CN113044818A - Graphite oxide phase carbon nitride, anticancer drug, and preparation method and application thereof - Google Patents
Graphite oxide phase carbon nitride, anticancer drug, and preparation method and application thereof Download PDFInfo
- Publication number
- CN113044818A CN113044818A CN202110279315.4A CN202110279315A CN113044818A CN 113044818 A CN113044818 A CN 113044818A CN 202110279315 A CN202110279315 A CN 202110279315A CN 113044818 A CN113044818 A CN 113044818A
- Authority
- CN
- China
- Prior art keywords
- carbon nitride
- phase carbon
- graphite oxide
- tri
- oxide phase
- 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
- JMANVNJQNLATNU-UHFFFAOYSA-N oxalonitrile Chemical compound N#CC#N JMANVNJQNLATNU-UHFFFAOYSA-N 0.000 title claims abstract description 203
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 189
- 229910002804 graphite Inorganic materials 0.000 title claims abstract description 184
- 239000010439 graphite Substances 0.000 title claims abstract description 184
- 238000002360 preparation method Methods 0.000 title claims abstract description 60
- 239000002246 antineoplastic agent Substances 0.000 title claims abstract description 46
- 229940041181 antineoplastic drug Drugs 0.000 title claims abstract description 43
- QMKYBPDZANOJGF-UHFFFAOYSA-N benzene-1,3,5-tricarboxylic acid Chemical compound OC(=O)C1=CC(C(O)=O)=CC(C(O)=O)=C1 QMKYBPDZANOJGF-UHFFFAOYSA-N 0.000 claims abstract description 22
- MWWSFMDVAYGXBV-RUELKSSGSA-N Doxorubicin hydrochloride Chemical compound Cl.O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 MWWSFMDVAYGXBV-RUELKSSGSA-N 0.000 claims abstract description 19
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims abstract description 19
- 229960002918 doxorubicin hydrochloride Drugs 0.000 claims abstract description 19
- 229910017604 nitric acid Inorganic materials 0.000 claims abstract description 19
- 239000003814 drug Substances 0.000 claims abstract description 16
- 229960000789 guanidine hydrochloride Drugs 0.000 claims abstract description 9
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 230000035484 reaction time Effects 0.000 claims description 18
- 238000006243 chemical reaction Methods 0.000 claims description 14
- 238000005406 washing Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 239000008055 phosphate buffer solution Substances 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 238000010992 reflux Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 5
- 239000012265 solid product Substances 0.000 claims description 5
- 230000007935 neutral effect Effects 0.000 claims description 4
- 230000001093 anti-cancer Effects 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 3
- 239000004480 active ingredient Substances 0.000 claims 1
- JYEUMXHLPRZUAT-UHFFFAOYSA-N 1,2,3-triazine Chemical group C1=CN=NN=C1 JYEUMXHLPRZUAT-UHFFFAOYSA-N 0.000 abstract description 95
- 239000000463 material Substances 0.000 abstract description 56
- 210000004027 cell Anatomy 0.000 abstract description 54
- 201000011510 cancer Diseases 0.000 abstract description 16
- 229910052760 oxygen Inorganic materials 0.000 abstract description 15
- 206010028980 Neoplasm Diseases 0.000 abstract description 13
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 12
- 239000001301 oxygen Substances 0.000 abstract description 12
- 229940079593 drug Drugs 0.000 abstract description 11
- 125000000524 functional group Chemical group 0.000 abstract description 11
- 210000003855 cell nucleus Anatomy 0.000 abstract description 9
- 238000012986 modification Methods 0.000 abstract description 7
- 230000004048 modification Effects 0.000 abstract description 6
- 238000012544 monitoring process Methods 0.000 abstract description 5
- 238000010306 acid treatment Methods 0.000 abstract description 4
- 238000012984 biological imaging Methods 0.000 abstract description 4
- 238000012377 drug delivery Methods 0.000 abstract description 2
- 239000002243 precursor Substances 0.000 abstract description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 260
- 229960004679 doxorubicin Drugs 0.000 description 114
- 238000011068 loading method Methods 0.000 description 32
- 230000000694 effects Effects 0.000 description 27
- 230000005284 excitation Effects 0.000 description 26
- 238000001179 sorption measurement Methods 0.000 description 19
- 229940009456 adriamycin Drugs 0.000 description 16
- 230000001965 increasing effect Effects 0.000 description 16
- 239000006228 supernatant Substances 0.000 description 12
- 239000002609 medium Substances 0.000 description 10
- 238000012360 testing method Methods 0.000 description 9
- 238000004833 X-ray photoelectron spectroscopy Methods 0.000 description 8
- 238000000634 powder X-ray diffraction Methods 0.000 description 7
- 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 6
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 229910052799 carbon Inorganic materials 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 239000002105 nanoparticle Substances 0.000 description 5
- 238000000089 atomic force micrograph Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000000799 fluorescence microscopy Methods 0.000 description 4
- 230000003834 intracellular effect Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 208000032467 Aplastic anaemia Diseases 0.000 description 3
- 229930040373 Paraformaldehyde Natural products 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000002131 composite material Substances 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- 238000002073 fluorescence micrograph Methods 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 239000012737 fresh medium Substances 0.000 description 3
- 229910021389 graphene Inorganic materials 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 230000002147 killing effect Effects 0.000 description 3
- 229920002866 paraformaldehyde Polymers 0.000 description 3
- 230000004962 physiological condition Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 241000338702 Cupido minimus Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 102100021669 Stromal cell-derived factor 1 Human genes 0.000 description 2
- 101710088580 Stromal cell-derived factor 1 Proteins 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- -1 carbon nitrides Chemical class 0.000 description 2
- 239000003575 carbonaceous material Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000012202 endocytosis Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- LUYQYZLEHLTPBH-UHFFFAOYSA-N perfluorobutanesulfonyl fluoride Chemical compound FC(F)(F)C(F)(F)C(F)(F)C(F)(F)S(F)(=O)=O LUYQYZLEHLTPBH-UHFFFAOYSA-N 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 239000010453 quartz Substances 0.000 description 2
- 238000001878 scanning electron micrograph Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- 238000003892 spreading Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000009210 therapy by ultrasound Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- QCQCHGYLTSGIGX-GHXANHINSA-N 4-[[(3ar,5ar,5br,7ar,9s,11ar,11br,13as)-5a,5b,8,8,11a-pentamethyl-3a-[(5-methylpyridine-3-carbonyl)amino]-2-oxo-1-propan-2-yl-4,5,6,7,7a,9,10,11,11b,12,13,13a-dodecahydro-3h-cyclopenta[a]chrysen-9-yl]oxy]-2,2-dimethyl-4-oxobutanoic acid Chemical compound N([C@@]12CC[C@@]3(C)[C@]4(C)CC[C@H]5C(C)(C)[C@@H](OC(=O)CC(C)(C)C(O)=O)CC[C@]5(C)[C@H]4CC[C@@H]3C1=C(C(C2)=O)C(C)C)C(=O)C1=CN=CC(C)=C1 QCQCHGYLTSGIGX-GHXANHINSA-N 0.000 description 1
- 229910000838 Al alloy Inorganic materials 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 239000003972 antineoplastic antibiotic Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000013590 bulk material Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000030570 cellular localization Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- RKTYLMNFRDHKIL-UHFFFAOYSA-N copper;5,10,15,20-tetraphenylporphyrin-22,24-diide Chemical group [Cu+2].C1=CC(C(=C2C=CC([N-]2)=C(C=2C=CC=CC=2)C=2C=CC(N=2)=C(C=2C=CC=CC=2)C2=CC=C3[N-]2)C=2C=CC=CC=2)=NC1=C3C1=CC=CC=C1 RKTYLMNFRDHKIL-UHFFFAOYSA-N 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003837 high-temperature calcination Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- LELOWRISYMNNSU-UHFFFAOYSA-N hydrogen cyanide Chemical compound N#C LELOWRISYMNNSU-UHFFFAOYSA-N 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000010983 kinetics study Methods 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 150000004965 peroxy acids Chemical class 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 231100000683 possible toxicity Toxicity 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000009789 rate limiting process Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000007789 sealing 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
- 239000002356 single layer Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- JGVWCANSWKRBCS-UHFFFAOYSA-N tetramethylrhodamine thiocyanate Chemical compound [Cl-].C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=C(SC#N)C=C1C(O)=O JGVWCANSWKRBCS-UHFFFAOYSA-N 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- DKZXTOPFCDDGGX-UHFFFAOYSA-N tri-s-triazine Chemical compound C1=NC(N23)=NC=NC2=NC=NC3=N1 DKZXTOPFCDDGGX-UHFFFAOYSA-N 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 229920003169 water-soluble polymer Polymers 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B21/00—Nitrogen; Compounds thereof
- C01B21/06—Binary compounds of nitrogen with metals, with silicon, or with boron, or with carbon, i.e. nitrides; Compounds of nitrogen with more than one metal, silicon or boron
- C01B21/0605—Binary compounds of nitrogen with carbon
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- 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
- A61K47/51—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 non-active ingredient being a modifying agent
- A61K47/52—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 non-active ingredient being a modifying agent the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/70—Crystal-structural characteristics defined by measured X-ray, neutron or electron diffraction data
- C01P2002/72—Crystal-structural characteristics defined by measured X-ray, neutron or electron diffraction data by d-values or two theta-values, e.g. as X-ray diagram
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/80—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/80—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70
- C01P2002/82—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70 by IR- or Raman-data
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/80—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70
- C01P2002/85—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70 by XPS, EDX or EDAX data
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/01—Particle morphology depicted by an image
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/01—Particle morphology depicted by an image
- C01P2004/03—Particle morphology depicted by an image obtained by SEM
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/01—Particle morphology depicted by an image
- C01P2004/04—Particle morphology depicted by an image obtained by TEM, STEM, STM or AFM
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2006/00—Physical properties of inorganic compounds
- C01P2006/60—Optical properties, e.g. expressed in CIELAB-values
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Inorganic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a graphite oxide phase carbon nitride, an anticancer drug and a preparation method and application thereof, belonging to the technical field of biological medicine, wherein trimesic acid and guanidine hydrochloride are taken as precursors, after concentrated nitric acid treatment is adopted, oxygen-containing functional groups are successfully introduced, the graphite phase carbon nitride is changed into a porous network structure, then high-temperature and high-pressure modification treatment is carried out, the network structure is broken into a nano-granular structure, and finally the graphite oxide phase carbon nitride fluorescent material with a triazine structure is prepared; and the graphite oxide phase carbon nitride is further used as a carrier to adsorb doxorubicin hydrochloride to obtain the anticancer drug, which can be effectively absorbed by cancer cells and enriched in cells and cell nucleuses, and can realize the simultaneous implementation of material positioning and drug monitoring. The invention has wide application prospect in the fields of drug delivery, preparation of cancer treatment drugs, biological imaging and tracing.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a graphite oxide phase carbon nitride and anticancer drug as well as a preparation method and application thereof.
Background
Fluorescence microscopic imaging technology plays a critical role in the field of biomedicine at present, however, the fluorescence emission of the pure nitrogen-doped graphite-phase carbon nitride with the existing tris-s-triazine structure with better stability is blue fluorescence, and the fluorescence emission is basically consistent with the color of a nuclear stain, so that certain interference is caused to the tracing of materials, and the defects of large particle size, poor water dispersibility and poor biocompatibility exist, and the application of the fluorescence microscopic imaging technology in biomedicine is hindered.
Disclosure of Invention
In view of the above disadvantages or defects, the present invention provides a graphite oxide phase carbon nitride, an anticancer drug, and a preparation method and an application thereof, which can solve the disadvantages of the prior art that graphite phase carbon nitride has large particle size, poor water dispersibility and biocompatibility, and does not meet the application requirements in biomedicine.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a preparation method of graphite oxide phase carbon nitride, which comprises the following steps:
step (1): stirring and mixing guanidine hydrochloride and trimesic acid in a mass ratio of 3-7: 1 to obtain a solid, placing the solid in a reaction container, heating to 5-7 ℃ at normal pressure and normal temperature every minute until the temperature reaches 340-360 ℃, reacting for 3-5 hours, cooling to room temperature, taking out the solid product, grinding into powder, washing and drying to obtain graphite-phase carbon nitride;
step (2): placing the graphite-phase carbon nitride obtained in the step (1) into a reaction container, adding concentrated nitric acid, refluxing for 70-80 hours at the temperature of 80-95 ℃, then centrifugally washing to be neutral, and drying to obtain porous graphite-phase carbon nitride;
and (3): and (3) placing the porous graphite phase carbon nitride obtained in the step (2) into a reaction kettle, adding distilled water, and reacting at the temperature of 170-200 ℃ for 7-10 hours to obtain the graphite oxide phase carbon nitride.
Further, in the step (1), the mass ratio of guanidine hydrochloride to trimesic acid is 5:1, the heating rate is 5 ℃/min, the temperature of the heating termination is 350 ℃, the reaction time is 3.5 hours, and the drying temperature is 80 ℃.
Further, in the step (2), the reflux temperature is 80 ℃, the reflux time is 72 hours, and the drying temperature is 50 ℃.
Further, the mass fraction of the concentrated nitric acid in the step (2) is more than 68 percent.
Further, the reaction temperature in the step (3) was 180 ℃ and the reaction time was 8 hours.
The invention also provides the graphite oxide phase carbon nitride prepared by the preparation method.
The invention also provides an anticancer drug, which takes the graphite oxide phase carbon nitride as a carrier.
Furthermore, the active component of the anticancer drug is doxorubicin hydrochloride.
Furthermore, the planar thickness of the anti-cancer drug is 105-110 nm.
Further, the characteristic peak of the X-ray powder diffraction of the anticancer drug is 25.02 °.
The invention also provides a preparation method of the anticancer drug, which comprises the following steps:
step (1): respectively dissolving graphite oxide phase carbon nitride and doxorubicin hydrochloride in a phosphate buffer solution with the pH value of 2-11 to obtain a solution A and a solution B;
step (2): and (2) mixing the solution A and the solution B obtained in the step (1) under a dark condition, reacting for 5-24 hours on a shaking table, and then separating, purifying and washing to obtain the anti-cancer drug.
Further, the pH of the phosphate buffer solution in step (1) was 7.4.
Further, the mass concentration of the graphite oxide phase carbon nitride in the solution A in the step (1) is 10-100 mug/mL, and preferably 40 mug/mL.
Further, the mass concentration of the doxorubicin hydrochloride in the solution B in the step (1) is 5-400mg/L, preferably 50-150 mg/L.
Further, the reaction time on the shaker in step (2) is 10 to 24 hours, preferably 10 hours.
The invention also provides application of the graphite oxide phase carbon nitride in preparing an anti-cancer medicament.
The invention has the following advantages:
1. the invention provides a graphite oxide phase carbon nitride and a preparation method thereof, wherein trimesic acid and guanidine hydrochloride are taken as precursors, and the graphite phase carbon nitride with a tris-s-triazine structure is converted into the graphite phase carbon nitride with the triazine structure by introducing the trimesic acid; after the treatment of concentrated nitric acid, oxygen-containing functional groups are successfully introduced, the graphite phase carbon nitride is changed into a porous network structure, then the surface of the porous network structure is modified by a high-temperature and high-pressure method, the network structure is broken into a nano-particle structure, and finally the graphite oxide phase carbon nitride with a triazine structure is prepared; compared with the existing graphite oxide phase carbon nitride with a triazine structure and the existing graphite oxide phase carbon nitride with a triazine structure, the graphite oxide phase carbon nitride with a triazine structure obtained by the preparation method has excellent stability, water dispersibility and biocompatibility under aqueous solution and physiological conditions; meanwhile, the graphite oxide phase carbon nitride provided by the invention can emit green fluorescence under the irradiation of visible light, and the graphite oxide phase carbon nitride can be used as a biological material and has practical application value in the fields of biological imaging and tracing; the graphite oxide phase carbon nitride has the characteristics of high load rate, endocytosis by cells and excellent drug-loading performance, and can improve the treatment effect of the anti-cancer drug;
2. the invention provides an anticancer drug and a preparation method thereof, wherein the anticancer drug is composed of the graphite oxide phase carbon nitride as a carrier and doxorubicin hydrochloride adsorbed, and can be effectively absorbed by cancer cells and enriched in the cells and cell nuclei. Meanwhile, the graphite oxide phase carbon nitride and the doxorubicin hydrochloride are combined through electrostatic acting force, so that the subsequent release of the doxorubicin hydrochloride is facilitated, more released doxorubicin hydrochloride enters cells, the killing effect on cancer cells is facilitated, and the preparation method has a wide application prospect in preparation of drugs for treating malignant tumors or aplastic anemia; on the other hand, the fluorescence property of the graphite oxide phase carbon nitride is utilized, so that the material positioning and the drug monitoring can be simultaneously carried out.
Drawings
FIG. 1 shows a triazine structure of graphite phase carbon nitride (tri-C) according to the present invention3N4) And a triazine-structured graphite oxide phase carbon nitride (tri-HC)3N4) X-ray powder diffraction (XRD) pattern of (a);
FIGS. 2a to 2c are Scanning Electron Microscope (SEM) images of the triazine structure graphite phase carbon nitride, the triazine structure graphite phase carbon nitride treated with nitric acid, and the triazine structure graphite oxide phase carbon nitride, respectively;
FIG. 2d is an Atomic Force Microscope (AFM) image of a triazine structured graphite oxide phase carbon nitride of the present invention;
FIG. 3a is a Fourier transform infrared (FT-IR) spectrum of a triazine structured graphitic phase carbon nitride and a triazine structured graphite oxide phase carbon nitride of the present invention;
FIG. 3b is an X-ray photoelectron spectroscopy (XPS) plot of a triazine structured graphite phase carbon nitride and a triazine structured graphite oxide phase carbon nitride of the present invention;
FIGS. 4a and 4b are Scanning Electron Microscope (SEM) images of commercially available Doxorubicin (DOX) and the inventive doxorubicin-loaded triazine structure graphite oxide phase carbon nitride, respectively;
FIGS. 4c and 4d are Atomic Force Microscope (AFM) images and thickness analysis of commercially available Doxorubicin (DOX) and the inventive doxorubicin-loaded triazine structure graphite oxide phase carbon nitride;
FIG. 5a is a Fourier transform infrared (FT-IR) spectrum of commercially available doxorubicin with a graphite oxide phase carbon nitride of triazine structure of the invention and a graphite oxide phase carbon nitride of triazine structure loaded with doxorubicin;
FIG. 5b is an X-ray powder diffraction (XRD) pattern of commercially available doxorubicin with the graphite oxide phase carbon nitride of triazine structure and the doxorubicin-loaded graphite oxide phase carbon nitride of triazine structure of the present invention;
FIG. 6a is an X-ray photoelectron spectroscopy (XPS) plot of a triazine structure graphite oxide phase carbon nitride and a doxorubicin-loaded triazine structure graphite oxide phase carbon nitride of the present invention;
FIG. 6b is a zeta potential plot of commercially available doxorubicin with the graphite oxide phase carbonitride of triazine structure of the present invention and the graphite oxide phase carbonitride of triazine structure loaded with doxorubicin;
FIG. 7 is a graph showing the effect of pH on the doxorubicin-loading effect of the graphite oxide-phase carbon nitride of triazine structure according to the present invention;
FIGS. 8a and 8b are graphs showing the effect of the shaker reaction time on the doxorubicin loading effect of the triazine structure graphite oxide phase carbon nitride of the present invention and adsorption kinetics, respectively;
FIGS. 9a and 9b are graphs of the effect of the initial concentration of doxorubicin on the loading effect of the graphite oxide-phase carbon nitride of the triazine structure of this example and the adsorption isotherm, respectively;
FIG. 10 is a graph showing the effect of pH on the release of graphite oxide-phase carbon nitride from the triazine structure loaded with doxorubicin according to the present invention;
FIG. 11a is a photograph showing water solubility of a graphite oxide phase carbon nitride of a tris-s-triazine structure and a graphite oxide phase carbon nitride of a triazine structure according to the present invention;
FIG. 11b is a photograph of a fluorescent photograph at 365nm of a graphite oxide phase carbon nitride of a tris-s-triazine structure and a graphite oxide phase carbon nitride of a triazine structure according to the present invention;
FIG. 11c is a photograph of the inventive doxorubicin-loaded tris-s-triazine structured graphite oxide phase carbon nitride and doxorubicin-loaded triazine structured graphite oxide phase carbon nitride;
FIGS. 11d and 11e are photographs of a triazine structure graphite phase carbon nitride and a triazine structure graphite oxide phase carbon nitride of the present invention before and after standing for two weeks, respectively;
FIG. 12a is a fluorescence image at 365nm of a graphite oxide phase carbon nitride of a tris-s-triazine structure, a graphite oxide phase carbon nitride of a triazine structure, and a graphite phase carbon nitride of a triazine structure according to the present invention;
FIG. 12b is a graph showing fluorescence emission spectra of a triazine structure graphite oxide phase carbon nitride of the present invention under different excitation waves;
FIG. 13 is a graph showing the effect of the triazine-structured graphite oxide-phase carbon nitride on the cell activity of fibroblasts (L929) according to the present invention;
FIG. 14 is a flow cytogram of the penetration effect of different concentrations of the triazine structured graphite oxide phase carbon nitride of the present invention on Cal cancer cells;
FIG. 15 is a flow cytogram showing the penetration effect of graphite oxide phase carbon nitride of triazine structure of the present invention on Cal27 cancer cells at various times;
FIG. 16 is a graph of fluorescence images of a triazine structure graphite oxide phase carbon nitride in Cal cancer cells at different times;
FIGS. 17a to c are fluorescence spectra of the graphite oxide phase carbon nitride of triazine structure, commercially available doxorubicin hydrochloride, and the graphite oxide phase carbon nitride of triazine structure loaded with doxorubicin according to the present invention, respectively;
fig. 18 is an intracellular distribution fluorescence image of commercially available doxorubicin hydrochloride and the triazine-structured graphite oxide-phase carbon nitride of the present invention in Cal cancer cells.
Detailed Description
The following examples and experimental examples are further illustrative of the present invention and are in no way intended to limit the scope of the present invention. The present invention is further illustrated in detail below with reference to examples and experimental examples, but it should be understood by those skilled in the art that the present invention is not limited to these examples and experimental examples. Also, those skilled in the art can make improvements or modifications to the present invention based on the description of the present invention, but these are included in the scope of the present invention.
The term "triazine-structured graphite-phase carbon nitride" in the present specification means, unless otherwise specified, a triazine-structured graphite-phase carbon nitride obtained by introducing trimesic acid conversion into a tris-triazine-structured graphite-phase carbon nitride, and the abbreviation "tri-C" in the drawings and the following text3N4"means; the term "triazine-structured graphite oxide phase carbon nitride" in the present specification refers to a triazine-structured graphite oxide phase carbon nitride modified with concentrated nitric acid and high temperature and pressure, and the abbreviation "tri-HC" is used in the drawings and the following3N4"means; unless otherwise stated, in this specificationThe term "tris-s-triazine structured graphitic carbon nitride" refers to conventional tris-s-C graphitic carbon nitrides obtained by high temperature calcination of guanidine hydrochloride, the abbreviation "tri-s-C" in the drawings and the following3N4"means; the term "graphite oxide phase carbon nitride of a tris-s-triazine structure" in the present specification refers to graphite oxide phase carbon nitride of a tris-s-HC structure modified with concentrated nitric acid and high temperature and pressure, and the abbreviation "tri-s-HC" is used in the drawings and the following3N4"means; the term "doxorubicin" in the present description refers to doxorubicin hydrochloride, which is the hydrochloride salt of the anthracycline broad-spectrum antitumor antibiotic doxorubicin (doxorubicin), and is indicated in the drawings and in the following by the abbreviation "DOX"; the term "triazine structure-loaded graphite oxide phase carbon nitride" in the present specification refers to an anticancer drug, and includes a triazine structure-loaded graphite oxide phase carbon nitride using the aforementioned triazine structure-loaded graphite oxide phase carbon nitride as a drug carrier, and doxorubicin-loaded triazine structure-loaded graphite oxide phase carbon nitride, and the abbreviation "tri-HC" in the drawings and the following text3N4-DOX "represents; the term "abnormal proliferation of cells" in the present specification means that the growth, differentiation and apoptosis of cells deviate from the normal growth cycle of cells; the term "diseases associated with abnormal cell proliferation" as used herein refers to malignant tumors or aplastic anemia.
Example 1
Example 1 provides a graphite oxide phase carbon nitride (tri-HC)3N4) The preparation method specifically comprises the following steps:
step (1): stirring and mixing guanidine hydrochloride and trimesic acid in a mass ratio of 5:1, placing the obtained solid in an aluminum alloy crucible positioned in the central area of a quartz tube (with the inner diameter of 25mm and the length of 1000mm), heating at normal pressure and temperature for 5 ℃ per minute until 350 ℃, reacting for 3.5 hours, cooling the quartz tube to room temperature, taking out the solid product, grinding the solid product into powder, alternately cleaning ultrapure water and ethanol for several times, and drying at the temperature of 80 ℃ to obtain graphite-phase carbon nitride (tri-C)3N4)；
Step (2): placing 1g of graphite-phase carbon nitride obtained in the step (1) into a reaction container, adding 40mL of concentrated nitric acid, refluxing for 72 hours at the temperature of 80 ℃, then centrifugally washing to be neutral, and drying at the temperature of 50 ℃ to obtain porous graphite-phase carbon nitride;
and (3): and (3) placing 100mg of the porous graphite phase carbon nitride obtained in the step (2) into a reaction kettle, adding distilled water, and reacting at the temperature of 180 ℃ for 8 hours to obtain the graphite oxide phase carbon nitride.
Example 1 also provides the above-mentioned graphite oxide phase carbon nitride (tri-HC)3N4) The graphite phase carbon nitride (tri-C) prepared in the step (1) and the step (2) of the preparation method3N4)。
This example 1 further provides an anticancer drug and a preparation method thereof, which specifically includes the following steps:
step (1): performing ultrasonic treatment on the prepared graphite oxide phase carbon nitride with the triazine structure for 30min at 1000W by using a cell crushing ultrasonic instrument, centrifuging for 10min at 5000rpm, and dissolving 1mL of graphite oxide phase carbon nitride and commercially available adriamycin (DOX) in a phosphate buffer solution with the pH value of 7.4 to obtain a solution A and a solution B; wherein the mass concentration of the adriamycin (DOX) is 100 mg/L;
step (2): and (2) mixing the solution A and the solution B obtained in the step (1) under a dark condition, reacting for 10 hours on a shaking table, centrifuging for 10min at 10000rpm, separating and purifying, and washing by using distilled water until the supernatant does not show red to prepare the anti-cancer medicament.
Example 2
Example 2 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 2 provides an anticancer drug and the preparation method thereof, which differ from the example 1 only in that: the pH of the phosphate buffer solution was 2.0.
Example 3
Example 3 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 3 provides an anticancer drug and the preparation method thereof, which differ from example 1 only in that: the pH of the phosphate buffer solution was 4.0.
Example 4
Example 4 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 4 provides an anticancer drug and the preparation method thereof, which differ from example 1 only in that: the pH of the phosphate buffer solution was 9.0.
Example 5
Example 5 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 5 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the pH of the phosphate buffer solution was 11.0.
Example 6
Example 6 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 6 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the reaction time on the shaker was 10 minutes.
Example 7
Example 7 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 7 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the reaction time on the shaker was 20 minutes.
Example 8
Example 8 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 8 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the reaction time on the shaker was 30 minutes.
Example 9
Example 9 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 9 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the reaction time on a shaker was 2 hours.
Example 10
Example 10 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 10 provides an anticancer drug and the preparation method thereof, which is different from that of example 1 only in that: the reaction time on a shaker was 5 hours.
Example 11
Example 11 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 11 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the reaction time on a shaker was 24 hours.
Example 12
Example 12 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 12 provides an anticancer drug and the preparation method thereof, which differs from the example 1 only in that: the reaction time on a shaker was 36 hours.
Example 13
Example 13 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 13 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the reaction time on a shaker was 48 hours.
Example 14
Example 14 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 14 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the mass concentration of the adriamycin (DOX) is 5 mg/L.
Example 15
Example 15 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 15 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the mass concentration of the adriamycin (DOX) is 10 mg/L.
Example 16
Example 16 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 16 provides an anticancer drug and the preparation method thereof, which differs from the example 1 only in that: the mass concentration of the adriamycin (DOX) is 50 mg/L.
Example 17
Example 17 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 17 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the mass concentration of the adriamycin (DOX) is 150 mg/L.
Example 18
This example 18 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 18 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the mass concentration of the adriamycin (DOX) is 200 mg/L.
Example 19
Example 19 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And the preparation method thereof is the same as that of example 1.
This example 19 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the mass concentration of the adriamycin (DOX) is 300 mg/L.
Example 20
This example 20 provides a graphite oxide phase carbon nitride (tri-HC)3N4) And method of preparation and example 1The same is true.
This example 20 provides an anticancer drug and the preparation method thereof, which differs from example 1 only in that: the mass concentration of the adriamycin (DOX) is 400 mg/L.
Comparative example 1
The comparative example 1 provides a graphite oxide phase carbon nitride with a tris-triazine structure and a preparation method thereof, and specifically comprises the following steps:
the method comprises the following steps: the guanidine hydrochloride is directly calcined for 3.5 hours at the temperature of 450 ℃ to obtain the common Tri-s-triazine-structured graphite-phase carbon nitride (Tri-s-triazine-based g-C)3N4，tri-s-C3N4)。
Step two: 1g of graphite-phase carbon nitride with a tris-s-triazine structure is refluxed for 72 hours at 80 ℃ by using 40mL of concentrated nitric acid, the obtained porous graphite-phase carbon nitride material with the tris-s-triazine structure is washed to be neutral by a centrifugal washing method, and is dried at 50 ℃; finally, 100mg of the two materials are respectively added into 30mL of distilled water and put into a high-temperature reaction kettle to react for 8h at 180 ℃ to obtain the graphite oxide phase carbon nitride (tri-s-HC) with the tris-s-triazine structure3N4). Then ultrasonic treatment is carried out for 30min under 1000W by using a cell crushing ultrasonic instrument, and then centrifugation is carried out for 10min at 5000rpm, and supernatant is taken.
Experimental example 1
Experimental example 1 graphite oxide phase carbon nitride (tri-HC) obtained in example 13N4) Characterization was performed and the results are shown in FIGS. 1-3. Wherein FIG. 1 shows a triazine structure graphite phase carbon nitride (tri-C) obtained in example 13N4) And a triazine-structured graphite oxide phase carbon nitride (tri-HC)3N4) And tri-s-HC obtained in comparative example 13N4XRD pattern of (a); in FIG. 2, 2a and 2b are respectively a triazine structure graphite phase carbon nitride (tri-C) before and after the nitric acid treatment in example 13N4) FIG. 2c is a Scanning Electron Microscope (SEM) image of the triazine-structured graphite oxide phase carbon nitride (tri-HC) of example 13N4) FIG. 2d shows a triazine-structured graphite oxide phase carbon nitride (tri-HC) of example 13N4) An Atomic Force Microscope (AFM) image of (a); in FIG. 3FIG. 3a is the triazine structured graphite phase carbon nitride (tri-C) of example 13N4) And a triazine-structured graphite oxide phase carbon nitride (tri-HC)3N4) Fourier Transform Infrared (FTIR) spectrum of (A), FIG. 3b is a graph of a triazine-structured graphite-phase carbon nitride (tri-C) of example 13N4) And a triazine-structured graphite oxide phase carbon nitride (tri-HC)3N4) X-ray photoelectron spectroscopy (XPS) graph.
As shown in FIG. 1, the triazine structure tri-C3N4Medium 2 θ presents two characteristic peaks at approximately 16.09 ° and 26.19 °; carrying out surface modification treatment on the triazine structure graphite oxide phase carbon nitride (tri-HC) by using concentrated nitric acid and a high-temperature high-pressure method3N4) Peak at 27.19 ℃ is more tri-C3N4The peak at 26.19 degrees in the material is red-shifted and the peak shape is narrowed, indicating that the oxygen-containing functional group replaces part of-NH in the material2And the modified material has a better crystal structure. Furthermore, tri-HC3N4Small peak at 15.82 ℃ in comparison with tri-C3N4The small peak at 16.09 deg. in the (b) is blue-shifted due to the addition of the oxygen-containing functional group, increasing the spacing between the material layers, causing a blue shift in the peak position, further indicating the successful introduction of the oxygen-containing functional group.
As shown in FIG. 2a, tri-C without nitric acid treatment3N4Is a bulk material; after nitric acid treatment, the material becomes a porous fluffy structure (fig. 2 b); when the autoclave is used for high-temperature high-pressure treatment, the porous structure is thermally cut, and most of the porous structure is changed into nano particles (figure 2 c); the morphology indicates that the addition of oxygen-containing functional groups results in tri-C3N4Changing into porous network structure, and after high heat treatment, the network structure is broken into nano granular structure, which is also the modified tri-HC3N4The reason why the material solubility becomes good; FIG. 2d shows tri-HC3N4Nanoparticles around 50nm in size were present, essentially consistent with the SEM images. Review of morphology changes indicates that bulk tri-C3N4After concentrated nitric acid and high-temperature and high-pressure treatment, the nano-particles with the particle size of about 50nm are changed, which is helpful forThe water solubility is improved.
As shown in fig. 3a, the triazine structure graphite phase carbon nitride g-C before and after modification by concentrated nitric acid and high temperature and high pressure method3N4Three characteristic peaks are clearly observed in the material, and are respectively 3140cm-1、3280cm-1And 811cm-1Respectively represent stretching vibration and bending vibration of N-H in the triazine unit; tri-HC after modification treatment3N4Material, 1600cm-1The content of the oxygen-containing functional groups at the left and right parts is increased compared with that before treatment, and the characteristic peak of the triazine structure still obviously exists, which shows that after the treatment of concentrated nitric acid and high temperature and high pressure, the oxygen-containing functional groups are successfully introduced on the premise of keeping the triazine structure, thereby enhancing the water solubility of the material. As shown in FIG. 3b, tri-C3N4And tri-HC3N4C, N, O appears in the three elements, and the modified tri-HC3N4The content of O in the solution is more than tri-C3N4A significant increase indicates the successful introduction of oxygen-containing functional groups. In conclusion, the treatment of concentrated nitric acid and high temperature and high pressure does not change tri-C3N4And also can successfully introduce oxygen-containing functional groups, thereby improving the water solubility of the water-soluble polymer.
Experimental example 2
Experimental example 2 Tri-HC obtained in example 13N4DOX was characterized and the results are shown in FIGS. 4-6. Wherein FIG. 4a and FIG. 4b are respectively pure doxorubicin hydrochloride (DOX) and tri-HC obtained in example 13N4Scanning Electron Microscope (SEM) image of DOX, FIGS. 4c and 4d respectively of commercially pure doxorubicin hydrochloride (DOX) and tri-HC from example 13N4Atomic Force Microscope (AFM) image of DOX, FIG. 4e shows commercially pure doxorubicin hydrochloride (DOX) and tri-HC from example 13N4-AFM thickness analysis table for DOX; FIG. 5a and FIG. 5b in FIG. 5 are graphs of a triazine structure-loaded graphite oxide phase carbon nitride (tri-HC) before and after DOX loading3N4) Fourier Transform Infrared (FTIR) and X-ray powder diffraction (XRD) patterns of; FIGS. 6a and 6b of FIG. 6An X-ray photoelectron spectroscopy (XPS) diagram and a Zeta potential diagram of the triazine structure graphite phase carbon nitride before and after loading doxorubicin hydrochloride (DOX).
FIG. 4a shows a SEM image of a commercially available DOX showing a regular tetragonal structure when tri-HC3N4After loading with DOX (FIG. 4b), it is evident that DOX was spread over tri-HC3N4The surface layer of the nano-particles is graphite oxide phase carbon nitride (tri-HC) adsorbed on triazine structure by physical adsorption or electrostatic action3N4) A surface; as shown in FIG. 4c, pure DOX shows a regular structure with a size of about 100nm and a thickness of about 90nm, and when both loads are changed into tri-HC3N4DOX, shown by FIGS. 4d and 4e, of increased particle size compared to the first two, with a significant increase in thickness around 130nm, approximately 110nm, with visual indication of increased particle size and increased thickness of the loaded material particles, indicating tri-HC3N4Successful preparation of DOX.
As shown in FIG. 5a, tri-HC3N4After loading of the material and DOX, in tri-HC3N4Middle of-DOX Tri-HC3N4The material is 1265cm-1A new peak appears at the position, which represents the stretching vibration peak of the N-H bond at the outer ring of the benzene ring and is just matched with 1263cm in pure DOX-1The characteristic peaks are basically consistent, and the DOX and the tri-HC are indicated3N4Successful loading of the material; as shown in FIG. 5b, DOX has a characteristic peak at 25.02 deg., when compared to tri-HC3N4After loading, tri-HC3N4Substantially disappears, tri-HC3N4The characteristic diffraction peaks of DOX are substantially identical to those of DOX due to the large amount of DOX and tri-HC3N4Thereby showing a characteristic diffraction peak of DOX, and tri-HC3N4Almost no diffraction peak of (a) was observed.
As shown in FIG. 6a, after loading DOX, tri-HC3N4The increase of the proportions of C and O in DOX indicates that the introduction of C and O elements and the decrease of the proportion of N element in DOX are due to the fact that the content of N in DOX is lower and is far higher than that of C, O, so that the content ratio of N element is higherThe examples drop and also indirectly indicate successful loading of DOX. As shown in FIG. 6b, tri-HC3N4With a negative charge of-32.46 mV, indicating a modified tri-HC3N4The stability is better; DOX has a positive charge of 6.254mV, when loaded, tri-HC3N4-DOX showed a negative charge of-2.082 mV, indicating tri-HC3N4And DOX are combined through electrostatic acting force, so that subsequent release of DOX is facilitated.
Experimental example 3
This experimental example 3 examined the DOX loading effect of the anti-cancer drugs prepared at different pH values. The supernatant washed with distilled water to be non-reddish in step (2) of examples 1 to 5 was measured for the concentration of doxorubicin in the supernatant by ultra-high pressure liquid chromatography (UPLC), and the test results are shown in fig. 7. And calculating a load efficiency (E) and a load capacity (q) according to equation 1 and equation 2e)：
Load efficiency (%) - (W)Adding into-WFree form)/WAdding intoX 100% - -equation 1
(W) load capacity (mg/g)Adding into-WFree form)/WMaterialX 100% - -equation 2
Wherein, WAdding intoIs the initial concentration of doxorubicin, WFree formIs the concentration of adriamycin in the supernatant, WMaterialThe addition amount of the graphite oxide phase carbon nitride with a triazine structure.
As shown in fig. 7, neither peracid (pH 2.0) nor overbased (pH 11.0) conditions favor doxorubicin loading; and at the pH value of 7.4, the loading efficiency and the loading capacity of the graphene oxide on the adriamycin are highest.
This experimental example 3 examined the DOX loading effect of the anticancer drugs prepared at different shaker reaction times. The supernatant washed to non-reddish color by distilled water in step (2) of example 1 and examples 6 to 13 was measured for doxorubicin concentration in the supernatant by ultra-high pressure liquid chromatography (UPLC), and the loading efficiency (E) and loading capacity (q) were calculated according to the above-mentioned formula 1 and formula 2e) The test results are shown in FIG. 8, wherein FIGS. 8a and 8b are the table reaction time versus oxygen for triazine structures of the present inventionInfluence of adriamycin loading effect of graphite-phase carbon nitride and adsorption kinetics diagram.
As shown in fig. 8a, the adsorption efficiency and the adsorption capacity of the graphite oxide phase carbon nitride with the triazine structure to the adriamycin are increased along with the increase of the reaction time; wherein, tri-HC3N4The load efficiency for DOX rapidly increases within the first 10h, and when the load efficiency reaches 92% at 10h, the load efficiency is in a basically stable state along with the time, which indicates that the load efficiency of tri-HC3N4The load to DOX is rapid and the load efficiency is high. Therefore, the optimal reaction time of the triazine structure graphite oxide phase carbon nitride supported adriamycin in the invention is 10 h.
In addition, as shown in fig. 8b, based on the adsorption kinetics studies of the typical quasi-first order kinetic model (formula 3) and the quasi-second order kinetic model (formula 4), it is found that the fitting correlation of the quasi-second order kinetic model of graphene oxide of the present example is 0.9999, which is much higher than the fitting correlation of the quasi-first order kinetic model of 0.5316, indicating that tri-HC is obtained3N4The load behavior of DOX is more consistent with a quasi-second-order kinetic model, and the tri-HC is explained3N4The load behavior on DOX is a rate limiting process. Meanwhile, the experimental adsorption capacity (qexp) and the calculated adsorption capacity (qcal) in table 1 are very similar, further demonstrating tri-HC3N4The load behavior to DOX belongs to a quasi-second order kinetic model. The results show that tri-HC3N4The loading of DOX comprises the neutralization of DOX in a solution system with tri-HC3N4Diffusion at the surface, and diffusion within the material, such that tri-HC3N4The load on DOX is more sufficient.
TABLE 1 adsorption kinetics results
This experimental example 3 examined the DOX loading effect in the anti-cancer drugs prepared at different initial concentrations of doxorubicin. The supernatant washed to non-reddish color by distilled water in step (2) of example 1 and examples 14 to 20 was measured for the concentration of doxorubicin in the supernatant by ultra-high pressure liquid chromatography (UPLC), and the loading efficiency (E) and the loading capacity (q) were calculated according to the above-mentioned formula 1 and formula 2e) The test results are shown in fig. 9, in which fig. 9a and 9b are graphs of the effect of the initial concentration of doxorubicin on the loading effect of the graphite oxide-phase carbon nitride of the triazine structure of the present example and the adsorption isotherm, respectively.
As shown in fig. 9a, the loading efficiency and the loading capacity of the graphite oxide phase carbon nitride having a triazine structure according to the present invention gradually decreased and increased with the increase in the doxorubicin concentration. When the concentration of DOX is in the range of 100mg/L, the load efficiency is rapidly increased to 88.35%; when the concentration continued to increase, it appeared that tri-HC3N4The reduction in DOX loading efficiency indicates that at this concentration, the triazine-structured graphite oxide phase carbon nitride requires more active sites to accommodate the excess doxorubicin. Therefore, the equal volume of the graphite oxide phase carbon nitride composite 100mg/L adriamycin with the triazine structure of 0.5mg/mL is a better mixing ratio, and higher adsorption efficiency can be achieved.
Further, as shown in FIG. 9b and Table 2, it was found that the triazine structure graphite oxide phase carbon nitride of the present example can conform to the typical Langmuir adsorption isotherm model (equation 5) and Freundlich adsorption isotherm model (equation 6) based on adsorption isotherm studies2Both 1.000), which shows that both single-layer adsorption and multi-phase adsorption occur on the surface of graphene oxide, and the theoretical maximum adsorption capacity of the graphite oxide phase carbon nitride with the triazine structure to adriamycin can reach 2306 mg/g.
TABLE 2 adsorption isotherm results
Experimental example 4
This example 4 examined different pH and time vs. tri-HC3N4The effect of the release effect of DOX. The tri-HC obtained in example 1 was taken3N4DOX (4mL) was dissolved in 4mL PBS buffer solutions of different pH including pH 7.4 (normal physiological pH), pH 6.8 (pH in cancer tissue) and pH 5.0 (pH in cancer cell cytoplasm), after a certain time interval, 2mL of supernatants were removed, 2mL of fresh PBS solution was added, and so on to the set time points (20h, 40h, 60h, 80h, 100h and 120 h). Finally, the absorbance of the supernatant was measured using a fluorescence gradiometer, and the measurement results are shown in fig. 10. Calculating tri-HC in a certain time according to the accumulative drug release formula (formula 7)3N4Efficiency of release of DOX nanomedicine DOX (E,%):
E＝[CnV0+(C1+C2+…+Cn-1)V]/Wloadedx 100% - -equation 7
Wherein, Cn(mg/mL) is the concentration detected by taking out the supernatant for the nth time; v0(mL) volume of release medium; v (mL) is the volume of each sample; wloaded(mg) is the amount of DOX loaded on the material before release.
As shown in FIG. 10, tri-HC3N4DOX exhibits pH-dependent release, has certain persistence and shows certain slow-release effect. In addition to this, the present invention is,the highest release efficiency was achieved under acidic conditions, especially 5.0, up to 60.23%, with a release efficiency of 49.96% at pH 6.8, whereas the release efficiency was only 32.12% at 120h under normal physiological conditions 7.4, indicating a doxorubicin-loaded triazine-structured graphite oxide-phase carbon nitride (tri-HC)3N4DOX) is associated with the concentration of hydrogen ions, and doxorubicin is released more efficiently in the environment of the cancerous tissue.
Experimental example 5
In this example 5, the water dispersibility, fluorescence, stability and drug-loading performance of the graphite oxide phase carbon nitride having a triazine structure were examined. With tri-C obtained in example 13N4And tri-HC3N4And tri-s-HC obtained in comparative example 13N4For the purpose of examination, the test results are shown in FIG. 11, in which FIG. 11a is a photograph showing the water solubility of the graphite oxide phase carbon nitride of tris-s-triazine structure and the graphite oxide phase carbon nitride of triazine structure according to the present invention; FIG. 11b is a photograph of a fluorescent photograph at 365nm of a graphite oxide phase carbon nitride of a tris-s-triazine structure and a graphite oxide phase carbon nitride of a triazine structure according to the present invention; FIG. 11c is a photograph of the inventive doxorubicin-loaded tris-s-triazine structured graphite oxide phase carbon nitride and doxorubicin-loaded triazine structured graphite oxide phase carbon nitride;
fig. 11d and 11e are photographs of the triazine structure graphite phase carbon nitride and the triazine structure graphite oxide phase carbon nitride of the present invention before and after standing for two weeks, respectively.
As shown in FIG. 11a, a triazine-structured graphite oxide phase carbon nitride (tri-HC)3N4) The material shows better water dispersibility, namely the graphite oxide phase carbon nitride (tri-s-HC) with a tris-s-triazine structure3N4) The material still showed significant settling. As shown in FIG. 11b, tri-HC was irradiated under 365nm UV light3N4The material shows strong green fluorescence, and the tri-s-HC at the same concentration3N4The material exhibits very weak blue fluorescence; FIGS. 11d and 11e are respectively tri-C3N4And tri-HC3N4Standing the solution for two weeks to obtain images; as shown in FIG. 11dAnd 11e, the triazine-structured graphite oxide phase carbon nitride exhibits good water stability, and after standing for two weeks, tri-C3N4Substantially sink, and tri-HC3N4The material still shows good water dispersibility; as shown in FIG. 11c, the color change of the material before and after loading with doxorubicin clearly shows the tri-HC3N4The material shows better drug-loading performance.
This experimental example 5 also examined the optical properties of the graphite oxide phase carbon nitride of triazine structure. With tri-C obtained in example 13N4And tri-HC3N4And tri-s-C obtained in comparative example 13N4And tri-s-HC3N4For the object of examination, the test results are shown in fig. 12, in which fig. 12a is a fluorescence diagram at 365nm of the graphite oxide phase carbon nitride of the tris-s-triazine structure, the graphite oxide phase carbon nitride of the triazine structure, and the graphite phase carbon nitride of the triazine structure of the present invention;
fig. 12b is a fluorescence emission spectrum of the graphite oxide phase carbon nitride with the triazine structure of the invention under different excitation waves, and fig. 12b shows corresponding test results of 365nm, 380nm, 400nm, 405nm, 410nm, 430nm, 440nm, 450nm, 340nm, 320nm and 300nm from top to bottom.
As shown in FIG. 12a, under 365nm excitation, the graphite phase carbon nitride (tri-s-C) of the tris-s-triazine structure3N4) The emission peak of the catalyst is 443nm, and the modified graphite oxide phase carbon nitride (tri-s-HC) with the tris-s-triazine structure is obtained3N4) The emission peak has a little blue shift (436 nm); and graphite phase carbon nitride (tri-C) of triazine structure3N4) The emission peak of (1) is 497nm, and the modified graphite oxide phase carbon nitride (tri-HC) with triazine structure is obtained3N4) And the emission peak also has a little blue shift (481nm), which shows that after the modification treatment, the conjugated degree of the material structure is reduced due to the appearance of the oxygen-containing functional group, so that the fluorescence blue shift of the modified material is caused. tri-HC obtained after modification3N4The position (481nm) of the emission peak of (A) is lower than that of the modified tri-s-HC3N4(436nm) has larger red shift, and the peak shape becomes more symmetrical, which shows that the trimesic acid is added to successfully convert the triazine structure into the triazine structure, and the conjugation degree is enhanced to cause the fluorescence red shift. In addition, under 365nm excitation, the tri-HC3N4 emits green light, and the tri-s-HC3N4Emitting blue light; as shown in fig. 12b, the fluorescence emission position does not change with the change of the excitation wavelength; under the excitation of 300-340nm, the fluorescence intensity is very weak, and under the excitation of 365-400nm, the fluorescence intensity is weakened along with the increase of the excitation wavelength; the fluorescence is strongest at 365 nm; under the excitation of 400-430nm, the fluorescence intensity is basically consistent and is in medium-intensity fluorescence; the fluorescence intensity becomes weaker under the excitation of 440-450nm, but still higher than that under the excitation of 300-340 nm. The above results show that tri-HC3N4The fluorescence is strongest at 365nm and can emit fluorescence with certain intensity under the irradiation of the visible light region of 400-430nm, which shows that the fluorescence tri-HC3N4The material has practical application value in the fields of biological imaging and biomedicine.
Experimental example 6
This experimental example 6 examined cytotoxicity of graphite oxide phase carbon nitride of triazine structure. L929 Normal cells were cultured at 2X 104Individual cells/well density were plated in 96-well plates with 5% CO at 37 deg.C2Under the conditions of (1) overnight. The graphite oxide phase carbon nitride (tri-HC) prepared in example 1 was added3N4) Set to different concentrations (0, 5, 10, 20, 30, 50, 60, 80, 100,200 and 400. mu.g/mL), added to the well plate, incubated at 37 ℃ for 24h,48h and 72h, finally removed of the medium, washed three times with PBS, added with a concentration of CCK-8 reagent, and measured with a microplate reader under excitation at 450nm, the results of which are shown in FIG. 13.
As shown in FIG. 13, when the concentration is lower than 100. mu.g/mL, the survival rate of the L929 cell is as high as more than 70%, and good cell activity is shown; the survival rate for 72h was 58.12% at a concentration of 200. mu.g/mL, slightly above the limit IC for safety assessment50But with the lapse of culture time, the activity of the cells was significantly reduced; when the concentration is increased to 400. mu.g/mL, the survival rate at 24h is only 50.33%When the survival rate after 72 hours is reduced to 39.26 percent, the traditional Chinese medicine preparation has great toxicity to cells. The above experimental results show that tri-HC3N4The toxicity to cells is concentration-dependent, and the safety of the cells is good in the range of 100 mu g/mL.
This experimental example 6 also examined the effect of the concentration on the intracellular uptake of the graphite oxide phase carbon nitride of the triazine structure.
Cal27 cells were cultured at 2.0X 105Density per well in 6-well plates, 5% CO at 37 ℃2After overnight incubation under the conditions of (1), the medium was discarded, washed three times with PBS, and then used containing the graphite oxide phase carbon nitride (tri-HC) prepared in example 13N4) Different concentrations of tri-HC3N4(0, 10, 20, 40, 60, 80, 100. mu.g/mL) of fresh medium was added to the well plate and after 8h of co-cultivation, cells were resuspended in 500. mu.L of PBSF by trypsinization, centrifugation at 2000rpm for 5min, and then washed three times with PBS. Finally, the mixture is excited by a visible excitation wavelength of 405nm and detected by a flow cytometer, and the detection result is shown in FIG. 14.
As shown in FIG. 14, the amount of the cells was increased with the increase of the concentration under the co-cultivation conditions at the same time, and it is evident from the graph that the amount of the cells was sharply increased from the initial 0.32% to 71.4% at a low concentration (10-40. mu.g/mL), and when the concentration was further increased (60-100. mu.g/mL), the fluorescence of the tri-HC was observed3N4The cell-entering amount is increased smoothly, and reaches 90.4% when the cell-entering amount is 100 mu g/mL, which indicates that the fluorescence tri-HC3N4Can successfully enter cells, and the cell entering amount is concentration-dependent. The results show that the cell-entering amount of the material is over half at 20 mu g/mL, and the cell-entering amount of 40 mu g/mL reaches 71.4 percent, so the fluorescence tri-HC is selected by comprehensively considering the possible toxicity of the material to cells, the caused isohexia reaction and the cell-entering amount of the material3N4The optimal concentration of material was 40. mu.g/mL.
This experimental example 6 also examined the effect of time on intracellular uptake of the graphite oxide phase carbon nitride of the triazine structure. Cal27 cells were cultured at 2.0X 105Density per well in 6-well plates, 5% CO at 37 ℃2Under the conditions ofAfter overnight incubation, the medium was discarded, washed three times with PBS, and then added with a solution containing 40. mu.g/ml tri-HC prepared in example 13N4The cells were resuspended in 500. mu.L of PBSF by incubating the fresh medium for different periods of time (30min, 1h, 2h, 4h and 6h), trypsinized, centrifuged at 2000rpm for 5min, and washed three times with PBS. Finally, the mixture is excited by a visible excitation wavelength of 405nm and detected by a flow cytometer, and the detection result is shown in FIG. 15.
As shown in FIG. 15, the cell-entering amount of the fluorescent material gradually increased with the passage of time, the cell-entering amount reached 53.5% at 4h, and the cell-entering amount at 6h rose more gradually than that at the previous period, indicating that the fluorescence tri-HC3N4The material can enter cells in a short time, and the cell entering amount is increased along with the increase of time.
This experimental example 6 also examined the intracellular fluorescence imaging effect of the graphite oxide phase carbon nitride of the triazine structure. And (3) spreading the sterile cell slide on the bottom of the 24-pore plate, and sterilizing for later use. Cal27 cells were cultured at 5.0X 104Density of/well on 24-well slide plate, after overnight culture, the medium was discarded, washed three times with PBS, and then graphite oxide phase carbon nitride (tri-HC) containing triazine structure of 40. mu.g/mL prepared in example 1 was used3N4) The fresh medium was added to the well plate and incubation continued for 30min, 1h, 2h, and 4h, respectively. Discarding the culture medium, washing with PBS three times, fixing with 4% paraformaldehyde for 15min, washing with PBS three times, staining with DAPI for 10min, washing with PBS three times, sealing with anti-fluorescence quencher, and imaging with laser confocal detection, wherein the test result is shown in FIG. 16, wherein staining of cell nucleus with DAPI shows blue fluorescence, and tri-HC shows blue fluorescence3N4Green fluorescence is emitted.
As shown in FIG. 16, fluorescence tri-HC3N4The material is a carbonaceous material which can successfully enter cells and has a fluorescent color different from that of a cell nucleus stain DAPI, and the cell entering amount is increased along with the increase of time, so that strong green fluorescence is shown. At 0.5h, a weaker green fluorescence appeared, indicating fluorescence of tri-HC over a short period of time3N4The material can pass throughThe cell endocytosis and other ways enter cancer cells, and as the culture time increases, more and more fluorescent tri-HC3N4Material is engulfed and some may enter the nucleus.
Experimental example 7
Experimental example 7 examines tri-HC prepared by the present invention3N4Intracellular fluorescence imaging of DOX. And (3) spreading the sterile cell slide on the bottom of the 24-pore plate, and sterilizing for later use. Cal27 cells were cultured at 5.0X 104Density of/well on 24-well slide glass, after overnight culture, the medium was discarded, washed three times with PBS, and then pure DoX containing the same amount (2. mu.M) of Doxorubicin (DOX), the triazine-structured graphite oxide-phase carbon nitride (tri-HC) prepared in example 1 was used3N4) And doxorubicin-supported triazine structure graphite oxide phase carbon nitride (tri-HC)3N4DOX) was added to the well plates and incubation continued for 4 h. The medium was discarded, washed three times with PBS, then fixed with 4% paraformaldehyde for 15min, washed three times with PBS, stained with DAPI for 10min, washed three times with PBS, mounted with an anti-fluorescence quencher, and imaged with laser confocal detection, with the test results shown in fig. 17.
As shown in FIG. 17a, tri-HC3N4Under the excitation of 405nm, the emission of 481nm is shown, and no emission peak appears under the excitation of 488 nm; DOX in FIG. 17b shows strong emission peaks at 558nm and 587nm at 488nm excitation, while the fluorescence intensity is weaker at 405nm excitation; FIG. 17c shows that upon combination, the emission peak is mainly DOX, and the emission peak is mainly 587nm when excited at 488 nm. According to the above fluorescence property analysis, since the material and the drug DOX have different optimal excitation wavelengths and different emission wavelengths, it is possible to analyze tri-HC3N4The green FITC channel under 405nm excitation is selected for positioning, and the TRITC red channel under 488nm excitation is used for monitoring released DOX, so that material positioning and drug monitoring are carried out simultaneously.
This experimental example 7 also examined the intracellular localization of the graphite oxide-phase carbon nitride of the triazine structure supporting doxorubicin. Passing the sterile cellSpreading the slices on the bottom of 24-well plate, and sterilizing. Cal27 cells were cultured at 5.0X 104Density of/well on 24-well slide glass, after overnight culture, the medium was discarded, washed three times with PBS, and then pure DoX containing the same amount (2. mu.M) of Doxorubicin (DOX), the triazine-structured graphite oxide-phase carbon nitride (tri-HC) prepared in example 1 was used3N4) And doxorubicin-supported triazine structure graphite oxide phase carbon nitride (tri-HC)3N4DOX) was added to the well plates and incubation continued for 4 h. The medium was discarded, washed three times with PBS, then fixed with 4% paraformaldehyde for 15min, washed three times with PBS, stained with DAPI for 10min, washed three times with PBS, mounted with an anti-fluorescence quencher, and imaged with laser confocal detection, with the test results shown in fig. 18.
As shown in FIG. 18, pure tri-HC3N4Blue fluorescence of DAPI and tri-HC at 405nm excitation after Material and cell culture3N4The green fluorescence of the material, no fluorescence was observed under 488nm excitation. As can be seen from FIG. 18, tri-HC3N4The material can enter Cal27 cells, and part of the material can enter cell nucleus, which indicates that the tri-HC3N4The material is a drug transport carrier with a tracing effect, is a potential cell nucleus coloring agent, and provides a harmless carbonaceous material for green dyeing of cell nuclei. The pure DOX image shows that only blue fluorescence of cell nucleus is shown under 405nm excitation, strong red fluorescence and weak green fluorescence are shown under 488nm excitation, which indicates that pure DOX can enter cells, and part of DOX can enter the cells and directly participate in the damage of DNA in the cells, thereby killing the cells. When tri-HC3N4Loading with DOX to tri-HC3N4after-DOX composite, it can be seen from the figure that not only HC can be seen under excitation of 405nm and 488nm, respectively3N4Fluorescence of the material, and released DOX was also observed, indicating tri-HC3N4As the carrier, not only DOX can be smoothly transported into the cell, but also tri-HC can be utilized3N4Fluorescence of the material itselfTracing the three-hydrocarbon mixture to more intuitively carry out tri-HC3N4The distribution of the material in the body is positioned, and a visual means is provided for the research of the metabolism of the material. Furthermore, when tri-HC3N4When the fluorescent powder is used as a carrier, the red fluorescence of DOX is obviously stronger than that of pure DOX, and the tri-HC is shown3N4More DOX released from the DOX composite material enters cells, and the increase of DOX entering nuclei is observed, so that the killing effect on cancer cells is more favorable.
As can be seen from the above examples and experimental examples, the present invention adopts the introduction of trimesic acid to convert the graphite phase carbon nitride with the tris-s-triazine structure into the graphite phase carbon nitride fluorescent material with the triazine structure, and adopts concentrated nitric acid and high temperature and high pressure methods to modify the surface of the graphite phase carbon nitride fluorescent material, so as to prepare the graphite oxide phase carbon nitride fluorescent nanomaterial with the triazine structure. Compared with the existing graphite oxide phase carbon nitride with a triazine structure and the graphite phase carbon nitride with the triazine structure, the graphite oxide phase carbon nitride with the triazine structure shows excellent stability, water dispersibility and biocompatibility under aqueous solution and physiological conditions. In addition, the triazine-structure graphite oxide-phase carbon nitride-loaded doxorubicin hydrochloride can be effectively absorbed by cancer cells and enriched in cell nuclei, and can emit strong green fluorescence under the irradiation of visible light, so that tri-HC can be simultaneously realized3N4Material localization and drug monitoring of DOX. Therefore, the graphite oxide phase carbon nitride provided by the invention has wide application in the fields of drug delivery, biological imaging and tracing, and the anticancer drug provided by the invention also has wide application prospect in preparing drugs for treating malignant tumors or aplastic anemia.
The foregoing is merely exemplary and illustrative of the present invention and it is within the purview of one skilled in the art to modify or supplement the embodiments described or to substitute similar ones without the exercise of inventive faculty, and still fall within the scope of the claims.
Claims (9)
1. A preparation method of graphite oxide phase carbon nitride is characterized by comprising the following steps:
step (1): uniformly mixing guanidine hydrochloride and trimesic acid according to the mass ratio of 3-7: 1, placing the mixture into a reaction container, heating the mixture to 340-360 ℃ at the normal pressure and the normal temperature at the heating rate of 5-7 ℃/min, reacting for 3-5 hours, cooling the mixture to the room temperature, taking out a solid product, grinding the solid product into powder, washing and drying the powder to obtain graphite-phase carbon nitride;
step (2): placing the graphite-phase carbon nitride obtained in the step (1) into a reaction container, adding concentrated nitric acid, refluxing for 70-80 hours at the temperature of 80-95 ℃, then centrifugally washing to be neutral, and drying to obtain porous graphite-phase carbon nitride;
and (3): and (3) placing the porous graphite phase carbon nitride obtained in the step (2) into a reaction kettle, adding distilled water, and reacting at the temperature of 170-200 ℃ for 7-10 hours to obtain the graphite oxide phase carbon nitride.
2. The method for preparing graphite oxide-phase carbon nitride according to claim 1, wherein the mass ratio of guanidine hydrochloride to trimesic acid in the step (1) is 5:1, the temperature increase rate is 5 ℃/min, the reaction temperature is 350 ℃, and the reaction time is 3.5 hours.
3. The method for producing graphite oxide-phase carbon nitride according to claim 1, wherein the reflux temperature in the step (2) is 80 ℃ and the reflux time is 72 hours; the reaction temperature in the step (3) is 180 ℃, and the reaction time is 8 hours.
4. The graphite oxide-phase carbon nitride obtained by the method for producing graphite oxide-phase carbon nitride according to any one of claims 1 to 3.
5. Use of the graphite oxide phase carbon nitride of claim 4 in the preparation of an anti-cancer medicament.
6. An anticancer drug characterized by using the graphite oxide-phase carbon nitride according to claim 4 as a carrier.
7. The anticancer agent as set forth in claim 6, wherein the active ingredient of the anticancer agent is doxorubicin hydrochloride.
8. The process for preparing an anticancer agent according to any one of claims 6 to 7, comprising the steps of:
step (1): respectively dissolving graphite oxide phase carbon nitride and doxorubicin hydrochloride in a phosphate buffer solution with the pH value of 2-11 to obtain a solution A and a solution B;
step (2): and (2) mixing the solution A and the solution B obtained in the step (1) under a dark condition, reacting for 5-24 hours on a shaking table, and then separating, purifying and washing to obtain the anti-cancer drug.
9. The method for preparing the anticancer drug according to claim 8, wherein the mass concentration of the graphite oxide phase carbon nitride in the solution A in the step (1) is 10 to 100 μ g/mL, and the mass concentration of the doxorubicin hydrochloride in the solution B in the step (1) is 5 to 400 mg/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110279315.4A CN113044818A (en) | 2021-03-16 | 2021-03-16 | Graphite oxide phase carbon nitride, anticancer drug, and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110279315.4A CN113044818A (en) | 2021-03-16 | 2021-03-16 | Graphite oxide phase carbon nitride, anticancer drug, and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113044818A true CN113044818A (en) | 2021-06-29 |
Family
ID=76512705
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110279315.4A Pending CN113044818A (en) | 2021-03-16 | 2021-03-16 | Graphite oxide phase carbon nitride, anticancer drug, and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113044818A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050056810A1 (en) * | 2003-09-17 | 2005-03-17 | Jinru Bian | Polishing composition for semiconductor wafers |
| US20100233227A1 (en) * | 2009-03-10 | 2010-09-16 | Boston Scientific Scimed, Inc. | Medical devices having carbon drug releasing layers |
| CN104984349A (en) * | 2015-07-28 | 2015-10-21 | 金陵科技学院 | Epirubicin loaded graphene quantum dot drug carrying system and preparation method thereof |
| CN106238086A (en) * | 2016-07-28 | 2016-12-21 | 华南理工大学 | A kind of phenyl ring modifies class graphite phase carbon nitride photocatalyst and preparation method and application |
| CN109233820A (en) * | 2018-07-16 | 2019-01-18 | 泰山医学院 | A kind of g-C3N4Quantum dot and its preparation method and application |
| CN110327956A (en) * | 2019-06-13 | 2019-10-15 | 福建农林大学 | A kind of preparation method of copper oxide and the compound heterojunction photocatalyst of carbonitride |
-
2021
- 2021-03-16 CN CN202110279315.4A patent/CN113044818A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050056810A1 (en) * | 2003-09-17 | 2005-03-17 | Jinru Bian | Polishing composition for semiconductor wafers |
| US20100233227A1 (en) * | 2009-03-10 | 2010-09-16 | Boston Scientific Scimed, Inc. | Medical devices having carbon drug releasing layers |
| CN104984349A (en) * | 2015-07-28 | 2015-10-21 | 金陵科技学院 | Epirubicin loaded graphene quantum dot drug carrying system and preparation method thereof |
| CN106238086A (en) * | 2016-07-28 | 2016-12-21 | 华南理工大学 | A kind of phenyl ring modifies class graphite phase carbon nitride photocatalyst and preparation method and application |
| CN109233820A (en) * | 2018-07-16 | 2019-01-18 | 泰山医学院 | A kind of g-C3N4Quantum dot and its preparation method and application |
| CN110327956A (en) * | 2019-06-13 | 2019-10-15 | 福建农林大学 | A kind of preparation method of copper oxide and the compound heterojunction photocatalyst of carbonitride |
Non-Patent Citations (4)
| Title |
|---|
| LIYING HAO ET AL: ""Modified triazine-based carbon nitride as a high efficiency fluorescence sensor for the label-free detection of Ag+"", 《JOURNAL OF MATERIALS RESEARCH》 * |
| WEI ZHU ET AL: ""Triazine-based graphitic carbon nitride: controllable synthesis and enhanced cataluminescent sensing for formic acid"", 《ANALYTICAL AND BIOANALYTICAL CHEMISTRY》 * |
| 林祖金 等: ""多孔氢键有机框架（HOFs）:现状与挑战"", 《化学学报》 * |
| 蔡永萍 等主编: "《生物学综合实验教程》", 31 August 2020, 中国农业大学出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dong et al. | Upconversion-mediated ZnFe 2 O 4 nanoplatform for NIR-enhanced chemodynamic and photodynamic therapy | |
| Chen et al. | Stanene‐based nanosheets for β‐elemene delivery and ultrasound‐mediated combination cancer therapy | |
| Jia et al. | Mesoporous cerium oxide-coated upconversion nanoparticles for tumor-responsive chemo-photodynamic therapy and bioimaging | |
| Shu et al. | Fabrication of a hyaluronic acid conjugated metal organic framework for targeted drug delivery and magnetic resonance imaging | |
| Wang et al. | Magnetic iron oxide–fluorescent carbon dots integrated nanoparticles for dual-modal imaging, near-infrared light-responsive drug carrier and photothermal therapy | |
| Yu et al. | Black hollow silicon oxide nanoparticles as highly efficient photothermal agents in the second near-infrared window for in vivo cancer therapy | |
| Wang et al. | Multifunctional PEG encapsulated Fe 3 O 4@ silver hybrid nanoparticles: antibacterial activity, cell imaging and combined photothermo/chemo-therapy | |
| Song et al. | Fabrication of the biomimetic DOX/Au@ Pt nanoparticles hybrid nanostructures for the combinational chemo/photothermal cancer therapy | |
| Han et al. | pH/NIR-responsive nanocarriers based on mesoporous polydopamine encapsulated gold nanorods for drug delivery and thermo-chemotherapy | |
| Chen et al. | Multifunctional Nd 3+-sensitized upconversion nanomaterials for synchronous tumor diagnosis and treatment | |
| Balasamy et al. | Cisplatin delivery, anticancer and antibacterial properties of Fe/SBA-16/ZIF-8 nanocomposite | |
| CN109568578B (en) | Preparation method and application of natural biomass dots and biomass dots-copper nanocomposites | |
| Liu et al. | X-ray and NIR light dual-triggered mesoporous upconversion nanophosphor/Bi heterojunction radiosensitizer for highly efficient tumor ablation | |
| Abdulwahid et al. | Folate decorated dextran-coated magnetic nanoparticles for targeted delivery of ellipticine in cervical cancer cells | |
| Qin et al. | Highly water-dispersible TiO 2 nanoparticles for doxorubicin delivery: effect of loading mode on therapeutic efficacy | |
| CN112641946A (en) | Polydopamine-coated gold nano-composite, preparation method thereof and application thereof in multi-modal tumor diagnosis and treatment | |
| Cheng et al. | 4-in-1 Fe3O4/g-C3N4@ PPy-DOX nanocomposites: magnetic targeting guided trimode combinatorial chemotherapy/PDT/PTT for cancer | |
| Baziulyte-Paulaviciene et al. | Synthesis and functionalization of NaGdF4: Yb, Er@ NaGdF4 core–shell nanoparticles for possible application as multimodal contrast agents | |
| Sun et al. | Stimuli responsive PEGylated bismuth selenide hollow nanocapsules for fluorescence/CT imaging and light-driven multimodal tumor therapy | |
| Liu et al. | Redox/pH-responsive hollow manganese dioxide nanoparticles for thyroid cancer treatment | |
| CN114848813A (en) | A self-supplying H2O2/O2 and GSH-consuming metal-organic framework-coated nanoparticle and its preparation method and application | |
| Yang et al. | Synthesis of boron carbonitride nanosheets using for delivering paclitaxel and their antitumor activity | |
| Hu et al. | An Intelligent and Soluble Microneedle Composed of Bi/BiVO4 Schottky Heterojunction for Tumor Ct Imaging and Starvation/Gas Therapy‐Promoted Synergistic Cancer Treatment | |
| Qin et al. | Fe 3 O 4@ SiO 2 mesoporous spheres as Fe (ii) donors loaded with artemisinin and a photosensitizer to alleviate tumor hypoxia in PDT for enhanced anticancer therapy | |
| CN103768620B (en) | Fe/ mesoporous monox nanometer composite and its preparation method and application |
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 |
Application publication date: 20210629 |
|
| RJ01 | Rejection of invention patent application after publication |