CN113797336A - 精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和应用 - Google Patents
精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和应用 Download PDFInfo
- Publication number
- CN113797336A CN113797336A CN202111068594.6A CN202111068594A CN113797336A CN 113797336 A CN113797336 A CN 113797336A CN 202111068594 A CN202111068594 A CN 202111068594A CN 113797336 A CN113797336 A CN 113797336A
- Authority
- CN
- China
- Prior art keywords
- photosensitizer
- arginine
- small peptide
- peg
- arg
- 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
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 103
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 62
- 239000003504 photosensitizing agent Substances 0.000 title claims abstract description 57
- 150000001875 compounds Chemical class 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 239000002245 particle Substances 0.000 claims abstract description 28
- 239000003607 modifier Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000008367 deionised water Substances 0.000 claims abstract description 9
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 9
- 239000003960 organic solvent Substances 0.000 claims abstract description 8
- 238000003756 stirring Methods 0.000 claims abstract description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 13
- 125000003118 aryl group Chemical group 0.000 claims description 11
- 230000002378 acidificating effect Effects 0.000 claims description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N dimethyl sulfoxide Natural products CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 8
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims description 7
- 239000004475 Arginine Substances 0.000 claims description 6
- 229940024606 amino acid Drugs 0.000 claims description 5
- 235000001014 amino acid Nutrition 0.000 claims description 5
- 150000001413 amino acids Chemical class 0.000 claims description 5
- 238000001553 co-assembly Methods 0.000 claims description 5
- 150000004032 porphyrins Chemical group 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 4
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 4
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 4
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 4
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 4
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 4
- 229920006022 Poly(L-lactide-co-glycolide)-b-poly(ethylene glycol) Polymers 0.000 claims description 4
- 229920000362 Polyethylene-block-poly(ethylene glycol) Polymers 0.000 claims description 4
- AOBORMOPSGHCAX-UHFFFAOYSA-N Tocophersolan Chemical compound OCCOC(=O)CCC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C AOBORMOPSGHCAX-UHFFFAOYSA-N 0.000 claims description 4
- 238000009825 accumulation Methods 0.000 claims description 4
- 235000004279 alanine Nutrition 0.000 claims description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 4
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- 230000002209 hydrophobic effect Effects 0.000 claims description 4
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 4
- 229960000310 isoleucine Drugs 0.000 claims description 4
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 4
- 125000001624 naphthyl group Chemical group 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 4
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 4
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 claims description 4
- IEQIEDJGQAUEQZ-UHFFFAOYSA-N phthalocyanine Chemical group N1C(N=C2C3=CC=CC=C3C(N=C3C4=CC=CC=C4C(=N4)N3)=N2)=C(C=CC=C2)C2=C1N=C1C2=CC=CC=C2C4=N1 IEQIEDJGQAUEQZ-UHFFFAOYSA-N 0.000 claims description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 4
- 125000005581 pyrene group Chemical group 0.000 claims description 4
- 125000000542 sulfonic acid group Chemical group 0.000 claims description 4
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 4
- 239000002246 antineoplastic agent Substances 0.000 claims description 3
- 229940041181 antineoplastic drug Drugs 0.000 claims description 3
- VFHDWGAEEDVVPD-UHFFFAOYSA-N chembl507897 Chemical group C1=CC(O)=CC=C1C(C1=CC=C(N1)C(C=1C=CC(O)=CC=1)=C1C=CC(=N1)C(C=1C=CC(O)=CC=1)=C1C=CC(N1)=C1C=2C=CC(O)=CC=2)=C2N=C1C=C2 VFHDWGAEEDVVPD-UHFFFAOYSA-N 0.000 claims description 3
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical group [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 claims description 2
- 239000012046 mixed solvent Substances 0.000 claims description 2
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 2
- ZWXXZYMZDVOLND-UHFFFAOYSA-N C12=CC=C(N1)C=C1C=CC(=N1)C=C1C=CC(N1)=CC=1C=CC(N1)=C2.C2(=CC=CC=C2)O Chemical compound C12=CC=C(N1)C=C1C=CC(=N1)C=C1C=CC(N1)=CC=1C=CC(N1)=C2.C2(=CC=CC=C2)O ZWXXZYMZDVOLND-UHFFFAOYSA-N 0.000 claims 1
- 230000000699 topical effect Effects 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 48
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 abstract description 40
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 25
- 239000001301 oxygen Substances 0.000 abstract description 25
- 229910052760 oxygen Inorganic materials 0.000 abstract description 25
- 230000000694 effects Effects 0.000 abstract description 21
- 206010021143 Hypoxia Diseases 0.000 abstract description 18
- 230000007954 hypoxia Effects 0.000 abstract description 12
- 239000003814 drug Substances 0.000 abstract description 9
- 229940079593 drug Drugs 0.000 abstract description 8
- 239000000203 mixture Substances 0.000 abstract description 6
- 230000002829 reductive effect Effects 0.000 abstract description 6
- 238000002428 photodynamic therapy Methods 0.000 abstract description 5
- 238000011068 loading method Methods 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 4
- 230000014759 maintenance of location Effects 0.000 abstract description 3
- 206010067484 Adverse reaction Diseases 0.000 abstract description 2
- 230000006838 adverse reaction Effects 0.000 abstract description 2
- 239000002552 dosage form Substances 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 abstract 1
- 239000003937 drug carrier Substances 0.000 abstract 1
- 239000000825 pharmaceutical preparation Substances 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 52
- 239000000243 solution Substances 0.000 description 50
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 28
- 239000011259 mixed solution Substances 0.000 description 22
- 239000003642 reactive oxygen metabolite Substances 0.000 description 19
- 210000001519 tissue Anatomy 0.000 description 19
- 229920001223 polyethylene glycol Polymers 0.000 description 18
- 239000002202 Polyethylene glycol Substances 0.000 description 14
- 238000011534 incubation Methods 0.000 description 14
- 239000002609 medium Substances 0.000 description 10
- 239000011550 stock solution Substances 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 8
- 230000003834 intracellular effect Effects 0.000 description 8
- 239000006069 physical mixture Substances 0.000 description 8
- 210000004881 tumor cell Anatomy 0.000 description 8
- 230000008859 change Effects 0.000 description 7
- 238000011835 investigation Methods 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 231100000135 cytotoxicity Toxicity 0.000 description 6
- 230000003013 cytotoxicity Effects 0.000 description 6
- 238000009826 distribution Methods 0.000 description 6
- 230000001146 hypoxic effect Effects 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 239000012737 fresh medium Substances 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 108010019160 Pancreatin Proteins 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 238000012377 drug delivery Methods 0.000 description 4
- 238000002296 dynamic light scattering Methods 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 4
- 229940055695 pancreatin Drugs 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 3
- BEVHTVRRVVEMEF-UHFFFAOYSA-N [6'-acetyloxy-4-amino-2',7'-difluoro-5-(methylamino)-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl] acetate Chemical compound C12=CC(F)=C(OC(C)=O)C=C2OC2=CC(OC(C)=O)=C(F)C=C2C21OC(=O)C1=C(N)C(NC)=CC=C21 BEVHTVRRVVEMEF-UHFFFAOYSA-N 0.000 description 3
- 230000035508 accumulation Effects 0.000 description 3
- 229960003121 arginine Drugs 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- 239000006143 cell culture medium Substances 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 230000036284 oxygen consumption Effects 0.000 description 3
- 230000008557 oxygen metabolism Effects 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- XDFNWJDGWJVGGN-UHFFFAOYSA-N 2-(2,7-dichloro-3,6-dihydroxy-9h-xanthen-9-yl)benzoic acid Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC(Cl)=C(O)C=C2OC2=CC(O)=C(Cl)C=C21 XDFNWJDGWJVGGN-UHFFFAOYSA-N 0.000 description 2
- DIJCILWNOLHJCG-UHFFFAOYSA-N 7-amino-2',7'-difluoro-3',6'-dihydroxy-6-(methylamino)spiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound C12=CC(F)=C(O)C=C2OC2=CC(O)=C(F)C=C2C21OC(=O)C1=C(N)C(NC)=CC=C21 DIJCILWNOLHJCG-UHFFFAOYSA-N 0.000 description 2
- 238000011725 BALB/c mouse Methods 0.000 description 2
- 108090000371 Esterases Proteins 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000012964 benzotriazole Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 210000001163 endosome Anatomy 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 125000002795 guanidino group Chemical group C(N)(=N)N* 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 229940124280 l-arginine Drugs 0.000 description 2
- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Chemical compound O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 description 2
- 230000002438 mitochondrial effect Effects 0.000 description 2
- 230000008811 mitochondrial respiratory chain Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000007959 normoxia Effects 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 230000004962 physiological condition Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000004043 responsiveness Effects 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000011269 treatment regimen Methods 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- DVBUCBXGDWWXNY-SFHVURJKSA-N (2s)-5-(diaminomethylideneamino)-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C3=CC=CC=C3C2=C1 DVBUCBXGDWWXNY-SFHVURJKSA-N 0.000 description 1
- 102100035882 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 102000000634 Cytochrome c oxidase subunit IV Human genes 0.000 description 1
- 108090000365 Cytochrome-c oxidases Proteins 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 1
- 102000011779 Nitric Oxide Synthase Type II Human genes 0.000 description 1
- 108010076864 Nitric Oxide Synthase Type II Proteins 0.000 description 1
- 235000005811 Viola adunca Nutrition 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 235000002254 Viola papilionacea Nutrition 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004103 aerobic respiration Effects 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 150000001555 benzenes Chemical group 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000004611 cancer cell death Effects 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- -1 hydrogen ions Chemical class 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000005917 in vivo anti-tumor Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004705 lumbosacral region Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229940126701 oral medication Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- JMANVNJQNLATNU-UHFFFAOYSA-N oxalonitrile Chemical compound N#CC#N JMANVNJQNLATNU-UHFFFAOYSA-N 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 238000001126 phototherapy Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000057 systemic toxicity Toxicity 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000003868 tissue accumulation Effects 0.000 description 1
- 230000008467 tissue growth Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 230000036326 tumor accumulation Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
- A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
- A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nanotechnology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Crystallography & Structural Chemistry (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Medical Informatics (AREA)
- Manufacturing & Machinery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
精氨酸‑小肽复合物与光敏剂共组装纳米粒及其制备方法和应用,属于药物制剂新辅料和新剂型领域,本发明精氨酸‑小肽复合物与光敏剂的质量比为5:1~1:5。将精氨酸‑小肽复合物、光敏剂与PEG修饰剂的混合物溶解到有机溶剂中,在搅拌条件下,将该溶液缓慢滴加到去离子水中,自发形成粒径小而均一的纳米粒。纳米粒可通过渗透与滞留增强效应富集于肿瘤部位;且具有超高的载药量,降低了辅料相关的不良反应和毒性;组成纳米粒的精氨酸‑小肽复合物中的精氨酸残基可有效产生一氧化氮以缓解肿瘤乏氧,进而显著提高光敏剂的活性氧产量及其光动力学治疗效果。
Description
技术领域
本发明属于药物制剂新辅料和新剂型领域,具体涉及精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和其在抗肿瘤药物传递系统中的应用。
背景技术
癌症是当今严重威胁人类生命健康的重大疾病,据世界卫生组织(WHO)统计,全球每年有超过800万人死于癌症。目前,常见的癌症治疗策略有手术切除、化学药物治疗、放射疗法和光疗。与其他疗法相比,光动力学治疗由于具有较小的全身毒性,以及高的治疗选择性而受到患者以及医护人员的青睐。在光动力学治疗过程中,特定波长的光源照射使光敏剂分子发生能量跃迁,随后将能量传递给周围的氧气(O2)形成活性氧(ROS),对细胞造成不可逆的氧化损伤并导致癌细胞死亡。然而,肿瘤组织自身生长速度过快而血管生长却较慢,其血液循环通常无法有效提供足够的氧气供其生长,因此实体瘤内部的氧分压相比于正常组织会明显下降,正常组织的氧分压平均为40mm Hg,低氧组织的氧分压一般小于14mm Hg,有时甚至小于5mm Hg,这严重降低了PDT的肿瘤治疗效率。
目前,研究人员尝试了多种向肿瘤组织“供氧”的策略以缓解其乏氧。例如:1)将过氧化氢酶、二氧化锰和金纳米团簇等物质递送至肿瘤部位,催化过氧化氢(H2O2)分解为O2。然而,大部分的催化剂效率较慢限制了这种方法的应用。2)使用红细胞、血红蛋白和全氟化碳等载体直接将O2运输至肿瘤。这种方法因载氧效率低、O2渗漏以及难以联合给药而受到诟病。3)将氮化碳递送至肿瘤部位将水分解生产O2。然而,这种方法的O2产量较低且生物安全性较差。因此,这些“供氧”策略只能有限缓解肿瘤缺氧,其进一步的应用受到限制。
肿瘤微环境中的氧气浓度不仅取决于外部氧气供应,还取决于自身氧气消耗。因此,可以尝试抑制肿瘤自身的O2消耗缓解其缺氧。肿瘤细胞较为过度活跃的O2代谢主要在线粒体有氧呼吸(Mito-AR)进行。据报道,一氧化氮(NO)可以竞争性的抑制O2与Mito-AR中线粒体复合体Ⅳ(细胞色素c氧化酶,CcO)的结合,进而降低肿瘤细胞中大部分的O2消耗。因此,NO可能是一种潜在的肿瘤缺氧抑制剂。然而,直接运输气体存在装载率低、稳定性差、气体渗漏以及难与其他药物共同递送的缺陷。因此,需要开发一种新型的基于NO的抑制肿瘤氧气代谢的方法。
精氨酸(Arg)是一种天然的NO前体化合物,这种氨基酸可以被诱导型一氧化氮合酶和/或内源性ROS氧化成NO,这两种物质均在癌细胞中过表达。更重要的是,通过简单的设计与化学合成,Arg可以与各种短肽相连接。这使得Arg拥有了一个显著的优势,即可以通过连接不同的肽序列或保护基团来调节其理化性质,例如疏水性、芳香性堆积和组装特性,以满足不同的制剂需要。
发明内容
针对现有技术存在的缺陷以及Arg分子结构的特点,本发明设计并构建了一种Arg-小肽复合物,通过非共价作用的方式实现光敏剂与Arg的高效共载和共同递送协同杀死肿瘤细胞。
本发明通过以下技术方案实现上述目的:
本发明所述的Arg-小肽复合物与光敏剂共组装纳米粒,是Arg-小肽复合物和光敏剂通过π-π堆积、疏水作用等非共价作用力形成,经过聚乙二醇(PEG)修饰剂修饰或未修饰的共载纳米粒。
所述的Arg-小肽复合物中,小肽分子连接在Arg分子的氨基端和/或羧基端,且在小肽的氮末端和/或碳末端使用或不使用含有芳香基团的化合物进行封端。组成小肽的氨基酸包括但不限于亮氨酸、异亮氨酸、酪氨酸、丙氨酸以及苯丙氨酸,所述含有芳香基团的化合物包括但不限于芴基甲氧羰基(Fmoc)、苄氧羰酰基(Z)、苯环及其同系物、萘环及其同系物和芘环及其同系物。
所述光敏剂包括但不限于母体结构为卟啉环或酞菁环,并在分子内存在酸性基团的光敏剂,所述酸性基团包括但不限于酚羟基、羧基以及磺酸基。
所述的PEG修饰剂为DSPE-PEG、TPGS、PLGA-PEG或PE-PEG,优选DSPE-PEG;所述PEG的分子量为1000、2000或5000,优选2000。
其中,Arg-小肽复合物与光敏剂的质量比为5:1~1:5,优选2:1~1:4,PEG修饰剂的加入量为(Arg-小肽复合物+光敏剂)总质量的0%~50%,优选5%~50%。
进一步地,本发明所述的Arg-小肽复合物与光敏剂共组装纳米粒为:芴基甲氧羰基-亮氨酸-亮氨酸-亮氨酸-精氨酸(Fmoc-L3-Arg)与5,10,15,20-四(4-羟基苯)-21H,23H-卟啉(THPP,CAS号:51094-17-8)形成的共组装纳米粒(THPP/Fmoc-L3-Arg共组装纳米粒),并经过PEG修饰剂修饰。
本发明还提供所述的Arg-小肽复合物与光敏剂共组装纳米粒的制备方法,包括:
将Arg-小肽复合物,光敏剂与PEG修饰剂溶解到有机溶剂中,在搅拌下,将该溶液缓缓滴加到去离子水中,自发形成粒径均一(约80nm)的Arg-小肽复合物与光敏剂共组装纳米粒。
上述制备方法中:
所述的Arg-小肽复合物中,小肽分子连接在Arg分子的氨基端和/或羧基端,且在小肽的氮末端和/或碳末端使用或不使用含有芳香基团的化合物进行封端。组成小肽的氨基酸包括但不限于亮氨酸、异亮氨酸、酪氨酸、丙氨酸以及苯丙氨酸,所述的含有芳香基团的化合物包括但不限于芴基甲氧羰基(Fmoc)、苄氧羰酰基(Z)、苯环及其同系物、萘环及其同系物和芘环及其同系物。
所述光敏剂包括但不限于母体结构为卟啉环或酞菁环,并在分子内存在酸性基团的光敏剂,所述酸性基团包括但不限于酚羟基、羧基以及磺酸基。光敏剂优选为THPP。
所述Arg-小肽复合物与光敏剂的质量比为5:1~1:5,优选2:1~1:4,其中Arg-小肽复合物的浓度范围为2mg/mL~40mg/mL,光敏剂的浓度范围为2mg/mL~40mg/mL;
所述PEG修饰剂为DSPE-PEG、TPGS、PLGA-PEG或PE-PEG,优选DSPE-PEG;所述PEG的分子量为1000、2000或5000,优选2000,PEG修饰剂的加入量为(Arg-小肽复合物+光敏剂)总质量的0%~50%,优选5%~50%;
所述有机溶剂为二甲基亚砜或甲醇或二者的混合溶剂。
本发明所述的Arg-小肽复合物与光敏剂共组装纳米粒在制备抗肿瘤药物中应用,或在制备注射给药、口服给药或局部给药系统中应用。
本发明的有益效果:
(1)采用一步纳米沉淀的方法,制备工艺简单,易于放大生产;
(2)形成的纳米粒子粒径小而均一(约80nm),有利于纳米粒通过渗透与滞留增强(EPR)效应富集于肿瘤部位;
(3)超高的载药量,有利于减小辅料相关的不良反应和毒性;
(4)Arg可在肿瘤细胞内被氧化生成一氧化氮以缓解肿瘤乏氧,与光敏剂产生协同的抗肿瘤效果。
附图说明
图1为本发明应用的THPP、Arg以及Arg-小肽复合物化学结构以及相应沉淀和纳米粒的外观。
图2(A)为本发明实施例1制备的THPP/Fomc-L3-Arg共组装纳米粒的粒径分布,(B)为本发明实施例1制备的THPP/Fomc-L3-Arg共组装纳米粒的透射电子显微镜图。
图3为本发明实施例1中制备的THPP/Fomc-L3-Arg共组装纳米粒在1mol/L的尿素、氯化钠和SDS溶液中粒径变化图。
图4为本发明实施例1中制备的THPP/Fomc-L3-Arg共组装纳米粒4℃放置的粒径-存储时间图。
图5为本发明实施例1中制备的THPP/Fomc-L3-Arg共组装纳米粒在pH 7.4FBS中的粒径-时间图。
图6为本发明实施例1中制备的THPP/Fomc-L3-Arg共组装纳米粒在pH 5.5FBS以及pH5.5H2O2中的粒径-时间图。
图7为本发明实施例9的细胞内一氧化氮水平测定图。
图8为为本发明实施例10为CcO活性测定图
图9为本发明实施例11的细胞培养基氧气含量测定图
图10为本发明实施例12的细胞内活性氧水平测定图;为常氧条件和乏氧条件。
图11为本发明实13的细胞毒性实验结果;(A)为常氧条件,(B)为乏氧条件。
图12为本发明实施例14的共组装纳米粒组织分布图;图A、图B、图C分别对应1h、12h、24h。
图13为本发明为实施例15的在体抗肿瘤实验肿瘤体积变化图(A)和离体肿瘤质量图(B)。
具体实施方式
通过以下具体实例,对本发明的上述内容作进一步的详细说明,但并不表示实施例对本发明的限制。
实施例1:PEG修饰的THPP/Fomc-L3-Arg共组装纳米粒的制备
将一定量的Fomc-L3-Arg、THPP以及PEG修饰剂DSPE-PEG2000分别溶解于一定量的二甲基亚砜中,配制成20.0mg/mL Fomc-L3-Arg储备液、20.0mg/mL THPP储备液和5.0mg/mLDSPE-PEG2000储备液。将25μL Fomc-L3-Arg储备液、75μL THPP储备液和120μL DSPE-PEG2000储备液进行混合(Fomc-L3-Arg和光敏剂的质量比为1:3),在搅拌速度800rpm条件下,将混合溶液滴加到2mL去离子水中使其自发形成纳米粒。将制剂(即自发形成的纳米粒)转移至超滤管中3000rpm离心15min以除去制剂中的有机溶剂,加去离子水定容。
使用透射电子显微镜观测本实施例中制备的PEG修饰THPP/Fomc-L3-Arg共组装纳米粒的粒径和形态,结果见图2B;使用动态光散射法测量PEG修饰THPP/Fomc-L3-Arg共组装纳米粒的粒径,粒径分布见图2A。结果表明,PEG修饰THPP/Fomc-L3-Arg共组装纳米粒粒径约80nm,呈均一球形。
实施例2:PEG修饰的THPP/Fomc-L3-Arg共组装纳米粒的制备
将一定量的THPP、Fomc-L3-Arg以及PEG修饰剂DSPE-PEG2000分别溶解于一定量的甲醇中,配制成10.0mg/mL Fomc-L3-Arg储备液、10.0mg/mL THPP储备液和2.5mg/mL DSPE-PEG2000储备液。将20μL Fomc-L3-Arg储备液、20μL THPP储备液和80μL DSPE-PEG2000储备液进行混合(Fomc-L3-Arg和光敏剂的质量比为1:1),在搅拌速度800rpm条件下,将混合溶液滴加到10mL去离子水中使其自发形成纳米粒。将制剂转移至超滤管中2000rpm离心15min以除去制剂中的有机溶剂,加去离子水定容至所需浓度即可。使用透射电子显微镜观测本实施例中制备的PEG修饰THPP/Fomc-L3-Arg共组装纳米粒的粒径和形态。本实施例中得到的PEG修饰的THPP/Fomc-L3-Arg共组装纳米粒粒径约80nm,呈均一球形。
实施例3:PEG修饰的THPP/Fomc-L3-Arg共组装纳米粒的制备
将一定量的THPP、Fomc-L3-Arg以及PEG修饰剂DSPE-PEG2000分别溶解于一定量的甲醇中,配制成40.0mg/mL Fomc-L3-Arg储备液、20.0mg/mL THPP储备液和2.5mg/mL DSPE-PEG2000储备液。将10μL Fomc-L3-Arg储备液、10μL THPP储备液和24μL DSPE-PEG2000储备液进行混合(Fomc-L3-Arg和光敏剂的质量比为2:1),在搅拌速度800rpm条件下,将混合溶液滴加到10mL去离子水中使其自发形成纳米粒。将制剂转移至超滤管中2000rpm离心15min以除去制剂中的有机溶剂,加去离子水定容至所需浓度即可。使用透射电子显微镜观测本实施例中制备的PEG修饰THPP/Fomc-L3-Arg共组装纳米粒的粒径和形态。本实施例中得到的PEG修饰的THPP/Fomc-L3-Arg共组装纳米粒粒径约80nm,呈均一球形。
实施例4:THPP与Fmoc-L3-Arg组装机制的考察
参考实施例1中THPP/Fmoc-L3-Arg NPs的制备方法,尝试用Fmoc-L3-Arg的分子片段包括Fmoc-Arg、Fmoc-L-L-L、L-L-L-Arg与THPP共组。此外,将实施例1中制备的THPP/Fmoc-L3-Arg NPs分别放入1mol/L的尿素、氯化钠和SDS溶液中,并在37℃恒温振荡器(100rpm)中孵育,于孵育0.5、1以及2h后使用粒径仪测定纳米粒粒径。
结果如图1和图3所示。THPP/Fomc-L3-Arg NPs在SDS溶液中孵育后,其粒径发生显着变化,结合THPP分子在水中会析出大量沉淀的现象,这说明两个分子之间的疏水相互作用。此外,芳香性基团如Fmoc、苯衍生物和卟啉环之间形成的π-π堆积作用在共组装中也具有重要作用。因此,不具有芳香基团的L-L-L-Arg无法与THPP共组装。进一步的,尝试使用Fomc-L-L-L-OH与THPP共组装时,依旧观察到了大量沉淀物。该结果说明Fomc-L3-Arg中的Arg残基促进了共组装过程的发生,推测是其碱性胍基与THPP的酸性的酚羟基形成了电荷吸引作用。如图3所示的THPP/Fomc-L3-Arg NPs在氯化钠溶液中显著的解离也进一步证实了THPP和Fomc-L3-Arg之间存在电荷吸引作用。在尿素溶液中孵育后粒径显著增加,表明THPP和Fomc-L3-Arg两种分子间存在氢键。氢键推测是由Fmoc-L3-Arg中的羰基和THPP分子中的氢原子(NH)形成的。因此,只有一个羰基的Fomc-Arg无法与THPP共组装,形成了大量沉淀物。总结上述实验结果,静电引力、π-π堆积、疏水相互作用以及氢键作用力均是THPP/Fomc-L3-Arg NPs形成的主要作用力。
实施例5:THPP/Fomc-L3-Arg共组装纳米粒的4℃存储稳定性试验
将实施例1中制备的HPP/Fomc-L3-Arg共组装纳米粒在4℃的条件下放置,并且在预定的时间点(0,1,3,5,7天)通过动态光散射法测定其粒径变化。结果如图4所示,共组装纳米粒在7天内粒径无明显变化,具有良好的4℃存储稳定性。
实施例6:THPP/Fomc-L3-Arg共组装纳米粒的FBS稳定性试验
将实施例1中制备的HPP/Fomc-L3-Arg共组装纳米粒在37℃的条件下在pH 7.4的10%PBS溶液中孵育24h,并且在预定的时间点(0,2,4,8,12,24h)通过动态光散射法测定其粒径变化。结果如图5所示,共组装纳米粒在24小时内粒径无明显变化,显示出良好的稳定性。
实施例7:THPP/Fomc-L3-Arg共组装纳米粒的酸敏感性考察
共组装纳米粒不仅需要在储存和生理条件下稳定,而且应在肿瘤环境中响应分解以释放药物。因此,通过监测THPP/Fomc-L3-Arg共组装纳米粒粒径在模拟肿瘤微环境的介质中的变化来评估纳米颗粒的肿瘤响应性。与生理条件下的pH7.4不同,肿瘤组织在间隙(pH 6.5-7.2),细胞内体(intracellular endosomes)(pH 5.0-6.5)和溶酶体(pH 4.5-5.0)中呈现弱酸环境。因此,将实施例1中制备的THPP/Fomc-L3-Arg共组装纳米粒在37℃的条件下pH 5.5的PBS中孵育24h,并且在预定的时间点(0,2,4,8,12,24h)通过动态光散射法测定其粒径。结果如图6所示,纳米粒在24小时内粒径逐渐增加,推测是由于酸性条件下过多的氢离子质子化了精氨酸分子中的胍基,使得THPP与Fomc-L3-Arg分子间的电荷作用力变弱所导致的。
实施例8:THPP/Fomc-L3-Arg共组装纳米粒的氧化敏感性考察
除了弱酸性环境外,肿瘤细胞具有很高的ROS水平。Arg中的胍基被ROS氧化后会产生弱碱性的酰胺基,使得Fomc-L3-Arg与THPP之间的电荷相互作用减弱。因此,本实施例中将实施例1中制得的THPP/Fomc-L3-Arg共组装纳米粒在37℃的条件下的pH5.5 PBS+10mMH2O2中孵育。结果如图6所示,纳米粒在此介质中具有较强敏感性,孵育一小时后便完全解离并析出,这表明其在肿瘤细胞内可能具有较强的响应性。
实施例9:THPP、Fomc-L3-Arg、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒细胞内一氧化氮生成量测定
采用DAF-FM DA荧光探针检测THPP/Fmoc-L3-Arg共组装纳米粒在4T1细胞中NO的产生量。检测原理如下:DAF-FM DA进入细胞后被细胞内的酯酶催化形成不能穿过细胞膜的DAF-FM,DAF-FM本身仅有很弱的荧光,但与NO反应后会生成具有强烈绿色荧光的荧光素-苯骈三氮唑,借此测定细胞内NO水平(激发和发射波长分别为495nm和515nm)。
将4T1细胞按照每孔1×105(1mL)个细胞的密度接种到12孔板中,放入培养箱中12h使细胞贴壁后弃去板中旧培养基,加入含有THPP溶液剂、Fmoc-L3-Arg溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒的新鲜培养基(THPP浓度为15μg/mL,Fmoc-L3-Arg浓度为5μg/mL),并放置于培养箱中孵育4h。设置3个平行孔,使用空白培养基培养的细胞作为对照组。孵育结束后,将含药培养基倒掉,使用冰浴预冷的PBS清洗细胞三次,加入含有DAF-FM DA荧光探针的新鲜培养基并放置于培养箱中继续孵育30min。孵育结束后,使用冰浴预冷的PBS清洗细胞三次,之后使用胰酶将细胞消化下来,加入预冷PBS终止消化,离心,弃去上清液并复悬于PBS中,采用流式细胞仪(通道1)测定每个样品中荧光素-苯骈三氮唑的荧光强度,细胞采集数目104个。
结果如图7所示,THPP溶液剂对细胞本身的NO水平并没有影响,Fmoc-L3-Arg溶液剂以及THPP/Fmoc-L3-Arg物理混合溶液剂显著提高了细胞内的NO水平,说明Fmoc-L3-Arg在进入细胞后可以快速的代谢生成NO。相比溶液剂,THPP/Fmoc-L3-Arg共组装纳米粒在细胞内的NO产量稍低,推测是纳米粒摄取效率较低,进入细胞内的Fmoc-L3-Arg量稍低导致的。
实施例10:考查THPP、Fomc-L3-Arg、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒对CcO活性影响
采用线粒体呼吸链复合体IV(CcO)活性检测试剂盒测定CcO活性。将4T1细胞按照每孔5×105(5mL)个细胞的密度接种到12孔板中,放入培养箱中12h使细胞贴壁后弃去板中旧培养基,加入含有THPP溶液剂、Fmoc-L3-Arg溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒的新鲜培养基(THPP浓度为15μg/mL,Fmoc-L3-Arg浓度为5μg/mL),并放置于培养箱中孵育4h。设置3个平行孔,使用未加药培养基培养的细胞作为对照组。孵育结束后每孔加入0.1mL提取液,之后按照检测试剂盒说明书上的步骤测定CcO活性。
结果如图8所示,THPP溶液剂对CcO的活性并没有显著影响,Fmoc-L3-Arg溶液剂以及THPP/Fmoc-L3-Arg物理混合溶液剂共孵育的细胞CcO活性明显下降,而THPP/Fmoc-L3-Arg共组装纳米粒对线粒呼吸链的抑制能力较弱,推测是纳米粒在细胞内NO产量较低导致的。
实施例11:THPP、Fomc-L3-Arg、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒缓解细胞乏氧考察
通过测定细胞培养基中氧气(O2)含量间接反映细胞内氧含量,为了避免培养箱中的氧气不断溶解于培养基中造成测定干扰,加药后我们将细胞置于密封的培养盒中进行孵育,并在孵育后尽快的测定培养基中氧气浓度。将4T1细胞按照每孔106(3mL)个细胞的密度接种到6孔板中,放入培养箱中12h使细胞贴壁后弃去板中旧培养基。加入5mL含有THPP溶液剂、Fmoc-L3-Arg溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒的新鲜培养基(THPP浓度为15μg/mL,Fmoc-L3-Arg浓度为5μg/mL)。之后将细胞放入密封的低氧培养盒并在37℃的条件下孵育4h。使用在常氧条件下、空白培养基培养的细胞作为对照,孵育结束后立刻使用便携式溶解氧测定仪测定细胞培养基中氧气浓度。
结果如图9所示,THPP溶液剂对肿瘤细胞的氧气消耗没有显著影响,Fmoc-L3-Arg溶液剂以及THPP/Fmoc-L3-Arg物理混合溶液剂可以有效抑制CcO活性因此明显降低了细胞氧气消耗量。相比之下,THPP/Fmoc-L3-Arg共组装纳米粒抑制CcO活性的能力稍弱,因此抑制细胞氧气代谢的能力稍弱。
实施例12:THPP、Fomc-L3-Arg、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒细胞内ROS产量考察
采用DCFH-DA探针检测THPP/Fmoc-L3-Arg共组装纳米粒在4T1细胞中ROS的产生量。检测原理如下:DCFH-DA本身不具有荧光,其进入细胞后会被细胞中的酯酶水解生成DCFH。ROS可以特异性的氧化DCFH生成具有荧光信号的DCF,借此测定细胞内ROS水平(激发和发射波长分别为504nm和529nm)。
将4T1细胞按照每孔105(1mL)个细胞的密度接种到12孔板中,放入培养箱中12h使细胞贴壁后弃去板中旧培养基,加入1mL含有THPP溶液剂、Fmoc-L3-Arg溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒的新鲜培养基(THPP浓度为1μg/mL,Fmoc-L3-Arg浓度为0.33μg/mL),并放置于培养箱中常氧和密封乏氧各自孵育4h。设置3个平行孔,用空白培养基孵育的细胞作为对照组。孵育结束后,向每孔含药培养基加入0.1mL含有100μg/mL DCFH-DA的培养基母液并放置于培养箱中常氧和乏氧各自孵育30min。孵育结束后,不光照或将其置于波长660nm、功率40mW cm-2的光源下照射2min,使用胰酶将细胞消化下来,加入预冷PBS终止消化,离心,弃去上清液并复悬于PBS中,采用流式细胞仪(通道1)测定每个样品中DCF的荧光强度,细胞采集数目104个。
结果如图10所示,在常氧条件下,肿瘤细胞具有充足的氧气供应,THPP溶液剂与THPP/Fmoc-L3-Arg物理混合溶液剂的ROS的水平并没有显著差异,THPP/Fmoc-L3-Arg共组装纳米粒的摄取效率低于溶液剂,因此ROS产量稍低。在乏氧条件下,所有给药组细胞内的ROS产量均明显下降,Fmoc-L3-Arg可以显著的提高肿瘤内的氧气含量,因此THPP/Fmoc-L3-Arg物理混合溶液剂的ROS产量明显高于THPP溶液剂的ROS产量。此外,尽管THPP/Fmoc-L3-Arg共组装纳米粒的摄取效率略低于THPP溶液剂,但共载的Fmoc-L3-Arg有效的缓解肿瘤乏氧,因此其ROS产量高于THPP溶液剂。
实施例13:THPP、Fomc-L3-Arg、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒细胞毒性考察
采用MTT法测定THPP溶液剂、Fmoc-L3-Arg溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒对在常氧以及乏氧条件下对4T1细胞的毒性。向96孔细胞培养板中每孔接种1000个4T1细胞(100μL),将其置于细胞培养箱中孵育12h使细胞贴壁。弃去旧培养基后加入由新鲜细胞培养基稀释的不同浓度的THPP溶液剂、Fmoc-L3-Arg溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒。每个浓度设置三个平行孔,空白对照组加不含药的新鲜培养基。常氧或密封乏氧孵育4h后,将上述96孔板置于660nm、功率40mW cm-2的光源下照射2min后放回。孵育24h后,将所有96孔板取出,向每孔中加入20μL MTT溶液并在培养箱中孵育4h,甩板并吸干残留培养基,每孔加入200μLDMSO并置于振荡器上振荡10min以溶解蓝紫色结晶物。使用多功能酶标仪在570nm处测定各孔矫正零后的紫外吸光度值并计算细胞存活率。通过Graphpad prism软件计算半数抑制浓度(IC50值)。
结果如图11(A)所示,常氧条件下具有充足的氧气供应,THPP/Fmoc-L3-Arg物理混合溶液剂与THPP溶液剂的ROS产量并没有显著差异,因此混合溶液剂并没有显示出更强的细胞毒。相比于THPP溶液剂,THPP/Fmoc-L3-Arg共组装纳米粒的细胞摄取效率稍低,因此细胞毒稍弱于THPP溶液剂。常氧条件下各制剂IC50值分别为:THPP溶液(56.91ng/mL)≈THPP/Fomc-L3-Arg物理混合物(53.81ng/mL)<THPP/Fomc-L3-Arg NP(83.62ng/mL)。
如图11(B)所示,在乏氧条件下,细胞的氧气供给不足,所有给药组的ROS产量均明显下降,细胞毒也因此变弱。Fmoc-L3-Arg可以抑制肿瘤细胞自身的氧气代谢、缓解肿瘤乏氧,因此THPP/Fmoc-L3-Arg物理混合溶液剂的ROS产量明显高于THPP溶液剂,细胞毒也明显增强。此外,在Fmoc-L3-Arg的帮助下,纳米粒的细胞毒也略高于THPP溶液剂。乏氧条件下各制剂IC50值分别为:THPP/Fomc-L3-Arg混合物(111.3ng/mL)<THPP/Fomc-L3-Arg共组装纳米粒(146.4ng/mL)<THPP溶液(189.1ng/mL)。
实施例14:THPP、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒组织分布考察
用适量胰酶将4T1细胞消化,加入RPMI 1640培养液终止消化后离心,弃去上清液后加入PBS(pH 7.4)将细胞均匀分散成5×106cells/mL的细胞悬液,放入冰浴中备用。将BALB/c小鼠固定后,用胰岛素注射器吸取100μL细胞悬液接种于小鼠右后侧腰背部皮下。待肿瘤体积生长至400mm3时,将小鼠随机分为3组,分别尾静脉注射THPP溶液剂、THPP/Fmoc-L3-Arg物理混合溶液剂以及THPP/Fmoc-L3-Arg共组装纳米粒(THPP等效剂量:5mg/kg,Fmoc-L3-Arg等效剂量:1.66mg/kg)。分别在给药后1h、12h以及24h脱臼处死老鼠,取出心、肝、脾、肺、肾以及肿瘤组织,用生理食盐水清洗血迹并擦干后,将组织置于小动物活体成像仪中对各个器官和肿瘤中的荧光信号强度进行分析,并对其体内分布进行半定量分析。
结果如图12所示,THPP溶液剂在肿瘤组织中的蓄积较弱,肿瘤组织处的荧光从给药后1h至24h不断变弱,THPP/Fmoc-L3-Arg物理混合溶液剂也未具有比单一THPP溶液剂更强的肿瘤蓄积能力,由于实体瘤具有“增强渗透与滞留效应(EPR effect)”,因此THPP/Fmoc-L3-Arg共组装纳米粒显示出了较强的肿瘤靶向能力,肿瘤组织处的荧光从给药后1h至12h变强,且在给药后12h时最强。
实施例15:THPP、THPP/Fomc-L3-Arg物理混合物、THPP/Fomc-L3-Arg共组装纳米粒抑制肿瘤组织生长能力考察
用适量胰酶将4T1细胞消化,加入RPMI 1640培养液终止消化后离心,弃去上清液后加入PBS(pH 7.4)将细胞均匀分散成5×106cells/mL的细胞悬液,放入冰浴中备用。将BALB/c小鼠固定后,用胰岛素注射器吸取100μL细胞悬液并将其接种于小鼠右后侧腰背部皮下。待肿瘤体积生长至约100mm3时,将小鼠随机分为5组,分别为PBS组、Fmoc-L3-Arg溶液剂组、THPP溶液剂+光照组、THPP/Fmoc-L3-Arg物理混合溶液剂+光照组以及THPP/Fmoc-L3-Arg共组装纳米粒+光照组(THPP等效剂量:5mg/kg,Fmoc-L3-Arg等效剂量1.66mg/kg)。根据图12药物组织分布结果,THPP溶液剂以及THPP/Fmoc-L3-Arg混合溶液剂于给药后1h、THPP/Fmoc-L3-Arg共组装纳米粒于给药后12h给予波长660nm、功率密度150mW/cm的光源照射5min。每2天给药一次,共给药4次,每天测量和记录荷瘤小鼠的体重与肿瘤体积,体积计算公式为(长径×短径2×1/2)。在第10天将小鼠处死,剥离肿瘤组织,拍照称重。
药效学结果如图13所示,PBS组小鼠的肿瘤生长迅速,在给药后的第10天生长到体积约800mm3。THPP溶液剂在肿瘤组织蓄积能力较差,因此抑制小鼠体内肿瘤组织生长的能力较弱;与THPP溶液剂相比,THPP/Fmoc-L3-Arg物理混合溶液剂的药效并未显著提高,这说Fmoc-L3-Arg分子无法与THPP分子同时蓄积到肿瘤组织发挥协同治疗效果。相比于上述几种治疗方案,THPP/Fmoc-L3-Arg共组装纳米粒经激发光照射后有效的抑制了肿瘤组织的生长,这主要归功于共组装纳米粒良好的制剂学性质(1)具有良好的胶体稳定性;(2)可通过EPR效应高效的蓄积在肿瘤中组织;(3)被细胞摄取后可快速释放THPP,同时Fmoc-L3-Arg可生产NO有效缓解肿瘤细胞乏氧,提高光敏剂ROS产量以及PDT疗效。药效实验终止后,肿瘤组织的重量也进一步表明THPP/Fmoc-L3-Arg NPs具有良好的抗肿瘤效果。
Claims (10)
1.一种精氨酸-小肽复合物与光敏剂共组装纳米粒,其特征在于,其为精氨酸-小肽复合物与光敏剂通过非共价作用力形成,经过PEG修饰剂修饰或未修饰的共载纳米粒;
所述的精氨酸-小肽复合物中,小肽分子连接在精氨酸分子的氨基端和/或羧基端,且在小肽的氮末端和/或碳末端使用或不使用含有芳香基团的化合物进行封端;
所述的光敏剂包括但不限于母体结构为卟啉环或酞菁环,并在分子内存在酸性基团的光敏剂,所述的酸性基团包括但不限于酚羟基、羧基以及磺酸基;
所述的非共价作用力包括π-π堆积、疏水作用;
所述的PEG修饰剂为DSPE-PEG、TPGS、PLGA-PEG或PE-PEG;所述PEG的分子量为1000、2000或5000。
2.根据权利要求1所述的精氨酸-小肽复合物与光敏剂共组装纳米粒,其特征在于,精氨酸-小肽复合物与光敏剂的质量比为5:1~1:5,PEG修饰剂的加入量为精氨酸-小肽复合物和光敏剂总质量的0%~50%;组成小肽的氨基酸包括但不限于亮氨酸、异亮氨酸、酪氨酸、丙氨酸以及苯丙氨酸;所述的含有芳香基团的化合物包括但不限于芴基甲氧羰基、苄氧羰酰基、苯环及其同系物、萘环及其同系物和芘环及其同系物。
3.根据权利要求1或2所述的精氨酸-小肽复合物与光敏剂共组装纳米粒,其特征在于,所述的精氨酸-小肽复合物与光敏剂共组装纳米粒为:芴基甲氧羰基-亮氨酸-亮氨酸-亮氨酸-精氨酸与5,10,15,20-四(4-羟基苯)-21H,23H-卟啉形成的共组装纳米粒,并经过PEG修饰剂修饰。
4.根据权利要求3所述的精氨酸-小肽复合物与光敏剂共组装纳米粒,其特征在于,精氨酸-小肽复合物与光敏剂的质量比为2:1~1:4,PEG修饰剂为DSPE-PEG,分子量为2000,PEG修饰剂的加入量为精氨酸-小肽复合物和光敏剂总质量的5%~50%。
5.一种权利要求1所述的精氨酸-小肽复合物与光敏剂共组装纳米粒的制备方法,其特征在于,将精氨酸-小肽复合物,光敏剂与PEG修饰剂溶解到有机溶剂中,在搅拌下,将该溶液缓缓滴加到去离子水中,自发形成粒径均一球形精氨酸-小肽复合物与光敏剂共组装纳米粒。
6.根据权利要求5所述的精氨酸-小肽复合物与光敏剂共组装纳米粒的制备方法,其特征在于,所述的精氨酸-小肽复合物中,小肽分子连接在精氨酸分子的氨基端和/或羧基端,且在小肽的氮末端和/或碳末端使用或不使用含有芳香基团的化合物进行封端;组成小肽的氨基酸包括但不限于亮氨酸、异亮氨酸、酪氨酸、丙氨酸以及苯丙氨酸,所述的含有芳香基团的化合物包括但不限于芴基甲氧羰基、苄氧羰酰基、苯环及其同系物、萘环及其同系物和芘环及其同系物;
所述光敏剂包括但不限于母体结构为卟啉环或酞菁环,并在分子内存在酸性基团的光敏剂,所述酸性基团包括但不限于酚羟基、羧基以及磺酸基;
所述精氨酸-小肽复合物与光敏剂的质量比为5:1~1:5,其中精氨酸-小肽复合物的浓度范围为2mg/mL~40mg/mL,光敏剂的浓度范围为2mg/mL~40mg/mL;
所述PEG修饰剂为DSPE-PEG、TPGS、PLGA-PEG或PE-PEG;所述PEG的分子量为1000、2000或5000,PEG修饰剂的加入量为精氨酸-小肽复合物和光敏剂总质量的0%~50%。
7.根据权利要求5所述的精氨酸-小肽复合物与光敏剂共组装纳米粒的制备方法,其特征在于,所述有机溶剂为二甲基亚砜或甲醇或二者的混合溶剂。
8.根据权利要求5所述的精氨酸-小肽复合物与光敏剂共组装纳米粒的制备方法,其特征在于,所述光敏剂为5,10,15,20-四(4-羟基苯)-21H,23H-卟啉。
9.根据权利要求6所述的精氨酸-小肽复合物与光敏剂共组装纳米粒的制备方法,其特征在于,所述精氨酸-小肽复合物与光敏剂的质量比为2:1~1:4,所述PEG修饰剂为DSPE-PEG,所述PEG的分子量为2000,PEG修饰剂的加入量为精氨酸-小肽复合物和光敏剂总质量的5%~50%。
10.一种权利要求1所述的精氨酸-小肽复合物与光敏剂共组装纳米粒的应用,其特征在于,在制备抗肿瘤药物中应用,或在制备注射给药、口服给药或局部给药系统中应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111068594.6A CN113797336B (zh) | 2021-09-13 | 2021-09-13 | 精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111068594.6A CN113797336B (zh) | 2021-09-13 | 2021-09-13 | 精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113797336A true CN113797336A (zh) | 2021-12-17 |
CN113797336B CN113797336B (zh) | 2023-06-16 |
Family
ID=78940891
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111068594.6A Active CN113797336B (zh) | 2021-09-13 | 2021-09-13 | 精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113797336B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114404364A (zh) * | 2022-01-18 | 2022-04-29 | 广西大学 | 一种可诱导肿瘤相关巨噬细胞m1型极化的光敏纳米胶束的构建及其抗肿瘤应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100075899A1 (en) * | 2006-09-28 | 2010-03-25 | Juan Chen | Targeted photodynamic therapy agent |
WO2021036752A1 (zh) * | 2019-08-23 | 2021-03-04 | 国家纳米科学中心 | 一种靶向乏氧肿瘤的短肽小分子自组装纳米材料及其制备方法和应用 |
CN112843247A (zh) * | 2021-01-19 | 2021-05-28 | 南开大学 | 一种具有线粒体靶向性的多肽超分子Bcl-xL拮抗剂纳米药物的制备方法 |
-
2021
- 2021-09-13 CN CN202111068594.6A patent/CN113797336B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100075899A1 (en) * | 2006-09-28 | 2010-03-25 | Juan Chen | Targeted photodynamic therapy agent |
WO2021036752A1 (zh) * | 2019-08-23 | 2021-03-04 | 国家纳米科学中心 | 一种靶向乏氧肿瘤的短肽小分子自组装纳米材料及其制备方法和应用 |
CN112843247A (zh) * | 2021-01-19 | 2021-05-28 | 南开大学 | 一种具有线粒体靶向性的多肽超分子Bcl-xL拮抗剂纳米药物的制备方法 |
Non-Patent Citations (1)
Title |
---|
QIN WANG等: "Synthesis of fluorescent nanoprobe with simultaneous response to intracellular pH and Zn2+ for tumor cell distinguishment", 《MICROCHIMICA ACTA》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114404364A (zh) * | 2022-01-18 | 2022-04-29 | 广西大学 | 一种可诱导肿瘤相关巨噬细胞m1型极化的光敏纳米胶束的构建及其抗肿瘤应用 |
CN114404364B (zh) * | 2022-01-18 | 2023-06-16 | 广西大学 | 一种可诱导肿瘤相关巨噬细胞m1型极化的光敏纳米胶束的构建及其抗肿瘤应用 |
Also Published As
Publication number | Publication date |
---|---|
CN113797336B (zh) | 2023-06-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Zhang et al. | Positive feedback nanoamplifier responded to tumor microenvironments for self-enhanced tumor imaging and therapy | |
Wu et al. | Recent progress in the augmentation of reactive species with nanoplatforms for cancer therapy | |
Yang et al. | An oxygen self-sufficient NIR-responsive nanosystem for enhanced PDT and chemotherapy against hypoxic tumors | |
CN109718207B (zh) | 化疗药-光敏剂共组装纳米粒及其构建 | |
Yuan et al. | Mitochondria targeted O2 economizer to alleviate tumor hypoxia for enhanced photodynamic therapy | |
Wang et al. | Enhancing selective photosensitizer accumulation and oxygen supply for high-efficacy photodynamic therapy toward glioma by 5-aminolevulinic acid loaded nanoplatform | |
CN110743012A (zh) | 一种葡萄糖氧化酶修饰的介孔二氧化锰药物组合物的制备方法及应用 | |
CN113521098B (zh) | 铂(Ⅳ)及cRGD修饰的GA/Fe纳米颗粒搭载多柔比星及其靶向治疗肿瘤的方法 | |
Yin et al. | Hypoxia-alleviated sonodynamic therapy based on a hybrid protein oxygen carrier to enhance tumor inhibition | |
CN108658995B (zh) | 一种二硫联吡啶修饰锌酞菁及其制备方法和应用 | |
Yin et al. | Synergistically enhanced multienzyme catalytic nanoconjugates for efficient cancer therapy | |
CN111135299A (zh) | 光敏剂-低氧激活前药一体化前药自组装纳米粒的构建 | |
CN113018267A (zh) | 不饱和脂肪酸-光敏剂共组装纳米粒及其构建方法和应用 | |
Zheng et al. | A chemical biology toolbox to overcome the hypoxic tumor microenvironment for photodynamic therapy: a review | |
CN113648401B (zh) | 一种蛋白酶体抑制增敏光动力治疗的杂化纳米组装体及其制备与应用 | |
CN113797336B (zh) | 精氨酸-小肽复合物与光敏剂共组装纳米粒及其制备方法和应用 | |
US20200222564A1 (en) | Compound Amphiphilic Peptide Nanomicelle, Preparation and Use Thereof | |
CN114191550A (zh) | 一种自携氧纳米光敏制剂及其制备方法和应用 | |
Yu et al. | Ultra-stable MOF@ MOF nanoplatform for photodynamic therapy sensitized by relieved hypoxia due to mitochondrial respiration inhibition | |
CN110755637B (zh) | 谷胱甘肽抑制剂-光敏剂共组装纳米粒及其构建 | |
CN106606783B (zh) | 一种靶向共递释光敏剂与化疗药物的药物递释系统 | |
Li et al. | Palliating the escalated post-PDT tumor hypoxia with a dual cascade oxygenation nanocomplex | |
Liu et al. | Modulation of hypoxia and redox in the solid tumor microenvironment with a catalytic nanoplatform to enhance combinational chemodynamic/sonodynamic therapy | |
CN115364235A (zh) | 一种锌离子驱动氧气节约和基因沉默的生物活性纳米载体及其制备方法和应用 | |
CN109806404B (zh) | 一种能够双级递氧的白蛋白纳米粒及其制备方法与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |