CN109833295A - A kind of polymer micelle and preparation method thereof of cancer target and pH response - Google Patents
A kind of polymer micelle and preparation method thereof of cancer target and pH response Download PDFInfo
- Publication number
- CN109833295A CN109833295A CN201811248768.5A CN201811248768A CN109833295A CN 109833295 A CN109833295 A CN 109833295A CN 201811248768 A CN201811248768 A CN 201811248768A CN 109833295 A CN109833295 A CN 109833295A
- Authority
- CN
- China
- Prior art keywords
- polymer
- carrier
- poly
- preparation
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229920000642 polymer Polymers 0.000 title claims abstract description 101
- 239000000693 micelle Substances 0.000 title claims abstract description 61
- 238000002360 preparation method Methods 0.000 title claims abstract description 46
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 38
- 201000011510 cancer Diseases 0.000 title claims abstract description 23
- 230000004044 response Effects 0.000 title claims description 19
- 230000008685 targeting Effects 0.000 claims abstract description 40
- 239000003446 ligand Substances 0.000 claims abstract description 33
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 32
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 claims abstract description 28
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000003814 drug Substances 0.000 claims abstract description 24
- 229940079593 drug Drugs 0.000 claims abstract description 13
- 210000000170 cell membrane Anatomy 0.000 claims abstract description 9
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 8
- 239000000090 biomarker Substances 0.000 claims abstract description 6
- 108090000672 Annexin A5 Proteins 0.000 claims abstract description 3
- 102000004121 Annexin A5 Human genes 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 59
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 27
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 25
- 229920001223 polyethylene glycol Polymers 0.000 claims description 20
- 239000002202 Polyethylene glycol Substances 0.000 claims description 19
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 19
- 229960001760 dimethyl sulfoxide Drugs 0.000 claims description 14
- -1 polyoxyethylene Polymers 0.000 claims description 13
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- 239000004793 Polystyrene Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 9
- 229920001244 Poly(D,L-lactide) Polymers 0.000 claims description 8
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 8
- 239000013049 sediment Substances 0.000 claims description 8
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 7
- 239000007853 buffer solution Substances 0.000 claims description 7
- 229960002885 histidine Drugs 0.000 claims description 7
- 230000004048 modification Effects 0.000 claims description 7
- 238000012986 modification Methods 0.000 claims description 7
- 238000006116 polymerization reaction Methods 0.000 claims description 7
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 6
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 229910021538 borax Inorganic materials 0.000 claims description 6
- 150000002170 ethers Chemical class 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N hydrochloric acid Substances Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 235000010339 sodium tetraborate Nutrition 0.000 claims description 6
- 238000000967 suction filtration Methods 0.000 claims description 6
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 claims description 6
- BSVBQGMMJUBVOD-UHFFFAOYSA-N trisodium borate Chemical compound [Na+].[Na+].[Na+].[O-]B([O-])[O-] BSVBQGMMJUBVOD-UHFFFAOYSA-N 0.000 claims description 6
- 150000002148 esters Chemical class 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 4
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 4
- 239000007795 chemical reaction product Substances 0.000 claims description 4
- 238000004090 dissolution Methods 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- AYMLQYFMYHISQO-QMMMGPOBSA-N (2s)-3-(1h-imidazol-3-ium-5-yl)-2-[(2-methylpropan-2-yl)oxycarbonylamino]propanoate Chemical compound CC(C)(C)OC(=O)N[C@H](C(O)=O)CC1=CN=CN1 AYMLQYFMYHISQO-QMMMGPOBSA-N 0.000 claims description 3
- 125000001917 2,4-dinitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C(=C1*)[N+]([O-])=O)[N+]([O-])=O 0.000 claims description 3
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 3
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 3
- 150000001299 aldehydes Chemical class 0.000 claims description 3
- 150000001412 amines Chemical class 0.000 claims description 3
- 239000011258 core-shell material Substances 0.000 claims description 3
- 238000000502 dialysis Methods 0.000 claims description 3
- DHQUQYYPAWHGAR-UHFFFAOYSA-N dibenzyl 2-aminopentanedioate Chemical compound C=1C=CC=CC=1COC(=O)C(N)CCC(=O)OCC1=CC=CC=C1 DHQUQYYPAWHGAR-UHFFFAOYSA-N 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 229920001427 mPEG Polymers 0.000 claims description 3
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 3
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 3
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 2
- 125000002252 acyl group Chemical group 0.000 claims description 2
- 229910021529 ammonia Inorganic materials 0.000 claims description 2
- 239000003550 marker Substances 0.000 claims description 2
- 229910052698 phosphorus Inorganic materials 0.000 claims description 2
- 239000011574 phosphorus Substances 0.000 claims description 2
- 229920001610 polycaprolactone Polymers 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims 2
- XDQGMXYCBZNEAG-UHFFFAOYSA-N C(C)[C]CCCN(C)C Chemical compound C(C)[C]CCCN(C)C XDQGMXYCBZNEAG-UHFFFAOYSA-N 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 claims 1
- 239000006071 cream Substances 0.000 claims 1
- 230000006837 decompression Effects 0.000 claims 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N isopropyl alcohol Natural products CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 238000006386 neutralization reaction Methods 0.000 claims 1
- 239000000376 reactant Substances 0.000 claims 1
- 150000003839 salts Chemical class 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 5
- 239000002246 antineoplastic agent Substances 0.000 abstract description 3
- 229940041181 antineoplastic drug Drugs 0.000 abstract description 3
- 230000002708 enhancing effect Effects 0.000 abstract description 2
- 230000008595 infiltration Effects 0.000 abstract description 2
- 238000001764 infiltration Methods 0.000 abstract description 2
- 238000011068 loading method Methods 0.000 abstract description 2
- 230000014759 maintenance of location Effects 0.000 abstract description 2
- 230000006872 improvement Effects 0.000 description 9
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 6
- 239000001257 hydrogen Substances 0.000 description 6
- 101100353912 Arabidopsis thaliana PSBP1 gene Proteins 0.000 description 5
- 101100245627 Nicotiana tabacum PSBP2 gene Proteins 0.000 description 5
- 101150035845 PSBP gene Proteins 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 239000012528 membrane Substances 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- OVBPIULPVIDEAO-LBPRGKRZSA-N Folic acid Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 3
- 229920000383 Poly(ethylene glycol) methyl ether-block-poly(D,L lactide) Polymers 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 229920001577 copolymer Polymers 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 229930012538 Paclitaxel Natural products 0.000 description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000019152 folic acid Nutrition 0.000 description 2
- 239000011724 folic acid Substances 0.000 description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229960001592 paclitaxel Drugs 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 229920000747 poly(lactic acid) Polymers 0.000 description 2
- 239000004626 polylactic acid Substances 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- 150000003462 sulfoxides Chemical class 0.000 description 2
- 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 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- PXFBZOLANLWPMH-UHFFFAOYSA-N 16-Epiaffinine Natural products C1C(C2=CC=CC=C2N2)=C2C(=O)CC2C(=CC)CN(C)C1C2CO PXFBZOLANLWPMH-UHFFFAOYSA-N 0.000 description 1
- KGIGUEBEKRSTEW-UHFFFAOYSA-N 2-vinylpyridine Chemical compound C=CC1=CC=CC=N1 KGIGUEBEKRSTEW-UHFFFAOYSA-N 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102000001189 Cyclic Peptides Human genes 0.000 description 1
- 108010069514 Cyclic Peptides Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 235000016408 Podocarpus macrophyllus Nutrition 0.000 description 1
- 244000162450 Taxus cuspidata Species 0.000 description 1
- 235000009065 Taxus cuspidata Nutrition 0.000 description 1
- 102000009618 Transforming Growth Factors Human genes 0.000 description 1
- 108010009583 Transforming Growth Factors Proteins 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 229940064302 folacin Drugs 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 125000003827 glycol group Chemical group 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 201000005296 lung carcinoma Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 210000000865 mononuclear phagocyte system Anatomy 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 125000001095 phosphatidyl group Chemical group 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000010695 polyglycol Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000010837 receptor-mediated endocytosis Effects 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 230000010148 water-pollination Effects 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses the polymer micelles and preparation method thereof that a kind of cancer target and pH are responded, including having the ligand modified first polymer carrier of targeting ability to kinds of tumor cells film surface biomarker phosphatidyl serine, the ligand is Annexin V, PSBP-6, hexapeptide, one of PGN635, the first polymer carrier is hydrophilic section, the amphipathic nature block polymer that hydrophobic section is formed, preparation method includes the preparation process for the first polymer carrier modified the targeting ligand that cell membrane surface biomarker phosphatidyl serine has affinity；Polymer micelle is actively accumulated in tumor tissues by enhancing infiltration retention effect, longer second polymer carrier depolymerizes in mixed micelle structure, expose targeting ligand, selectively targeted tumour cell, as antineoplastic drug carrier, there is high stability, high Drug loading capacity, long-acting slow-releasing and controlled-releasing ability, high targeting ability.
Description
Technical field
The invention belongs to tumour medicine targeting delivery vehicles and preparation method thereof technical fields, and in particular to be a kind of swollen
The polymer micelle and preparation method thereof of tumor targeting and pH response.
Background technique
Compared with normal cell, tumour cell is overexpressed some receptors, including folacin receptor, transforming growth factor receptor and
EGF-R ELISA realizes carrier active targeting energy through carrier surface ligands specific in conjunction with tumor surface receptor
Power.In carrier surface modified specificity ligand, drug tumour cell accumulating capability is improved, is increased by receptor mediated endocytosis
Add intake of the tumour cell to carrier, improving drug effect reduces toxic side effect.
Amphipathilic block polymer micelle refers to by the self assembly in aqueous solution of the block polymer with amphipathic structure
One kind of formation has core-shell structure, and using hydrophobic group as kernel, hydrophilic group is the molecule aggregate of shell.Its hydrophobic cores can
Insoluble drug is loaded as drug release reservoir, hydrophily shell can be reduced between micella and mononuclear phagocyte system and be interacted,
It avoids being absorbed by mononuclear macrophage, extends circulation time in vivo.Use Amphipathilic block polymer micelle anti-swollen as slightly solubility
Tumor medicine carrier has high stability, high Drug loading capacity, long-acting slow-releasing and controlled-releasing ability, high targeting ability.Become polymer micelle
One of the main approaches of insoluble anti-tumor medicament drug delivery system.
Phosphatidylserine (pHospHatidylserine, PS) belongs to the glycerophosphatide class in cytoplasma membrane phosphatide, is
Important one of the phospholipid composition of cytoplasma membrane, under normal physiological conditions, PS, which is generally concentrated, is distributed in cytoplasma membrane endite, but
It is scientific research personnel's discovery in recent years, PS, which turns up, to be present on the outside of kinds of tumor cells, such as lung cancer, breast cancer, bladder cancer, pancreas
Cancer, cutaneum carcinoma, metastatic encephaloma, carcinoma of the rectum etc., and the amount of its surface expression PS and the grade malignancy of tumour cell are positively correlated
Property.It has been reported that PS is efficiently used for the diagnosis and tumour cell targeting of cancer as tumour cell biomarker,
For targeted pancreatic cancer lung carcinoma cell etc..Research finds that the polypeptide FNFRLKAGQKIRFG (PSBP-0) containing 14 amino acid can be with
PS that cell membrane surface turns up is combined, but affinity is extremely low, after Gln in PSBP-0 amino acid sequence substituted with Ala obtain
The affinity of tumor cell surface PS is remarkably reinforced in PSBP-6.
Summary of the invention
Technical problem to be solved by the present invention lies in: the polymer micelle of a kind of cancer target and pH response, energy are provided
Enough selectively targeted tumour cells, improving anti-tumor drug drug effect reduces toxic side effect.
The present invention is to solve above-mentioned technical problem by the following technical programs:
A kind of polymer micelle of cancer target and pH response, including to kinds of tumor cells film surface biomarker phosphorus
Ester acyl serine have targeting ability ligand modified first polymer carrier, the ligand be Annexin V, PSBP-6,
One of hexapeptide, PGN635, the first polymer carrier are hydrophilic section, the amphipathic block that hydrophobic section is formed
Copolymer.
As an improvement of the above technical solution, the hydrophilic section is polyethylene glycol, in polyoxyethylene, polyvinylpyrrolidone
Any substance, the hydrophobic section be poly-dl-lactide, polystyrene, poly-epsilon-caprolactone, poly- benzyl asparatate,
Any substance in poly- benzyl glutamate, poly lactide-glycolide acid.
As an improvement of the above technical solution, the polymer micelle further includes being collectively formed to mix with the first vector
The second polymer carrier of micella；
Wherein, histidine protonates under the slightly sour environment of tumour in the second polymer carrier, structural instability, occurs
It depolymerizes.
As an improvement of the above technical solution, the second polymer carrier includes using polyhistidyl and polyethylene glycol group
At amphipathic nature block polymer.
As an improvement of the above technical solution, the second polymer carrier further includes the glue of pH sensitive chemicals key connection
Beam, the pH sensitive chemicals key are one of hydrazone bond, second dicarboxyl aldehyde key, β-carboxylic acyloxy amine key, original acid ester key.
Meanwhile the preparation method of the above-mentioned cancer target of this hair disclosure of the invention and the polymer micelle of pH response, preparation side
Method includes the first polymer modified the targeting ligand that cell membrane surface biomarker phosphatidyl serine has affinity
The preparation method of carrier；
Wherein, when in the ligand molecular structure that the phosphatidyl serine to turn up to tumour cell film surface has targeting ability
When containing active amino, preparation method includes the following steps:
1.1, it is activated using 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide
Terminal carboxyl group in the first polymer carrier；
1.2, there is the carboxyl activated in step 1.1 with the phosphatidyl serine to turn up to tumour cell film surface affine
Amino is crosslinked in the targeting ligand of power；
1.3, it dialyses to the reaction product in step 1.2, acquired solution is freeze-dried up to outside to cell membrane surface
The phosphatidyl serine turned over has the carrier of targeting affinity；
Wherein, when in the ligand molecular structure that the phosphatidyl serine to turn up to tumour cell film surface has targeting ability
When without containing active amino, preparation method includes the following steps:
2.1, it is activated using 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide
There is the terminal carboxyl group of the ligand of targeting ability to the phosphatidyl serine that tumour cell film surface is turned up；
2.2, it is condensed the amino in the carboxyl activated in step 2.1 and the first polymer carrier；
2.3, it dialyses to the reaction product of step 2.2, acquired solution is freeze-dried to turn up to cell membrane surface
Phosphatidyl serine have targeting affinity carrier.
As an improvement of the above technical solution, the preparation process including PSBP-6 modification first polymer carrier, and it is described
First polymer carrier is the amphipathic nature block polymer that poly-dl-lactide and polyethylene glycol are formed, preparation process such as I institute of formula
Show:
First polymer carrier is the preparation side for the amphipathic nature block polymer that poly-dl-lactide and polyethylene glycol are formed
Method, comprising the following steps:
PDLLA-PEG-COOH is dissolved in dimethyl sulfoxide by step 1.1, is added 1- ethyl-(3- dimethylaminopropyl)
Carbodiimide hydrochloride reacts 5 hours, adds n-hydroxysuccinimide and is stirred overnight at room temperature, obtains PDLLA-PEG-CONHS；
PSBP-6 is dissolved in dimethyl sulphoxide solution and is added dropwise to step 1.1 solution by step 1.2, adds triethylamine tune
PH to 8 or so is saved, is stirred at room temperature 3-5 days；
Step 1.3 dialyses step 1.2 acquired solution, and the racemic of PSBP-6 modification is obtained after acquired solution freeze-drying
The amphipathic nature block polymer that polylactic acid and polyethylene glycol are formed.
It as an improvement of the above technical solution, further include being protonated under the slightly sour environment of tumour, structural instability solves
Polymerization, and the preparation process of the second polymer carrier of mixed micelle can be formed with the first polymer carrier, described the
Dimerization object carrier is the amphipathic nature block polymer that polyhistidyl and polyethylene glycol are formed, and preparation process is as shown in formula II:
The second polymer carrier is the preparation packet for the amphipathic nature block polymer that polyhistidyl and polyethylene glycol are formed
Include following steps:
Step 2.1, N- (tertbutyloxycarbonyl) -1- (2,4- bis- that Boc-His (Dnp)-OHIPAl is dissolved in isopropanol
Nitrobenzophenone)-L-Histidine is dissolved in Isosorbide-5-Nitrae-dioxane, add solution after thionyl chloride Isosorbide-5-Nitrae-dioxane dilution
After being reacted 1.5 hours under nitrogen protection, enough anhydrous ethers are added and sufficiently precipitate, is dried in vacuo sediment 3-4 hours after suction filtration,
Product is dissolved in ether and is recrystallized with 100 DEG C of acetone, after acquired solution is filtered to remove insoluble matter, adds enough anhydrous ethers sufficiently heavy
It forms sediment, is dried in vacuo sediment 3-4 hours after suction filtration, obtains Dnp-NCAHcl；
Sodium carbonate is added after step 2.1 gained Dnp-NCAHcl to be dissolved in N,N-dimethylformamide dissolution in step 2.2
Hydrochloric acid is neutralized, after being stirred at room temperature 1 hour, methoxy poly (ethylene glycol) amine (mPEG-NH is added2) N,N-dimethylformamide solution
It is stirred to react under reduced pressure and is centrifuged and is filtered with filter paper after a week, solution is added enough ether and sufficiently precipitates, after standing overnight
Solution is poured out, bottom precipitation is dried in vacuo 1-2 days to obtain brown solid PEG-b-poly (Nim-Dnp-His)；
Step 2.3, by PEG-b-poly made from step 2.2 (Nim-Dnp-His) be added N,N-dimethylformamide it is molten
Xie Hou is added beta -mercaptoethanol and reacts 2-3 days, sloughs 2,4- dinitrophenyl and acquired solution is fitted into bag filter in dimethyl
It dialyses 6 hours in sulfoxide, transferring to dialysis in pure water, to dialyzate substantially without yellow, acquired solution is to obtain the final product poly- after being freeze-dried
Histidine and polyethylene glycol amphipathic nature block polymer.
It as an improvement of the above technical solution, further include that the phosphatidyl serine to turn up to tumour cell film surface has target
The preparation process of mixed micelle is formed to the ligand modified first polymer carrier and second polymer carrier of ability comprising
Following steps:
The phosphatidyl serine to turn up to tumour cell film surface is had to the first ligand modified polymerization of targeting ability
Object carrier and second polymer carrier are dissolved in dimethyl sulfoxide, and the sodium borate buffer solution that pH is 9 is added dropwise, is vigorously stirred 1
Hour, bag filter is transferred to after ice-bath ultrasonic and is dialysed with dobell's solution, and mixed micelle solution is obtained.
As an improvement of the above technical solution, further include method that core-shell structure mixed micelle contains drug:
The phosphatidyl serine to turn up to tumour cell film surface is had to the first ligand modified polymerization of targeting ability
Object carrier and second polymer carrier and drug are dissolved in dimethyl sulfoxide, and the sodium borate buffer solution that pH is 9 is added dropwise, acute
Strong stirring 1 hour, is transferred to bag filter and is dialysed with dobell's solution, must carry medicine mixed micelle solution after ice-bath ultrasonic.
The present invention has the advantage that compared with prior art
The present invention discloses a kind of cancer target and pH responsive polymer micella, is based on kinds of tumor cells film surface phosphatidyl
In the expression of serine height and the slightly sour environmental basis of tumor tissues, using the target with phosphatidyl serine with specific affinity
To ligand modified first polymer carrier, mixed micelle is collectively formed with the second polymer carrier of the response slightly sour environment of tumour.
Tumor vessel special construction make injection drug containing carrier solution by enhancing infiltration retention effect actively accumulation in
Tumor tissues, longer second polymer carrier depolymerizes in mixed micelle structure, exposes second polymer carrier end and repairs
The targeting ligand of decorations, selectively targeted tumour cell.
Meanwhile the invention discloses the preparation methods of above-mentioned tumour target polymer micella, including raw to cell membrane surface
The preparation process for the first polymer carrier that there is object marker phosphatidyl serine the targeting ligand of affinity to modify.
Tumour target polymer micella disclosed by the invention has high stability, high load medicine as antineoplastic drug carrier
Ability, long-acting slow-releasing and controlled-releasing ability, high targeting ability.
Detailed description of the invention
Fig. 1 is the hydrogen spectrogram of the PDLLA-PEG-COOH in the embodiment of the present invention；
Fig. 2 is the hydrogen spectrogram of the PDLLA-PEG-PSBP-6 in the embodiment of the present invention；
Fig. 3 is the hydrogen spectrogram of the PEG-pHis in the embodiment of the present invention；
Fig. 4 is the blank mixed micelle partial size of the PDLLA-PEG-PSBP-6 and PEG-pHis formation in the embodiment of the present invention
Distribution schematic diagram；
Fig. 5 is the load medicine mixed micelle partial size of the PDLLA-PEG-PSBP-6 and PEG-pHis formation in the embodiment of the present invention
Distribution schematic diagram；
Fig. 6 is the mixed micelle flow cytometric analysis results figure in the embodiment of the present invention；
Wherein, Fig. 6 includes the mixed micelle under the conditions of five kinds of different proportions of a1-a5, specifically:
A1:PSBP:PDLLA-PEG-COOH=1:1；
A2:PSBP:PDLLA-PEG-COOH=1:3；
A3:PSBP:PDLLA-PEG-COOH=1:5；
A4:PSBP:PDLLA-PEG-COOH=1:7；
A5: PSBP is not contained；
Fig. 7 is that the mixed micelle of the different graft ratios in the embodiment of the present invention absorbs the percentage result table of comparisons.
Specific embodiment
It elaborates below to the embodiment of the present invention, the present embodiment carries out under the premise of the technical scheme of the present invention
Implement, the detailed implementation method and specific operation process are given, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1: the preparation of cancer target and the polymer micelle of pH response:
Poly-dl-lactide that PDLLA-PEG-PSBP-6 described below, which is PSBP-6, to be modified as targeting ligand and poly-
Glycol copolymer, the pHis are polyhistidyl and ethylene glycol copolymer.
The preparation method of a kind of cancer target and the polymer micelle of pH response, the system including PDLLA-PEG-PSBP-6
Standby, PEG-pHis preparation, and PDLLA-PEG-PSBP-6 and PEG-pHis are prepared into mixed micelle.
The preparation process of above-mentioned PDLLA-PEG-PSBP-6 is as shown in formula I:
The preparation of above-mentioned PDLLA-PEG-PSBP-6 the following steps are included:
PDLLA-PEG-COOH is dissolved in dimethyl sulfoxide by step 1.1, is added 1- ethyl-(3- dimethylaminopropyl)
Carbodiimide hydrochloride reacts 5 hours, adds n-hydroxysuccinimide and is stirred overnight at room temperature, obtains PDLLA-PEG-CONHS；
PSBP-6 is dissolved in dimethyl sulphoxide solution and is added dropwise to step 1.1 solution by step 1.2, adds triethylamine tune
PH to 8 or so is saved, is stirred at room temperature 3-5 days；
Step 1.3 dialyses step 1.2 acquired solution, and the racemic of PSBP-6 modification is obtained after acquired solution freeze-drying
The amphipathic nature block polymer that polylactic acid and polyethylene glycol are formed.
The preparation process of PEG-pHis is as shown in formula II:
The preparation method of PEG-pHis the following steps are included:
Step 2.1, N- (tertbutyloxycarbonyl) -1- (2,4- bis- that Boc-His (Dnp)-OHIPAl is dissolved in isopropanol
Nitrobenzophenone)-L-Histidine is dissolved in Isosorbide-5-Nitrae-dioxane, add solution after thionyl chloride Isosorbide-5-Nitrae-dioxane dilution
After being reacted 1.5 hours under nitrogen protection, enough anhydrous ethers are added and sufficiently precipitate, is dried in vacuo sediment 3-4 hours after suction filtration,
Product is dissolved in ether and is recrystallized with 100 DEG C of acetone, after acquired solution is filtered to remove insoluble matter, adds enough anhydrous ethers sufficiently heavy
It forms sediment, is dried in vacuo sediment 3-4 hours after suction filtration, obtains Dnp-NCAHcl；
Sodium carbonate is added after step 2.1 gained Dnp-NCAHcl to be dissolved in N,N-dimethylformamide dissolution in step 2.2
Hydrochloric acid is neutralized, after being stirred at room temperature 1 hour, methoxy poly (ethylene glycol) amine (mPEG-NH is added2) N,N-dimethylformamide solution
It is stirred to react under reduced pressure and is centrifuged and is filtered with filter paper after a week, solution is added enough ether and sufficiently precipitates, after standing overnight
Solution is poured out, bottom precipitation is dried in vacuo 1-2 days to obtain brown solid PEG-b-poly (Nim-Dnp-His)；
Step 2.3, by PEG-b-poly made from step 2.2 (Nim-Dnp-His) be added N,N-dimethylformamide it is molten
Xie Hou is added beta -mercaptoethanol and reacts 2-3 days, sloughs 2,4- dinitrophenyl and acquired solution is fitted into bag filter in dimethyl
It dialyses 6 hours in sulfoxide, transferring to dialysis in pure water, to dialyzate substantially without yellow, acquired solution is to obtain the final product poly- after being freeze-dried
Histidine and polyethylene glycol amphipathic nature block polymer.
The hydrogen spectrum of PDLLA-PEG-COOH is as shown in Figure 1；
The hydrogen spectrum of PDLLA-PEG-PSBP-6 is as shown in Figure 2；
The hydrogen spectrum of PEG-pHis is as shown in Figure 3.
PDLLA-PEG-PSBP-6 and PEG-pHis are prepared into mixed micelle, the preparation method of mixed micelle includes following
Step:
The phosphatidyl serine to turn up to tumour cell film surface is had to the first ligand modified polymerization of targeting ability
Object carrier and second polymer carrier are dissolved in dimethyl sulfoxide, and the sodium borate buffer solution that pH is 9 is added dropwise, is vigorously stirred 1
Hour, bag filter is transferred to after ice-bath ultrasonic and is dialysed with dobell's solution, and mixed micelle solution is obtained.
The blank mixed micelle particle diameter distribution that PDLLA-PEG-PSBP-6 and PEG-pHis is formed is shown as shown in Figure 4.
Embodiment 2: the polymer micelle that cancer target and pH are responded contains the preparation of drug
The phosphatidyl serine to turn up to tumour cell film surface is had to the first ligand modified polymerization of targeting ability
Object carrier and second polymer carrier and drug are dissolved in dimethyl sulfoxide, and the sodium borate buffer solution that pH is 9 is added dropwise, acute
Strong stirring 1 hour, is transferred to bag filter and is dialysed with dobell's solution, must carry medicine mixed micelle solution after ice-bath ultrasonic.
Specifically, containing Japanese yew as carrier medicament, cancer target and the pH polymer micelle responded using taxol
The method that the preparation of alcohol contains drug is as follows:
PDLLA-PEG-PSBP-6, PEG-pHis and taxol are dissolved in dimethyl sulfoxide, the boric acid that pH is 9 is added dropwise
Sodium buffer solution is vigorously stirred 1 hour, and ice-bath ultrasonic is transferred to bag filter dobell's solution and dialyses 24 hours after 30 minutes leads to
The filtering of 0.45 micron membrane filter is crossed to get medicine mixed micelle solution is carried.
The load medicine mixed micelle particle diameter distribution that PDLLA-PEG-PSBP-6 and PEG-pHis is formed is as shown in Figure 5.
Embodiment 3:
The targeting ability of above-mentioned PDLLA-PEG-PSBP-6, PEG-pHis mixed micelle being prepared, uses with lower section
Method verifying:
PDLLA-PEG-PSBP-6, PEG-pHis mixed micelle target ability and investigate:
HELA cell, which uses, contains 10% fetal calf serum DMEM culture medium, and 37 DEG C of 5%CO2 are cultivated in incubator, take in pair
The cell in number growth period is tested, and is separately added into PDLLA-PEG-FITC, PDLLA-PEG-PSBP-6 and PEG-pHis and is formed
Mixed micelle and PDLLA-PEG-FITC, PDLLA-PEG-COOH and PEG-pHis formed mixed micelle, act on 4 hours
Afterwards, phosphate buffer washes away the fluorescent material not absorbed, analyzes two groups with stream type cell analyzer after 4% paraformaldehyde is fixed
Fluorescence intensity is absorbed, every group sets three multiple holes, as a result as shown in Figure 6.
As a result the PDLLA-PEG-PSBP-6 and PEG- being grafted with molar ratio PSBP-6:COOH-PEG-PDLLA=1:1
It is 83.7 ± 2 that pHis mixed micelle group, which absorbs percentage, and intake average fluorescent strength is 5729 ± 351.7；The intake of 1:3 grafting
Percentage is 89.3 ± 1.4, and intake average fluorescent strength is 6969 ± 170；The intake percentage of 1:5 grafting is 60.0 ± 3.2,
Absorbing average fluorescent strength is 4915 ± 252.6；The intake percentage of 1:7 grafting is 51.5 ± 0.8, absorbs average fluorescent strength
It is 4499.6 ± 50.6；It is 25.5 ± 1.76 that PDLLA-PEG-COOH and PEG-pHis mixed micelle group, which absorbs percentage, intake
Average fluorescent strength is 3263.7 ± 129.7, and the mixed micelle intake percentage result table of comparisons of above-mentioned difference graft ratio is such as
Shown in Fig. 7.
The result shows that:
Firstly, from Fig. 4-5: PEG-pHis has the targeting conveying capacity for tumor cell tissue；
Secondly, as shown in Figure 6: after PSBP-6 modification PEG-PDLLA, than unmodified PEG- under the same terms
PDLLA intake dramatically increases,
Finally, as shown in Figure 7: having with PSBP-6:COOH-PEG-PDLLA 1:3 grafting PDLLA-PEG-PSBP6 optimal
Targeting Effect, this is consistent with the characteristics of PSBP-6 set forth in the present invention.
In the present invention, have the ligand of targeting ability can also be in folic acid, PSBP-6, RGD cyclic peptide tumour cell
It is a kind of.
In the present invention, first polymer carrier PDLLA-PEG is also possible to the carrier material with hydrophilic section and hydrophobic section,
Hydrophilic section can for polyoxyethylene, polyvinylpyrrolidone etc., hydrophobic section can for l-lactic acid, polystyrene, poly- ε-oneself
Lactone, poly- benzyl asparatate, poly- benzyl glutamate, poly lactide-glycolide acid etc..
It can be poly group ammonia with the PDLLA-PEG-PSBP-6 pH sensitive material that mixed micelle is collectively formed in the present invention
Amphipathic nature block polymer, poly- N, N- diethyl amido ethylmethyl acrylate and the polyethylene glycol of acid and polyethylene glycol composition
The amphipathic nature block polymer of composition, poly- (4- or 2- vinylpyridine) and polyethylene glycol composition amphipathic nature block polymer,
Or the micella of pH sensitive chemicals key connection, including hydrazone bond, second dicarboxyl aldehyde key, β-carboxylic acyloxy amine key, original acid ester key.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (10)
1. the polymer micelle of a kind of cancer target and pH response, which is characterized in that including raw to kinds of tumor cells film surface
Object marker phosphatidyl serine has the ligand modified first polymer carrier of targeting ability, and the ligand is Annexin
V, one of PSBP-6, hexapeptide, PGN635, the first polymer carrier are hydrophilic section, what hydrophobic section was formed
Amphipathic nature block polymer.
2. the polymer micelle of cancer target according to claim 1 and pH response, which is characterized in that the hydrophilic section is
Polyethylene glycol, polyoxyethylene, any substance in polyvinylpyrrolidone, the hydrophobic section is poly-dl-lactide, poly-
Styrene, poly-epsilon-caprolactone, poly- benzyl asparatate, poly- benzyl glutamate, any in poly lactide-glycolide acid
A kind of substance.
3. the polymer micelle of cancer target according to claim 1 to 2 and pH response, which is characterized in that described poly-
Closing object micella further includes the second polymer carrier that mixed micelle is collectively formed with the first vector；
Wherein, histidine protonates under the slightly sour environment of tumour in the second polymer carrier, structural instability, and depolymerization occurs
It closes.
4. the polymer micelle of cancer target according to claim 3 and pH response, which is characterized in that second polymerization
Object carrier includes the amphipathic nature block polymer of polyhistidyl and polyethylene glycol composition.
5. the polymer micelle of cancer target according to claim 4 and pH response, which is characterized in that second polymerization
Object carrier further includes the micella of pH sensitive chemicals key connection, and the pH sensitive chemicals key is hydrazone bond, second dicarboxyl aldehyde key, β-carboxylic
One of sour amido bond, original acid ester key.
6. the preparation method of a kind of cancer target a method as claimed in any one of claims 1 to 5 and the polymer micelle of pH response, special
Sign is that first including having the targeting ligand of affinity to modify to cell membrane surface biomarker phosphatidyl serine gathers
Close the preparation method of object carrier；
Wherein, contain when in the ligand molecular structure that the phosphatidyl serine to turn up to tumour cell film surface has targeting ability
When active amino, preparation method includes the following steps:
1.1, using described in 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide activation
Terminal carboxyl group in first polymer carrier；
1.2, make the carboxyl activated in step 1.1 that there is affinity with the phosphatidyl serine to turn up to tumour cell film surface
Amino is crosslinked in targeting ligand；
1.3, it dialyses to the reaction product in step 1.2, acquired solution is freeze-dried to turn up to cell membrane surface
Phosphatidyl serine has the carrier of targeting affinity；
Wherein, it is free of when in the ligand molecular structure that the phosphatidyl serine to turn up to tumour cell film surface has targeting ability
When active amino, preparation method includes the following steps:
2.1, using 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide activation to swollen
The phosphatidyl serine that oncocyte film surface is turned up has the terminal carboxyl group of the ligand of targeting ability；
2.2, it is condensed the amino in the carboxyl activated in step 2.1 and the first polymer carrier；
2.3, it dialyses to the reaction product of step 2.2, acquired solution is freeze-dried up to the phosphorus to turn up to cell membrane surface
Ester acyl serine has the carrier of targeting affinity.
7. the preparation method of cancer target according to claim 6 and the polymer micelle of pH response, which is characterized in that packet
The preparation process of PSBP-6 modification first polymer carrier is included, and the first polymer carrier is poly-dl-lactide and poly- second
The amphipathic nature block polymer that glycol is formed, preparation process is as shown in formula I:
First polymer carrier is the preparation method packet for the amphipathic nature block polymer that poly-dl-lactide and polyethylene glycol are formed
Include following steps:
PDLLA-PEG-COOH is dissolved in dimethyl sulfoxide by step 1.1, and 1- ethyl-(3- dimethylaminopropyl) carbon two is added
It imide hydrochloride reactant salt 5 hours, adds n-hydroxysuccinimide and is stirred overnight at room temperature, obtain PDLLA-PEG-CONHS；
PSBP-6 is dissolved in dimethyl sulphoxide solution and is added dropwise to step 1.1 solution by step 1.2, is added triethylamine and is adjusted pH
To 8 or so, it is stirred at room temperature 3-5 days；
Step 1.3 dialyses step 1.2 acquired solution, and the poly- cream of racemic of PSBP-6 modification is obtained after acquired solution freeze-drying
The amphipathic nature block polymer that acid is formed with polyethylene glycol.
8. the preparation method of cancer target according to claim 6 or 7 and the polymer micelle of pH response, feature exist
In further including being protonated under the slightly sour environment of tumour, structural instability depolymerizes, and can carry with the first polymer
Body forms the preparation process of the second polymer carrier of mixed micelle, and the second polymer carrier is polyhistidyl and poly- second two
The amphipathic nature block polymer that alcohol is formed, preparation process is as shown in formula II:
The second polymer carrier is the preparation method for the amphipathic nature block polymer that polyhistidyl and polyethylene glycol are formed,
The following steps are included:
Step 2.1, N- (tertbutyloxycarbonyl) -1- (2,4- dinitro that Boc-His (Dnp)-OHIPAl is dissolved in isopropanol
Phenyl)-L-Histidine is dissolved in Isosorbide-5-Nitrae-dioxane, add solution nitrogen after thionyl chloride Isosorbide-5-Nitrae-dioxane dilution
After lower reaction 1.5 hours of protection, enough anhydrous ethers, which are added, sufficiently to be precipitated, and is dried in vacuo sediment 3-4 hours after suction filtration, product
It is dissolved in ether and is recrystallized with 100 DEG C of acetone, after acquired solution is filtered to remove insoluble matter, enough anhydrous ethers is added sufficiently to precipitate,
It is dried in vacuo sediment 3-4 hours after suction filtration, obtains Dnp-NCAHcl；
Sodium carbonate neutralization is added after step 2.1 gained Dnp-NCAHcl to be dissolved in N,N-dimethylformamide dissolution in step 2.2
After being stirred at room temperature 1 hour, methoxy poly (ethylene glycol) amine (mPEG-NH is added in hydrochloric acid2) N,N-dimethylformamide solution decompression
Under the conditions of be stirred to react and be centrifuged and filtered with filter paper after a week, solution is added enough ether and sufficiently precipitates, will be molten after standing overnight
Liquid is poured out, and bottom precipitation is dried in vacuo 1-2 days to obtain brown solid PEG-b-poly (Nim-Dnp-His)；
Step 2.3, by PEG-b-poly made from step 2.2 (Nim-Dnp-His) be added n,N-Dimethylformamide dissolution after,
Beta -mercaptoethanol is added to react 2-3 days, sloughs 2,4- dinitrophenyl and acquired solution is fitted into bag filter in dimethyl sulfoxide
Middle dialysis 6 hours is transferred in pure water and is dialysed to dialyzate substantially without yellow, up to poly group ammonia after acquired solution freeze-drying
Acid and polyethylene glycol amphipathic nature block polymer.
9. the preparation method of cancer target according to claim 8 and the polymer micelle of pH response, which is characterized in that also
Phosphatidyl serine including turning up to tumour cell film surface has the ligand modified first polymer carrier of targeting ability
The preparation process of mixed micelle is formed with second polymer carrier comprising following steps:
There is the ligand modified first polymer of targeting ability to carry the phosphatidyl serine to turn up to tumour cell film surface
Body and second polymer carrier are dissolved in dimethyl sulfoxide, and the sodium borate buffer solution that pH is 9 is added dropwise, it is small to be vigorously stirred 1-2
When, bag filter is transferred to after ice-bath ultrasonic and is dialysed with dobell's solution, and mixed micelle solution is obtained.
10. the preparation method of cancer target according to claim 9 and the polymer micelle of pH response, which is characterized in that
Further include the method that core-shell structure mixed micelle contains drug:
There is the ligand modified first polymer of targeting ability to carry the phosphatidyl serine to turn up to tumour cell film surface
Body and second polymer carrier and drug are dissolved in dimethyl sulfoxide, and the sodium borate buffer solution that pH is 9 is added dropwise, acutely stirs
It mixes 1-2 hours, bag filter is transferred to after ice-bath ultrasonic and is dialysed with dobell's solution, medicine mixed micelle solution must be carried.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811248768.5A CN109833295A (en) | 2018-10-25 | 2018-10-25 | A kind of polymer micelle and preparation method thereof of cancer target and pH response |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811248768.5A CN109833295A (en) | 2018-10-25 | 2018-10-25 | A kind of polymer micelle and preparation method thereof of cancer target and pH response |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109833295A true CN109833295A (en) | 2019-06-04 |
Family
ID=66883074
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811248768.5A Pending CN109833295A (en) | 2018-10-25 | 2018-10-25 | A kind of polymer micelle and preparation method thereof of cancer target and pH response |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109833295A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111848964A (en) * | 2020-06-30 | 2020-10-30 | 无锡市妇幼保健院 | CD44 targeted and pH responsive drug-loaded nano-micelle and preparation method and application thereof |
| CN112107690A (en) * | 2020-10-16 | 2020-12-22 | 浙江工业大学 | Preparation method of camptothecin drug nanoparticles |
| CN112876673A (en) * | 2021-01-25 | 2021-06-01 | 山东大学 | PH-responsive nano copolymer carrier and preparation method and application thereof |
| EP4127208A4 (en) * | 2020-04-03 | 2024-06-05 | University of Cincinnati | Secreted phosphorylated heat shock protein-70 as a biomarker for treating and diagnosing cancer |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101848702A (en) * | 2006-12-01 | 2010-09-29 | 安特里奥公司 | Micellar nanoparticles comprising botulinum toxin |
| CN103370329A (en) * | 2010-12-20 | 2013-10-23 | Cn制药株式会社 | Amphiphilic cyclic phosphazene trimer, hydrophobic pharmaceutical formulation micellized by amphiphilic cyclic phosphazene trimer, and preparation methods thereof |
| WO2013184976A3 (en) * | 2012-06-06 | 2015-06-25 | Northwestern University | Compositions and methods for antigen-specific tolerance |
| CN105168130A (en) * | 2015-08-06 | 2015-12-23 | 安徽医科大学 | Tumor-targeted polymer micelle and preparation method thereof |
-
2018
- 2018-10-25 CN CN201811248768.5A patent/CN109833295A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101848702A (en) * | 2006-12-01 | 2010-09-29 | 安特里奥公司 | Micellar nanoparticles comprising botulinum toxin |
| CN103370329A (en) * | 2010-12-20 | 2013-10-23 | Cn制药株式会社 | Amphiphilic cyclic phosphazene trimer, hydrophobic pharmaceutical formulation micellized by amphiphilic cyclic phosphazene trimer, and preparation methods thereof |
| WO2013184976A3 (en) * | 2012-06-06 | 2015-06-25 | Northwestern University | Compositions and methods for antigen-specific tolerance |
| CN105168130A (en) * | 2015-08-06 | 2015-12-23 | 安徽医科大学 | Tumor-targeted polymer micelle and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| AMANDA PERREAULT: "Targeting Phosphatidylserine with a 64Cu-Labeled Peptide for Molecular Imaging of Apoptosis", 《MOLECULAR PHARMACEUTICS》 * |
| CHIYI XIONG: "Peptide-Based Imaging Agents Targeting Phosphatidylserine for the Detection of Apoptosis", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4127208A4 (en) * | 2020-04-03 | 2024-06-05 | University of Cincinnati | Secreted phosphorylated heat shock protein-70 as a biomarker for treating and diagnosing cancer |
| CN111848964A (en) * | 2020-06-30 | 2020-10-30 | 无锡市妇幼保健院 | CD44 targeted and pH responsive drug-loaded nano-micelle and preparation method and application thereof |
| CN112107690A (en) * | 2020-10-16 | 2020-12-22 | 浙江工业大学 | Preparation method of camptothecin drug nanoparticles |
| CN112876673A (en) * | 2021-01-25 | 2021-06-01 | 山东大学 | PH-responsive nano copolymer carrier and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chandran et al. | Characterization of a targeted nanoparticle functionalized with a urea-based inhibitor of prostate-specific membrane antigen (PSMA) | |
| CN109833295A (en) | A kind of polymer micelle and preparation method thereof of cancer target and pH response | |
| Fan et al. | POSS-based supramolecular amphiphilic zwitterionic complexes for drug delivery | |
| Liu et al. | Inhibition of murine breast cancer growth and metastasis by survivin-targeted siRNA using disulfide cross-linked linear PEI | |
| CN105463024B (en) | Polyethyleneimine-cyclodextrin-based multifunctional supramolecular gene delivery system and preparation method thereof | |
| US10576038B2 (en) | Method for preparation of reducible degradable hyperbranched polymeric micelles | |
| CN103127526A (en) | Tree-like polymer targeting nanometer drug delivery carrier and its preparation method | |
| CN112876578B (en) | Amphiphilic glucan derivative carrier targeting tumor-associated fibroblasts, and preparation and application of pharmaceutical composition of amphiphilic glucan derivative carrier | |
| CN107260706A (en) | Double medicament-carried nano carriers of a kind of cancer target and preparation method thereof | |
| Xiong et al. | Amphiphilic Cholic‐Acid‐Modified Dextran Sulfate and its Application for the Controlled Delivery of Superoxide Dismutase | |
| Yang et al. | pH and redox dual-responsive multifunctional gene delivery with enhanced capability of transporting DNA into the nucleus | |
| CN104146946B (en) | A kind of graphene oxide medicine carrying composite of hepatoma-targeting and preparation method thereof | |
| CN101812178A (en) | Reduction sensitive polyethyleneimine derivative as well as preparation method and application thereof | |
| CN104623685A (en) | Tumor stroma pH sensitive target dendrimer and preparation method thereof | |
| Wang et al. | PEI functionalized cell membrane for tumor targeted and glutathione responsive gene delivery | |
| CN106397765B (en) | The polyethylenimine derivates and its synthetic method of vitamin E modification and application | |
| Tao et al. | Acid/reduction dual-sensitive amphiphilic graft polyurethane with folic acid and detachable poly (ethylene glycol) as anticancer drug delivery carrier | |
| CN106389384B (en) | A kind of preparation method and application of multistage Liver targeting intelligence nanoscale medicine delivery system | |
| CN101081876A (en) | Subcellular organelle target directional Chitosan oligosaccharide-aliphatic acid grafting matter and preparation and application thereof | |
| CN108503845A (en) | Amphiphilic graft polymers and its synthetic method with catechol ligand and application | |
| CN102250348B (en) | Polyethyleneimine derivative and application thereof as gene transfer carrier | |
| CN104162166A (en) | pH sensitive polysaccharide nano drug-loaded micelles and preparation method thereof | |
| CN112057631A (en) | Application of a modified dendrimer in protein intracellular delivery | |
| Wang et al. | Polycation-telodendrimer nanocomplexes for intracellular protein delivery | |
| CN107693505B (en) | A kind of oil-water dual-soluble ROS-sensitive nanoparticles and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190604 |
|
| RJ01 | Rejection of invention patent application after publication |