CN116159039A - Near infrared light response conjugated polymer/phage composite nanoparticle for destroying biological film - Google Patents
Near infrared light response conjugated polymer/phage composite nanoparticle for destroying biological film Download PDFInfo
- Publication number
- CN116159039A CN116159039A CN202310142147.3A CN202310142147A CN116159039A CN 116159039 A CN116159039 A CN 116159039A CN 202310142147 A CN202310142147 A CN 202310142147A CN 116159039 A CN116159039 A CN 116159039A
- Authority
- CN
- China
- Prior art keywords
- nanoparticle
- bacterial
- phage
- temperature
- conjugated polymer
- 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
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 69
- 229920000547 conjugated polymer Polymers 0.000 title claims abstract description 31
- 239000002131 composite material Substances 0.000 title abstract description 8
- 230000004298 light response Effects 0.000 title abstract description 7
- 230000001580 bacterial effect Effects 0.000 claims abstract description 45
- 239000003814 drug Substances 0.000 claims abstract description 31
- 229940079593 drug Drugs 0.000 claims abstract description 16
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 11
- 241000894006 Bacteria Species 0.000 claims description 27
- 238000000034 method Methods 0.000 claims description 19
- 230000000694 effects Effects 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 11
- 239000000725 suspension Substances 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 8
- 208000015181 infectious disease Diseases 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 5
- 210000000170 cell membrane Anatomy 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 229920000642 polymer Polymers 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 4
- 230000003213 activating effect Effects 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000000969 carrier Substances 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 229940126586 small molecule drug Drugs 0.000 claims description 2
- 150000003384 small molecules Chemical class 0.000 claims description 2
- 241001515965 unidentified phage Species 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 230000008859 change Effects 0.000 abstract description 8
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 12
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 8
- 206010059866 Drug resistance Diseases 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 6
- TUJWIYZCAPMHSA-UHFFFAOYSA-N dipentylphosphoryloxybenzene Chemical compound CCCCCP(=O)(CCCCC)OC1=CC=CC=C1 TUJWIYZCAPMHSA-UHFFFAOYSA-N 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- KILNVBDSWZSGLL-KXQOOQHDSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCC KILNVBDSWZSGLL-KXQOOQHDSA-N 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- GSDSWSVVBLHKDQ-JTQLQIEISA-N Levofloxacin Chemical compound C([C@@H](N1C2=C(C(C(C(O)=O)=C1)=O)C=C1F)C)OC2=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-JTQLQIEISA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000002147 killing effect Effects 0.000 description 4
- 229960003376 levofloxacin Drugs 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 239000012221 photothermal agent Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- HXEACLLIILLPRG-YFKPBYRVSA-N L-pipecolic acid Chemical compound [O-]C(=O)[C@@H]1CCCC[NH2+]1 HXEACLLIILLPRG-YFKPBYRVSA-N 0.000 description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 244000172533 Viola sororia Species 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 239000004599 antimicrobial Substances 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000032770 biofilm formation Effects 0.000 description 2
- 229960003405 ciprofloxacin Drugs 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000000254 damaging effect Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000002296 dynamic light scattering Methods 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- HXEACLLIILLPRG-RXMQYKEDSA-N l-pipecolic acid Natural products OC(=O)[C@H]1CCCCN1 HXEACLLIILLPRG-RXMQYKEDSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 2
- 229960002930 sirolimus Drugs 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- -1 3, 6-bis (5-bromothiophen-2-yl) -2, 5-di (octyldodecyl) pyrrolo [3,4-c]Pyrrole-1, 4-dione Chemical compound 0.000 description 1
- ROFVEXUMMXZLPA-UHFFFAOYSA-N Bipyridyl Chemical compound N1=CC=CC=C1C1=CC=CC=N1 ROFVEXUMMXZLPA-UHFFFAOYSA-N 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 102000003844 DNA helicases Human genes 0.000 description 1
- 108090000133 DNA helicases Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 102000004310 Ion Channels Human genes 0.000 description 1
- 239000007987 MES buffer Substances 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 1
- 235000005811 Viola adunca Nutrition 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 235000002254 Viola papilionacea Nutrition 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 238000003570 cell viability assay Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000003235 crystal violet staining Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000004768 lowest unoccupied molecular orbital Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000007669 thermal treatment Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Images
Classifications
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5192—Processes
-
- 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
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/436—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having oxygen as a ring hetero atom, e.g. rapamycin
-
- 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
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
-
- 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
- A61K31/4985—Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
-
- 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
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5383—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with heterocyclic ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5123—Organic compounds, e.g. fats, sugars
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5176—Compounds of unknown constitution, e.g. material from plants or animals
- A61K9/5184—Virus capsids or envelopes enclosing drugs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a near infrared light response conjugated polymer/phage composite nanoparticle for destroying bacterial biofilms. The nanoparticle is formed by autonomous loading of amphiphilic molecules, conjugated polymers, small molecular drugs and phage. According to the invention, the outer layer of the nanoparticle is modified with phage, the bacterial biofilm is precisely positioned by the phage, the conjugated polymer converts light energy into heat energy, and the phase change of amphiphilic molecules is initiated to release internal hydrophobic drugs, so that the bacterial biofilm is precisely and controllably damaged in time and space.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a near infrared light response conjugated polymer/phage composite nanoparticle for destroying a bacterial biofilm, which comprises preparation of the nanoparticle and multifunctional research for destroying the biofilm.
Background
The biological membrane is formed by gathering bacteria and extracellular matrixes such as polysaccharide, protein, extracellular DNA and the like secreted by the bacteria. The biofilm matrix acts as a protective barrier so that the drug cannot penetrate the full depth of the biofilm, resulting in difficulty in achieving damage to the biofilm by the antimicrobial drug. Bacteria within the biofilm are in a slow growing and starving state with reduced sensitivity to antimicrobial agents. In addition, bacteria wrapped by the biological film carry out the horizontal transfer of drug resistance genes, so that the drug resistance of the bacteria is enhanced. The drug resistance of the biomembrane to the antibacterial drug is 10-1000 times that of planktonic bacteria, which not only increases the difficulty of treating bacterial infection, but also further increases the possibility of generating drug resistance of bacteria. Therefore, it is very important to develop new therapeutic means that can effectively destroy the biofilm and avoid developing drug resistance.
In recent years, light-operated treatment modes have attracted extensive interest in developing novel drug carriers and treatment modes due to their better spatial and temporal controllability. The near infrared light has strong tissue penetrating capacity and small damage to tissues, can accurately reach deep tissues of a target without causing side reactions, so that a light control treatment mode based on the near infrared light becomes a research hot point of the light control treatment mode.
Among the photo-thermal agents with near infrared response, the conjugated polymer has excellent photo-thermal conversion efficiency and optical stability, and overcomes the defects of poor stability and biocompatibility of the traditional inorganic photo-thermal agent. The plurality of light absorbing units of the conjugated polymer can achieve a stronger light absorbing capacity and signal amplifying effect than the organic small molecule photothermal agent. Therefore, constructing photo-thermal nanoparticles based on conjugated polymers of near infrared light response for photo-thermal treatment and drug delivery shows great application value.
At present, nanoparticles based on near infrared light response are mainly focused on utilizing the photothermal effect of the nanoparticles to thermally damage bacteria infection sites or tissues, so that the aim of killing bacteria is fulfilled. However, such nanoparticles have poor specific selective action on pathogenic bacteria and are limited to achieving space-time controllable irreversible damage to bacteria and limited destructive action on bacterial biofilms.
Disclosure of Invention
In view of the defects of the existing method for destroying bacteria and biological membranes thereof, the invention provides the near infrared light response conjugated polymer/phage composite nanoparticle which can rapidly heat up under the irradiation of near infrared light and directionally release drugs, thereby achieving the effect of destroying the biological membranes.
The nanoparticle provided by the invention is formed by autonomous assembly of amphiphilic molecules, conjugated polymers, small molecular drugs and bacteriophage;
the nano particles are in a spherical shape, the particle size is 5-500 nm, and the particle size can be 20-300nm;
the amphiphilic molecules comprise temperature-responsive amphiphilic molecules and non-temperature-sensitive amphiphilic molecules;
the temperature-responsive amphiphilic molecules are molecules with obvious state change at different temperatures, can generally undergo phase change at 30-55 ℃, are converted from a solid state to a molten state, and can be specifically dipalmitoyl phosphatidylcholine (DPPC), and the phase change temperature is about 38 ℃;
the non-temperature sensitive amphipathy is a molecule with insignificant change of hydrophilicity and hydrophobicity at different temperatures, and can be specifically distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG);
the conjugated polymer is provided with an aromatic conjugated main chain and an aliphatic side chain, wherein the conjugated main chain is formed by copolymerizing an acceptor conjugated unit (the electron withdrawing capability is strong, the lowest unoccupied molecular orbital energy level is low) and a donor conjugated unit (the electron donating capability is strong, and the highest occupied orbital energy level is high); the highest absorption peak of conjugated polymer molecule is 600nm-1200 nm; the polymer molecules have the capability of converting absorbed near infrared light into heat, and the energy conversion efficiency is not lower than 30 percent, namely, the polymer molecules have stronger photo-thermal effect and photo-thermal conversion efficiency;
the polymer conjugated molecules include, but are not limited to, the following structures:
the small molecular medicine is a hydrophobic medicine, and the target point of the small molecular medicine is bacterial cell membrane or biological molecules outside the cell membrane, including but not limited to cell membrane, ion channel, intracellular protein, DNA and the like;
the small molecular medicine can be at least one of rapamycin, ciprofloxacin, levofloxacin and pipecolic acid;
in the nanoparticle, the mass ratio of the temperature-responsive amphiphilic molecule to the non-temperature-sensitive amphiphilic molecule may be 1:10-10:1, and may specifically be 8:3;
the ratio of the total mass of the conjugated polymer to the non-temperature-sensitive amphiphilic molecule to the temperature-responsive amphiphilic molecule is 1:16-25, which can be 1:22;
the small molecular medicine accounts for 3-40% of the whole mass, and does not contain other carriers.
The phage is selected according to the bacterial species of the bacterial biofilm to be destroyed;
for example, for Pseudomonas aeruginosa, pseudomonas aeruginosa phage (number: SHBCC D24610; source: shanghai collection biotechnology center) was selected.
According to the invention, the outer layer of the nanoparticle is modified with phage, the bacterial biofilm is precisely positioned by the phage, the conjugated polymer converts light energy into heat energy, and the phase change of amphiphilic molecules is initiated to release internal hydrophobic drugs, so that the bacterial biofilm is precisely and controllably damaged in time and space.
In another aspect, the present invention provides a method of preparing the above nanoparticle.
The preparation method of the nano particles provided by the invention comprises the following steps:
1) Dissolving a conjugated polymer and a non-temperature-sensitive amphiphilic molecule in a first solvent to obtain a first solution;
2) Dissolving temperature-responsive amphiphilic molecules and small molecule drugs in a second solvent to obtain a second solution;
3) Mixing the first solution with the second solution, and performing ultrasonic dispersion for 10-50 minutes;
4) Adding the obtained mixed solution into water, and performing ultrasonic treatment for 5-20 minutes;
5) Stirring the obtained liquid in a dark place, removing the organic solvent, and performing ultrasonic dispersion for 10-50 minutes;
6) Adding a carboxyl activating reagent into the mixture, and stirring the mixture for reaction for 1 to 5 hours;
7) Adding phage into the obtained system, stirring at 4-20deg.C for 5-36 hr, ultrafiltering to remove unconnected phage, concentrating, and diluting according to the required concentration.
In the above method step 1), the first solvent may be tetrahydrofuran, DMSO, ethanol;
in the above method step 2), the second solvent may be dichloromethane, chloroform or tetrahydrofuran;
the mass ratio of the temperature responsive amphiphilic molecules in step 2) to the non-temperature sensitive amphiphilic molecules in step 1) may be 1:10 to 10:1, in particular may be 8:3;
the ratio of the total mass of the conjugated polymer to the non-temperature-sensitive amphiphilic molecule and the temperature-responsive amphiphilic molecule in step 1) is 1:16-25, which can be 1:22;
in the step 4), the volume ratio of the mixed solution to the water is 1:0.8-1:10;
in step 6), the carboxyl activating reagent may specifically be 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide,
the mass ratio of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, the N-hydroxysuccinimide and the non-temperature sensitive amphiphilic molecule can be as follows: 1-3:0.8-1.5:1;
in step 7), the phage is added as a phage suspension in which phage (1-3). Times.10) 4 Each ml may be 2.4X10 4 Individual/ml;
1-2 volumes of phage suspension were added to each 1 volume of the system.
The application of the nano particles in preparing reagents for destroying bacterial biomembrane or medicines for treating related diseases caused by bacterial biomembrane infection also belongs to the protection scope of the invention.
The invention also provides a method of disrupting a bacterial biofilm.
The method for destroying the bacterial biological membrane provided by the invention comprises the following steps:
adding the nano particles into a bacterial biofilm system to be treated, and incubating for 10-60 minutes;
after the nano particles are combined with the bacterial biomembrane, the near infrared laser is used for irradiation, so that the nano particles generate a photo-thermal conversion effect to release the hydrophobic drug, damage the bacterial biomembrane and kill bacteria.
The invention adopts a simple self-assembly method to obtain the novel nano-drug with high dispersion, controllable particle size, good stability and guaranteed safety. The invention has the advantages of simple and pollution-free process, low cost, high efficiency, easy realization of industrial production and wide application prospect.
The invention provides multifunctional near infrared light responsive nano particles, which can accurately reach a bacterial biomembrane infection part, respond to near infrared light and quickly heat up, change the structure of the nano particles to directionally release medicines, realize the damage to the biomembrane by the cooperation of multiple actions, and overcome the generation and aggravation of bacterial drug resistance.
Drawings
FIG. 1 is a schematic representation of the preparation of PAP-CPNs@Levo according to example 1 of the present invention.
FIG. 2 shows the ultraviolet absorption spectrum of PAP-CPNs@Levo obtained in example 1 of the present invention.
FIG. 3 shows the measurement of the size and morphology of PAP-CPNs@Levo prepared in example 1 of the present invention by Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM).
FIG. 4 shows the concentration of PAP-CPNs@Levo at various concentrations in example 1 of the present invention under NIR laser irradiation (806 nm,0.6W cm) -2 ) Is a temperature rise curve of (a).
FIG. 5 shows the selectivity analysis of PAP-CPNs@Levo against P.aeruginosa in example 2 of the present invention.
FIG. 6 is a plate test and quantitative analysis of the antibacterial activity of PAP-CPNs@Levo against P.aeruginosa in example 3 of the present invention.
FIG. 7 shows the results of the experiment (crystal violet staining image of biofilm and quantitative plot of biofilm disruption degree) of the PAP-CPNs@Levo disrupting bacterial biofilm in example 4 of the present invention.
FIG. 8 shows the results of the experiment (3D CLSM image of the biofilm disruption of PAP-CPNs@Levo) of the bacterial biofilm disruption of PAP-CPNs@Levo in example 4 of the present invention.
FIG. 9 shows the cytotoxicity of PAP-CPNs@Levo on L929 cells in example 5 of the present invention.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Conjugated polymer P1 employed in the following examples: PDPP is prepared by a method comprising the following steps:
synthesis of PDPP: : 215mg Ni (cod) 2 106mg bipyridine to200mg of the monomer 3, 6-bis (5-bromothiophen-2-yl) -2, 5-di (octyldodecyl) pyrrolo [3,4-c]Pyrrole-1, 4-dione was added to toluene solvent, and three freeze-pump-thaw cycles were performed under argon to remove oxygen, and then the reaction mixture was heated to 80 ℃ to react for 20 hours. Then, a mixed solution of methanol, acetone and dilute hydrochloric acid was added to the system, and the mixture was stirred at room temperature for 1 hour. Chloroform extraction was used, and excess EDTA was added to the organic layer and stirred for 5 hours. The product was extracted sequentially with methanol, n-hexane, chloroform, the chloroform solution was collected and poured into methanol for precipitation, and 61mg of a dark blue-violet solid product was obtained by filtration, with a yield of 37%.
The embodiment of the invention takes the conjugated polymer PDPP as an example, and illustrates the application of phage-guided near-infrared response conjugated polymer nanoparticles in the aspect of destroying biological membranes, thereby describing the patent of the invention. The conjugated polymer PDPP has an absorption peak at 700-900nm and a higher extinction coefficient, and the light-heat conversion efficiency at 808nm is up to about 50%, and is 1Wcm -2 The aqueous solution having a concentration of 15. Mu.g/mL under near infrared light irradiation may be warmed up to 70 ℃.
Example 1 preparation of nanoparticles
Disclosed herein is a nanoparticle, which is formed by compositing particles assembled by amphiphilic molecules, hydrophobic drugs and conjugated polymers with phage;
the hydrophobic drug is not particularly limited, and is preferably a hydrophobic small molecular drug having a molecular weight of 300 to 5000, more preferably a molecule having a good antibacterial effect, for example, a molecule such as rapamycin, ciprofloxacin, levofloxacin, pipecolic acid, etc.;
the conjugated polymer adopted by the invention is a conjugated polymer which is subjected to chemical structure regulation and control to ensure that the conjugated polymer has stronger absorption in a near infrared region and higher photo-thermal efficiency. Conjugated polymers have been widely paid attention to fields such as optics and electricity in recent years due to their advantages of adjustable energy level, strong optical stability, high light absorption efficiency, and the like.
The temperature-responsive amphiphilic molecule used in this example was dipalmitoyl phosphatidylcholine (DPPC), the phase transition temperature of which was about 38 ℃, and the non-temperature-sensitive amphiphilic molecule was distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG-COOH);
the hydrophobic drug used in this embodiment is levofloxacin, which effectively inhibits the activity of DNA helicase in bacteria, effectively inhibits the proliferation of bacteria, and can inhibit the synthesis of protein in bacteria, which causes the bacteria to fail to secrete and metabolize normally, and induces bacterial necrosis, thereby treating diseases associated with bacterial infection.
The specific operation method of the embodiment is as follows:
0.5mg PDPP,3mg DSPE-PEG 2000 -COOH dissolved in 1mL tetrahydrofuran; 8mg of DPPC,1.5mg of levofloxacin are dissolved in 200uL of dichloromethane and sonicated for 30 minutes; mixing the two materials together, and continuing ultrasonic treatment for 30 minutes;
adding the mixed solution into 1mL of ultra-pure water, and performing ultrasonic treatment for 10 minutes;
the suspension was protected from light, stirred for 5 hours while simultaneously aeration was carried out to remove tetrahydrofuran and methylene chloride, sonicated for 30 minutes, and then passed through a 0.22 μm filter;
6mg EDC and 3mg NHS are added into the solution, 2mL MES buffer is added, and the mixture is stirred for 5 hours in a dark place;
5mL of phage suspension (phage concentration 2.4X10) 4 individual/mL), 4 ℃ for 24 hours;
and (3) ultrafiltering and concentrating the filtrate to obtain concentrated nano particles, calibrating the concentration of the nano particles according to PDPP, and diluting according to experimental requirements for use.
FIG. 1 shows the preparation of PAP-CPNs@Levo.
FIG. 2 is an ultraviolet-visible absorption spectrum of PAP-CPNs@Levo.
FIG. 3 is a transmission electron microscope image and particle size characterization of the prepared PAP-CPNs@Levo.
PAP-CPNs@Levo was diluted to different concentrations and irradiated with 808nm near infrared light as an excitation light source.
FIG. 4 shows the change in temperature with irradiation time of PAP-CPNs@Levo under near infrared light.
Example 2 Selective Effect of nanoparticles on Pseudomonas aeruginosa
Preparing fluorescent nano particles with or without PAP connection guidance;
incubating the fluorescent nanoparticles with or without PAP connection guidance with candida albicans, escherichia coli, staphylococcus aureus and pseudomonas aeruginosa for 30 minutes respectively;
washing the mixed solution with sterile water and centrifuging at 6000rpm for 5 minutes;
the final pellet was re-selected in water and the nanoparticle binding to bacteria was observed under laser confocal.
As shown in figure 5, the selectivity of the nanoparticle with PAP connection to Pseudomonas aeruginosa is stronger by laser confocal, which shows that the method can strengthen the selectivity of the nanoparticle to bacteria.
EXAMPLE 3 antibacterial Activity study of nanoparticles against Pseudomonas aeruginosa
The antibacterial activity of PAP-CPNs@Levo was evaluated by plate counting. Taking Pseudomonas aeruginosa as an example in the experimental process, washing the cultured Pseudomonas aeruginosa three times by PBS buffer solution, and preparing OD 600 Bacterial suspension=1.0, 100 μl of nanoparticles was mixed with 100 μl of bacterial suspension after ten-fold dilution. Incubate in the dark for 30 minutes, then at 0.6W cm -2 For 5 minutes, the NIR-irradiated Pseudomonas aeruginosa suspension was diluted 10 with PBS buffer 4 20. Mu.L of the bacterial liquid was taken out and uniformly spread on the solid LB medium. Then cultured for 18 hours and the colony count was calculated. The survival fraction is the ratio of the number of pseudomonas aeruginosa colonies not treated with near infrared light to the number of pseudomonas aeruginosa colonies treated with near infrared light.
As can be seen from fig. 6, PAP has weak bactericidal ability, and in the absence of illumination, PAP-cpns@levo has poor bacterial killing ability; under illumination, PAP-CPNs@Levo shows strong antibacterial activity, and the bacterial survival rate is lower than 1% when the nanoparticle concentration is 15 mug/mL.
EXAMPLE 4 investigation of the damaging Effect of nanoparticles on Pseudomonas aeruginosa biofilm
The embodiment utilizes the nanometer particles to reach the bacterial biomembrane infection position in a directional way, and the small molecular medicine is controllably released under the irradiation of near infrared light, and the related diseases caused by bacterial biomembrane infection are treated by killing bacteria and damaging biomembranes under the combined action of light and heat and the medicine. Methods of disrupting biological membranes by the nanoparticles of this patent include, but are not limited to, diseases treated using sterilization by near infrared light irradiation.
The biomembrane is a bacterial population formed by the aggregation of bacterial cells and the adhesion polymerization of extracellular matrixes such as polysaccharide, protein, extracellular DNA and the like, and can prevent the diffusion and permeation of the antibacterial agent. Phages are natural bacterial viruses, specific for bacteria and their secreted depolymerases contribute to disrupting biofilm formation, so that introduction of phages into nanoparticle construction has a very effective effect in targeting nanoparticles and deeply disrupting biofilms. The phage selected in this example was specific to simple copper.
The phage delivers the nanoparticle to the bacterial biofilm infection site and breaks the external polymer barrier, opening the channel for the photothermal action of the subsequent conjugated polymer. The nanometer particles can release the loaded medicine under the control of near infrared light irradiation, and the biological film is destroyed by the synergy of light and heat and the medicine to kill bacteria.
In the embodiment, the in-vitro biological film is prepared, the bacteria of the biological film are marked by using a crystal violet and live dead bacteria fluorescent dye kit, and the killing effect of the nano particles on the bacteria in the biological film under the irradiation of near infrared light of 808nm is analyzed to verify the treatment effect on the infection of the biological film.
The specific operation method of the embodiment is as follows:
placing a glass sheet with the diameter of 9mm in a 48-hole plate, adding bacterial LB culture medium suspension, and culturing at 37 ℃ for 48 hours to obtain an in-vitro mature biological film;
removing the upper culture solution, adding PBS buffer solution and PAP-CPNs@Levo nano particles respectively, and incubating for 30 minutes at 37 ℃; the method comprises the steps of carrying out a first treatment on the surface of the
Irradiating the illumination experiment group for 8min under 808nm laser with the laser power of 0.6W;
removing the buffer solution and the nano particles, and respectively adding crystal violet dye and Styo9/PI fluorescent dye to dye the bacteria for 15min under dark condition;
and (5) observing the dyeing condition, and observing the dyed fluorescent dye by using a laser confocal microscope.
The damage result of PAP-CPNs@Levo nano particles to the biological film is shown in the accompanying drawings 7 and 8, and after 808nm laser irradiation, the nano particles can penetrate into the biological film and can damage 70.7% of the biological film. These data indicate that the composite nanoparticles produced are capable of responding to NIR light, regulating bacterial biofilm formation processes, and effectively disrupting biofilms.
Example 5 cytotoxicity of the prepared nanoparticles the biocompatibility of the composite nanoparticles was evaluated using L929 cell viability assay
L929 cells were first grown at 2X 10 5 The density of each/mL is mixed with the nanoparticle equal volume containing different nanoparticle concentrations (4 ℃), then the mixture is placed in a 96-well plate for culture, 100 mu L of each well is carried out, 6 groups of the wells are parallel, and 100 mu L of culture medium is added after gel formation. After incubation for 24 hours, 10. Mu.L of CCK-8 was added to each well, and absorbance at 450nm was recorded with a microplate reader after 2 hours. Cell relative viability (%) = absorbance measured in experimental group/absorbance measured in control group x 100%.
FIG. 9 shows that the concentration of the nano particles provided by the invention is 0-30 mug/mL, the cytotoxicity to L929 is low, and the cell survival rate reaches 90%, so that the composite nano particles have good biocompatibility.
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains.
Claims (9)
1. A nanoparticle is prepared from amphipathic molecule, conjugated polymer, small molecule medicine and phage;
the nano particles are in a spherical shape, and the particle size is 50-500 nm.
2. The nanoparticle according to claim 1, wherein: the amphiphilic molecules comprise temperature-responsive amphiphilic molecules and non-temperature-sensitive amphiphilic molecules;
the conjugated polymer is any one of the following polymers:
the small molecular medicine is a hydrophobic medicine, and the target point of the small molecular medicine is bacterial cell membrane or biological molecule outside the cell membrane.
3. Nanoparticle according to claim 1 or 2, characterized in that: in the nanoparticle, the mass ratio of the temperature-responsive amphiphilic molecules to the non-temperature-responsive amphiphilic molecules is 1:10-10:1;
the ratio of the total mass of the conjugated polymer to the non-temperature-sensitive amphiphilic molecule to the temperature-responsive amphiphilic molecule is 1:16-25;
the small molecular medicine accounts for 3-40% of the whole mass, and does not contain other carriers.
4. A method of preparing the nanoparticle of any one of claims 1-3, comprising the steps of: 1) Dissolving a conjugated polymer and a non-temperature-sensitive amphiphilic molecule in a first solvent to obtain a first solution;
2) Dissolving temperature-responsive amphiphilic molecules and small molecule drugs in a second solvent to obtain a second solution;
3) Mixing the first solution with the second solution, and performing ultrasonic dispersion for 10-50 minutes;
4) Adding the obtained mixed solution into water, and performing ultrasonic treatment for 5-20 minutes;
5) Stirring the obtained liquid in a dark place, removing the organic solvent, and performing ultrasonic dispersion for 10-50 minutes;
6) Adding a carboxyl activating reagent into the mixture, and stirring the mixture for 1 to 5 hours;
7) Adding bacteriophage into the obtained system, stirring at 4-20deg.C for 5-36 hr, ultrafiltering, and concentrating.
5. The use according to claim 4, characterized in that: in step 7), the phage is added as a phage suspension in which phage (1-3). Times.10) 4 And each ml.
6. Use of a nanoparticle according to any one of claims 1 to 3 in the preparation of a reagent for disrupting a bacterial biofilm.
7. Use of the nanoparticle of any one of claims 1-3 in the manufacture of a medicament for the treatment of a disease associated with a bacterial biofilm infection.
8. Use according to claim 6 or 7, characterized in that: the application is as follows:
adding the nanoparticle of any one of claims 1-3 to a bacterial biofilm system to be treated, incubating for 10-60 minutes;
after the nano particles are combined with the bacterial biomembrane, the near infrared laser is used for irradiation, so that the nano particles generate a photo-thermal conversion effect to release the hydrophobic drug, damage the bacterial biomembrane and kill bacteria.
9. A method of disrupting a bacterial biofilm comprising the steps of:
adding the nanoparticle of any one of claims 1-3 to a bacterial biofilm system to be treated, incubating for 10-60 minutes;
after the nano particles are combined with the bacterial biomembrane, the near infrared laser is used for irradiation, so that the nano particles generate a photo-thermal conversion effect to release the hydrophobic drug, damage the bacterial biomembrane and kill bacteria.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310142147.3A CN116159039A (en) | 2023-02-21 | 2023-02-21 | Near infrared light response conjugated polymer/phage composite nanoparticle for destroying biological film |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310142147.3A CN116159039A (en) | 2023-02-21 | 2023-02-21 | Near infrared light response conjugated polymer/phage composite nanoparticle for destroying biological film |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116159039A true CN116159039A (en) | 2023-05-26 |
Family
ID=86419710
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310142147.3A Pending CN116159039A (en) | 2023-02-21 | 2023-02-21 | Near infrared light response conjugated polymer/phage composite nanoparticle for destroying biological film |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116159039A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080057038A1 (en) * | 2005-03-08 | 2008-03-06 | Iftach Yacoby | Targeted drug-carrying bacteriophages |
| CN106132440A (en) * | 2014-02-07 | 2016-11-16 | 固定噬菌体有限公司 | The treatment that locally and systemically property antibacterial infects |
| CN108795883A (en) * | 2018-06-07 | 2018-11-13 | 江苏大学 | A method of improving Phage Infection bacterium efficiency |
| CN110693852A (en) * | 2019-11-18 | 2020-01-17 | 河北工业大学 | Conjugated polymer-based near-infrared light response photothermal effect nanoparticles and preparation and application thereof |
| CN115607513A (en) * | 2022-11-15 | 2023-01-17 | 河北工业大学 | Preparation method of siderophore biomimetic nanoparticles and application of siderophore biomimetic nanoparticles in antibacterial aspect |
-
2023
- 2023-02-21 CN CN202310142147.3A patent/CN116159039A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080057038A1 (en) * | 2005-03-08 | 2008-03-06 | Iftach Yacoby | Targeted drug-carrying bacteriophages |
| CN106132440A (en) * | 2014-02-07 | 2016-11-16 | 固定噬菌体有限公司 | The treatment that locally and systemically property antibacterial infects |
| CN108795883A (en) * | 2018-06-07 | 2018-11-13 | 江苏大学 | A method of improving Phage Infection bacterium efficiency |
| CN110693852A (en) * | 2019-11-18 | 2020-01-17 | 河北工业大学 | Conjugated polymer-based near-infrared light response photothermal effect nanoparticles and preparation and application thereof |
| CN115607513A (en) * | 2022-11-15 | 2023-01-17 | 河北工业大学 | Preparation method of siderophore biomimetic nanoparticles and application of siderophore biomimetic nanoparticles in antibacterial aspect |
Non-Patent Citations (1)
| Title |
|---|
| 李玲丽: "纳米材料的生物响应、生物合成及利用", 中国博士学位论文全文数据库 工程科技I辑, no. 01, 15 January 2018 (2018-01-15), pages 020 - 90 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Li et al. | Calixarene/pillararene-based supramolecular selective binding and molecular assembly | |
| Karimi et al. | Smart nanostructures for cargo delivery: uncaging and activating by light | |
| Zhou et al. | Cancer cell membrane camouflaged semi‐yolk@ spiky‐shell nanomotor for enhanced cell adhesion and synergistic therapy | |
| Sun et al. | Light-activated biodegradable covalent organic framework-integrated heterojunction for photodynamic, photothermal, and gaseous therapy of chronic wound infection | |
| Wang et al. | Photothermal-responsive conjugated polymer nanoparticles for the rapid and effective killing of bacteria | |
| Wang et al. | pH responsive superporogen combined with PDT based on poly Ce6 ionic liquid grafted on SiO2 for combating MRSA biofilm infection | |
| Gao et al. | Chlorin e6 (Ce6)-loaded supramolecular polypeptide micelles with enhanced photodynamic therapy effect against Pseudomonas aeruginosa | |
| Zhang et al. | Killing three birds with one stone: Near-infrared light triggered nitric oxide release for enhanced photodynamic and anti-inflammatory therapy in refractory keratitis | |
| CN103908429A (en) | Thermosensitive liposome and preparation method and application thereof | |
| Zhou et al. | Near-infrared light-triggered synergistic phototherapy for antimicrobial therapy | |
| Nakonechny et al. | Eradication of gram‐positive and gram‐negative bacteria by photosensitizers immobilized in polystyrene | |
| Wang et al. | Enhanced photodynamic inactivation for Gram-negative bacteria by branched polyethylenimine-containing nanoparticles under visible light irradiation | |
| Xie et al. | Photonic hydrogels for synergistic visual bacterial detection and on-site photothermal disinfection | |
| ElZorkany et al. | Photothermal versus photodynamic treatment for the inactivation of the bacteria Escherichia coli and Bacillus cereus: an in vitro study | |
| Garg et al. | Fluorescein–metal hybrid surfactant conjugates as a smart material for antimicrobial photodynamic therapy against Staphylococcus aureus | |
| CN110693852B (en) | Conjugated polymer-based near-infrared light response photothermal effect nanoparticles and preparation and application thereof | |
| Vallejo et al. | An insight into the role of non-porphyrinoid photosensitizers for skin wound healing | |
| Qi et al. | Development of local anesthetic drug delivery system by administration of organo-silica nanoformulations under ultrasound stimuli: in vitro and in vivo investigations | |
| Wang et al. | Bacteria-triggered radical anions amplifier of pillar [5] arene/perylene diimide nanosheets with highly selective antibacterial activity | |
| Yi et al. | Nanozyme‐based supramolecular self‐assembly as an artificial host defense system for treatment of bacterial infections | |
| Ren et al. | Stereocomplex Crystallized Nanomedical System for Enhanced Type I PDT and Synergistic Chemo-Phototherapy | |
| Ni et al. | COVID-19-inspired “artificial virus” to combat drug-resistant bacteria by membrane-intercalation-photothermal-photodynamic multistage effects | |
| CN116159039A (en) | Near infrared light response conjugated polymer/phage composite nanoparticle for destroying biological film | |
| CN115607513B (en) | Preparation method of siderophore bionic nano particles and application of siderophore bionic nano particles in antibacterial aspect | |
| Su et al. | Photothermal-driven disassembly of naphthalocyanine nano-photosensitizers for photothermal and photodynamic therapy |
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 |