CN111154015A - Porphyrin-terminated nano-grade fluorescent polyrotaxane as well as preparation method and application thereof - Google Patents
Porphyrin-terminated nano-grade fluorescent polyrotaxane as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN111154015A CN111154015A CN202010004770.9A CN202010004770A CN111154015A CN 111154015 A CN111154015 A CN 111154015A CN 202010004770 A CN202010004770 A CN 202010004770A CN 111154015 A CN111154015 A CN 111154015A
- Authority
- CN
- China
- Prior art keywords
- polyrotaxane
- porphyrin
- drug
- terminated
- fluorescent
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 239000003814 drug Substances 0.000 claims abstract description 63
- 239000003937 drug carrier Substances 0.000 claims abstract description 24
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 21
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims abstract description 20
- 150000004032 porphyrins Chemical class 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000007787 solid Substances 0.000 claims abstract description 14
- 238000005406 washing Methods 0.000 claims abstract description 11
- 238000006243 chemical reaction Methods 0.000 claims abstract description 10
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims abstract description 9
- 239000002244 precipitate Substances 0.000 claims abstract description 8
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 239000008367 deionised water Substances 0.000 claims abstract description 4
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 4
- 238000002156 mixing Methods 0.000 claims abstract description 3
- 239000000203 mixture Substances 0.000 claims abstract description 3
- 239000002904 solvent Substances 0.000 claims abstract description 3
- 229940079593 drug Drugs 0.000 claims description 52
- 239000012153 distilled water Substances 0.000 claims description 10
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 claims description 6
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 claims description 6
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 6
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 5
- 229960004316 cisplatin Drugs 0.000 claims description 5
- 238000000502 dialysis Methods 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 239000000700 radioactive tracer Substances 0.000 claims description 4
- 239000012265 solid product Substances 0.000 claims description 4
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 claims description 3
- 235000019260 propionic acid Nutrition 0.000 claims description 3
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 229940014800 succinic anhydride Drugs 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 abstract description 29
- 230000000694 effects Effects 0.000 abstract description 18
- 238000002474 experimental method Methods 0.000 abstract description 11
- 239000002246 antineoplastic agent Substances 0.000 abstract description 9
- 229940041181 antineoplastic drug Drugs 0.000 abstract description 9
- 231100000331 toxic Toxicity 0.000 abstract description 8
- 230000002588 toxic effect Effects 0.000 abstract description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 36
- 210000004027 cell Anatomy 0.000 description 29
- 239000000463 material Substances 0.000 description 12
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Substances [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 12
- 241000699670 Mus sp. Species 0.000 description 9
- 239000000243 solution Substances 0.000 description 8
- 229920000858 Cyclodextrin Polymers 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 6
- 239000002994 raw material Substances 0.000 description 6
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 6
- 239000002202 Polyethylene glycol Substances 0.000 description 5
- 230000000259 anti-tumor effect Effects 0.000 description 5
- 229920001223 polyethylene glycol Polymers 0.000 description 5
- 230000007547 defect Effects 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 239000002086 nanomaterial Substances 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 239000002105 nanoparticle Substances 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 231100000820 toxicity test Toxicity 0.000 description 3
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 238000001215 fluorescent labelling Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- UJKPHYRXOLRVJJ-MLSVHJFASA-N CC(O)C1=C(C)/C2=C/C3=N/C(=C\C4=C(CCC(O)=O)C(C)=C(N4)/C=C4\N=C(\C=C\1/N\2)C(C)=C4C(C)O)/C(CCC(O)=O)=C3C Chemical compound CC(O)C1=C(C)/C2=C/C3=N/C(=C\C4=C(CCC(O)=O)C(C)=C(N4)/C=C4\N=C(\C=C\1/N\2)C(C)=C4C(C)O)/C(CCC(O)=O)=C3C UJKPHYRXOLRVJJ-MLSVHJFASA-N 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- AOBORMOPSGHCAX-UHFFFAOYSA-N Tocophersolan Chemical compound OCCOC(=O)CCC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C AOBORMOPSGHCAX-UHFFFAOYSA-N 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 210000001099 axilla Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- -1 benzotriazole-1-yl-oxy tripyrrolidinyl phosphorus hexafluorophosphate Chemical compound 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000021523 carboxylation Effects 0.000 description 1
- 238000006473 carboxylation reaction Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000000214 effect on organisms Effects 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 150000003278 haem Chemical class 0.000 description 1
- 229960003569 hematoporphyrin Drugs 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000005918 in vitro anti-tumor Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0012—Cyclodextrin [CD], e.g. cycle with 6 units (alpha), with 7 units (beta) and with 8 units (gamma), large-ring cyclodextrin or cycloamylose with 9 units or more; Derivatives thereof
- C08B37/0015—Inclusion compounds, i.e. host-guest compounds, e.g. polyrotaxanes
-
- 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
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/243—Platinum; Compounds thereof
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
- A61K47/6951—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0036—Porphyrins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Polymers & Plastics (AREA)
- Materials Engineering (AREA)
- Inorganic Chemistry (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a preparation method of porphyrin-terminated nano-scale fluorescent polyrotaxane, which comprises the steps of mixing pseudorotaxane, porphyrin, PyBOP, DMAP and solvent anhydrous DMF, placing the mixture in a shaking table to react for 12-24h at the temperature of 2-8 ℃, adding methanol to separate out solids after the reaction is finished, washing the solids, dispersing the solids in DMSO, adding deionized water to separate out light pink precipitates, washing and freeze-drying the precipitates to obtain the porphyrin-terminated nano-scale fluorescent polyrotaxane. The invention also provides an antitumor drug prepared by using the nanoscale fluorescent polyrotaxane as a drug carrier. The experimental experiment shows that: the nano-scale fluorescent polyrotaxane can trace the medicine, improve the enrichment of the medicine in tumor parts, and reduce the toxic and side effects of the medicine on organisms, thereby improving the treatment effect of the medicine on the tumor.
Description
Technical Field
The invention belongs to the technical field of medicine preparation, and particularly relates to a porphyrin-terminated nanoscale fluorescent polyrotaxane as well as a preparation method and application thereof.
Background
The detection of tumors and the enrichment of drugs at the tumor sites are key to cancer treatment. Therefore, the preparation of the fluorescent nano drug-carrying system with the tracing function is imperative. At present, clinically used antitumor drugs have no targeting property, cannot be traced and have serious toxic and side effects in the treatment process of tumors. Nanomaterials have been the focus of research because of their enhanced penetration and retention effects in tumors. However, most of the currently studied nano-drug carriers have the defects of complex preparation process, high cost, poor biocompatibility of used materials, no tracing function of drugs and the like.
Based on the defects, the application discloses a fluorescent nano drug-loaded system with targeting and drug tracing functions. In the system, the prepared nano drug-loaded material has fluorescence due to the introduction of porphyrin, so that the fluorescent labeling of organic dye in later use is avoided, and the fluorescent drug-loaded material can be used as a tracer of a drug to detect the amount of the drug reaching the tumor part.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the porphyrin-terminated nanoscale fluorescent polyrotaxane which is a nano drug-carrying system prepared from porphyrin, cyclodextrin and polyethylene glycol as raw materials, can trace a drug, improve the enrichment of the drug at a tumor part, reduce the toxic and side effects of the drug on organisms and improve the treatment effect of the drug on tumors.
The invention also provides a preparation method and application of the porphyrin-terminated nano-fluorescent polyrotaxane
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of porphyrin-terminated nano-fluorescent polyrotaxane comprises the following steps: mixing pseudorotaxane (PPR), porphyrin, PyBOP (benzotriazole-1-yl-oxy tripyrrolidinyl phosphorus hexafluorophosphate), DMAP (4-dimethylaminopyridine) and solvent anhydrous DMF (dimethylformamide), placing the mixture in a shaking table to react for 12-24h at the temperature of 2-8 ℃, adding methanol to separate out solid after the reaction is finished, washing the solid, dispersing the solid in DMSO (dimethyl sulfoxide), adding deionized water to separate out light pink precipitate, washing and freeze-drying to obtain the porphyrin-terminated nano-grade fluorescent polyrotaxane PR.
Further, the molar ratio of the pseudorotaxane, the porphyrin, the PyBOP and the DMAP is 1: 10-12: 10-12: 1-3.
Further preferably, the porphyrin is prepared by the following steps: adding pyrrole and benzaldehyde into propionic acid according to the molar ratio of 1:1, stirring and heating to 130-150 ℃, carrying out reflux reaction for 30-60 min, and then cooling, carrying out suction filtration, washing and drying to obtain the compound.
The invention provides a porphyrin-terminated nano-fluorescent polyrotaxane PR prepared by the preparation method.
The invention provides application of the porphyrin-terminated nano-fluorescent polyrotaxane as a drug carrier or a drug tracer.
The invention also provides a drug-loaded nano system prepared by using the porphyrin-terminated nano-fluorescent polyrotaxane as a drug carrier, wherein the porphyrin-terminated nano-fluorescent polyrotaxane PR, succinic anhydride and DMAP are dissolved in DMSO and react for 12-24h at room temperature, then the reaction solution is dialyzed for 24-48h in distilled water, and the dialyzed solution is freeze-dried to obtain a light green solid product carboxylated polyrotaxane PR-COOH; and dissolving the carboxylated polyrotaxane and cisplatin in distilled water, reacting for 24-48h at room temperature in a dark place, dialyzing for 24-36 h in distilled water, and freeze-drying the dialyzed solution to obtain the drug-loaded nano system (namely the drug-loaded polyrotaxane, PR-COOH-Pt).
Further preferably, the dialysis bag with molecular weight cut-off of 3-5 kDa is selected.
The invention aims to improve the enrichment of the antitumor drug in a tumor part and reduce the toxic and side effects of the existing antitumor drug by tracing the distribution of the antitumor drug in a body, thereby overcoming the defects of complex preparation process, high cost, poor biocompatibility and the like of the existing drug carrier. Porphyrin has fluorescence, can form biocompatible hematoporphyrin in vivo and can be absorbed by organisms, and the biocompatibility of the host molecule cyclodextrin and the guest molecule polyethylene glycol for preparing the Polyrotaxane (PR) is good. Therefore, the nano drug-loaded system prepared by taking porphyrin, cyclodextrin and polyethylene glycol as raw materials can trace the drug, improve the enrichment of the drug in tumor parts, reduce the toxic and side effects of the drug on organisms and improve the treatment effect of the drug on tumors.
In the invention, the porphyrin is prepared from cheap raw materials by a simple synthesis method, and the host molecule cyclodextrin and the guest molecule polyethylene glycol used in the host-guest chemistry are not high in market price, so that the cost for preparing the drug-carrying material is low. From the biocompatibility point of view, cyclodextrin and polyethylene glycol are FDA approved materials with good biocompatibility, and porphyrin can be combined with iron in vivo to form heme to participate in the synthesis of protein. The invention researches the biological performance of the nano-drug carrier PR-COOH through in vitro and in vivo experiments. The distribution of PR-COOH in cells is researched through in vitro cell experiments, and the biocompatibility of PR-COOH and the in vitro anti-tumor effect of the loaded drug are researched through cytotoxicity experiments. Compared with the prior art, the invention has the following advantages and beneficial effects:
1) the raw materials for preparing PR-COOH are cheap and low in cost, good in biocompatibility and fluorescent, and are good drug tracers and drug carriers; the preparation method of the nano-drug carrier PR-COOH is simple and easy to operate, the cost for preparing a drug-carrying system is low, and the biocompatibility is good;
2) the nano-drug carrier PR-COOH has fluorescence, avoids organic dye fluorescence labeling and can trace drugs; the EPR effect of the tumor on the nano-drug carrier improves the drug concentration of the tumor part, has low toxic and side effects on organisms and enhances the tumor treatment effect.
Drawings
FIG. 1 shows (A) the hydrogen nuclear magnetic resonance spectra of polyrotaxane PR and pseudorotaxane PPR in example 1 and (B) the XRD spectra of cyclodextrin α -CD, pseudorotaxane PPR and polyrotaxane PR;
FIG. 2 is a transmission electron micrograph of carboxylated polyrotaxane PR-COOH (A) in example 1; (B) is a particle size distribution diagram of carboxylated polyrotaxane PR-COOH;
FIG. 3 is a confocal image of laser light generated after co-culturing the carboxylated polyrotaxane PR-COOH with CT26 cells and Hela cells at 37 ℃ in example 1;
FIG. 4 is a toxicity test of carboxylated polyrotaxane PR-COOH on (A) HL7702 cells (B) CT26 cells l (C) HeLa cells in example 1;
FIG. 5 is a toxicity test of cisplatin CDDP and the drug-loaded nanomaterial PR-COOH-Pt of example 1 on (A) CT26 cell (B) HeLa cell;
FIG. 6 (A) shows the change of tumor volume in tumor-bearing mice of different experimental groups; (B) change in body weight of tumor-bearing mice for different experimental groups; (C) the survival time of tumor-bearing mice of different experimental groups.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the following examples, but the scope of the present invention is not limited thereto.
The raw Materials used in the present invention can be purchased as ordinary commercial products, wherein the pseudorotaxane (PPR) can be prepared according to the prior art, such as Yi Zhang, Qiaoqiao Zhou, Shuxin Jia, Kunpeng Lin, guangfangFan, Jintao Yuan, Shuling Yu, and Jianhua Shi, Specific Modification with TPGS and Drug Loading of cyclic Modification polypeptides and the enhanced Activity Study in Vitro and in Vivo, Applied Materials & Interfaces, 2019, 11, 50, 46427 46436.
Example 1
A preparation method of porphyrin-terminated nano-fluorescent polyrotaxane comprises the following steps:
pseudorotaxane (3.0471 g, 0.1 mmol), porphyrin (629.24 mg, 1.0 mmol), PyBOP (520 mg, 1.0 mmol) and DMAP (122.17 g, 0.1 mmol) were added sequentially to a 50 mL round bottom, followed by 20 mL anhydrous DMF and allowed to react overnight (12 h) in a shaker at 4 deg.C, after the reaction was complete 20 mL methanol was added, whereupon a purple solid precipitated. Centrifugally collecting purple solid, washing the purple solid with methanol, dichloromethane and distilled water in sequence, ultrasonically dispersing the obtained purple solid in 10 mL of DMSO, standing the purple solid to fully disassemble the residual pseudorotaxane, then dropwise adding 80 mL of deionized water to obtain light pink precipitate, centrifugally collecting the light pink precipitate, washing the precipitate with methanol and water respectively, and freeze-drying the precipitate to obtain the porphyrin-terminated nano-grade fluorescent Polyrotaxane (PR) which is a light pink solid product.
The porphyrin is prepared by the following steps: pyrrole (6.84 mL, 0.098 mol) and benzaldehyde (10.00 mL, 0.098 mol) were added to 320 mL of propionic acid and heated to 140 ℃ with stirring for 30 min under reflux. Cooling overnight (12 h), suction filtering, washing with methanol for several times to obtain purple crystal, and drying in a vacuum drying oven to obtain porphyrin.
A drug-loaded nano system prepared by taking the porphyrin-terminated nano-fluorescent polyrotaxane as a nano load material through loading an anti-tumor drug is as follows:
1) porphyrin-terminated nanoscale fluorescent polyrotaxane PR (100 mg, 3.36X 10)-3mmol), succinic anhydride (54 mg, 0.545 mmol) and DMAP (66 mg, 0.545 mmol) were dissolved in DMSO (20 mL), the mixed solution was reacted at room temperature overnight for 12 hours, then the reaction solution was put into a 3.5 kDa dialysis bag and dialyzed in distilled water for 24 hours, and finally the dialyzed solution was freeze-dried to obtain a pale green solid product (carboxylated polyrotaxane PR-COOH), 134.5 mg;
2) carboxylated polyrotaxane PR-COOH (30 mg, 7.55X 10)-4mmol) and cisplatin (CDDP, 4.8 mg, 0.016 mmol) were dissolved in 3 mL of distilled water, reacted for 24h at room temperature in the dark, then the reaction solution was packed into a 3.5 kDa dialysis bag and dialyzed in distilled water for 24h, and finally the dialyzed solution was freeze-dried to obtain the product drug-loaded nanosystem (PR-COOH-Pt) (24.2 mg, 72%).
The structure and morphology of the product prepared in example 1 above were characterized by nmr, XRD and tem techniques as shown in fig. 1 and 2, respectively. The test result proves that: polyrotaxane (PR) is successfully synthesized and is in the shape of a spherical-like nano particle with the particle size of about 1.7 nm, so that the nano particle with the small particle size has better permeation and retention effects in tumors, and the medicine concentration of the tumor part is higher after the nano particle is loaded with the medicine, so that the anti-tumor effect is improved.
In order to further understand the drug loading performance of the polyrotaxane PR, the polyrotaxane PR-COOH is carboxylated to obtain carboxylated polyrotaxane PR-COOH (the aim is to facilitate drug loading), and then the drug cis-platinum CDDP is loaded, so that the drug-loaded nano system PR-COOH-Pt is obtained. When the drug selected is not cisplatin, the polyrotaxane PR can be directly loaded with the drug without carboxylation.
Application experiments
The end capping agent of PR-COOH is porphyrin with red fluorescence, so the PR-COOH prepared by the invention also has red fluorescence. The invention selects a human cervical cancer cell HeLa and a mouse colon cancer cell CT26 to detect the distribution of PR-COOH in the cell.
Cells were incubated in RPMI 1640 medium containing 10% FBS (fetal bovine serum), 1% penicillin (100U/mL) and streptomycin (0.1 mg/mL) for 24h at 37 ℃. The cells were then incubated with 0.25 mg/mL PR-COOH for 4h at 37 ℃. After 4h, cells were washed with PBS buffer (pH 7.4). The cells were then fixed on a cover glass with 4% paraformaldehyde for 10min and the nuclei stained with DAPI (4', 6-diamidino-2-phenylindole). The sample was observed by laser scanning confocal microscope and the results are shown in fig. 3. The results of fig. 3 show that: the drug carrier PR-COOH can be taken up by both cancer cells by endocytosis and distributed in the cytoplasm.
The cytotoxicity of the drug carrier PR-COOH and the anti-tumor drug PR-COOH-Pt loaded on the nano material is researched through an MTT experiment. Cells were seeded at a density of 7000 cells per well in 96-well plates and incubated at 37 ℃ for 24 h. Then, PR-COOH at various concentrations (12.5, 25, 50, 100 and 200. mu.g/mL) and PR-COOH-Pt, CDDP at various concentrations (5, 10, 20, 40, 80. mu.g/mL) were incubated with the cells at 37 ℃ for 48 h. Then, 50. mu.L of PBS buffer containing 5 mg/mL MTT was added to each well and the cells were incubated for an additional 4 h. MTT was then removed and DMSO was added to dissolve formazan crystals resulting from reduction of MTT by living cells. The absorbance was measured at 570 nm using a microplate reader. Untreated cells served as a control group. In this experiment, CT26 cells, HL7702 cells and HeLa cells were used to evaluate the cytotoxicity and antitumor activity of PR-COOH, PR-COOH-Pt and CDDP.
The test results are shown in fig. 4 and 5. The experimental results of fig. 4 show that: the nano-drug carrier PR-COOH prepared by the invention has no toxicity to normal cells and various tumor cells, and the material is proved to have better biocompatibility. The toxicity test result of the nano carrier PR-COOH-Pt loaded with the anti-tumor drug in the figure 5 shows that: the drug loaded on the nano drug carrier PR-COOH can be released and can keep the original drug effect, and the drug loaded on the nano drug carrier PR-COOH can be better taken up by tumor cells through the action of the nano drug carrier, so that the anti-tumor drug loaded on the nano material has better anti-tumor effect compared with a naked drug CDDP.
The biocompatibility and the anti-tumor effect of the nano-drug carrier PR-COOH are further researched through in vivo experiments. Murine hepatoma cells H22 (2X 10 per mouse) were inoculated subcutaneously6Individual cells) were inoculated into the axilla of Kunming mice for 6-8 weeks to establish a tumor model. When the tumor volume grows to about 90 mm3At the time, mice were randomly divided into four groups (8 per group): (i) PR-COOH (same concentration as the drug-loaded material used in group iii), (ii) naked drug CDDP (5 mg/kg), (iii) PR-COOH-Pt (CDDP concentration 5 mg/kg) and (iv) normal saline were negative controls. Mice were injected with 200 μ L of saline solution of the relevant sample in vivo through the tail vein on the first, fourth and seventh days. Tumor size and body weight were measured every other day until day fifteen. The number of mice survived was also observed until the end of the experiment. The results of the study are shown in FIG. 6. The experimental results of fig. 6 show that: compared with a naked drug CDDP, the drug loaded in the nano drug carrier PR-COOH can better inhibit the tumor growth, namely the drug CDDP loaded on the nano drug carrier PR-COOH has better anti-tumor effect. The body weight experiment research shows that: in the whole experiment process, the weight of mice of the material PR-COOH and the drug-carrying material PR-COOH-Pt group is not obviously reduced, the material PR-COOH is proved to have better biocompatibility, and the drug CDDP loaded on the nano drug carrier PR-COOH can reduce the toxic and side effects of the drug on organisms; the survival rate experimental study shows that: when the study reaches 45 days, all mice in the nude drug CDDP group die, and four mice in the drug-loaded PR-COOH-Pt group survive, which shows that the drug loaded on the nano drug carrier PR-COOH canThe survival time of the mouse is better prolonged.
To sum up, the following steps are carried out: the PR prepared by the invention has the advantages of cheap raw materials, low cost, good biocompatibility and fluorescence, and is a better drug tracer and a better drug carrier. After the medicine is loaded on PR-COOH, the EPR effect of the tumor on the nano medicine carrier PR-COOH improves the medicine concentration of the tumor part, the toxic and side effect of the medicine on organisms is low, and the tumor treatment effect is obviously enhanced.
Claims (7)
1. A preparation method of porphyrin-terminated nanoscale fluorescent polyrotaxane is characterized by mixing pseudorotaxane, porphyrin, PyBOP, DMAP and solvent anhydrous DMF, placing the mixture in a shaking table to react for 12-24 hours at the temperature of 2-8 ℃, adding methanol to separate out solids after the reaction is finished, washing the solids, dispersing the solids in DMSO, adding deionized water to separate out light pink precipitate, washing and freeze-drying to obtain the porphyrin-terminated nanoscale fluorescent polyrotaxane.
2. The method of claim 1, wherein the molar ratio of the pseudorotaxane, porphyrin, PyBOP and DMAP is 1: 10-12: 10-12: 1-3.
3. The method of preparing a porphyrin-terminated nanoscale fluorescent polyrotaxane according to claim 1, wherein the porphyrin is prepared by the following steps: adding pyrrole and benzaldehyde into propionic acid according to the molar ratio of 1:1, stirring and heating to 130-150 ℃, carrying out reflux reaction for 30-60 min, and then cooling, carrying out suction filtration, washing and drying to obtain the compound.
4. The porphyrin-terminated nanoscale fluorescent polyrotaxane prepared by the preparation method of any one of claims 1 to 3.
5. Use of the porphyrin-terminated nanoscale fluorescent polyrotaxane of claim 4 as a drug carrier or drug tracer.
6. A drug-loaded nano system prepared by using the porphyrin-terminated nanoscale fluorescent polyrotaxane as a drug carrier in claim 4 is characterized in that the porphyrin-terminated nanoscale fluorescent polyrotaxane, succinic anhydride and DMAP are dissolved in DMSO and react for 12-24h at room temperature, then the reaction solution is dialyzed for 24-48h in distilled water, and the dialyzed solution is freeze-dried to obtain a light green solid product carboxylated polyrotaxane; dissolving the carboxylated polyrotaxane and the cisplatin in distilled water, reacting for 24-48h at room temperature in a dark place, dialyzing for 24-36 h in the distilled water, and freeze-drying the dialyzed solution to obtain the compound.
7. The drug-loaded nanosystems of claim 6, wherein a dialysis bag with a molecular weight cut-off of 3-5 kDa is selected for dialysis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010004770.9A CN111154015A (en) | 2020-01-03 | 2020-01-03 | Porphyrin-terminated nano-grade fluorescent polyrotaxane as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010004770.9A CN111154015A (en) | 2020-01-03 | 2020-01-03 | Porphyrin-terminated nano-grade fluorescent polyrotaxane as well as preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111154015A true CN111154015A (en) | 2020-05-15 |
Family
ID=70560993
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010004770.9A Pending CN111154015A (en) | 2020-01-03 | 2020-01-03 | Porphyrin-terminated nano-grade fluorescent polyrotaxane as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111154015A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112121166A (en) * | 2020-10-16 | 2020-12-25 | 河南大学 | Specific porphyrin self-transporting nano carrier material and preparation method thereof |
| CN114504648A (en) * | 2022-02-17 | 2022-05-17 | 河南大学 | Rod-shaped hybrid nano material, medicine containing same and preparation method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2012025923A (en) * | 2010-07-28 | 2012-02-09 | Univ Of Tokyo | Method for synthesizing polyrotaxane and new polyrotaxane |
| CN103432582A (en) * | 2013-08-16 | 2013-12-11 | 中国医学科学院生物医学工程研究所 | Preparation method for RGD-targeted porphyrin polymer nanomicelle |
| CN104204035A (en) * | 2012-03-30 | 2014-12-10 | 宇部兴产株式会社 | Process for preparing polyrotaxane having blocking groups |
-
2020
- 2020-01-03 CN CN202010004770.9A patent/CN111154015A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2012025923A (en) * | 2010-07-28 | 2012-02-09 | Univ Of Tokyo | Method for synthesizing polyrotaxane and new polyrotaxane |
| CN104204035A (en) * | 2012-03-30 | 2014-12-10 | 宇部兴产株式会社 | Process for preparing polyrotaxane having blocking groups |
| CN103432582A (en) * | 2013-08-16 | 2013-12-11 | 中国医学科学院生物医学工程研究所 | Preparation method for RGD-targeted porphyrin polymer nanomicelle |
Non-Patent Citations (2)
| Title |
|---|
| VALDERREY, VIRGINIA 等: "Polyatomic Anion Assistance in the Assembly of [2]Pseudorotaxanes", 《JOURNAL OF THE AMERICAN CHEMICAL SOCIETY》 * |
| 张艺: "两种不同载药聚轮烷的合成及生物性能研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112121166A (en) * | 2020-10-16 | 2020-12-25 | 河南大学 | Specific porphyrin self-transporting nano carrier material and preparation method thereof |
| CN114504648A (en) * | 2022-02-17 | 2022-05-17 | 河南大学 | Rod-shaped hybrid nano material, medicine containing same and preparation method |
| CN114504648B (en) * | 2022-02-17 | 2022-10-11 | 河南大学 | Rod-shaped hybrid nano material, medicine containing same and preparation method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110882396B (en) | Preparation method and application of tumor microenvironment and redox stepwise responsive nano drug delivery system | |
| CN114806546B (en) | Organic framework material based on fluorescent molecules and preparation method and application thereof | |
| CN109678909B (en) | Preparation method and application of platinum derivative and platinum-containing nano particle | |
| CN102898543B (en) | Water-soluble carbon nanotube and application thereof | |
| CN111154015A (en) | Porphyrin-terminated nano-grade fluorescent polyrotaxane as well as preparation method and application thereof | |
| Liu et al. | Oxygen self-sufficient amphiphilic polypeptide nanoparticles encapsulating bodipy for potential near infrared imaging-guided photodynamic therapy at low energy | |
| CN112999352A (en) | RGD/PTX @ ZIF-90 drug delivery system and preparation method thereof | |
| CN112121166B (en) | Specific porphyrin self-transporting nano carrier material and preparation method thereof | |
| CN108888774B (en) | Tripterine-dendrimer conjugate as well as preparation method and application thereof | |
| CN113082206B (en) | Macromolecule nitric oxide donor modified up-conversion nanoparticle, preparation method and application | |
| CN107929261B (en) | pH-sensitive targeted nanoparticle preparation loaded with chemotherapeutic drugs and preparation method thereof | |
| CN108395543B (en) | Modified polyrotaxane, medicine-carrying micelle based on polyrotaxane and preparation method and application of medicine-carrying micelle | |
| CN114081947A (en) | Active tumor targeted controlled-release nano micelle and preparation method and application thereof | |
| CN110917349B (en) | Bowl-shaped ISP (internet service provider) composite functional nano particle as well as preparation method and application thereof | |
| CN109939081B (en) | F3 polypeptide targeted nano organometallic framework materials (nMOFs) and preparation method thereof | |
| CN110755379B (en) | Targeted drug delivery system capable of resisting drug-resistant tumors and preparation method thereof | |
| CN111298140A (en) | Reduction of the T of the response1/T2Switching type MRI contrast agent, preparation method and application thereof | |
| CN107243000B (en) | Drug-loaded hybrid nanoparticles and preparation method thereof | |
| CN113769115B (en) | Up-conversion nano diagnosis and treatment integrated platform probe and preparation method thereof | |
| CN106421812B (en) | Preparation method and application of self-assembled ferroferric oxide nanoparticles | |
| CN115887686A (en) | Anti-tumor nano prodrug released in response to ROS (reactive oxygen species), and preparation method and application thereof | |
| CN111687428B (en) | Amphiphilic polymer mediated gold nanoparticle controllable assembly and preparation and application thereof | |
| CN110627978B (en) | Brush-shaped polymer with cellulose nanocrystals as matrix and preparation method and application thereof | |
| CN107857788B (en) | Glycosylated BODIPY derivative, surface sugar modified nano micelle and preparation method and application thereof | |
| CN109675047B (en) | Method for carrying out liposome modification on compound with free hydroxyl |
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: 20200515 |