CN114224911A - Nano material capable of efficiently removing cfDNA and ROS, and preparation method and application thereof - Google Patents
Nano material capable of efficiently removing cfDNA and ROS, and preparation method and application thereof Download PDFInfo
- Publication number
- CN114224911A CN114224911A CN202111425453.5A CN202111425453A CN114224911A CN 114224911 A CN114224911 A CN 114224911A CN 202111425453 A CN202111425453 A CN 202111425453A CN 114224911 A CN114224911 A CN 114224911A
- Authority
- CN
- China
- Prior art keywords
- ros
- cfdna
- nano material
- bowel disease
- msn
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000002086 nanomaterial Substances 0.000 title claims abstract description 128
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 238000011282 treatment Methods 0.000 claims abstract description 60
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims abstract description 58
- BTJZPBCHIAALMP-UHFFFAOYSA-N C(C)O[Si](CCC[Se][Se]CCC[Si](OCC)(OCC)OCC)(OCC)OCC Chemical compound C(C)O[Si](CCC[Se][Se]CCC[Si](OCC)(OCC)OCC)(OCC)OCC BTJZPBCHIAALMP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000002260 anti-inflammatory agent Substances 0.000 claims abstract description 12
- 229940124599 anti-inflammatory drug Drugs 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims description 44
- 229920002873 Polyethylenimine Polymers 0.000 claims description 31
- 229940079593 drug Drugs 0.000 claims description 25
- KBOPZPXVLCULAV-UHFFFAOYSA-N mesalamine Chemical compound NC1=CC=C(O)C(C(O)=O)=C1 KBOPZPXVLCULAV-UHFFFAOYSA-N 0.000 claims description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 229960004963 mesalazine Drugs 0.000 claims description 21
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 18
- 208000012876 acute enteritis Diseases 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 18
- 239000000243 solution Substances 0.000 claims description 15
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 13
- 239000012043 crude product Substances 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 12
- 230000001154 acute effect Effects 0.000 claims description 11
- 238000010992 reflux Methods 0.000 claims description 11
- 230000002000 scavenging effect Effects 0.000 claims description 11
- 150000003384 small molecules Chemical class 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 9
- 230000001684 chronic effect Effects 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 230000002265 prevention Effects 0.000 claims description 8
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 229910001868 water Inorganic materials 0.000 claims description 7
- MQUNPMBEKMVOHA-UHFFFAOYSA-N (sodiodiselanyl)sodium Chemical compound [Na][Se][Se][Na] MQUNPMBEKMVOHA-UHFFFAOYSA-N 0.000 claims description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 5
- 239000002105 nanoparticle Substances 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- OXYZDRAJMHGSMW-UHFFFAOYSA-N 3-chloropropyl(trimethoxy)silane Chemical compound CO[Si](OC)(OC)CCCCl OXYZDRAJMHGSMW-UHFFFAOYSA-N 0.000 claims description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 4
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 4
- DDEHZCWFAUZBEH-UHFFFAOYSA-O azanium;ethanol;nitrate Chemical compound [NH4+].CCO.[O-][N+]([O-])=O DDEHZCWFAUZBEH-UHFFFAOYSA-O 0.000 claims description 4
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000004593 Epoxy Substances 0.000 claims description 3
- 150000001768 cations Chemical class 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- 229920000962 poly(amidoamine) Polymers 0.000 claims description 3
- UETNIIAIRMUTSM-UHFFFAOYSA-N Jacareubin Natural products CC1(C)OC2=CC3Oc4c(O)c(O)ccc4C(=O)C3C(=C2C=C1)O UETNIIAIRMUTSM-UHFFFAOYSA-N 0.000 claims description 2
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims description 2
- LMEKQMALGUDUQG-UHFFFAOYSA-N azathioprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC=NC2=C1NC=N2 LMEKQMALGUDUQG-UHFFFAOYSA-N 0.000 claims description 2
- 229960002170 azathioprine Drugs 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- 229960000485 methotrexate Drugs 0.000 claims description 2
- 229960001940 sulfasalazine Drugs 0.000 claims description 2
- NCEXYHBECQHGNR-QZQOTICOSA-N sulfasalazine Chemical compound C1=C(O)C(C(=O)O)=CC(\N=N\C=2C=CC(=CC=2)S(=O)(=O)NC=2N=CC=CC=2)=C1 NCEXYHBECQHGNR-QZQOTICOSA-N 0.000 claims description 2
- NCEXYHBECQHGNR-UHFFFAOYSA-N sulfasalazine Natural products C1=C(O)C(C(=O)O)=CC(N=NC=2C=CC(=CC=2)S(=O)(=O)NC=2N=CC=CC=2)=C1 NCEXYHBECQHGNR-UHFFFAOYSA-N 0.000 claims description 2
- YUYCVXFAYWRXLS-UHFFFAOYSA-N trimethoxysilane Chemical compound CO[SiH](OC)OC YUYCVXFAYWRXLS-UHFFFAOYSA-N 0.000 claims description 2
- 229940121363 anti-inflammatory agent Drugs 0.000 claims 3
- 239000000825 pharmaceutical preparation Substances 0.000 abstract description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 120
- 241000699670 Mus sp. Species 0.000 description 51
- BLFLLBZGZJTVJG-UHFFFAOYSA-N benzocaine Chemical compound CCOC(=O)C1=CC=C(N)C=C1 BLFLLBZGZJTVJG-UHFFFAOYSA-N 0.000 description 44
- 210000001072 colon Anatomy 0.000 description 44
- 229920003045 dextran sodium sulfate Polymers 0.000 description 44
- 230000000694 effects Effects 0.000 description 44
- 210000001519 tissue Anatomy 0.000 description 21
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 18
- 241000699666 Mus <mouse, genus> Species 0.000 description 16
- 241000792859 Enema Species 0.000 description 15
- 239000007920 enema Substances 0.000 description 15
- 229940095399 enema Drugs 0.000 description 15
- 238000003304 gavage Methods 0.000 description 15
- 230000000968 intestinal effect Effects 0.000 description 15
- 239000000463 material Substances 0.000 description 15
- 230000037396 body weight Effects 0.000 description 10
- 230000008859 change Effects 0.000 description 10
- 238000007912 intraperitoneal administration Methods 0.000 description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- 230000001575 pathological effect Effects 0.000 description 9
- 238000011740 C57BL/6 mouse Methods 0.000 description 8
- 230000003110 anti-inflammatory effect Effects 0.000 description 8
- 206010009887 colitis Diseases 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- 210000002540 macrophage Anatomy 0.000 description 8
- 208000031648 Body Weight Changes Diseases 0.000 description 7
- 230000004579 body weight change Effects 0.000 description 7
- 208000002551 irritable bowel syndrome Diseases 0.000 description 7
- 238000011084 recovery Methods 0.000 description 7
- 208000037816 tissue injury Diseases 0.000 description 7
- 206010061218 Inflammation Diseases 0.000 description 6
- 239000003651 drinking water Substances 0.000 description 6
- 235000020188 drinking water Nutrition 0.000 description 6
- 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 5
- 108020004414 DNA Proteins 0.000 description 5
- 230000006378 damage Effects 0.000 description 5
- 235000019441 ethanol Nutrition 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- 230000006872 improvement Effects 0.000 description 5
- 230000004054 inflammatory process Effects 0.000 description 5
- 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 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 4
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 230000002757 inflammatory effect Effects 0.000 description 4
- 238000000465 moulding Methods 0.000 description 4
- 239000012890 simulated body fluid Substances 0.000 description 4
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 4
- 206010052360 Colorectal adenocarcinoma Diseases 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 125000002091 cationic group Chemical group 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007928 intraperitoneal injection Substances 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- 235000012239 silicon dioxide Nutrition 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 208000018522 Gastrointestinal disease Diseases 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 102000002689 Toll-like receptor Human genes 0.000 description 2
- 108020000411 Toll-like receptor Proteins 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 230000003064 anti-oxidating effect Effects 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- WOWHHFRSBJGXCM-UHFFFAOYSA-M cetyltrimethylammonium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+](C)(C)C WOWHHFRSBJGXCM-UHFFFAOYSA-M 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 239000003018 immunosuppressive agent Substances 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 210000004347 intestinal mucosa Anatomy 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000012044 organic layer Substances 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 229940054269 sodium pyruvate Drugs 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 210000001541 thymus gland Anatomy 0.000 description 2
- 230000036962 time dependent Effects 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- VQVUBYASAICPFU-UHFFFAOYSA-N (6'-acetyloxy-2',7'-dichloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) acetate Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(OC(C)=O)C=C1OC1=C2C=C(Cl)C(OC(=O)C)=C1 VQVUBYASAICPFU-UHFFFAOYSA-N 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- PQUCIEFHOVEZAU-UHFFFAOYSA-N Diammonium sulfite Chemical compound [NH4+].[NH4+].[O-]S([O-])=O PQUCIEFHOVEZAU-UHFFFAOYSA-N 0.000 description 1
- 206010061819 Disease recurrence Diseases 0.000 description 1
- 208000015877 Duodenal disease Diseases 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 208000034826 Genetic Predisposition to Disease Diseases 0.000 description 1
- 108010034143 Inflammasomes Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000001388 Opportunistic Infections Diseases 0.000 description 1
- ZYFVNVRFVHJEIU-UHFFFAOYSA-N PicoGreen Chemical compound CN(C)CCCN(CCCN(C)C)C1=CC(=CC2=[N+](C3=CC=CC=C3S2)C)C2=CC=CC=C2N1C1=CC=CC=C1 ZYFVNVRFVHJEIU-UHFFFAOYSA-N 0.000 description 1
- 210000000662 T-lymphocyte subset Anatomy 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- GUJOJGAPFQRJSV-UHFFFAOYSA-N dialuminum;dioxosilane;oxygen(2-);hydrate Chemical compound O.[O-2].[O-2].[O-2].[Al+3].[Al+3].O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O GUJOJGAPFQRJSV-UHFFFAOYSA-N 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 208000010643 digestive system disease Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 208000018595 duodenum disease Diseases 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 208000028299 esophageal disease Diseases 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000002479 food anti-caking agent Nutrition 0.000 description 1
- 208000018685 gastrointestinal system disease Diseases 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 229940124589 immunosuppressive drug Drugs 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910052901 montmorillonite Inorganic materials 0.000 description 1
- 239000005543 nano-size silicon particle Substances 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000008816 organ damage Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000004647 pro-inflammatory pathway Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003870 salicylic acids Chemical class 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000002336 sorption--desorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 208000018556 stomach disease Diseases 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
- A61K31/52—Purines, e.g. adenine
-
- 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
- A61K31/60—Salicylic acid; Derivatives thereof
- A61K31/606—Salicylic acid; Derivatives thereof having amino groups
-
- 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
- A61K31/655—Azo (—N=N—), diazo (=N2), azoxy (>N—O—N< or N(=O)—N<), azido (—N3) or diazoamino (—N=N—N<) compounds
-
- 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
- A61K47/02—Inorganic compounds
-
- 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
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive 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]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B33/00—Silicon; Compounds thereof
- C01B33/113—Silicon oxides; Hydrates thereof
- C01B33/12—Silica; Hydrates thereof, e.g. lepidoic silicic acid
- C01B33/18—Preparation of finely divided silica neither in sol nor in gel form; After-treatment thereof
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
- C07F7/1804—Compounds having Si-O-C linkages
- C07F7/1872—Preparation; Treatments not provided for in C07F7/20
- C07F7/1892—Preparation; Treatments not provided for in C07F7/20 by reactions not provided for in C07F7/1876 - C07F7/1888
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/01—Particle morphology depicted by an image
- C01P2004/03—Particle morphology depicted by an image obtained by SEM
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/01—Particle morphology depicted by an image
- C01P2004/04—Particle morphology depicted by an image obtained by TEM, STEM, STM or AFM
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/60—Particles characterised by their size
- C01P2004/64—Nanometer sized, i.e. from 1-100 nanometer
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2006/00—Physical properties of inorganic compounds
- C01P2006/12—Surface area
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2006/00—Physical properties of inorganic compounds
- C01P2006/16—Pore diameter
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Nanotechnology (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Crystallography & Structural Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Nutrition Science (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Rheumatology (AREA)
- Pain & Pain Management (AREA)
- Physiology (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a nano material capable of efficiently removing cfDNA and ROS, belonging to the field of pharmaceutical preparations. The preparation method of the nano material capable of efficiently removing cfDNA and ROS comprises the following steps: (1) preparing bis [3- (triethoxysilyl) propyl ] diselenide; (2) the preparation of the ROS nano material can be efficiently eliminated; (3) can effectively eliminate the preparation of cfDNA and ROS nano materials. The nano material capable of efficiently removing the cfDNA and the ROS, which is prepared by the invention, is the first nano material capable of simultaneously removing the cfDNA and the ROS, and the anti-inflammatory drug is loaded on the nano material capable of efficiently removing the cfDNA and the ROS, so that the effectiveness and the safety of the treatment of the inflammatory bowel disease can be effectively improved.
Description
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a nano material capable of efficiently removing cfDNA and ROS, and a preparation method and application thereof.
Background
Inflammatory bowel disease is a chronic, recurrent gastrointestinal disease caused by intestinal inflammation and injury, and mainly includes crohn's disease and ulcerative colitis. At present, the incidence rate of inflammatory bowel disease in developed countries is in a plateau stage, and no obvious improvement trend exists. With the obvious changes of the dietary structure, the life style and the living environment of the national people in China, the incidence rate of IBD (1.8/10 ten thousand people) is on a continuous and rapid increasing trend. At present, no specific medicine exists in clinical treatment of inflammatory bowel diseases, medicines such as salicylic acids, hormones, immunosuppressants, biological agents and the like are mainly used for treatment, but the traditional anti-inflammatory treatment medicines are single in effect, poor in curative effect and prone to disease recurrence. In addition, the frequent and long-term use of currently applied small molecule drugs or biologic-based immunosuppressive drugs may lead to systemic side effects and serious complications, including autoimmune diseases, opportunistic infections and organ damage. Therefore, on the basis of deeply clarifying the pathogenesis of inflammatory bowel disease, exploring new treatment strategies and developing new drugs are of great significance to the efficient and safe treatment of inflammatory bowel disease.
Immunotherapy is currently becoming an important strategy for the treatment of inflammatory bowel disease. A large number of researches show that genetic susceptibility and external environment stimulation cause the barrier function of normal intestinal mucosa to be disordered, the inherent immune cells enriched in the intestinal inherent layer are activated, excessive inflammatory reaction can cause the unbalance of immune cells such as macrophages, neutrophils, T lymphocyte subsets and the like, and further the inflammatory signal pathway is activated. At the same time, reactive oxygen species ROS and inflammasome generated during the inflammatory process accumulate in the intestinal epithelium causing oxidative tissue damage. In this process, the abnormal activation of Toll-like receptors on immune cells leads to the massive production of inflammatory mediators, which accelerates the occurrence and development of inflammatory diseases. Among them, free DNA (cell free DNA, cfDNA) has been shown not only to be a biomarker for the prognosis of IBD, but also to exacerbate the severity of inflammation and prolong the duration of inflammation by activating the pro-inflammatory pathway of immune cells, especially macrophage TLR 9. Based on the important role of dangerous molecules such as cfDNA and ROS in inflammatory bowel disease, the development of a material capable of rapidly removing cfDNA and ROS is expected to realize efficient treatment of inflammatory bowel disease.
Nanomedicine has very broad prospects in the diagnosis, prevention and treatment of various diseases. At present, dozens of nano-drugs are approved for clinical application, and hundreds of nano-drugs are in clinical trials. In the prevention and treatment of gastrointestinal diseases, the nano material also has unique advantages, such as the improvement of the stability of the medicine, the prevention of the degradation of the medicine, the improvement of the bioavailability, the improvement of the enrichment, the release and the retention of the medicine at the disease part, and the like. Among the nano materials, the nano silicon dioxide has high specific surface area and better biological safety, and is often used as an adsorbent for pollutant removal or used as a drug carrier for diagnosis and treatment of diseases. The silicon dioxide material is approved by the national food and drug administration as a medicinal auxiliary material or a food anti-caking agent and is widely used in the fields of food and drugs. In addition, oral administration of the traditional Chinese medicine montmorillonite powder is commonly used for adjuvant therapy of related pain symptoms caused by esophagus, stomach and duodenum diseases, and the main component of the oral administration also comprises silicon dioxide. Therefore, development of oral nanomaterials capable of removing cfDNA and ROS based on silica is expected to treat inflammatory bowel disease by removing dangerous molecules, and on this basis, improvement of therapeutic effect by carrying anti-inflammatory drugs is expected to be significant. To date, there has been no report on the treatment of inflammatory bowel disease by directly eliminating various dangerous molecules such as cfDNA and ROS through nanomaterials.
Disclosure of Invention
In order to overcome the defects and shortcomings of the prior art, the invention aims to provide a preparation method of a nano material capable of efficiently removing cfDNA and ROS.
The invention also aims to provide the nano-material which can effectively remove cfDNA and ROS and is prepared by the preparation method.
Still another object of the present invention is to provide the above application that can efficiently remove cfDNA and ROS nanomaterial.
The purpose of the invention is realized by the following technical scheme:
a preparation method of a nano material capable of efficiently removing cfDNA and ROS comprises the following steps:
(1) preparation of bis [3- (triethoxysilyl) propyl ] diselenide
Dripping gamma-chloropropyltrimethoxysilane into the sodium diselenide solution, stirring overnight, stopping reaction, extracting, drying, and purifying the crude product by chromatography to obtain a dark yellow liquid, namely bis [3- (triethoxysilyl) propyl ] diselenide;
(2) preparation of nano material capable of efficiently removing ROS
Dissolving a cation template agent and triethanolamine, heating and stirring; then adding a mixed solution of tetraethoxysilane, bis [3- (triethoxysilyl) propyl ] diselenide and ethanol, continuously stirring, centrifugally collecting nano particles, washing a solid crude product, refluxing an ammonium nitrate ethanol solution, centrifugally collecting, and drying to obtain a nano Material (MSN) capable of efficiently removing ROS;
(3) preparation of nano material capable of efficiently removing cfDNA and ROS
And (3) dispersing the nano Material (MSN) capable of efficiently removing the ROS in the step (2) into toluene, adding epoxypropyl trimethoxy silane to obtain a mixture, refluxing the mixture to obtain an epoxy modified nano material 1, centrifugally collecting, washing to obtain a solid crude product, adding the solid crude product into a polycation aqueous solution, reacting, centrifuging, and drying to obtain the nano material (MSN-PEI) capable of efficiently removing cfDNA and ROS.
The gamma-chloropropyltrimethoxysilane and the sodium diselenide in the step (1) are preferably calculated according to the molar ratio of 2: 1.
The time of the overnight stay in the step (1) is preferably 8-12 hours; more preferably 10 hours.
The reaction stop described in step (1) is preferably stopped by adding ice water.
The organic solvent used in the extraction described in step (1) is preferably dichloromethane.
The drying described in the step (1) is preferably carried out by drying the organic layer with anhydrous sodium sulfate.
The chromatography in step (1) is preferably performed by silica gel column chromatography; the solvent for silica gel column chromatography is preferably Petroleum Ether (PE) and Dichloromethane (DCM) of 10-1: 1.
The cationic template described in step (2) preferably includes at least one of cetyltrimethyl-p-toluenesulfonyl ammonium (CTAT), cetyltrimethyl ammonium chloride (CTAC), and cetyltrimethyl ammonium bromide (CTAB).
The cation template and triethanolamine in the step (2) are preferably calculated according to the mass ratio of 3-5: 1; more preferably in a mass ratio of 4: 1.
The dissolved solvent in step (2) is preferably water; more preferably ultrapure water.
The mass-volume ratio (g: mL) of the cationic template agent and the dissolved solvent in the step (2) is preferably 0.6: 35-45; more preferably in a mass to volume ratio (g: mL) of 0.6: 40.
In the heating and stirring in the step (2), the heating temperature is preferably 40-90 ℃; more preferably 80 deg.c.
In the heating and stirring in the step (2), the stirring time is preferably 0.5-2 hours; more preferably 1 hour.
In the heating and stirring in the step (2), the stirring speed is preferably 400-1200 rpm; more preferably 900 rpm.
In the mixed liquid of the ethyl orthosilicate, the bis [3- (triethoxysilyl) propyl ] diselenide and the ethanol in the step (2), the mass ratio of the ethyl orthosilicate to the bis [3- (triethoxysilyl) propyl ] diselenide is preferably 3-5: 1; more preferably in a mass ratio of 4: 1.
In the mixed solution of ethyl orthosilicate, bis [3- (triethoxysilyl) propyl ] diselenide and ethanol in the step (2), preferably calculating the bis [3- (triethoxysilyl) propyl ] diselenide and the ethanol according to the mass-to-volume ratio (g: mL) of 1: 1-3; more preferably 1:2 by mass to volume (g: mL).
The continuous stirring time in the step (2) is preferably 1-10 hours; more preferably 4 hours.
The washing in the step (2) is preferably washing with absolute ethyl alcohol; the number of washing is preferably at least three; more preferably three times.
The content of ammonium nitrate in the ammonium nitrate ethanol solution in the step (2) is 0-1 wt%.
The number of times of refluxing in the step (2) is preferably at least 3 times; more preferably three times.
The refluxing time in the step (2) is preferably 6-48 hours per time; more preferably 12 hours.
The drying in step (2) is preferably vacuum drying.
After the ROS nanometer material capable of efficiently removing the ROS is dispersed in the toluene in the step (3), the final concentration of the ROS nanometer material capable of efficiently removing the ROS is preferably 0.1-10 mg/mL; more preferably 2 mg/mL.
In the mixture in the step (3), the final concentration of the epoxypropyltrimethoxysilane is preferably 0.05-10 mg/mL; more preferably 1.5 mg/mL.
The refluxing condition in the step (3) is preferably 70-90 ℃ refluxing for 20-26 h; more preferably at 80 ℃ for 24 h.
The polycation in step (3) preferably comprises at least one of branched Polyethyleneimine (PEI) and hyperbranched Polyamidoamine (PAMAM).
In the polycation aqueous solution in the step (3), the concentration of the polycation is preferably 0.255-10 mg/mL; preferably 0.5 mg/mL.
The mass ratio of the solid crude product to the polycation in the step (3) is preferably 0.2: 0.5-1.5, and more preferably 0.2: 1.
The reaction condition in the step (3) is preferably room temperature reaction for 22-26 h; more preferably at 25 ℃ for 24 h.
A nano material capable of efficiently removing cfDNA and ROS is prepared by the preparation method.
The application of the nano material capable of efficiently removing cfDNA and ROS in preparing a medicament for treating and/or preventing inflammatory bowel disease is provided.
The inflammatory bowel disease includes but is not limited to at least one of mild acute inflammatory bowel disease, acute enteritis, severe acute inflammatory bowel disease, chronic inflammatory bowel disease and severe acute enteritis.
A medicine for treating and/or preventing inflammatory bowel disease comprises the nano-material capable of efficiently removing cfDNA and ROS.
The effective dose of the nano material capable of efficiently removing cfDNA and ROS is preferably 10-40 mg/kg.
The medicament for treating and/or preventing inflammatory bowel disease also comprises small molecule anti-inflammatory medicaments and/or derivatives of the small molecule anti-inflammatory medicaments.
The small-molecule anti-inflammatory drug preferably comprises at least one of mesalamine, sulfasalazine, azathioprine and methotrexate.
The administration form of the medicament for treating and/or preventing the inflammatory bowel disease comprises at least one of oral administration, gastric lavage, intraperitoneal injection and rectal enema.
The preparation method of the medicine for treating and/or preventing inflammatory bowel disease comprises the following steps:
respectively dissolving the micromolecular anti-inflammatory drug and/or the derivative of the micromolecular anti-inflammatory drug and the nano material capable of effectively removing cfDNA and ROS in an organic solvent, and mixing to obtain the compound.
The nano material capable of efficiently removing cfDNA and ROS and the micromolecular anti-inflammatory drug are preferably calculated according to the mass ratio of 1: 1-30; more preferably 1: 1.
The organic solvent preferably includes, but is not limited to, at least one of dimethyl sulfoxide (DMSO), water, glycerol, Dimethylformamide (DMF), and isopropanol; more preferably dimethyl sulfoxide (DMSO).
The mixing is preferably stirring mixing at room temperature; the rotating speed of the stirring is preferably 300-500 rpm; more preferably 400 rpm.
When the nano material is used for preparing a medicament for treating and/or preventing inflammatory bowel diseases, the cfDNA and ROS nano material can be efficiently removed to be used as a carrier for loading small-molecule anti-inflammatory medicaments and/or derivatives of the small-molecule anti-inflammatory medicaments.
Compared with the prior art, the invention has the following advantages and effects:
(1) the nano material capable of efficiently removing the cfDNA and the ROS, which is prepared by the invention, is the first nano material capable of simultaneously removing the cfDNA and the ROS, and the anti-inflammatory drug is loaded on the nano material capable of efficiently removing the cfDNA and the ROS, so that the effectiveness and the safety of the treatment of the inflammatory bowel disease can be effectively improved.
(2) The nano material capable of efficiently removing cfDNA and ROS can realize double functions of anti-inflammation and anti-oxidation through the function of efficiently removing cfDNA and ROS per se under the condition of not carrying any medicine, and realizes efficient and safe inflammatory bowel disease treatment.
(3) The nano material capable of efficiently removing cfDNA and ROS can accurately target inflammatory intestinal tissues under oral administration, and has retention for up to three days.
(4) The nano material capable of efficiently removing cfDNA and ROS prepared by the invention can be used as a long-acting preparation, has the same effect of treating inflammatory bowel diseases as daily administration under a high-dose low-frequency administration strategy, and has better biological safety.
(5) Compared with the clinical first-line use mesalazine, the nano material capable of efficiently removing cfDNA and ROS has better treatment effect on inflammatory bowel diseases and better biological safety.
(6) The nano material capable of efficiently removing cfDNA and ROS is used for loading small-molecule anti-inflammatory drugs and/or derivatives of the small-molecule anti-inflammatory drugs, so that degradation and drug release of ROS at a high level in a response inflammatory microenvironment are realized, and the treatment effect of inflammatory bowel diseases is further improved.
(7) The nano material capable of efficiently removing cfDNA and ROS prepared by the invention has the advantages of cheap and easily available raw materials, obvious effect of treating inflammatory bowel diseases and good clinical application prospect.
Drawings
FIG. 1 is a graph showing the results of the characterization of the ROS-scavenging nanomaterial (MSN) in example 1 of the present invention; wherein, the graph A is a scanning electron microscope imaging result graph capable of efficiently removing ROS nanometer Materials (MSN); FIG. B is a transmission electron microscope imaging result graph of ROS nanomaterial (MSN) can be efficiently eliminated; FIG. C is a graph of the pore size distribution results for ROS-scavenging nanomaterials (MSNs); and the graph D is a nitrogen isothermal adsorption and desorption curve chart of the ROS nanometer Material (MSN) capable of being efficiently removed.
FIG. 2 is a diagram of a situation in which the nanomaterial of the present invention in example 2 can efficiently scavenge cfDNA and ROS and scavenge cfDNA and ROS; wherein, the graph A is a transmission electron microscope imaging result graph of the ROS nanometer Material (MSN) capable of being efficiently cleared before incubation; FIG. B is a transmission electron microscope imaging result graph of cfDNA and ROS-scavenging nanomaterial (MSN-PEI) after incubation; and the graph C is a graph of the result of the binding rates of MSN-PEI, MSN and PEI and ct-DNA respectively with different mass ratios.
FIG. 3 is a graph of the anti-inflammatory and antioxidant damage at the cellular level that can efficiently scavenge cfDNA and ROS nanomaterials (MSN-PEI) in example 3 of the invention; wherein, the graph A is a TNF-alpha level result graph of co-incubation supernatant capable of efficiently removing cfDNA and ROS nano material (MSN-PEI); panel B is a graph of cell viability results for co-incubated cells that efficiently cleared cfDNA and ROS nanomaterial (MSN-PEI).
FIG. 4 is a graph of the prevention of DSS-induced mild acute inflammatory bowel disease by the nanomaterial capable of efficiently scavenging cfDNA and ROS in example 4 of the present invention; wherein, the graph A is a graph of the weight change result of the mice after 7 days of the gavage treatment; FIG. B is a graph showing the results of colon length changes in mice after 7 days of gavage treatment; FIG. C is a graph showing the results of pathological scoring of intestinal tissue sections of mice 7 days after the gavage treatment; FIG. D is a graph showing the results of body weight changes in mice after 7 days of intraperitoneal administration; FIG. E is a graph showing the results of colon length changes in mice after 7 days of intraperitoneal administration; FIG. F is a graph showing the results of pathological scoring of intestinal tissue sections of mice 7 days after intraperitoneal administration; FIG. G is a graph showing the results of body weight changes in mice after 7 days of rectal enema administration; FIG. H is a graph showing the results of colon length changes in mice after 7 days of rectal enema administration treatment; FIG. I is a graph showing the results of pathological scoring of intestinal tissue sections of mice 7 days after rectal enema administration.
FIG. 5 is a graph of the treatment of DSS-induced severe acute inflammatory bowel disease with nanomaterials that can efficiently scavenge cfDNA and ROS in example 5 of the present invention; wherein, the graph A is a graph of the weight change result of the mice within 14 days of the gavage treatment; panel B is a graph showing the results of colon length changes in mice after 14 days of gavage treatment; FIG. C is a graph showing the pathological scoring results of intestinal tissue sections of mice 14 days after the gavage treatment; FIG. D is a graph showing the results of the body weight change of the mice within 14 days of the intraperitoneal administration; FIG. E is a graph showing the results of colon length changes in mice after 14 days of intraperitoneal administration; FIG. F is a graph showing the pathological scoring results of intestinal tissue sections of mice 14 days after intraperitoneal administration; FIG. G is a graph showing the results of the body weight change of the mice within 14 days of the rectal enema administration treatment; FIG. H is a graph showing the results of colon length changes in mice 14 days after rectal enema administration treatment; FIG. I is a graph showing the results of pathological scoring of intestinal tissue sections of mice 14 days after rectal enema administration.
FIG. 6 is a graph of efficient elimination of cfDNA and ROS nanomaterials in example 6 of the invention for the treatment of TNBS-induced chronic inflammatory bowel disease; wherein, the graph A is a graph of the weight change result of the mice within 4 days of the gavage treatment; FIG. B is a graph showing the results of colon length changes in mice after 4 days of gavage treatment; FIG. C is a graph showing the results of pathological scoring of intestinal tissue sections of mice 4 days after the gavage treatment; FIG. D is a graph showing the results of body weight changes in mice within 4 days of intraperitoneal administration; FIG. E is a graph showing the results of colon length changes in mice after 4 days of intraperitoneal administration; FIG. F is a graph showing the pathological scoring results of intestinal tissue sections of mice after 4 days of intraperitoneal administration; FIG. G is a graph showing the results of body weight changes in mice within 4 days of rectal enema administration treatment; FIG. H is a graph showing the results of colon length changes in mice after 4 days of rectal enema administration treatment; FIG. I is a graph showing the results of pathological scoring of intestinal tissue sections of mice 4 days after rectal enema administration.
FIG. 7 is a graph of the biodistribution of the nanomaterials that can efficiently scavenge cfDNA and ROS in DSS-induced inflammatory bowel disease in example 7 of the present invention; wherein, the graph A is a result graph of the biodistribution of intestinal tissues after 2h, 6h, 12h, 24h, 48h and 72h of treatment by oral administration of the nano material capable of effectively removing cfDNA and ROS; and the graph B is a result graph of the fluorescence intensity quantification of intestinal tissues after 2h, 6h, 12h, 24h, 48h and 72h of treatment by oral administration of the nano material capable of effectively eliminating cfDNA and ROS.
FIG. 8 is a graph of the dose-dependent oral treatment of DSS-induced severe acute enteritis with nanomaterials that can efficiently scavenge cfDNA and ROS in example 8 of the present invention; wherein, the graph A is a graph of the weight change result of mice dosed with different doses of MSN-PEI through intragastric administration; FIG. B is a graph showing the effect of intragastric administration of different doses of MSN-PEI on colon length in mice; panel C is a graph of the effect of intragastric administration of different doses of MSN-PEI on the change in histological score in mice.
FIG. 9 is a graph of the time-dependent oral treatment of DSS-induced severe acute enteritis with efficient clearance of cfDNA and ROS nanomaterials in example 9 of the present invention; wherein, the graph A is a graph of the effect of the intragastric administration of 10mg/kg MSN-PEI on the body weight change of the mice at different administration intervals; FIG. B is a graph showing the effect of intragastric administration of 10mg/kg MSN-PEI on the colon length of mice at different administration intervals; FIG. C is a graph showing the effect of intragastric administration of 10mg/kg MSN-PEI on the change in histological score of mice at different administration intervals; FIG. D is a graph showing the effect of intragastric administration of 20mg/kg MSN-PEI on the change in body weight of mice at different dosing intervals; FIG. E is a graph showing the effect of intragastric administration of 20mg/kg MSN-PEI on the colon length of mice at different administration intervals; FIG. F is a graph showing the effect of intragastric administration of 20mg/kg MSN-PEI on the change in histological score of mice at different administration intervals; FIG. G is a graph showing the effect of intragastric administration of 40mg/kg MSN-PEI on the change in body weight of mice at different administration intervals; FIG. F is a graph showing the effect of intragastric administration of 40mg/kg MSN-PEI on the colon length of mice at different administration intervals; FIG. I is a graph showing the effect of intragastric administration of 40mg/kg MSN-PEI on the change in histological score of mice at different dosing intervals.
FIG. 10 is a graph showing the results of oral administration of the nanomaterial capable of efficiently eliminating cfDNA and ROS in treating DSS-induced severe acute enteritis in example 10 of the present invention; wherein, the graph A is a graph of the effect of different drugs and administration intervals on the body weight of the mice; panel B is a graph of the effect of different drugs and dosing intervals on colon length in mice; panel C is a graph of the effect of different drugs and dosing intervals on mouse histological scores.
FIG. 11 is a MSN-PEI supported mesalazine nano-drug containing 100 μ Μ H in example 11 of the present invention2O2Results of the release profile in simulated body fluid of (1).
FIG. 12 is a graph showing the results of oral administration of a MSN-PEI supported mesalazine nano-drug for treating DSS-induced inflammatory bowel disease according to example 12 of the present invention; wherein, the graph A is a result graph of the influence of the intragastric administration treatment of the MSN-PEI, mesalazine and MSN-PEI-loaded mesalazine nano-drug on the body weight of the mouse; FIG. B is a graph showing the effect of intragastric administration treatment of MSN-PEI, mesalazine and MSN-PEI-loaded mesalazine nano-drug on the length of the colon of a mouse; and the graph C is a result graph of the influence of the intragastric administration treatment of the MSN-PEI, mesalazine and MSN-PEI-loaded mesalazine nano-drug on the histological score of the mouse.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
In the present example, each reagent and raw material were commercially available unless otherwise specified.
Example 1: preparation of nano material capable of efficiently removing cfDNA and ROS
A preparation method of a nano material capable of efficiently removing cfDNA and ROS comprises the following steps:
(1) preparation of bis [3- (triethoxysilyl) propyl ] diselenide
Dropwise adding 10g of gamma-chloropropyltrimethoxysilane into 30mL of sodium diselenide solution (wherein the sodium diselenide solution contains 0.0255mol of sodium diselenide), stirring at room temperature (25 ℃) overnight (10 hours), adding ice water to stop reaction, extracting dichloromethane, drying an organic layer by using anhydrous sodium sulfate, and purifying a crude product by silica gel column (300-400 meshes) chromatography (petroleum ether (PE) and Dichloromethane (DCM) are 10-1: 1) to obtain a dark yellow liquid, namely bis [3- (triethoxysilyl) propyl ] diselenide;
(2) preparation of nano material capable of efficiently removing ROS
Taking 0.6g of cationic template agent (hexadecyl trimethyl p-toluene ammonium sulfonate) and 0.15g of triethanolamine, adding 40mL of ultrapure water into a round-bottom flask, fully dissolving, heating to 80 ℃, and stirring at 900rpm for 1 hour; then adding a mixed solution of tetraethoxysilane, bis [3- (triethoxysilyl) propyl ] diselenide and absolute ethyl alcohol, wherein: the mass of the tetraethoxysilane in the mixed solution is 4g, the mass of the bis [3- (triethoxysilyl) propyl ] diselenide is 1g, and the volume of the absolute ethyl alcohol is 2 mL; continuously stirring for 4 hours, collecting nano particles through centrifugation, and fully washing a solid crude product with absolute ethyl alcohol for three times; refluxing 1 wt% ammonium nitrate ethanol solution for 12 hours, centrifugally collecting, washing with ethanol for several times, repeatedly refluxing twice, and finally drying in vacuum to obtain the ROS (reactive oxygen species) removing nano Material (MSN) with high efficiency;
(3) preparation of nano material capable of efficiently removing cfDNA and ROS
Dispersing the ROS-efficiently-scavenging nano Material (MSN) obtained in the step (2) into toluene, wherein the final concentration of the ROS-efficiently-scavenging nano material is 2mg/mL, then adding epoxypropyltrimethoxysilane to obtain a mixture, wherein the final concentration of the epoxypropyltrimethoxysilane in the mixture is 1.5mg/mL, refluxing the mixture at 80 ℃ for 24 hours to obtain an epoxy modified nano material 1, centrifugally collecting nanoparticles, washing to obtain a solid crude product, ultrasonically dividing 0.2g of the solid crude product into 50mL of water, adding 2mL of a branched Polyethyleneimine (PEI) aqueous solution (the concentration of the branched polyethyleneimine in the branched polyethyleneimine aqueous solution is 0.5mg/mL), reacting at room temperature for 24 hours, centrifugally collecting, and drying to obtain the MSN-PEI capable of efficiently-scavenging cfDNA and ROS nano material.
The results are shown in FIG. 1, and the nitrogen adsorption-desorption curve represents that the diameter of the MSN nano-particles (capable of efficiently removing ROS nano-materials) is 60nm, the pore diameter is 4.2nm, and the specific surface area is 432.7m2·g-1。
Example 2: can effectively remove cfDNA and ROS (reactive oxygen species) nano material (MSN-PEI)
The nano material (MSN-PEI) capable of effectively eliminating cfDNA and ROS prepared in example 1 is added with 100 mu M H2O2The simulated body fluid (the simulated body fluid is purchased from Nanjing Yixun Biotechnology Ltd.) is incubated for 1 day, and the cfDNA and ROS nano-materials can be efficiently removed after the incubation is observed by a transmission electron microscope.
The results of transmission electron microscopy show that the MSN-PEI after incubation has ROS scavenging capability, and the results are shown in FIGS. 2A and 2B.
The ability of the ROS-scavenging nanomaterial (MSN), branched Polyethyleneimine (PEI) and MSN-PEI prepared in example 1 to bind to calf thymus DNA (ct-DNA) was evaluated. The ct-DNA solution (25. mu.L, 10. mu.g/mL, CAS number: 91080-16-9), PicoGreen (25. mu.L) and MilliQ water (50. mu.L) were mixed in the wells of a 96-well plate and incubated for 30min in the absence of light to form complexes. Then, MSN-PEI solution (100. mu.L) was added to the complex in a mass ratio (mg: mg) of MSN-PEI to calf thymus DNA (ct-DNA) of 0.1:1, 1:1, 10:1, 100:1, respectively. After incubation for 1h at 37 ℃, the fluorescence intensity of the complex after incubation at a wavelength of 520nm was measured with a multimode microplate reader (Molecular Devices, SpectraMax iD3) at a wavelength of 490 nm. At the same time, the MSN solution and PEI solution are used to replace the MSN-PEI solution to carry out the above test.
The nano-material (MSN-PEI) capable of efficiently eliminating cfDNA and ROS has high binding affinity with ct-DNA, and the result is shown in FIG. 2C.
Example 3: can effectively eliminate the conditions of anti-inflammation and anti-oxidation damage of cfDNA and ROS nano material at the cellular level
Evaluation of cfDNA and ROS scavenging Nanometric Material (MSN-PEI) prepared in example 1 with high efficiency in anti-inflammatory and antioxidant at cellular levelTo correct the condition of the injury. RAW 264.7 macrophages were purchased from American type culture Collection (ATCC, Manassas, Va.) and were maintained in DMEM containing 10% FBS, 1mM sodium pyruvate and 1% penicillin-streptomycin in a humidified environment at 37 deg.C (5% CO)2) And (5) culturing.
RAW 264.7 macrophages were administered at 2 × 104The density of individual cells/well was seeded in 96-well plates and LPS was added to a final concentration of 1. mu.g/mL for incubation for 30min to give activated RAW 264.7 mouse macrophages. Then co-incubating the activated RAW 264.7 mouse macrophage for 24h at 37 ℃ with an ROS nano material (MSN, final concentration of 10 mug/mL), branched polyethyleneimine (PEI, final concentration of 10 mug/mL) and a nano material (MSN-PEI, final concentration of 10 mug/mL) capable of efficiently removing cfDNA and ROS respectively, and collecting supernatant after completion to obtain an LPS + MSN group, an LPS + PEI group and an LPS + MSN-PEI group. Meanwhile, the Control group (LPS group) was prepared without any material, and the blank Control group (Control group) was prepared without any material and with an equal amount of physiological saline instead of LPS. And TNF-alpha levels of each group (LPS + MSN group, LPS + PEI group, and LPS + MSN-PEI group, Control group, and LPS group) were measured by ELISA kit (purchased from Shanghai Bohu Biotechnology Co., Ltd.) according to the instructions.
The test result shows that: the LPS + MSN group and the LPS + MSN-PEI group can effectively inhibit TNF-alpha secretion of macrophages of RAW 264.7 mice activated by LPS, the LPS + PEI group cannot inhibit TNF-alpha secretion of the macrophages, and the LPS + MSN group and the LPS + MSN-PEI group have good anti-inflammatory effects on cell water level, as shown in FIG. 3A.
Caco-2 human colorectal adenocarcinoma cells (purchased from ATCC) in DMEM containing 10% FBS, 1mM sodium pyruvate and 1% penicillin-streptomycin in a humidified environment at 37 ℃ (5% CO)2) And (5) culturing.
Caco-2 human colorectal adenocarcinoma cells at 2X 104Individual cells/well density were plated in 96-well plates (200. mu.L system) and H was added2O2(final concentration is 100 mu M), then adding a nano material (MSN, final concentration is 10 mu g/mL) capable of efficiently removing ROS and a nano material (MSN-PEI, final concentration is 10 mu g/mL) capable of efficiently removing cfDNA and ROS respectively, co-incubating at 37 ℃, discarding supernatant after 12H, and obtaining H2O2+MSGroup N and H2O2+ MSN-PEI group. While the control group (H) was prepared without adding any material2O2Group) to replace H with an equal amount of physiological saline without any material addition2O2Is a blank Control group (Control group). Each group (H)2O2+ MSN group, H2O2+ MSN-PEI group, Control group and H2O2Group) cells were treated with 2',7' -dichlorofluorescein diacetate (DCFH-DA, CAS No.: 4091-99-0) for 15min and washed 3 times with PBS buffer. Intracellular ROS production was analyzed by a Microplate Reader at 490nm excitation and 520nm emission wavelength.
The results show that H2O2+ MSN groups and H2O2The + MSN-PEI group can protect Caco-2 human colorectal adenocarcinoma cells from H2O2Mediated cytotoxic effects, H2O2+ MSN groups and H2O2The + MSN-PEI group all had antioxidant damage effect as shown in FIG. 3B.
Example 4: can effectively remove cfDNA and ROS nano-materials to prevent the condition of DSS-induced mild acute inflammatory bowel disease
Mice mild acute inflammatory bowel disease induced by dextran sulfate sodium salt (DSS) was treated prophylactically with the nano-material (MSN-PEI) that efficiently scavenges cfDNA and ROS of example 1. C57BL/6 mice (purchased from Beijing Wintolite laboratory animals technologies, Inc.) were classified into Control group, DSS model group, branched polyethyleneimine group (DSS + PEI, 10mg/kg), group capable of efficiently scavenging ROS nanomaterial (DSS + MSN, 10mg/kg), group capable of efficiently scavenging cfDNA and ROS nanomaterial (DSS + MSN-PEI, 10mg/kg), and 3% (w/v) DSS was continuously drunk for 7 days to establish colitis model in mice, and the Control group was given equivalent drinking water. The injection is administered by gavage, intraperitoneal injection or rectal enema for 7 days after the model building. Daily observations were recorded for mouse body weight and colon tissue was collected on day 7 to measure colon length, histopathological sections were taken, H & E stained and clinically scored.
The results are shown in FIGS. 4A-4I, indicating that: among three different administration modes, the nano material capable of efficiently removing cfDNA and ROS shows good IBD treatment effects in the aspects of weight, colon length recovery, colon tissue injury and the like, and is obviously superior to branched polyethyleneimine and the nano material capable of efficiently removing ROS. The nano-material can effectively eliminate cfDNA and ROS, and the prevention effect of the gavage delivery mode is the best among three administration modes.
Example 5: can effectively eliminate the condition that cfDNA and ROS nano-materials treat DSS-induced severe acute inflammatory bowel disease
Dextran sulfate sodium salt (DSS) -induced severe acute inflammatory bowel disease in mice was treated with the efficient removal of cfDNA and ROS nanomaterial (MSN-PEI) of example 1. The C57BL/6 mice are divided into a Control group, a DSS model group, a branched polyethyleneimine group (DSS + PEI, 10mg/kg), a ROS nanometer material group (DSS + MSN, 10mg/kg) capable of efficiently removing, cfDNA and ROS nanometer material group (DSS + MSN-PEI, 10mg/kg), 3% (w/v) DSS is continuously drunk for 7 days to establish a mouse colitis model, and the Control group gives equal amount of drinking water. The injection is administered by gavage or intraperitoneal injection or rectal enema for 11 consecutive days three days after the model building. Daily observations were recorded for mouse body weight and colon tissue was collected on day 14 to measure colon length, histopathological sections were taken, H & E stained and clinically scored.
The results are shown in FIGS. 5A-5I, indicating that: among three different administration modes, the nano material capable of efficiently removing cfDNA and ROS shows good IBD treatment effects in the aspects of weight, colon length recovery, colon tissue injury and the like, and is obviously superior to branched polyethyleneimine and the nano material capable of efficiently removing ROS. The nano material can effectively eliminate cfDNA and ROS and has the best treatment effect on the gastric lavage delivery nano material in three administration modes for treating the severe acute inflammatory bowel disease.
Example 6: can effectively eliminate the condition that cfDNA and ROS nano-materials are used for treating TNBS-induced chronic inflammatory bowel disease
The nano-material (MSN-PEI) capable of efficiently removing cfDNA and ROS in example 2 is adopted to treat the mouse chronic inflammatory bowel disease induced by trinitrobenzenesulfonic acid (TNBS). The C57BL/6 mice are divided into a Control group, a dextran sulfate sodium salt (DSS, MW: 36000-50000) model group, a branched polyethyleneimine group (DSS + PEI, 10mg/kg), a group capable of efficiently removing ROS nano-materials (DSS + MSN, 10mg/kg), a group capable of efficiently removing cfDNA and ROS nano-materials (DSS + MSN-PEI, 10mg/kg), 2.5% (w/v) TNBS is given to rectal enema to establish a mouse chronic colitis model, and the Control group is given with the same amount of physiological saline. Mice were observed daily for 4 consecutive days post-molding for gavage or intraperitoneal or rectal enema administration, and colon tissue was collected on day 4 to measure colon length, histopathological sections were performed, H & E stained and scored clinically.
The results are shown in FIGS. 6A-6I, indicating that: among three different administration modes, the nano material capable of efficiently removing cfDNA and ROS shows good IBD treatment effects in the aspects of weight, colon length recovery, colon tissue injury and the like, and is obviously superior to branched polyethyleneimine and ROS removing nano material. The nano material for rectal enema delivery has the best treatment effect among three administration modes for effectively eliminating cfDNA and ROS nano materials to treat chronic inflammatory bowel diseases.
Example 7: can effectively eliminate the biodistribution of cfDNA and ROS nano materials in a DSS-induced acute enteritis model
The biodistribution of cfDNA and ROS nanomaterial (MSN-PEI) of example 1 in a DSS-induced acute enteritis model was used to efficiently clear. The C57BL/6 mice are divided into a Cy 7-labeled branched polyethyleneimine group (PEI, 10mg/kg), a Cy 7-labeled group capable of efficiently scavenging ROS (MSN, 10mg/kg), and a Cy 7-labeled group capable of efficiently scavenging cfDNA and ROS (MSN-PEI, 10 mg/kg). Cy7-NHS was used to prepare dye-labeled MSN, PEI and MSN-PEI. The specific synthetic process comprises the following steps: 1mg of Cy7-NHS was mixed with 10mg of MSN, PEI or MSN-PEI, shaken overnight at 4 ℃ and centrifuged to remove unreacted dye molecules to obtain Cy 7-labeled MSN, Cy 7-labeled PEI and Cy 7-labeled MSN-PEI. After 3% (w/v) DSS was continuously drunk for 3 days to establish a mouse colitis model and treated for 2h, 6h, 12h, 24h, 48h and 72h by single oral administration, intestinal tissues were photographed and fluorescence intensity quantified using an NIR imaging system (IVIS Spectrum, Caliper Life) Cy7 filter channel.
The results are shown in FIGS. 7A-7B, indicating that: the oral administration can effectively remove cfDNA and ROS nano material, obviously increases fluorescence intensity in inflamed colon tissues, shows obvious retention effect, and is superior to branched polyethyleneimine and ROS nano material.
Example 8: can effectively eliminate the condition of severe acute enteritis induced by DSS (digital signal processing) through nano-material dose-dependent oral administration of cfDNA (deoxyribonucleic acid) and ROS (reactive oxygen species)
The cfDNA and ROS scavenging nanomaterial (MSN-PEI) of example 1 with high efficiency was used to treat severe acute enteritis. The C57BL/6 mice were divided into a Control group, a DSS model group, and MSN-PEI groups (2.5mg/kg, 5mg/kg, 10mg/kg, 20mg/kg, 40mg/kg) at different doses, and 3% (w/v) DSS was continuously drunk for 7 days to establish a colitis model of the mice, and the Control group was given an equivalent amount of drinking water. Intragastric administration is carried out for 11 consecutive days three days after the molding. Daily observations were recorded for mouse body weight and colon tissue was collected on day 14 to measure colon length, histopathological sections were taken, H & E stained and clinically scored.
The results are shown in FIGS. 8A-8C, indicating that: the nano material capable of efficiently removing cfDNA and ROS shows dose dependence on the treatment effects of weight, colon length recovery, colon tissue injury and the like. When the MSN-PEI is administrated at the dose of 10mg/kg, 20mg/kg and 40mg/kg, the MSN-PEI has good treatment effect, wherein the treatment effect of 40mg/kg is the best.
Example 9: can effectively eliminate the condition of severe acute enteritis induced by DSS through the time-dependent oral treatment of cfDNA and ROS nano material
The cfDNA and ROS scavenging nanomaterial of example 1 (MSN-PEI) was used to treat severe acute enteritis at different dosing intervals. The C57BL/6 mice were divided into a Control group, a DSS model group, a 10mg/kg MSN-PEI group (E1, E2), a 20mg/kg MSN-PEI group (E1, E2, E3), and a 40mg/kg MSN-PEI group (E1, E2, E3), and 3% (w/v) DSS was continuously drunk for 7 days to establish a colitis model of the mice, and the Control group was given an equal amount of drinking water. E1, E2, E3 indicate daily administration, every other day administration and every third day administration, respectively. The injection is administered by intragastric administration at different intervals for 11 days continuously after three days of molding. Daily observations were recorded for mouse body weight and colon tissue was collected on day 14 to measure colon length, histopathological sections were taken, H & E stained and clinically scored.
The results are shown in FIGS. 9A-9I, indicating that: when the administration intervals are changed under different dosages, the cfDNA and ROS nano-materials can be effectively eliminated, the treatment effect is good in the aspects of the treatment effects of weight, colon length recovery, colon tissue injury and the like, and the more the administration times are under the same dosage, the better the treatment effect is. When the administration dose was increased to 40mg/kg, similar therapeutic effects were exhibited at different administration intervals.
Example 10: can effectively eliminate the condition that cfDNA and ROS nano-material are orally taken to treat severe acute enteritis induced by DSS
The cfDNA and ROS scavenging nanomaterial of example 1 (MSN-PEI) was used to treat severe acute enteritis at different dosing intervals. The C57BL/6 mice were divided into a Control group, a DSS model group, a mesalazine (5-ASA) group (E1) at 10mg/kg, a mesalazine group (E3) at 40mg/kg, an MSN-PEI group (E1) at 10mg/kg, and an MSN-PEI group (E3) at 40mg/kg, and 3% (w/v) DSS was continuously drunk for 7 days to establish a colitis model of the mice, and the Control group was given an equivalent amount of drinking water. E1, E3 indicate daily and every other day, respectively. Gavage administration is carried out for 5 days after molding and for 16 days continuously according to different intervals. Daily observations were recorded for mouse body weight and colon tissue was collected on day 14 to measure colon length, histopathological sections were taken, H & E stained and clinically scored.
The results are shown in FIGS. 10A-10C, indicating that: the MSN-PEI with the dosage of 10mg/kg per day has good treatment effects on the aspects of weight and colon length recovery, colon tissue injury and other treatment effects, and has the effect equivalent to that of the positive drug mesalazine; the MSN-PEI with the dosage of 40mg/kg is given every two days, the treatment effect is remarkably superior to that of mesalazine, and the nano material capable of efficiently removing cfDNA and ROS has a long-acting effect on severe acute enteritis.
Example 11: preparation of multifunctional nano-drug and ROS-responsive drug release profile
Based on the nano material (MSN-PEI) capable of efficiently removing cfDNA and ROS obtained in the example 1, the nano material is ultrasonically dispersed into dimethyl sulfoxide (DMSO), and the final concentration of the MSN-PEI is 2 mg/mL; and adding mesalamine dissolved in DMSO to obtain a final concentration of the mesalamine dissolved in the DMSO of 2mg/mL, and stirring at room temperature (400rpm) for 12 hours to prepare the MSN-PEI loaded mesalamine nano-drug. At 37 deg.C, containing 100 μm H2O2Is dialyzed for 1 day at predetermined time points (1h, 2h, 4h, 8h, 16h, 24 h)h) Adding 1mL of dialysis external solution into the same volume of solution containing 100 μm H2O2Measuring the ultraviolet absorption intensity at 298nm wavelength, and detecting the ROS response to the drug release. With H2O substitution of H in simulated body fluid2O2The above test was performed as a control group.
The results show that the MSN-PEI supported mesalazine nano-drug has ROS-responsive drug release, and the results are shown in FIG. 11.
Example 12: situation of oral treatment of DSS-induced acute enteritis by MSN-PEI (Mesalazine-loaded) nano-drug
The MSN-PEI supported mesalazine nano-drug of example 11 was used for the prophylactic treatment of DSS-induced acute enteritis in mice. The C57BL/6 mice are divided into a Control group, a DSS model group, a mesalazine group of 10mg/kg, MSN-PEI of 10mg/kg and MSN-PEI-loaded mesalazine group of 10mg/kg, 3% (w/v) DSS is continuously drunk for 7 days to establish a colitis model of the mice, and the Control group is given equal amount of drinking water. The administration is carried out by gavage for 7 days. Daily observations were recorded for mouse body weight and colon tissue was collected on day 7 to measure colon length, histopathological sections were taken, H & E stained and clinically scored.
The results are shown in fig. 12A-12C, which indicate that: each group shows good IBD treatment effect in the aspects of weight, colon length recovery, colon tissue injury and the like, and the treatment effect of MSN-PEI (mesalamine) -loaded is obviously superior to that of the other two groups. The nano material can effectively remove cfDNA and ROS and can be used as a drug carrier to play a role in multifunctional IBD treatment.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of a nano material capable of efficiently removing cfDNA and ROS is characterized by comprising the following steps:
(1) preparation of bis [3- (triethoxysilyl) propyl ] diselenide
Dripping gamma-chloropropyltrimethoxysilane into the sodium diselenide solution, stirring at room temperature overnight, stopping reaction, extracting, drying, and purifying the crude product by chromatography to obtain a dark yellow liquid, namely bis [3- (triethoxysilyl) propyl ] diselenide;
(2) preparation of nano material capable of efficiently removing ROS
Dissolving a cation template agent and triethanolamine, heating and stirring; then adding a mixed solution of tetraethoxysilane, bis [3- (triethoxysilyl) propyl ] diselenide and ethanol, continuously stirring, centrifugally collecting nano particles, washing a solid crude product, refluxing an ammonium nitrate ethanol solution, centrifugally collecting, and drying to obtain the nano material capable of efficiently removing the ROS;
(3) preparation of nano material capable of efficiently removing cfDNA and ROS
Dispersing the nano material capable of efficiently removing the ROS in the step (2) in toluene, adding epoxypropyl trimethoxy silane to obtain a mixture, refluxing the mixture to obtain an epoxy modified nano material 1, centrifugally collecting, washing to obtain a solid crude product, adding the solid crude product into a polycation aqueous solution, reacting, centrifuging, and drying to obtain the nano material capable of efficiently removing cfDNA and ROS.
2. The method of claim 1, wherein the polycation in the step (3) comprises at least one of a branched polyethyleneimine and a hyperbranched polyamidoamine;
in the polycation aqueous solution in the step (3), the concentration of the polycation is 0.255-10 mg/mL.
3. A nanomaterial capable of efficiently removing cfDNA and ROS, characterized by being prepared by the preparation method of any one of claims 1-2.
4. The use of the nanomaterial capable of efficiently scavenging cfDNA and ROS of claim 3 in the preparation of a medicament for treating and/or preventing inflammatory bowel disease.
5. The use of claim 4, wherein the inflammatory bowel disease comprises, but is not limited to, at least one of mild acute inflammatory bowel disease, acute enteritis, severe acute inflammatory bowel disease, chronic inflammatory bowel disease, and severe acute enteritis.
6. A medicament for treating and/or preventing inflammatory bowel disease, comprising the efficient clearance cfDNA and ROS nanomaterial of claim 3.
7. The drug for treating and/or preventing inflammatory bowel disease according to claim 6, wherein the effective dose for efficiently eliminating cfDNA and ROS nano-materials is 10-40 mg/kg.
8. The agent for the treatment and/or prevention of inflammatory bowel disease according to claim 6, wherein said agent for the treatment and/or prevention of inflammatory bowel disease further comprises a small molecule anti-inflammatory agent, and/or a derivative of a small molecule anti-inflammatory agent.
9. The agent for the treatment and/or prevention of inflammatory bowel disease according to claim 8, wherein said small molecule anti-inflammatory agent includes but is not limited to at least one of mesalamine, sulfasalazine, azathioprine, and methotrexate.
10. The method for producing a medicament for treating and/or preventing inflammatory bowel disease according to any one of claims 6 to 9, characterized by comprising the steps of:
respectively dissolving the nano material capable of efficiently removing cfDNA and ROS and the micromolecule anti-inflammatory drug and/or the derivative of the micromolecule anti-inflammatory drug in an organic solvent, and mixing to obtain the compound preparation;
wherein the nano material capable of efficiently removing cfDNA and ROS and the micromolecular anti-inflammatory drug are calculated according to the mass ratio of 1: 1-30;
the organic solvent includes, but is not limited to, at least one of dimethyl sulfoxide, water, glycerol, dimethylformamide and isopropanol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111425453.5A CN114224911A (en) | 2021-11-26 | 2021-11-26 | Nano material capable of efficiently removing cfDNA and ROS, and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111425453.5A CN114224911A (en) | 2021-11-26 | 2021-11-26 | Nano material capable of efficiently removing cfDNA and ROS, and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114224911A true CN114224911A (en) | 2022-03-25 |
Family
ID=80751592
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111425453.5A Pending CN114224911A (en) | 2021-11-26 | 2021-11-26 | Nano material capable of efficiently removing cfDNA and ROS, and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114224911A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114712326A (en) * | 2022-04-15 | 2022-07-08 | 华南理工大学 | Antibiotic-loaded cationic mesoporous silica nanomaterial and preparation method and application thereof |
| CN117110596A (en) * | 2023-09-06 | 2023-11-24 | 方舟生物安全科技(广州)有限公司 | Modified glass slide for enriching cfDNA, method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112057434A (en) * | 2020-08-26 | 2020-12-11 | 华南理工大学 | Mesoporous silicon nano particle capable of responding X-ray medicine release and preparation method and application thereof |
| CN112236529A (en) * | 2018-06-01 | 2021-01-15 | 健诺心理股份有限公司 | Method for detecting unstable cell-free DNA and apparatus using the same |
| WO2021218763A1 (en) * | 2020-04-30 | 2021-11-04 | 中国科学院上海药物研究所 | Periploside composition, preparation method therefor and use thereof in preparation of drug for treatment of autoimmune diseases |
-
2021
- 2021-11-26 CN CN202111425453.5A patent/CN114224911A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112236529A (en) * | 2018-06-01 | 2021-01-15 | 健诺心理股份有限公司 | Method for detecting unstable cell-free DNA and apparatus using the same |
| WO2021218763A1 (en) * | 2020-04-30 | 2021-11-04 | 中国科学院上海药物研究所 | Periploside composition, preparation method therefor and use thereof in preparation of drug for treatment of autoimmune diseases |
| CN112057434A (en) * | 2020-08-26 | 2020-12-11 | 华南理工大学 | Mesoporous silicon nano particle capable of responding X-ray medicine release and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| JIANATI DAWULIETI等: "Treatment of severe sepsis with nanoparticulate cell-free DNA scavengers", 《SCIENCE ADVANCES》 * |
| 崔天国 等, 河南科学技术出版社 * |
| 张立德 等: "《超微粉体制备与应用技术》", 30 April 2001, 中国石化出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114712326A (en) * | 2022-04-15 | 2022-07-08 | 华南理工大学 | Antibiotic-loaded cationic mesoporous silica nanomaterial and preparation method and application thereof |
| CN114712326B (en) * | 2022-04-15 | 2023-09-22 | 华南理工大学 | Antibiotic-loaded cationic mesoporous silica nanomaterial and preparation method and application thereof |
| CN117110596A (en) * | 2023-09-06 | 2023-11-24 | 方舟生物安全科技(广州)有限公司 | Modified glass slide for enriching cfDNA, method and application thereof |
| CN117110596B (en) * | 2023-09-06 | 2024-01-26 | 方舟生物安全科技(广州)有限公司 | Modified glass slide for enriching cfDNA, method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114224911A (en) | Nano material capable of efficiently removing cfDNA and ROS, and preparation method and application thereof | |
| CA2476834C (en) | Natural cyclodextrin complexes of cannabinoids | |
| Xu et al. | pH/ROS dual-sensitive and chondroitin sulfate wrapped poly (β-amino ester)-SA-PAPE copolymer nanoparticles for macrophage-targeted oral therapy for ulcerative colitis | |
| CN102060991B (en) | Amphiphilic prodrug of 7- ethyl-10-hydroxycamptothecin and preparation method thereof | |
| WO2017162108A1 (en) | Pillararene complex, preparation method, pharmaceutical composition and use thereof | |
| Zhao et al. | Berberine-loaded carboxylmethyl chitosan nanoparticles ameliorate DSS-induced colitis and remodel gut microbiota in mice | |
| CN104491868B (en) | It is new to be based on antibody coupling chemotherapeutics nanometer ADC and preparation method and application | |
| Li et al. | Improvement of pneumonia by curcumin-loaded bionanosystems based on platycodon grandiflorum polysaccharides via calming cytokine storm | |
| CN104004085A (en) | Mycobacterium tuberculosis LAM oligosaccharide conjugate as well as preparation method and application thereof | |
| CN103705943A (en) | Preparation method and application of reduction-response-type pegylation (PEG) nanomedicine composition | |
| CN103705534A (en) | Preparation of natural active substance constructed polymer composite medicine and application thereof in inhibiting angiogenesis | |
| KR20170125994A (en) | Pharmaceutical composition containing silibinin and Puerariae Radix extract | |
| CN114917362B (en) | Lipid nanoparticle and preparation method and application thereof | |
| CN108148193B (en) | Cholic acid-containing high polymer material and liposome modified by same | |
| Huang et al. | Hydrogen peroxide responsive covalent cyclodextrin framework for targeted therapy of inflammatory bowel disease | |
| CN107049944B (en) | Polymer micelle capable of realizing simultaneous administration of sorafenib and curcumin and preparation method thereof | |
| CN104045823B (en) | A kind of Enoxolone derivative and its preparation method and application | |
| CN111793147B (en) | Modified chitosan, double-response nano-carrier drug, and preparation method and application thereof | |
| BRPI0716890A2 (en) | COMPOSITION, AND METHOD OF PRODUCTION OF A COMPOSITION, ADMINISTRATION OF A PHARMACEUTICALLY ACTIVE WATER INSOLUBLE AGENT TO A MAMMER, AND CANCER TREATMENT IN A MAMMER | |
| WO2008028336A1 (en) | A pharmaceutical composition of scutellarein and scutellarin or baicalin having synergistical antitumor effect | |
| CN105879051A (en) | Preparation and application of self-assembled nano-drug of core-shell structure | |
| Hu et al. | Salvianolic acid B-loaded polydopamine-modified hollow mesoporous organic silica nanoparticles for treatment of breast cancer metastasis via suppressing cancer-associated fibroblasts | |
| CN102652834A (en) | Preparation method and composite of low-molecular heparin nanopolymer | |
| CN112791189B (en) | High water-solubility 5-aminosalicylic acid pharmaceutical composition and preparation, preparation and application thereof | |
| WO2011044824A1 (en) | Inclusion complex of deoxypodophyllotoxin of cyclodextrin, preparation method, use for treament of cancer thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220325 |