds；
(3)SiO2Surface mannose glycosylation modification
Weigh containing imidazole group SiO2With the DMSO solution containing amination Glucomannan, amination Glucomannan and miaow
Oxazolyl group SiO2Mass ratio be 1：1~1：3, room temperature vortex 2~4h of concussion reaction, that is, obtain the glycosylated SiO of Glucomannan2
Nano particle.
The glycosylated SiO of Glucomannan of the present invention2The preparation method of nano particle is comprised the steps of：
(1) amination Glucomannan is prepared；
(2) CDI methods activation SiO2Surface hydroxyl obtains the SiO containing imidazole group2；
(3) by amidized Glucomannan and the SiO containing imidazole group2Reaction obtains the glycosylated SiO of Glucomannan2
Nano particle.
The glycosylated SiO of Glucomannan of the present invention2The preparation method of nano particle is further preferably comprising following
Step：
(1) prepared by amination Glucomannan
Glucomannan is weighed in DMSO, 2~4h is dissolved by heating, 40~60 DEG C, is cooled to room temperature, adds excessive carbonyl
Diimidazole (CDI), 2~4h of room temperature reaction is fully activated, and adds excessive ethylenediamine, and continuing stirring makes it fully react, thoroughly
Analysis, is freezed standby；Glucomannan and DMSO mol ratio are 1：106~1：108, the mass ratio of Glucomannan and carbonyl dimidazoles
For 1：2~1：10, Glucomannan is 1 with ethylenediamine mol ratio：105~1：106；
(2) CDI methods activation SiO2Surface hydroxyl
Weigh SiO2Particle is washed for several times, is added anhydrous propanone and is ensured water-less environment, SiO2Mole of particle and anhydrous propanone
Than for 1：40~1:50, add CDI and react 1~2h, anhydrous propanone cleans for several times, removes unnecessary CDI, dry；SiO2With CDI's
Mol ratio is 1：1~1：5；Described SiO2The diameter of particle selects 10,30,100,500,1000nm five kinds；
(3)SiO2Surface mannose glycosylation modification
Weigh containing imidazole group SiO2With the DMSO solution containing amination Glucomannan, amination Glucomannan and miaow
Oxazolyl group SiO2Mass ratio be 1：1~1：3, room temperature vortex 2~4h of concussion reaction, that is, obtain the glycosylated SiO of Glucomannan2
Nano particle.
Supernatant further preferably is abandoned comprising centrifugation after 2~4h of room temperature vortex concussion reaction in step (3), then it is repeatedly clear with deionization
Wash, centrifugation is until without unnecessary solvent, reagent, 37 ° of drying are preserved in vacuum drying.
Glucomannan of the present invention glycosylates SiO2Nano particle is preparing treatment diseases associated with inflammation, gastric ulcer, wound
Application in the medicine that recovery is closed.
Described diseases associated with inflammation preferably comprises such as colitis IBD, chronic gastritis, arthritis.
This invention to IBD disease, gastric ulcer model and full thickness skin excision trauma model by carrying out example
Card.Animal herein includes but is not limited to：Mouse, rat, performing animal includes but is not limited to cat, dog, and other one
A little animals for example but are not limited to ox, sheep, pig, horse, and primate for example but is not limited to monkey and people.Mouse inflammatory bowel
Sick disease model, vivo detection be widely recognized as and received internal pharmaceutical activity detection model, while can also be
It is other biological for example but to be not limited to people reference is provided.
Beneficial effect
The invention provides a kind of new silica nanometer polysaccharide, SKGM or the bletilla striata containing a large amount of mannoses are more
By the various sizes of nano SiO 2 particle formation nanometer polysaccharide hydroxy activated with CDI after sugared amination, it can activate
The character mutation of macrophage, and then cause new biological effect, a large amount of suppressive genes IL-10 are such as secreted, improve stomach and intestine
The inflammatory conditions in road；Simultaneously in wound healing process, by regulating and controlling the phenotype of macrophage, tissue repair type macrophage is formed thin
Born of the same parents, are adjusted and acceleration of tissue repair, regeneration by TGF-β 1 and VEGF, can prepare related inflammation disease and wound healing
Applied in terms of the medicine or external medical apparatus of conjunction.The present invention lays the foundation for the bioactivity of further research Glucomannan；
Compared with small-molecule drug, with high raw material selectivity, cost is relatively low, and nanosizing product is single, and separation is simple, possesses one
Fixed industrial prospect；With holosaccharide ratio, what is protruded in therapeutic effect is to have more targeting, and drug effect is more preferably.
Brief description of the drawings
Fig. 1 is the SiO of different-grain diameter2And the KSiNPs synthesized as carrier transmission electron microscope morphology analysis
Fig. 2 is the grain size distribution that dynamic light scattering detects various sizes of SiNPs and KSiNPs
Fig. 3 is that infrared spectrum analysis detects that KSiNP30 specificity absorbs group
Fig. 4 is the efficacy assessment picture that KSiNP30 treats mouse TNBS diseases associated with inflammation
Fig. 5 is the treatment picture that KSiNP30 cuts off trauma model to mouse full thickness skin
Fig. 6 is the curve map that KSiNP30 cuts off wound healing rate to mouse full thickness skin
Fig. 7 is the curve map that BSiNP30 cuts off wound healing rate to mouse full thickness skin
Fig. 8 is influence of the various sizes of nanometer polysaccharide to IL-10 levels in Macrophage Cell supernatant
Embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described with reference to embodiment, still
It should be appreciated that these descriptions are simply to further illustrate the features and advantages of the present invention, rather than to the claims in the present invention
Limitation.
SKGM used in following examples comes from Megazyme (Wicklow, Ireland), and molecular weight is
200kDa, but it is not construed as the limitation to SKGM using source.
Embodiment 1：The preparation of KSiNPs solution
(1) prepared by amination SKGM
A certain amount of SKGM is weighed in DMSO, 3h is dissolved by heating, 45 DEG C, is cooled to room temperature, adds excessive carbonyl
Base diimidazole (CDI), room temperature reaction 3h is fully activated, and adds excessive ethylenediamine, is continued to stir 18h, it is fully reacted,
Dialysis three days, is freezed standby；SKGM and DMSO mol ratio are 5：106, the mass ratio of SKGM and carbonyl dimidazoles
For 1：2, SKGM is 1 with ethylenediamine mol ratio：105。
(2) CDI methods activation SiO2Surface hydroxyl
A certain amount of SiO is weighed respectively2(particle diameter is respectively 10,30,100,500,1000nm) washs for several times, adds certain
The anhydrous propanone of amount ensures water-less environment, adds excess CDI and reacts 1~2h, anhydrous propanone cleans for several times, removes unnecessary CDI, dry in the air
It is dry；SiO2Mol ratio with CDI is 2：1.
(3)SiO2Surface mannose glycosylation modification
Weigh a certain amount of containing imidazole group SiO2With the DMSO solution containing amination SKGM, amination konjaku
Polysaccharide and imidazole group SiO2Mass ratio be 1：2, room temperature vortex concussion reaction 4h, that is, obtain glycosylating SiO2SKGM
Nano particle (hereinafter referred to as KSiNPs).
The physicochemical property and sign of various sizes of SKGM nano particle, are shown in Fig. 1, Fig. 2 and Fig. 3
Transmission electron microscope results such as Fig. 1, the polysaccharide nanoparticle size of synthesis is substantially without significant change, surface attachment polysaccharide,
Particle diameter distribution also no significant difference.
The particle diameter distribution of different size silicas before and after dynamic light scattering detection modification, as a result such as Fig. 2, receiving after modification
Rice grain size is distributed also without significant change without significant change.
The result of infrared spectrogram such as Fig. 3, SiO2In 3 445.4cm-1Locate appearance-OH characteristic absorption peak, it can thus be appreciated that
SiO2Nano-particle surface is present after substantial amounts of hydroxyl, modified polysaccharide in 3 100~3 450cm-1The hydroxyl peak at place substantially dies down；
SiO2In 1094.7cm-1There is a wide strong asymmetric absworption peak, this is by Si-O-Si symmetrical stretching vibration absworption peaks.
462.6cm-1Locate the flexural vibrations absworption peak for Si-O-Si.And in Silica-KGM infrared spectrum, except original characteristic peak
Outside, in 2876.1cm-1Occur in that very strong-C-H ,-CH2- stretching vibration absworption peak, and in 1400cm-1There is C-H in left and right
Inplane shear vibration absorption peak.In addition in 1097.7cm-1, -1065.8cm-1It is interior multiple strong absworption peak occur, this be C-O-C,
Si-O-C and Si-O-Si symmetrical stretching vibration absworption peak, these illustrate SiO2Reacted with KGM ,-(OCH2CH2)n-
Strand has been integrated into SiO2On, KGM is coated on Nano-meter SiO_22On surface.This result shows the SiO of imidazole radicals activation2With
Hydroxyl on amination SKGM is reacted, and has synthesized the nanometer polysaccharide that we want.
Embodiment 2：The preparation of BSiNPs solution
(1) prepared by amination bletilla polysaccharide
A certain amount of bletilla polysaccharide is weighed in DMSO, 3h is dissolved by heating, 45 DEG C, is cooled to room temperature, adds excessive carbonyl
Base diimidazole (CDI), room temperature reaction 3h is fully activated, and adds excessive ethylenediamine, is continued to stir 18h, it is fully reacted,
Dialysis three days, is freezed standby；Bletilla polysaccharide and DMSO mol ratio are 5：106, the mass ratio of bletilla polysaccharide and carbonyl dimidazoles
For 1：2, bletilla polysaccharide is 1 with ethylenediamine mol ratio：105。
(2) CDI methods activation SiO2Surface hydroxyl
Particle diameter is weighed for 30nmSiO2Washing for several times, adds a certain amount of anhydrous propanone and ensures water-less environment, adds excessive
CDI reacts 2h, and anhydrous propanone cleans for several times, removes unnecessary CDI, dry；SiO2Mol ratio with CDI is 2：1.
(3)SiO2Surface mannose glycosylation modification
Weigh a certain amount of containing imidazole group SiO2With the DMSO solution containing amination bletilla polysaccharide, the amination bletilla striata
Polysaccharide and imidazole group SiO2Mass ratio be 1：3, room temperature vortex concussion reaction 4h, that is, obtain glycosylating SiO2Bletilla polysaccharide
Nano particle (hereinafter referred to as BSiNPs).
The particle diameter distribution of different size silicas before and after modification is detected by dynamic light scattering, is as a result shown after modification
Nanoparticle size is distributed also without significant change without significant change.
Embodiment 3：Therapeutic actions of the KSiNPs/BSiNPs to mouse TNBS enteritis
Mouse TNBS colitis models are set up according to literature procedure, that is, take female BAl BIc/C mice 50, body weight 16-
18g, is randomly divided into 5 groups, fasting 24 hours, bowel lavage administration TNBS solution sets up TNBS colitis models by mouse.
12 hours before and after TNBS bowel lavage of the BALB/c mouse through 3mg/ml, bowel lavage administration, is grouped as follows respectively：
Blank control group：Poured into when model is set up 50% ethanol solution, bowel lavage be administered when feed physiological saline；
Model group：3mg/ml TNBS is poured into when model is set up, bowel lavage feeds physiological saline when being administered；
SKGM group：3mg/ml TNBS is poured into when model is set up, bowel lavage feeds KGM solution as control when being administered；
Bletilla polysaccharide group：3mg/ml TNBS is poured into when model is set up, bowel lavage feeds BSP solution as control when being administered；
SiO2Group：3mg/ml TNBS is poured into when model is set up, bowel lavage feeds the SiO of Isodose when being administered2Solution conduct
Control；
KSiNPs groups：3mg/ml TNBS is poured into when model is set up, bowel lavage feeds KSiNP30 when being administered, and (embodiment 1 is made
Standby, particle diameter is 30nm) (50mg/kg)
BSiNPs groups：3mg/ml TNBS is poured into when model is set up, bowel lavage feeds BSiNP30 when being administered, and (embodiment 2 is made
Standby, particle diameter is 30nm) (50mg/kg)
It is three administrations in TNBS models, administered volume is 100 μ l.Mouse hair situation is observed after administration daily,
Fecal character, weighs mouse weight, and record in detail, three days altogether.DAI standards of grading such as following table.If observing mouse weight,
Mouse feeder is set up the 8th day to model.
(1) disease activity (DAI) scores
Observation mouse hair situation, fecal character, weighing mouse weight, and record in detail, three days altogether daily after administration.
DAI standards of grading such as following table.
The DAI of table 1 integrates point-score
DAI scoring=(weight loss integration+excrement continuous integral+occult blood naked eyes blood integration)/3
(2) colon HE pathological sections score
Put to death animal within 3 days after model is set up, take mouse Colon, 4% formaldehyde fixes 12 hours, embedded section, H＆E dyeing,
Its pathological change of micro- sem observation, and scored.Standards of grading are as follows：
The mouse Colon HE pathological section standards of grading of table 2：
After TNBS bowel lavage modelings, there is obvious diarrhoea in mouse, body weight reduction, and the symptom such as have blood in stool, DAI scorings can be integrated
State the disease severity of metrics evaluation mouse colitis.As shown in table 3, the DAI scorings of TNBS model mices and normal health
Mouse contrast is notable to be risen, and SKGM nano particle (KSiNP30) and bletilla polysaccharide nano particle (BSiNP30) treatment can
Effectively to alleviate enteritis symptom, reduction DAI scores, in addition SKGM, bletilla polysaccharide and SiO2Processing is to the no shadow of DAI scorings
Ring.By histological score, we can be found that rarely seen a small amount of inflammatory cell infiltration in treatment group, and histological score is substantially less than
TNBS model groups；And SKGM, bletilla polysaccharide and SiO2Individually processing has no influence to the histological scores of colon.Thus may be used
See that KSiNPs and BSiNPs treatments can effectively alleviate the intestinal inflammatory of TNBS inductions.
The SKGM nano particle of table 3 treats TNBS colitis model mouse curative effects
Data are shown that significant difference is examined by ANOVA and determined in the form of mean+SD.
Compared with model group, * represents P≤0.05
KSiNPs group treatment results are taken pictures such as Fig. 4, and control mice colon lengths are longer, without red and swollen phenomenon, without substantially other
Inflammatory symptom.TNBS groups inflammatory symptom substantially, there is redness, the phenomenon that colon shortens.The inflammatory symptom of KGM groups is present, but does not have
TNBS group inflammation is obvious, it may be possible to because KGM plays the role of to improve intestinal environment.Glycosylate SiO2SKGM nanometer
Grain group (SiO2- KGM groups) to it is normal similar, without substantially red and swollen phenomenon.SiO is glycosylated as can be seen here2SKGM nano particle
Truly have the effect for alleviating inflammation.
Embodiment 4：Therapeutic actions of the KSiNP30 to mouse Alcoholic gastric ulcer model
Mouse Alcoholic gastric ulcer model is set up according to literature procedure, that is, takes ICR male mices 50, body weight 20-
22g, 5 groups are randomly divided into by mouse, based on Alcoholic gastric ulcer model gastric infusion.
ICR mouse were through fasting in 24 hours, and gastric infusion, is grouped as follows respectively：,
Blank control group：Physiological saline is fed during gastric infusion；
Model group：Physiological saline is fed during gastric infusion；
SKGM group：KGM solution is fed during gastric infusion as control；
Bletilla polysaccharide group：BSP solution is fed during gastric infusion as control；；
SiO2Group：The SiO of Isodose is fed during gastric infusion2Solution is used as control；
KSiNPs groups：Gastric infusion feeds KSiNP30 (prepared by embodiment 1, particle diameter is 30nm) 50mg/kg.
BSiNPs groups：Gastric infusion feeds BSiNP30 (prepared by embodiment 2, particle diameter is 30nm) 50mg/kg.
After administration 4 hours, in addition to blank control group, 0.2ml/ absolute ethyl alcohol gavage of remaining each group observes mouse state
And record, then put to death mouse；Orifice of the stomach is ligatured after execution, full stomach is taken out, fixes 1 hour, cleaned, visually observed with physiological saline
Gastric ulcer situation, records ulcer index.Model group half mouse occur prostrate, tremble, movement disorder symptom；Body of stomach, which goes out, larger
There is petechial hemorrhage in petechial hemorrhage or bleeding band, small part, it is seen that glandular stomach portion mucous membrane peony, there is substantially fair mass formed by blood stasis shape.
KSiNPs has good protective effect under dosage used to mouse absolute ethyl alcohol model, and compared with blank control group has to ulcer
Preferable inhibiting rate, can mitigate damaging action of the ethanol to stomach lining, and ulcer index is significantly lower than control group.It the results are shown in Table 4.
Influence of the SKGM nano particle of table 4 to mouse alcohol gastric ulcer
Data are shown that significant difference is examined by ANOVA and determined in the form of mean+SD.
Compared with model group, * represents P≤0.05
Embodiment 5：KSiNP30 cuts off the therapeutic action of trauma model to mouse full thickness skin
Mouse skin wound healing is set up according to literature procedure：
A. C57 mouse are taken, yellow Jackets intraperitoneal anesthesia, back unhairing, sterilization makes a call to one with special card punch in mouse back
Circular hole, cuts diameter 7mm, area average out to mm2Skin, be deep to subcutaneous, after hemostasis, take pictures and be designated as Day1, control group is with one
Secondary property aseptic dressing patch wrapping, single cage is fed.
B. medicine is applied to wound by experimental group, then is wrapped up with the gauze for coating erythromycin ointment, and single cage is fed.
C. take a picture within 3 days, 5 days, 7 days, 9 days and 12 days after wound, then using the R Plus of U.S. Image-Pro zero
Version6 (IPP) image analysis software measures the wound area of mouse sum.Healing rate=[(original surface of a wound area-do not heal wound
Face area) the original surface of a wound areas of ÷] × 100%.
Take wound tissue, fixed, specimens paraffin embedding slices, HE dyeing and sirius red dyeing, observe the surface of a wound pathology and
Cytologic characteristic.
Experiment is randomly divided into four groups：
Control group：Physiological saline is fed when smearing administration；
SKGM group：KGM solution is fed as control when smearing administration；
Bletilla polysaccharide group：BSP solution is fed as control when smearing administration；
SiO2Group：The SiO of Isodose is fed when smearing administration2Solution is used as control；
KSiNPs groups：KSiNPs (prepared by embodiment 1, particle diameter is 30nm) 5mg/ml is fed when smearing administration；
BSiNPs groups：BSiNPs (prepared by embodiment 1, particle diameter is 30nm) 5mg/ml is fed when smearing administration.
Smear after administration, erythromycin ointment smears wound, disposable sterilized dressing patch wrapping, single cage is raised, and observes mouse
The change of the forward and backward body weight of modeling, fur, motion and feed.Pass through Wound healing rate, wound healing time and histopathology credit
Analysis is used as directly effective Evaluation Criterion of Wound Healing.The result of wound healing is shown in Fig. 5, it is seen that pair of the healing rate of wound
Than obvious, the wound healing speed of KSiNP groups is substantially accelerated compared with other several groups, illustrates that the nano particles of many saccharification can speed up wound
Mouth healing.
The wound healing rate of the statistics of wound healing rate such as Fig. 6 and 7, KSiNPs group and BSiNPs groups has bright in wound early stage
The aobvious effect for improving healing, illustrates that nanometer polysaccharide has the effect of promotion in wound healing.
The cytokine assay of embodiment 6
IL-10 is important suppression inflammatory factor, can alleviate inflammatory conditions, embodies the biology effect of nanometer polysaccharide
Should.
Peritoneal macrophage is carried, is spread in 48 orifice plates, cell concentration is more than 80%, plus different sizes prepared by embodiment 1
KSiNPs nano particles (100ug/ml) stimulate 24 hours after receive sample, and set Control groups, ELISA kit detection IL-
The inflammation-related factors such as 10, TNF-a.As a result Fig. 8 is seen, KSiNP30 is than control and the KSiNPs nano particle energy of other particle diameters
Notable stimulating expression of macrophage secretes a large amount of suppressive genes IL-10.
BSiNP30, KSiNP30 and control SiO in above example 3-62Particle is by first using ethanol：Dd water=4：
1 cleaning three times, then with normal saline into certain suitable concn.By KSiNP30 (prepared by embodiment 1, particle diameter is 30nm) or
SiO2Particle is dispersed in PBS solution, ultrasonic disperse before administration.The KGM of SKGM group and the BSP of bletilla polysaccharide group are then direct
With preparing solutions for administration after ultraviolet irradiation.
In subsequent research, inventor has also investigated 10,100,500,1000nm KSiNPs and BSiNPs to colon
Scorching, gastric ulcer and the curative effect of wound healing, have obtained the result being close with KSiNP30 and BSiNP30, certain KSiNPs and
BSiNPs is significantly superior to KGM and BSP controls to colitis, gastric ulcer and the curative effect of wound healing, also has and be significantly superior to
SiO2Control.
Claims (9)
1. a kind of glycosylated SiO of Glucomannan2Nano particle, it is characterised in that by Glucomannan and SiO2Pass through covalent bond phase
The particle that the obtained particle diameter that connects is 10-1000nm；Described Glucomannan is selected from SKGM or bletilla polysaccharide.
2. the glycosylated SiO of Glucomannan according to claim 12Nano particle, it is characterised in that pass through different SiO2Chi
The glycosylated SiO of very little control Glucomannan2The particle diameter of nano particle.
3. the glycosylated SiO of Glucomannan according to claim 12Nano particle, it is characterised in that make by the following method
It is standby to obtain：
(1) amination Glucomannan is prepared；
(2) CDI methods activation SiO2Surface hydroxyl obtains the SiO containing imidazole group2；
(3) by amidized Glucomannan and the SiO containing imidazole group2Reaction obtains the glycosylated SiO of Glucomannan2Nanometer
Particle.
4. the glycosylated SiO of Glucomannan according to claim 32Nano particle, it is characterised in that make by the following method
It is standby to obtain：
(1) prepared by amination Glucomannan：
Glucomannan is weighed in DMSO, 2~4h is dissolved by heating, 40~60 DEG C, is cooled to room temperature, adds the excessive miaow of carbonyl two
Azoles CDI, 2~4h of room temperature reaction is fully activated, and adds excessive ethylenediamine, and continuing stirring makes it fully react, and dialyses, and freezes
It is standby；Glucomannan and DMSO mol ratio are 1：106~1：108, the mass ratio of Glucomannan and carbonyl dimidazoles is 1：2~
1：10, Glucomannan is 1 with ethylenediamine mol ratio：105~1：106；
(2) CDI methods activation SiO2Surface hydroxyl
Weigh SiO2Particle is washed for several times, is added anhydrous propanone and is ensured water-less environment, SiO2The mol ratio of particle and anhydrous propanone is
1：40~1:50, add CDI and react 1~2h, anhydrous propanone cleans for several times, removes unnecessary CDI, dry；SiO2With CDI mole
Than for 1：1~1：5；Described SiO2The diameter of particle is selected from 10,30,100,500 or 1000nm；
(3)SiO2Surface mannose glycosylation modification
Weigh containing imidazole group SiO2With the DMSO solution containing amination Glucomannan, amination Glucomannan and imidazole radicals
Group SiO2Mass ratio be 1：1~1：3, room temperature vortex 2~4h of concussion reaction, that is, obtain the glycosylated SiO of Glucomannan2Nanometer
Particle.
5. the glycosylated SiO of Glucomannan described in claim 12The preparation method of nano particle, it is characterised in that comprising following
Step：
(1) amination Glucomannan is prepared；
(2) CDI methods activation SiO2Surface hydroxyl obtains the SiO containing imidazole group2；
(3) by amidized Glucomannan and the SiO containing imidazole group2Reaction obtains the glycosylated SiO of Glucomannan2Nanometer
Particle；
Described Glucomannan is selected from SKGM or bletilla polysaccharide.
6. preparation method according to claim 5, it is characterised in that be prepared by the following method and obtain：
(1) prepared by amination Glucomannan：
Glucomannan is weighed in DMSO, 2~4h is dissolved by heating, 40~60 DEG C, is cooled to room temperature, adds the excessive miaow of carbonyl two
Azoles CDI, 2~4h of room temperature reaction is fully activated, and adds excessive ethylenediamine, and continuing stirring makes it fully react, and dialyses, and freezes
It is standby；Glucomannan and DMSO mol ratio are 1：106~1：108, the mass ratio of Glucomannan and carbonyl dimidazoles is 1：2~
1：4, Glucomannan is 1 with ethylenediamine mol ratio：105~1：106；
(2) CDI methods activation SiO2Surface hydroxyl
Weigh SiO2Particle is washed for several times, is added anhydrous propanone and is ensured water-less environment, SiO2The mol ratio of particle and anhydrous propanone is
1：40~1:50, add CDI and react 1~2h, anhydrous propanone cleans for several times, removes unnecessary CDI, dry；SiO2With CDI mole
Than for 2：1~1：1；Described SiO2The diameter of particle is selected from 10,30,100,500 or 1000nm；
(3)SiO2Surface mannose glycosylation modification
Weigh containing imidazole group SiO2With the DMSO solution containing amination Glucomannan, amination Glucomannan and imidazole radicals
Group SiO2Mass ratio be 1：1~1：3, room temperature vortex 2~4h of concussion reaction, that is, obtain the glycosylated SiO of Glucomannan2Nanometer
Particle.
7. preparation method according to claim 6, it is characterised in that in step (3) after 2~4h of room temperature vortex concussion reaction
Supernatant also is abandoned comprising centrifugation, then is cleaned repeatedly with deionization, centrifugation is until without unnecessary solvent, reagent, 37 DEG C of drying, in vacuum
The step of kept dry.
8. the glycosylated SiO of Glucomannan described in claim 12Nano particle prepare treatment diseases associated with inflammation, gastric ulcer,
Application in the medicine and/or medical dressing of wound healing.
9. application according to claim 8, it is characterised in that described diseases associated with inflammation includes：Colitis, chronic gastritis,
Arthritis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710424892.1A CN107158406B (en) | 2017-06-06 | 2017-06-06 | Glucomannan glycosylated SiO2 nano-particle and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710424892.1A CN107158406B (en) | 2017-06-06 | 2017-06-06 | Glucomannan glycosylated SiO2 nano-particle and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107158406A true CN107158406A (en) | 2017-09-15 |
| CN107158406B CN107158406B (en) | 2020-07-14 |
Family
ID=59825408
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710424892.1A Active CN107158406B (en) | 2017-06-06 | 2017-06-06 | Glucomannan glycosylated SiO2 nano-particle and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107158406B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276553A (en) * | 2020-12-31 | 2022-04-05 | 南京工业大学 | Monosaccharide functionalized organic nano soft ball and preparation method thereof |
| CN116333575A (en) * | 2023-03-27 | 2023-06-27 | 上海云谈科技有限公司 | Green multi-substrate universal rainproof and stain-resistant light-transmitting coating for solar power supply wireless communication system and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709915A (en) * | 2005-06-23 | 2005-12-21 | 南京大学医学院附属鼓楼医院 | Polysaccharide material for promoting cell tactophily and preparation thereof |
| CN101002793A (en) * | 2007-01-10 | 2007-07-25 | 南京大学 | Application of polysaccharide of bletilla striata for preparing medicines to treat peptic ulcer |
| CN101249274A (en) * | 2008-04-01 | 2008-08-27 | 南京大学 | Preparation of bletilla striata polyose water gelatin of promoting wound healing and uses thereof |
-
2017
- 2017-06-06 CN CN201710424892.1A patent/CN107158406B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709915A (en) * | 2005-06-23 | 2005-12-21 | 南京大学医学院附属鼓楼医院 | Polysaccharide material for promoting cell tactophily and preparation thereof |
| CN101002793A (en) * | 2007-01-10 | 2007-07-25 | 南京大学 | Application of polysaccharide of bletilla striata for preparing medicines to treat peptic ulcer |
| CN101249274A (en) * | 2008-04-01 | 2008-08-27 | 南京大学 | Preparation of bletilla striata polyose water gelatin of promoting wound healing and uses thereof |
Non-Patent Citations (2)
| Title |
|---|
| CHAO WANG ET AL.: ""Preparation and Characterization of Konjac Glucomannan (KGM)/Sio2 Nanopaticles Blend Film"", 《ADVANCED MATERIALS RESEARCH》 * |
| ZHENZHEN WANG ET AL.: ""Specifically Formed Corona on Silica Nanoparticles Enhances Transforming Growth Factor β1 Activity in Triggering Lung Fibrosis"", 《ACS NANO》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276553A (en) * | 2020-12-31 | 2022-04-05 | 南京工业大学 | Monosaccharide functionalized organic nano soft ball and preparation method thereof |
| CN114276553B (en) * | 2020-12-31 | 2023-02-21 | 南京工业大学 | Monosaccharide functionalized organic nano soft ball and preparation method thereof |
| CN116333575A (en) * | 2023-03-27 | 2023-06-27 | 上海云谈科技有限公司 | Green multi-substrate universal rainproof and stain-resistant light-transmitting coating for solar power supply wireless communication system and preparation method thereof |
| CN116333575B (en) * | 2023-03-27 | 2024-03-26 | 上海云谈科技有限公司 | Green multi-substrate universal rainproof and stain-resistant light-transmitting coating for solar power supply wireless communication system and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107158406B (en) | 2020-07-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Chitosan: Structural modification, biological activity and application | |
| Hu et al. | Formation of self-assembled polyelectrolyte complex hydrogel derived from salecan and chitosan for sustained release of Vitamin C | |
| Jiang et al. | Effect of chitosan oligosaccharide-conjugated selenium on improving immune function and blocking gastric cancer growth | |
| Ullah et al. | Natural and synthetic materials based cmch/PVA hydrogels for oxaliplatin delivery: Fabrication, characterization, in-vitro and in-vivo safety profiling | |
| Kesavan et al. | Ulvan loaded graphene oxide nanoparticle fabricated with chitosan and d-mannose for targeted anticancer drug delivery | |
| CN105534957B (en) | A kind of core-shell structure nanometer particle of reduction/enzyme/pH multiple responses drug release | |
| Ye et al. | Phosphorylation and anti-tumor activity of exopolysaccharide from Lachnum YM120 | |
| CN105816920A (en) | Preparation method of modified sodium alginate embolization microspheres | |
| JP2008260823A (en) | Polysaccharide modified with selenious acid and its manufacturing method | |
| EP1455802A1 (en) | Chitosan oligosaccharides and uses thereof | |
| CN112516323B (en) | Preparation method of photo-crosslinked chitosan-methacrylic acid nanoparticle | |
| CN108178803A (en) | A kind of preparation of cinnamic acid-dextran polymer self-assembled nanometer grain for carrying medicine and its antitumor application thereof | |
| CN107158406A (en) | The glycosylated SiO of Glucomannan2Nano particle and its preparation method and application | |
| JP4321007B2 (en) | Polysaccharide complex and method for producing the same | |
| CN101658484B (en) | Preparation method and application of chitosan copolymer carrier micelle modified by cholesterol formyl chloride | |
| Ostovar et al. | Nanocomposite of chitosan/gelatin/carbon quantum dots as a biocompatible and efficient nanocarrier for improving the Curcumin delivery restrictions to treat brain cancer | |
| CN104434792B (en) | Polymer micelle and preparation method thereof and antineoplastic pharmaceutical compositions, preparation and preparation method thereof | |
| Singh et al. | Recent progress on modified gum katira polysaccharides and their various potential applications | |
| Yuan et al. | Evaluation and mechanism of immune enhancement effects of Pleurotus ferulae polysaccharides-gold nanoparticles | |
| Verma et al. | 1, 3-Beta-Glucans: Drug Delivery and Pharmacology | |
| CN105860103A (en) | Preparation method and application of novel nano hydrogel | |
| Singha et al. | Applications of alginate-based bionanocomposites in drug delivery | |
| CN105056239B (en) | The composite material and its preparation of functional mesoporous silicon dioxide carried drug and siRNA and the application in preparing anticancer drug | |
| CN115252797A (en) | Novel multifunctional nano-drug carrier, preparation method and application | |
| CN107007555A (en) | It is acylated the preparation and application of Glucomannan nano particle |
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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20180223 Address after: No. 8, Ningguo City Economic and Technological Development Zone, Xuancheng, Anhui Province Applicant after: NINGGUO WUYUE MEDICAL TECHNOLOGY CO.,LTD. Address before: Qixia Xianlin Avenue District of Nanjing City, Jiangsu Province, Nanjing University No. 163 210023 Applicant before: Nanjing University |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231101 Address after: 230601 First Floor, Building 2-A, Furong Road North, Economic and Technological Development Zone, Hefei City, Anhui Province Patentee after: HEFEI DAMI MEDICAL SCIENCE & TECHNOLOGY Co.,Ltd. Address before: No. 8 Chuangye Road, Ningguo Economic and Technological Development Zone, Xuancheng City, Anhui Province, 242300 Patentee before: NINGGUO WUYUE MEDICAL TECHNOLOGY CO.,LTD. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231222 Address after: No. 88 Dong'an Road, Anzhen Street, Xishan District, Wuxi City, Jiangsu Province, 214104 Patentee after: Wuxi Kanghe Qingyuan Biotechnology Co.,Ltd. Address before: 230601 First Floor, Building 2-A, Furong Road North, Economic and Technological Development Zone, Hefei City, Anhui Province Patentee before: HEFEI DAMI MEDICAL SCIENCE & TECHNOLOGY Co.,Ltd. |
|
| TR01 | Transfer of patent right |