CN112494660A - Preparation method of nano targeted drug and application of nano targeted drug in treatment of gastric cancer - Google Patents
Preparation method of nano targeted drug and application of nano targeted drug in treatment of gastric cancer Download PDFInfo
- Publication number
- CN112494660A CN112494660A CN202011525066.4A CN202011525066A CN112494660A CN 112494660 A CN112494660 A CN 112494660A CN 202011525066 A CN202011525066 A CN 202011525066A CN 112494660 A CN112494660 A CN 112494660A
- Authority
- CN
- China
- Prior art keywords
- nano
- drug
- gnp
- zns
- doxorubicin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000003814 drug Substances 0.000 title claims abstract description 205
- 229940079593 drug Drugs 0.000 title claims abstract description 195
- 238000011282 treatment Methods 0.000 title claims abstract description 50
- 206010017758 gastric cancer Diseases 0.000 title claims abstract description 30
- 208000005718 Stomach Neoplasms Diseases 0.000 title claims abstract description 29
- 201000011549 stomach cancer Diseases 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 230000001225 therapeutic effect Effects 0.000 claims abstract description 127
- 239000002096 quantum dot Substances 0.000 claims abstract description 11
- 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 claims description 329
- 229960004679 doxorubicin Drugs 0.000 claims description 152
- 239000012581 transferrin Substances 0.000 claims description 45
- 102000004338 Transferrin Human genes 0.000 claims description 38
- 108090000901 Transferrin Proteins 0.000 claims description 38
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 30
- 235000018417 cysteine Nutrition 0.000 claims description 30
- 239000000523 sample Substances 0.000 claims description 26
- 230000008685 targeting Effects 0.000 claims description 17
- 229940009456 adriamycin Drugs 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 239000002539 nanocarrier Substances 0.000 claims description 12
- 239000003446 ligand Substances 0.000 claims description 10
- 239000002246 antineoplastic agent Substances 0.000 claims description 9
- 230000015572 biosynthetic process Effects 0.000 claims description 8
- 238000003786 synthesis reaction Methods 0.000 claims description 8
- 229940044683 chemotherapy drug Drugs 0.000 claims description 6
- 229910052984 zinc sulfide Inorganic materials 0.000 abstract description 115
- 239000002105 nanoparticle Substances 0.000 abstract description 29
- 230000000694 effects Effects 0.000 abstract description 27
- 239000002114 nanocomposite Substances 0.000 abstract description 21
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 abstract description 11
- 239000010931 gold Substances 0.000 abstract description 11
- 229910052737 gold Inorganic materials 0.000 abstract description 11
- 231100000331 toxic Toxicity 0.000 abstract description 6
- 230000002588 toxic effect Effects 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 5
- 238000001228 spectrum Methods 0.000 abstract description 5
- DRDVZXDWVBGGMH-UHFFFAOYSA-N zinc;sulfide Chemical class [S-2].[Zn+2] DRDVZXDWVBGGMH-UHFFFAOYSA-N 0.000 abstract description 5
- 238000003745 diagnosis Methods 0.000 abstract description 4
- 238000004393 prognosis Methods 0.000 abstract description 3
- 238000013461 design Methods 0.000 abstract description 2
- 238000011068 loading method Methods 0.000 abstract description 2
- 239000002184 metal Substances 0.000 abstract description 2
- 229910052751 metal Inorganic materials 0.000 abstract description 2
- 239000002086 nanomaterial Substances 0.000 abstract description 2
- 230000005909 tumor killing Effects 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract 1
- 230000001988 toxicity Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 101
- 206010028980 Neoplasm Diseases 0.000 description 62
- 210000001519 tissue Anatomy 0.000 description 59
- 230000027455 binding Effects 0.000 description 40
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 31
- 238000001990 intravenous administration Methods 0.000 description 29
- 201000011510 cancer Diseases 0.000 description 26
- 238000000338 in vitro Methods 0.000 description 26
- 102000005962 receptors Human genes 0.000 description 22
- 108020003175 receptors Proteins 0.000 description 22
- 238000001727 in vivo Methods 0.000 description 19
- 241000699670 Mus sp. Species 0.000 description 17
- 231100000135 cytotoxicity Toxicity 0.000 description 16
- 230000003013 cytotoxicity Effects 0.000 description 16
- 238000000540 analysis of variance Methods 0.000 description 15
- 230000003833 cell viability Effects 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 239000000872 buffer Substances 0.000 description 14
- 238000004128 high performance liquid chromatography Methods 0.000 description 14
- 230000003834 intracellular effect Effects 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 102000007238 Transferrin Receptors Human genes 0.000 description 12
- 108010033576 Transferrin Receptors Proteins 0.000 description 12
- 230000008878 coupling Effects 0.000 description 12
- 238000010168 coupling process Methods 0.000 description 12
- 238000005859 coupling reaction Methods 0.000 description 12
- 239000008367 deionised water Substances 0.000 description 12
- 229910021641 deionized water Inorganic materials 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 11
- 230000004083 survival effect Effects 0.000 description 11
- 229940126585 therapeutic drug Drugs 0.000 description 11
- 230000005284 excitation Effects 0.000 description 10
- 210000002216 heart Anatomy 0.000 description 10
- 210000003734 kidney Anatomy 0.000 description 10
- 206010061968 Gastric neoplasm Diseases 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 8
- 210000004185 liver Anatomy 0.000 description 8
- 210000002784 stomach Anatomy 0.000 description 8
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 230000003247 decreasing effect Effects 0.000 description 7
- 230000008030 elimination Effects 0.000 description 7
- 238000003379 elimination reaction Methods 0.000 description 7
- 238000010253 intravenous injection Methods 0.000 description 7
- 238000005259 measurement Methods 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 6
- 206010009944 Colon cancer Diseases 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 208000029742 colonic neoplasm Diseases 0.000 description 6
- 210000000805 cytoplasm Anatomy 0.000 description 6
- 238000012377 drug delivery Methods 0.000 description 6
- 230000002496 gastric effect Effects 0.000 description 6
- 210000005228 liver tissue Anatomy 0.000 description 6
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 6
- 238000002560 therapeutic procedure Methods 0.000 description 6
- 230000035899 viability Effects 0.000 description 6
- 230000037396 body weight Effects 0.000 description 5
- 231100000433 cytotoxic Toxicity 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 230000004807 localization Effects 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- 230000002378 acidificating effect Effects 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 230000001472 cytotoxic effect Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000003384 imaging method Methods 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 231100000636 lethal dose Toxicity 0.000 description 4
- AOJFQRQNPXYVLM-UHFFFAOYSA-N pyridin-1-ium;chloride Chemical compound [Cl-].C1=CC=[NH+]C=C1 AOJFQRQNPXYVLM-UHFFFAOYSA-N 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 4
- 235000013878 L-cysteine Nutrition 0.000 description 3
- 239000004201 L-cysteine Substances 0.000 description 3
- 238000003917 TEM image Methods 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 238000001802 infusion Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000001509 sodium citrate Substances 0.000 description 3
- 229960003080 taurine Drugs 0.000 description 3
- 206010019695 Hepatic neoplasm Diseases 0.000 description 2
- 208000008839 Kidney Neoplasms Diseases 0.000 description 2
- 231100000111 LD50 Toxicity 0.000 description 2
- 108060001084 Luciferase Proteins 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 150000001718 carbodiimides Chemical class 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 238000000695 excitation spectrum Methods 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 208000024348 heart neoplasm Diseases 0.000 description 2
- 230000002055 immunohistochemical effect Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 238000007726 management method Methods 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 231100000587 neutral red assay Toxicity 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 230000036470 plasma concentration Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 229910052979 sodium sulfide Inorganic materials 0.000 description 2
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000012536 storage buffer Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 238000003325 tomography Methods 0.000 description 2
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 2
- 229940038773 trisodium citrate Drugs 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 230000004584 weight gain Effects 0.000 description 2
- 235000019786 weight gain Nutrition 0.000 description 2
- 239000011592 zinc chloride Substances 0.000 description 2
- 235000005074 zinc chloride Nutrition 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 244000248349 Citrus limon Species 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- UQABYHGXWYXDTK-UUOKFMHZSA-N GppNP Chemical compound C1=2NC(N)=NC(=O)C=2N=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)NP(O)(O)=O)[C@@H](O)[C@H]1O UQABYHGXWYXDTK-UUOKFMHZSA-N 0.000 description 1
- 241000590002 Helicobacter pylori Species 0.000 description 1
- 206010069755 K-ras gene mutation Diseases 0.000 description 1
- 206010024264 Lethargy Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 208000025966 Neurological disease Diseases 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 231100000360 alopecia Toxicity 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 208000002399 aphthous stomatitis Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000030570 cellular localization Effects 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- QBWCMBCROVPCKQ-UHFFFAOYSA-N chlorous acid Chemical compound OCl=O QBWCMBCROVPCKQ-UHFFFAOYSA-N 0.000 description 1
- 229940077239 chlorous acid Drugs 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 239000000599 controlled substance Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000003235 crystal violet staining Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 208000010643 digestive system disease Diseases 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 238000009513 drug distribution Methods 0.000 description 1
- 239000013583 drug formulation Substances 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 229940014144 folate Drugs 0.000 description 1
- 102000006815 folate receptor Human genes 0.000 description 1
- 108020005243 folate receptor Proteins 0.000 description 1
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 208000010749 gastric carcinoma Diseases 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000002343 gold Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229940037467 helicobacter pylori Drugs 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000171 higher toxicity Toxicity 0.000 description 1
- 238000010562 histological examination Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000002595 magnetic resonance imaging Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002082 metal nanoparticle Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000002600 positron emission tomography Methods 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000015598 salt intake Nutrition 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- SDKPSXWGRWWLKR-UHFFFAOYSA-M sodium;9,10-dioxoanthracene-1-sulfonate Chemical compound [Na+].O=C1C2=CC=CC=C2C(=O)C2=C1C=CC=C2S(=O)(=O)[O-] SDKPSXWGRWWLKR-UHFFFAOYSA-M 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 201000000498 stomach carcinoma Diseases 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000010301 surface-oxidation reaction Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 231100000164 trypan blue assay Toxicity 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—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
- A61K47/54—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 organic compound
- A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
-
- 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
- A61K47/62—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 a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/644—Transferrin, e.g. a lactoferrin or ovotransferrin
-
- 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
- A61K47/62—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 a protein, peptide or polyamino acid
- A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6923—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
-
- 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
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0056—Peptides, proteins, polyamino acids
-
- 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
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0065—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the luminescent/fluorescent agent having itself a special physical form, e.g. gold nanoparticle
- A61K49/0067—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the luminescent/fluorescent agent having itself a special physical form, e.g. gold nanoparticle quantum dots, fluorescent nanocrystals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nanotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Materials Engineering (AREA)
- General Physics & Mathematics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Biophysics (AREA)
- Inorganic Chemistry (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medical Informatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Composite Materials (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Manufacturing & Machinery (AREA)
- Optics & Photonics (AREA)
- Ceramic Engineering (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to the technical field of biological medicines, in particular to a preparation method of a nano-targeting drug and application of the nano-targeting drug in treatment of gastric cancer. Gold Nanoparticles (GNPs) are the most compatible nanostructures, which have advantages over other metal nanocomposites, and zinc sulfide quantum dots (ZnS QDs) are very important quantum dot nanoparticles, which are low in toxicity, friendly to humans, and narrow in spectrum band. The two prepared therapeutic nano-drugs have good stability, high drug loading rate and excellent slow release property, can enhance the targeted concentration of the drugs, enhance the treatment effect of tumor killing and reduce the non-specific toxic and side effects, and the design modes of the two drugs can be used for diagnosis, treatment and prognosis of gastric cancer, thereby having good application prospect.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a preparation method of a nano-targeting medicine and application of the nano-targeting medicine in treating gastric cancer.
Background
Among all terrible diseases, cancer is the leading cause of death worldwide and their incidence is increasing worldwide. Among the different types of cancer, stomach cancer is one of the most common cancers with the highest mortality rate, and is probably caused by diet, organic solvents, pesticides and pesticides, and is also directly associated with many factors, such as high salt intake in diet or microbial infection of helicobacter pylori (h. Currently, various imaging techniques are being used for cancer diagnosis, such as single photon emission tomography, Magnetic Resonance Imaging (MRI), X-ray based computer-assisted tomography (CT), and Positron Emission Tomography (PET), all of which are non-invasive imaging methods for cancer detection. Pharmaceutical methods for cancer treatment include chemotherapy, radiotherapy and surgery, and unfortunately, all of these treatments bring about many side effects, and these treatments not only cause diseases such as alopecia, nausea, fatigue, digestive system diseases, canker sores and neurological diseases, but also destroy normal tissues and the cancer cells thereof are not completely eradicated.
In contrast to these therapies, nanotechnology promises the selective delivery of cancer cell-targeting drugs with the help of targeted therapies to eliminate all these side effects. The main objective of targeted drug delivery is to avoid the toxic effects of the drug on normal cells, since another advantage of targeted drug delivery compared to target cells is the limited, centrally controlled drug concentration released. Controlled release of the drug not only expands the efficacy of the drug, but also avoids life loss caused by all adverse reactions related to the drug. Many successful approaches to targeted drug delivery have been published involving different strategies against cells and organs, and most reports are targeted delivery approaches against cancer, targeting their cells using specific membrane-bound antigens of the cancer.
The nano biotechnology brings new scientific prospects for targeted drugs and drug therapy. The involvement of nanocarriers greatly changes the fate of targeted drug delivery, not only affecting the therapeutic field but also the diagnostic and prophylactic fields. Decades of research have demonstrated that biodegradable nanoparticles are the best and friendly bioengineering for drug delivery or for use in target tissues. Researchers have focused on those nanoparticles that are biodegradable and excreted from the body without any harm to the body, and by combining nanotechnology with drugs, called nanomedicines, targeted delivery to cancer cells can minimize cytotoxicity of normal tissues for the action of anticancer agents by aiming to provide maximum dose to the affected site and enhance their biodistribution to target sites.
With the development of nano-drug formulations and the use of metal nanoparticles, substances such as iron, silver, gold or quantum dots have advantages in delivery and imaging. The use of bioluminescent nanoparticles as drug-carrying and targeting nanocarriers to receptor site ligands can provide diagnostic and therapeutic effects, and these nanomedicines with diagnostic and therapeutic functions are called therapeutic nanomedicines.
The main step in the selection of nanocarriers is the selection of cancer cells targeted by appropriate ligands. Expression of many receptors, such as folate and transferrin receptors, is increased many-fold on cancer cells compared to normal humans. Increased expression of transferrin receptor is observed on cancer cells to increase iron uptake in cancer cells to facilitate biological activities for the following reasons, while expression of transferrin receptor is relatively limited on normal cells, and thus transferrin receptor can be used as a targeting site for nano-drugs for drug delivery targeting cancer.
Doxorubicin, although it has a high antitumor activity, cannot distinguish between normal cells and cancer cells. Adriamycin (an anticancer drug) has a low osmotic molecular weight, is easily permeated by all cells, and is as harmful to the health of normal cells. Overcoming the obstacles Adriamycin has a better effect as an anti-cancer agent, which can be combined with Adriamycin.
The therapeutic effect of doxorubicin at specific surface receptors can be enhanced by targeting cancer cells with ligands. The expression of transferrin receptor has attracted the interest of researchers' delivery on cells that target the rapid growth of drugs. Therapeutic agents and targeting ligands are determined, and the main problems are the selection of nano-carriers, and the selection of Gold Nanoparticles (GNPs) and zinc sulfide quantum dots (ZnS QDs) as nano-carriers to be applied to cancer diseases, which are not reported in the open literature.
Disclosure of Invention
The invention aims to provide a preparation method of a nano-targeted drug and application of the nano-targeted drug in treatment of gastric cancer.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a preparation method of a nano-targeted drug and application of the nano-targeted drug in the treatment of gastric cancer, wherein the nano-targeted drug comprises a GNP therapeutic nano-drug and a ZnS QD therapeutic nano-drug.
The linker of the GNP therapeutic nano-drug is cysteine, targeting ligand transferrin, chemotherapeutic drug adriamycin, and the nano-carrier is GNP.
The linker of the ZnS QD therapeutic nano-drug is cysteine, the targeting ligand transferrin, the chemotherapeutic drug doxorubicin, and the nano-carrier is ZnS QD.
The invention provides a preparation method of a nano-targeted drug, which comprises the preparation of a GNP therapeutic nano-drug and the preparation of a ZnS QD therapeutic nano-drug.
The preparation of GNP therapeutic nano-medicine includes the synthesis of GNP, the combination of cysteine and GNP, the combination of adriamycin and GNP-Cys, and the combination of transferrin and GNP-Cys-Dox.
The preparation of the ZnS QD therapeutic nano-drug comprises the synthesis of ZnS QD quantum dots coated by cysteine, the combination of adriamycin and ZnS QD-Cys, and the combination of transferrin and ZnS QD-Cys-Dox.
Conjugation of cysteine-coated GNPs to transferrin can serve as targeted diagnostic probes.
The invention has the beneficial effects that:
in the present invention, two therapeutic nano-drugs are prepared, and Gold Nanoparticles (GNPs) as nano-carriers are the most compatible nanostructures, which can be used for diagnosis, prognosis, imaging and therapy, with greater advantages than other metal nanocomposites. Zinc sulfide quantum dots (ZnS QDs) are very important quantum dot nanoparticles, and have the advantages of low toxicity, narrow spectrum band, adjustable emission profile size and high emission quantum yield. The two prepared therapeutic nano-drugs have good stability, high drug loading rate and excellent slow release property, can enhance the targeted concentration of the drugs, enhance the treatment effect of tumor killing and reduce the non-specific toxic and side effects, and the design modes of the two drugs can be used for diagnosis, treatment and prognosis of gastric cancer, thereby having good application prospect.
Drawings
FIG. 1 is a UV-VIS spectrum, (a) a UV-VIS spectrum of GNP-Cys, and (b) a UV-VIS spectrum of transferrin-conjugated GNP-Cys.
FIG. 2 is an excitation and emission spectrum of a target diagnostic probe.
Figure 3 is a TEM image of GNP-Cys and transferrin-bound GNP-Cys.
FIG. 4 is a full tumor histological examination and histological study of tumor tissue after binding of a targeted diagnostic probe (in vivo). (a) Bright field studies of tumor tissue show the morphology of the tissue; (b) examination of the whole tissue under an ultraviolet-activated filter showed in vivo binding of the target probe to transferrin receptor on tumor cells; (c) whole tissue examination under blue excitation filters; (d) and (e) histological studies of tumor cryostat sections were identical to those observed in whole tissues.
FIG. 5 is a histological study of in vitro total stomach tissue examination and in vivo tissue-targeted diagnostic probe binding. (a) Bright field studies of stomach tissue have shown tissue morphology with regions of mass; (b) in the whole tissue examination under the ultraviolet ray excitation filter, the target probe is combined with transferrin receptor in a caking area in vivo; (c) the blue excitation filter under examination of the tissue shows that the tissue fluoresces more in this region than in the ultraviolet; (d) and (e) histological studies of gastric carcinoma tissue cryostat sections showed the same results as observed throughout the tissue examination.
FIG. 6 is an in vitro binding of histological studies of tumor and gastric cancer tissue sections to a targeted diagnostic probe. (a) And (b) binding of a diagnostic probe to the tumor tissue section; (c) and (d) binding of the targeted diagnostic probe to the stomach tissue section.
Fig. 7 is a table of cytotoxicity of different GNP nanocomplexes.
Fig. 8 is a graph of the cytotoxicity effect of different GNP nanocomplexes.
Figure 9 is a table of cytotoxicity of different concentrations of GNP therapeutic nanomedicines.
Figure 10 is a graph of the cytotoxicity of GNP therapeutic nanomedicines at different concentrations. (a) The cumulative effect of the nano-drugs with different concentrations in 6 and 24 hours; (b) cytotoxic effects at different concentrations at 6 and 24 hours at the two time points.
Fig. 11 is a table of the cytotoxicity of GNP tumor treatment nano-drugs at different time points.
Figure 12 is a study of GNP nanocomposite growth, cell morphology and fluorescence at the 6 hour time point.
Figure 13 is a study of GNP nanocomposite growth, cell morphology and fluorescence at the 12 hour time point.
Figure 14 is a study of GNP nanocomposite growth, cell morphology and fluorescence at the 24 hour time point.
Fig. 15 is a study of GNP nanocomposite growth, cell morphology and fluorescence at the 48 hour time point.
Fig. 16 is a graph of the viability and percentage of live/dead cells of GNP nanocomplexes at different time points. (a) The GNP tumor treatment nano-drug has cytotoxic effect in 6, 12, 24, 48 and 72 hours; (b) the cytotoxic effect of GNP therapeutic nanomedicines causes a change in the proportion of live and dead cells at 6, 12, 24, 48, 72 hours.
Fig. 17 is an intracellular tracking of GNP nanocomplexes as GNP and doxorubicin at different time points.
Fig. 18 is a fluorescence intensity measurement of GNP therapeutic nanomedicines at various time points.
Fig. 19 is a graph of the concentration of GNP synthesized nanocomplex measured at various time points based on fluorescence intensity.
Figure 20 is in vitro receptor binding of GNP therapeutic nanomedicines on gastric cancer tissues.
Fig. 21 is a cell trace of GNP treatment nanomedicines at different time points.
Fig. 22 shows the fluorescence intensity of GNP therapeutic nanomedicines at different time points.
Figure 23 is a comparison of fluorescence intensity (full window) at different time points after oral administration of GNP therapeutic nanomedicines.
Fig. 24 shows the anti-gastric cancer effect of GNP tumor treatment nano-drug during in vivo treatment.
Figure 25 is the survival rate of mice at different times during 40 days of GNP tumor treatment with the nano-drug.
Figure 26 is a graph of the weight change of live mice over different times during 40 days of GNP therapeutic nano-drug treatment.
Fig. 27 is a table of cytotoxicity of different ZnS nanocomplexes.
Fig. 28 is a table of cytotoxicity of various concentrations of ZnS QD for treatment of nano-drugs.
Fig. 29 is a graph of the cytotoxicity of ZnS QD nanocomposites at different concentrations.
Fig. 30 is a table of cytotoxicity of ZnS QD therapeutic nanomedicines at different time points.
Fig. 31 shows ZnS QD nanocomplex growth at the 6 hour time point, cell morphology and fluorescence studies.
Fig. 32 shows ZnS QD nanocomplex growth at a 12 hour time point, cell morphology and fluorescence studies.
Fig. 33 shows ZnS QD nanocomplexes grown at 24 hour time point, cell morphology and fluorescence studies.
Fig. 34 shows ZnS QD nanocomplexes grown at 48 hour time point, cell morphology and fluorescence studies.
Fig. 35 is a graph of the survival and cytotoxic effects of viable/dead cells of ZnS QD nanocomplexes at different time points.
Fig. 36 is an intracellular tracking of ZnS QD choroidal nanocomplexes as ZnS QDs and doxorubicin at different time points.
Fig. 37 is a measurement of fluorescence intensity of ZnS QD therapeutic nano-drugs at different time points.
FIG. 38 shows fluorescence intensity of ZnS QD nano-compounds at different time points.
Figure 39 is the in vitro receptor binding of ZnS QD therapeutic nanomedicines in gastric cancer tissues.
Fig. 40 is a cell trace of ZnS QD therapeutic nano-drugs at different time points.
FIG. 41 is a comparison of fluorescence intensity at different time points after oral administration of ZnSQD choroidal Nanogen.
Fig. 42 shows the fluorescence intensity of ZnS QD therapeutic nanomedicines at different time points.
Fig. 43 shows the anti-gastric cancer efficacy of ZnS QD therapeutic nano-drugs (in vivo).
Fig. 44 is the survival of mice at various times during 40 days of treatment with ZnS QD therapeutic nanomedicine.
Figure 45 is the change in body weight of live mice for ZnS QD nanotherapeutic drugs at different times during 40 days of treatment.
Fig. 46 shows the anti-gastric cancer efficacy of ZnS QD nano therapeutic drugs (in vivo).
Fig. 47 is a graph of doxorubicin release behavior from GNP therapeutic nanomedicines at pH 5.
Fig. 48 is a graph of doxorubicin release behavior from GNP therapeutic nano-drugs at pH 7.5.
Figure 49 is a graph of release of doxorubicin by GNP therapeutic nanomedicines at pH5 at various time points.
Figure 50 is a graph of release of doxorubicin by GNP therapeutic nanomedicines at pH7.5 at various time points.
FIG. 51 is a chromatogram of doxorubicin.
Fig. 52 is a graph of doxorubicin concentration in the GNP therapeutic nanomedicine targeting response versus non-targeting response.
FIG. 53 is an analysis of the doxorubicin concentration in dead cells and in culture medium.
Figure 54 is the in vitro concentration of doxorubicin in the culture medium and dead cells (GNP therapeutic nano-drug) after different time points.
Figure 55 is the concentration of doxorubicin in the medium (GNP therapeutic nanomedicine) at different time points of incubation.
Fig. 56 is plasma drug activity profile data of GNP therapeutic nano-drugs analyzed by HPLC after intravenous (in vivo) administration.
Figure 57 is a pharmacokinetic parameter for GNP therapeutic nano-drugs.
Fig. 58 is plasma drug activity profile data of GNP therapeutic nano-drugs analyzed by HPLC after oral (in vivo) administration.
Fig. 59 is a plasma drug activity profile of doxorubicin (GNP therapeutic nano-drug) after oral administration.
Fig. 60 is a pharmacokinetic parameter after oral administration of GNP therapeutic nano-drug.
Fig. 61 is a graph of doxorubicin concentration in the liver following intravenous and oral administration of GNP therapeutic nanomedicines.
Fig. 62 is a comparison of the biodistribution of doxorubicin in the liver in the GNP therapeutic nanomedicines at various time points after intravenous and oral administration.
Figure 63 is the cardiac doxorubicin concentration after intravenous and oral administration of GNP therapeutic nano-drug.
Fig. 64 is a comparison of the biodistribution of doxorubicin in the GNP therapeutic nanomedicine in the heart at various time points after intravenous and oral administration.
Figure 65 is the adriamycin concentration in the kidney after intravenous and oral administration of GNP therapeutic nanomedicines.
Fig. 66 is a comparison of the biodistribution of doxorubicin in the GNP therapeutic nanomedicine in the kidney at various time points after intravenous and oral administration.
Fig. 67 is a comparison of the biodistribution of the GNP therapeutic nano-drug doxorubicin in gastric tumors at different time points after intravenous and oral administration.
Figure 68 is the concentration of doxorubicin in gastric tumors following intravenous and oral administration of GNP therapeutic nanomedicines.
Figure 69 is the release behavior of doxorubicin from ZnS QD therapeutic nano-drug at pH 5.
Fig. 70 is a graph of doxorubicin release behavior from ZnS QD therapeutic nano-drug at pH 7.5.
Fig. 71 shows release of doxorubicin from ZnS QD therapeutic nanomedicine at pH5 at different time points.
Fig. 72 shows release of doxorubicin from ZnS QD therapeutic nanomedicine at pH7.5 at different time points.
Figure 73 is the doxorubicin concentration in ZnS QD therapeutic nanomedicine targeting response versus non-targeting response.
FIG. 74 is an analysis of doxorubicin concentration in dead cells and medium (non-targeted versus targeted delivery).
Figure 75 is the in vitro concentration of doxorubicin in the culture medium and in dead cells (ZnS QD therapeutic nano-drug) after different time points.
Figure 76 is the concentration of doxorubicin in the medium (ZnS QD therapeutic nano-drug) at different time points of incubation.
Fig. 77 is in vivo plasma drug activity profile data of nano-drugs measured by HPLC after ZnS QD intravenous administration.
Fig. 78 is a plasma drug activity profile of doxorubicin (ZnS QD therapeutic nano-drug) after intravenous administration.
Fig. 79 shows pharmacokinetic parameters of ZnS QD therapeutic nanomedicines.
Fig. 80 is data of plasma drug activity spectra analyzed by HPLC after oral administration of ZnS QD-based nano-drugs.
Figure 81 is a plasma drug activity profile of doxorubicin (ZnS QD therapeutic nano-drug) after oral administration.
Fig. 82 is the pharmacokinetic parameters of ZnS QD therapeutic nanomedicines after oral administration.
Fig. 83 is the concentration of doxorubicin in the liver following intravenous and oral administration of ZnS QD therapeutic nano-drugs.
Fig. 84 is a comparison of the biodistribution of doxorubicin in the liver in ZnS QD therapeutic nanomedicine at different time points after intravenous and oral administration.
Fig. 85 is a graph of doxorubicin concentration in the heart after intravenous and oral administration of ZnS QD therapeutic nanomedicine.
Fig. 86 is a comparison of the biodistribution of doxorubicin in the heart in ZnS QD therapeutic nanomedicine at different time points after intravenous and oral administration.
Figure 87 is the concentration of doxorubicin in the kidney following intravenous and oral administration of ZnS QD therapeutic nanomedicine.
Fig. 88 is a comparison of the biodistribution of doxorubicin in the kidney for ZnS QD therapeutic nanomedicine at different time points after intravenous and oral administration.
FIG. 89 comparison of the biodistribution of ZnS QD therapeutic NanoTamycin in gastric tumors at different time points after intravenous and oral administration
Fig. 90 is a graph of doxorubicin concentration in gastric tumors after intravenous injection and oral administration of ZnS QD therapeutic nano-drugs.
Detailed Description
The invention is further illustrated by the following examples in conjunction with the accompanying drawings, which are not intended to limit the scope of the invention.
EXAMPLE 1 preparation of Nanotherapeutic drugs
Two kinds of nano medicine are prepared separately. Both nano-therapeutic drugs have the same linker: cysteine, the targeting ligand is transferrin, and the chemotherapeutic drug is adriamycin, but the nano-carriers are different. Gold Nanoparticles (GNPs) were used in the first nanotherapeutic drug due to their unique physical properties, zinc sulfide quantum dots (ZnS) were selected as the second nanotherapeutic drug nanocarriers.
The preparation of the nano therapeutic medicine is as follows:
preparation of GNP nano-therapeutic medicine, including synthesis of GNP, combination of cysteine and GNP, combination of adriamycin and GNP-Cys, and combination of transferrin and GNP-Cys-Dox.
GNPs were synthesized using an optimized citric acid reduction method, gold salts and sodium citrate at different concentrations were used to check the ideal concentrations, GNPs showed different optical properties, and HAuCl was prepared using a 1% gold salt stock solution (chlorous acid) in order to optimize the final reaction4And diluted to water in a 15mM working solution beaker in 400ml of deionized water, the solution was continuously stirred and kept boiling on a hot plate preset at 100 ℃. Preparation of 1% lemon in deionized WaterThe trisodium citrate solution is heated to 50 ℃ and 12.5ml of preheated 1% trisodium citrate are rapidly added to the boiling solution with vigorous stirring, stirring is continued and observed until the color of the reaction mixture becomes dark. The solution turned bright red with a reaction time of about 20 minutes, and the bright red color showed a size of less than 15nm, completing the synthesis of GNP.
An L-cysteine solution (0.2mM) was prepared in deionized water, and 1ml was added to the prepared 400ml GNP solution. The solution was stirred at 25 ℃ for 2 hours and then shielded for 12 hours without interference. Cysteine-attached gold nanoparticles were centrifuged and run at 14,000rpm for 40 minutes. The precipitate was washed with deionized water.
Doxorubicin and cysteine attached gold nanoparticles were purified by the (1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide (EDC) method, 50mg cysteine coated 1ml gnp deionized water, adjusted with 1M NaOH at pH 81 ml EDC (20mg/ml added deionized water) to activate the carboxyl group, and adjusted to pH6.4 with 1M HCl, the reaction mixture was incubated in the dark for 30 minutes with continuous shaking at 100rpm, after 30 minutes of incubation, 1ml carbodiimide (10mg/ml deionized water) was added to the drug containing 50mg doxorubicin and incubated for a further 2 hours in a dark environment at 37 ℃, continuously shaken at 100rpm, centrifuged to separate the conjugate particles at 14000rpm for 20 minutes, and washed twice with deionized water and stored at 4 ℃.
Transferrin is conjugated to doxorubicin conjugated cysteine coated GNP nanocomplexes by the glutaraldehyde method. Briefly, the amine group of cysteine on gold nanoparticles (GNP-Cys) was crosslinked with the amine group of transferrin by glutaraldehyde crosslinking. 100mg of GNP-Cys-Dox was suspended in 100ml of coupling buffer (0.01M pyridine hydrochloride, pH6, 0.1M NaCl) and sonicated for 10 min. After 10 minutes, the suspension was centrifuged at 14000rpm for 40 minutes and then the supernatant was aspirated. The above procedure was repeated twice with coupling buffer and was performed after the second wash. The nanocomposite was suspended in ligation buffer (5% glutaraldehyde in coupling buffer), the suspension was shaken on an orbital shaker at 25 ℃ for 3 hours at 100rpm, and then centrifuged again at 14000rpm for 20 minutes. The supernatant was aspirated and washed with the nanocomplexes with coupling buffer as described previously to remove excess glutaraldehyde. 10mg transferrin was dissolved in 100ml coupling buffer and left to stand in 1ml pre-coupling buffer. The remaining transferrin solution was mixed with 100mg of washing solution to mix the nanocomposite. The mixture was shaken at 25 ℃ for 24 hours and then centrifuged at 8000rpm at 4 ℃ to obtain transferrin-coupled GNP-Cys-Dox nanocomplexes (GNP therapeutic nanomedicines (GNP-Cys-Dox-transferrin) — the conjugated nanocomplexes were suspended in storage buffer (0.01M Tris-Cl, ph7.4, 0.1% sodium azide, 0.15M NaCl) and stored at-20 ℃.
The preparation of ZnS QD nano therapeutic drug comprises the synthesis of ZnS QD quantum dots coated by cysteine, the combination of adriamycin and ZnS QD-Cys, and the combination of transferrin and ZnS QD-Cys-Dox.
Cysteine-coated zinc sulfide quantum dots (znqd) were synthesized in the presence of L-cysteine and taurine by reacting zinc chloride with sodium sulfide in water, briefly, 0.01M zinc chloride solution at pH8, 0.01M L-cysteine solution at pH8 and 0.1M taurine were refluxed for half an hour in a total reaction volume of 200 ml. Taurine is added as an antioxidant to prevent surface oxidation of quantum dots synthesized in air and to improve the size and fluorescent quality of quantum dots prepared in air. After refluxing for half an hour, 0.01M sodium sulfide in 10ml deionized water was added dropwise to the reaction mixture and refluxed for a further 12 hours. The synthesized cysteine-coated ZnS QD nanoparticles were run centrifugally at 14000rpm for 20 minutes at 25 ℃. The precipitate was washed with deionized water.
Doxorubicin and cysteine-coated ZnS QD were reacted by the (1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide (EDC) method, 100mg of cysteine-coated ZnS was substituted for GNP, adjusted with 1M NaOH pH8, 1ml of EDC (20mg/ml added deionized water) to activate carboxyl groups, the pH was adjusted to pH6.4 with 1M HCl the reaction mixture was incubated for 30 minutes in the dark, and continuously shaken at a rotation speed of 100rpm, after incubating for 30 minutes, 1ml of carbodiimide (10mg/ml deionized water) was added to the drug containing 50mg of doxorubicin, and further incubated for 2 hours at 37 ℃ in the dark, the nanocomposite conjugated to doxorubicin was isolated by centrifugation at 14000rpm for 20 minutes and washed twice with deionized water, and the washed nanocomposite was stored at 4 ℃.
The glutaraldehyde method couples transferrin to doxorubicin-coupled cysteine-coated ZnS QDs. 100mg ZnSQD-Cys-Dox was suspended in 100ml of coupling buffer (0.01M pyridine hydrochloride, pH6, 0.1M NaCl) and sonicated for 10 min. After 10 minutes, the suspension was centrifuged at 14000rpm for 40 minutes and then the supernatant was aspirated. The above procedure was repeated twice with coupling buffer and was performed after the second wash. The nanocomposite was suspended in ligation buffer (5% glutaraldehyde in coupling buffer), the suspension was shaken on an orbital shaker at 25 ℃ for 3 hours at 100rpm, and then centrifuged again at 14000rpm for 20 minutes. The supernatant was aspirated and washed with the nanocomplexes with coupling buffer as described previously to remove excess glutaraldehyde. Transferrin (10mg) was dissolved in 100ml of coupling buffer and left to stand in a 1ml pre-coupling buffer. The remaining transferrin solution was mixed with 100mg of washing solution to mix the nanocomposite. The mixture was shaken at 25 ℃ for 24 hours and then centrifuged at 8000rpm at 4 ℃ to obtain transferrin-coupled ZnS QD-Cys-Dox nanocomplex (ZnS QD-Cys-Dox-transferrin) the conjugated nanocomplex was added to storage buffer (0.01M Tris-Cl, ph7.4, 0.1% sodium azide, 0.15M NaCl) and stored at-20 ℃.
Example 2 preparation and characterization of diagnostic probes
Synthetic cysteine-coated GNPs are conjugated to transferrin for diagnostic detection. It is used to examine the efficiency of transferrin as a targeting ligand on cancer cells and hepatoma cells. Transferrin was also examined for expression in tissues that induce gastric cancer.
As shown in fig. 1(b), UV-Vis spectra of the target diagnostic probes were compared with UV-Vis spectra of cysteine-coated GNPs, as shown in fig. 1 (a). The spectrum of the cysteine-coated GNPs showed a peak at 520nm, indicating a particle size of less than 20nm, narrow size distribution. After transferrin binding, the new spectrum shows a shift of the inorganic peak at 525nm and the organic peak at 340nm, as shown in FIG. 1 (b). The peak becomes broader, indicating that conjugation of the protein on the nanoparticle results in an increase in the size of the nanocomposite.
By fixing a drop of diagnostic probe on a glass slide and observing under a fluorescence microscope. The nanocomposites appear blue under uv excitation, and in blue excitation filters they appear green. As shown in FIG. 2, excitation and emission spectra of the target diagnostic probe were obtained, which showed a maximum excitation at 361.0nm and a maximum emission at 362.4 nm.
Hydrodynamic particle size distribution and target mean diameter diagnostic probes were measured by DLS at 25 ℃. The particle size distribution is 100nm to 110nm in the solution, and the average diameter of the monodisperse particles is 100 nm.
TEM micrographs of cysteine coated gold nanoparticles and targeted diagnostic probes showed an increase in size upon transferrin binding. Cysteine-coated TEM micrographs GNPs exhibited spherical nanoparticles with an average diameter of 10nm, as shown in fig. 3 (a). Transferrin binding does not affect shape but results in a 25nm increase in size with an average diameter of 35nm, as shown in figure 3 (b).
The receptor binding ability of purified transferrin conjugated cysteine coated GNPs further demonstrates the specific binding of the probe and its potential application as a targeted diagnostic probe in vivo and in vitro. Ex vivo and ex vivo histological studies of gastric cancer and tumor tissues showed that the probes bound to respective receptors on cancer cells and generated fluorescence images under respective filters. Fig. 4(a) shows tumor tissue morphology under field microscopy. As shown in fig. 4(b) and 4(c), tumor tissue showed strong fluorescence under both filters after in vivo administration, and binding of the diagnostic probe to tumor cells was confirmed. As shown in fig. 4(d) and 4(e), histological studies of tumor cryostat sections revealed the same results as observed in whole tissues, which further confirmed the results of the tissue examination. After one hour of oral administration, stomach tissue was examined under a gastroscope. Fluorescence microscopy confirmed binding of the probe to the infected cancer as shown in fig. 5(b) and 5(c), and histological studies this section further confirmed the results as shown in fig. 5(d) and 5 (e). In vitro screening also showed that even after color fixation, probes were able to image in vivo with receptor binding even after fixation.
The transferrin receptor on tissue targeted diagnostic probes was further confirmed, binding in vitro and fluorescence studies showed successful binding to fixed tissue even with probes on the receptor. As shown in fig. 6(a) and 6(b), tumor tissue showed slightly different fluorescence patterns ex vivo in vitro compared to in vitro. In vitro, the binding of the targeted diagnostic probe to stomach tissue showed mainly gold fluorescence and blue and green color as shown in FIGS. 6(c) and 6 (d). This further confirms the potential use of transferrin conjugated cysteine coated GNPs as in vitro targeted diagnostic probes for cancer.
Example 3 therapeutic Effect of GNP and ZnS QDs
GNP tumor treatment nano-drugs (in vitro)
Cellular uptake, therapeutic efficacy and cellular targeting of therapeutic Nanoparticulates to colon cancer cells (HCT 116-Luc 2)And confirmed by different analyses. Techniques for studying cell viability intracellular nano-drug tracking include inverted microscopy, fluorescence microscopy, trypan blue assay, neutral red assay (metabolic activity), crystal violet staining assay (cell adhesion studies) and fluorescence intensity measurements. All results were statistically compared to give clear results.
Human colon cancer cell lines: HCT-116 (male patient-derived cell line) was used to examine the effect of therapeutic nano-drugs in vitro. The cell line specification involved p53 wild-type gene and K-Ras gene mutation mismatch defect repair system. The morphology of the cells is spindle-shaped in appearance. The remaining vial was removed from the-80 ℃ freezer and thawed prior to use. Memory (High glucose) Medium with streptomycin and pencillin 100mg/ml solutionAnd 10% heat-inactivated fetal bovine serumHCT-116 cells in the presence of 5% CO2Growth was carried out in a humidified chamber with air and 87% humidity at 37 ℃. Throughout the drug analysis, the culture should be routinely performed. The medium was changed every 2 days and subcultured after 5 days.
Three GNP nanocomposites, lower concentrations of 5 μ Ι (5 μ g/μ Ι), GNP-Cys-Dox and GNP-Cys-Dox-transferrin, were tested against colon cancer cell lines and compared by statistical analysis (ANOVA) to observe the therapeutic effect of the nanocomposites, as shown in figure 7. GNP values have a toxic effect on cancer cells, but only 8% of cells have 92% viability. GNP-Cys-Dox demonstrated higher toxicity due to doxorubicin binding and cell viability, 83%, as shown in fig. 8, which shows cell viability within 6 hours for the first complex (GNP-Cys), the second complex (GNP-Cys-Dox) and the third complex (final therapeutic nanomedicine (GNP-Cys-Dox-Trans)). Mean (n-3) ± SD, p <0.0001 for all groups analyzed, and p <0.001 between the second and third synthesis. After transferrin binding, minor differences in cell viability were observed. GNP tumor treatment nano-drugs showed 80% cell viability and observation of cell morphology results showed that even adherent cells were not in good condition due to the longer residence time of the nano-drug. The data are within the confidence interval, showing the significance of the comparison.
The cytotoxicity of GNP at different concentrations, therapeutic nano-drug of GNP at different concentrations, is equivalent to examining the effect of doxorubicin at different concentrations on colon cancer cells at 6 hours and 24 hours. The present invention is used to examine the optimal time and optimal time concentration of the GNP therapeutic nanomedicine effect for further determination. This assay is also used to obtain IC 50 values and the value of lethal doses of GNP therapeutic nanopharmaceuticals. The average cell viability values, neutral red assay, absorbance at different concentrations and percentage viability are summarized in figure 9, and compared statistically (n-3), the lethal dose 50(IC 50) for GNP is 10 μ l (50 μ g) for nanopharmaceuticals and 50 μ l for lethal dose. Fig. 10(a) shows that the different concentrations and increasing drug effect concentrations show a decrease in cell viability, as shown in fig. 10(b), accumulated at the two time points 6 and 24 hours. Mean and variance analysis and data lie within a confidence interval of p < 0.0001.
Cytotoxicity and drug binding at different times, cytotoxicity was observed at different time points according to cell viability and live-dead cell comparison, as shown in figure 11. After each time point, the plates were viewed upside down. Cell morphology and drug fluorescence were observed with a fluorescence microscope. Cells began to show a change in cell morphology in the test medium compared to the control, and the same pattern was that the last time point was observed. The cells appeared round, wrinkled, pale-bordered, indicating that the drug was present only on the cells and not in the blank. After 6 hours, fluorescence observation confirmed the presence of the nanomedicine, particularly doxorubicin, in the cells indicating its presence in the nucleus, as shown in fig. 12. After 6 hours, the cells began to clamp and showed GNPs and doxorubicin concentration in the cells as shown in fig. 13. The fluorescence enhancement dots and magnified images after 12 hours show the separation of the nanoparticles from the doxorubicin. Nanoparticles were observed in the cytoplasm, while most doxorubicin was in the nucleus, as shown in fig. 14. After 24 hours, the cells were killed with doxorubicin and then released from the cells. At this point, as shown in fig. 15, the cells began to lose fluorescence and at 72 hours the cells appeared to darken under the fluorescence microscope. Cytotoxicity studies have shown that GNPs have therapeutic effect nano-drugs are effective even at 72 hours due to the slow release of doxorubicin. Feasibility studies show different effects of the drug at different times. The drug was most effective within 12 hours and 24 hours. The viability was only 83% at 6 hours and 43% at 12 hours. Comparison of mean cell viability, as shown in fig. 16(a), the different time points indicated that the data were within the effective region (p < 0.0001). Live and dead cells were counted and their mean was compared by ANOVA. Studies showed that the data lie within the valid interval, with p < 0.0001. Comparison of live and dead cells also confirmed that the feasibility study for a potent drug effect remained 72 hours after 12 hours, as shown in fig. 16 (b).
Intracellular GNP and doxorubicin tracking and fluorescence intensity measurements were performed at different time points, and fluorescence intensity measurements at different time points were subtracted using Image J software to subtract the anti-fluorescence and study the independence of GNP and doxorubicin fluorescence. Subtracting the red from the green and going from the red to the green fluorescence plot, GNP and doxorubicin were traced inside the cell as shown in fig. 17, which shows receptor binding and intracellular localization of GNP nmr nano-drugs with binding and fluorescence intensity changes correlated with nanoparticle and doxorubicin concentrations at 6, 12, 24, 48, 72 hours. Most of the nano-drugs bound to the cells after 6 hours and showed fluorescence on the whole cells. After 12 hours, the cells emitted clear combined fluorescence, but after subtraction of the anti-fluorescence, the decrease indicated that most doxorubicin was still associated with GNP. Doxorubicin is therefore traced not only in the nucleus, but also in the cytoplasm with GNPs after 12 hours. Within 24 and 48 hours, most of the doxorubicin was localized in the nucleus and GNPs in the cytoplasm, and within 72 hours, the intracellular concentration began to disappear. The green fluorescence intensity is in direct proportion to the amount of GNP nanoparticles, and the red fluorescence intensity is proportional to the amount of doxorubicin. Twenty cells per group were used to calculate intensities and compare their means by analysis of variance as shown in figure 18. As shown in fig. 19, the points showing higher intensity were 6 hours, and the time at which the intensity started to decrease after 24 hours was 12 hours, respectively. Data were located at time intervals of p < 0.0001.
GNP tumor therapy nanomedicines bind to receptors on gastric cancer tissue (in vitro), and frozen sections of gastric tissue are incubated with GNP nanomedicines to examine the binding capacity of the nanomedicines to target tissue in vitro. As shown in fig. 20, successful binding of GNP tumor treating nano-drugs to transferrin receptor on gastric tissue was demonstrated. Fluorescent blue and green filters showed the presence of GNP nanoparticles in the tissue, while red fluorescence confirmed doxorubicin in the tissue. GNP tumor treatment nano-drugs showed successful and robust binding to transferrin receptor on gastric cancer tissues. The combination also proves that the preparation does not influence the combination of transferrin and transferrin receptor, and can be used for in vivo examination of the curative effect of the medicine.
Therapeutic efficacy (in vivo) of GNP therapeutic Nanoparticulates
GNP treatment the therapeutic effect and intracellular drug management were examined by oral administration of nano-drugs equivalent to 5mg/kg doxorubicin.
Receptor binding, intracellular tracking, drug release and fluorescence intensity at different time points, immunohistochemical study of nano-therapeutic drug oral dose in vitro tissue for receptor binding activity and intracellular tracking of gastric tissues collected after 3, 6, 12, 24, 48 and 72 hours of GNP. The nanomedicine binds to the receptor and begins to move the compartment inward into contact with the surface immediately thereafter. Figure 21 shows the binding and localization of the nanomedicine within the cell after 3, 6, and 12 hours. This was observed in tissue studies 12 hours after oral administration of the maximum drug. Doxorubicin fluoresces red and GNP fluoresces blue and green at different intensities. After 12 hours, the drug and nanoparticles began to clear from the cells, and fluorescence intensity decreased at 48 and 72 hours. As shown in fig. 22, the fluorescence intensity in the tissue was measured at the above time points using image J software. Among three different fluorescence filters, at 67.17cm each2The intensities of the triplet were measured and their average values were statistically compared, and the results are shown in fig. 23. GNP tumor treatment nano-drugs show maximal binding to the receptor within 6 hours and start to clear from the tissue after their removal. After 48 hours, most of the drug was approximately 30-50nm red from the nanoparticle size (green fluorescence decreased), while nanoparticles with a size less than this were still shown by the blue emission filter intensity. However, gold nanoparticles of various sizes and doxorubicin showed the same pattern of tissue localization and removal from the tissue.
The effect of GNP therapeutic nano-drugs on body weight and survival rate of gastric cancer-treated mice, the anticancer efficacy of GNP tumor-treating nano-drugs on induced cancer mice (third group induced cancer) was examined. Three groups: the first saline, second doxorubicin, and third GNP therapeutic nano-drug treatment group was observed for 40 days. The experiment was scheduled for two months but terminated at 40 days due to death of the last mouse in the second group. In contrast to treatment with doxorubicin, saline and GNP nano-therapeutic drug treated mice were active all the time. Although the group was dizzy hours after the nano-drug was taken, it started to be active after a while. Mice in the second group of mice became lethargic and weak daily, while mice in the first group were still active. After a period of time, as shown in fig. 24 and 25, the survival rate of the first group began to decrease, 60% at the end of the study. The survival rate was zero at 40 days in the second group of mice, and the third group of mice showed 80% survival rate and possessed an active lifestyle. The effect of treatment on body weight is summarized in fig. 26, and the results show that the third group of mice, treated with GNP therapeutic nanomedicines, not only had an extended lifespan, but also were living healthily, and had a smooth weight gain.
ZnS QD nano therapeutic drug (in vitro)
Statistical comparisons (ANOVA) of the three ZnSQD nanocomplexes, lower concentrations of 5 μ l (5 μ g/μ l) (ZnS-Cys, ZnS-Cys-Dox and ZnS-Cys-Dox-transferrin) on colon cancer cell lines were performed to observe the therapeutic effects of the nanocomplexes, as shown in FIG. 27. ZnS-Cys has a toxic effect on cancer cells, but only 2% have 98% cell viability. ZnS-Cys-Dox is more toxic due to the binding of the chemotherapeutic drug doxorubicin and the cell viability is 85%. After transferrin binding, cell viability showed a difference of 81%. Data are within the confidence interval, indicating comparative significance of p < 0.0001.
After 6 and 24 hours of incubation, different concentrations of ZnS QD therapeutic nano-drug (equivalent to different concentrations of doxorubicin) were examined for colon cancer cells. The present invention was used to examine the optimum time and optimum time concentration for ZnS nanomedicine action for further assays. This assay was also used to obtain IC 50 values and the value of lethal dose ZnS QD for tumor treatment nano-drugs. Fig. 28 and 29 summarize the average viability percentages for different concentrations and make a statistical comparison (n-3). The lethal dose 50(IC 50) of ZnS QD nano-therapeutic drug was 15. mu.l (57. mu.g) and the lethal dose was 50. mu.l. The cumulative effect of different concentrations of drug at the two time points 6 and 24 hours indicates a decrease in cell viability. The mean was compared to ANOVA and data was placed with p-median <0.0001 at confidence intervals.
Cytotoxicity and drug binding of ZnS QD therapeutic nano-drugs at different time points cytotoxicity was observed at different time points as shown in figure 30 based on cell viability and comparison of live and dead cells. After each time point, the plates were viewed upside down. Cell morphology and drug fluorescence were observed with a fluorescence microscope. Cells initially showed a change in cell morphology in the test medium compared to the control, and the same pattern was observed until the last time point. The cellular appearance in the presence of ZnS QD nanotherapeutic drugs is different from the cellular morphology previously observed in GNP nanotherapeutic drug samples and from the control. The cell plate showed that the drug was only present on the cells and not in the blank. After 6 hours, fluorescence observation confirmed the presence of the nanomedicine, particularly doxorubicin, in the cells indicating its presence in the nucleus, as shown in fig. 31. After 6 hours, cells began to clamp and showed an increase in ZnS QD and doxorubicin concentrations. After the 12 hour time point, fluorescence increased as shown in fig. 32, and maximum fluorescence was observed at the 24 hour point. ZnS QD nanoparticles were observed in the cytoplasm, while most doxorubicin was in the nucleus, as shown in figure 33. After 24 hours, doxorubicin was used to kill and release from the cells. After 48 hours the nuclei were quiescently filled with doxorubicin as shown in FIG. 34. At 72 hours the cells died and the cells began to discolour and deform. Cytotoxicity studies showed that ZnS QD treatment of nanomedicines was effective even within 72 hours. Feasibility studies confirmed that previously observed nano-drugs affecting GNP tumor therapy showed the greatest efficacy within 12 hours. As shown in fig. 35, the ZnS QD therapeutic nano-drug continued to have a severe effect on cells after 12 hours until the last examination time point was 72 hours. The survival rate at 6 hours was only 84% and at 12 hours 44%, which was still close to this level at 24 hours and 48 hours. Statistical comparison of cell viability averages at different time points indicated that the data were located in the important region (p < 0.0001). Live and dead cells were counted and their mean values were compared by analysis of variance. Studies indicate that data exists between significant regions of p < 0.0001. Live and dead cell comparison also verifies that viability studies indicate that the ZnS QD therapeutic nano-drug has a prolonged effect on cells.
Intracellular ZnS quantum dots and doxorubicin tracking and fluorescence intensity measurements at different time points, back-fluorescence was subtracted at different time points using Image J software, and ZnS QD and doxorubicin fluorescence were studied independently. The red was subtracted from the green and shifted from the red to the green fluorescence plot, and ZnS QD and doxorubicin were traced inside the cells as shown in figure 36. Most of the nano-drugs were incorporated in the cells after 6 hours and showed fluorescence throughout the cells. At 12 hours, the cells fluoresced clearly combined, but the decrease after subtraction of the back-fluorescence indicates that most doxorubicin was still present with ZnS QDs. Therefore ZnS QD traced doxorubicin not only in the nucleus, but also in the cytoplasm at 12 hours. After 24 and 48 hours, most of the doxorubicin was localized in the nucleus, while ZnS QD was localized in the cytoplasm, and after 72 hours, the intracellular concentration began to disappear. The green fluorescence intensity is proportional to the number of ZnS QD nanoparticles, and the red fluorescence intensity is proportional to the amount of doxorubicin. After subtraction, the fluorescence intensity anti-fluorescence was analyzed by image J. Twenty cells per group were used to calculate intensities and their means compared by analysis of variance are shown in figure 37. Fig. 38 shows that the higher intensity points were 6 hours and 12 hours, respectively, and the intensity started to decrease after 24 hours. Data are in the significant interval with p < 0.0001.
ZnS QD therapeutic nano-drugs bind to receptors on gastric cancer tissues (in vitro), and frozen sections of gastric tissues are incubated with ZnS QD nano-therapeutic drugs to examine the ability of the nano-drugs to bind to target tissues in vitro. Figure 39 shows successful binding of ZnS QD tumor therapy nanomedicine to transferrin receptor on gastric tissue. Fluorescence in blue and green filters indicated the presence of ZnS QD nanoparticles in the tissue, while red fluorescence confirmed the presence of doxorubicin in the tissue. In vitro studies confirmed that ZnS QD tumor treatment nanomedicines do not affect the receptor binding site of transferrin. The transferrin in the therapeutic nano-drug is completely activated and can be combined with transferrin to a receptor.
Therapeutic efficacy (in vivo) of ZnS QD therapeutic Nanoparticulates
ZnS QD nano therapeutic drug equivalent to 5mg/kg doxorubicin was orally administered to examine the therapeutic effect and intracellular drug treatment method. The results of ZnS QD therapeutic nanomedicines were used to confirm and support the behavior and effects of GNP therapeutic nanomedicines.
Receptor binding, intracellular tracking, drug release and fluorescence intensity of ZnS QD therapeutic nanomedicines at different time points, receptor binding activity of tissues and intracellular cell tracking were observed by ex vivo immunohistochemical studies of gastric tissues collected 3, 6, 12, 24, 48 and 48 hours after oral administration of ZnS QD nanomedicines. The nanomedicine receptor binding and localization pattern mimics the GNP therapeutic nanomedicine pattern with no change in pattern. FIG. 40 shows binding and localization of intracellular nano-drugs after 3, 6 and 12 hours. The largest drug was observed in the tissue study after 12 hours, and also after 12 hours in the GNP therapeutic nano-drug study. The decrease in fluorescence was observed by confirming the gradual elimination behavior after 12 hours. The size range of ZnS QD nanoparticles is lower than that of GNP therapeutic nanomedicines, so in the case of quantum dots, blue fluorescence is clearer and more intense than green fluorescence. The fluorescence intensity was measured in a 67.17cm2 area with image J and compared using SPSS 26, as shown in FIG. 41. The results are summarized in fig. 42, ZnS QD therapeutic nanomedicine showed maximum binding to the receptor at 6 hours and gradually started to leave the tissue. After 48 hours, most of the doxorubicin was cleared from the tissue as shown by the decrease in red fluorescence. ZnS QDs are smaller in size than GNPs, but they escape from tissue more efficiently at the 72 hour time point.
The influence of ZnS QD nano therapeutic drugs on mouse body weight and survival rate during gastric cancer treatment was also examined for the anticancer effect of ZnS QD therapeutic nano drugs, three groups: the first group of saline, the second group of doxorubicin and the third group of GNP therapeutic nano-drugs re-experiment was terminated at day 40 because the survival rate of the group 2 mice at day 40 was zero. Throughout the treatment, the group treated with ZnS QD therapeutic nanomedicine remained active compared to the doxorubicin group. As shown in fig. 43 and 44, the survival rate of ZnS QD-treated group was 80%, but the first mice died after 18 days, rather than the 15-day group observed in GNP animal treatment. The effect of treatment on body weight was observed in the first 15 days and the results are shown in fig. 45 and 46, where ZnS QD treatment nano-drugs not only can prolong life but also have a healthy life and have a smooth weight gain pattern.
Example 4 pharmacokinetic and biodistribution Studies
Pharmacokinetics (in vitro) of GNP tumor treatment nanomedicines
The pH-dependent drug release of GNP therapeutic nano-drugs was performed at acidic pH5 and neutral pH7.5 (in vitro). The purpose of the experiment was to examine the kinetic release behavior conditions of the drug at different pH. Acidic conditions are prevalent in the vicinity of tumors and in the stomach, while the physiological pH is almost 7.5 for plasma and organs. The pH of the medium containing the nano-drug was also 7.5, and therefore, the drug release behavior was examined to see the stability of the drug and the release at the desired site. The drug release behavior of GNP tumor treatment nano-drugs at pH5 and pH7.5 is shown in fig. 47 and 48. Drug release behavior demonstrates the stability of the therapeutic nano-drug at pH7.5 and dominant release behavior at acidic pH 5. The drug release behavior indicates that doxorubicin is released, the free form and the drug release rate are greater at pH5, and the drug is stable at pH 7.5. These results also predict the in vivo stability of the drug and its release at the tumor site and in the stomach due to pH-dependent release. At pH5, half of the doxorubicin was released in 85 hours, as shown in fig. 49, while at pH7.5, it took 850 minutes to release half of the conjugated doxorubicin, as shown in fig. 50. Data points are presented as mean values of three in figure 49 and figure 50 as ± SD.
The GNP nano-therapeutic drug released doxorubicin in cell culture, and doxorubicin release was measured in two sets of reactions in cell culture. Measurements were made between the first target concentration and the non-target concentration, and at the second time, the concentration of doxorubicin in dead cells and the culture medium was determined by HPLC. FIG. 51 shows chromatograms of pure doxorubicin (a), standard doxorubicin (b), doxorubicin (c) extracted from dead cells and culture medium, and doxorubicin (d) extracted from plasma.
Doxorubicin release from targeted and non-targeted GNPs as shown in figure 52, the nanocomposite formulation, doxorubicin concentration in dead cells incubated with GNP-Cys-Dox and GNP-Cys-Dox, was calculated at 6 hours and 24 hours for reaction with GNP-Cys-Dox-Trans, values compared by analysis of variance, with p <0.005 for the non-target population over 24 hours. Figure 53 shows that the targeted GNP nanocomplexes showed higher doxorubicin concentrations at 24 hours than the non-targeted GNP nanocomplexes.
The doxorubicin concentration in GNP tumor treatment nano-drugs was analyzed by HPLC at different time points, under different conditions in media with dead cells and compared by analysis of variance. The maximum concentration of doxorubicin was found to be 72 hours, then 12 hours. As shown in fig. 54, the lowest concentration was recorded at 6 hours. All reactions were performed in triplicate and data expressed as standard deviation and significance (p <0.005) as mean values, as shown in FIG. 55.
Pharmacokinetics (in vivo) of GNP tumor treatment nanomedicines
The time profile of blood activity of GNP therapeutic nano-drug release doxorubicin was studied in two groups. One group received the nano-drug via the intravenous route (IV route) and the other group via the oral route.
Pharmacokinetics of GNP nmr nano-drugs, intravenous administration, and following intravenous administration of GNP therapeutic nano-drugs, drug distribution in plasma was characterized by HPLC analysis. Figure 56 summarizes the data collected for HPLC analysis of doxorubicin release in plasma at different time points and for statistical comparison (analysis of variance). The analysis showed that the data are within a significant time interval (p)<0.001) and all pharmacokinetic parameters were calculated and are summarized in figure 57. The clearance rate of GNP-Cys-Dox-transferrin is 0.002L/h, and the plasma half-life period (t)1/2) It was 12.70 hours. Over time, plasma drug concentration decreased for the time and small amounts of doxorubicin were observed at 48 and 72 hours.
Plasma concentration management of doxorubicin was analyzed by HPLC at various times after oral administration and the data shown in fig. 58 are presented as mean values with standard deviation (n-3) and as a significant interval with p < 0.001. The elevated drug concentration in plasma shown in figure 59 was administered orally and reached a maximum concentration at 48 hours. After 48 hours to 72 hours, the concentration line showed a sharp drop and eliminated the drug from the plasma. Figure 60 summarizes pharmacokinetic parameters after oral dosing.
Biodistribution of GNP tumor treatment nanomedicines, and also after detection of doxorubicin concentrations in liver, heart, kidney and stomach tumors with 485 excitation and 525 emission fluorescence scanners for IV and PO dosing (luciferase reader).
Figure 61 shows doxorubicin concentration and PO dose in the liver at different time points after intravenous infusion. The results shown in fig. 62 demonstrate that doxorubicin decreased in liver tissue over time. The doxorubicin concentration in the liver tissue after oral administration was less than the increase in blood concentration after intravenous infusion at all time points and showed rapid clearance from the liver tissue.
Figure 63 summarizes doxorubicin concentrations and PO doses in the heart at various time points after intravenous infusion. Data represent mean (n-3) with standard deviation, significance (p <0.005) was calculated by ANOVA and independent T-test. Figure 64 shows doxorubicin concentrations at various time points, demonstrating rapid elimination of doxorubicin from the heart, with orally administered doxorubicin concentrations lower than intravenous injection.
FIG. 65 shows the elimination of doxorubicin from the kidney at various time points, with the mean (n-3) representing the deviation and significance (p < 0.05). Doxorubicin concentration decreased in time and showed rapid elimination from the body, as shown in fig. 66. Most doxorubicin is found in the kidney and cleared from the body after oral administration.
Biodistribution experiments were performed in mice that induced gastric cancer (group 3). Post-intravenous administration, only a small amount could reach the target site, but still rarely showed an effect until 48 hours. FIG. 67 shows that most drugs administered orally bind to the target site and show higher concentrations in the gastric tumor fraction at 48 hours, and the data in FIG. 68 are summarized as mean (n-3) with standard deviation and significance (p < 0.0001).
Pharmacokinetics (in vitro) of ZnS QD therapeutic Nanoparticulates
pH-dependent drug release of ZnS QD therapeutic nano-drug the drug release behavior of ZnS QD therapeutic nano-drug at pH5 and pH7.5 is as shown in fig. 69 and fig. 70. The drug release behavior confirmed the stability of the nano-drug at pH7.5 and the significant release behavior at acidic pH 5. Figure 69 is plotted as percent drug release at different time intervals. Doxorubicin was released in the medium at pH5 at a higher release rate at pH7.5 than at pH 7.5. These results reinforce the previously observed release pattern of doxorubicin from GNP tumor treating nano-drugs. In both drugs doxorubicin was conjugated to the carboxyl group of cysteine and thus released in a similar manner. At pH5, half of the doxorubicin was released in 140 hours, while at pH7.5, half of the bound doxorubicin took 800 minutes to be released. The data points are shown in fig. 71 and 72.
Doxorubicin release from ZnS QD therapeutic nano-drug, nano-drug in cell culture (in vitro) in cell culture, doxorubicin release was measured in two sets of reactions. Dead cells and media were measured by HPLC at different time points at the first target and non-target concentrations and at the second doxorubicin concentration.
Doxorubicin release from targeted and non-targeted ZnS quantum dots, therapeutic nanocomposites were calculated at 6 and 24 hours for doxorubicin concentration and ZnS-Cys-Dox-transferrin reaction time in responder dead cells incubated with ZnS-Cys-Dox and ZnS, respectively. The values were compared by analysis of variance as shown in fig. 73. Meaning that the 24 hour localization to the non-localized group p < 0.01. The targeted ZnS QD nanocomposite showed higher doxorubicin concentrations, as shown in figure 74, over the untargeted ZnS QD nanocomposite in 24 hours.
The concentration of doxorubicin in ZnS QD was analyzed by HPLC for the concentration of doxorubicin in media with dead cells at different time points and compared by ANOVA. The maximum concentration of doxorubicin was found to be 72 hours, then 12 hours. After 12 hours, the next two time points remained almost stable as shown in fig. 75. All reactions were performed in triplicate and data expressed as mean values. Standard deviation and significance (p <0.005) as shown in figure 76.
ZnS QD tumor treatment NanoTaharmaceutical pharmacokinetics (in vivo)
The temporal profile of doxorubicin blood activity of the ZnS QD therapeutic nano-drug was divided into two groups of studies. One group received the nano-drug via intravenous route and the other group via oral route. The biodistribution of doxorubicin was also characterized after intravenous and oral administration.
Following intravenous injection of ZnS QD, plasma drug profiles were characterized by HPLC analysis. Figure 77 summarizes data collected from HPLC of doxorubicin in plasma at different time points and performs statistical comparisons (ANOVA). Analysis showed data at significant time intervals (p <0.001) as shown in FIG. 78. FIG. 79 shows that the plasma half-life of ZnS-Cys-Dox-transferrin is 16.91h at a rate of 0.001L/h. Over time, the drug concentration in plasma decreased with small amounts of doxorubicin observed at 48 and 72 hours.
Pharmacokinetics of ZnS QD therapeutic nanomedicines after oral administration, plasma concentrations of doxorubicin were analyzed by HPLC at various times after oral administration. Data are shown in fig. 80 as mean values with standard deviation (n-3) and intervals with p <0.001 as the significand. The drug concentration in the plasma increased after oral administration, and the drug was administered to reach the maximum concentration in 48 hours. After 48 to 72 hours, the concentration lines showed a sharp drop and elimination of drug from the plasma, fig. 81. Figure 82 summarizes pharmacokinetic parameters after oral dosing.
Biodistribution of nano-drugs for ZnS QD tumor therapy, and IV and PO doses (luciferase labeling) with 485 excitation and 525 emission by a fluorescence scanner for postoperative detection of doxorubicin concentrations in liver, heart, kidney and stomach tumors.
Figure 83 shows doxorubicin concentration and PO dose in the liver at different time points after intravenous injection. As shown in fig. 84, the results confirmed the reduction of doxorubicin in liver tissue over time. The doxorubicin concentration in the liver tissue after oral administration was less than the blood concentration after intravenous injection at all time points and showed rapid clearance from the liver tissue.
Figure 85 summarizes doxorubicin concentrations and PO doses in the heart at various time points after intravenous injection. Data have mean values of standard deviation (n-3), significance (p <0.005) calculated by ANOVA and independent T-test. Figure 86 shows doxorubicin concentrations at different time points. The doxorubicin concentrations tracked in the heart were extremely low, and the results confirmed the rapid elimination of doxorubicin from the heart. Oral administration of doxorubicin is less concentrated than intravenous injection.
FIG. 87 shows the elimination of doxorubicin from the kidney at various time points and the data obtained from the analysis represent the deviation and significance (p <0.001) as the mean value of the standard (n-3). Doxorubicin concentrations decreased over time, with no significant doxorubicin concentrations after intravenous and oral administration at the 6 hour time point, as shown in fig. 88. The kidney was orally taken at higher concentrations for 24 hours, but most doxorubicin was cleared from the body at 48 hours.
Biodistribution experiments were performed in mice that induced gastric cancer (group 3). Post-intravenous administration, only small amounts of doxorubicin could reach the target site, but at 48 hours, very small amounts were also tracked in the stomach. Oral administration showed that most of the drug bound to the target site, with a redistribution pattern 89 after 48 hours at higher concentrations in gastric tumors. The data in figure 90 are summarized as mean (n-3) with standard deviation and significance (p < 0.0001).
Claims (7)
1. The preparation method of the nano-targeted drug and the application of the nano-targeted drug in the treatment of gastric cancer are characterized in that the nano-targeted drug comprises GNP therapeutic nano-drug and ZnS QD therapeutic nano-drug.
2. The preparation method of the nano-targeting drug and the application of the nano-targeting drug in the treatment of gastric cancer according to claim 1, wherein the linker of the GNP therapeutic nano-drug is cysteine, the targeting ligand is transferrin, the chemotherapeutic drug is doxorubicin, and the nano-carrier is GNP.
3. The preparation method of the nano-targeted drug and the application thereof in the treatment of gastric cancer according to claim 1, wherein the linker of the ZnS QD therapeutic nano-drug is cysteine, the targeting ligand is transferrin, the chemotherapeutic drug is doxorubicin, and the nano-carrier is ZnS QD.
4. The preparation method of the nano-targeted drug and the application of the nano-targeted drug in the treatment of gastric cancer according to claim 1, wherein the preparation method of the nano-targeted drug comprises the preparation of GNP therapeutic nano-drugs and the preparation of ZnS QD therapeutic nano-drugs.
5. The method for preparing nano-targeted medicament according to claim 4, wherein the preparation of GNP therapeutic nano-medicament comprises the synthesis of GNP, the combination of cysteine and GNP, the combination of adriamycin and GNP-Cys, and the combination of transferrin and GNP-Cys-Dox.
6. The method for preparing nano targeted drug according to claim 4, characterized in that the preparation of ZnS QD therapeutic nano drug comprises the synthesis of ZnS QD quantum dots coated with cysteine, the combination of adriamycin and ZnS QD-Cys, and the combination of transferrin and ZnS QD-Cys-Dox.
7. The preparation method of the nano-targeted drug and the application of the nano-targeted drug in the treatment of gastric cancer according to claim 1, wherein cysteine-coated GNP and transferrin are conjugated to serve as a targeted diagnostic probe.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011525066.4A CN112494660B (en) | 2020-12-22 | 2020-12-22 | Preparation method of nano targeting drug and application of nano targeting drug in treating gastric cancer |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011525066.4A CN112494660B (en) | 2020-12-22 | 2020-12-22 | Preparation method of nano targeting drug and application of nano targeting drug in treating gastric cancer |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112494660A true CN112494660A (en) | 2021-03-16 |
CN112494660B CN112494660B (en) | 2023-11-21 |
Family
ID=74923029
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011525066.4A Active CN112494660B (en) | 2020-12-22 | 2020-12-22 | Preparation method of nano targeting drug and application of nano targeting drug in treating gastric cancer |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112494660B (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2022198438A1 (en) * | 2021-03-23 | 2022-09-29 | Wuhan Vast Conduct Science Foundation Co., Ltd. | Ligand-bound zinc sulfide nanoparticles, methods for making the same, and their use for treatment |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102349999A (en) * | 2011-08-23 | 2012-02-15 | 吉林大学 | Multifunctional adriamycin precursor medicament as well as preparation method and application thereof |
CN110178028A (en) * | 2017-01-09 | 2019-08-27 | 密苏里大学管理机构 | Targeting peptide-doxorubicin-gold nano conjugate for oncotherapy |
-
2020
- 2020-12-22 CN CN202011525066.4A patent/CN112494660B/en active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102349999A (en) * | 2011-08-23 | 2012-02-15 | 吉林大学 | Multifunctional adriamycin precursor medicament as well as preparation method and application thereof |
CN110178028A (en) * | 2017-01-09 | 2019-08-27 | 密苏里大学管理机构 | Targeting peptide-doxorubicin-gold nano conjugate for oncotherapy |
Non-Patent Citations (9)
Title |
---|
MADEEHA SHAHZAD LODHI ET AL.: "Engineering Quantum Dot (Cadmium Sulfide) on Antibodies for Fluoroimmunoassays", 《JOURNAL OF NANOMATERIALS》 * |
MADEEHA SHAHZAD LODHI ET AL.: "Purification of transferrin by magneticnanoparticles and conjugation with cysteinecapped gold nanoparticles for targeting diagnosticprobes", 《PREPARATIVE BIOCHEMISTRY AND BIOTECHNOLOGY》 * |
SUNIL KUMAR ET AL.: "Functionalization and characterization of ZnS quantum dots using", 《JOURNAL OF MATERIALS SCIENCE:MATERIALS IN ELECTRONICS》 * |
VIVEK PANDEY ET AL: "Nucleation temperature-controlled synthesis and in vitro toxicity evaluation of L-cysteine capped Mn:ZnS quantum dots for intracellular imaging", 《LUMINESCENCE》 * |
王晓波: "《药物运释系统》", 31 August 2007, 中国医药科技出版社 * |
葛金龙: "《材料化学专业实验》", 30 September 2019 * |
金征宇等: "《基因与纳米探针:医学分子成像理论与时间》", 30 November 2017, 天津医药科学技术出版社 * |
韩晓军: "《生物功能化界面》", 31 January 2017, 哈尔滨工业大学出版社 * |
顾月清 等: "《生物医学工程技术》", 30 June 2017, 中国医药科技出版社 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2022198438A1 (en) * | 2021-03-23 | 2022-09-29 | Wuhan Vast Conduct Science Foundation Co., Ltd. | Ligand-bound zinc sulfide nanoparticles, methods for making the same, and their use for treatment |
Also Published As
Publication number | Publication date |
---|---|
CN112494660B (en) | 2023-11-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Fan et al. | Ultrasmall gold nanoparticles in cancer diagnosis and therapy | |
US11497809B2 (en) | Multifunctional nanoparticle compositions and uses thereof | |
US9486480B2 (en) | Surface-modified heavy metal nanoparticles, compositions and uses thereof | |
KR102081666B1 (en) | Phamaceutical composition for treating cancer | |
Zhang et al. | pH-driven targeting nanoprobe with dual-responsive drug release for persistent luminescence imaging and chemotherapy of tumor | |
US10646472B2 (en) | Composition comprising a photosensitive compound in a polymeric nanoparticle, and a method of using the composition | |
US20130315834A1 (en) | Nanoprobe comprising gold colloid nanoparticles for multimodality optical imaging of cancer and targeted drug delivery for cancer | |
US9132098B2 (en) | Stable nanocomposition comprising doxorubicin, process for the preparation thereof, its use and pharmaceutical compositions containing it | |
Dash et al. | Hyaluronic acid-modified, IR780-conjugated and doxorubicin-loaded reduced graphene oxide for targeted cancer chemo/photothermal/photodynamic therapy | |
Cao et al. | A triple modality BSA-coated dendritic nanoplatform for NIR imaging, enhanced tumor penetration and anticancer therapy | |
Zhao et al. | Multifunctional magnetic nanoparticles for simultaneous cancer near-infrared imaging and targeting photodynamic therapy | |
US9675714B1 (en) | Graphene based theranostics for tumor targeted drug/gene delivery and imaging | |
Wu et al. | Pea protein/gold nanocluster/indocyanine green ternary hybrid for near-infrared fluorescence/computed tomography dual-modal imaging and synergistic photodynamic/photothermal therapy | |
US10201622B2 (en) | Tumour-targeted theranostic | |
Lee et al. | Gold-stabilized carboxymethyl dextran nanoparticles for image-guided photodynamic therapy of cancer | |
Shabani et al. | The brilliance of nanoscience over cancer therapy: Novel promising nanotechnology-based methods for eradicating glioblastoma | |
Lugani et al. | Dual Immunostimulatory Pathway Agonism through a Synthetic Nanocarrier Triggers Robust Anti‐Tumor Immunity in Murine Glioblastoma | |
Zhang et al. | Current advances in the application of nanomedicine in bladder cancer | |
CN112494660B (en) | Preparation method of nano targeting drug and application of nano targeting drug in treating gastric cancer | |
Foo et al. | Unravelling the potential of graphene in glioblastoma therapy | |
Gholami et al. | A comprehensive perspective of trastuzumab-based delivery systems for breast cancer treatment | |
CN114848843B (en) | Chemotherapeutic synergistic targeting combined treatment nano-drug and application thereof in tumor treatment | |
Hu et al. | Challenges and advances for glioma therapy based on inorganic nanoparticles | |
Zhang et al. | Multifunctional nanocarriers based on graphitic-C 3 N 4 quantum dots for tumor-targeted, traceable and pH-responsive drug delivery | |
US20220323432A1 (en) | Nanoparticles for delivering drug, whose surface is modified with peptide for targeting brain cancer, preparation method thereof and use thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |