CN114306609A - 一种钙超载型小分子药物自组装纳米粒子及其应用 - Google Patents
一种钙超载型小分子药物自组装纳米粒子及其应用 Download PDFInfo
- Publication number
- CN114306609A CN114306609A CN202210114493.6A CN202210114493A CN114306609A CN 114306609 A CN114306609 A CN 114306609A CN 202210114493 A CN202210114493 A CN 202210114493A CN 114306609 A CN114306609 A CN 114306609A
- Authority
- CN
- China
- Prior art keywords
- calcium
- nanoparticles
- self
- small molecule
- molecule drug
- 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
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 67
- 230000001964 calcium overload Effects 0.000 title claims abstract description 26
- 239000003814 drug Substances 0.000 title claims abstract description 24
- 229940079593 drug Drugs 0.000 title claims abstract description 23
- 238000001338 self-assembly Methods 0.000 title claims abstract description 20
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims abstract description 50
- 229940109262 curcumin Drugs 0.000 claims abstract description 25
- 235000012754 curcumin Nutrition 0.000 claims abstract description 25
- 239000004148 curcumin Substances 0.000 claims abstract description 25
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 claims abstract description 25
- YKPUWZUDDOIDPM-SOFGYWHQSA-N capsaicin Chemical group COC1=CC(CNC(=O)CCCC\C=C\C(C)C)=CC=C1O YKPUWZUDDOIDPM-SOFGYWHQSA-N 0.000 claims abstract description 18
- 102000003566 TRPV1 Human genes 0.000 claims abstract description 17
- 101150016206 Trpv1 gene Proteins 0.000 claims abstract description 17
- 239000000556 agonist Substances 0.000 claims abstract description 17
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 14
- 229960002504 capsaicin Drugs 0.000 claims abstract description 12
- 239000003381 stabilizer Substances 0.000 claims abstract description 11
- -1 capsaicine ester Chemical class 0.000 claims abstract description 10
- 235000017663 capsaicin Nutrition 0.000 claims abstract description 9
- IKYCZSUNGFRBJS-UHFFFAOYSA-N Euphorbia factor RL9 = U(1) = Resiniferatoxin Natural products COC1=CC(O)=CC(CC(=O)OCC=2CC3(O)C(=O)C(C)=CC3C34C(C)CC5(OC(O4)(CC=4C=CC=CC=4)OC5C3C=2)C(C)=C)=C1 IKYCZSUNGFRBJS-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229930003316 Vitamin D Natural products 0.000 claims abstract description 6
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 claims abstract description 6
- FBUBVLUPUDBFME-UHFFFAOYSA-N Xanthoxylin Chemical compound COC1=CC(O)=C(C(C)=O)C(OC)=C1 FBUBVLUPUDBFME-UHFFFAOYSA-N 0.000 claims abstract description 6
- NLDDIKRKFXEWBK-AWEZNQCLSA-N gingerol Chemical compound CCCCC[C@H](O)CC(=O)CCC1=CC=C(O)C(OC)=C1 NLDDIKRKFXEWBK-AWEZNQCLSA-N 0.000 claims abstract description 6
- JZLXEKNVCWMYHI-UHFFFAOYSA-N gingerol Natural products CCCCC(O)CC(=O)CCC1=CC=C(O)C(OC)=C1 JZLXEKNVCWMYHI-UHFFFAOYSA-N 0.000 claims abstract description 6
- 235000002780 gingerol Nutrition 0.000 claims abstract description 6
- DSDNAKHZNJAGHN-UHFFFAOYSA-N resinferatoxin Natural products C1=C(O)C(OC)=CC(CC(=O)OCC=2CC3(O)C(=O)C(C)=CC3C34C(C)CC5(OC(O4)(CC=4C=CC=CC=4)OC5C3C=2)C(C)=C)=C1 DSDNAKHZNJAGHN-UHFFFAOYSA-N 0.000 claims abstract description 6
- DSDNAKHZNJAGHN-MXTYGGKSSA-N resiniferatoxin Chemical compound C1=C(O)C(OC)=CC(CC(=O)OCC=2C[C@]3(O)C(=O)C(C)=C[C@H]3[C@@]34[C@H](C)C[C@@]5(O[C@@](O4)(CC=4C=CC=CC=4)O[C@@H]5[C@@H]3C=2)C(C)=C)=C1 DSDNAKHZNJAGHN-MXTYGGKSSA-N 0.000 claims abstract description 6
- 229940073454 resiniferatoxin Drugs 0.000 claims abstract description 6
- 235000019166 vitamin D Nutrition 0.000 claims abstract description 6
- 239000011710 vitamin D Substances 0.000 claims abstract description 6
- 150000003710 vitamin D derivatives Chemical class 0.000 claims abstract description 6
- 229940046008 vitamin d Drugs 0.000 claims abstract description 6
- OJYLAHXKWMRDGS-UHFFFAOYSA-N zingerone Chemical compound COC1=CC(CCC(C)=O)=CC=C1O OJYLAHXKWMRDGS-UHFFFAOYSA-N 0.000 claims abstract description 6
- OQWKEEOHDMUXEO-UHFFFAOYSA-N (6)-shogaol Natural products CCCCCC=CC(=O)CCC1=CC=C(O)C(OC)=C1 OQWKEEOHDMUXEO-UHFFFAOYSA-N 0.000 claims abstract description 3
- KAWOEDMUUFFXAM-UHFFFAOYSA-N CC1(C)CCCC2(C)C(C)C(C=O)=CCC21 Polymers CC1(C)CCCC2(C)C(C)C(C=O)=CCC21 KAWOEDMUUFFXAM-UHFFFAOYSA-N 0.000 claims abstract description 3
- HMXRXBIGGYUEAX-SFHVURJKSA-N Evodiamine Natural products CN1[C@H]2N(CCc3[nH]c4ccccc4c23)C(=O)c5ccccc15 HMXRXBIGGYUEAX-SFHVURJKSA-N 0.000 claims abstract description 3
- TXDUTHBFYKGSAH-SFHVURJKSA-N Evodiamine Chemical compound C1=CC=C2N(C)[C@@H]3C(NC=4C5=CC=CC=4)=C5CCN3C(=O)C2=C1 TXDUTHBFYKGSAH-SFHVURJKSA-N 0.000 claims abstract description 3
- AZJUJOFIHHNCSV-KCQAQPDRSA-N Polygodial Polymers C[C@@]1([C@H](C(C=O)=CC2)C=O)[C@@H]2C(C)(C)CCC1 AZJUJOFIHHNCSV-KCQAQPDRSA-N 0.000 claims abstract description 3
- OQWKEEOHDMUXEO-BQYQJAHWSA-N [6]-Shogaol Chemical compound CCCCC\C=C\C(=O)CCC1=CC=C(O)C(OC)=C1 OQWKEEOHDMUXEO-BQYQJAHWSA-N 0.000 claims abstract description 3
- MXXWOMGUGJBKIW-YPCIICBESA-N piperine Chemical compound C=1C=C2OCOC2=CC=1/C=C/C=C/C(=O)N1CCCCC1 MXXWOMGUGJBKIW-YPCIICBESA-N 0.000 claims abstract description 3
- WVWHRXVVAYXKDE-UHFFFAOYSA-N piperine Natural products O=C(C=CC=Cc1ccc2OCOc2c1)C3CCCCN3 WVWHRXVVAYXKDE-UHFFFAOYSA-N 0.000 claims abstract description 3
- 235000019100 piperine Nutrition 0.000 claims abstract description 3
- 229940075559 piperine Drugs 0.000 claims abstract description 3
- FPGPDEPMWUWLOV-UHFFFAOYSA-N polygodial Natural products CC1(C)CCCC2(C)C(C=O)C(=CC(O)C12)C=O FPGPDEPMWUWLOV-UHFFFAOYSA-N 0.000 claims abstract description 3
- 229940126586 small molecule drug Drugs 0.000 claims description 28
- 239000000243 solution Substances 0.000 claims description 24
- 239000003960 organic solvent Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 11
- 238000003756 stirring Methods 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 8
- 238000011282 treatment Methods 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 6
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 6
- 239000000568 immunological adjuvant Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 3
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 claims description 2
- 239000002202 Polyethylene glycol Substances 0.000 claims description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 2
- 229930003427 Vitamin E Natural products 0.000 claims description 2
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 2
- 230000001571 immunoadjuvant effect Effects 0.000 claims description 2
- 229960000502 poloxamer Drugs 0.000 claims description 2
- 229920001983 poloxamer Polymers 0.000 claims description 2
- 229920001223 polyethylene glycol Polymers 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 claims description 2
- 235000019165 vitamin E Nutrition 0.000 claims description 2
- 229940046009 vitamin E Drugs 0.000 claims description 2
- 239000011709 vitamin E Substances 0.000 claims description 2
- 229960001701 chloroform Drugs 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 4
- 238000004520 electroporation Methods 0.000 abstract description 4
- 238000009169 immunotherapy Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 21
- 239000002246 antineoplastic agent Substances 0.000 description 11
- 229940044683 chemotherapy drug Drugs 0.000 description 11
- 238000010586 diagram Methods 0.000 description 10
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 9
- 229910001424 calcium ion Inorganic materials 0.000 description 9
- 206010006187 Breast cancer Diseases 0.000 description 8
- 208000026310 Breast neoplasm Diseases 0.000 description 8
- 230000001737 promoting effect Effects 0.000 description 8
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 230000004041 dendritic cell maturation Effects 0.000 description 6
- 230000002147 killing effect Effects 0.000 description 6
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 6
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 6
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 6
- 239000011575 calcium Substances 0.000 description 5
- 230000001965 increasing effect Effects 0.000 description 5
- 210000001700 mitochondrial membrane Anatomy 0.000 description 5
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- 229910052791 calcium Inorganic materials 0.000 description 4
- 230000011748 cell maturation Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000001939 inductive effect Effects 0.000 description 4
- 230000035800 maturation Effects 0.000 description 4
- 230000028161 membrane depolarization Effects 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 210000001165 lymph node Anatomy 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- 108091006146 Channels Proteins 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 102000004082 Calreticulin Human genes 0.000 description 1
- 108090000549 Calreticulin Proteins 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 102000003563 TRPV Human genes 0.000 description 1
- 108060008564 TRPV Proteins 0.000 description 1
- 108010062740 TRPV Cation Channels Proteins 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- 102100029613 Transient receptor potential cation channel subfamily V member 1 Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 229940009456 adriamycin Drugs 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000008809 cell oxidative stress Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 108010010165 curculin Proteins 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical group O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000006882 induction of apoptosis Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000012798 spherical particle Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
Images
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种钙超载型小分子药物自组装纳米粒子及其应用,属于生物医药技术领域。所述纳米粒子由姜黄素、TRPV1激动剂和稳定剂制成;所述TRPV1激动剂选自辣椒素、树脂毒素、维生素D、胡椒碱、吴茱萸碱、花椒麻素、辣椒素酯、6‑姜烯酚、姜辣素、姜油酮、姜酮酚或水蓼二醛。本发明通过采用TRPV1激动剂和姜黄素联合制成的纳米粒子,以取代成本较高且繁琐的电穿孔技术,能够实现对于肿瘤的钙超载和免疫治疗的效果。
Description
技术领域
本发明属于生物医药技术领域,具体涉及一种钙超载型小分子药物自组装纳米粒子及其在制备免疫佐剂中的应用。
背景技术
恶性肿瘤是当前严重威胁人类生命健康的重大疾病之一。近年来,利用离子干扰疗法特别是钙离子进行肿瘤治疗逐渐成为研究人员关心的热点。钙离子是人体的一种常量元素,在人体内部的平衡稳态与细胞代谢和增殖密切相关。目前已有研究指出,钙离子Ca2+作为细胞信号传导的第二信使,参与了细胞凋亡信号等过程的传导。若体内钙离子平衡稳态被打破,钙离子充分进入胞质,便会导致细胞钙超载(Calcium Overload)。当前,学术界对于钙超载这一新型的诱导肿瘤细胞凋亡形式的关注逐年提高。
目前在临床研究上,主要通过钙电穿孔的方式诱导细胞钙超载进行治疗。钙电穿孔作为一种治疗恶性肿瘤的手段,其突出的优点是相比于传统的手术切除,其针对皮下转移瘤的治疗更加容易,无需进行手术即可基本消除肿瘤。但是其仍存在许多问题和缺点,其一,此种方法需要额外的电脉冲仪器,使用成本高;其二,此种方法治疗范围窄,目前在临床研究中仅适用于皮下转移瘤;其三,此方法首先需要在患处注射大量氯化钙,对于心脏功能不全者可能不适用。
2020年,诺贝尔奖聚焦于辣椒素受体(TRPV1),这种受体在受到药物刺激后能够打开,使得细胞间质液中高浓度的Ca2+进入细胞内部,因此可以用来作为钙超载的诱导手段。姜黄素(Curcumin)作为经典的抗癌药物,能够促进细胞内质网钙离子储库释放出钙并抑制钙的外排。因此,将二者结合,可以“内外兼修”的促进细胞内部钙离子水平的提高且不会增加心脏负担。目前未见有将TRPV1激动剂和姜黄素联合用来诱导钙超载的应用。
发明内容
本发明的目的是提供一种钙超载型小分子药物自组装纳米粒子,通过采用TRPV1激动剂和姜黄素联合的纳米粒子取代成本较高且繁琐的电穿孔技术,能够实现对于肿瘤的钙超载和免疫治疗的效果。
为了实现上述目的,本发明采用以下技术方案:
一种钙超载型小分子药物自组装纳米粒子,由姜黄素、TRPV1激动剂和稳定剂制成;
所述TRPV1激动剂选自辣椒素、树脂毒素、维生素D、胡椒碱、吴茱萸碱、花椒麻素、辣椒素酯、6-姜烯酚、姜辣素、姜油酮、姜酮酚或水蓼二醛;
所述稳定剂选自吐温80、维生素E聚乙二醇琥珀酸酯、聚氧乙烯吡咯烷酮、聚乙烯醇、DSPE-PEG2000或泊洛沙姆。
进一步地,所述TRPV1激动剂为辣椒素或树脂毒素。
进一步地,所述稳定剂为聚氧乙烯吡咯烷酮或DSPE-PEG2000。
上述钙超载型小分子药物自组装纳米粒子的制备方法,包括以下步骤:
步骤1,将姜黄素溶解于有机溶剂中,浓度为1~50 mg/mL,优选为10~20 mg/mL;
步骤2,将TRPV1激动剂溶解于有机溶剂中,浓度为1~50 mg/mL,优选为10~20 mg/mL;
步骤3,将稳定剂溶解于水中,浓度为1~100 % w/w,优选为1%~10% w/w;
步骤4,将步骤1的姜黄素溶液和步骤2的TRPV1激动剂溶液混合,然后加至步骤3的稳定剂溶液中混合;
步骤5,将步骤4得到的混合溶液加到水中,于10~80℃下100-1000 rpm搅拌0.1~4h后,停止搅拌,静置24 h除去有机溶剂,即得到钙超载型小分子自组装纳米粒子。
进一步地,所述有机溶剂为乙醇、二氯甲烷、三氯甲烷、二甲基亚砜、丙酮、乙腈或四氢呋喃。优选乙醇、四氢呋喃或丙酮。
进一步地,将步骤4得到的混合溶液加到水中,于10~50℃下200-800 rpm搅拌0.1~2 h后,停止搅拌,静置24 h除去有机溶剂。优选地,于20~30℃下200-400 rpm搅拌0.1~5h。
上述钙超载型小分子药物自组装纳米粒子在制备免疫佐剂中的应用。
上述钙超载型小分子药物自组装纳米粒子在制备肿瘤治疗药物中的应用。
本发明利用姜黄素在水中能够自组装形成纳米粒子以及其和TRPV1激动剂能够依赖疏水作用力形成纳米粒子的能力制备得到小分子药物自组装纳米粒子。该纳米粒子使得姜黄素和TRPV1激动剂发挥协同作用,促进细胞内部钙离子水平的提高且不会增加心脏负担。由于钙超载能够促进细胞免疫原死亡过程中的危险信号模式分子的释放(DAMPs),因此还可以将其应用拓展并作为免疫佐剂和化疗药物联用。
附图说明
图1为小分子药物自组装纳米粒子的外观和结构示意图。
图2为小分子药物自组装纳米粒子的粒径分布和外观图。
图3为小分子药物自组装纳米粒子对于4T1肿瘤细胞杀伤结果。
图4为小分子药物自组装纳米粒子促进树突状细胞成熟的结果。
图5为小分子药物自组装纳米粒子诱导线粒体膜电位去极化结果。
图6为小分子药物自组装纳米粒子和化疗药物联合后对于4T1乳腺癌细胞杀伤结果。
图7为小分子药物自组装纳米粒子促进4T1乳腺癌细胞表面CRT暴露结果。
图8为小分子药物自组装纳米粒子处理4T1乳腺癌细胞后促进树突状细胞成熟结果。
图9为小分子药物自组装纳米粒子促进小鼠淋巴结树突状细胞成熟结果。
图10为小分子药物自组装纳米粒子促进小鼠肿瘤CD8+ T细胞浸润结果。
图11为小分子药物自组装纳米粒子抑制4T1乳腺癌生长结果。
具体实施方式
下面结合附图和具体实施例对本发明作进一步详细说明,但不应理解为对本发明的限制。在不背离本发明精神和实质的情况下,对本发明方法、步骤或条件所作的修改或替换,均属于本发明的范围。实施例中未注明具体条件的实验方法及未说明配方的试剂均为按照本领域常规条件。
实施例1
步骤1,取2 mg和姜黄素和2 mg辣椒素分别加入装有0.1 mL四氢呋喃和0.1 mL无水乙醇的1.5 mL离心管中,分别得到姜黄素溶液和辣椒素溶液。
步骤2,取100 mg 聚氧乙烯吡咯烷酮加入到装有1 mL水的1.5 mL离心管中,得到10 %(w/w)浓度的聚氧乙烯吡咯烷酮溶液。
步骤3,将步骤(1)获得的姜黄素溶液和辣椒素溶液缓慢加入到步骤(2)获得的聚氧乙烯吡咯烷酮溶液中。
步骤4,将步骤(3)获得的混合溶液快速加入到装有20 mL水的烧瓶中,在26℃,400rpm搅拌0.5 h后静置12 h后除去有机溶剂。
步骤5,将步骤(4)获得的纳米粒子通过3000 kDa超滤管除去未包载的游离药物后得到钙超载型小分子自组装纳米粒子。
图1为小分子药物自组装纳米粒子的外观和示意图。由图(A)看出纳米粒子丁达尔效应明显,且无明显沉淀;图(B)为纳米粒子的示意图。
图2为小分子药物自组装纳米粒子的粒径分布(A)和透射电镜图(B)。由图可知制备得到的纳米粒子平均粒径Diameter 约为220 nm,多分散系数PdI为0.212±0.032的球形粒子。通过透射电镜可以观察到形成了符合上述描述的粒径均一的类球形结构。
图3为小分子药物自组装纳米粒子对于4T1肿瘤细胞杀伤示意图。通过MTT法评价了纳米粒子对于4T1细胞的细胞毒性。药物处理后加入噻唑蓝(浓度为0.5 mg/mL)4 h后加入二甲基亚砜(DMSO)后测定其在490 nm处的吸光度值。随后计算得到其IC50值约为24.14 μg/mL。
图4为小分子药物自组装纳米粒子促进树突状细胞成熟的流式图。通过流式细胞仪检测,经过不同药物处理后的树突状细胞表面标志物的表达情况。本实验利用CD80和CD86两种抗体作为评价DC细胞成熟的标志物。由图中可以看出,和对照组相比,游离的辣椒素和姜黄素相比促进DC细胞成熟的能力较弱,而所述纳米粒子能够有效促进DC细胞成熟,百分比约为85.9%,说明纳米粒子能够作为优良的免疫佐剂。
图5为小分子药物自组装纳米粒子诱导线粒体膜电位去极化示意图。通过流式细胞仪检测,经过不同药物处理后的4T1细胞线粒体膜电位的变化情况。钙超载会导致细胞氧化应激,进一步诱导线粒体膜电位去极化。利用JC-1探针可以检测膜电位的变化情况,当膜电位下降时,JC-1会以单体形式存在并在激光照射下发出绿色荧光,表现为FITC信号通道上升;反之细胞在正常状态下, JC-1以多聚体形式存在,在激光照射下发出红色荧光,表现为PE通道信号上升。从图中结果可以看出,纳米粒子处理24 h后细胞内具有最高的绿色荧光比例,说明促进线粒体膜电位去极化能力最强。
图6为小分子药物自组装纳米粒子和化疗药物联合后对于4T1乳腺癌细胞杀伤示意图。通过MTT法评价了和化疗药物阿霉素联合给药24 h后,不同浓度纳米粒子对于4T1肿瘤细胞的杀伤能力。由图可以看出,随着纳米粒子浓度的提高,联合杀伤能力显著增强。在20 μg/mL时,和对照组仅给予阿霉素相比,细胞毒性从82%降低至21%。
图7为小分子药物自组装纳米粒子促进4T1乳腺癌细胞表面CRT暴露示意图。CRT的暴露程度变化反映了细胞免疫原性死亡(ICD)的程度。通过流式细胞仪检测,经过不同药物处理24 h后4T1细胞表面钙网蛋白CRT暴露变化情况。定量结果可以看出,纳米粒子可以有效增强化疗药物诱导细胞暴露出最多的CRT,同时说明纳米粒子诱导的细胞死亡具有最强的免疫原性。
图8为小分子药物自组装纳米粒子处理4T1乳腺癌细胞后促进树突状细胞成熟示意图。在4T1细胞经过不同处理12h后,将其与DC细胞进行共孵育。共孵育后,DC成熟度的变化,反映了肿瘤死亡后引起免疫反应的程度。通过流式细胞仪检测DC细胞的成熟情况。由图中可以看出,纳米药物和化疗药物联合后能够有效促进DC细胞成熟,百分比为53.9%。
图9为小分子药物自组装纳米粒子促进小鼠淋巴结树突状细胞成熟示意图。由图中可以看出,纳米药物和化疗药物联合后能够有效促进小鼠体内淋巴结DC细胞的成熟,成熟度约为69.1%,说明纳米粒子能够作为优良的免疫佐剂。
图10为小分子药物自组装纳米粒子促进小鼠肿瘤CD8+ T细胞浸润示意图。CD8+ T细胞在肿瘤部位的提高说明小鼠对于肿瘤免疫能力的提高。由图中结果可以看出,纳米药物和化疗药物联合给药后,小鼠肿瘤部位CD8+ T细胞比例显著上升,相比对照上升3倍,说明纳米粒子能够提高化疗药物诱导的免疫效应,是有效的免疫佐剂。
图11为小分子药物自组装纳米粒子抑制4T1乳腺癌生长示意图。由图可以看出,纳米粒子和化疗药物联合给药后,肿瘤生长显著抑制,定量结果显示,和对照相比,肿瘤最终重量约为其1/10,和化疗药物阿霉素组相比,约为其1/4。说明纳米粒子能够有效增强化疗药物对于肿瘤的治疗效果。
实施例2
步骤1,取2 mg和姜黄素和2 mg树脂毒素分别加入装有0.1 mL四氢呋喃和0.1 mL无水乙醇的1.5 mL离心管中,分别得到姜黄素溶液和树脂毒素溶液。
步骤2,取100 mg 聚氧乙烯吡咯烷酮加入到装有1 mL水的1.5 mL离心管中,得到10 %(w/w)浓度的聚氧乙烯吡咯烷酮溶液。
步骤3,将步骤(1)获得的姜黄素溶液和树脂毒素溶液缓慢加入到步骤(2)获得的聚氧乙烯吡咯烷酮溶液中。
步骤4,将步骤(3)获得的混合溶液快速加入到装有20 mL水的烧瓶中,在26℃,400rpm搅拌0.5 h后静置12 h后除去有机溶剂。
步骤5,将步骤(4)获得的纳米粒子通过3000 kDa超滤管除去未包载的游离药物后得到钙超载型小分子自组装纳米粒子。
实施例3
步骤1,取2 mg和姜黄素和2 mg维生素D分别加入装有0.1 mL四氢呋喃和0.1 mL无水乙醇的1.5 mL离心管中,分别得到姜黄素溶液和维生素D溶液。
步骤2,取100 mg 聚氧乙烯吡咯烷酮加入到装有1 mL水的1.5 mL离心管中,得到10 %(w/w)浓度的聚氧乙烯吡咯烷酮溶液。
步骤3,将步骤(1)获得的姜黄素溶液和维生素D溶液缓慢加入到步骤(2)获得的聚氧乙烯吡咯烷酮溶液中。
步骤4,将步骤(3)获得的混合溶液快速加入到装有20 mL水的烧瓶中,在26℃,400rpm搅拌0.5 h后静置12 h后除去有机溶剂。
步骤5,将步骤(4)获得的纳米粒子通过3000 kDa超滤管除去未包载的游离药物后得到钙超载型小分子自组装纳米粒子。
Claims (8)
1.一种钙超载型小分子药物自组装纳米粒子,其特征在于:由姜黄素、TRPV1激动剂和稳定剂制成;
所述TRPV1激动剂选自辣椒素、树脂毒素、维生素D、胡椒碱、吴茱萸碱、花椒麻素、辣椒素酯、6-姜烯酚、姜辣素、姜油酮、姜酮酚或水蓼二醛;
所述稳定剂选自吐温80、维生素E聚乙二醇琥珀酸酯、聚氧乙烯吡咯烷酮、聚乙烯醇、DSPE-PEG2000或泊洛沙姆。
2.根据权利要求1所述的钙超载型小分子药物自组装纳米粒子,其特征在于:所述TRPV1激动剂为辣椒素或树脂毒素。
3.根据权利要求1所述的钙超载型小分子药物自组装纳米粒子,其特征在于:所述稳定剂为聚氧乙烯吡咯烷酮或DSPE-PEG2000。
4.权利要求1所述的钙超载型小分子药物自组装纳米粒子的制备方法,其特征在于:包括以下步骤:
步骤1,将姜黄素溶解于有机溶剂中,浓度为1~50 mg/mL;
步骤2,将TRPV1激动剂溶解于有机溶剂中,浓度为1~50 mg/mL;
步骤3,将稳定剂溶解于水中,浓度为1~100 % w/w;
步骤4,将步骤1的姜黄素溶液和步骤2的TRPV1激动剂溶液混合,然后加至步骤3的稳定剂溶液中混合;
步骤5,将步骤4得到的混合溶液加到水中,于10~80℃下100-1000 rpm搅拌0.1~4 h后,停止搅拌,静置24 h除去有机溶剂,即得到钙超载型小分子自组装纳米粒子。
5.根据权利要求4所述的制备方法,其特征在于:所述有机溶剂为乙醇、二氯甲烷、三氯甲烷、二甲基亚砜、丙酮、乙腈或四氢呋喃。
6.根据权利要求4所述的制备方法,其特征在于:将步骤4得到的混合溶液加到水中,于10~50℃下200-800 rpm搅拌0.1~2 h后,停止搅拌,静置24 h除去有机溶剂。
7.权利要求1所述的钙超载型小分子药物自组装纳米粒子在制备免疫佐剂中的应用。
8.权利要求1所述的钙超载型小分子药物自组装纳米粒子在制备肿瘤治疗药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210114493.6A CN114306609A (zh) | 2022-01-30 | 2022-01-30 | 一种钙超载型小分子药物自组装纳米粒子及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210114493.6A CN114306609A (zh) | 2022-01-30 | 2022-01-30 | 一种钙超载型小分子药物自组装纳米粒子及其应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114306609A true CN114306609A (zh) | 2022-04-12 |
Family
ID=81031323
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210114493.6A Pending CN114306609A (zh) | 2022-01-30 | 2022-01-30 | 一种钙超载型小分子药物自组装纳米粒子及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114306609A (zh) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103784421A (zh) * | 2014-02-27 | 2014-05-14 | 哈尔滨医科大学 | 载姜黄素和胡椒碱的固体脂质纳米粒及其制备方法 |
-
2022
- 2022-01-30 CN CN202210114493.6A patent/CN114306609A/zh active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103784421A (zh) * | 2014-02-27 | 2014-05-14 | 哈尔滨医科大学 | 载姜黄素和胡椒碱的固体脂质纳米粒及其制备方法 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Hu et al. | The potentiated checkpoint blockade immunotherapy by ROS-responsive nanocarrier-mediated cascade chemo-photodynamic therapy | |
Chen et al. | PLGA-PEG-PLGA triblock copolymeric micelles as oral drug delivery system: In vitro drug release and in vivo pharmacokinetics assessment | |
Gao et al. | Glutathione-responsive nanoparticles based on a sodium alginate derivative for selective release of doxorubicin in tumor cells | |
Wei et al. | Multifunctional polymeric micelle-based chemo-immunotherapy with immune checkpoint blockade for efficient treatment of orthotopic and metastatic breast cancer | |
Guo et al. | Novel Chinese angelica polysaccharide biomimetic nanomedicine to curcumin delivery for hepatocellular carcinoma treatment and immunomodulatory effect | |
Sun et al. | Enhanced antitumor efficacy of vitamin E TPGS-emulsified PLGA nanoparticles for delivery of paclitaxel | |
Zhang et al. | Redox-and light-responsive alginate nanoparticles as effective drug carriers for combinational anticancer therapy | |
CN107669632B (zh) | 药物载体、胶束、药物制剂、及其制备方法和用途 | |
EP2515946A2 (en) | Nanoconjugates and nanoconjugate formulations | |
JP7164205B2 (ja) | キナ酸修飾ナノ粒子およびその使用 | |
Alagheband et al. | Design and fabrication of a dual-drug loaded nano-platform for synergistic anticancer and cytotoxicity effects on the expression of leptin in lung cancer treatment | |
Shang et al. | Enhancing cancer chemo-immunotherapy by biomimetic nanogel with tumor targeting capacity and rapid drug-releasing in tumor microenvironment | |
Zong et al. | Self-amplified chain-shattering cinnamaldehyde-based poly (thioacetal) boosts cancer chemo-immunotherapy | |
Zu et al. | Docetaxel-loaded bovine serum albumin nanoparticles conjugated docosahexaenoic acid for inhibiting lung cancer metastasis to bone | |
Liu et al. | Thermosensitive selenium hydrogel boosts antitumor immune response for hepatocellular carcinoma chemoradiotherapy | |
JP2018516256A (ja) | 治療用ワクチンとしての使用のためのナノ粒子 | |
Liang et al. | Crizotinib prodrug micelles co-delivered doxorubicin for synergistic immunogenic cell death induction on breast cancer chemo-immunotherapy | |
Jeswani et al. | Design of vincristine sulfate loaded poloxamer in situ nanogel: formulation and in vitro evaluation | |
WO2022228469A1 (zh) | 聚合物囊泡纳米sting激动剂及其制备方法与应用 | |
Sun et al. | Supramolecular nanomedicine for selective cancer therapy via sequential responsiveness to reactive oxygen species and glutathione | |
CN107007550B (zh) | 一种氧化还原响应性两亲性共聚物及其制备方法和应用 | |
CN113456614A (zh) | 一种基于plga的粒径可变型抗肿瘤仿生纳米制剂及其制备方法和应用 | |
Ma et al. | Biodegradable methoxy poly (ethylene glycol)-poly (lactide) nanoparticles for controlled delivery of dacarbazine: preparation, characterization and anticancer activity evaluation | |
Wu et al. | Development and characterization of DEC-205 receptor targeted potentilla anserina l polysaccharide PLGA nanoparticles as an antigen delivery system to enhance in vitro and in vivo immune responses in mice | |
Huang et al. | Black phosphorus assisted polyionic micelles with efficient PTX loading for remotely controlled release and synergistic treatment of drug-resistant tumors |
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 | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220412 |