CN101785865A - pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof - Google Patents
pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof Download PDFInfo
- Publication number
- CN101785865A CN101785865A CN201010118032A CN201010118032A CN101785865A CN 101785865 A CN101785865 A CN 101785865A CN 201010118032 A CN201010118032 A CN 201010118032A CN 201010118032 A CN201010118032 A CN 201010118032A CN 101785865 A CN101785865 A CN 101785865A
- Authority
- CN
- China
- Prior art keywords
- response type
- micelle
- grafting
- polyphosphazene
- polyphosphazenes
- 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
- 229920002627 poly(phosphazenes) Polymers 0.000 title claims abstract description 97
- 230000004044 response Effects 0.000 title claims abstract description 85
- 239000000693 micelle Substances 0.000 title claims abstract description 80
- 238000002360 preparation method Methods 0.000 title claims description 48
- 239000003814 drug Substances 0.000 claims abstract description 69
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 20
- 239000000843 powder Substances 0.000 claims abstract description 9
- 229920000642 polymer Polymers 0.000 claims description 61
- 239000000243 solution Substances 0.000 claims description 42
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 37
- 239000000203 mixture Substances 0.000 claims description 34
- NWXMGUDVXFXRIG-WESIUVDSSA-N (4s,4as,5as,6s,12ar)-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O NWXMGUDVXFXRIG-WESIUVDSSA-N 0.000 claims description 33
- 229930195573 Amycin Natural products 0.000 claims description 33
- 229940079593 drug Drugs 0.000 claims description 27
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 9
- 238000004108 freeze drying Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000002246 antineoplastic agent Substances 0.000 claims description 8
- 239000003960 organic solvent Substances 0.000 claims description 8
- 238000009826 distribution Methods 0.000 claims description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 229940041181 antineoplastic drug Drugs 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 5
- 239000000337 buffer salt Substances 0.000 claims description 4
- 229930012538 Paclitaxel Natural products 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 229960001592 paclitaxel Drugs 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims description 3
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 claims description 2
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims description 2
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 claims description 2
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 claims description 2
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin 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(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 claims description 2
- 229960000975 daunorubicin Drugs 0.000 claims description 2
- 230000006837 decompression Effects 0.000 claims description 2
- 229960003668 docetaxel Drugs 0.000 claims description 2
- 229960000485 methotrexate Drugs 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 abstract description 16
- 210000004881 tumor cell Anatomy 0.000 abstract description 11
- 238000000034 method Methods 0.000 abstract description 10
- 239000006185 dispersion Substances 0.000 abstract description 6
- 230000008685 targeting Effects 0.000 abstract description 3
- 238000000502 dialysis Methods 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000000259 anti-tumor effect Effects 0.000 abstract 2
- 230000000973 chemotherapeutic effect Effects 0.000 abstract 1
- 230000036571 hydration Effects 0.000 abstract 1
- 238000006703 hydration reaction Methods 0.000 abstract 1
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 54
- 210000004027 cell Anatomy 0.000 description 28
- 229920001223 polyethylene glycol Polymers 0.000 description 20
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 20
- 238000006243 chemical reaction Methods 0.000 description 18
- 231100000135 cytotoxicity Toxicity 0.000 description 16
- 230000003013 cytotoxicity Effects 0.000 description 16
- 230000000694 effects Effects 0.000 description 13
- 239000002202 Polyethylene glycol Substances 0.000 description 12
- 229920001577 copolymer Polymers 0.000 description 11
- 239000002585 base Substances 0.000 description 10
- -1 MRP) Proteins 0.000 description 9
- 230000036457 multidrug resistance Effects 0.000 description 9
- 230000003578 releasing effect Effects 0.000 description 8
- 125000001424 substituent group Chemical group 0.000 description 8
- 150000001875 compounds Chemical class 0.000 description 7
- FKRCODPIKNYEAC-UHFFFAOYSA-N ethyl propionate Chemical compound CCOC(=O)CC FKRCODPIKNYEAC-UHFFFAOYSA-N 0.000 description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 206010059866 Drug resistance Diseases 0.000 description 6
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 6
- 238000010189 synthetic method Methods 0.000 description 6
- 229960005274 benzocaine Drugs 0.000 description 5
- MTZQAGJQAFMTAQ-UHFFFAOYSA-N benzoic acid ethyl ester Natural products CCOC(=O)C1=CC=CC=C1 MTZQAGJQAFMTAQ-UHFFFAOYSA-N 0.000 description 5
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 5
- 238000006116 polymerization reaction Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- JWYUFVNJZUSCSM-UHFFFAOYSA-N 2-aminobenzimidazole Chemical compound C1=CC=C2NC(N)=NC2=C1 JWYUFVNJZUSCSM-UHFFFAOYSA-N 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Natural products CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- 239000010408 film Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 125000001165 hydrophobic group Chemical group 0.000 description 4
- 230000031700 light absorption Effects 0.000 description 4
- 239000008176 lyophilized powder Substances 0.000 description 4
- 150000003141 primary amines Chemical class 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- PJWZTMATHPXZCW-UHFFFAOYSA-N 1-butyl-n-methylpiperidin-4-amine Chemical class CCCCN1CCC(NC)CC1 PJWZTMATHPXZCW-UHFFFAOYSA-N 0.000 description 3
- KDHWOCLBMVSZPG-UHFFFAOYSA-N 3-imidazol-1-ylpropan-1-amine Chemical class NCCCN1C=CN=C1 KDHWOCLBMVSZPG-UHFFFAOYSA-N 0.000 description 3
- 102100033350 ATP-dependent translocase ABCB1 Human genes 0.000 description 3
- 206010006187 Breast cancer Diseases 0.000 description 3
- 208000026310 Breast neoplasm Diseases 0.000 description 3
- 102000003923 Protein Kinase C Human genes 0.000 description 3
- 108090000315 Protein Kinase C Proteins 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 3
- 229920001400 block copolymer Polymers 0.000 description 3
- 201000008275 breast carcinoma Diseases 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000001647 drug administration Methods 0.000 description 3
- 238000012377 drug delivery Methods 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 229920000578 graft copolymer Polymers 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Substances C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000000954 titration curve Methods 0.000 description 3
- 238000001291 vacuum drying Methods 0.000 description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 108010047230 Member 1 Subfamily B ATP Binding Cassette Transporter Proteins 0.000 description 2
- 229940034982 antineoplastic agent Drugs 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 229920005601 base polymer Polymers 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000012531 culture fluid Substances 0.000 description 2
- 210000001163 endosome Anatomy 0.000 description 2
- GKCXXDSWWDWUHS-BYPYZUCNSA-N ethyl (2s)-2-amino-3-hydroxypropanoate Chemical compound CCOC(=O)[C@@H](N)CO GKCXXDSWWDWUHS-BYPYZUCNSA-N 0.000 description 2
- NTNZTEQNFHNYBC-UHFFFAOYSA-N ethyl 2-aminoacetate Chemical compound CCOC(=O)CN NTNZTEQNFHNYBC-UHFFFAOYSA-N 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000003969 glutathione Nutrition 0.000 description 2
- 229920001477 hydrophilic polymer Polymers 0.000 description 2
- 210000003712 lysosome Anatomy 0.000 description 2
- 230000001868 lysosomic effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 239000013638 trimer Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 102100022595 Broad substrate specificity ATP-binding cassette transporter ABCG2 Human genes 0.000 description 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 102000005720 Glutathione transferase Human genes 0.000 description 1
- 108010070675 Glutathione transferase Proteins 0.000 description 1
- 101000823298 Homo sapiens Broad substrate specificity ATP-binding cassette transporter ABCG2 Proteins 0.000 description 1
- 102000004855 Multi drug resistance-associated proteins Human genes 0.000 description 1
- 108090001099 Multi drug resistance-associated proteins Proteins 0.000 description 1
- YPIGGYHFMKJNKV-UHFFFAOYSA-N N-ethylglycine Chemical compound CC[NH2+]CC([O-])=O YPIGGYHFMKJNKV-UHFFFAOYSA-N 0.000 description 1
- 229910003251 Na K Inorganic materials 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 241001337451 Polyblastus cancer Species 0.000 description 1
- 101710183280 Topoisomerase Proteins 0.000 description 1
- 102000007537 Type II DNA Topoisomerases Human genes 0.000 description 1
- 108010046308 Type II DNA Topoisomerases Proteins 0.000 description 1
- 238000004847 absorption spectroscopy Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- BLFLLBZGZJTVJG-UHFFFAOYSA-N benzocaine Chemical compound CCOC(=O)C1=CC=C(N)C=C1 BLFLLBZGZJTVJG-UHFFFAOYSA-N 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 229960001760 dimethyl sulfoxide Drugs 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000000806 elastomer Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- ACHWEOLUTJAHLV-ZETCQYMHSA-N ethyl (2s)-2-amino-3-(1h-imidazol-5-yl)propanoate Chemical compound CCOC(=O)[C@@H](N)CC1=CN=CN1 ACHWEOLUTJAHLV-ZETCQYMHSA-N 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- JUOXYWXXPHUSAI-UHFFFAOYSA-N ethylamino acetate Chemical compound CCNOC(C)=O JUOXYWXXPHUSAI-UHFFFAOYSA-N 0.000 description 1
- 125000000031 ethylamino group Chemical group [H]C([H])([H])C([H])([H])N([H])[*] 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229960004194 lidocaine Drugs 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000010550 living polymerization reaction Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 238000010534 nucleophilic substitution reaction Methods 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 125000004437 phosphorous atom Chemical group 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229920001513 poly[2-(diethylamino)ethyl methacrylate] polymer Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 238000000710 polymer precipitation Methods 0.000 description 1
- 238000003918 potentiometric titration Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 229940121649 protein inhibitor Drugs 0.000 description 1
- 239000012268 protein inhibitor Substances 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000012779 reinforcing material Substances 0.000 description 1
- 238000007151 ring opening polymerisation reaction Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 150000003512 tertiary amines Chemical group 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a pH response type amphipathic stem-grafting polyphosphazenes feeding micelle containing 70%-99.999% of pH response type amphipathic stem-grafting polyphosphazenes and 0.001%-30% of at least one hydrophobic medicament by the total weight of the medicament and the pH response type amphipathic stem-grafting polyphosphazenes, wherein the hydrophobic medicament is a hydrophobic anti-tumor medicament or a hydrophobic chemotherapeutic medicament. The micelle can be prepared by using a film hydration method or a dialysis method and can exist in a state of aqueous dispersion or frozen dry powder. The medicament of the micelle has pH response when released, the micelle can rapidly release the medicament nearby tumor tissues or in tumor cells at fixed points to reverse the multi-medicament resistance of the tumor cells, and meanwhile, the amphipathic stem-grafting polyphosphazenes has good biocompatibility and less toxicity and is suitable for being used as a targeting carrier of the anti-tumor medicament.
Description
Technical field
The present invention relates to drug delivery system, be specifically related to a kind of pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof.
Background technology
(multidrug resistance MDR) is meant tumor cell to a kind of chemical sproof while of antineoplastic agent deposits yields to tumor multidrug-resistance, to the phenomenon of structure and diverse other antineoplastic agent deposits yields cross resistances of mechanism of action.The formation mechanism of MDR is very complicated, both with cell membrane on multidrug-associated protein (multidrug resistance associated protein, MRP), P glycoprotein (P-glycoprotein, P-gp), lung multidrug-associated protein (LRP), breast carcinoma drug resistance associated protein memebrane proteins such as (BCRP) is relevant, simultaneously also with intracytoplasmic DNA topoisomerase II (topoisomerase, Topo II), Protein kinase C (proteinkinase C, PKC), glutathion (glutathione, GSH) and glutathion-s-transferring enzyme (glutathione-s-transferase, relevant (the Ambudkar SV of enzyme such as GST), Dey S, HrycynaCA, Ramachandra M, Pastan I, Gottesman MM.Annu.Rev.Pharmacol.Toxicol.1999,39,361～398; Hunter J, Hirst BH.Adv.Drug Deliv.Rev.1997,25,129～157; Meschini S, Marra M, Calcabrini A.Toxicol.In Vitro 2002,16,389～398).The generation of MDR is the one of the main reasons that causes the chemotherapy of tumors failure, particularly newfound in recent years tumor stem cell (the tumor stem cells that in tumor differentiation and breeding, plays central role, TSC) generally have multidrug resistance especially, so the multidrug resistance of reversing tumor cell just becomes the important topic that current cancer research and treatment field need to be resolved hurrily.
Although people have also found the number of chemical medicine and can suppress the activity of the memebrane protein relevant with multidrug resistance, yet thereby since these medicines generally have very strong toxic and side effects and limited its clinical practice; On the other hand, utilize the theory and practice of gene therapy reversal of multidrug resistance of tumor cells all also ripe far away.Therefore, thus be exactly that the drug level that increases tumor cell treatment target spot place is as much as possible offset MDR effluxing and Detoxication medicine at present at the most feasible countermeasure of MDR.Thereby ideal, as can effectively to resist tumor multidrug-resistance medicine-carried system should be directed be enriched in the tumor tissues place and promptly discharge entrained medicine immediately overcomes MDR.Administration nano-drug administration system such as liposome and micelle can by passive, initiatively or physicochemical mechanism of action realize the cancer target administration, yet common administration nano-drug administration system does not have specificity on drug release behavior, though pharmaceutical carrier can accumulate in tumor locus, but the medicine that is wrapped in carrier inside often can not in time discharge, and this makes therapeutic effect have a greatly reduced quality.
Stimuli responsive type (Stimuli-responsive) polymeric material is attracting the increasing sight of research worker (Jeong B in recent years, Gutowska A.Biotechnol.2002,20,305～311), this base polymer material can be discerned stimulus signal, makes response according to the power of signal and changes self conformation.Wherein, the oncotherapy research field of greatest concern be can the response environment pH value to change and change the polymeric material of dissolubility, this mainly is based on following reason: the environment pH value (6.15～7.4) around most tumor tissues is all than low (the Vaupel P of physiology pH value (7.0～7.4) around the normal structure, Kallinowski F, Okunieff P.CANCER RESEARCH 1989,49,6449～6465) pH value lower (5.0～6.5), and in endosome in the cell and the lysosome.Utilize these features,, just might be implemented in the tumor locus fixed point and discharge medicine if can design suitable pH sensitive polymer as anti-cancer medicament carrier.In the world in medicine controlled releasing and technical field of biological material, some more well-known seminar attention in the research of pH response type polymer all concentrates on the poly-weak base base polymer at present.Gast, people such as AP have reported the (N with polymethylacrylic acid-2-, the N-lignocaine) ethyl ester (PDEAEMA) is pH dependency (the Lee AS of synthetic and micellar structure of the block copolymer of hydrophobic segment, Butun V, Vamvakaki M, Armes SP, Pople JA, Gast AP.Macromolecules 2002,35,8540～8551); It is that the responsive segmental new polymers of pH is as pharmaceutical carrier (Lynn DM, Langer R.J.Am.Chem.Soc.2000,122,10761～10768) that Langer, R group have developed with μ-amino ester; Bae leader's seminar is then carrying out extensive studies work (Lee ES, Na K, BaeYH.Nano Lett., 2005,5,325～329) aspect polyhistidyl block copolymer and the administration micelle thereof.These pH sensitive polymers as pharmaceutical carrier almost are block copolymer, its synthetic living polymerization of mainly taking, condition is harsh and often need to use the heavy metal of suitable dosage to see as catalyst, and these problems make their medical applications be very restricted.
Summary of the invention
The invention provides a kind of is the pH response type amphipathic stem-grafting polyphosphazenes feeding micelle of vector construction nuclear with pH responsive type amphiphilic grafted polyphosphazene, and this administration micelle has the releasing effect of good targeting and medicine.
A kind of pH response type amphipathic stem-grafting polyphosphazenes feeding micelle contains pH response type amphiphilic grafted polyphosphazene and at least a hydrophobic drug;
In the gross weight of medicine and pH response type amphiphilic grafted polyphosphazene, the weight percentage of medicine is 0.001%～30%, and the weight percentage of pH response type amphiphilic grafted polyphosphazene is 70%～99.999%;
Described hydrophobic drug is hydrophobic anticancer drug or hydrophobicity chemotherapeutics, in the preferred paclitaxel of described hydrophobic anticancer drug, Docetaxel, amycin, daunorubicin, methotrexate, the ametycin one or more, most preferably one or both in paclitaxel, the amycin.
Described pH response type amphiphilic grafted polyphosphazene is a skeleton with the poly phosphazene segment with biological degradability, by grafting hydrophilic polymer segment on the poly phosphazene main chain, pH sensitive group and can reinforced polymeric material and the hydrophobic group of medicine affinity is final constitutes complete pH responsive type amphiphilic grafted polyphosphazene.
Described hydrophilic polymer generally can be selected the amino Polyethylene Glycol (NH of the end of good biocompatibility for use
2-PEG), its structural formula is as follows:
The micel that the optional usefulness of the responsive hydrophobic group of described pH has the tertiary amine structure, thus this quasi-molecule when pH value is lower than certain value, can take place protonated realization by hydrophobic to hydrophilic transformation.
The described hydrophobic group that is used for reinforcing material and medicine affinity can be selected biodegradable amino-acid ester for use, as N-(ethyl acetate base) amino, N-(ethyl benzoate base)-4-amino, N-[3-(β-indole) ethyl propionate base]-2-amino etc.
Described pH response type amphiphilic grafted polyphosphazene has the general structure shown in the formula (1):
In the formula, n=20～200;
R2 is selected from the pH sensitive group
In a kind of;
R3, R4 are identical or different, are selected from
In the quality of pH response type amphiphilic grafted polyphosphazene, the quality percentage composition of R1 is 2%～94.5%, and the quality percentage composition of R2 is 5%～97%, and the quality percentage composition of R3 is 0～90%, and the quality percentage composition of R4 is 0～90%.
For guaranteeing that the medicine micelle has the redispersibility of good drug loading, sustained release, stability, lyophilized powder and the timely degradation property of polymer, the number-average molecular weight of described pH response type amphiphilic grafted polyphosphazene is 4000～100000, preferred 5000～50000.
The high polymer water solublity is high more more for the content of R1 in the pH response type amphiphilic grafted polyphosphazene, and R2 content high polymer pH sensitive property more is strong more, and the adding of other group mainly is the affinity that improves micelle and medicine.
As preferably:
Described pH response type amphiphilic grafted polyphosphazene, in the quality of pH response type amphiphilic grafted polyphosphazene, the quality percentage composition of R1 is 60%～70%, and the quality percentage composition of R2 is 5%～39%, the quality percentage composition of R3 is 0～34.5%, and the quality percentage composition of R4 is 0～34.5%.
The molecular weight distribution of described pH response type amphiphilic grafted polyphosphazene is 1.5～4.0.
Described pH response type amphiphilic grafted polyphosphazene has the general structure shown in formula (2), formula (3) or the formula (4):
In the formula (2), n=20～200, R3, R4 are identical or different, are selected from
In the formula (3), n=20～200, R3 is selected from
In the formula (4), n=20～200.
The synthetic method of described pH response type amphiphilic grafted polyphosphazene comprises the steps:
(1) main chain polymerization: with chloro phosphine nitrile cyclic trimer ((PNCl
2)
3) 245 ℃～255 ℃ ring-opening polymerizations 3 hours～5 hours, make poly-(dichloro-phosphine nitrile);
(2) graft reaction: above-mentioned poly-(dichloro-phosphine nitrile) is dissolved in the oxolane, dropping contains oxolane (THF) solution of grafting with chemical compound and triethylamine (TEA), after reacting at least 8h, remove by filter insoluble matter, filtrate concentrates the back ether sedimentation, vacuum drying obtains the polymer of white powder, i.e. the pH response type amphiphilic grafted polyphosphazene;
Wherein, described grafting is selected N for use with chemical compound, the amino benzimidazole (ABI) of N-diisopropyl ethylenediamine (DPA), 1-butyl-4-(methylamino) piperidines (PPMA), 2-, 1-(3-aminopropyl) imidazoles (API), pantonine-imidazole radicals ethyl propionate (be the histidine ethyl ester, HiSE) in one or more and hold amino Polyethylene Glycol (NH
2-PEG);
Perhaps, described grafting is selected N for use with chemical compound, in N-diisopropyl ethylenediamine, 1-butyl-4-(methylamino) piperidines, the amino benzimidazole of 2-, 1-(3-aminopropyl) imidazoles, the pantonine-imidazole radicals ethyl propionate one or more, 4-benzocaine (EAB), ethyl aminoacetate (are glycine ethyl ester, EtGly), the beta-hydroxy alanine ethyl ester (be serine ethyl ester, SEE) in one or more and hold amino Polyethylene Glycol.
The number-average molecular weight of Polyethylene Glycol is preferably 350～5000, further preferred 1100～2000, preferred especially 2000 in the amino Polyethylene Glycol of described end.
I haven't seen you for ages causes reaction not exclusively because the triethylamine amount is crossed, and crosses to cause polymer precipitation at most, and therefore, grafting is preferably 1: 1 with the mol ratio of chemical compound and triethylamine.
Oxolane is the very big organic solvent of soluble end, solvable depolymerization (dichloro-phosphine nitrile) and various grafting chemical compound, and as dissolvant of reaction system, its consumption does not have much affect to graft reaction, thereby does not do concrete qualification.
Process of grafting of the present invention must be carried out in highly anhydrous environment, there is no particular limitation for reaction temperature, for example room temperature or room temperature all can, for the synthetic bigger polymer of substituent group steric hindrance, in synthetic reaction process, generally need carry out heating reflux reaction, be more conducive to the carrying out that reacts.
According to the substituent group on the pH response type amphiphilic grafted polyphosphazene of design in advance, described graft compound preferably drips according to predetermined order.Generally, can drip the bigger substituent group of steric hindrance earlier, drip the less substituent group of steric hindrance again according to the substituent steric hindrance size of pre-adding for the various substituent groups of design in advance of grafting more easily on poly-(dichloro-phosphine nitrile).
The polymer of described white powder carries out purification process, its purification process comprises: the polymer of white powder is dialysed in water, to remove unreacted 2-(Polyethylene Glycol amino) ethylamino, can adopt bag filter to dialyse, collect product through lyophilization again.
With N, the N-diisopropyl ethylenediamine is an example, N, the structural formula of N-diisopropyl ethylenediamine be suc as formula shown in a, slough a hydrogen on its primary amine after, be grafted on the P atom of poly phosphazene main chain, become pH sensitive group (being the grafting group): N-[2-(N ', N '-diisopropylaminoethyl) ethyl] amino, its structural formula is suc as formula b, this group only is at N, sloughs a hydrogen on the primary amine of N-diisopropyl ethylenediamine.
All the other pH sensitive groups also only are to slough a hydrogen evolution on the primary amine of 1-butyl-4-(methylamino) piperidines, the amino benzimidazole of 2-, 1-(3-aminopropyl) imidazoles, pantonine-imidazole radicals ethyl propionate, and its structural formula is followed successively by:
Other grafting also only is amino, the N-[ethylene lactic acid ethoxycarbonyl of N-(ethyl benzoate base)-4-amino, N-(ethyl acetate base) of sloughing a hydrogen evolution on the primary amine of ethylaminobenzoate, glycine ethyl ester, serine ethyl ester with the group of chemical compound] amino, its structural formula is followed successively by:
The micellar particle diameter of described pH response type amphipathic stem-grafting polyphosphazenes feeding is 10 nanometers～300 nanometers.
Described pH response type amphipathic stem-grafting polyphosphazenes feeding micelle can be by thin film aquation method or dialysis preparation, and micelle can be to exist with aqueous dispersions or lyophilized powder state, and the preparation method of its aqueous dispersions may further comprise the steps:
Be dissolved in hydrophobic drug and pH response type amphiphilic grafted polyphosphazene in the organic solvent jointly, form the solution of mixture, with the heating or the decompression mode remove organic solvent after, form the homogeneous film of polymer and medicine, add the buffer salt solution aquation dissolving films of pure water or pH value 7.0～7.4, obtain pH responding type polyphosphazene administration micellar solution.
In described organic solvent particular methanol, acetone, oxolane, the dichloromethane one or more.
Described buffer salt solution selects for use this area buffer salt solution commonly used to get final product.
The concentration of pH response type amphiphilic grafted polyphosphazene is 1mg/ml～2g/ml in the solution of described mixture.
The concentration expressed in percentage by weight of described pH responding type polyphosphazene administration micellar solution is 1%～50%.
Described pH responding type polyphosphazene administration micellar solution can make pH responding type polyphosphazene administration micelle freeze-drying powder through lyophilization.This lyophilized powder can be scattered in the water certainly, forms the micellar aqueous dispersions of polymeric medicine, and the micelle size is generally in 10 nanometers～300 nanometers.
With the amycin is example, the described pH responding type polyphosphazene administration micelle that contains amycin, preferably, amycin weight be amycin and pH response type amphiphilic grafted polyphosphazene gross weight 2%～30%, the Polyethylene Glycol quality accounts for more than 40% of total polymer mass in the pH response type amphiphilic grafted polyphosphazene.
The present invention has following advantage:
1) hydrophobic group can effectively wrap by hydrophobic interaction and carry the cancer therapy drug that is insoluble in water in the amphipathic graft copolymer;
2) the drug-carrying polymer micelle lyophilized powder of above-mentioned preparation is suitable for drug administration by injection, and has and be easy to store, transport, use advantage easily, can be according to patient's needs, and the water that employing contains heterogeneity disperses.
3) this drug delivery system most important characteristic is to engulf when entering endosome and lysosome around pharmaceutical carrier arrives tumor tissues or by tumor cell, because the environment pH value reduces, the pH response group generation that originally is in micelle hydrophobic inner core place is protonated, causes micelle to disintegrate and will wrap the medicine that carries discharging in a large number at short notice thereby destroyed the micellar stability of original administration.By choosing suitable pH sensitive group and controlling the pH value that its substitution value in graft polymers in theory can telomerized polymer responds, this makes this drug delivery system can have drug release characteristic very flexibly: what carry when the system bag is during at the antibody of tumor cell surface or memebrane protein inhibitor, the polymer that can select to respond tumor tissues peripheral environment pH value is as carrier, and what carry when bag is that the polymer of then selecting to respond low pH value in the time of need entering the medicine that cytoplasm or nucleus work is made carrier.In addition, if connect tumor targeted molecular in the hydrophilic chain outer end then can further improve the transport of drug effect of carrier.Based on above characteristics, this administration micelle volume is expected to increase substantially therapeutic effect, the reversal of multidrug resistance of tumor cells of antitumor drug and can effectively reduces toxic and side effects.
4) because therefore the degradation characteristic of pH response type amphiphilic grafted polyphosphazene and depend primarily on its hydrophobic substituent with the affinity of medicine, just can regulate micelle degradation feature and medicine carrying performance by adding suitable hydrophobic substituent.Simultaneously, can be relatively easy to control micellar drug loading and size by method and the parameter that changes in the medicine carrying process, thereby can more easily satisfy the requirement of different pharmaceutical and preparation.
5) the micellar drug release of administration of the present invention has the pH response, can fixed point ground near tumor tissues or in the tumor cell rapid delivery of pharmaceuticals have the function of reversal of multidrug resistance of tumor cells, the good biocompatibility toxicity of this amphipathy polyphosphazene graft copolymer is minimum simultaneously, is suitable as the antitumor drug targeting vector.
Description of drawings
The pH responding type polyphosphazene that Fig. 1, Fig. 2 are different polymer manufacture carries the micellar vitro drug release curve of amycin;
Wherein: Fig. 1-1 represents the P1 carrier micelle of embodiment 6 preparations in the release profiles under the pH=7.4 condition;
Fig. 1-2 represents the P2 carrier micelle of embodiment 7 preparations in the release profiles under the pH=7.4 condition;
Fig. 1-3 represents the P3 carrier micelle of embodiment 8 preparations in the release profiles under the pH=7.4 condition;
Fig. 1-4 represents the P1 carrier micelle of embodiment 6 preparations in the release profiles under the pH=5.5 condition;
Fig. 1-5 represents the P2 carrier micelle of embodiment 7 preparations in the release profiles under the pH=5.5 condition;
Fig. 1-6 represents the P3 carrier micelle of embodiment 8 preparations in the release profiles under the pH=5.5 condition;
Fig. 2-7 represents the P4 carrier micelle of embodiment 9 preparations in the release profiles under the pH=7.4 condition;
Fig. 2-8 represents the P5 carrier micelle of embodiment 10 preparations in the release profiles under the pH=7.4 condition;
Fig. 2-9 represents the P4 carrier micelle of embodiment 9 preparations in the release profiles under the pH=5.5 condition;
Fig. 2-10 represents the P5 carrier micelle of embodiment 10 preparations in the release profiles under the pH=5.5 condition.
Fig. 3 is several proton nmr spectras with pH response type amphiphilic grafted polyphosphazene of typical chemical constitution of the present invention;
Wherein, spectrogram 3-1 is the proton nmr spectra of the pH response type amphiphilic grafted polyphosphazene of embodiment 1 preparation;
Spectrogram 3-2 is the proton nmr spectra of the pH response type amphiphilic grafted polyphosphazene of embodiment 2 preparations;
Spectrogram 3-3 is the proton nmr spectra of the pH response type amphiphilic grafted polyphosphazene of embodiment 3 preparations;
Spectrogram 3-4 is the proton nmr spectra of the pH response type amphiphilic grafted polyphosphazene of embodiment 4 preparations;
Spectrogram 3-5 is the proton nmr spectra of the pH response type amphiphilic grafted polyphosphazene of embodiment 5 preparations;
Fig. 4 is the pH titration curve of pH response type amphiphilic grafted polyphosphazene;
Wherein, Fig. 4-1 is the pH titration curve of the pH response type amphiphilic grafted polyphosphazene of embodiment 1 preparation;
Fig. 4-2 is the pH titration curve (adopting the acid potentiometric titration) of the pH response type amphiphilic grafted polyphosphazene of embodiment 2 preparations.
The specific embodiment
(1) with the aluminum chloride is Preparation of Catalyst poly-(dichloro-phosphine nitrile)
Take by weighing 4g through the chloro phosphine nitrile cyclic trimer of sublimation purification and 0.2g aluminum trichloride (anhydrous) in the polymerization pipe of strict cleaning-drying in advance, evacuation and tube sealing, polyreaction is carried out 5h at 250 ℃, when the viscosity of question response thing is almost constant, stops polymerization and takes out the polymerization pipe cooling.The Kaifeng polymerization pipe adds an amount of dry toluene solution solubilizing reaction thing.The dissolving back uses petroleum ether precipitation, vacuum drying to obtain white elastomer, i.e. poly-(dichloro-phosphine nitrile).
(2) pass through the progressively synthetic pH response type amphiphilic grafted polyphosphazene copolymer of nucleophilic substitution
Above-mentioned linear poly-(the dichloro-phosphine nitrile) that makes of 0.5g is dissolved among the 20ml THF as reaction system, contains 2.2g NH to wherein slowly dripping
2The THF solution 50ml of-PEG (molecular weight is 2000) and 0.2ml TEA drips back normal-temperature reaction 8h.And then contain 1.1g N, the THF solution 10ml of N-diisopropyl ethylenediamine (DPA) and 1.0ml TEA to wherein dripping.Room temperature reaction 12h after-filtration is removed insoluble matter under the magnetic agitation, and filtrate concentrates the back and uses ether sedimentation, and vacuum drying obtains the polymer of micro-yellow powder shape.
Utilize bag filter (its molecular cut off (MWCO)=14000) that resulting polymers was dialysed in water two days, to remove unreacted NH
2-PEG
2000And through lyophilization collection product, obtain pH response type amphiphilic grafted polyphosphazene copolymer p 1, the quality percentage composition of the amino Polyethylene Glycol of hydrophilic section end is 65% in this copolymer, N-[2-(N ', N '-diisopropylaminoethyl) ethyl] amino quality percentage composition is 31%, its number-average molecular weight is 5000, molecular weight distribution 2.8, pKa value are 6.30.The nuclear magnetic spectrogram of this copolymer as shown in Figure 3, its structural formula is as follows:
Wherein, n=20～200, R3, R4 are identical or different, are selected from
(1) preparation of poly-(dichloro-phosphine nitrile) is with embodiment 1.
(2) contain 2.2g NH except in reaction system, slowly dripping successively
2The THF solution 50ml of-PEG (molecular weight is 2000) and 0.2ml TEA, contain 0.5g N, the THF solution 10ml of N-diisopropyl ethylenediamine (DPA) and 0.5ml TEA and containing outside 10 milliliters of the THF solution of 0.7g 4-benzocaine (EAB) and 0.5ml TEA, all the other operations are all with the step among the embodiment 1 (2), obtain pH response type amphiphilic grafted polyphosphazene copolymer p 2, the quality percentage composition of the amino Polyethylene Glycol of hydrophilic section end is 60% in this copolymer, N-[2-(N ', N '-diisopropylaminoethyl) ethyl] amino quality percentage composition is 15%, the quality percentage composition of N-(ethyl benzoate base)-4-amino is 20%, its number-average molecular weight is 5000, molecular weight distribution 2.8, pKa value are 6.34.The nuclear magnetic spectrogram of this copolymer as shown in Figure 3, its structural formula is as follows:
In the formula, n=20～200, R3 is selected from
(1) preparation of poly-(dichloro-phosphine nitrile) is with embodiment 1.
(2) contain 6g NH except in reaction system, slowly dripping successively
2The THF solution 50ml of-PEG (molecular weight is 2000) and 0.6ml TEA, contain 0.8g N, the THF solution 10ml of N-diisopropyl ethylenediamine (DPA) and 0.8ml TEA, 10 milliliters of THF solution that contain 10 milliliters of the THF solution of 0.8g 4-benzocaine (EAB) and 0.7ml TEA and 2.0g beta-hydroxy alanine ethyl ester (SEE) and 1.5ml TEA, and drip behind the class raw material outside the normal-temperature reaction 8h at every turn, all the other operations are all with the step among the embodiment 1 (2), obtain pH response type amphiphilic grafted polyphosphazene copolymer p 3, the quality percentage composition of the amino Polyethylene Glycol of hydrophilic section end is 70% in this copolymer, N-[2-(N ', N '-diisopropylaminoethyl) ethyl] amino quality percentage composition is 10%, the quality percentage composition of N-(ethyl benzoate base)-4-amino is 6%, N-[ethylene lactic acid ethoxycarbonyl] amino quality percentage composition is 4%, its number-average molecular weight is 6600, molecular weight distribution 3.0, pKa value are 6.50.The nuclear magnetic spectrogram of this copolymer as shown in Figure 3, its structural formula is as follows:
In the formula, n=20～200.
Embodiment 4 pH response type amphiphilic grafted polyphosphazene synthetic methods
(1) preparation of poly-(dichloro-phosphine nitrile) is with embodiment 1.
(2) contain 4g NH except in reaction system, slowly dripping successively
2The THF solution 50ml of-PEG (molecular weight is 2000) and 0.4ml TEA and contain 1.0g N, the THF solution 10ml of N-diisopropyl ethylenediamine (DPA) and 1.0ml TEA, and drip behind the class raw material outside the normal-temperature reaction 12h at every turn, all the other operations are all with the step among the embodiment 1 (2), obtain pH response type amphiphilic grafted polyphosphazene copolymer p 4, the quality percentage composition of the amino Polyethylene Glycol of hydrophilic section end is 80% in this copolymer, N-[2-(N ', N '-diisopropylaminoethyl) ethyl] amino quality percentage composition is 15%, its number-average molecular weight is 11000, molecular weight distribution 2.0, pKa value are 6.32.The nuclear magnetic spectrogram of this copolymer as shown in Figure 3, its structural formula is as follows:
Wherein, n=20～200, R3, R4 are identical or different, are selected from
(1) preparation of poly-(dichloro-phosphine nitrile) is with embodiment 1.
(2) contain 2.2g NH except in reaction system, slowly dripping successively
2The THF solution 50ml of-PEG (molecular weight is 2000) and 0.2ml TEA, contain 0.2g N, the THF solution 10ml of N-diisopropyl ethylenediamine (DPA) and 0.2ml TEA and the THF solution 10ml that contains 1.1g 4-benzocaine (EAB) and 0.9ml TEA, and drip behind the class raw material outside the normal-temperature reaction 10h at every turn, all the other operations are all with the step among the embodiment 1 (2), obtain pH response type amphiphilic grafted polyphosphazene copolymer p 5, the quality percentage composition of the amino Polyethylene Glycol of hydrophilic section end is 64% in this copolymer, N-[2-(N ', N '-diisopropylaminoethyl) ethyl] amino quality percentage composition is 5%, the quality percentage composition of N-(ethyl benzoate base)-4-amino is 30%, its number-average molecular weight is 10000, molecular weight distribution 3.0, pKa value are 6.40.The nuclear magnetic spectrogram of this copolymer as shown in Figure 3, its structural formula is as follows:
In the formula, n=20～200, R3 is selected from
The micellar preparation of embodiment 6 administrations
(1) polymer drug-carried micellar preparation of P1 and sign
The pH response type amphiphilic grafted polyphosphazene P1 of 30mg embodiment 1 preparation is dissolved in the 20mL dichloromethane with the 6mg amycin, pack into and volatilize organic solvent with Rotary Evaporators in 40 degrees centigrade of reduced pressure in the eggplant-shape bottle, obtain the homogeneous film of medicine and copolymer, add the 10ml pure water with Film Fractionation and with 0.45 μ m membrane filtration, obtain the aqueous dispersions that clear liquid is pH response type amphiphilic grafted polyphosphazene P1 amycin administration micelle (P1-DOX), this aqueous dispersions lyophilization gets the pH responding type polyphosphazene and carries amycin micelle freeze-drying powder.With the micellar particle diameter of dynamic light scattering determination, the micellar character of resulting medicine sees Table 2.
(2) polymer drug-carried micelle (P1-DOX) extracorporeal releasing experiment
The pH responding type polyphosphazene that takes by weighing above-mentioned preparation carries amycin micelle freeze-drying powder 5mg, phosphate buffer with 10ml pH=7.4 or pH=5.5 in bag filter dissolves, place 20mlpH=7.4 or pH=5.5 phosphate buffer to carry out extracorporeal releasing experiment then, stir.Phosphate buffer outside certain hour is got bag filter, analyze wherein medicament contg with ultraviolet absorption spectroscopy, replenish the fresh phosphoric salt buffer of equivalent simultaneously, the result can see that the drug release rate of carrier micelle in low ph environment (pH=6.0～4.0) improves greatly as shown in Figure 1.
(3) cytotoxicity of polymer drug-carried micelle (P1-DOX)
The present invention is a model cell with breast carcinoma mdr cell (MCF7/ADR), estimates not the pH response type amphiphilic grafted polyphosphazene P1 micelle (P1 polymer micelle) of medicine carrying and the cytotoxicity of P1-DOX.
Concrete grammar is: with breast carcinoma mdr cell (MCF7/ADR cell) is model cell, and in 96 porocyte culture plates, every hole adds 100 μ l and contains 1 * 10
5The culture fluid of individual MCF7/ADR cell places 5% (percent by volume) CO
2, cultivated in 37 ℃ of incubators 24 hours, treat that cell is adherent fully after, add P1 polymer micelle solution, P1-DOX solution and the free amycin aqueous solution of variable concentrations in the cell, be contrast with undressed blank cell, 4 multiple holes are established in every hole.Normal RMPI16401640 culture fluid continues to cultivate 24 hours, and every hole adds the 3-(4,5-dimethylthiazole-2)-2 of 5mg/mL, and 5-diphenyl tetrazole bromine salt (MTT) 31.5 μ l put 5%CO
2, cultivate 4 hours abandoning supernatant in 37 ℃ of incubators, every hole adds 200 μ l dimethyl sulphoxide solutions, measures light absorption value with microplate reader, calculates cell inhibitory rate.The cell inhibitory rate computing formula is as follows: cell inhibitory rate (%)=(blank group light absorption value-experimental group light absorption value)/blank group light absorption value * 100%
After measured, the P1 polymer micelle is very little to the cytotoxicity of two kinds of cells, is that the survival rate of cell is still more than 85% after 48 hours effect under the condition of 1mg/ml at the P1 polymer concentration, and cytotoxicity is 0 grade, has excellent biological compatibility.Medicine carrying amycin polymer micelle (P1-DOX) solution and free amycin are to the IC of MCF7/ADR cell
50See Table 3, the result shows that polymer micelle can produce the amycin drug resistance inversion effect of MCF7/ADR.
The character of the drug-carrying polymer micelle of the different polymer manufacture of table 2
| Polymer | Numbering | Mean diameter (nm) | Drug loading (%) | Envelop rate (%) |
| ??P1 | ??P1-DOX | ??192 | ??10.2 | ??63 |
| ??P2 | ??P2-DOX | ??280 | ??12.6 | ??76 |
| ??P3 | ??P3-DOX | ??290 | ??12.7 | ??77 |
| ??P4 | ??P4-DOX | ??271 | ??16.1 | ??97 |
| ??P5 | ??P5-DOX | ??113 | ??16.0 | ??96 |
Table 3
| Prescription | ??IC 50(μg/ml)??MCF7/ADR |
| Free amycin aqueous solution | ??8.45 |
| ??P1-DOX | ??0.63 |
The micellar preparation of embodiment 7 administrations
(1) polymer drug-carried micellar preparation of P2 and sign, except the pH response type amphiphilic grafted polyphosphazene P2 that adopts embodiment 2 preparations, all the other are operated with the step among the embodiment 6 (1), and the micellar character of resulting medicine sees Table 2.
(2) the polymer drug-carried micelle of P2 (P2-DOX) extracorporeal releasing experiment, operation is with the step among the embodiment 6 (2), and the result can see that the drug release rate of carrier micelle in low ph environment (pH=6.0～4.0) improves greatly as shown in Figure 1.
(3) cytotoxicity of polymer drug-carried micelle (P2-DOX)
Method is with the step among the embodiment 6 (3).
After measured, the P2 polymer micelle is very little to the cytotoxicity of two kinds of cells, is that the survival rate of cell is still more than 100% after 48 hours effect under the condition of 1mg/ml at the P2 polymer concentration, and cytotoxicity is 0 grade, has excellent biological compatibility.Medicine carrying amycin polymer micelle solution (P2-DOX) and free amycin are to the IC of MCF7/ADR cell
50See Table 4, the result shows that polymer micelle can produce the amycin drug resistance inversion effect of MCF7/ADR.
Table 4
| Prescription | ??IC 50(μg/ml)??MCF7/ADR |
| Free amycin solution | ??8.45 |
| ??P2-DOX | ??0.13 |
The micellar preparation of embodiment 8 administrations
(1) polymer drug-carried micellar preparation of P3 and sign, except the pH response type amphiphilic grafted polyphosphazene P3 that adopts embodiment 3 preparations, all the other are operated with the step among the embodiment 6 (1), and the micellar character of resulting medicine sees Table 2.
(2) the polymer drug-carried micelle of P3 (P3-DOX) extracorporeal releasing experiment, operation is with the step among the embodiment 6 (2), and the result can see that the drug release rate of carrier micelle in low ph environment (pH=6.0～4.0) improves greatly as shown in Figure 1.
(3) cytotoxicity of polymer drug-carried micelle (P3-DOX)
Method is with the step among the embodiment 6 (3).
After measured, the P3 polymer micelle is very little to the cytotoxicity of two kinds of cells, is that the survival rate of cell is still more than 90% after 48 hours effect under the condition of 1mg/ml at the P3 polymer concentration, and cytotoxicity is 0 grade, has excellent biological compatibility.Medicine carrying amycin polymer micelle solution (P3-DOX) and free amycin are to the IC of MCF7/ADR cell
50See Table 5, the result shows that polymer micelle can produce the amycin drug resistance inversion effect of MCF7/ADR.
Table 5
| Prescription | ??IC 50(μg/ml)??MCF7/ADR |
| Free amycin solution | ??8.45 |
| ??P3-DOX | ??0.85 |
The micellar preparation of embodiment 9 administrations
(1) polymer drug-carried micellar preparation of P4 and sign, except the pH response type amphiphilic grafted polyphosphazene P4 that adopts embodiment 4 preparations, all the other are operated with the step among the embodiment 6 (1), and the micellar character of resulting medicine sees Table 2.
(2) the polymer drug-carried micelle of P4 (P4-DOX) extracorporeal releasing experiment, operation is with the step among the embodiment 6 (2), and the result can see that the drug release rate of carrier micelle in low ph environment (pH=6.0～4.0) improves greatly as shown in Figure 2.
(3) cytotoxicity of polymer drug-carried micelle (P4-DOX)
Method is with the step among the embodiment 6 (3).
After measured, the P4 polymer micelle is very little to the cytotoxicity of two kinds of cells, is that the survival rate of cell is still more than 95% after 48 hours effect under the condition of 1mg/ml at the P4 polymer concentration, and cytotoxicity is 0 grade, has excellent biological compatibility.Medicine carrying amycin polymer micelle solution (P4-DOX) and free amycin are to the IC of MCF7/ADR cell
50See Table 5, the result shows that polymer micelle can produce the amycin drug resistance inversion effect of MCF7/ADR.
Table 5
| Prescription | ??IC 50(μg/ml)??MCF7/ADR |
| Free amycin solution | ??8.45 |
| ??P4-DOX | ??0.70 |
The micellar preparation of embodiment 10 administrations
(1) polymer drug-carried micellar preparation of P5 and sign, except the pH response type amphiphilic grafted polyphosphazene P5 that adopts embodiment 5 preparations, all the other are operated with the step among the embodiment 6 (1), and the micellar character of resulting medicine sees Table 2.
(2) the polymer drug-carried micelle of P5 (P3-DOX) extracorporeal releasing experiment, operation is with the step among the embodiment 6 (2), and the result can see that the drug release rate of carrier micelle in low ph environment (pH=6.0～4.0) improves greatly as shown in Figure 2.
(3) cytotoxicity of polymer drug-carried micelle (P5-DOX)
Method is with the step among the embodiment 6 (3).
After measured, the P5 polymer micelle is very little to the cytotoxicity of two kinds of cells, is that the survival rate of cell is still more than 88% after 48 hours effect under the condition of 1mg/ml at the P5 polymer concentration, and cytotoxicity is 0 grade, has excellent biological compatibility.Medicine carrying amycin polymer micelle solution (P5-DOX) and free amycin are to the IC of MCF7/ADR cell
50See Table 5, the result shows that polymer micelle can produce the amycin drug resistance inversion effect of MCF7/ADR.
Table 5
| Prescription | ??IC 50(μg/ml)??MCF7/ADR |
| Free amycin solution | ??8.45 |
| ??P5-DOX | ??0.40 |
Need not further to elaborate, believe and adopt the disclosed content in front, those skilled in the art can use to greatest extent, and these equivalent form of values fall within the application's appended claims institute restricted portion equally.In addition, the preferred specific embodiments of front should be understood that only to illustrate, but not limits the scope of the invention by any way.
Claims (10)
1. a pH response type amphipathic stem-grafting polyphosphazenes feeding micelle is characterized in that, contains pH response type amphiphilic grafted polyphosphazene and at least a hydrophobic drug in the described administration micelle;
In the gross weight of medicine and pH response type amphiphilic grafted polyphosphazene, the weight percentage of medicine is 0.001%～30%, and the weight percentage of pH response type amphiphilic grafted polyphosphazene is 70%～99.999%;
Described hydrophobic drug is hydrophobic anticancer drug or hydrophobicity chemotherapeutics;
Described pH response type amphiphilic grafted polyphosphazene has the general structure shown in the formula (1):
Wherein, R1, R2, R3 and R4 are the grafting groups, and R1 is selected from
In the formula, n=20～200;
R2 is selected from
In a kind of;
R3, R4 are identical or different, are selected from
In a kind of.
2. pH response type amphipathic stem-grafting polyphosphazenes feeding micelle as claimed in claim 1, it is characterized in that, quality in the pH response type amphiphilic grafted polyphosphazene, the quality percentage composition of R1 is 2%～94.5%, the quality percentage composition of R2 is 5%～97%, the quality percentage composition of R3 is 0～90%, and the quality percentage composition of R4 is 0～90%.
3. pH response type amphipathic stem-grafting polyphosphazenes feeding micelle as claimed in claim 2, it is characterized in that, quality in the pH response type amphiphilic grafted polyphosphazene, the quality percentage composition of R1 is 60%～70% in the described pH response type amphiphilic grafted polyphosphazene, the quality percentage composition of R2 is 5%～39%, the quality percentage composition of R3 is 0～34.5%, and the quality percentage composition of R4 is 0～34.5%.
4. pH response type amphipathic stem-grafting polyphosphazenes feeding micelle as claimed in claim 1 is characterized in that the number-average molecular weight of described pH response type amphiphilic grafted polyphosphazene is 4000～100000;
Perhaps, the molecular weight distribution of described pH response type amphiphilic grafted polyphosphazene is 1.5～4.0.
5. pH response type amphipathic stem-grafting polyphosphazenes feeding micelle as claimed in claim 1 is characterized in that, described pH response type amphiphilic grafted polyphosphazene has the general structure shown in formula (2), formula (3) or the formula (4):
In the formula (2), n=20～200, R3, R4 are identical or different, are selected from
(3)
In the formula (3), n=20～200, R3 is selected from
In the formula (4), n=20～200.
6. pH response type amphipathic stem-grafting polyphosphazenes feeding micelle as claimed in claim 1, it is characterized in that described hydrophobic anticancer drug is selected one or more in paclitaxel, Docetaxel, amycin, daunorubicin, methotrexate, the ametycin for use.
7. pH response type amphipathic stem-grafting polyphosphazenes feeding micelle as claimed in claim 1 is characterized in that, the micellar particle diameter of described pH response type amphipathic stem-grafting polyphosphazenes feeding is 10 nanometers～300 nanometers.
8. the micellar preparation method of pH response type amphipathic stem-grafting polyphosphazenes feeding as claimed in claim 1 may further comprise the steps:
Be dissolved in hydrophobic drug and pH response type amphiphilic grafted polyphosphazene in the organic solvent jointly, form the solution of mixture, with the heating or the decompression mode remove organic solvent after, form the homogeneous film of polymer and medicine, adding pure water or pH value are 7.0～7.4 buffer salt solution aquation dissolving films, obtain pH responding type polyphosphazene administration micellar solution.
9. preparation method as claimed in claim 8 is characterized in that described organic solvent is selected one or more in methanol, acetone, oxolane, the dichloromethane for use;
Perhaps, the concentration of pH response type amphiphilic grafted polyphosphazene is 1mg/m1～2g/ml in the solution of described mixture;
Perhaps, the concentration expressed in percentage by weight of described pH responding type polyphosphazene administration micellar solution is 1%～50%.
10. preparation method as claimed in claim 8 is characterized in that, described pH responding type polyphosphazene administration micellar solution makes pH responding type polyphosphazene administration micelle freeze-drying powder through lyophilization.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101180323A CN101785865B (en) | 2010-03-04 | 2010-03-04 | pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101180323A CN101785865B (en) | 2010-03-04 | 2010-03-04 | pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101785865A true CN101785865A (en) | 2010-07-28 |
| CN101785865B CN101785865B (en) | 2012-02-15 |
Family
ID=42529334
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101180323A Expired - Fee Related CN101785865B (en) | 2010-03-04 | 2010-03-04 | pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101785865B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101984956A (en) * | 2010-11-03 | 2011-03-16 | 浙江大学 | Application of pH sensitive type amphiphilic graft polyphosphazene for preparing administration vesicle |
| CN106065079A (en) * | 2016-06-08 | 2016-11-02 | 江苏华昌织物有限公司 | A kind of preparation method of the Synthesis, Characterization of Polyphosphazenes for medicament slow release |
| CN108478526A (en) * | 2018-04-03 | 2018-09-04 | 延边大学 | Double-response amphipathy polyphosphazene target medicine carrier material and preparation method thereof |
| CN111248195A (en) * | 2020-02-24 | 2020-06-09 | 暨南大学 | Nano-silver-loaded polyphosphazene composite antibacterial agent and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001007505A1 (en) * | 1999-07-21 | 2001-02-01 | The Penn State Research Foundation | Polyphosphazene polymers |
| CN1613890A (en) * | 2003-11-04 | 2005-05-11 | 北京化工大学 | Biological degradable PES graft polyphosphonitrile copolymer and its preparation |
| WO2006113274A1 (en) * | 2005-04-15 | 2006-10-26 | Parallel Solutions, Inc. | Biodegradable polyphosphazenes containing pyrrolidone side groups |
| WO2009037313A1 (en) * | 2007-09-18 | 2009-03-26 | Nirvana's Tree House B.V. | Amphiphilic copolymers and compositions containing such polymers |
-
2010
- 2010-03-04 CN CN2010101180323A patent/CN101785865B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001007505A1 (en) * | 1999-07-21 | 2001-02-01 | The Penn State Research Foundation | Polyphosphazene polymers |
| CN1613890A (en) * | 2003-11-04 | 2005-05-11 | 北京化工大学 | Biological degradable PES graft polyphosphonitrile copolymer and its preparation |
| WO2006113274A1 (en) * | 2005-04-15 | 2006-10-26 | Parallel Solutions, Inc. | Biodegradable polyphosphazenes containing pyrrolidone side groups |
| WO2009037313A1 (en) * | 2007-09-18 | 2009-03-26 | Nirvana's Tree House B.V. | Amphiphilic copolymers and compositions containing such polymers |
Non-Patent Citations (3)
| Title |
|---|
| 《"以岭医药杯"第八届全国青年药学工作者最新科研成果交流会论文集》 20061231 张建祥等 基于两亲聚膦腈的聚合物胶束的合成及其初步体外评价 248-254 1-10 , 2 * |
| 《pH响应型聚膦腈水凝胶的制备及其应用研究》 20081130 洪挺 pH响应型聚膦腈水凝胶的制备及其应用研究 48-49 5 , 2 * |
| 《以聚乙二醇为亲水链的两亲接枝聚膦腈的自组装及其药物传递系统》 20091130 郑程 以聚乙二醇为亲水链的两亲接枝聚膦腈的自组装及其药物传递系统 26-28,53-54 5 , 2 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101984956A (en) * | 2010-11-03 | 2011-03-16 | 浙江大学 | Application of pH sensitive type amphiphilic graft polyphosphazene for preparing administration vesicle |
| CN106065079A (en) * | 2016-06-08 | 2016-11-02 | 江苏华昌织物有限公司 | A kind of preparation method of the Synthesis, Characterization of Polyphosphazenes for medicament slow release |
| CN108478526A (en) * | 2018-04-03 | 2018-09-04 | 延边大学 | Double-response amphipathy polyphosphazene target medicine carrier material and preparation method thereof |
| CN108478526B (en) * | 2018-04-03 | 2020-02-14 | 延边大学 | Dual-response amphiphilic polyphosphazene targeted drug carrier material and preparation method thereof |
| CN111248195A (en) * | 2020-02-24 | 2020-06-09 | 暨南大学 | Nano-silver-loaded polyphosphazene composite antibacterial agent and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101785865B (en) | 2012-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lin et al. | Amphiphilic miktoarm star copolymer (PCL) 3-(PDEAEMA-b-PPEGMA) 3 as pH-sensitive micelles in the delivery of anticancer drug | |
| Gao et al. | Preparation and characterization of Pluronic/TPGS mixed micelles for solubilization of camptothecin | |
| Mukerabigwi et al. | Polymersome nanoreactors with tumor pH-triggered selective membrane permeability for prodrug delivery, activation, and combined oxidation-chemotherapy | |
| CN102037058B (en) | Docetaxel polymer derivative, method for producing same and use of same | |
| Bersani et al. | pH-sensitive stearoyl-PEG-poly (methacryloyl sulfadimethoxine) decorated liposomes for the delivery of gemcitabine to cancer cells | |
| Wang et al. | Preparation of tacrolimus loaded micelles based on poly (ɛ-caprolactone)–poly (ethylene glycol)–poly (ɛ-caprolactone) | |
| CN101785865B (en) | pH response type amphipathic stem-grafting polyphosphazenes feeding micelle and preparation method thereof | |
| CN108670954B (en) | Chemotherapeutic drug co-loaded glycyrrhetinic acid prodrug micelle and preparation method thereof | |
| CN111632153B (en) | Chemical gene drug co-loaded targeting nano drug delivery system and preparation method thereof | |
| Xiao et al. | Co-delivery of metformin and paclitaxel via folate-modified pH-sensitive micelles for enhanced anti-tumor efficacy | |
| CN104490786B (en) | Preparation method and application of targeted multi-function double drug-loading liposome | |
| CN113683541B (en) | Disulfiram-based amphiphilic block copolymer prodrug and preparation method and application thereof | |
| CN105796482A (en) | Paclitaxel slow-release temperature-sensitive gel and preparation method thereof | |
| CN105535991A (en) | Amphiphilic basic group conjugate nanometer particles for treating tumors and preparation method thereof | |
| CN111333786B (en) | Preparation method of acid-sensitive adriamycin prodrug based on zwitterion and folic acid targeting | |
| CN107998081A (en) | A kind of application for targeting reduction response vesica Nano medication in treatment of brain tumor medicine is prepared | |
| CN101759855B (en) | pH response type amphiphilic grafted polyphosphazene and synthesis method thereof | |
| CN101984956A (en) | Application of pH sensitive type amphiphilic graft polyphosphazene for preparing administration vesicle | |
| CN109481400A (en) | A kind of Liver targeting adriamycin/Bcl-2 siRNA carries nano-micelle and its preparation method and application altogether | |
| CN107496936A (en) | A kind of both sexes small molecule self assembly targeted nanoparticles drug-loading system and preparation method thereof | |
| CN104546708A (en) | Taxol micelle preparation | |
| CN103446053A (en) | Targeting liposome drug delivery system for multidrug resistant tumors | |
| US9339549B2 (en) | Cationic graft-copolymer for drug delivery system | |
| CN107674196B (en) | Docetaxel prodrug with anti-tumor effect and preparation method thereof | |
| CN110200914A (en) | A kind of taxol micella preparation process and product |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120215 Termination date: 20210304 |