CN113209020B - 一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用 - Google Patents
一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用 Download PDFInfo
- Publication number
- CN113209020B CN113209020B CN202110540998.4A CN202110540998A CN113209020B CN 113209020 B CN113209020 B CN 113209020B CN 202110540998 A CN202110540998 A CN 202110540998A CN 113209020 B CN113209020 B CN 113209020B
- Authority
- CN
- China
- Prior art keywords
- aiegenes
- mpa
- enzyme
- diagnosis
- aiegens
- 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.)
- Active
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 79
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 56
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 54
- 239000000693 micelle Substances 0.000 title claims abstract description 47
- 238000003745 diagnosis Methods 0.000 title claims abstract description 33
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 23
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 22
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000002246 antineoplastic agent Substances 0.000 title abstract description 7
- 229940041181 antineoplastic drug Drugs 0.000 title abstract description 7
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 14
- 239000003504 photosensitizing agent Substances 0.000 claims abstract description 14
- 239000007864 aqueous solution Substances 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 13
- ZNWYPDKJWGLSQU-UHFFFAOYSA-N 2-[(4-bromo-2,1,3-benzothiadiazol-7-yl)methylidene]propanedinitrile Chemical compound BrC1=CC=C(C=C(C#N)C#N)C2=NSN=C12 ZNWYPDKJWGLSQU-UHFFFAOYSA-N 0.000 claims abstract description 6
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000006069 Suzuki reaction reaction Methods 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 36
- 239000000032 diagnostic agent Substances 0.000 claims description 16
- 229940039227 diagnostic agent Drugs 0.000 claims description 16
- 150000003384 small molecules Chemical class 0.000 claims description 16
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 15
- 206010028980 Neoplasm Diseases 0.000 claims description 13
- 239000003814 drug Substances 0.000 claims description 13
- 238000000502 dialysis Methods 0.000 claims description 12
- 238000003786 synthesis reaction Methods 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 11
- 239000000843 powder Substances 0.000 claims description 8
- 239000003960 organic solvent Substances 0.000 claims description 6
- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 claims description 6
- 229940124597 therapeutic agent Drugs 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 4
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 4
- 239000012498 ultrapure water Substances 0.000 claims description 4
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 3
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 3
- 201000010881 cervical cancer Diseases 0.000 claims description 3
- 238000001308 synthesis method Methods 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- 206010060862 Prostate cancer Diseases 0.000 claims description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 2
- 238000006555 catalytic reaction Methods 0.000 claims description 2
- 230000000259 anti-tumor effect Effects 0.000 claims 1
- 238000005286 illumination Methods 0.000 abstract description 15
- 210000004881 tumor cell Anatomy 0.000 abstract description 13
- 238000001338 self-assembly Methods 0.000 abstract description 9
- 230000006870 function Effects 0.000 abstract description 7
- 230000000638 stimulation Effects 0.000 abstract description 7
- 102000005600 Cathepsins Human genes 0.000 abstract description 5
- 108010084457 Cathepsins Proteins 0.000 abstract description 5
- 238000010532 solid phase synthesis reaction Methods 0.000 abstract description 4
- 230000004043 responsiveness Effects 0.000 abstract description 3
- 230000006698 induction Effects 0.000 abstract description 2
- 238000005580 one pot reaction Methods 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 230000002194 synthesizing effect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 39
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 18
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 17
- 239000003642 reactive oxygen metabolite Substances 0.000 description 17
- 239000011347 resin Substances 0.000 description 17
- 229920005989 resin Polymers 0.000 description 17
- 150000001413 amino acids Chemical class 0.000 description 11
- 238000006482 condensation reaction Methods 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 9
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 9
- 238000002428 photodynamic therapy Methods 0.000 description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 8
- 150000001408 amides Chemical class 0.000 description 8
- 231100000263 cytotoxicity test Toxicity 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 238000003384 imaging method Methods 0.000 description 6
- BBEAQIROQSPTKN-UHFFFAOYSA-N pyrene Chemical compound C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 BBEAQIROQSPTKN-UHFFFAOYSA-N 0.000 description 6
- 238000006467 substitution reaction Methods 0.000 description 6
- 230000008685 targeting Effects 0.000 description 6
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 5
- 230000002776 aggregation Effects 0.000 description 5
- 238000004220 aggregation Methods 0.000 description 5
- 231100000135 cytotoxicity Toxicity 0.000 description 5
- 230000003013 cytotoxicity Effects 0.000 description 5
- 230000005284 excitation Effects 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- XDFNWJDGWJVGGN-UHFFFAOYSA-N 2-(2,7-dichloro-3,6-dihydroxy-9h-xanthen-9-yl)benzoic acid Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC(Cl)=C(O)C=C2OC2=CC(O)=C(Cl)C=C21 XDFNWJDGWJVGGN-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 4
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 4
- 206010063344 microscopic polyangiitis Diseases 0.000 description 4
- 238000011056 performance test Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 230000007704 transition Effects 0.000 description 4
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 3
- 239000005964 Acibenzolar-S-methyl Substances 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 108010087230 Sincalide Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000010609 cell counting kit-8 assay Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 230000012202 endocytosis Effects 0.000 description 3
- GVEPBJHOBDJJJI-UHFFFAOYSA-N fluoranthrene Natural products C1=CC(C2=CC=CC=C22)=C3C2=CC=CC3=C1 GVEPBJHOBDJJJI-UHFFFAOYSA-N 0.000 description 3
- 230000010354 integration Effects 0.000 description 3
- 238000001254 matrix assisted laser desorption--ionisation time-of-flight mass spectrum Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 230000000149 penetrating effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 description 2
- FNQJDLTXOVEEFB-UHFFFAOYSA-N 1,2,3-benzothiadiazole Chemical compound C1=CC=C2SN=NC2=C1 FNQJDLTXOVEEFB-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 102000004225 Cathepsin B Human genes 0.000 description 2
- 108090000712 Cathepsin B Proteins 0.000 description 2
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 description 2
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 230000005281 excited state Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000001678 irradiating effect Effects 0.000 description 2
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 2
- 231100000956 nontoxicity Toxicity 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 231100000820 toxicity test Toxicity 0.000 description 2
- ODHXBMXNKOYIBV-UHFFFAOYSA-N triphenylamine Chemical compound C1=CC=CC=C1N(C=1C=CC=CC=1)C1=CC=CC=C1 ODHXBMXNKOYIBV-UHFFFAOYSA-N 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- FODJWPHPWBKDON-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-[(2-methylpropan-2-yl)oxy]-4-oxobutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 FODJWPHPWBKDON-IBGZPJMESA-N 0.000 description 1
- UMRUUWFGLGNQLI-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UMRUUWFGLGNQLI-QFIPXVFZSA-N 0.000 description 1
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
- JFLSOKIMYBSASW-UHFFFAOYSA-N 1-chloro-2-[chloro(diphenyl)methyl]benzene Chemical compound ClC1=CC=CC=C1C(Cl)(C=1C=CC=CC=1)C1=CC=CC=C1 JFLSOKIMYBSASW-UHFFFAOYSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 239000012223 aqueous fraction Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 229920000891 common polymer Polymers 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000011258 core-shell material Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 125000004093 cyano group Chemical group *C#N 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 125000006575 electron-withdrawing group Chemical group 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 238000012632 fluorescent imaging Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000005283 ground state Effects 0.000 description 1
- -1 hexafluorophosphate Chemical compound 0.000 description 1
- 230000005525 hole transport Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000007777 multifunctional material Substances 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000012299 nitrogen atmosphere Substances 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 238000006552 photochemical reaction Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000001269 time-of-flight mass spectrometry Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 1
- 125000006617 triphenylamine group Chemical group 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- 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
- A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
- A61K49/0076—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion
- A61K49/0082—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion micelle, e.g. phospholipidic micelle and polymeric micelle
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D285/00—Heterocyclic compounds containing rings having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by groups C07D275/00 - C07D283/00
- C07D285/01—Five-membered rings
- C07D285/02—Thiadiazoles; Hydrogenated thiadiazoles
- C07D285/14—Thiadiazoles; Hydrogenated thiadiazoles condensed with carbocyclic rings or ring systems
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Inorganic Chemistry (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明属于抗肿瘤药物制备技术领域,具体公开了一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用。本发明以4,4'‑二甲氧基‑4''‑硼酸三苯胺和4‑溴‑7‑(2,2‑二氰基乙烯基)苯并[C][1,2,5]噻二唑为主要原料,采用Suzuki Coupling一步反应合成得到AIEgens‑1光敏剂;采用多肽固相合成法合成水溶性小分子功能肽;利用多肽本身的两亲性结构在水溶液中自组装形成胶束,进而实现对疏水性AIEgens‑1光敏剂的有效包埋,构建了一种酶敏感型多肽胶束型诊疗剂。本发明的酶敏感型多肽胶束型诊疗剂利用多肽本身的功能多样性,实现AIEgens‑1光敏剂在肿瘤细胞中的特异性富集,组织蛋白酶刺激响应性释放及光照刺激诱导的诊疗一体化功能。
Description
技术领域
本发明属于抗肿瘤药物制备技术领域,具体涉及一种酶敏感型多肽胶束型诊疗剂(AIEgens-1@MPA)及其在制备抗肿瘤药物中的应用。
背景技术
癌症的诊断治疗一体化将诊断和治疗合二为一,通过将药物和成像试剂集成于纳米载体中,有针对性地运输至肿瘤组织并对治疗效果进行实时监测,从而实现癌症早期诊断和精确治疗。目前,癌症治疗试剂的成功应用很大程度上依赖于多功能材料的设计和合理使用,开发高效低毒的诊疗药物对癌症的精准诊疗具有重大意义。
传统的有机荧光分子在高浓度溶液或聚集状态下往往会出现荧光猝灭现象(ACQ),因而在生物荧光成像等领域的应用受到了严重限制。聚集诱导发光分子(AIEgens)作为一种特殊的有机发光分子,在稀溶液中分子内部可以自由旋转和振动,导致能量的散失以非辐射跃迁为主,从而呈现极弱的荧光发射或非发射状态。但是在聚集态和固态中,AIEgens的分子内转动受到限制,能量散失从非辐射跃迁变为辐射跃迁,从而表现出强烈的荧光发射,从根本上解决有机荧光染料的ACQ问题。与此同时,AIEgens在特定波长的光照下吸收适当能量的光子后,处于基态(S0)的电子被激发至单重激发态(S1),然后通过系间窜跃到达三重激发态(T1)。处于T1的AIEgens与周围的氧分子反应产生形成自由基(O2 ·-和·OH)和氧化性物质(H2O2),或者直接与三线态氧(3O2)反应生成单线态氧(1O2)。这一类活性氧(ROS)具有很高的细胞毒性,对细胞膜、核酸及蛋白等生物活性物质会产生氧化损伤,进而加速细胞凋亡。这种利用光化学反应产生的ROS引起严重的细胞损伤的治疗手段称为光动力学治疗(PDT)。相较于传统的癌症治疗模式,PDT具有一系列优势,包括创伤小、时空选择性高、可控性强、毒副作用低和疗效高等优点,因而在肿瘤高效治疗方面具有巨大前景。这种将成像和治疗功能整合于同一种纳米材料,从而实现荧光成像指导的PDT的方法称为光学诊疗[张志军,康苗苗,王媛玮,宋光杰,温海飞,王东,唐本忠.聚集诱导发光材料在光学诊疗中的研究进展[J].发光学报,2021,42(03):361-378.],属于诊疗一体化领域的重要分支。然而在传统的PDT治疗过程中往往使用白光或近紫外光光源,意味着其组织穿透能力有限,无法应用于深层肿瘤治疗。由于近红外光(NIR)的组织穿透力非常强,且光源能量较低,对生物组织的伤害较小,在肿瘤成像与治疗领域具有重要的应用前景。
AIEgens的种类有很多,其中线型三苯胺类给体-苯并噻二唑受体-氰基受体(D-A-A)型分子具有合成路线简单和空穴传输性能良好的优点。例如文献(Chen,Yi-Hong,etal."Vacuum-deposited small-molecule organic solar cells with high powerconversion efficiencies by judicious molecular design and deviceoptimization."Journal of the American Chemical Society 134.33(2012):13616-13623.)公开了一种AIEgens分子(结构式如下),该AIEgens分子具有NIR光发射的AIE性能,但是该文未提及是否具有ROS性能,申请人按该文方法合成该分子,结果发现该分子没有NIR光发射的ROS产生性能。
因此,寻找一种具有NIR光发射的AIE性能及ROS产生性能的AIEgens分子具有重要的现实意义。
多肽胶束是由两亲性多肽序列通过自组装形成得到,由于多肽本身具有很好的生物相容性,生物可降解性及无免疫原性等显著优势,将其作为药物载体在生物医学方面具有很广泛的应用前景。此外,通过在分子水平上进行设计和调控多肽自组装行为,使其根据环境刺激(pH值、温度和光照等)的变化而改变自组装体结构与形状,这使其在生物医学领域具有更巨大的应用潜力。目前,尚未有两亲性小分子功能肽和线型三苯胺类给体-苯并噻二唑受体-氰基受体构建的诊疗剂用于肿瘤的特异性诊疗的报道。
发明内容
针对现有技术中存在的不足,本发明以4,4'-二甲氧基-4”-硼酸三苯胺和4-溴-7-(2,2-二氰基乙烯基)苯并[C][1,2,5]噻二唑为主要原料,采用Suzuki Coupling一步反应合成得到AIEgens-1光敏剂;采用多肽固相合成法合成水溶性小分子功能肽(FMOC2-K-GFLGG-R8GD);利用多肽本身的两亲性结构在水溶液中自组装形成胶束,进而实现对疏水性AIEgens-1光敏剂的有效包埋,构建一种酶敏感型多肽胶束型诊疗剂(AIEgens-1@MPA)。本发明的酶敏感型多肽胶束型诊疗剂利用多肽本身的功能多样性,实现AIEgens-1光敏剂在肿瘤细胞中的特异性富集,组织蛋白酶刺激响应性释放及光照刺激诱导的诊疗一体化功能。
为了实现上述目的,本发明采用的技术方案为:
一种酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA,所述诊疗剂AIEgens-1@MPA是球型多肽胶束MPA将疏水性AIEgens-1光敏剂包埋于其疏水腔内;所述球型多肽胶束MPA是由两亲性小分子功能肽FMOC2-K-GFLGG-R8GD在水溶液中自组装形成。
所述AIEgens-1分子,其结构式如下:
分子式:C30H21N5O2S;
分子量:515.59;
所述AIEgens-1分子采取以下合成工艺路线进行合成:
所述AIEgens-1分子的合成方法:在四-(三苯基膦)钯催化下,将摩尔比=1:2的4,4'-二甲氧基-4”-硼酸三苯胺和4-溴-7-(2,2-二氰基乙烯基)苯并[C][1,2,5]噻二唑(4,4'-二甲氧基-4”-硼酸三苯胺:4-溴-7-(2,2-二氰基乙烯基)苯并[C][1,2,5]噻二唑的摩尔比=1:2)进行Suzuki偶联反应,从而有效合成碳-碳键化合物(AIEgens-1)。
所述AIEgens-1分子在水溶液中具有灵敏的AIE性能,并且在AIE光照条件下能够产生大量的ROS,具有PDT治疗效果;
所述AIEgens-1分子在良性溶剂中的荧光强度极弱,这是因为AIEgens-1分子在溶解状态能够自由运动,通过非辐射衰变消耗能量。随着乙腈/水溶液中水体积增加到70%时,AIEgens-1的荧光强度突然显著增强,主要因为AIEgens-1在不良溶剂中的聚集会限制分子内的运动,这时候荧光分子通过辐射跃迁的形式消耗能量,激活了AIE性能。
所述两亲性小分子功能肽FMOC2-K-GFLGG-R8GD,其结构式如下:
分子量:2443.83;
所述两亲性小分子功能肽FMOC2-K-GFLGG-R8GD的多肽序列为:FMOC2-Lys-Gly-Phe-Leu-Gly-Gly-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Gly-Asp。
所述两亲性小分子功能肽FMOC2-K-GFLGG-R8GD是由四种功能性多肽序列构成,分别为能与肿瘤细胞发生特异性粘附的肿瘤靶向肽序列RGD,能快速跨膜进入细胞的细胞穿膜肽序列R8,容易被溶酶体中组织蛋白酶(Cathepsin B)水解断裂的酶敏感性多肽序列GFLGG,及疏水性氨基酸FMOC2-K(氨基酸的2个N端被FMOC基团保护),赋予了胶束较好的自组装性能、主动靶向肿瘤细胞、快速穿膜进入细胞以及酶刺激响应性四种不同功能。
所述两亲性小分子功能肽FMOC2-K-GFLGG-R8GD的制备方法:通过多肽固相Fmoc合成法依次连接天冬氨酸Asp(D),甘氨酸Gly(G),精氨酸Arg(R)×8,甘氨酸Gly(G),甘氨酸Gly(G),亮氨酸Leu(L),苯丙氨酸Phe(F),甘氨酸Gly(G)和赖氨酸Lys(K),用茚三酮的甲醇溶液(10mg/mL)验证树脂上的氨基酸之间酰胺缩合反应是否彻底完成,若验色溶液为亮黄色或者未变色则证明酰胺缩合反应成功。通过脱除氨基保护基团(FMOC)以后,继续下一步酰胺缩合反应,直至合成目标氨基酸序列GFLGG-R8GD,最后接上赖氨酸Lys时,不需要脱去FMOC基团,最终得到目标产物。
所述酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA的制备方法:将AIEgens-1分子、两亲性小分子功能肽FMOC2-K-GFLGG-R8GD溶解于有机溶剂中得溶液,然后将所得溶液加入透析袋,于超纯水中透析除去有机溶剂,两亲性小分子功能肽FMOC2-K-GFLGG-R8GD在水溶液中自组装形成球形胶束MPA,并将疏水性AIEgens-1分子包埋其疏水腔内,透析完成后,将透析袋内的水溶液冷冻干燥成粉末,即得酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA。
进一步,所述有机溶剂为DMF(N,N-二甲基甲酰胺)或DMSO(二甲基亚砜)。
进一步,所述透析袋的截留分子量为1000-3500Da。
进一步,所述透析过程至少为24h。
上述酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA在制备抗肿瘤药物中的应用,所述肿瘤例如是前列腺癌、宫颈癌或乳腺癌等。
由于三苯胺单元具有很强的空穴传输能力,苯并噻二唑和氰基作为吸电子基团能允许更大的电子离域,本发明设计的新型AIEgens-1分子作为一类基于苯并噻二唑的D-A-A型AIEgens分子具有较低的能级带隙(1.97eV),因此具有强的NIR区荧光发射及优异的ROS产生性能。基于在光照条件下AIEgens-1分子能够产生具有细胞毒性的ROS,因此AIEgens-1分子可能实现成像与治疗一体化。
体外释药研究证明了AIEgens-1@MPA诊疗剂在含有组织蛋白酶(Cathepsin B)的环境下具有刺激响应型药物释放行为。本发明对其进行了初步的生物活性评价,结果显示,AIEgens-1@MPA诊疗剂具有明显的特异性进入肿瘤细胞HeLa,并抑制肿瘤细胞生长的作用。
与现有技术相比,本发明的技术方案具有以下优点和有益效果:
1、本发明设计和合成的新型AIEgens-1光敏剂具有合成简单、产率高和NIR(主要在第一近红外NIR-I窗口,650-900nm)荧光发射及优异的ROS产生性能的优点,有利于其在生物医用领域的应用。
2、本发明设计和合成的新型两亲性小分子功能肽(FMOC2-K-GFLGG-R8GD)具有肿瘤细胞靶向性(RGD),细胞穿膜性(R8),组织蛋白酶敏感性(GFLGG)和自组装性能,在水溶液中能够自组装形成核-壳型纳米胶束。该两亲性小分子功能肽的临界胶束浓度值(CMC)为1.9mg/L,远远小于传统的两亲性多肽的CMC值,表明该两亲性小分子功能肽在极低的浓度下能够自组装形成胶束,且纳米胶束的结构更稳定。
3、负载AIEgens-1光敏剂的胶束(AIEgens-1@MPA)的平均粒径为35nm±1nm,具有EPR(高渗透长滞留)效应,在药物载体的制备和药物可控释放等生物领域具有巨大的应用潜力。
4、相比常用的聚合物胶束,多肽胶束具有无毒,生物相容性好、生物可降解性及功能多样化等优势。材料的结构组成简单,能有效避免组成结构复杂的治疗系统中效率低、系统副作用大的不良后果,为精准的可视化肿瘤诊疗提供新的研究思路。
附图说明
图1为实施例一合成的AIEgens-1的1HNMR图谱;
图2为实施例一合成的AIEgens-1的MALDI-TOF MS图谱;
图3为实施例二合成的FMOC2-K-GFLGG-R8GD的MALDI-TOF MS图谱;
图4为实施例二合成的FMOC2-K-GFLGG-R8GD的CMC图;
图5为实施例三合成的AIEgens-1@MPA的TEM图谱;
图6为实施例四中AIEgens-1的AIE性能测试实验结果图;
图7为实施例五中AIEgens-1@MPA的ROS产生性能测试实验结果图;
图8为实施例六中AIEgens-1@MPA的体外细胞内吞实验结果图;
图9为实施例六中AIEgens-1@MPA的肿瘤细胞内产生ROS性能实验结果图;
图10为实施例六中AIEgens-1@MPA的细胞暗毒性实验结果图,其中,*:p<0.05,**:p<0.01;
图11为实施例六中AIEgens-1@MPA的细胞光毒性实验结果图,其中,*:p<0.05,**:p<0.01。
具体实施方式
下面申请人将结合具体的实施例和附图对本发明的技术方案做详细说明,便于本领域技术人员清楚地理解本发明。但是以下实施例不应以任何方式被解释为对本发明权利要求书请求保护的范围的限制。
以下实施例中所用试剂均为常见市售商品,纯度级别均为分析纯。
实施例一、AIEgens-1的合成
向三口烧瓶中加入1mol 4-溴-7-(2,2-二氰基乙烯基)苯并[C][1,2,5]噻二唑,2mol 4,4'-二甲氧基-4”-硼酸三苯胺,10mmol无水碳酸钠(1.06g),0.04mmol四-(三苯基膦)钯(50mg),通入氮气以除尽烧瓶中的氧气。在氮气氛围下向三口烧瓶中加入20mL甲苯和3mL去离子水,于110℃条件下回流反应4h。待反应结束后,加入40mL二氯甲烷和4.0g硅胶粉,将得到的产物旋转浓缩成粉末。随后将其置于60℃真空干燥箱中,烘干得到粗产物。采用硅胶柱层析法进行产物分离,用体积比为4:1的二氯甲烷/石油醚混合溶剂作为洗脱剂,最终纯化并烘干得到紫黑色固体(收率:51%)。其结构通过核磁氢谱验证(见图1)。1H NMR(400MHz,CDCl3)δ3.83(s,6H),6.90(d,J=8.0Hz,4H),7.03(d,J=8.0Hz,2H),7.15(d,J=8.0Hz,4H),7.83(d,J=8.0Hz,H),7.95(d,J=8.0Hz,2H),8.78(d,J=8.0Hz,1H),8.83(s,1H)。利用基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)检测其分子量,实际测得分子量为551.03,见图2,与理论值符合(理论分子量为551.59)。结合MALDI-TOF MS图谱和1HNMR数据,证明成功合成了AIEgens-1光敏剂。
实施例二、两亲性小分子肽(FMOC2-K-GFLGG-R8GD)的合成
采用标准的Fmoc固相多肽合成法合成FMOC2-K-GFLGG-R8GD:称取2-氯-三苯甲基氯树脂(0.8g,0.97mmol/g)加入多肽固相合成柱中,加入20mLN,N-二甲基甲酰胺(DMF)浸泡树脂30分钟,使其充分溶胀后抽滤除去DMF。准备接入第一个氨基酸,进入氨基酸缩合反应。随后向多肽固相合成柱中加入溶有Fmoc-Asp(OtBu)-OH(相当于树脂取代度的3倍量)和DIEA(相当于树脂取代度的6当量)的DMF溶液,反应2h。反应结束后抽走滤液,并用DMF洗涤4次。得到第一个被FMOC保护的氨基酸缩合后的肽树脂。然后将20%的哌啶(piperidine)/DMF(v/v)溶液加到树脂中,脱除氨基酸N端的氨基保护基FMOC基团,反应半小时(15min×2)后除去反应溶液,并用DMF洗涤树脂三次。此后每一次在树脂上连接氨基酸残基时,均将溶解有被FMOC保护的氨基酸(相当于树脂取代度的4倍量)、1-羟基苯并三唑(HOBt,相当于树脂取代度的4倍量)、苯并三氮唑-N,N,N’N’-四甲基脲六氟磷酸盐(HBTU,相当于树脂取代度的4倍量)和N,N-二异丙基乙胺(DIEA,相当于树脂取代度的6倍量)的DMF加入到树脂中进行缩合反应。反应1.5h后,用茚三酮的甲醇溶液(10mg/mL)验证树脂上的氨基酸之间酰胺缩合反应是否彻底完成,若验色溶液为亮黄色或者未变色则证明酰胺缩合反应成功。酰胺缩合反应成功后,脱除FMOC基团以后,继续进行酰胺缩合反应,依次与FMOC-Gly-OH,FMOC-Arg(Pbf)-OH(与FMOC-Arg(Pbf)-OH重复八次),FMOC-Gly-OH,FMOC-Gly-OH,FMOC-Leu-OH,FMOC-Phe-OH,FMOC-Gly-OH,FMOC-Lys(Boc)-OH)进行酰胺缩合反应,直至合成目标氨基酸序列。待多肽序列合成完毕,依次用DMF和DCM洗涤树脂3次,并将树脂真空干燥过夜。用配好的多肽切落剂(三氟乙酸:水:三异丙基硅烷=95:2.5:2.5,v/v/v)加入到树脂中,在室温下搅拌反应2h。收集滤液并进行浓缩得到浓缩液,然后选用冰乙醚使多肽粗产品从中沉淀出来,最后将沉淀置于真空干燥箱中进行干燥,得到白色粉末产物。通过基质辅助激光解析-飞行时间质谱(MALDI-TOF MS)检测所得白色粉末的分子量(见图3)。FMOC2-K-GFLGG-R8GD小分子肽的理论分子量为2443.83,实际测得分子量为2443.72,证明FMOC2-K-GFLGG-R8GD多肽的成功合成。在MALDI-TOF谱图中,分子量为2540.81处的峰为FMOC2-K-GFLGG-R8GD与一个三氟乙酸分子之间发生缩合反应,脱去一个H2O分子得到的产物。
以芘为疏水性荧光探针,利用荧光分光光度计测FMOC2-K-GFLGG-R8GD的临界胶束浓度(CMC)值。精确称量9.7mg疏水性芘荧光分子,将其溶于10mL丙酮,然后稀释至4.6×10- 6mol/L,备用。往3.6mL已配好的一系列浓度的FMOC2-K-GFLGG-R8GD多肽水溶液(取FMOC2-K-GFLGG-R8GD多肽粉末溶于去离子水中配置而成)加入45μL4.6×10-6mol/L芘的丙酮溶液,配置成样品。将配置好的样品放入37℃摇床(150r/min)震荡24h,确保丙酮完全挥发。最后,利用荧光分光光度计测定样品溶液在360-400nm内的发射光谱(激发波长为342nm,发射光和激发光的狭缝宽度均设为5nm)。以FMOC2-K-GFLGG-R8GD溶液在390nm与360nm处的荧光强度比值(I390/I360)作为纵坐标,以FMOC2-K-GFLGG-R8GD溶液浓度的对数(lgC)为横坐标作图。找出两条直线的交点对应的横坐标值,从而计算得到该胶束的临界胶束浓度(CMC)。FMOC2-K-GFLGG-R8GD的CMC值测得为1.9mg/L(见图4),远远小于传统的两亲性多肽的CMC值。表明该两亲性小分子功能肽在极低的浓度下能自组装形成胶束,且结构稳定,非常适合于药物载体的设计和制备。
实施例三、AIEgens-1@MPA的合成
将50mg AIEgens-1(实施例一制备的)、50mg FMOC2-K-GFLGG-R8GD(实施例二制备的)和20mL DMF加入单口圆底烧瓶中,室温下搅拌反应24h。随后将上述溶液加入透析袋(MWCO 1000Da),于超纯水中透析24h,每隔4h换一次超纯水。透析完后,将透析袋内的水溶液冻成固体,然后置于冻干机冷冻干燥成粉末状。利用透射电子显微镜(TEM)表征FMOC2-K-GFLGG-R8GD小分子肽自组装形成胶束的形貌和尺寸,如图5所示。从图5TEM中观察到胶束呈球形,并具有较好的分散性,胶束的干态粒径为35nm±1nm,证明了两亲性小分子肽FMOC2-K-GFLGG-R8GD在水溶液中具有较好的自组装性能。
实施例四、AIE性能测试实验
配制一系列AIEgens-1(实施例一制备的)浓度为30mg/L的AIEgens-1溶液(以乙腈-水作溶剂),其中水与乙腈的体积比分别为95%、90%、80%、70%、60%、50%、40%、30%、20%、10%和0%。设定激发波长为572nm,利用荧光分光光度计测定以上溶液在600-650nm范围内的荧光发射光谱(见图6A),并记录其在606nm处的荧光强度值(见图6B)。AIEgens-1在良性溶剂(乙腈)中,在606nm处的荧光强度极弱。随着乙腈/水混合溶液中水的体积比(Water Fraction,指水与乙腈的体积比)增加到70%时,AIEgens-1在606nm处的荧光强度突然显著增强,激活了AIE过程。当水的体积比为90%和95%时,AIEgens-1的荧光强度分别是在纯乙腈中的荧光强度的36倍和54倍,证明了我们合成的荧光小分子AIEgens-1具有良好的AIE效应。
实施例五、ROS产生性能测试实验
首先,取0.5mLNaOH(10mmol/L)溶液加至100uL DCFH-DA溶液(10mmol/L)中,在室温条件下避光静置0.5h;再将以上溶液加至0.5mLpH为7.4的PBS(0.2mol/L)溶液中,配制得到DCFH。取实施例三合成的冻干粉(AIEgens-1@MPA材料)溶于去离子水中,配制50mg/LAIEgens-1@MPA溶液。随后,将DCFH加入50mg/L AIEgens-1@MPA溶液中,使DCFH的最终浓度为20μmol/L。将以上溶液分成两份,将第一份溶液置于红外光(680nm,30mW cm-2)下照射,并在预定的时间间隔用荧光分光光度计测定AIEgens-1@MPA在525nm处的荧光强度值。第二份溶液则在避光条件下重复上述荧光检测的方法。如图7所示,第一份溶液经过红外光照120s以后,AIEgens-1@MPA在525nm处的荧光强度比初始的荧光强度高出17倍。而避光条件下的AIEgens-1@MPA在525nm处的荧光强度在此期间基本没有变化。证明了AIEgens-1@MPA在特定波长的光照刺激下,能够在短时间内产生大量的ROS,所以具备杀死细胞的潜力。但是,在无光照刺激的情况下,AIEgens-1@MPA产生的ROS很少。因此,我们可以通过调整光照时间的长短,实现有效调控AIEgens-1@MPA产生PDT治疗的效果。
实施例六、体外细胞实验
人宫颈癌细胞(HeLa)和非洲绿猴肾细胞(COS 7)均为本申请发明人实验室的冻存材料,细胞培养基则由1%的双抗(青霉素-链霉素)加入购买的含10%的胎牛血清蛋白的DMEM培养基中配制而成(以下简称DMEM)。所有细胞均在细胞培养基中孵育,在37℃条件下,含有5%CO2的细胞培养箱中培养。
体外细胞内吞实验:将HeLa细胞和COS 7细胞分别接种到共聚焦培养皿中培养24h,然后各加入1mL含有AIEgens-1@MPA诊疗剂(50mg/L)的DMEM溶液,并将共聚焦培养皿放入细胞培养箱中继续培养4h。随后用pH为7.4的PBS溶液(10mmol/L)洗涤细胞三次,再加入1mL含Hoechst 33342(10μg/mL)的DMEM(不含胎牛血清)对细胞核进行染色15min。完成细胞核染色后吸出培养基,用pH为7.4的PBS溶液(10mmol/L)洗涤细胞三次,加入2mL DMEM培养基,并通过超分辨显微镜观测(激发波长:552nm,荧光接收通道:680nm±30nm)。
图8为实施例六中AIEgens-1@MPA的体外细胞内吞实验结果图;图8从左至右依次为明场(Bright),细胞核的Hoechst 33342蓝色荧光场(Hoechst 33342),AIEgens-1@MPA的红色荧光场(AIEgens-1@MPA)和叠加场(Merge)。从上至下依次为与AIEgens-1@MPA诊疗剂共培养的COS7细胞、与AIEgens-1@MPA诊疗剂共培养的HeLa细胞。从图8所知,HeLa细胞中出现了大量AIEgens-1的红色荧光,这是由于AIEgens-1@MPA诊疗剂表面修饰的靶向肽RGD,能够识别并与HeLa细胞表面过度表达的整合素αvβ3结合,并且在穿膜肽R8的辅助下快速穿膜进入HeLa细胞,从而将AIEgens-1光敏剂有效运输到细胞质内。接着,HeLa细胞质内过表达的CathepsinB能有效裂解GFLGG序列,从而引发多肽胶束结构瓦解,实现AIEgens-1的定点释放。与此同时,疏水性AIEgens-1光敏剂在细胞内会迅速团聚,激活其AIE效应,进而发挥细胞内成像功能。相比而言,AIEgens-1@MPA在正常细胞(COS7)中的荧光强度很弱,有效证明了RGD肽的靶向性能。
肿瘤细胞内产生ROS性能实验:将HeLa细胞接种于共聚焦培养皿中培养24h,随后加入1mL含有AIEgens-1@MPA诊疗剂(50mg/L)的DMEM溶液,并继续共培养4h后。然后加入1mL配制好的DCFH-DA溶液(20μmol/L),继续共培养30min。以不加AIEgens-1@MPA的DMEM溶液与HeLa细胞共培养为对照组。最后,将共聚焦培养皿置于近红外光照(680nm,150mW cm-2)下照射1min,用pH为7.4的PBS(10mmol/L)溶液洗涤细胞三次,并通过超分辨显微镜观测(激发波长:488nm,荧光接收通道:525nm±10nm)。
图9为实施例六中AIEgens-1@MPA的肿瘤细胞内产生ROS性能实验结果图;图9从左至右依次为明场(Bright),DCF的绿色荧光场(DCF)和叠加场(Merge)。从上至下依次为没经过任何处理的HeLa细胞(Blank),在避光条件下与AIEgens-1@MPA诊疗剂共培养的HeLa细胞(AIEgens-1@MPA),和在光照条件下与AIEgens-1@MPA诊疗剂共培养的HeLa细胞(AIEgens-1@MPA+Light)。如图9所示,近红外光照下的HeLa细胞中呈现强烈的DCF绿色荧光,这是由于AIEgens-1@MPA胶束将AIEgens-1光敏剂靶向运输至HeLa细胞以后,680nm光照会诱导其在细胞内产生大量的ROS,从而将不发光的DCFH氧化为发绿色荧光的DCF。相比而言,在避光条件下的HeLa细胞中基本上没有出现DCF绿色荧光,进一步证明了NIR光照能诱导AIEgens-1@MPA胶束在细胞内有效产生ROS,具有PDT治疗的生物医用潜能。
细胞毒性实验:采用CCK-8法评估AIEgens-1@MPA诊疗剂分别对正常细胞和肿瘤细胞的毒性大小。
AIEgens-1@MPA的细胞光毒性实验:将HeLa细胞以5000个/孔的密度接种在96孔板中并加入100μL含有10%FBS的DMEM培养基,于37℃下在5%CO2培养箱中培养48h。然后,每孔加入100μL含有固定浓度的AIEgens-1@MPA诊疗剂的DMEM培养基。培养4h后,将其置于近红外光照(680nm,150mW cm-2)下照射3min,再放回培养箱中继续培养24h,用酶标仪检测每个孔在460nm处的吸光度值(OD)。随后吸出所有培养基,再加入100μL新鲜DMEM培养基和20μLCCK-8试剂于96孔板各个孔中,于37℃下在细胞培养箱中培养4h,使用酶标仪第二次检测每个板在460nm处的OD,并计算出细胞的存活率。以上操作未作说明的均在避光条件下进行。
AIEgens-1@MPA的细胞暗毒性实验:暗毒性实验同AIEgens-1@MPA的细胞光毒性实验一样,唯一不同在于“将培养基置于近红外光照(680nm,150mW cm-2)下照射3min”这一操作调整为“继续在培养箱中培养”,即暗毒性实验全程均在避光条件下进行。
细胞光毒性实验、细胞暗毒性实验中检测得到的OD值均基于4个独立平行样的平均值,结果表示为平均值±标准偏差(SD)。细胞的相对存活率根据以下公式可计算得到:
Viability(%)=(ODtreated/ODcontrol)×100
其中,ODcontrol为CCK-8试剂材料加入96孔板前检测到的吸光度值,ODtreated为CCK-8试剂材料加入96孔板后检测到的吸光度值。
图10为实施例六中AIEgens-1@MPA的细胞暗毒性实验结果图,其中,*:p<0.05,**:p<0.01。如图所示,在避光条件下,AIEgens-1@MPA诊疗剂含量在0-50mg/L(μg/mL)的范围内对正常细胞(COS 7)和肿瘤细胞(HeLa)的均没有毒性,AIEgens-1@MPA诊疗剂含量在0-25mg/L的范围内细胞存活率都在90%以上,表明AIEgens-1@MPA诊疗剂在无光照刺激条件下具有很好的生物相容性。
但是,在NIR光照下(图11为实施例六中AIEgens-1@MPA的细胞光毒性实验结果图,其中,*:p<0.05,**:p<0.01),AIEgens-1@MPA诊疗剂在浓度为25mg/L时与HeLa细胞共培养以后的细胞存活率下降至68%左右,当浓度为50mg/L时,HeLa细胞的存活率甚至下降至20%左右(见图11)。与此同时,COS 7细胞的存活率仍在90%左右。
以上细胞毒性实验足以证明AIEgens-1@MPA诊疗剂表面修饰的RGD肽对肿瘤细胞的靶向性和近红外光照引发的PDT治疗性能。
Claims (8)
2.根据权利要求1所述的酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA,其特征在于,所述酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA的制备方法包括以下步骤:将AIEgens-1、两亲性小分子功能肽FMOC2-K-GFLGG-R8GD溶解于有机溶剂中得溶液,然后将所得溶液加入透析袋,于超纯水中透析除去有机溶剂,两亲性小分子功能肽FMOC2-K-GFLGG-R8GD在水溶液中自组装形成球形胶束MPA,将疏水性AIEgens-1分子包埋于其疏水腔内,透析完成后,将透析袋内的水溶液冷冻干燥成粉末,得到酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA。
3.根据权利要求2所述的酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA,其特征在于,所述透析袋的截留分子量为1000-3500Da。
4.根据权利要求2所述的酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA,其特征在于,所述有机溶剂为DMF或DMSO。
6.根据权利要求5所述的酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA,其特征在于,所述AIEgens-1的合成方法包括以下步骤:在四-(三苯基膦)钯催化下,将摩尔比=1:2的4,4'-二甲氧基-4″-硼酸三苯胺和4-溴-7-(2,2-二氰基乙烯基)苯并[C][1,2,5]噻二唑进行Suzuki偶联反应得到。
7.权利要求1-6任一所述的酶敏感型多肽胶束型诊疗剂AIEgens-1@MPA在制备抗肿瘤药物中的应用。
8.根据权利要求7所述的应用,其特征在于:所述肿瘤为前列腺癌、宫颈癌或乳腺癌。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110540998.4A CN113209020B (zh) | 2021-05-18 | 2021-05-18 | 一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110540998.4A CN113209020B (zh) | 2021-05-18 | 2021-05-18 | 一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113209020A CN113209020A (zh) | 2021-08-06 |
CN113209020B true CN113209020B (zh) | 2022-05-03 |
Family
ID=77092786
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110540998.4A Active CN113209020B (zh) | 2021-05-18 | 2021-05-18 | 一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113209020B (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113651872A (zh) * | 2021-08-09 | 2021-11-16 | 大连理工大学 | 一类基于两亲性多肽的自组装纳米光敏剂及其制备方法与应用 |
CN114404364B (zh) * | 2022-01-18 | 2023-06-16 | 广西大学 | 一种可诱导肿瘤相关巨噬细胞m1型极化的光敏纳米胶束的构建及其抗肿瘤应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108864106A (zh) * | 2018-07-17 | 2018-11-23 | 南京邮电大学 | 新型近红外二区有机小分子荧光探针的制备与应用 |
CN108948152A (zh) * | 2017-05-18 | 2018-12-07 | 中国科学院上海药物研究所 | 一种两亲性穿膜肽键合物、其制备方法及用途 |
CN111362971A (zh) * | 2020-03-16 | 2020-07-03 | 北京师范大学 | 靶向psma的双苯并噻二唑类化合物及其制备方法与应用 |
CN111471043A (zh) * | 2020-04-21 | 2020-07-31 | 浙江工业大学 | 一种含有苯并[c][1,2,5]噻二唑衍生物受体结构单元的有机发光材料及应用 |
-
2021
- 2021-05-18 CN CN202110540998.4A patent/CN113209020B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108948152A (zh) * | 2017-05-18 | 2018-12-07 | 中国科学院上海药物研究所 | 一种两亲性穿膜肽键合物、其制备方法及用途 |
CN108864106A (zh) * | 2018-07-17 | 2018-11-23 | 南京邮电大学 | 新型近红外二区有机小分子荧光探针的制备与应用 |
CN111362971A (zh) * | 2020-03-16 | 2020-07-03 | 北京师范大学 | 靶向psma的双苯并噻二唑类化合物及其制备方法与应用 |
CN111471043A (zh) * | 2020-04-21 | 2020-07-31 | 浙江工业大学 | 一种含有苯并[c][1,2,5]噻二唑衍生物受体结构单元的有机发光材料及应用 |
Non-Patent Citations (1)
Title |
---|
Self-assembled micelles of a multi-functional amphiphilic fusion (MFAF) peptide for targeted cancer therapy;Yin-Jia Cheng等;《Polymer Chemistry》;20151231;全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN113209020A (zh) | 2021-08-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113209020B (zh) | 一种酶敏感型多肽胶束型诊疗剂及其在制备抗肿瘤药物中的应用 | |
CN109096170B (zh) | 近红外染料、其靶向成像剂、纳米载体和抗癌药物及应用 | |
Lin et al. | A pH-responsive stellate mesoporous silica based nanophotosensitizer for in vivo cancer diagnosis and targeted photodynamic therapy | |
CN107118765B (zh) | 一种核仁靶向荧光碳点及其制备方法与应用 | |
CN104195176A (zh) | 一种超分子杂化肽类树状大分子自组装体及其制备方法与应用 | |
CN110179754B (zh) | 具有氧化还原响应性且可增强组织渗透的多功能脂质体 | |
Yang et al. | Magainin II modified polydiacetylene micelles for cancer therapy | |
CN114044898B (zh) | 一种赖氨酸接枝聚乙烯亚胺阳离子基因载体及其制备方法和应用 | |
Wang et al. | Real-time self-tracking of an anticancer small molecule nanodrug based on colorful fluorescence variations | |
Xue et al. | Synthesis and biological evaluation of an epidermal growth factor receptor-targeted peptide-conjugated phthalocyanine-based photosensitiser | |
CN113429461B (zh) | 一种聚集诱导发光多肽胶束型诊断试剂及其在近红外区域生物成像中的应用 | |
Accardo et al. | Amphiphilic CCK peptides assembled in supramolecular aggregates: structural investigations and in vitro studies | |
CN113144171B (zh) | 一种氧化响应形貌转变多肽纳米药物 | |
CN111548419B (zh) | 靶向ddr2的多肽及其应用 | |
Ding et al. | Novel oligopeptide nanoprobe for targeted cancer cell imaging | |
CN115385861B (zh) | 一种荧光探针及其制备方法和应用 | |
CN112375562A (zh) | 一类基于半胱氨酸-多胺-吗啉修饰的量子点溶酶体靶向荧光探针及其制备方法和应用 | |
Chen et al. | Ligand‐functionalization of BPEI‐coated YVO4: Bi3+, Eu3+ nanophosphors for tumor‐cell‐targeted imaging applications | |
US10927221B2 (en) | Dendrimeric metallacrowns | |
CN117017938A (zh) | 一种用于铁死亡治疗的多功能纳米平台及其制备方法和应用 | |
CN115010786B (zh) | 一种gsh-乏氧微环境双响应的自组装荧光探针及其制备方法和应用 | |
CN115814111A (zh) | 一种近红外荧光adc免疫制剂及其制备方法与应用 | |
CN109701013B (zh) | 一种靶向纳米给药系统及制备方法 | |
Kougioumtzi et al. | Development of novel GnRH and Tat 48–60 based luminescent probes with enhanced cellular uptake and bioimaging profile | |
CN111423497B (zh) | 拮抗肽、其共聚物及纳米组装体、及其制备方法和应用 |
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 |