US20210330809A1 - Lipid-coated particles for treating viral infections - Google Patents
Lipid-coated particles for treating viral infections Download PDFInfo
- Publication number
- US20210330809A1 US20210330809A1 US17/306,452 US202117306452A US2021330809A1 US 20210330809 A1 US20210330809 A1 US 20210330809A1 US 202117306452 A US202117306452 A US 202117306452A US 2021330809 A1 US2021330809 A1 US 2021330809A1
- Authority
- US
- United States
- Prior art keywords
- optionally substituted
- lipid
- mol
- msns
- group
- 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.)
- Abandoned
Links
- 150000002632 lipids Chemical class 0.000 title claims abstract description 155
- 239000002245 particle Substances 0.000 title claims abstract description 142
- 208000036142 Viral infection Diseases 0.000 title abstract description 17
- 230000009385 viral infection Effects 0.000 title abstract description 16
- 150000001875 compounds Chemical class 0.000 claims abstract description 111
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 76
- 238000000034 method Methods 0.000 claims abstract description 52
- -1 nitro, amino Chemical group 0.000 claims description 88
- 239000002953 phosphate buffered saline Substances 0.000 claims description 63
- 241000710959 Venezuelan equine encephalitis virus Species 0.000 claims description 47
- 239000011148 porous material Substances 0.000 claims description 42
- 150000003839 salts Chemical class 0.000 claims description 36
- 230000003612 virological effect Effects 0.000 claims description 35
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 31
- 238000000338 in vitro Methods 0.000 claims description 26
- 125000000623 heterocyclic group Chemical group 0.000 claims description 23
- 210000004556 brain Anatomy 0.000 claims description 18
- 125000000547 substituted alkyl group Chemical group 0.000 claims description 17
- 235000012000 cholesterol Nutrition 0.000 claims description 15
- 239000007864 aqueous solution Substances 0.000 claims description 14
- 125000005010 perfluoroalkyl group Chemical group 0.000 claims description 13
- 125000002877 alkyl aryl group Chemical group 0.000 claims description 11
- 125000001188 haloalkyl group Chemical group 0.000 claims description 11
- 230000002209 hydrophobic effect Effects 0.000 claims description 11
- 125000002768 hydroxyalkyl group Chemical group 0.000 claims description 11
- 238000000576 coating method Methods 0.000 claims description 10
- 125000005346 substituted cycloalkyl group Chemical group 0.000 claims description 10
- 125000000852 azido group Chemical group *N=[N+]=[N-] 0.000 claims description 9
- 238000007912 intraperitoneal administration Methods 0.000 claims description 8
- 239000011248 coating agent Substances 0.000 claims description 7
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 7
- 125000003107 substituted aryl group Chemical group 0.000 claims description 7
- 238000000423 cell based assay Methods 0.000 claims description 6
- 238000001990 intravenous administration Methods 0.000 claims description 5
- 241000710929 Alphavirus Species 0.000 claims description 4
- 206010028980 Neoplasm Diseases 0.000 claims description 4
- 201000011510 cancer Diseases 0.000 claims description 4
- 238000007918 intramuscular administration Methods 0.000 claims description 4
- 208000007887 Alphavirus Infections Diseases 0.000 claims description 3
- 238000010254 subcutaneous injection Methods 0.000 claims description 2
- 239000007929 subcutaneous injection Substances 0.000 claims description 2
- 125000001475 halogen functional group Chemical group 0.000 claims 4
- KSXTUUUQYQYKCR-LQDDAWAPSA-M 2,3-bis[[(z)-octadec-9-enoyl]oxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC KSXTUUUQYQYKCR-LQDDAWAPSA-M 0.000 claims 1
- 230000001093 anti-cancer Effects 0.000 claims 1
- 208000015181 infectious disease Diseases 0.000 abstract description 35
- 239000000969 carrier Substances 0.000 abstract description 5
- 206010014612 Encephalitis viral Diseases 0.000 abstract description 3
- 201000002498 viral encephalitis Diseases 0.000 abstract description 3
- DCAMTBFFNOHDAW-UHFFFAOYSA-N 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-5-nitro-n-phenylbenzamide Chemical compound C1N(C)CCN(C)C1=NC1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=CC=C1 DCAMTBFFNOHDAW-UHFFFAOYSA-N 0.000 description 146
- 210000004027 cell Anatomy 0.000 description 98
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 67
- 125000003118 aryl group Chemical group 0.000 description 50
- 125000000217 alkyl group Chemical group 0.000 description 46
- 239000003814 drug Substances 0.000 description 45
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 42
- 239000000203 mixture Substances 0.000 description 42
- 108090000765 processed proteins & peptides Proteins 0.000 description 40
- 229940079593 drug Drugs 0.000 description 38
- 241001465754 Metazoa Species 0.000 description 35
- 239000000232 Lipid Bilayer Substances 0.000 description 34
- 241000700605 Viruses Species 0.000 description 33
- 239000003112 inhibitor Substances 0.000 description 32
- 239000002105 nanoparticle Substances 0.000 description 30
- 239000000243 solution Substances 0.000 description 30
- 241000699670 Mus sp. Species 0.000 description 29
- 230000008685 targeting Effects 0.000 description 29
- 239000006228 supernatant Substances 0.000 description 26
- 239000003446 ligand Substances 0.000 description 25
- 229920001223 polyethylene glycol Polymers 0.000 description 25
- 239000012634 fragment Substances 0.000 description 22
- 239000002502 liposome Substances 0.000 description 22
- 238000011068 loading method Methods 0.000 description 22
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 20
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 20
- 238000001727 in vivo Methods 0.000 description 20
- 239000000377 silicon dioxide Substances 0.000 description 20
- 125000002947 alkylene group Chemical group 0.000 description 19
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 description 17
- 239000003795 chemical substances by application Substances 0.000 description 17
- 238000009472 formulation Methods 0.000 description 17
- 108090000623 proteins and genes Proteins 0.000 description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 16
- 125000005843 halogen group Chemical group 0.000 description 16
- 230000005764 inhibitory process Effects 0.000 description 16
- 102000004196 processed proteins & peptides Human genes 0.000 description 16
- 230000012202 endocytosis Effects 0.000 description 15
- 238000005516 engineering process Methods 0.000 description 15
- 235000018102 proteins Nutrition 0.000 description 15
- 102000004169 proteins and genes Human genes 0.000 description 15
- 210000001519 tissue Anatomy 0.000 description 15
- 238000011282 treatment Methods 0.000 description 15
- 230000001413 cellular effect Effects 0.000 description 14
- 238000009826 distribution Methods 0.000 description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 13
- 230000006395 clathrin-mediated endocytosis Effects 0.000 description 13
- 229910052757 nitrogen Inorganic materials 0.000 description 13
- 150000003384 small molecules Chemical class 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 12
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 12
- 244000052769 pathogen Species 0.000 description 12
- 239000002202 Polyethylene glycol Substances 0.000 description 11
- 230000015572 biosynthetic process Effects 0.000 description 11
- 201000010099 disease Diseases 0.000 description 11
- SYNDQCRDGGCQRZ-VXLYETTFSA-N dynasore Chemical compound C1=C(O)C(O)=CC=C1\C=N\NC(=O)C1=CC2=CC=CC=C2C=C1O SYNDQCRDGGCQRZ-VXLYETTFSA-N 0.000 description 11
- 210000003734 kidney Anatomy 0.000 description 11
- 210000004185 liver Anatomy 0.000 description 11
- 210000000952 spleen Anatomy 0.000 description 11
- 238000005119 centrifugation Methods 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 230000000670 limiting effect Effects 0.000 description 10
- 230000009467 reduction Effects 0.000 description 10
- 230000002829 reductive effect Effects 0.000 description 10
- 231100000419 toxicity Toxicity 0.000 description 10
- 230000001988 toxicity Effects 0.000 description 10
- KWVJHCQQUFDPLU-YEUCEMRASA-N 2,3-bis[[(z)-octadec-9-enoyl]oxy]propyl-trimethylazanium Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC KWVJHCQQUFDPLU-YEUCEMRASA-N 0.000 description 9
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 9
- SUHOOTKUPISOBE-UHFFFAOYSA-N O-phosphoethanolamine Chemical compound NCCOP(O)(O)=O SUHOOTKUPISOBE-UHFFFAOYSA-N 0.000 description 9
- 239000003443 antiviral agent Substances 0.000 description 9
- 230000004087 circulation Effects 0.000 description 9
- 208000035475 disorder Diseases 0.000 description 9
- 229910052739 hydrogen Inorganic materials 0.000 description 9
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 9
- 239000000546 pharmaceutical excipient Substances 0.000 description 9
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 9
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 8
- 241000713124 Rift Valley fever virus Species 0.000 description 8
- 241000711975 Vesicular stomatitis virus Species 0.000 description 8
- XDHNQDDQEHDUTM-UHFFFAOYSA-N bafliomycin A1 Natural products COC1C=CC=C(C)CC(C)C(O)C(C)C=C(C)C=C(OC)C(=O)OC1C(C)C(O)C(C)C1(O)OC(C(C)C)C(C)C(O)C1 XDHNQDDQEHDUTM-UHFFFAOYSA-N 0.000 description 8
- 230000008901 benefit Effects 0.000 description 8
- 150000001720 carbohydrates Chemical class 0.000 description 8
- 125000002091 cationic group Chemical group 0.000 description 8
- 231100000673 dose–response relationship Toxicity 0.000 description 8
- 238000012377 drug delivery Methods 0.000 description 8
- 239000000975 dye Substances 0.000 description 8
- 238000002296 dynamic light scattering Methods 0.000 description 8
- 239000001257 hydrogen Substances 0.000 description 8
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 8
- 230000001717 pathogenic effect Effects 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- XDHNQDDQEHDUTM-XJKSCTEHSA-N (3z,5e,7r,8s,9r,11e,13e,15s,16r)-16-[(2s,3r,4s)-4-[(2r,4r,5s,6r)-2,4-dihydroxy-5-methyl-6-propan-2-yloxan-2-yl]-3-hydroxypentan-2-yl]-8-hydroxy-3,15-dimethoxy-5,7,9,11-tetramethyl-1-oxacyclohexadeca-3,5,11,13-tetraen-2-one Chemical compound CO[C@H]1\C=C\C=C(C)\C[C@@H](C)[C@H](O)[C@H](C)\C=C(/C)\C=C(OC)\C(=O)O[C@@H]1[C@@H](C)[C@@H](O)[C@H](C)[C@]1(O)O[C@H](C(C)C)[C@@H](C)[C@H](O)C1 XDHNQDDQEHDUTM-XJKSCTEHSA-N 0.000 description 7
- LVNGJLRDBYCPGB-LDLOPFEMSA-N (R)-1,2-distearoylphosphatidylethanolamine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[NH3+])OC(=O)CCCCCCCCCCCCCCCCC LVNGJLRDBYCPGB-LDLOPFEMSA-N 0.000 description 7
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 7
- PHEDXBVPIONUQT-UHFFFAOYSA-N Cocarcinogen A1 Natural products CCCCCCCCCCCCCC(=O)OC1C(C)C2(O)C3C=C(C)C(=O)C3(O)CC(CO)=CC2C2C1(OC(C)=O)C2(C)C PHEDXBVPIONUQT-UHFFFAOYSA-N 0.000 description 7
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 7
- 241000282412 Homo Species 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 125000000753 cycloalkyl group Chemical group 0.000 description 7
- 230000001419 dependent effect Effects 0.000 description 7
- 238000000684 flow cytometry Methods 0.000 description 7
- 230000007246 mechanism Effects 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 7
- 230000007935 neutral effect Effects 0.000 description 7
- PHEDXBVPIONUQT-RGYGYFBISA-N phorbol 13-acetate 12-myristate Chemical compound C([C@]1(O)C(=O)C(C)=C[C@H]1[C@@]1(O)[C@H](C)[C@H]2OC(=O)CCCCCCCCCCCCC)C(CO)=C[C@H]1[C@H]1[C@]2(OC(C)=O)C1(C)C PHEDXBVPIONUQT-RGYGYFBISA-N 0.000 description 7
- 238000000527 sonication Methods 0.000 description 7
- 239000003981 vehicle Substances 0.000 description 7
- QDLHCMPXEPAAMD-UHFFFAOYSA-N wortmannin Natural products COCC1OC(=O)C2=COC(C3=O)=C2C1(C)C1=C3C2CCC(=O)C2(C)CC1OC(C)=O QDLHCMPXEPAAMD-UHFFFAOYSA-N 0.000 description 7
- NRJAVPSFFCBXDT-HUESYALOSA-N 1,2-distearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCCCC NRJAVPSFFCBXDT-HUESYALOSA-N 0.000 description 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 239000000654 additive Substances 0.000 description 6
- 230000000996 additive effect Effects 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 235000014633 carbohydrates Nutrition 0.000 description 6
- 125000004093 cyano group Chemical group *C#N 0.000 description 6
- 230000000021 endosomolytic effect Effects 0.000 description 6
- 230000000799 fusogenic effect Effects 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 230000003993 interaction Effects 0.000 description 6
- 210000004072 lung Anatomy 0.000 description 6
- 239000003550 marker Substances 0.000 description 6
- 229910052751 metal Inorganic materials 0.000 description 6
- 239000002184 metal Substances 0.000 description 6
- 239000002086 nanomaterial Substances 0.000 description 6
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 6
- TXLHNFOLHRXMAU-UHFFFAOYSA-N 2-(4-benzylphenoxy)-n,n-diethylethanamine;hydron;chloride Chemical compound Cl.C1=CC(OCCN(CC)CC)=CC=C1CC1=CC=CC=C1 TXLHNFOLHRXMAU-UHFFFAOYSA-N 0.000 description 5
- RFAXLXKIAKIUDT-UHFFFAOYSA-N IPA-3 Chemical compound C1=CC=C2C(SSC3=C4C=CC=CC4=CC=C3O)=C(O)C=CC2=C1 RFAXLXKIAKIUDT-UHFFFAOYSA-N 0.000 description 5
- 229930182558 Sterol Natural products 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 150000001413 amino acids Chemical group 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
- 238000012512 characterization method Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 229910052731 fluorine Inorganic materials 0.000 description 5
- 125000001072 heteroaryl group Chemical group 0.000 description 5
- 230000003834 intracellular effect Effects 0.000 description 5
- 239000011859 microparticle Substances 0.000 description 5
- 238000000386 microscopy Methods 0.000 description 5
- 125000004433 nitrogen atom Chemical group N* 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 210000004940 nucleus Anatomy 0.000 description 5
- 230000037361 pathway Effects 0.000 description 5
- 125000006239 protecting group Chemical group 0.000 description 5
- 150000003432 sterols Chemical class 0.000 description 5
- 235000003702 sterols Nutrition 0.000 description 5
- 125000001424 substituent group Chemical group 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 125000006552 (C3-C8) cycloalkyl group Chemical group 0.000 description 4
- KILNVBDSWZSGLL-KXQOOQHDSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCC KILNVBDSWZSGLL-KXQOOQHDSA-N 0.000 description 4
- SNKAWJBJQDLSFF-NVKMUCNASA-N 1,2-dioleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC SNKAWJBJQDLSFF-NVKMUCNASA-N 0.000 description 4
- ATHVAWFAEPLPPQ-VRDBWYNSSA-N 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC ATHVAWFAEPLPPQ-VRDBWYNSSA-N 0.000 description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 4
- 206010014611 Encephalitis venezuelan equine Diseases 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- QUSNBJAOOMFDIB-UHFFFAOYSA-N Ethylamine Chemical compound CCN QUSNBJAOOMFDIB-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 4
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 description 4
- 102000029749 Microtubule Human genes 0.000 description 4
- 108091022875 Microtubule Proteins 0.000 description 4
- 208000012902 Nervous system disease Diseases 0.000 description 4
- 239000012124 Opti-MEM Substances 0.000 description 4
- 229930182555 Penicillin Natural products 0.000 description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 4
- KPKZJLCSROULON-QKGLWVMZSA-N Phalloidin Chemical compound N1C(=O)[C@@H]([C@@H](O)C)NC(=O)[C@H](C)NC(=O)[C@H](C[C@@](C)(O)CO)NC(=O)[C@H](C2)NC(=O)[C@H](C)NC(=O)[C@@H]3C[C@H](O)CN3C(=O)[C@@H]1CSC1=C2C2=CC=CC=C2N1 KPKZJLCSROULON-QKGLWVMZSA-N 0.000 description 4
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 4
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000027455 binding Effects 0.000 description 4
- GZUXJHMPEANEGY-UHFFFAOYSA-N bromomethane Chemical compound BrC GZUXJHMPEANEGY-UHFFFAOYSA-N 0.000 description 4
- 150000001721 carbon Chemical group 0.000 description 4
- 230000027448 caveolin-mediated endocytosis Effects 0.000 description 4
- GCFBRXLSHGKWDP-XCGNWRKASA-N cefoperazone Chemical compound O=C1C(=O)N(CC)CCN1C(=O)N[C@H](C=1C=CC(O)=CC=1)C(=O)N[C@@H]1C(=O)N2C(C(O)=O)=C(CSC=3N(N=NN=3)C)CS[C@@H]21 GCFBRXLSHGKWDP-XCGNWRKASA-N 0.000 description 4
- 230000001186 cumulative effect Effects 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 4
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 4
- 229940043264 dodecyl sulfate Drugs 0.000 description 4
- 238000000635 electron micrograph Methods 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 125000001153 fluoro group Chemical group F* 0.000 description 4
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 4
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- 230000034701 macropinocytosis Effects 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 210000004688 microtubule Anatomy 0.000 description 4
- 231100000252 nontoxic Toxicity 0.000 description 4
- 230000003000 nontoxic effect Effects 0.000 description 4
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 229940049954 penicillin Drugs 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 239000001294 propane Substances 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 239000004055 small Interfering RNA Substances 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 229910052717 sulfur Inorganic materials 0.000 description 4
- 239000011593 sulfur Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 229940124597 therapeutic agent Drugs 0.000 description 4
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 4
- 125000004400 (C1-C12) alkyl group Chemical group 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- FVXDQWZBHIXIEJ-LNDKUQBDSA-N 1,2-di-[(9Z,12Z)-octadecadienoyl]-sn-glycero-3-phosphocholine Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC FVXDQWZBHIXIEJ-LNDKUQBDSA-N 0.000 description 3
- UEPCKCAOFFAEMS-UHFFFAOYSA-N 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-n-(4-methoxyphenyl)-5-nitrobenzamide Chemical compound C1=CC(OC)=CC=C1NC(=O)C1=CC([N+]([O-])=O)=CC=C1N=C1N(C)CCN(C)C1 UEPCKCAOFFAEMS-UHFFFAOYSA-N 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 3
- 108010085238 Actins Proteins 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- BDCFUHIWJODVNG-UHFFFAOYSA-N Desmosterol Natural products C1C=C2CC(O)C=CC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 BDCFUHIWJODVNG-UHFFFAOYSA-N 0.000 description 3
- GZDFHIJNHHMENY-UHFFFAOYSA-N Dimethyl dicarbonate Chemical compound COC(=O)OC(=O)OC GZDFHIJNHHMENY-UHFFFAOYSA-N 0.000 description 3
- 241000283073 Equus caballus Species 0.000 description 3
- 241000710842 Japanese encephalitis virus Species 0.000 description 3
- 241001631646 Papillomaviridae Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 0 [1*]N([2*])C(=O)C1=CC([6*])=CC([3*])=C1/N=C1/N([1*])C([4*])([5*])C([4*])([5*])N([1*])C1([4*])[5*].[1*]N1C([4*])([5*])C([4*])([5*])N(C([7*])([8*])C2=NC3=C([3*])C=C([6*])C=C3C(=O)N2[2*])C([4*])([5*])C1([4*])[5*].[2*]C1=NC2=C([3*])C=C([6*])C=C2C(=O)N1[2*] Chemical compound [1*]N([2*])C(=O)C1=CC([6*])=CC([3*])=C1/N=C1/N([1*])C([4*])([5*])C([4*])([5*])N([1*])C1([4*])[5*].[1*]N1C([4*])([5*])C([4*])([5*])N(C([7*])([8*])C2=NC3=C([3*])C=C([6*])C=C3C(=O)N2[2*])C([4*])([5*])C1([4*])[5*].[2*]C1=NC2=C([3*])C=C([6*])C=C2C(=O)N1[2*] 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 125000002252 acyl group Chemical group 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 125000000129 anionic group Chemical group 0.000 description 3
- 229940121357 antivirals Drugs 0.000 description 3
- 239000003125 aqueous solvent Substances 0.000 description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 3
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 description 3
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 210000004323 caveolae Anatomy 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000013270 controlled release Methods 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 210000004443 dendritic cell Anatomy 0.000 description 3
- AVSXSVCZWQODGV-DPAQBDIFSA-N desmosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@@H](CCC=C(C)C)C)[C@@]1(C)CC2 AVSXSVCZWQODGV-DPAQBDIFSA-N 0.000 description 3
- 206010014599 encephalitis Diseases 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 210000003494 hepatocyte Anatomy 0.000 description 3
- 125000005842 heteroatom Chemical group 0.000 description 3
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 3
- 150000002431 hydrogen Chemical group 0.000 description 3
- 230000002458 infectious effect Effects 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 238000002429 nitrogen sorption measurement Methods 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 230000030648 nucleus localization Effects 0.000 description 3
- HGASFNYMVGEKTF-UHFFFAOYSA-N octan-1-ol;hydrate Chemical compound O.CCCCCCCCO HGASFNYMVGEKTF-UHFFFAOYSA-N 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 238000005192 partition Methods 0.000 description 3
- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 3
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 3
- 239000002096 quantum dot Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000001878 scanning electron micrograph Methods 0.000 description 3
- 238000004626 scanning electron microscopy Methods 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 229960005322 streptomycin Drugs 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 238000004627 transmission electron microscopy Methods 0.000 description 3
- 210000003501 vero cell Anatomy 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- LSPHULWDVZXLIL-UHFFFAOYSA-N (+/-)-Camphoric acid Chemical compound CC1(C)C(C(O)=O)CCC1(C)C(O)=O LSPHULWDVZXLIL-UHFFFAOYSA-N 0.000 description 2
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 2
- QYIXCDOBOSTCEI-QCYZZNICSA-N (5alpha)-cholestan-3beta-ol Chemical compound C([C@@H]1CC2)[C@@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)CCCC(C)C)[C@@]2(C)CC1 QYIXCDOBOSTCEI-QCYZZNICSA-N 0.000 description 2
- 125000004191 (C1-C6) alkoxy group Chemical group 0.000 description 2
- 125000004738 (C1-C6) alkyl sulfinyl group Chemical group 0.000 description 2
- 125000004739 (C1-C6) alkylsulfonyl group Chemical group 0.000 description 2
- 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 description 2
- WLXGQMVCYPUOLM-UHFFFAOYSA-N 1-hydroxyethanesulfonic acid Chemical compound CC(O)S(O)(=O)=O WLXGQMVCYPUOLM-UHFFFAOYSA-N 0.000 description 2
- AVRPFRMDMNDIDH-UHFFFAOYSA-N 1h-quinazolin-2-one Chemical class C1=CC=CC2=NC(O)=NC=C21 AVRPFRMDMNDIDH-UHFFFAOYSA-N 0.000 description 2
- YQTCQNIPQMJNTI-UHFFFAOYSA-N 2,2-dimethylpropan-1-one Chemical group CC(C)(C)[C]=O YQTCQNIPQMJNTI-UHFFFAOYSA-N 0.000 description 2
- FGYOIIHLPCHRAK-UHFFFAOYSA-N 2-(furan-2-yl)-4-[(2-methyl-6,8-dihydro-5h-pyrido[3,4-d]pyrimidin-7-yl)methyl]-1,3-thiazole Chemical compound C1C2=NC(C)=NC=C2CCN1CC(N=1)=CSC=1C1=CC=CO1 FGYOIIHLPCHRAK-UHFFFAOYSA-N 0.000 description 2
- KKZUIWZVXFPMAU-UHFFFAOYSA-N 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-n-(2-fluorophenyl)-5-nitrobenzamide Chemical compound C1N(C)CCN(C)C1=NC1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=CC=C1F KKZUIWZVXFPMAU-UHFFFAOYSA-N 0.000 description 2
- CFFXRIUELQDINN-UHFFFAOYSA-N 2-[(4-ethylpiperazin-1-yl)methyl]-3-(2-fluorophenyl)-6-nitroquinazolin-4-one Chemical compound C1CN(CC)CCN1CC1=NC2=CC=C([N+]([O-])=O)C=C2C(=O)N1C1=CC=CC=C1F CFFXRIUELQDINN-UHFFFAOYSA-N 0.000 description 2
- GRWKNBPOGBTZMN-UHFFFAOYSA-N 2-benzyl-3-phenylpropane-1,2-diamine Chemical compound C=1C=CC=CC=1CC(N)(CN)CC1=CC=CC=C1 GRWKNBPOGBTZMN-UHFFFAOYSA-N 0.000 description 2
- LBLYYCQCTBFVLH-UHFFFAOYSA-M 2-methylbenzenesulfonate Chemical compound CC1=CC=CC=C1S([O-])(=O)=O LBLYYCQCTBFVLH-UHFFFAOYSA-M 0.000 description 2
- 229940080296 2-naphthalenesulfonate Drugs 0.000 description 2
- FEWJPZIEWOKRBE-UHFFFAOYSA-M 3-carboxy-2,3-dihydroxypropanoate Chemical compound OC(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-M 0.000 description 2
- ALKYHXVLJMQRLQ-UHFFFAOYSA-M 3-carboxynaphthalen-2-olate Chemical compound C1=CC=C2C=C(C([O-])=O)C(O)=CC2=C1 ALKYHXVLJMQRLQ-UHFFFAOYSA-M 0.000 description 2
- ZRPLANDPDWYOMZ-UHFFFAOYSA-N 3-cyclopentylpropionic acid Chemical compound OC(=O)CCC1CCCC1 ZRPLANDPDWYOMZ-UHFFFAOYSA-N 0.000 description 2
- XMIIGOLPHOKFCH-UHFFFAOYSA-M 3-phenylpropionate Chemical compound [O-]C(=O)CCC1=CC=CC=C1 XMIIGOLPHOKFCH-UHFFFAOYSA-M 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- GTUCHTUUIMKCLD-UHFFFAOYSA-N 4-(6,8-dihydro-5h-pyrido[3,4-d]pyrimidin-7-ylmethyl)-2-(5-methylfuran-2-yl)-1,3-thiazole Chemical compound O1C(C)=CC=C1C1=NC(CN2CC3=NC=NC=C3CC2)=CS1 GTUCHTUUIMKCLD-UHFFFAOYSA-N 0.000 description 2
- HPNYOSMNUIUJBY-UHFFFAOYSA-N 5-cyano-2-[(1,4-dimethylpiperazin-2-ylidene)amino]-n-phenylbenzamide Chemical compound C1N(C)CCN(C)C1=NC1=CC=C(C#N)C=C1C(=O)NC1=CC=CC=C1 HPNYOSMNUIUJBY-UHFFFAOYSA-N 0.000 description 2
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- 108091023037 Aptamer Proteins 0.000 description 2
- 238000012935 Averaging Methods 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 2
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 2
- 241001136175 Burkholderia pseudomallei Species 0.000 description 2
- 239000004322 Butylated hydroxytoluene Substances 0.000 description 2
- NLZUEZXRPGMBCV-UHFFFAOYSA-N Butylhydroxytoluene Chemical compound CC1=CC(C(C)(C)C)=C(O)C(C(C)(C)C)=C1 NLZUEZXRPGMBCV-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 125000000882 C2-C6 alkenyl group Chemical group 0.000 description 2
- 125000003601 C2-C6 alkynyl group Chemical group 0.000 description 2
- 108091033409 CRISPR Proteins 0.000 description 2
- 238000010354 CRISPR gene editing Methods 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 description 2
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- 108020004635 Complementary DNA Proteins 0.000 description 2
- OHOQEZWSNFNUSY-UHFFFAOYSA-N Cy3-bifunctional dye zwitterion Chemical compound O=C1CCC(=O)N1OC(=O)CCCCCN1C2=CC=C(S(O)(=O)=O)C=C2C(C)(C)C1=CC=CC(C(C1=CC(=CC=C11)S([O-])(=O)=O)(C)C)=[N+]1CCCCCC(=O)ON1C(=O)CCC1=O OHOQEZWSNFNUSY-UHFFFAOYSA-N 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 238000003775 Density Functional Theory Methods 0.000 description 2
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 208000032096 Encephalitic infection Diseases 0.000 description 2
- 241000991587 Enterovirus C Species 0.000 description 2
- 102100023721 Ephrin-B2 Human genes 0.000 description 2
- 108010044090 Ephrin-B2 Proteins 0.000 description 2
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 101000901154 Homo sapiens Complement C3 Proteins 0.000 description 2
- 241000598436 Human T-cell lymphotropic virus Species 0.000 description 2
- 241000701085 Human alphaherpesvirus 3 Species 0.000 description 2
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 208000006142 Infectious Encephalitis Diseases 0.000 description 2
- 102000011781 Karyopherins Human genes 0.000 description 2
- 108010062228 Karyopherins Proteins 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- OFOBLEOULBTSOW-UHFFFAOYSA-L Malonate Chemical compound [O-]C(=O)CC([O-])=O OFOBLEOULBTSOW-UHFFFAOYSA-L 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 241000712079 Measles morbillivirus Species 0.000 description 2
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 2
- MBBZMMPHUWSWHV-BDVNFPICSA-N N-methylglucamine Chemical compound CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO MBBZMMPHUWSWHV-BDVNFPICSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- UFWIBTONFRDIAS-UHFFFAOYSA-N Naphthalene Chemical compound C1=CC=CC2=CC=CC=C21 UFWIBTONFRDIAS-UHFFFAOYSA-N 0.000 description 2
- 241000244206 Nematoda Species 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 108010009711 Phalloidine Proteins 0.000 description 2
- 229920002873 Polyethylenimine Polymers 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 2
- 241000711798 Rabies lyssavirus Species 0.000 description 2
- VYGQUTWHTHXGQB-FFHKNEKCSA-N Retinol Palmitate Chemical compound CCCCCCCCCCCCCCCC(=O)OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C VYGQUTWHTHXGQB-FFHKNEKCSA-N 0.000 description 2
- 241000607768 Shigella Species 0.000 description 2
- 241000700584 Simplexvirus Species 0.000 description 2
- 108091027967 Small hairpin RNA Proteins 0.000 description 2
- 108020004459 Small interfering RNA Proteins 0.000 description 2
- 241000710888 St. Louis encephalitis virus Species 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 229920002253 Tannate Polymers 0.000 description 2
- ZMZDMBWJUHKJPS-UHFFFAOYSA-M Thiocyanate anion Chemical compound [S-]C#N ZMZDMBWJUHKJPS-UHFFFAOYSA-M 0.000 description 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
- 101900005469 Venezuelan equine encephalitis virus Capsid protein Proteins 0.000 description 2
- 241000710772 Yellow fever virus Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- WNLRTRBMVRJNCN-UHFFFAOYSA-L adipate(2-) Chemical compound [O-]C(=O)CCCCC([O-])=O WNLRTRBMVRJNCN-UHFFFAOYSA-L 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 229940072056 alginate Drugs 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 125000004390 alkyl sulfonyl group Chemical group 0.000 description 2
- QYIXCDOBOSTCEI-UHFFFAOYSA-N alpha-cholestanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 QYIXCDOBOSTCEI-UHFFFAOYSA-N 0.000 description 2
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 2
- 229940037003 alum Drugs 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 210000002821 alveolar epithelial cell Anatomy 0.000 description 2
- 239000000908 ammonium hydroxide Substances 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000004596 appetite loss Effects 0.000 description 2
- 125000002102 aryl alkyloxo group Chemical group 0.000 description 2
- 229940072107 ascorbate Drugs 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 229940009098 aspartate Drugs 0.000 description 2
- 229910052788 barium Inorganic materials 0.000 description 2
- DSAJWYNOEDNPEQ-UHFFFAOYSA-N barium atom Chemical compound [Ba] DSAJWYNOEDNPEQ-UHFFFAOYSA-N 0.000 description 2
- 229940077388 benzenesulfonate Drugs 0.000 description 2
- SRSXLGNVWSONIS-UHFFFAOYSA-M benzenesulfonate Chemical compound [O-]S(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-M 0.000 description 2
- 229940050390 benzoate Drugs 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 125000004196 benzothienyl group Chemical group S1C(=CC2=C1C=CC=C2)* 0.000 description 2
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- XMIIGOLPHOKFCH-UHFFFAOYSA-N beta-phenylpropanoic acid Natural products OC(=O)CCC1=CC=CC=C1 XMIIGOLPHOKFCH-UHFFFAOYSA-N 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 229910052797 bismuth Inorganic materials 0.000 description 2
- JCXGWMGPZLAOME-UHFFFAOYSA-N bismuth atom Chemical compound [Bi] JCXGWMGPZLAOME-UHFFFAOYSA-N 0.000 description 2
- 230000008499 blood brain barrier function Effects 0.000 description 2
- 210000001218 blood-brain barrier Anatomy 0.000 description 2
- 235000010354 butylated hydroxytoluene Nutrition 0.000 description 2
- 229940095259 butylated hydroxytoluene Drugs 0.000 description 2
- 238000010804 cDNA synthesis Methods 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- FATUQANACHZLRT-KMRXSBRUSA-L calcium glucoheptonate Chemical compound [Ca+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)C([O-])=O FATUQANACHZLRT-KMRXSBRUSA-L 0.000 description 2
- MIOPJNTWMNEORI-UHFFFAOYSA-N camphorsulfonic acid Chemical compound C1CC2(CS(O)(=O)=O)C(=O)CC1C2(C)C MIOPJNTWMNEORI-UHFFFAOYSA-N 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 239000001768 carboxy methyl cellulose Substances 0.000 description 2
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 2
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- VDANGULDQQJODZ-UHFFFAOYSA-N chloroprocaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1Cl VDANGULDQQJODZ-UHFFFAOYSA-N 0.000 description 2
- 229960002023 chloroprocaine Drugs 0.000 description 2
- ZPEIMTDSQAKGNT-UHFFFAOYSA-N chlorpromazine Chemical compound C1=C(Cl)C=C2N(CCCN(C)C)C3=CC=CC=C3SC2=C1 ZPEIMTDSQAKGNT-UHFFFAOYSA-N 0.000 description 2
- 229960001076 chlorpromazine Drugs 0.000 description 2
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 2
- 229960001231 choline Drugs 0.000 description 2
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@](C)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 description 2
- 230000024203 complement activation Effects 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000009833 condensation Methods 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- 238000004624 confocal microscopy Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 2
- 125000001511 cyclopentyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 2
- 230000000120 cytopathologic effect Effects 0.000 description 2
- 239000000032 diagnostic agent Substances 0.000 description 2
- ACYGYJFTZSAZKR-UHFFFAOYSA-J dicalcium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [Ca+2].[Ca+2].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O ACYGYJFTZSAZKR-UHFFFAOYSA-J 0.000 description 2
- ZBCBWPMODOFKDW-UHFFFAOYSA-N diethanolamine Chemical compound OCCNCCO ZBCBWPMODOFKDW-UHFFFAOYSA-N 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N diphenyl Chemical compound C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 2
- 239000001177 diphosphate Substances 0.000 description 2
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 2
- 235000011180 diphosphates Nutrition 0.000 description 2
- 239000002270 dispersing agent Substances 0.000 description 2
- POULHZVOKOAJMA-UHFFFAOYSA-M dodecanoate Chemical compound CCCCCCCCCCCC([O-])=O POULHZVOKOAJMA-UHFFFAOYSA-M 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 229940009662 edetate Drugs 0.000 description 2
- 238000001493 electron microscopy Methods 0.000 description 2
- 230000009881 electrostatic interaction Effects 0.000 description 2
- 210000001163 endosome Anatomy 0.000 description 2
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 2
- 239000012458 free base Substances 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 229930195712 glutamate Natural products 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 244000000013 helminth Species 0.000 description 2
- 125000003187 heptyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 125000005844 heterocyclyloxy group Chemical group 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-M hexadecanoate Chemical compound CCCCCCCCCCCCCCCC([O-])=O IPCSVZSSVZVIGE-UHFFFAOYSA-M 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- ZMZDMBWJUHKJPS-UHFFFAOYSA-N hydrogen thiocyanate Natural products SC#N ZMZDMBWJUHKJPS-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-M hydrogensulfate Chemical compound OS([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-M 0.000 description 2
- 238000010335 hydrothermal treatment Methods 0.000 description 2
- 238000010191 image analysis Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000002163 immunogen Effects 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 125000001041 indolyl group Chemical group 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 125000001786 isothiazolyl group Chemical group 0.000 description 2
- 229940001447 lactate Drugs 0.000 description 2
- 229940099584 lactobionate Drugs 0.000 description 2
- JYTUSYBCFIZPBE-AMTLMPIISA-N lactobionic acid Chemical compound OC(=O)[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O JYTUSYBCFIZPBE-AMTLMPIISA-N 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 229940070765 laurate Drugs 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 229910052744 lithium Inorganic materials 0.000 description 2
- 235000021266 loss of appetite Nutrition 0.000 description 2
- 208000019017 loss of appetite Diseases 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 229940049920 malate Drugs 0.000 description 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 2
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 2
- IWYDHOAUDWTVEP-UHFFFAOYSA-M mandelate Chemical compound [O-]C(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-M 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 229940102396 methyl bromide Drugs 0.000 description 2
- 229920000609 methyl cellulose Polymers 0.000 description 2
- LRMHVVPPGGOAJQ-UHFFFAOYSA-N methyl nitrate Chemical compound CO[N+]([O-])=O LRMHVVPPGGOAJQ-UHFFFAOYSA-N 0.000 description 2
- JZMJDSHXVKJFKW-UHFFFAOYSA-M methyl sulfate(1-) Chemical compound COS([O-])(=O)=O JZMJDSHXVKJFKW-UHFFFAOYSA-M 0.000 description 2
- 239000001923 methylcellulose Substances 0.000 description 2
- 235000010981 methylcellulose Nutrition 0.000 description 2
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 2
- 210000003632 microfilament Anatomy 0.000 description 2
- 239000007758 minimum essential medium Substances 0.000 description 2
- ZMHHHDYLXOKMOO-UHFFFAOYSA-N n-[4-[[4-[9-[4-[(4-acetamidophenyl)sulfonylamino]phenyl]fluoren-9-yl]phenyl]sulfamoyl]phenyl]acetamide Chemical compound C1=CC(NC(=O)C)=CC=C1S(=O)(=O)NC1=CC=C(C2(C3=CC=CC=C3C3=CC=CC=C32)C=2C=CC(NS(=O)(=O)C=3C=CC(NC(C)=O)=CC=3)=CC=2)C=C1 ZMHHHDYLXOKMOO-UHFFFAOYSA-N 0.000 description 2
- KVBGVZZKJNLNJU-UHFFFAOYSA-M naphthalene-2-sulfonate Chemical compound C1=CC=CC2=CC(S(=O)(=O)[O-])=CC=C21 KVBGVZZKJNLNJU-UHFFFAOYSA-M 0.000 description 2
- 235000001968 nicotinic acid Nutrition 0.000 description 2
- 239000011664 nicotinic acid Substances 0.000 description 2
- 125000004971 nitroalkyl group Chemical group 0.000 description 2
- 231100000956 nontoxicity Toxicity 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 229940049964 oleate Drugs 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 229940123629 p21 activated kinase inhibitor Drugs 0.000 description 2
- JRKICGRDRMAZLK-UHFFFAOYSA-L peroxydisulfate Chemical compound [O-]S(=O)(=O)OOS([O-])(=O)=O JRKICGRDRMAZLK-UHFFFAOYSA-L 0.000 description 2
- 238000002823 phage display Methods 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 125000003170 phenylsulfonyl group Chemical group C1(=CC=CC=C1)S(=O)(=O)* 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 2
- 229940075930 picrate Drugs 0.000 description 2
- OXNIZHLAWKMVMX-UHFFFAOYSA-M picrate anion Chemical compound [O-]C1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-M 0.000 description 2
- 229950010765 pivalate Drugs 0.000 description 2
- IUGYQRQAERSCNH-UHFFFAOYSA-N pivalic acid Chemical compound CC(C)(C)C(O)=O IUGYQRQAERSCNH-UHFFFAOYSA-N 0.000 description 2
- 229920000724 poly(L-arginine) polymer Polymers 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 230000002028 premature Effects 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- MFDFERRIHVXMIY-UHFFFAOYSA-N procaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 MFDFERRIHVXMIY-UHFFFAOYSA-N 0.000 description 2
- 229960004919 procaine Drugs 0.000 description 2
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 2
- 230000005180 public health Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-O pyridinium Chemical compound C1=CC=[NH+]C=C1 JUJWROOIHBZHMG-UHFFFAOYSA-O 0.000 description 2
- 125000000714 pyrimidinyl group Chemical group 0.000 description 2
- 125000000719 pyrrolidinyl group Chemical group 0.000 description 2
- 125000001453 quaternary ammonium group Chemical group 0.000 description 2
- 238000009877 rendering Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-M salicylate Chemical compound OC1=CC=CC=C1C([O-])=O YGSDEFSMJLZEOE-UHFFFAOYSA-M 0.000 description 2
- 229960001860 salicylate Drugs 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 229920002545 silicone oil Polymers 0.000 description 2
- AWUCVROLDVIAJX-GSVOUGTGSA-N sn-glycerol 3-phosphate Chemical compound OC[C@@H](O)COP(O)(O)=O AWUCVROLDVIAJX-GSVOUGTGSA-N 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 229940032147 starch Drugs 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 description 2
- 229940095064 tartrate Drugs 0.000 description 2
- CBXCPBUEXACCNR-UHFFFAOYSA-N tetraethylammonium Chemical compound CC[N+](CC)(CC)CC CBXCPBUEXACCNR-UHFFFAOYSA-N 0.000 description 2
- 125000005958 tetrahydrothienyl group Chemical group 0.000 description 2
- QEMXHQIAXOOASZ-UHFFFAOYSA-N tetramethylammonium Chemical compound C[N+](C)(C)C QEMXHQIAXOOASZ-UHFFFAOYSA-N 0.000 description 2
- 125000001984 thiazolidinyl group Chemical group 0.000 description 2
- 125000005309 thioalkoxy group Chemical group 0.000 description 2
- 150000003573 thiols Chemical class 0.000 description 2
- 230000036962 time dependent Effects 0.000 description 2
- 230000032258 transport Effects 0.000 description 2
- 230000001960 triggered effect Effects 0.000 description 2
- ZDPHROOEEOARMN-UHFFFAOYSA-N undecanoic acid Chemical compound CCCCCCCCCCC(O)=O ZDPHROOEEOARMN-UHFFFAOYSA-N 0.000 description 2
- 241001529453 unidentified herpesvirus Species 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical class CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 2
- QDLHCMPXEPAAMD-QAIWCSMKSA-N wortmannin Chemical compound C1([C@]2(C)C3=C(C4=O)OC=C3C(=O)O[C@@H]2COC)=C4[C@@H]2CCC(=O)[C@@]2(C)C[C@H]1OC(C)=O QDLHCMPXEPAAMD-QAIWCSMKSA-N 0.000 description 2
- 229940051021 yellow-fever virus Drugs 0.000 description 2
- 238000000733 zeta-potential measurement Methods 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- KZJWDPNRJALLNS-VPUBHVLGSA-N (-)-beta-Sitosterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@@H](C(C)C)CC)C)CC4)CC3)CC=2)CC1 KZJWDPNRJALLNS-VPUBHVLGSA-N 0.000 description 1
- BQPPJGMMIYJVBR-UHFFFAOYSA-N (10S)-3c-Acetoxy-4.4.10r.13c.14t-pentamethyl-17c-((R)-1.5-dimethyl-hexen-(4)-yl)-(5tH)-Delta8-tetradecahydro-1H-cyclopenta[a]phenanthren Natural products CC12CCC(OC(C)=O)C(C)(C)C1CCC1=C2CCC2(C)C(C(CCC=C(C)C)C)CCC21C BQPPJGMMIYJVBR-UHFFFAOYSA-N 0.000 description 1
- CSVWWLUMXNHWSU-UHFFFAOYSA-N (22E)-(24xi)-24-ethyl-5alpha-cholest-22-en-3beta-ol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(CC)C(C)C)C1(C)CC2 CSVWWLUMXNHWSU-UHFFFAOYSA-N 0.000 description 1
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 description 1
- VGSSUFQMXBFFTM-UHFFFAOYSA-N (24R)-24-ethyl-5alpha-cholestane-3beta,5,6beta-triol Natural products C1C(O)C2(O)CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 VGSSUFQMXBFFTM-UHFFFAOYSA-N 0.000 description 1
- LNAZSHAWQACDHT-XIYTZBAFSA-N (2r,3r,4s,5r,6s)-4,5-dimethoxy-2-(methoxymethyl)-3-[(2s,3r,4s,5r,6r)-3,4,5-trimethoxy-6-(methoxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6r)-4,5,6-trimethoxy-2-(methoxymethyl)oxan-3-yl]oxyoxane Chemical compound CO[C@@H]1[C@@H](OC)[C@H](OC)[C@@H](COC)O[C@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@H](O[C@H]2[C@@H]([C@@H](OC)[C@H](OC)O[C@@H]2COC)OC)O[C@@H]1COC LNAZSHAWQACDHT-XIYTZBAFSA-N 0.000 description 1
- XUNKPNYCNUKOAU-VXJRNSOOSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-amino-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]a Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O XUNKPNYCNUKOAU-VXJRNSOOSA-N 0.000 description 1
- RAVVEEJGALCVIN-AGVBWZICSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-5-amino-2-[[(2s)-2-[[(2s)-2-[[(2s)-6-amino-2-[[(2s)-6-amino-2-[[(2s)-2-[[2-[[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]acetyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]hexanoyl]amino]hexanoyl]amino]-5-(diamino Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 RAVVEEJGALCVIN-AGVBWZICSA-N 0.000 description 1
- IZVFFXVYBHFIHY-UHFFFAOYSA-N (3alpha, 5alpha)-Cholest-7-en-3-ol, 9CI Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)CCCC(C)C)CCC33)C)C3=CCC21 IZVFFXVYBHFIHY-UHFFFAOYSA-N 0.000 description 1
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 1
- CHGIKSSZNBCNDW-UHFFFAOYSA-N (3beta,5alpha)-4,4-Dimethylcholesta-8,24-dien-3-ol Natural products CC12CCC(O)C(C)(C)C1CCC1=C2CCC2(C)C(C(CCC=C(C)C)C)CCC21 CHGIKSSZNBCNDW-UHFFFAOYSA-N 0.000 description 1
- VOZOGDCWCKQPMK-NFFVHWSESA-N (6z)-3-[2-[4-(2-methoxyphenyl)piperazin-1-yl]ethoxy]-6-(4-phenyl-1,2-dihydropyrazol-3-ylidene)cyclohexa-2,4-dien-1-one Chemical compound COC1=CC=CC=C1N1CCN(CCOC=2C=CC(/C(=O)C=2)=C/2C(=CNN\2)C=2C=CC=CC=2)CC1 VOZOGDCWCKQPMK-NFFVHWSESA-N 0.000 description 1
- 125000000171 (C1-C6) haloalkyl group Chemical group 0.000 description 1
- CITHEXJVPOWHKC-UUWRZZSWSA-N 1,2-di-O-myristoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCC CITHEXJVPOWHKC-UUWRZZSWSA-N 0.000 description 1
- SLKDGVPOSSLUAI-PGUFJCEWSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine zwitterion Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OCCN)OC(=O)CCCCCCCCCCCCCCC SLKDGVPOSSLUAI-PGUFJCEWSA-N 0.000 description 1
- IJFVSSZAOYLHEE-SSEXGKCCSA-N 1,2-dilauroyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCC IJFVSSZAOYLHEE-SSEXGKCCSA-N 0.000 description 1
- UHUSDOQQWJGJQS-QNGWXLTQSA-N 1,2-dioctadecanoyl-sn-glycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](CO)OC(=O)CCCCCCCCCCCCCCCCC UHUSDOQQWJGJQS-QNGWXLTQSA-N 0.000 description 1
- DSNRWDQKZIEDDB-SQYFZQSCSA-N 1,2-dioleoyl-sn-glycero-3-phospho-(1'-sn-glycerol) Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@@H](O)CO)OC(=O)CCCCCCC\C=C/CCCCCCCC DSNRWDQKZIEDDB-SQYFZQSCSA-N 0.000 description 1
- WTBFLCSPLLEDEM-JIDRGYQWSA-N 1,2-dioleoyl-sn-glycero-3-phospho-L-serine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCC\C=C/CCCCCCCC WTBFLCSPLLEDEM-JIDRGYQWSA-N 0.000 description 1
- MWRBNPKJOOWZPW-NYVOMTAGSA-N 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine zwitterion Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP(O)(=O)OCCN)OC(=O)CCCCCCC\C=C/CCCCCCCC MWRBNPKJOOWZPW-NYVOMTAGSA-N 0.000 description 1
- AFSHUZFNMVJNKX-LLWMBOQKSA-N 1,2-dioleoyl-sn-glycerol Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](CO)OC(=O)CCCCCCC\C=C/CCCCCCCC AFSHUZFNMVJNKX-LLWMBOQKSA-N 0.000 description 1
- JEJLGIQLPYYGEE-XIFFEERXSA-N 1,2-dipalmitoyl-sn-glycerol Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](CO)OC(=O)CCCCCCCCCCCCCCC JEJLGIQLPYYGEE-XIFFEERXSA-N 0.000 description 1
- JFBCSFJKETUREV-LJAQVGFWSA-N 1,2-ditetradecanoyl-sn-glycerol Chemical compound CCCCCCCCCCCCCC(=O)OC[C@H](CO)OC(=O)CCCCCCCCCCCCC JFBCSFJKETUREV-LJAQVGFWSA-N 0.000 description 1
- WTJKGGKOPKCXLL-VYOBOKEXSA-N 1-hexadecanoyl-2-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC WTJKGGKOPKCXLL-VYOBOKEXSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 150000000252 12-deoxycholic acids Chemical class 0.000 description 1
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
- XYTLYKGXLMKYMV-UHFFFAOYSA-N 14alpha-methylzymosterol Natural products CC12CCC(O)CC1CCC1=C2CCC2(C)C(C(CCC=C(C)C)C)CCC21C XYTLYKGXLMKYMV-UHFFFAOYSA-N 0.000 description 1
- LDGWQMRUWMSZIU-LQDDAWAPSA-M 2,3-bis[(z)-octadec-9-enoxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCC\C=C/CCCCCCCC LDGWQMRUWMSZIU-LQDDAWAPSA-M 0.000 description 1
- NTTZBBIBMSBLNK-UHFFFAOYSA-M 2,3-di(octadecanoyloxy)propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCCCC NTTZBBIBMSBLNK-UHFFFAOYSA-M 0.000 description 1
- BFTWAJQCIAJTEA-UHFFFAOYSA-N 2-(1,3-diazinan-1-ylmethyl)-6-nitro-3-phenylquinazolin-4-one Chemical compound C=1C=CC=CC=1N1C(=O)C2=CC([N+](=O)[O-])=CC=C2N=C1CN1CCCNC1 BFTWAJQCIAJTEA-UHFFFAOYSA-N 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- ZDCQUCHJJUYSMO-UHFFFAOYSA-N 2-(3-fluorothiophen-2-yl)-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2-yl)acetamide Chemical compound S1C(=NC2=C1CCCC2)NC(CC=1SC=CC=1F)=O ZDCQUCHJJUYSMO-UHFFFAOYSA-N 0.000 description 1
- RNAMYOYQYRYFQY-UHFFFAOYSA-N 2-(4,4-difluoropiperidin-1-yl)-6-methoxy-n-(1-propan-2-ylpiperidin-4-yl)-7-(3-pyrrolidin-1-ylpropoxy)quinazolin-4-amine Chemical compound N1=C(N2CCC(F)(F)CC2)N=C2C=C(OCCCN3CCCC3)C(OC)=CC2=C1NC1CCN(C(C)C)CC1 RNAMYOYQYRYFQY-UHFFFAOYSA-N 0.000 description 1
- ZKBCZWVZUYRBJL-UHFFFAOYSA-N 2-(4-phenylmethoxyanilino)pyrido[3,2-e][1,3]thiazin-4-one Chemical compound S1C2=NC=CC=C2C(=O)N=C1NC(C=C1)=CC=C1OCC1=CC=CC=C1 ZKBCZWVZUYRBJL-UHFFFAOYSA-N 0.000 description 1
- HMRRSMNUURYCSB-UHFFFAOYSA-N 2-(n-methyl-4-phenylmethoxyanilino)pyrido[3,2-e][1,3]thiazin-4-one Chemical compound N=1C(=O)C2=CC=CN=C2SC=1N(C)C(C=C1)=CC=C1OCC1=CC=CC=C1 HMRRSMNUURYCSB-UHFFFAOYSA-N 0.000 description 1
- NPEFAIPQXBJTAS-UHFFFAOYSA-N 2-[(1-ethyl-4-methylpiperazin-2-ylidene)amino]-5-nitro-n-phenylbenzamide Chemical compound CCN1CCN(C)CC1=NC1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=CC=C1 NPEFAIPQXBJTAS-UHFFFAOYSA-N 0.000 description 1
- PNJBOAVCVAVRGR-UDCAXGDQSA-N 22-Hopanol Chemical compound C([C@]1(C)[C@H]2CC[C@H]34)CCC(C)(C)[C@@H]1CC[C@@]2(C)[C@]4(C)CC[C@@H]1[C@]3(C)CC[C@@H]1C(C)(O)C PNJBOAVCVAVRGR-UDCAXGDQSA-N 0.000 description 1
- ARYTXMNEANMLMU-UHFFFAOYSA-N 24alpha-methylcholestanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(C)C(C)C)C1(C)CC2 ARYTXMNEANMLMU-UHFFFAOYSA-N 0.000 description 1
- KLEXDBGYSOIREE-UHFFFAOYSA-N 24xi-n-propylcholesterol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CCC)C(C)C)C1(C)CC2 KLEXDBGYSOIREE-UHFFFAOYSA-N 0.000 description 1
- NWDBLYBNYAZJBL-UHFFFAOYSA-N 3-(3,5-dimethyl-1,2-oxazol-4-yl)-N-[4-(2-oxo-1,3-dihydroindol-5-yl)-1,3-thiazol-2-yl]prop-2-enamide Chemical compound CC1=NOC(=C1C=CC(=O)NC=1SC=C(N=1)C=1C=C2CC(NC2=CC=1)=O)C NWDBLYBNYAZJBL-UHFFFAOYSA-N 0.000 description 1
- FPTJELQXIUUCEY-UHFFFAOYSA-N 3beta-Hydroxy-lanostan Natural products C1CC2C(C)(C)C(O)CCC2(C)C2C1C1(C)CCC(C(C)CCCC(C)C)C1(C)CC2 FPTJELQXIUUCEY-UHFFFAOYSA-N 0.000 description 1
- LFQXEZVYNCBVDO-PBJLWWPKSA-N 4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol Chemical compound C([C@@]12C)C[C@H](O)C(C)(C)[C@@H]1CCC1=C2CC[C@]2(C)[C@@H]([C@@H](CCC=C(C)C)C)CC=C21 LFQXEZVYNCBVDO-PBJLWWPKSA-N 0.000 description 1
- UAZJPMMKWBPACD-GCXDCGAKSA-N 4-amino-1-[(1r,2s,3r,4r)-2,3-dihydroxy-4-(hydroxymethyl)cyclopentyl]pyrimidin-2-one Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)C1 UAZJPMMKWBPACD-GCXDCGAKSA-N 0.000 description 1
- IZVFFXVYBHFIHY-SKCNUYALSA-N 5alpha-cholest-7-en-3beta-ol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@@H](CC[C@@]3([C@@H]([C@H](C)CCCC(C)C)CC[C@H]33)C)C3=CC[C@H]21 IZVFFXVYBHFIHY-SKCNUYALSA-N 0.000 description 1
- QETLKNDKQOXZRP-XTGBIJOFSA-N 5alpha-cholest-8-en-3beta-ol Chemical compound C([C@@]12C)C[C@H](O)C[C@@H]1CCC1=C2CC[C@]2(C)[C@@H]([C@H](C)CCCC(C)C)CC[C@H]21 QETLKNDKQOXZRP-XTGBIJOFSA-N 0.000 description 1
- QETLKNDKQOXZRP-UHFFFAOYSA-N 5alpha-cholest-8-en-3beta-ol Natural products CC12CCC(O)CC1CCC1=C2CCC2(C)C(C(C)CCCC(C)C)CCC21 QETLKNDKQOXZRP-UHFFFAOYSA-N 0.000 description 1
- VHAMERSKVUBMLK-UHFFFAOYSA-N 6,7-difluoro-3-phenyl-2-(piperazin-1-ylmethyl)quinazolin-4-one Chemical compound C=1C=CC=CC=1N1C(=O)C=2C=C(F)C(F)=CC=2N=C1CN1CCNCC1 VHAMERSKVUBMLK-UHFFFAOYSA-N 0.000 description 1
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 description 1
- RUSSPKPUXDSHNC-DDPQNLDTSA-N 7-dehydrodesmosterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@@H](CC[C@@]3([C@@H]([C@@H](CCC=C(C)C)C)CC[C@H]33)C)C3=CC=C21 RUSSPKPUXDSHNC-DDPQNLDTSA-N 0.000 description 1
- 229940121819 ATPase inhibitor Drugs 0.000 description 1
- 241000224422 Acanthamoeba Species 0.000 description 1
- 241000701242 Adenoviridae Species 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- 239000012114 Alexa Fluor 647 Substances 0.000 description 1
- 206010001935 American trypanosomiasis Diseases 0.000 description 1
- 241000498253 Ancylostoma duodenale Species 0.000 description 1
- 108700042778 Antimicrobial Peptides Proteins 0.000 description 1
- 102000044503 Antimicrobial Peptides Human genes 0.000 description 1
- 101710081722 Antitrypsin Proteins 0.000 description 1
- 241000712892 Arenaviridae Species 0.000 description 1
- 241000712891 Arenavirus Species 0.000 description 1
- 241000244185 Ascaris lumbricoides Species 0.000 description 1
- 206010003591 Ataxia Diseases 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000193738 Bacillus anthracis Species 0.000 description 1
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000589562 Brucella Species 0.000 description 1
- 241000589567 Brucella abortus Species 0.000 description 1
- 241001148106 Brucella melitensis Species 0.000 description 1
- 241001148111 Brucella suis Species 0.000 description 1
- 241001453380 Burkholderia Species 0.000 description 1
- 241000722910 Burkholderia mallei Species 0.000 description 1
- 125000006577 C1-C6 hydroxyalkyl group Chemical group 0.000 description 1
- JRGGPBYAPMLJDO-RELWKKBWSA-N C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=C(F)C=C1 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=C(F)C=C1 JRGGPBYAPMLJDO-RELWKKBWSA-N 0.000 description 1
- IDFAXXBVRXTORW-RELWKKBWSA-N C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=CC(F)=C1 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=CC(F)=C1 IDFAXXBVRXTORW-RELWKKBWSA-N 0.000 description 1
- DPNJQHLBYVBVRJ-KNTRCKAVSA-N C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CSC=C1 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CSC=C1 DPNJQHLBYVBVRJ-KNTRCKAVSA-N 0.000 description 1
- LAHSLFBEBUFIMG-LYBHJNIJSA-N C1N(C)CCN(C)\C1=N\c1cc(F)c(C#N)cc1C(=O)Nc1ccccc1 Chemical compound C1N(C)CCN(C)\C1=N\c1cc(F)c(C#N)cc1C(=O)Nc1ccccc1 LAHSLFBEBUFIMG-LYBHJNIJSA-N 0.000 description 1
- HHWQACWELJHXAV-PTGBLXJZSA-N C1N(C)CCN(C)\C1=N\c1cc(F)c(F)cc1C(=O)Nc1ccccc1 Chemical compound C1N(C)CCN(C)\C1=N\c1cc(F)c(F)cc1C(=O)Nc1ccccc1 HHWQACWELJHXAV-PTGBLXJZSA-N 0.000 description 1
- XNDWUHNSNYTVSQ-FCDQGJHFSA-N C1N(C)CCN(C)\C1=N\c1ccc(C#N)cc1C(=O)Nc1ccccc1F Chemical compound C1N(C)CCN(C)\C1=N\c1ccc(C#N)cc1C(=O)Nc1ccccc1F XNDWUHNSNYTVSQ-FCDQGJHFSA-N 0.000 description 1
- PTSWNHNLFOVFMF-XIEYBQDHSA-N C1N(C)CCN(C)\C1=N\c1ccc(C(F)(F)F)cc1C(=O)Nc1ccccc1 Chemical compound C1N(C)CCN(C)\C1=N\c1ccc(C(F)(F)F)cc1C(=O)Nc1ccccc1 PTSWNHNLFOVFMF-XIEYBQDHSA-N 0.000 description 1
- CUJIUPBIPVFSII-ZBJSNUHESA-N C1N(C)CCN(C)\C1=N\c1ccc([N+]([O-])=O)cc1C(=O)NCc1ccccc1 Chemical compound C1N(C)CCN(C)\C1=N\c1ccc([N+]([O-])=O)cc1C(=O)NCc1ccccc1 CUJIUPBIPVFSII-ZBJSNUHESA-N 0.000 description 1
- YZHGVGQLCNDECX-HMMYKYKNSA-N C1N(C)CCN(C)\C1=N\c1ccnnc1C(=O)Nc1ccccc1 Chemical compound C1N(C)CCN(C)\C1=N\c1ccnnc1C(=O)Nc1ccccc1 YZHGVGQLCNDECX-HMMYKYKNSA-N 0.000 description 1
- 125000000041 C6-C10 aryl group Chemical group 0.000 description 1
- UTPWYBBWALNFIF-FCDQGJHFSA-N C=1C=CC=CC=1NC(=O)C1=CC(C(=O)OC)=CC=C1\N=C1/CN(C)CCN1C Chemical compound C=1C=CC=CC=1NC(=O)C1=CC(C(=O)OC)=CC=C1\N=C1/CN(C)CCN1C UTPWYBBWALNFIF-FCDQGJHFSA-N 0.000 description 1
- UUCCXUKLRYEGBO-OBGWFSINSA-N CC(C)NC(=O)c1cc([N+]([O-])=O)ccc1\N=C1/CN(C)CCN1C Chemical compound CC(C)NC(=O)c1cc([N+]([O-])=O)ccc1\N=C1/CN(C)CCN1C UUCCXUKLRYEGBO-OBGWFSINSA-N 0.000 description 1
- 102100022002 CD59 glycoprotein Human genes 0.000 description 1
- WPCRCMLSISGKQV-UHFFFAOYSA-N CN1C(CN(CC1)C)=NC1=C(C(=O)NC2=C(C=CC=C2)OC)C=C(C=C1)[N+](=O)[O-] Chemical compound CN1C(CN(CC1)C)=NC1=C(C(=O)NC2=C(C=CC=C2)OC)C=C(C=C1)[N+](=O)[O-] WPCRCMLSISGKQV-UHFFFAOYSA-N 0.000 description 1
- WPWNJTFIHUGULG-DTQAZKPQSA-N CNC(=O)c1cc([N+]([O-])=O)ccc1\N=C1/CN(C)CCN1C Chemical compound CNC(=O)c1cc([N+]([O-])=O)ccc1\N=C1/CN(C)CCN1C WPWNJTFIHUGULG-DTQAZKPQSA-N 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- 241001493160 California encephalitis virus Species 0.000 description 1
- SGNBVLSWZMBQTH-FGAXOLDCSA-N Campesterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@H](C(C)C)C)C)CC4)CC3)CC=2)CC1 SGNBVLSWZMBQTH-FGAXOLDCSA-N 0.000 description 1
- 241000393548 Candidae Species 0.000 description 1
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 241000242722 Cestoda Species 0.000 description 1
- 201000009182 Chikungunya Diseases 0.000 description 1
- 241001647378 Chlamydia psittaci Species 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- BHYOQNUELFTYRT-UHFFFAOYSA-N Cholesterol sulfate Natural products C1C=C2CC(OS(O)(=O)=O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 BHYOQNUELFTYRT-UHFFFAOYSA-N 0.000 description 1
- 239000004380 Cholic acid Substances 0.000 description 1
- LPZCCMIISIBREI-MTFRKTCUSA-N Citrostadienol Natural products CC=C(CC[C@@H](C)[C@H]1CC[C@H]2C3=CC[C@H]4[C@H](C)[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C)C(C)C LPZCCMIISIBREI-MTFRKTCUSA-N 0.000 description 1
- 241000193155 Clostridium botulinum Species 0.000 description 1
- 241000193468 Clostridium perfringens Species 0.000 description 1
- 241000223205 Coccidioides immitis Species 0.000 description 1
- 108700040183 Complement C1 Inhibitor Proteins 0.000 description 1
- 102000055157 Complement C1 Inhibitor Human genes 0.000 description 1
- 102100025680 Complement decay-accelerating factor Human genes 0.000 description 1
- 102100030886 Complement receptor type 1 Human genes 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 241000711573 Coronaviridae Species 0.000 description 1
- 241000186216 Corynebacterium Species 0.000 description 1
- 241000606678 Coxiella burnetii Species 0.000 description 1
- 229920002785 Croscarmellose sodium Polymers 0.000 description 1
- 239000004971 Cross linker Substances 0.000 description 1
- 241000223936 Cryptosporidium parvum Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 241000450599 DNA viruses Species 0.000 description 1
- 206010011906 Death Diseases 0.000 description 1
- ARVGMISWLZPBCH-UHFFFAOYSA-N Dehydro-beta-sitosterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)CCC(CC)C(C)C)CCC33)C)C3=CC=C21 ARVGMISWLZPBCH-UHFFFAOYSA-N 0.000 description 1
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- 229940122809 Dynamin 2 inhibitor Drugs 0.000 description 1
- 102000020630 Dynamin II Human genes 0.000 description 1
- 108010044191 Dynamin II Proteins 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 241001115402 Ebolavirus Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000224432 Entamoeba histolytica Species 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000709661 Enterovirus Species 0.000 description 1
- 102000030797 EphB4 Receptor Human genes 0.000 description 1
- 108010055323 EphB4 Receptor Proteins 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000495778 Escherichia faecalis Species 0.000 description 1
- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical compound CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 241000711950 Filoviridae Species 0.000 description 1
- 241000710781 Flaviviridae Species 0.000 description 1
- 241000710831 Flavivirus Species 0.000 description 1
- 241000589602 Francisella tularensis Species 0.000 description 1
- 229910052688 Gadolinium Inorganic materials 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000224467 Giardia intestinalis Species 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- BKLIAINBCQPSOV-UHFFFAOYSA-N Gluanol Natural products CC(C)CC=CC(C)C1CCC2(C)C3=C(CCC12C)C4(C)CCC(O)C(C)(C)C4CC3 BKLIAINBCQPSOV-UHFFFAOYSA-N 0.000 description 1
- 102000019058 Glycogen Synthase Kinase 3 beta Human genes 0.000 description 1
- 108010051975 Glycogen Synthase Kinase 3 beta Proteins 0.000 description 1
- 206010018691 Granuloma Diseases 0.000 description 1
- 108020005004 Guide RNA Proteins 0.000 description 1
- BTEISVKTSQLKST-UHFFFAOYSA-N Haliclonasterol Natural products CC(C=CC(C)C(C)(C)C)C1CCC2C3=CC=C4CC(O)CCC4(C)C3CCC12C BTEISVKTSQLKST-UHFFFAOYSA-N 0.000 description 1
- 108010034145 Helminth Proteins Proteins 0.000 description 1
- 208000019645 Hemorrhagic fever-renal syndrome Diseases 0.000 description 1
- 241000035314 Henipavirus Species 0.000 description 1
- 241000711549 Hepacivirus C Species 0.000 description 1
- 241000700739 Hepadnaviridae Species 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 241000709721 Hepatovirus A Species 0.000 description 1
- 241000700586 Herpesviridae Species 0.000 description 1
- 101000897400 Homo sapiens CD59 glycoprotein Proteins 0.000 description 1
- 101000856022 Homo sapiens Complement decay-accelerating factor Proteins 0.000 description 1
- 101000868279 Homo sapiens Leukocyte surface antigen CD47 Proteins 0.000 description 1
- 101000961414 Homo sapiens Membrane cofactor protein Proteins 0.000 description 1
- 101000987581 Homo sapiens Perforin-1 Proteins 0.000 description 1
- 108700000788 Human immunodeficiency virus 1 tat peptide (47-57) Proteins 0.000 description 1
- 108700003968 Human immunodeficiency virus 1 tat peptide (49-57) Proteins 0.000 description 1
- DLTTWXWDLCGBRD-UHFFFAOYSA-N Hydroxyhopane Natural products CC12CCCC(C)(C)C1CCC1(C)C2CCC2C3(C)CCC(C(C)C)C3(O)CCC21C DLTTWXWDLCGBRD-UHFFFAOYSA-N 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- 206010021079 Hypopnoea Diseases 0.000 description 1
- 102000009490 IgG Receptors Human genes 0.000 description 1
- 108010073807 IgG Receptors Proteins 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 206010022004 Influenza like illness Diseases 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- LOPKHWOTGJIQLC-UHFFFAOYSA-N Lanosterol Natural products CC(CCC=C(C)C)C1CCC2(C)C3=C(CCC12C)C4(C)CCC(C)(O)C(C)(C)C4CC3 LOPKHWOTGJIQLC-UHFFFAOYSA-N 0.000 description 1
- 241000222722 Leishmania <genus> Species 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 102100032913 Leukocyte surface antigen CD47 Human genes 0.000 description 1
- 241000186781 Listeria Species 0.000 description 1
- 241000186779 Listeria monocytogenes Species 0.000 description 1
- 241000712899 Lymphocytic choriomeningitis mammarenavirus Species 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241000712898 Machupo mammarenavirus Species 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001115401 Marburgvirus Species 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102100039373 Membrane cofactor protein Human genes 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 241000711386 Mumps virus Species 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 241000202936 Mycoplasma mycoides Species 0.000 description 1
- 231100000678 Mycotoxin Toxicity 0.000 description 1
- 108091061960 Naked DNA Proteins 0.000 description 1
- 241000737052 Naso hexacanthus Species 0.000 description 1
- 241000498270 Necator americanus Species 0.000 description 1
- CAHGCLMLTWQZNJ-UHFFFAOYSA-N Nerifoliol Natural products CC12CCC(O)C(C)(C)C1CCC1=C2CCC2(C)C(C(CCC=C(C)C)C)CCC21C CAHGCLMLTWQZNJ-UHFFFAOYSA-N 0.000 description 1
- 208000009869 Neu-Laxova syndrome Diseases 0.000 description 1
- 102000001839 Neurturin Human genes 0.000 description 1
- 108010015406 Neurturin Proteins 0.000 description 1
- 101710144128 Non-structural protein 2 Proteins 0.000 description 1
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 1
- 101710199667 Nuclear export protein Proteins 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 102000002488 Nucleoplasmin Human genes 0.000 description 1
- 241000713112 Orthobunyavirus Species 0.000 description 1
- 241000150452 Orthohantavirus Species 0.000 description 1
- 241000712464 Orthomyxoviridae Species 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 241000711504 Paramyxoviridae Species 0.000 description 1
- 208000002606 Paramyxoviridae Infections Diseases 0.000 description 1
- 241000701945 Parvoviridae Species 0.000 description 1
- 229920001774 Perfluoroether Polymers 0.000 description 1
- 241000150350 Peribunyaviridae Species 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000709664 Picornaviridae Species 0.000 description 1
- 102100027637 Plasma protease C1 inhibitor Human genes 0.000 description 1
- 241000224016 Plasmodium Species 0.000 description 1
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 1
- 229920002535 Polyethylene Glycol 1500 Polymers 0.000 description 1
- 229920000604 Polyethylene Glycol 200 Polymers 0.000 description 1
- 229920002582 Polyethylene Glycol 600 Polymers 0.000 description 1
- 241001631648 Polyomaviridae Species 0.000 description 1
- 241000710884 Powassan virus Species 0.000 description 1
- 241000700625 Poxviridae Species 0.000 description 1
- ORNBQBCIOKFOEO-YQUGOWONSA-N Pregnenolone Natural products O=C(C)[C@@H]1[C@@]2(C)[C@H]([C@H]3[C@@H]([C@]4(C)C(=CC3)C[C@@H](O)CC4)CC2)CC1 ORNBQBCIOKFOEO-YQUGOWONSA-N 0.000 description 1
- 101710149951 Protein Tat Proteins 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 238000004617 QSAR study Methods 0.000 description 1
- 108010086344 RALA peptide Proteins 0.000 description 1
- 230000004570 RNA-binding Effects 0.000 description 1
- 241000702247 Reoviridae Species 0.000 description 1
- 241000702263 Reovirus sp. Species 0.000 description 1
- 241000725643 Respiratory syncytial virus Species 0.000 description 1
- 241000712907 Retroviridae Species 0.000 description 1
- 241000711931 Rhabdoviridae Species 0.000 description 1
- 102000004389 Ribonucleoproteins Human genes 0.000 description 1
- 108010081734 Ribonucleoproteins Proteins 0.000 description 1
- 241000606701 Rickettsia Species 0.000 description 1
- 241000606697 Rickettsia prowazekii Species 0.000 description 1
- 241000606695 Rickettsia rickettsii Species 0.000 description 1
- 241000606726 Rickettsia typhi Species 0.000 description 1
- 241000702670 Rotavirus Species 0.000 description 1
- 241000710799 Rubella virus Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 101000910357 Scorpio palmatus Maurocalcin Proteins 0.000 description 1
- 229920001800 Shellac Polymers 0.000 description 1
- 241000607764 Shigella dysenteriae Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- LGJMUZUPVCAVPU-JFBKYFIKSA-N Sitostanol Natural products O[C@@H]1C[C@H]2[C@@](C)([C@@H]3[C@@H]([C@H]4[C@@](C)([C@@H]([C@@H](CC[C@H](C(C)C)CC)C)CC4)CC3)CC2)CC1 LGJMUZUPVCAVPU-JFBKYFIKSA-N 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- QTENRWWVYAAPBI-YZTFXSNBSA-N Streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@H]1[C@H](N=C(N)N)[C@@H](O)[C@H](N=C(N)N)[C@@H](O)[C@@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@H]1[C@H](N=C(N)N)[C@@H](O)[C@H](N=C(N)N)[C@@H](O)[C@@H]1O QTENRWWVYAAPBI-YZTFXSNBSA-N 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 241000906446 Theraps Species 0.000 description 1
- 241000710924 Togaviridae Species 0.000 description 1
- 206010070863 Toxicity to various agents Diseases 0.000 description 1
- 241000223997 Toxoplasma gondii Species 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000242541 Trematoda Species 0.000 description 1
- 241000869417 Trematodes Species 0.000 description 1
- 241000224527 Trichomonas vaginalis Species 0.000 description 1
- 241001489145 Trichuris trichiura Species 0.000 description 1
- DWCSNWXARWMZTG-UHFFFAOYSA-N Trigonegenin A Natural products CC1C(C2(CCC3C4(C)CCC(O)C=C4CCC3C2C2)C)C2OC11CCC(C)CO1 DWCSNWXARWMZTG-UHFFFAOYSA-N 0.000 description 1
- 241000223104 Trypanosoma Species 0.000 description 1
- 241000223105 Trypanosoma brucei Species 0.000 description 1
- 241000223109 Trypanosoma cruzi Species 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 238000010162 Tukey test Methods 0.000 description 1
- HZYXFRGVBOPPNZ-UHFFFAOYSA-N UNPD88870 Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)=CCC(CC)C(C)C)C1(C)CC2 HZYXFRGVBOPPNZ-UHFFFAOYSA-N 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- 241000700647 Variola virus Species 0.000 description 1
- 208000002687 Venezuelan Equine Encephalomyelitis Diseases 0.000 description 1
- 201000009145 Venezuelan equine encephalitis Diseases 0.000 description 1
- 241000607598 Vibrio Species 0.000 description 1
- 241000607626 Vibrio cholerae Species 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 201000006449 West Nile encephalitis Diseases 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 241000607734 Yersinia <bacteria> Species 0.000 description 1
- 241000607447 Yersinia enterocolitica Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- UJELMAYUQSGICC-UHFFFAOYSA-N Zymosterol Natural products CC12CCC(O)CC1CCC1=C2CCC2(C)C(C(C)C=CCC(C)C)CCC21 UJELMAYUQSGICC-UHFFFAOYSA-N 0.000 description 1
- ORIATWTXJGGODP-PSXMRANNSA-N [(2r)-3-hexadecanoyloxy-2-[11-[methyl-(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]undecanoyloxy]propyl] 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCN(C)C1=CC=C([N+]([O-])=O)C2=NON=C12 ORIATWTXJGGODP-PSXMRANNSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- VJHCJDRQFCCTHL-UHFFFAOYSA-N acetic acid 2,3,4,5,6-pentahydroxyhexanal Chemical compound CC(O)=O.OCC(O)C(O)C(O)C(O)C=O VJHCJDRQFCCTHL-UHFFFAOYSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 125000002015 acyclic group Chemical group 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000000362 adenosine triphosphatase inhibitor Substances 0.000 description 1
- 108700010877 adenoviridae proteins Proteins 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000012387 aerosolization Methods 0.000 description 1
- 101150084233 ago2 gene Proteins 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- 125000004453 alkoxycarbonyl group Chemical group 0.000 description 1
- 125000004644 alkyl sulfinyl group Chemical group 0.000 description 1
- 150000008052 alkyl sulfonates Chemical class 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 102000009899 alpha Karyopherins Human genes 0.000 description 1
- 108010077099 alpha Karyopherins Proteins 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 210000001132 alveolar macrophage Anatomy 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 125000006620 amino-(C1-C6) alkyl group Chemical group 0.000 description 1
- 125000004103 aminoalkyl group Chemical group 0.000 description 1
- CAMXVZOXBADHNJ-UHFFFAOYSA-N ammonium nitrite Chemical compound [NH4+].[O-]N=O CAMXVZOXBADHNJ-UHFFFAOYSA-N 0.000 description 1
- 230000000181 anti-adherent effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000003911 antiadherent Substances 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 125000001204 arachidyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 125000005161 aryl oxy carbonyl group Chemical group 0.000 description 1
- 125000005228 aryl sulfonate group Chemical group 0.000 description 1
- 125000004391 aryl sulfonyl group Chemical group 0.000 description 1
- 125000004104 aryloxy group Chemical group 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 125000003725 azepanyl group Chemical group 0.000 description 1
- 125000002393 azetidinyl group Chemical group 0.000 description 1
- 125000005335 azido alkyl group Chemical group 0.000 description 1
- 125000004069 aziridinyl group Chemical group 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 210000004666 bacterial spore Anatomy 0.000 description 1
- 229930192649 bafilomycin Natural products 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 125000003785 benzimidazolyl group Chemical group N1=C(NC2=C1C=CC=C2)* 0.000 description 1
- 125000000499 benzofuranyl group Chemical group O1C(=CC2=C1C=CC=C2)* 0.000 description 1
- 125000004618 benzofuryl group Chemical group O1C(=CC2=C1C=CC=C2)* 0.000 description 1
- 125000001164 benzothiazolyl group Chemical group S1C(=NC2=C1C=CC=C2)* 0.000 description 1
- 125000004541 benzoxazolyl group Chemical group O1C(=NC2=C1C=CC=C2)* 0.000 description 1
- MJVXAPPOFPTTCA-UHFFFAOYSA-N beta-Sistosterol Natural products CCC(CCC(C)C1CCC2C3CC=C4C(C)C(O)CCC4(C)C3CCC12C)C(C)C MJVXAPPOFPTTCA-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- NJKOMDUNNDKEAI-UHFFFAOYSA-N beta-sitosterol Natural products CCC(CCC(C)C1CCC2(C)C3CC=C4CC(O)CCC4C3CCC12C)C(C)C NJKOMDUNNDKEAI-UHFFFAOYSA-N 0.000 description 1
- 125000002619 bicyclic group Chemical group 0.000 description 1
- 231100000693 bioaccumulation Toxicity 0.000 description 1
- 239000012867 bioactive agent Substances 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- CODDAXSDXHVTSW-ZBJSNUHESA-N c1c(OC)cccc1NC(=O)c1cc([N+]([O-])=O)ccc1\N=C1/CN(C)CCN1C Chemical compound c1c(OC)cccc1NC(=O)c1cc([N+]([O-])=O)ccc1\N=C1/CN(C)CCN1C CODDAXSDXHVTSW-ZBJSNUHESA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 description 1
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical compound [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- BPKIGYQJPYCAOW-FFJTTWKXSA-I calcium;potassium;disodium;(2s)-2-hydroxypropanoate;dichloride;dihydroxide;hydrate Chemical compound O.[OH-].[OH-].[Na+].[Na+].[Cl-].[Cl-].[K+].[Ca+2].C[C@H](O)C([O-])=O BPKIGYQJPYCAOW-FFJTTWKXSA-I 0.000 description 1
- 238000004422 calculation algorithm Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- ARYTXMNEANMLMU-ATEDBJNTSA-N campestanol Chemical compound C([C@@H]1CC2)[C@@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)CC[C@@H](C)C(C)C)[C@@]2(C)CC1 ARYTXMNEANMLMU-ATEDBJNTSA-N 0.000 description 1
- SGNBVLSWZMBQTH-PODYLUTMSA-N campesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](C)C(C)C)[C@@]1(C)CC2 SGNBVLSWZMBQTH-PODYLUTMSA-N 0.000 description 1
- 235000000431 campesterol Nutrition 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 125000005518 carboxamido group Chemical group 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229940105329 carboxymethylcellulose Drugs 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000008614 cellular interaction Effects 0.000 description 1
- 230000030570 cellular localization Effects 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- BHONFOAYRQZPKZ-LCLOTLQISA-N chembl269478 Chemical compound C([C@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O)C1=CC=CC=C1 BHONFOAYRQZPKZ-LCLOTLQISA-N 0.000 description 1
- OEPQJOFGXUCSSY-LYBHJNIJSA-N chembl3181976 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC(Cl)=C(C#N)C=C1C(=O)NC1=CC=CC=C1 OEPQJOFGXUCSSY-LYBHJNIJSA-N 0.000 description 1
- KKZUIWZVXFPMAU-DYTRJAOYSA-N chembl3182102 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC=C([N+]([O-])=O)C=C1C(=O)NC1=CC=CC=C1F KKZUIWZVXFPMAU-DYTRJAOYSA-N 0.000 description 1
- JJXBPMRSKCGVTH-RELWKKBWSA-N chembl3183397 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC=C(F)C=C1C(=O)NC1=CC=CC=C1 JJXBPMRSKCGVTH-RELWKKBWSA-N 0.000 description 1
- IQVANPTXQFZVGN-LYBHJNIJSA-N chembl3184472 Chemical compound C1N(C)CCN(C)\C1=N\C1=CC=C(C#N)C=C1C(=O)NC1=CC=CC(F)=C1 IQVANPTXQFZVGN-LYBHJNIJSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 125000002668 chloroacetyl group Chemical group ClCC(=O)* 0.000 description 1
- GGCLNOIGPMGLDB-GYKMGIIDSA-N cholest-5-en-3-one Chemical compound C1C=C2CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 GGCLNOIGPMGLDB-GYKMGIIDSA-N 0.000 description 1
- NYOXRYYXRWJDKP-UHFFFAOYSA-N cholestenone Natural products C1CC2=CC(=O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 NYOXRYYXRWJDKP-UHFFFAOYSA-N 0.000 description 1
- BHYOQNUELFTYRT-DPAQBDIFSA-N cholesterol sulfate Chemical compound C1C=C2C[C@@H](OS(O)(=O)=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 BHYOQNUELFTYRT-DPAQBDIFSA-N 0.000 description 1
- 235000019416 cholic acid Nutrition 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 229960002471 cholic acid Drugs 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000004154 complement system Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000002872 contrast media Substances 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 239000011258 core-shell material Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 229960005168 croscarmellose Drugs 0.000 description 1
- 229960000913 crospovidone Drugs 0.000 description 1
- 239000001767 crosslinked sodium carboxy methyl cellulose Substances 0.000 description 1
- 125000000000 cycloalkoxy group Chemical group 0.000 description 1
- 125000005112 cycloalkylalkoxy group Chemical group 0.000 description 1
- 125000001995 cyclobutyl group Chemical group [H]C1([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 1
- 125000000582 cycloheptyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 1
- 125000000596 cyclohexenyl group Chemical group C1(=CCCCC1)* 0.000 description 1
- 125000002433 cyclopentenyl group Chemical group C1(=CCCC1)* 0.000 description 1
- 125000001559 cyclopropyl group Chemical group [H]C1([H])C([H])([H])C1([H])* 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- FMGSKLZLMKYGDP-USOAJAOKSA-N dehydroepiandrosterone Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC=C21 FMGSKLZLMKYGDP-USOAJAOKSA-N 0.000 description 1
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 1
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 229940039227 diagnostic agent Drugs 0.000 description 1
- UMGXUWVIJIQANV-UHFFFAOYSA-M didecyl(dimethyl)azanium;bromide Chemical compound [Br-].CCCCCCCCCC[N+](C)(C)CCCCCCCCCC UMGXUWVIJIQANV-UHFFFAOYSA-M 0.000 description 1
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 125000004852 dihydrofuranyl group Chemical group O1C(CC=C1)* 0.000 description 1
- 125000001070 dihydroindolyl group Chemical group N1(CCC2=CC=CC=C12)* 0.000 description 1
- QBSJHOGDIUQWTH-UHFFFAOYSA-N dihydrolanosterol Natural products CC(C)CCCC(C)C1CCC2(C)C3=C(CCC12C)C4(C)CCC(C)(O)C(C)(C)C4CC3 QBSJHOGDIUQWTH-UHFFFAOYSA-N 0.000 description 1
- 125000005043 dihydropyranyl group Chemical group O1C(CCC=C1)* 0.000 description 1
- 125000005057 dihydrothienyl group Chemical group S1C(CC=C1)* 0.000 description 1
- OGQYPPBGSLZBEG-UHFFFAOYSA-N dimethyl(dioctadecyl)azanium Chemical compound CCCCCCCCCCCCCCCCCC[N+](C)(C)CCCCCCCCCCCCCCCCCC OGQYPPBGSLZBEG-UHFFFAOYSA-N 0.000 description 1
- WQLVFSAGQJTQCK-VKROHFNGSA-N diosgenin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)CC[C@H](O)CC4=CC[C@H]3[C@@H]2C1)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 WQLVFSAGQJTQCK-VKROHFNGSA-N 0.000 description 1
- WQLVFSAGQJTQCK-UHFFFAOYSA-N diosgenin Natural products CC1C(C2(CCC3C4(C)CCC(O)CC4=CCC3C2C2)C)C2OC11CCC(C)CO1 WQLVFSAGQJTQCK-UHFFFAOYSA-N 0.000 description 1
- USIUVYZYUHIAEV-UHFFFAOYSA-N diphenyl ether Chemical compound C=1C=CC=CC=1OC1=CC=CC=C1 USIUVYZYUHIAEV-UHFFFAOYSA-N 0.000 description 1
- 206010013023 diphtheria Diseases 0.000 description 1
- KPUWHANPEXNPJT-UHFFFAOYSA-N disiloxane Chemical compound [SiH3]O[SiH3] KPUWHANPEXNPJT-UHFFFAOYSA-N 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 125000005303 dithiazolyl group Chemical group S1SNC(=C1)* 0.000 description 1
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 235000012489 doughnuts Nutrition 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 239000003974 emollient agent Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000002121 endocytic effect Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 229940007078 entamoeba histolytica Drugs 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- JGENYNHRIOHZOP-UHFFFAOYSA-N ethyl 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCOP([O-])(=O)OCC[N+](C)(C)C JGENYNHRIOHZOP-UHFFFAOYSA-N 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 229940118764 francisella tularensis Drugs 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
- 125000002541 furyl group Chemical group 0.000 description 1
- 229940059947 gadolinium Drugs 0.000 description 1
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 108010055409 ganglioside receptor Proteins 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000012637 gene transfection Methods 0.000 description 1
- 229940085435 giardia lamblia Drugs 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- JEJLGIQLPYYGEE-UHFFFAOYSA-N glycerol dipalmitate Natural products CCCCCCCCCCCCCCCC(=O)OCC(CO)OC(=O)CCCCCCCCCCCCCCC JEJLGIQLPYYGEE-UHFFFAOYSA-N 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000021061 grooming behavior Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 125000004404 heteroalkyl group Chemical group 0.000 description 1
- 125000004474 heteroalkylene group Chemical group 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 238000013537 high throughput screening Methods 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 125000002632 imidazolidinyl group Chemical group 0.000 description 1
- 125000002636 imidazolinyl group Chemical group 0.000 description 1
- 125000002883 imidazolyl group Chemical group 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000126 in silico method Methods 0.000 description 1
- 229940055742 indium-111 Drugs 0.000 description 1
- APFVFJFRJDLVQX-AHCXROLUSA-N indium-111 Chemical compound [111In] APFVFJFRJDLVQX-AHCXROLUSA-N 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000000976 ink Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 125000002183 isoquinolinyl group Chemical group C1(=NC=CC2=CC=CC=C12)* 0.000 description 1
- 125000005956 isoquinolyl group Chemical group 0.000 description 1
- 125000004628 isothiazolidinyl group Chemical group S1N(CCC1)* 0.000 description 1
- 125000000842 isoxazolyl group Chemical group 0.000 description 1
- 125000000468 ketone group Chemical group 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 229940058690 lanosterol Drugs 0.000 description 1
- CAHGCLMLTWQZNJ-RGEKOYMOSA-N lanosterol Chemical compound C([C@]12C)C[C@@H](O)C(C)(C)[C@H]1CCC1=C2CC[C@]2(C)[C@H]([C@H](CCC=C(C)C)C)CC[C@@]21C CAHGCLMLTWQZNJ-RGEKOYMOSA-N 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 125000000400 lauroyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 235000005772 leucine Nutrition 0.000 description 1
- 125000002463 lignoceryl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000005647 linker group Chemical group 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 229960003151 mercaptamine Drugs 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 235000006109 methionine Nutrition 0.000 description 1
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 1
- 125000004184 methoxymethyl group Chemical group [H]C([H])([H])OC([H])([H])* 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 125000004170 methylsulfonyl group Chemical group [H]C([H])([H])S(*)(=O)=O 0.000 description 1
- 125000004092 methylthiomethyl group Chemical group [H]C([H])([H])SC([H])([H])* 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- KWXIRENOASTEIJ-UHFFFAOYSA-N molport-010-674-649 Chemical compound N12C=CC=C2C(C(C)C)N(C(=O)NCCCOCC)CC2=C1SC1=C2CCCC1 KWXIRENOASTEIJ-UHFFFAOYSA-N 0.000 description 1
- FPHACODWEVNLSA-UHFFFAOYSA-N molport-010-674-670 Chemical compound N12C=CC=C2C(CC)N(C(=O)NCCCOC)CC2=C1SC1=C2CCCC1 FPHACODWEVNLSA-UHFFFAOYSA-N 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 125000002950 monocyclic group Chemical group 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 125000002757 morpholinyl group Chemical group 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000002636 mycotoxin Substances 0.000 description 1
- 125000001421 myristyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- MCMDLYDRUNNJKR-UHFFFAOYSA-N n-(1,3-benzothiazol-2-yl)-n-methyl-2-thiophen-2-ylacetamide Chemical compound N=1C2=CC=CC=C2SC=1N(C)C(=O)CC1=CC=CS1 MCMDLYDRUNNJKR-UHFFFAOYSA-N 0.000 description 1
- PYFYONWWATZIHY-UHFFFAOYSA-N n-(4,5,6,7-tetrahydro-1,3-benzothiazol-2-yl)-2-thiophen-2-ylacetamide Chemical compound N=1C=2CCCCC=2SC=1NC(=O)CC1=CC=CS1 PYFYONWWATZIHY-UHFFFAOYSA-N 0.000 description 1
- GGTSRJJDPBAFFD-XLNRJJMWSA-N n-[(z)-1-[4-(dimethylamino)phenyl]-3-(3-imidazol-1-ylpropylamino)-3-oxoprop-1-en-2-yl]benzamide Chemical compound C1=CC(N(C)C)=CC=C1\C=C(C(=O)NCCCN1C=NC=C1)/NC(=O)C1=CC=CC=C1 GGTSRJJDPBAFFD-XLNRJJMWSA-N 0.000 description 1
- DAXGRRHAZMMKLL-UHFFFAOYSA-N n-[1-[2-[2-chloro-6-(4-chlorophenoxy)pyridin-4-yl]-4-methyl-1,3-thiazol-5-yl]ethylideneamino]aniline Chemical compound S1C(C=2C=C(OC=3C=CC(Cl)=CC=3)N=C(Cl)C=2)=NC(C)=C1C(C)=NNC1=CC=CC=C1 DAXGRRHAZMMKLL-UHFFFAOYSA-N 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- NMIZCAMXWFYFDO-UHFFFAOYSA-N n-phenyl-n-(4,5,6,7-tetrahydro-1,3-benzothiazol-2-yl)-2-thiophen-2-ylacetamide Chemical compound N=1C=2CCCCC=2SC=1N(C=1C=CC=CC=1)C(=O)CC1=CC=CS1 NMIZCAMXWFYFDO-UHFFFAOYSA-N 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 229940031182 nanoparticles iron oxide Drugs 0.000 description 1
- 125000001971 neopentyl group Chemical group [H]C([*])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000000696 nitrogen adsorption--desorption isotherm Methods 0.000 description 1
- 125000000018 nitroso group Chemical group N(=O)* 0.000 description 1
- 239000012454 non-polar solvent Substances 0.000 description 1
- 239000012457 nonaqueous media Substances 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 210000000633 nuclear envelope Anatomy 0.000 description 1
- 108700020942 nucleic acid binding protein Proteins 0.000 description 1
- 102000044158 nucleic acid binding protein Human genes 0.000 description 1
- 108060005597 nucleoplasmin Proteins 0.000 description 1
- 229920002113 octoxynol Polymers 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 125000001715 oxadiazolyl group Chemical group 0.000 description 1
- 125000000160 oxazolidinyl group Chemical group 0.000 description 1
- 125000002971 oxazolyl group Chemical group 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 108010043655 penetratin Proteins 0.000 description 1
- MCYTYTUNNNZWOK-LCLOTLQISA-N penetratin Chemical compound C([C@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(N)=O)C1=CC=CC=C1 MCYTYTUNNNZWOK-LCLOTLQISA-N 0.000 description 1
- 125000006340 pentafluoro ethyl group Chemical group FC(F)(F)C(F)(F)* 0.000 description 1
- 239000000863 peptide conjugate Substances 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 125000004193 piperazinyl group Chemical group 0.000 description 1
- 125000003386 piperidinyl group Chemical group 0.000 description 1
- 108010011110 polyarginine Proteins 0.000 description 1
- 229920002523 polyethylene Glycol 1000 Polymers 0.000 description 1
- 229940057847 polyethylene glycol 600 Drugs 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 235000013809 polyvinylpolypyrrolidone Nutrition 0.000 description 1
- 229920000523 polyvinylpolypyrrolidone Polymers 0.000 description 1
- 238000002459 porosimetry Methods 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 229940069328 povidone Drugs 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 229960000249 pregnenolone Drugs 0.000 description 1
- ORNBQBCIOKFOEO-QGVNFLHTSA-N pregnenolone Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 ORNBQBCIOKFOEO-QGVNFLHTSA-N 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- WGYKZJWCGVVSQN-UHFFFAOYSA-N propylamine Chemical group CCCN WGYKZJWCGVVSQN-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 231100000654 protein toxin Toxicity 0.000 description 1
- 125000004309 pyranyl group Chemical group O1C(C=CC=C1)* 0.000 description 1
- 125000003373 pyrazinyl group Chemical group 0.000 description 1
- 125000003072 pyrazolidinyl group Chemical group 0.000 description 1
- 125000002755 pyrazolinyl group Chemical group 0.000 description 1
- 125000003226 pyrazolyl group Chemical group 0.000 description 1
- 125000002098 pyridazinyl group Chemical group 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- 125000001422 pyrrolinyl group Chemical group 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 125000002943 quinolinyl group Chemical group N1=C(C=CC2=CC=CC=C12)* 0.000 description 1
- 125000005493 quinolyl group Chemical group 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 230000000979 retarding effect Effects 0.000 description 1
- 229940108325 retinyl palmitate Drugs 0.000 description 1
- 235000019172 retinyl palmitate Nutrition 0.000 description 1
- 239000011769 retinyl palmitate Substances 0.000 description 1
- KNUXHTWUIVMBBY-JRJYXWDASA-N rintatolimod Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(O)=O)O1.O[C@@H]1[C@H](O)[C@@H](COP(O)(O)=O)O[C@H]1N1C(=O)NC(=O)C=C1.O[C@@H]1[C@H](O)[C@@H](COP(O)(O)=O)O[C@H]1N1C(NC=NC2=O)=C2N=C1 KNUXHTWUIVMBBY-JRJYXWDASA-N 0.000 description 1
- 229950006564 rintatolimod Drugs 0.000 description 1
- 229930195734 saturated hydrocarbon Natural products 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 229960001153 serine Drugs 0.000 description 1
- 239000004208 shellac Substances 0.000 description 1
- ZLGIYFNHBLSMPS-ATJNOEHPSA-N shellac Chemical compound OCCCCCC(O)C(O)CCCCCCCC(O)=O.C1C23[C@H](C(O)=O)CCC2[C@](C)(CO)[C@@H]1C(C(O)=O)=C[C@@H]3O ZLGIYFNHBLSMPS-ATJNOEHPSA-N 0.000 description 1
- 229940113147 shellac Drugs 0.000 description 1
- 235000013874 shellac Nutrition 0.000 description 1
- 231100000161 signs of toxicity Toxicity 0.000 description 1
- 150000004756 silanes Chemical class 0.000 description 1
- 150000004819 silanols Chemical class 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 125000003808 silyl group Chemical group [H][Si]([H])([H])[*] 0.000 description 1
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 description 1
- 229950005143 sitosterol Drugs 0.000 description 1
- 235000015500 sitosterol Nutrition 0.000 description 1
- NLQLSVXGSXCXFE-UHFFFAOYSA-N sitosterol Natural products CC=C(/CCC(C)C1CC2C3=CCC4C(C)C(O)CCC4(C)C3CCC2(C)C1)C(C)C NLQLSVXGSXCXFE-UHFFFAOYSA-N 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 230000011273 social behavior Effects 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000008354 sodium chloride injection Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229960001790 sodium citrate Drugs 0.000 description 1
- 239000008109 sodium starch glycolate Substances 0.000 description 1
- 229920003109 sodium starch glycolate Polymers 0.000 description 1
- 229940079832 sodium starch glycolate Drugs 0.000 description 1
- 239000002594 sorbent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 210000004215 spore Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- LGJMUZUPVCAVPU-HRJGVYIJSA-N stigmastanol Chemical compound C([C@@H]1CC2)[C@@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]2(C)CC1 LGJMUZUPVCAVPU-HRJGVYIJSA-N 0.000 description 1
- 229940032091 stigmasterol Drugs 0.000 description 1
- HCXVJBMSMIARIN-PHZDYDNGSA-N stigmasterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)/C=C/[C@@H](CC)C(C)C)[C@@]1(C)CC2 HCXVJBMSMIARIN-PHZDYDNGSA-N 0.000 description 1
- 235000016831 stigmasterol Nutrition 0.000 description 1
- BFDNMXAIBMJLBB-UHFFFAOYSA-N stigmasterol Natural products CCC(C=CC(C)C1CCCC2C3CC=C4CC(O)CCC4(C)C3CCC12C)C(C)C BFDNMXAIBMJLBB-UHFFFAOYSA-N 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 125000006296 sulfonyl amino group Chemical group [H]N(*)S(*)(=O)=O 0.000 description 1
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 238000006557 surface reaction Methods 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 108010053343 temporin Proteins 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical group C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 125000006169 tetracyclic group Chemical group 0.000 description 1
- 125000003718 tetrahydrofuranyl group Chemical group 0.000 description 1
- VUYXVWGKCKTUMF-UHFFFAOYSA-N tetratriacontaethylene glycol monomethyl ether Chemical compound COCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO VUYXVWGKCKTUMF-UHFFFAOYSA-N 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000001113 thiadiazolyl group Chemical group 0.000 description 1
- 125000005458 thianyl group Chemical group 0.000 description 1
- 125000000335 thiazolyl group Chemical group 0.000 description 1
- 125000001544 thienyl group Chemical group 0.000 description 1
- 125000001583 thiepanyl group Chemical group 0.000 description 1
- 125000002053 thietanyl group Chemical group 0.000 description 1
- 125000001730 thiiranyl group Chemical group 0.000 description 1
- 125000004568 thiomorpholinyl group Chemical group 0.000 description 1
- 239000004408 titanium dioxide Substances 0.000 description 1
- 235000010215 titanium dioxide Nutrition 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 125000002088 tosyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1C([H])([H])[H])S(*)(=O)=O 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 108010062760 transportan Proteins 0.000 description 1
- 125000001425 triazolyl group Chemical group 0.000 description 1
- 125000006168 tricyclic group Chemical group 0.000 description 1
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 1
- 125000000026 trimethylsilyl group Chemical group [H]C([H])([H])[Si]([*])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 210000005166 vasculature Anatomy 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 244000052613 viral pathogen Species 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 210000002268 wool Anatomy 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- CGSJXLIKVBJVRY-XTGBIJOFSA-N zymosterol Chemical compound C([C@@]12C)C[C@H](O)C[C@@H]1CCC1=C2CC[C@]2(C)[C@@H]([C@@H](CCC=C(C)C)C)CC[C@H]21 CGSJXLIKVBJVRY-XTGBIJOFSA-N 0.000 description 1
- AUNLIRXIJAVBNM-ZSBATXSLSA-N zymosterol intermediate 2 Chemical compound C([C@@]12C)CC(=O)C[C@@H]1CCC1=C2CC[C@]2(C)[C@@H]([C@@H](CCC=C(C)C)C)CC[C@H]21 AUNLIRXIJAVBNM-ZSBATXSLSA-N 0.000 description 1
Images
Classifications
-
- 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
- A61K47/6905—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 colloid or an emulsion
- A61K47/6911—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 colloid or an emulsion the form being a liposome
- A61K47/6915—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 colloid or an emulsion the form being a liposome the form being a liposome with polymerisable or polymerized bilayer-forming substances, e.g. polymersomes
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/4965—Non-condensed pyrazines
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
-
- 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
- A61K47/54—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 compound
- A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
-
- 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
- A61K47/6905—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 colloid or an emulsion
- A61K47/6911—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 colloid or an emulsion the form being a liposome
- A61K47/6913—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 colloid or an emulsion the form being a liposome the liposome being modified on its surface by an antibody
-
- 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
- A61K47/6905—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 colloid or an emulsion
- A61K47/6917—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 colloid or an emulsion the form being a lipoprotein vesicle, e.g. HDL or LDL proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5123—Organic compounds, e.g. fats, sugars
Definitions
- a sequence listing appendix including an ASCII formatted file accompanies this application.
- the appendix includes a file named “SD14659.2 DIV_ST25.txt,” created on Apr. 14, 2021 (size of 12.7 kilobytes), which is hereby incorporated by reference in its entirety.
- the present invention relates to lipid-coated particles for treating viral infections, including viral encephalitis infections.
- an antiviral compound can be disposed within the lipid-coated particle, thereby providing an antiviral carrier. Methods of making and using such carriers are described herein.
- Promising therapeutics can often provide beneficial in vitro characteristics, yet fail during in vivo investigations. Various difficulties can arise, including adverse effects at therapeutic doses, limited bioavailability, and chemical instability. Accordingly, there is need for additional methodologies to improve delivery of such therapeutics to the desired site of action.
- the present invention relates to the use of a lipid-coated particle to improve the solubility and/or stability of a drug (e.g., an antiviral drug).
- a drug e.g., an antiviral drug
- lipid-coated mesoporous silica nanoparticles were employed as delivery vehicles for antivirals with known solubility and stability issues.
- the large surface area of the MSN core likely promotes loading of a hydrophobic or lipophilic drug, while the liposome coating could enable enhanced circulation time and biocompatibility, thereby providing an ideal carrier for antiviral ML336 drug delivery.
- LC-MSNs Provides are determinations regarding colloidal stability; in vitro viral inhibition in a dose-dependent manner, as compared to untreated controls; and in vivo studies related to toxicity and efficacy in reducing brain viral titer of a virus (e.g., Venezuelan equine encephalitis virus (VEEV) TC-83 in mice).
- VEEV Venezuelan equine encephalitis virus
- the present invention features a method of increasing a stability and/or a solubility of a compound within an aqueous solution.
- the method includes: incubating the compound with a core comprising a plurality of pores, thereby providing a loaded core; and coating the loaded core with a lipid layer, thereby provided a lipid-coated particle, wherein the stability and/or the solubility of the lipid-coated particle within the aqueous solution is greater than the stability and/or the solubility of the compound within the aqueous solution.
- the compound has an aqueous solubility of from about 20 ⁇ g/mL to about 150 ⁇ g/mL in phosphate-buffered saline at a pH of 7.4 and/or a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- the present invention features a method of treating a viral infection.
- the method includes administering an effective amount of a lipid-coated particle to a subject.
- the lipid-coated particle includes a porous core, a lipid layer, and an antiviral compound disposed within at least one pore of the porous core.
- a concentration of the effective amount of the antiviral compound within the lipid-coated particle is less than a concentration of the effective amount of the antiviral compound alone.
- the viral infection is an alphavirus infection. In other embodiments, the viral infection is an encephalitis infection.
- the present invention features a method of treating viral encephalitis.
- the method includes administering an effective amount of a lipid-coated particle to a subject.
- the lipid-coated particle includes a porous core and an antiviral compound disposed within at least one pore of the porous core, where the effective amount of the lipid-coated particle provides a reduction of brain viral load, as compared to administration of the antiviral compound alone.
- the present invention features a method of reducing brain viral load within a subject.
- the method includes administering an effective amount of a lipid-coated particle to the subject.
- the lipid-coated particle comprises a porous core and an antiviral compound disposed within at least one pore of the porous core.
- the present invention features an antiviral carrier including: a porous core comprising a plurality of pores; an antiviral compound disposed in at least one pore; and a lipid layer disposed around the porous core.
- the antiviral compound has an aqueous solubility of from about 20 ⁇ g/mL to about 150 ⁇ g/mL in phosphate-buffered saline at a pH of 7.4 and/or a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- the antiviral compound is present in an amount of from about 10 ⁇ g/mg to 50 ⁇ g/mg ( ⁇ g of the compound per mg of the carrier).
- the antiviral compound has a release rate of from about 3 ⁇ g/mg to about 20 ⁇ g/mg ( ⁇ g of the compound per mg of the carrier) over a period of about 24 hours in vitro.
- the lipid layer includes a zwitterionic lipid, a cholesterol or a derivative thereof, and a pegylated lipid.
- the antiviral compound has an aqueous solubility of from about 20 ⁇ g/mL to about 150 ⁇ g/mL in phosphate-buffered saline at a pH of 7.4. In other embodiments, the antiviral compound has a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- the antiviral compound has an EC 50 value of from about 0.01 ⁇ M to about 1 ⁇ M as determined in a cellular assay.
- the antiviral compound has an EC 90 value of from about 100 nM to about 300 nM as determined in a cellular assay. In other embodiments, the antiviral compound is hydrophobic or lipophilic.
- the present invention features a formulation including an antiviral (e.g., any described herein) and an optional pharmaceutically acceptable excipient.
- the lipid-coated particle includes a porous core and an antiviral compound disposed within at least one pore of the porous core.
- the lipid layer includes a zwitterionic lipid, a cholesterol or a derivative thereof, and a pegylated lipid.
- the compound or the antiviral compound has an aqueous solubility of from about 20 ⁇ g/mL to about 150 ⁇ g/mL in phosphate-buffered saline at a pH of 7.4. In other embodiments, the compound or the antiviral compound has a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- the compound or the antiviral compound has an EC 50 value of from about 0.01 ⁇ M to about 1 ⁇ M as determined in a cellular assay. In other embodiments, the compound or the antiviral compound has an EC 90 value of from about 100 nM to about 300 nM as determined in a cellular assay.
- the compound or the antiviral compound is hydrophobic or lipophilic.
- the compound or the antiviral compound has a structure of formula (I) or (II) or (III):
- each R 2 is, independently, optionally substituted aryl (e.g., C 4-18 aryl, including optionally substituted phenyl, such as a substituted p-phenyl), optionally substituted heterocyclyl, optionally substituted alkaryl (e.g., C 1-6 alk-C 4-18 aryl, including optionally substituted benzyl), or optionally substituted alkheterocyclyl; each R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8 is, independently, H, optionally substituted alkyl (e.g., C 1-6 alkyl), halo, nitro, nitroso, amino, azido, carboxyl, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl, or in which R 4 and R 5
- each R 4 , R 5 , R 7 , and R 8 is, independently, H, optionally substituted alkyl, halo, nitro, amino, azido, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl, or in which R 4 and R 5 , taken together, or R 7 and R 8 , taken together, form an optionally substituted spirocyclyl; and each R 3 and R 6 is, independently, H, optionally substituted alkyl, halo, nitro, nitroso, amino, azido, carboxyl, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl.
- the term “about” means +/ ⁇ 10% of any recited value. As used herein, this term modifies any recited value, range of values, or endpoints of one or more ranges.
- alkaryl is meant an aryl group, as defined herein, attached to the parent molecular group through an alkylene group, as defined herein.
- alkheteroaryl is meant a heteroaryl group, as defined herein, attached to the parent molecular group through an alkylene group.
- alk- Other groups preceded by the prefix “alk-” are defined in the same manner.
- the alkaryl group can be substituted or unsubstituted.
- the alkaryl group can be substituted with one or more substitution groups, as described herein for alkyl and/or aryl.
- Exemplary unsubstituted alkaryl groups are of from 7 to 16 carbons (C 7-16 alkaryl), as well as those having an alkylene group with 1 to 6 carbons and an aryl group with 4 to 18 carbons (i.e., C 1-6 alk-C 4-18 aryl).
- alkheterocyclyl represents a heterocyclyl group, as defined herein, attached to the parent molecular group through an alkylene group, as defined herein.
- exemplary unsubstituted alkheterocyclyl groups are of from 2 to 14 carbons.
- alkyl and the prefix “alk” is meant a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like.
- the alkyl group can be cyclic (e.g., C 3-24 cycloalkyl) or acyclic.
- the alkyl group can be branched or unbranched.
- the alkyl group can also be substituted or unsubstituted.
- the alkyl group can be substituted with one, two, three or, in the case of alkyl groups of two carbons or more, four substituents independently selected from the group consisting of: (1) C 1-6 alkoxy (e.g., —OAk, in which Ak is an alkyl group, as defined herein); (2) C 1-6 alkylsulfinyl (e.g., —S(O)Ak, in which Ak is an alkyl group, as defined herein); (3) C 1-6 alkylsulfonyl (e.g., —SO 2 Ak, in which Ak is an alkyl group, as defined herein); (4) amino (e.g., —NR N1 R N2 , where each of R N1 and R N2 is, independently, H or optionally substituted alkyl, or R N1 and R N2 , taken together with the nitrogen atom to which each are attached, form a heterocyclyl group); (5) aryl; (6) arylalkoxy (e.g
- the alkyl group can be a primary, secondary, or tertiary alkyl group substituted with one or more substituents (e.g., one or more halo or alkoxy).
- the unsubstituted alkyl group is a C 1-3 , C 1-6 , C 1-12 , C 1-16 , C 1-18 , C 1-20 , or C 1-24 alkyl group.
- alkylene is meant a multivalent (e.g., bivalent, trivalent, tetravalent, etc.) form of an alkyl group, as described herein.
- exemplary alkylene groups include methylene, ethylene, propylene, butylene, etc.
- the alkylene group is a C 1-3 , C 1-6 , C 1-12 , C 1-16 , C 1-18 , C 1-20 , C 1-24 , C 2-3 , C 2-6 , C 2-12 , C 2-16 , C 2-18 , C 2-20 , or C 2-24 alkylene group.
- the alkylene group can be branched or unbranched.
- the alkylene group can also be substituted or unsubstituted.
- the alkylene group can be substituted with one or more substitution groups, as described herein for alkyl.
- amino is meant —NR N1 R N2 , where each of R N1 and R N2 is, independently, H or optionally substituted alkyl, or R N1 and R N2 , taken together with the nitrogen atom to which each are attached, form a heterocyclyl group, as defined herein.
- aryl is meant a group that contains any carbon-based aromatic group including, but not limited to, benzyl, naphthalene, phenyl, biphenyl, phenoxybenzene, and the like.
- aryl also includes “heteroaryl,” which is defined as a group that contains an aromatic group that has at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus.
- non-heteroaryl which is also included in the term “aryl,” defines a group that contains an aromatic group that does not contain a heteroatom. The aryl group can be substituted or unsubstituted.
- the aryl group can be substituted with one, two, three, four, or five substituents independently selected from the group consisting of: (1) C 1-6 alkanoyl (e.g., —C(O)Ak, in which Ak is an alkyl group, as defined herein); (2) C 1-6 alkyl; (3) C 1-6 alkoxy (e.g., —OAk, in which Ak is an alkyl group, as defined herein); (4) C 1-6 alkoxy-C 1-6 alkyl (e.g., an alkyl group, which is substituted with an alkoxy group —OAk, in which Ak is an alkyl group, as defined herein); (5) C 1-6 alkylsulfinyl (e.g., —S(O)Ak, in which Ak is an alkyl group, as defined herein); (6) C 1-6 alkylsulfinyl-C 1-6 alkyl (e.g., an alkyl group, which is substituted by an alky
- azido is meant an —N 3 group.
- carboxyl is meant a —CO 2 H group.
- cyano is meant a —CN group.
- cycloalkyl is meant a monovalent saturated or unsaturated non-aromatic cyclic hydrocarbon group of from three to eight carbons, unless otherwise specified, and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[2.2.1.]heptyl and the like.
- the cycloalkyl group can also be substituted or unsubstituted.
- the cycloalkyl group can be substituted with one or more groups including those described herein for alkyl.
- halo is meant F, Cl, Br, or I.
- haloalkyl is meant an alkyl group, as defined herein, substituted with one or more halo.
- heteroalkyl an alkyl group, as defined herein, containing one, two, three, or four non-carbon heteroatoms (e.g., independently selected from the group consisting of nitrogen, oxygen, phosphorous, sulfur, or halo).
- heteroaryl is meant a subset of heterocyclyl groups, as defined herein, which are aromatic, i.e., they contain 4n+2 pi electrons within the mono- or multicyclic ring system.
- heterocyclyl is meant a 5-, 6- or 7-membered ring, unless otherwise specified, containing one, two, three, or four non-carbon heteroatoms (e.g., independently selected from the group consisting of nitrogen, oxygen, phosphorous, sulfur, or halo).
- the 5-membered ring has zero to two double bonds and the 6- and 7-membered rings have zero to three double bonds.
- heterocyclyl also includes bicyclic, tricyclic and tetracyclic groups in which any of the above heterocyclic rings is fused to one, two, or three rings independently selected from the group consisting of an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, and another monocyclic heterocyclic ring, such as indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, benzofuryl, benzothienyl and the like.
- Heterocyclics include thiiranyl, thietanyl, tetrahydrothienyl, thianyl, thiepanyl, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, pyrrolyl, pyrrolinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, homopiperidinyl, pyrazinyl, piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidiniyl, morpholinyl, thiomorpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidin
- hydroxyl is meant —OH.
- hydroxyalkyl is meant an alkyl group, as defined herein, substituted by one to three hydroxyl groups, with the proviso that no more than one hydroxyl group may be attached to a single carbon atom of the alkyl group and is exemplified by hydroxymethyl, dihydroxypropyl, and the like.
- nitro is meant an —NO 2 group.
- nitroalkyl is meant an alkyl group, as defined herein, substituted by one to three nitro groups.
- nitroso is meant an —NO group.
- perfluoroalkyl is meant an alkyl group, as defined herein, having each hydrogen atom substituted with a fluorine atom.
- exemplary perfluoroalkyl groups include trifluoromethyl, pentafluoroethyl, etc.
- protecting group is meant any group intended to protect a reactive group against undesirable synthetic reactions. Commonly used protecting groups are disclosed in “Greene's Protective Groups in Organic Synthesis,” John Wiley & Sons, New York, 2007 (4th ed., eds. P. G. M. Wuts and T. W. Greene), which is incorporated herein by reference.
- O-protecting groups include an optionally substituted alkyl group (e.g., forming an ether with reactive group O), such as methyl, methoxymethyl, methylthiomethyl, benzoyloxymethyl, t-butoxymethyl, etc.; an optionally substituted alkanoyl group (e.g., forming an ester with the reactive group O), such as formyl, acetyl, chloroacetyl, fluoroacetyl (e.g., perfluoroacetyl), methoxyacetyl, pivaloyl, t-butylacetyl, phenoxyacetyl, etc.; an optionally substituted aryloyl group (e.g., forming an ester with the reactive group O), such as —C(O)—Ar, including benzoyl; an optionally substituted alkylsulfonyl group (e.g., forming an alkylsulfonate with reactive group O), such as
- N-protecting groups include, e.g., formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, Boc, and Cbz.
- Such protecting groups can employ any useful agent to cleave the protecting group, thereby restoring the reactivity of the unprotected reactive group.
- salt is meant an ionic form of a compound or structure (e.g., any formulas, compounds, or compositions described herein), which includes a cation or anion compound to form an electrically neutral compound or structure.
- Salts e.g., simple salts having binary compounds, double salts, triple salts, etc.
- salts are well known in the art. For example, salts are described in Berge S M et al., “Pharmaceutical salts,” J. Pharm. Sci. 1977 January; 66(1):1-19; International Union of Pure and Applied Chemistry, “Nomenclature of Inorganic Chemistry,” Butterworth & Co.
- the salts can be prepared in situ during the final isolation and purification of the compounds of the invention or separately by reacting the free base group with a suitable organic acid (thereby producing an anionic salt) or by reacting the acid group with a suitable metal or organic salt (thereby producing a cationic salt).
- anionic salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, camphorate, camphorsulfonate, chloride, citrate, cyclopentanepropionate, digluconate, dihydrochloride, diphosphate, dodecylsulfate, edetate, ethanesulfonate, fumarate, glucoheptonate, glucomate, glutamate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, hydroxyethanesulfonate, hydroxynaphthoate, iodide, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malonate
- cationic salts include metal salts, such as alkali or alkaline earth salts, e.g., barium, calcium (e.g., calcium edetate), lithium, magnesium, potassium, sodium, and the like; other metal salts, such as aluminum, bismuth, iron, and zinc; as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, pyridinium, and the like.
- Other cationic salts include organic salts, such as chloroprocaine, choline, dibenzylethylenediamine, diethanolamine, ethylenediamine, methylglucamine, and procaine.
- spirocyclyl is meant an alkylene diradical, both ends of which are bonded to the same carbon atom of the parent group to form a spirocyclyl group and also a heteroalkylene diradical, both ends of which are bonded to the same atom.
- micro is meant having at least one dimension that is less than 1 mm.
- a microstructure e.g., any structure described herein, such as a microparticle
- nano is meant having at least one dimension that is less than 1 ⁇ m.
- a nanostructure e.g., any structure described herein, such as a nanoparticle
- phrases “effective average particle size” as used herein to describe a multiparticulate means that at least 50% of the particles therein are of a specified size. Accordingly, “effective average particle size of less than about 2,000 nm in diameter” means that at least 50% of the particles therein are less than about 2,000 nm in diameter.
- nanoparticulates have an effective average particle size of less than about 2,000 nm (i.e., 2 microns), less than about 1,900 nm, less than about 1,800 nm, less than about 1,700 nm, less than about 1,600 nm, less than about 1,500 nm, less than about 1,400 nm, less than about 1,300 nm, less than about 1,200 nm, less than about 1,100 nm, less than about 1,000 nm, less than about 900 nm, less than about 800 nm, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 250 nm, less than about 200 nm, less than about 150 nm, less than about 100 nm, less than about 75 nm, or less than about 50 nm, as measured by light-scattering methods, microscopy, or other appropriate methods.
- the particles are monodisperse and generally no greater than about 50 nm in average diameter, often less than about 30 nm in average diameter, as otherwise described herein.
- the term “D 50 ” refers to the particle size below which 50% of the particles in a multiparticulate fall.
- D 90 refers to the particle size below which 90% of the particles in a multiparticulate fall.
- the term “monodisperse” is used as a standard definition established by the National Institute of Standards and Technology (NIST) ( Particle Size Characterization, Special Publication 960-1, January 2001) to describe a distribution of particle size within a population of particles, in this case nanoparticles, which particle distribution may be considered monodisperse if at least 90% of the distribution lies within 5% of the median size. See, e.g., Takeuchi S et al., Adv. Mater. 2005; 17(8): 1067-72.
- lipid is used to describe the components which are used to form lipid mono-, bi-, or multilayers on the surface of the particles (e.g., a core of the particle), that are used in the present invention (e.g., as lipid-coated particles) and may include a PEGylated lipid.
- Various embodiments provide nanostructures, that are constructed from nanoparticles, which support one or more lipid layers (e.g., bilayer(s) or multilayer(s)).
- the nanostructures preferably include, for example, a core-shell structure including a porous particle core surrounded by a shell of one or more lipid bilayer(s).
- the nanostructure e.g., a porous silica or alum nanostructure
- targeting ligand and “targeting active species” are used to describe a compound or moiety (e.g., an antigen), which is complexed or covalently bonded to the surface of particle according to the present invention (e.g., either directly on an outer surface of a delivery platform, on an outer lipid layer, or on a supported lipid layer).
- the targeting ligand in turn, binds to a moiety on the surface of a cell to be targeted so that the lipid-coated particles may bind to the surface of the targeted cell, enter the cell or an organelle thereof, and/or deposit their contents into the cell.
- the targeting active species for use in the present invention is preferably a targeting peptide (e.g., a cell penetration peptide, a fusogenic peptide, or an endosomolytic peptide, as otherwise described herein), a polypeptide including an antibody or antibody fragment, an aptamer, or a carbohydrate, among other species that bind to a targeted cell.
- a targeting peptide e.g., a cell penetration peptide, a fusogenic peptide, or an endosomolytic peptide, as otherwise described herein
- a polypeptide including an antibody or antibody fragment, an aptamer, or a carbohydrate, among other species that bind to a targeted cell.
- an “effective amount” or a “sufficient amount” of an agent is that amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, an “effective amount” depends upon the context in which it is being applied.
- an agent e.g., a lipid-coated particle, an antiviral compound, a compound, or an antiviral carrier
- an “effective amount” depends upon the context in which it is being applied.
- an agent that employs an antiviral compound is, for example, an amount sufficient to achieve decreased viral titer of a virus and/or to treat an infection, as compared to the response obtained without administration of the agent.
- an antiviral carrier that employs an antiviral compound in the context of administering an antiviral carrier that employs an antiviral compound is, for example, an amount sufficient to achieve decreased viral titer of a virus and/or to treat an infection, as compared to the response obtained without administration of the antiviral carrier.
- an antiviral carrier that employs an amount of the antiviral compound in the context of administering an antiviral carrier that employs an amount of the antiviral compound is, for example, an amount sufficient to achieve decreased viral titer of a virus and/or to treat an infection, as compared to the response obtained with administration of the amount of the antiviral compound without the antiviral carrier.
- an effective amount of an antiviral carrier including an antiviral compound can be compared to any useful control (e.g., an effect determined upon administration of an effective amount of the antiviral compound when used alone, an effect determined upon administration of a buffer, or an effect determined without administration of the antiviral carrier).
- subject is meant a human or non-human animal (e.g., a mammal).
- treating a disease, disorder, or condition in a subject is meant reducing at least one symptom of the disease, disorder, or condition by administrating a therapeutic agent to the subject.
- treating prophylactically a disease, disorder, or condition in a subject is meant reducing the frequency of occurrence of or reducing the severity of a disease, disorder or condition by administering a therapeutic agent to the subject prior to the onset of disease symptoms.
- Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; diminishment of extent of disease, disorder, or condition; stabilized (i.e., not worsening) state of disease, disorder, or condition; preventing spread of disease, disorder, or condition; delay or slowing the progress of the disease, disorder, or condition; amelioration or palliation of the disease, disorder, or condition; and remission (whether partial or total), whether detectable or undetectable.
- salt is meant an ionic form of a compound or structure (e.g., any formulas, compounds, or compositions described herein), which includes a cation or anion compound to form an electrically neutral compound or structure.
- Salts are well known in the art. For example, non-toxic salts are described in Berge S M et al., “Pharmaceutical salts,” J. Pharm. Sci. 1977 January; 66(1):1-19; and in “Handbook of Pharmaceutical Salts: Properties, Selection, and Use,” Wiley-VCH, April 2011 (2nd rev. ed., eds. P. H. Stahl and C. G. Wermuth).
- the salts can be prepared in situ during the final isolation and purification of the compounds of the invention or separately by reacting the free base group with a suitable organic acid (thereby producing an anionic salt) or by reacting the acid group with a suitable metal or organic salt (thereby producing a cationic salt).
- anionic salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, camphorate, camphorsulfonate, chloride, citrate, cyclopentanepropionate, digluconate, dihydrochloride, diphosphate, dodecyl sulfate, edetate, ethanesulfonate, fumarate, glucoheptonate, glucomate, glutamate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, hydroxyethanesulfonate, hydroxynaphthoate, iodide, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malon
- Representative cationic salts include metal salts, such as alkali or alkaline earth salts, e.g., barium, calcium (e.g., calcium edetate), lithium, magnesium, potassium, sodium, and the like; other metal salts, such as aluminum, bismuth, iron, and zinc; as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, pyridinium, and the like.
- metal salts such as alkali or alkaline earth salts, e.g., barium, calcium (e.g., calcium edetate), lithium, magnesium, potassium, sodium, and the like
- other metal salts such as aluminum, bismuth, iron, and zinc
- cationic salts include organic salts, such as chloroprocaine, choline, dibenzylethylenediamine, diethanolamine, ethylenediamine, methylglucamine, and procaine.
- exemplary salts include pharmaceutically acceptable salts.
- pharmaceutically acceptable salt is meant a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio.
- pharmaceutically acceptable excipient any ingredient other than a compound or structure (e.g., any formulas, compounds, or compositions described herein) and having the properties of being nontoxic and non-inflammatory in a subject.
- exemplary, non-limiting excipients include adjuvants, antiadherents, antioxidants, binders, carriers, coatings, compression aids, diluents, disintegrants, dispersing agents, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), isotonic carriers, lubricants, preservatives, printing inks, solvents, sorbents, stabilizers, suspensing or dispersing agents, surfactants, sweeteners, waters of hydration, or wetting agents.
- excipients can be selected from those approved, for example, by the United States Food and Drug Administration or other governmental agency as being acceptable for use in humans or domestic animals.
- exemplary excipients include, but are not limited to alcohol, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, glycerol, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactated Ringer's solution, lactose, magnesium stearate, maltitol, maltose, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben,
- top As used herein, the terms “top,” “bottom,” “upper,” “lower,” “above,” and “below” are used to provide a relative relationship between structures. The use of these terms does not indicate or require that a particular structure must be located at a particular location in the apparatus.
- FIG. 1A-1C shows schematics of exemplary particles.
- A particular components of an exemplary lipid-coated particle 110 , including an exemplary core 101 and an exemplary lipid layer 102 .
- B a schematic of another exemplary lipid-coated particle 1100 , including an exemplary core 1001 and an exemplary lipid layer 1002 .
- C a schematic for an exemplary loaded, lipid coated-mesoporous silica nanoparticle (LC-MSN), including exemplary components.
- LC-MSN lipid coated-mesoporous silica nanoparticle
- FIG. 2A-2B shows (A) a schematic of ML336 loaded LC-MSN fabrication.
- the antiviral ML336 was incubated overnight at 4° C. with MSNs at a 10% mass ratio.
- Liposomes prepared by combining 77.5% DSPC:2.5% DSPEPEG200:20% cholesterol at mole ratios, were fused to ML336 loaded MSN cores under bath sonication at a 5:1 mass ratio of liposomes:nanoparticles to form ML336 loaded LC-MSNs.
- a schematic (B) of formula (I) and (II) for an exemplary class of quinazolinone-based inhibitors are also provided.
- FIG. 3A-3I shows characterization of ML336 LC-MSNs.
- A TEM images and
- cryo-EM images of (C) ML336 loaded LC-MSNs and (D) unloaded LC-MSNs (scale bar 50 nm). Gray arrows point out examples of the lipid bilayer.
- (G) hexagonal structure is highlighted by the honey comb-like arrangement (A 1 ) and tubular channels (A 2 ).
- Axis (z) is parallel to the pore's plan.
- (H) a tilted image shows tubular channels in a hexagonal arrangement ending by cargo-accessible openings (pore mouth).
- FIG. 4A-4F shows (A) DLS measurements of ML336 loaded MSNs and loaded LC-MSNs over the course of a week.
- ML336 loading and release was determined by comparing sample absorbance values at 320 nm to a standard curve in (B) a lipid solution or (C) a PBS solution.
- FIG. 5A-5D shows that ML336 inhibits TC-83 and VEEV (ZPC738 virulent strain) in a dose-dependent manner.
- FIG. 6 shows that ML336 loaded LC-MSNs do not visibly affect cell viability.
- FIG. 7 shows that ML336 loaded LC-MSNs inhibit virus in a dose-dependent manner.
- FIG. 8A-8C shows that ML336 loaded LC-MSNs inhibited virus in vitro.
- A phase microscopy images of cells 24, 48, and 72 hours post infection. Noticeable cell death is observed in the unloaded LC-MSN and no treatment groups, as compared to loaded LC-MSN treatment group; and
- FIG. 9A-9D shows repeatability studies for LC-MSN viral inhibition in vitro.
- (A) is also depicted in FIG. 8C .
- FIG. 10A-10D shows LC-MSN cellular internalization by clathrin-mediated endocytosis.
- B quantified through flow cytometry.
- the inhibitor panel included those targeting pH dependent endocytosis (BAF), clathrin-mediated endocytosis (DYN, CPZ), macropinocytosis (WORT, IPA-3), and caveola-mediated endocytosis (PMA, DYN), while untreated cells with (NI) and without LC-MSN (cells) addition served as controls.
- FIG. 11A-11C shows that inhibitors of clathrin-mediated endocytosis reduce both LC-MSN uptake and VSV infection, while inhibitors of caveolae-mediated endocytosis block RVFV and some VSV infection, but not LC-MSN cellular internalization.
- the inhibitor panel included those targeting pH dependent endocytosis (BAF), clathrin-mediated endocytosis (DYN, CPZ), macropinocytosis (WORT, IPA-3) and caveola-mediated endocytosis (PMA, DYN), while untreated cells with (NI) and without LC-MSN (cells) addition served as controls.
- FIG. 12A-12C shows unloaded LC-MSNs do not affect animal weight in safety studies.
- A Percent weight change in animals dosed with unloaded LC-MSNs or PBS alone over the course of 15 days.
- B Normalized weights of lung, liver, spleen, kidney, and brain to total animal weight in animals dosed with unloaded LC-MSNs or PBS alone over the course of 15 days (data are depicted as mean ⁇ standard deviation).
- C histological analysis of LC-MSN dosed C3H/HeN mice.
- Mouse tissues were dissected and formalin-fixed on day 15 post-treatment with a vehicle control (PBS) or LC-MSNs at 0.11 g LC-MSNs/kg/day for four days. Histological specimens were prepared through paraffin embedding and sectioning, followed by hematoxylin and eosin staining. Three animals per group were analyzed and representative images are shown. In the brain, the outer cortex is shown and displayed no obvious differences between the LC-MSN dosed and control groups. Similarly, the spleen and kidney sections exhibited normal morphology without signs of toxicity. In some samples, granulomas that contained collections of macrophages embedded in the lung and liver (indicated by asterisks) were identified and indicative of very mild symptoms.
- PBS vehicle control
- LC-MSNs LC-MSNs at 0.11 g LC-MSNs/kg/day for four days. Histological specimens were prepared through paraffin embedding and sectioning, followed by hematoxy
- FIG. 13A-13C shows ML336 loaded LC-MSNs show reduction of viral load in vivo.
- A a survival curve for TC-83 infected animals treated with 1 mg ML336 loaded LC-MSNs twice a day for 4 days, as well as viral load in
- B brain
- FIG. 14A-14C shows viral titer in tissues of TC-83 infected mice.
- the viral loads in (A) serum, (B) kidney, and (C) liver at day 4 post-infection via intranasal challenge of C3H/HeN mice with VEEV strain TC-83 were measured by standard plaque assays normalized to volume (ml) or organs mass (gram).
- Viral loads from four treatment conditions are shown for ML-336 loaded LC-MSN (circle), unloaded LC-MSN (square), free ML-336 (triangle), and vehicle only (PBS) (upside down triangle) with mean from 5 samples per condition.
- the limit of detection (LOD) is 100PFU, and samples at or below this threshold are all listed at LOD.
- the present invention relates to the use of lipid-coated particles to deliver compounds having reduced stability and/or solubility.
- ML336 is a small molecule inhibitor that displays antiviral activity but has poor stability and solubility characteristics.
- LC-MSNs lipid coated mesoporous silica nanoparticles
- VEEV Venezuelan equine encephalitis virus
- ML336-loaded LC-MSNs showed significant reduction in brain viral titer in VEEV TC-83 infected mice, as compared to PBS treated controls. Overall, these results highlight the utility of LC-MSNs as drug delivery vehicles to treat VEEV infections.
- FIG. 1A provides an exemplary lipid-coated particle 110 including an inner core 101 and an outer lipid layer 102 (e.g., a lipid bilayer, a multilamellar lipid layer, etc.) disposed around the core 101 .
- the core is porous (e.g., including a plurality of cores).
- the lipid layer can include any useful lipid (e.g., a PEGylated lipid), useful component (e.g., a cholesterol), and/or useful targeting ligand (e.g., any described herein).
- the outer lipid layer can include a plurality of layers, in which each layer can be a lipid bilayer. In this manner, the layer can be multilamellar because it includes multiple lamellae (or multiple layers).
- FIG. 1B provided another an exemplary lipid-coated particle 1100 including an inner core 1001 and an outer lipid layer 1002 (e.g., a lipid bilayer, a multilamellar lipid layer, etc.) disposed around the core 1001 .
- the core can have any useful features or characteristics.
- the core is a monosized (e.g., polydispersity index ⁇ 0.1) particle.
- the core can have any useful shape, morphology, pore size, and pore distribution.
- the lipid-coated particle can be characterized by any useful manner either before loading of cargo or after loading of cargo (e.g., overall charge, dimension, dispersity, etc.).
- components of the particle e.g., the core or the lipid layer
- can also be characterized by any useful manner e.g., pore size, core size, core charge, lipid layer thickness, lipid layer charge, etc.).
- Cargo e.g., a compound
- Cargo can be loaded in any useful manner.
- cargo is introduced to the core, and then the loaded core is exposed to a solution containing liposomes, which results in the formation of a lipid layer disposed around the loaded core.
- the present invention can include the any useful compound (e.g., an antiviral compound).
- the compound has reduced stability and/or reduced solubility, thereby would benefit from the use of a carrier (e.g., any described herein).
- the compound is hydrophobic (e.g., determined in any useful manner, such as any herein).
- the compound has limited aqueous solubility (e.g., from about 20 ⁇ g/mL to about 150 ⁇ g/mL, such as from 20 ⁇ g/mL to 50 ⁇ g/mL, 20 ⁇ g/mL to 100 ⁇ g/mL, 20 ⁇ g/mL to 150 ⁇ g/mL, 50 ⁇ g/mL to 100 ⁇ g/mL, or 50 ⁇ g/mL to 150 ⁇ g/mL) in an aqueous solvent.
- Solubility can be determined in any useful manner, such as an automated kinetic solubility method at any useful temperature (e.g., of from about 20° C. to 30° C., such as about 23° C.).
- a saturated pH-buffered aqueous solution is prepared, and the concentration of the compound is determined analytically (e.g., by gas chromatography, UV absorbance, liquid chromatography mass spectrometry (LC-MS), etc.), and the obtained spectrum is compared to a control spectrum for a precipitation-free reference solution.
- analytically e.g., by gas chromatography, UV absorbance, liquid chromatography mass spectrometry (LC-MS), etc.
- Exemplary aqueous solvents include phosphate buffered saline (PBS, 137 mM NaCl, 2.7 mM KCl, 10 mM sodium phosphate dibasic, 2 mM potassium phosphate monobasic and a pH of 7.4) or a cell medium (e.g., a cytopathic effect (CPE) medium including high glucose DMEM (Dulbecco's Modified Eagle's Medium) with 10% fetal bovine serum and 1 ⁇ penicillin streptomycin solution (Pen/Strep)).
- PBS phosphate buffered saline
- CPE cytopathic effect
- the compound has limited aqueous stability (e.g., of about 80% or less of a remaining amount of the compound after incubating in mouse plasma for about 3 hours; from about 20% to about 80% remaining amount of the compound after incubating in mouse plasma for about 3 hours; or from about 20% to 90% remaining amount of the compound after incubating at PBS, pH 7.4). Stability can be determined in any useful manner.
- the compound is dissolved in a solvent (e.g., at 10 ⁇ M in PBS at pH 7.4 with 1% dimethylsulfoxide (DMSO), human plasma, or mouse plasma) and analyzed at various timepoints (e.g., numerous timepoints from 0 to 48 hours).
- a solvent e.g., at 10 ⁇ M in PBS at pH 7.4 with 1% dimethylsulfoxide (DMSO), human plasma, or mouse plasma
- the concentration of the compound is determined analytically (e.g., by gas chromatography, UV absorbance, LC-MS, etc.), and absolute areas under the curve can be employed at each time point to determine the relative percent of the remaining parent compound.
- stability can be determined at about 37° C. with optional incubation at these temperature, centrifugation, and shaking.
- the compound has a computed hydrophobicity XLogP3-AA of from about 2 to about 10 (e.g., from 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 20, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, and 9 to 10).
- XLogP3-AA of from about 2 to about 10 (e.g., from 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 20, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to
- Hydrophobicity values can be determined in any useful manner, e.g., XLogP3-AA, which is a computationally generated octanol-water partition coefficient or distribution coefficient including an additive model, e.g., that can be determined according to Cheng T et al., “Computation of octanol-water partition coefficients by guiding an additive model with knowledge,” J. Chem. Inf. Model. 2007; 47:2140-8; ACD/LogP, which is another computationally generated octanol-water partition coefficient or distribution coefficient having correction factors, e.g., that can be determined according to Petrauskas A A et al., “ACD/Log P method description,” Perspect. Drug Discovery Des.
- XLogP3-AA which is a computationally generated octanol-water partition coefficient or distribution coefficient including an additive model, e.g., that can be determined according to Cheng T et al., “Computation of octanol
- TPSA topological polar surface area
- Non-limiting, exemplary antiviral compounds include, e.g., (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-nitro-N-phenylbenzamide; (E)-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-phenylbenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(4-methoxyphenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(2-fluorophenyl)-5-nitrobenzamide; (E)-4-chloro-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-phenylbenzamide; (E)-2-((1-ethyl-4-methylpiperazin-2-
- the compound or antiviral compound is from a class of quinazolinone-based inhibitors.
- the compound has a structure of formula (I), (II), or (III), or a salt thereof ( FIG. 2B ).
- R 2 includes any substituent including an aryl or heterocyclyl moiety.
- each R 2 is, independently, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, or optionally substituted alkheterocyclyl.
- each of R 1 , R 3 , R 4 , R 5 , R 7 , and R 8 is any useful substituent (e.g., any described herein, such as H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, optionally substituted alkheterocyclyl, halo, nitro, amino, azido, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, optionally substituted cycloalkyl, or optionally substituted spirocyclyl for two nearby R groups taken together).
- any useful substituent e.g., any described herein, such as H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, optionally substituted alkheterocyclyl, halo, nitro, amino, azid
- each R 1 is, independently, H or optionally substituted alkyl.
- each R 2 is, independently, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, or optionally substituted alkheterocyclyl.
- each R 4 , R 5 , R 7 , and R 8 is, independently, H, optionally substituted alkyl, halo, nitro, amino, azido, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl, or in which R 4 and R 5 , taken together, or R 7 and R 8 , taken together, form an optionally substituted spirocyclyl.
- each R 3 and R 6 is, independently, H, optionally substituted alkyl, halo, nitro, nitroso, amino, azido, carboxyl, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl.
- exemplary cargos include an acidic, basic, and hydrophobic drug (e.g., antiviral agents, antibiotic agents, etc.); a protein (e.g., antibodies, carbohydrates, etc.); a nucleic acid (e.g., DNA, RNA, small interfering RNA (siRNA), minicircle DNA (mcDNA), small hairpin RNA (shRNA), complementary DNA (cDNA), naked DNA, and plasmid, as well as chimeras, single-stranded forms, duplex forms, and multiplex forms thereof and including nucleic acid sequences encoding any of these and including one or more modified nucleic acids); a CRISPR component, a nuclease, a plasmid, a plasmid that encodes a CRISPR component, a ribonucleoprotein complex, a Cas enzyme or an ortholog or homolog thereof, a guide RNA, as well as a nucleic acid sequence encoding any of these or a complement thereof); a diagnostic
- the present invention relates, in part, to a particle having a core.
- the core can provide any useful benefit.
- the core provides a surface upon which a lipid layer can be supported.
- the core provides a charged surface that allows for electrostatic interactions with the cargo and/or the lipid layer, or a portion thereof.
- the core can be characterized in any useful manner.
- the core can be characterized by a first dimension (e.g., core circumference, pore size of the core, core diameter, core length, or core width).
- a core dimension e.g., core circumference, core diameter, core length, or core width, as well as an average or mean value for any of these
- exemplary values for a core dimension include, without limitation, greater than about 1 nm (e.g., greater than about 5 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 125 nm, 150 nm, 200 nm, 300 nm, 500 nm, 750 nm, 1 ⁇ m, 2 ⁇ m, 5 ⁇ m, 10 ⁇ m, 20 ⁇ m, or more), including of from about 5 nm to about 300 nm (e.g.
- the particle includes a porous core (e.g., a silica core that is spherical and ranges in diameter from about 10 nm to about 250 nm (e.g., having a mean diameter of about 150 nm)).
- a porous core e.g., a silica core that is spherical and ranges in diameter from about 10 nm to about 250 nm (e.g., having a mean diameter of about 150 nm)
- the silica core is monodisperse or polydisperse in size distribution.
- the core can be further characterized by an electrostatic property.
- the core has a negative charge (e.g., a net negative charge), such as a zeta potential of from about ⁇ 10 mV to about ⁇ 200 mV (e.g., from ⁇ 10 mV to ⁇ 100 mV, ⁇ 10 mV to ⁇ 75 mV, ⁇ 10 mV to ⁇ 50 mV, ⁇ 10 mV to ⁇ 30 mV, ⁇ 15 mV to ⁇ 100 mV, ⁇ 15 mV to ⁇ 75 mV, ⁇ 15 mV to ⁇ 50 mV, ⁇ 15 mV to ⁇ 30 mV, ⁇ 20 mV to ⁇ 100 mV, ⁇ 20 mV to ⁇ 75 mV, ⁇ 20 mV to ⁇ 50 mV, ⁇ 20 mV to ⁇ 30 mV, ⁇ 20 mV to ⁇ 100 mV, ⁇ 20 m
- the core can be porous.
- the pore has a dimension (e.g., average pore size, pore diameter, pore radius, pore circumference, pore length, pore width, or pore depth) that is greater than about 0.5 nm (e.g., of from about 0.5 nm to about 30 nm, including from 0.5 nm to 10 nm, 0.5 nm to 20 nm, 0.5 nm to 25 nm, 1 nm to 10 nm, 1 nm to 15 nm, 1 nm to 20 nm, 1 nm to 25 nm, 1 nm to 30 nm, 2 nm to 5 nm, 2 nm to 10 nm, 2 nm to 20 nm, 2 nm to 25 nm, or 2 nm to 30 nm).
- 0.5 nm e.g., average pore size, pore diameter, pore radius, pore circumference, pore length
- a particle or a portion thereof may have a variety of shapes and cross-sectional geometries that may depend, in part, upon the process used to produce the particles.
- the core or particle can be a nanoparticle (e.g., having a diameter less than about 1 ⁇ m) or a microparticle (e.g., having a diameter greater than or equal to about 1 ⁇ m).
- a core or particle may have a shape that is a sphere, a donut (toroidal), a rod, a tube, a flake, a fiber, a plate, a wire, a cube, or a whisker.
- a collection of cores may have two or more of the aforementioned shapes.
- a cross-sectional geometry of the core may be one or more of circular, ellipsoidal, triangular, rectangular, or polygonal.
- a core may consist essentially of non-spherical cores.
- such cores may have the form of ellipsoids, which may have all three principal axes of differing lengths, or may be oblate or prelate ellipsoids of revolution.
- Non-spherical cores alternatively may be laminar in form, wherein laminar refers to particles in which the maximum dimension along one axis is substantially less than the maximum dimension along each of the other two axes.
- Non-spherical cores may also have the shape of frusta of pyramids or cones, or of elongated rods.
- the cores may be irregular in shape.
- a plurality of cores may consist essentially of spherical cores.
- Particles and cores for use in the present invention may be PEGylated and/or aminated as otherwise described in Int. Pub. Nos. WO 2015/042268 and WO 2015/042279, which is incorporated herein by reference in their entirety.
- the particle size distribution (e.g., size of the core for the particle or a size of the silica carrier), according to the present invention, depends on the application, but is principally monodisperse (e.g., a uniform sized population varying no more than about 5-20% in diameter, as otherwise described herein).
- particles or cores can range, e.g., from around 1 nm to around 500 nm in size, including all integers and ranges there between.
- the size is measured as the longest axis of the core.
- the cores are from around 5 nm to around 500 nm and from around 10 nm to around 100 nm in size.
- the cores or particles are monodisperse and range in size from about 25 nm to about 300 nm.
- the sizes used preferably include 50 nm (+/ ⁇ 10 nm) and 150 nm (+/ ⁇ 15 nm), within a narrow monodisperse range, but may be more narrow in range.
- the pores can be from around 0.5 nm to about 25 nm in diameter, often about 1 to around 20 nm in diameter, including all integers and ranges there between. In one embodiment, the pores are from around 1 to around 10 nm in diameter. In one embodiment, around 90% of the pores are from around 1 to around 20 nm in diameter. In another embodiment, around 95% of the pores are around 1 to around 20 nm in diameter.
- preferred cores or particles according to the present invention are monodisperse and range in size from about 25 nm to about 300 nm; exhibit stability (colloidal stability); have single cell binding specification to the substantial exclusion of non-targeted cells; are anionic, neutral or cationic for specific targeting (preferably cationic); are optionally modified with agents such as PEI (polyethylene imine), NMe 3+ , dye, crosslinker, ligands (ligands provide neutral charge); and optionally, are used in combination with a cargo to be delivered to the target.
- PEI polyethylene imine
- NMe 3+ dye
- crosslinker ligands
- ligands provide neutral charge
- the present invention is directed to cores or particles of a particular size (diameter) ranging from about 0.5 to about 30 nm, about 1 nm to about 30 nm, often about 5 nm to about 25 nm (preferably, less than about 25 nm), often about 10 to about 20 nm, for administration in any useful route.
- these cores or particles are often monodisperse and provide colloidally stable compositions. These compositions can be used to target host cells because of enhanced biodistribution/bioavailability of these compositions, and optionally, specific cells, with a wide variety of therapeutic and/or diagnostic agents that exhibit varying release rates at the site of activity.
- the cores can be produced in any useful manner.
- cores are formed by templating with a surfactant, a cross-linked micelle, a detergent, or any other useful molecule (see, e.g., Gao F et al., J. Phys. Chem. B. 2009; 113:1796-804; Lin Y S et al., Chem. Mater. 2009; 21(17):3979-86; and Zhang K et al., J. Am. Chem. Soc. 2013 Feb. 20; 135(7):2427-30).
- cores are formed by dendritic growth (see, e.g., Shen D et al., Nano Lett. 2014 Feb.
- Each batch of cores or particles can be characterized in any useful manner, such as by assessment of size and surface charge using dynamic light scattering (DLS) (NIST-NCL PCC-1 and PCC-2) and electron microscopy (NIST-NCL PCC-7 and PCC-15) and verification of low endotoxin contamination per health industry product standards (NCL STE-1.1).
- Resultant cores can be further processed, such as by modifying core condensation (e.g., by using acidified ethanol for silica), modifying core surface charge (e.g., by use of amine-containing silanes, such as APTES), etc.
- the core can be formed of any useful material (e.g., a metal oxide, alum, silica, including mesoporous forms thereof).
- the core is composed of a mesoporous silica nanoparticle (MSN).
- MSN mesoporous silica nanoparticle
- the present invention relates to a lipid layer disposed around a core.
- the lipid layer can be characterized in any useful manner, such as by the thickness of the layer (e.g., of from about 5 nm to about 50 nm), the number of layers within the lipid layer (e.g., two, three, four, five, six, seven, or more lipid bilayers), and/or the net charge of the lipid layer (e.g., a net non-negative charge, such as a net positive charge; or as determined by the composition of the lipid layer, such as a layer formed by use of a liposome formulation having more than about 20 mol. % of a cationic lipid, such as any herein (e.g., DOTAP)).
- the thickness of the layer e.g., of from about 5 nm to about 50 nm
- the number of layers within the lipid layer e.g., two, three, four, five, six, seven, or more lipid bilayers
- the lipid layer can include any useful component, including a cationic lipid, a pegylated lipid, a zwitterionic lipid, and/or a cholesterol.
- the lipid layer can include any useful lipid or combination of lipids or component, such as one or more lipids selected from the group of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-[phosphor-L-serine] (DOPS), 1,2-dioleoyl-3-trimethylammonium-propane (18:1 DOTAP), 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DOPG), 1,2-
- sterols include cholesterol (e.g., from ovine wool or from plant sources), campestanol, campesterol, cholestanol, cholestenone, desmosterol, 7-dehydrodesmosterol, dehydroepiandrosterone (DHEA), desmosterol, diosgenin, FF-MAS (14-demethyl-14-dehydrolanosterol), lanosterol, lathosterol, pregnenolone, sitostanol, sitosterol, stigmasterol, zymosterol, zymostenol, zymosterone, as well as derivatives thereof, such as sulfates thereof, esters thereof, stereoisomers thereof, deuterated forms thereof, sulfonated forms thereof, phosphorylated forms thereof, unsaturated forms thereof, keto forms thereof, oxidized forms thereof, an oxysterol thereof, PEGylated forms thereof (e.g., cholesterol-(polyethylene glycol)
- Cores, lipids, and cargos can be PEGylated with a variety of polyethylene glycol-containing compositions as described herein.
- PEG molecules can have a variety of lengths and molecular weights and include, but are not limited to, PEG 200, PEG 1000, PEG 1500, PEG 2000, PEG 4600, PEG 5000, PEG 10,000, PEG-peptide conjugates or combinations thereof.
- the lipid layer includes DOPE and DOTAP.
- the lipid layer includes a zwitterionic lipid (e.g., DOPC, DPPC, DOPE, DPPE, DSPE, DLPC, DMPC, POPC, or SOPC) with an optional PEG (e.g., PEG, PEG-2000 PE, PEG conjugated to DOPE, PEG conjugated to DPPE, PEG conjugated to DSPE, etc.).
- a zwitterionic lipid e.g., DOPC, DPPC, DOPE, DPPE, DSPE, DLPC, DMPC, POPC, or SOPC
- PEG e.g., PEG, PEG-2000 PE, PEG conjugated to DOPE, PEG conjugated to DPPE, PEG conjugated to DSPE, etc.
- the lipid layer includes DOTAP and cholesterol in a 1:1 molar ratio.
- the lipid layer includes PEG.
- the lipid layer includes DOPE.
- the lipid layer includes DOTAP in combination with about 4 mol. % DOPE, about 47 mol. % cholesterol, and about 2 mol. % DSPE-PEth000.
- the lipid layer includes about 10 to about 50 mol. % DOTAP, about 40 to 50 mol. % cholesterol, about 0 to 40 mol. % DOPE, and about 1 to 5 mol. % of a PEGylated lipid.
- the lipid layer can be formed by employing any useful lipid formulation.
- a non-limiting exemplary formulation can include the following: about 1 mol. % to about 5 mol. % of a PEGylated lipid (e.g., from 1 mol. % to 3 mol. %, 1 mol. % to 4 mol. %, 2 mol. % to 3 mol. %, 2 mol. % to 4 mol. %, 2 mol. % to 5 mol. %, 3 mol. % to 4 mol. %, or 3 mol. % to 5 mol. %); about 30 mol. % to about 60 mol. % of a sterol (e.g., from 30 mol.
- % to 50 mol. % 35 mol. % to 50 mol. %, 35 mol. % to 60 mol. %, 40 mol. % to 50 mol. %, 40 mol. % to 60 mol. %, 45 mol. % to 50 mol. %, 45 mol. % to 60 mol. %, 50 mol. % to 60 mol. %, or 55 mol. % to 60 mol. %); about 20 mol. % to about 90 mol. % of a cationic lipid (e.g., from 20 mol. % to 30 mol. %, 20 mol. % to 40 mol. %, 20 mol. % to 50 mol.
- a cationic lipid e.g., from 20 mol. % to 30 mol. %, 20 mol. % to 40 mol. %, 20 mol. % to 50 mol.
- % to 90 mol. % 50 mol. % to 60 mol. %, 50 mol. % to 70 mol. %, 50 mol. % to 80 mol. %, 50 mol. % to 90 mol. %, 60 mol. % to 70 mol. %, 60 mol. % to 80 mol. %, 60 mol. % to 90 mol. %, 70 mol. % to 80 mol. %, 70 mol. % to 90 mol. %, or 80 mol. % to 90 mol. %); and about 0 mol. % to about 40 mol. % of a zwitterionic lipid (e.g., 0 mol.
- the ratio of the sterol to the cationic lipid is about 1:1.
- the lipid formulation includes about 2% of the PEGylated lipid.
- the lipid formulation includes about 30 mol. % to about 60 mol. % of the cationic lipid.
- the lipid formulation can include any useful lipid or component.
- exemplary PEGylated lipids include PEGylated DSPE (e.g., 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-X] (DSPE X) or N-[carbonyl-2′,3′-bis(methoxypolyethyleneglycol X)]-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-2arm PEGX)), PEGylated phosphoethanolamine (PE) (e.g., 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-X] (18:1 PEGX PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-X] (18:1 PEGX PE
- Exemplary sterols include, e.g., cholesterol, a derivative thereof, or any described herein.
- Exemplary zwitterionic lipids include DOPC, DPPC, DOPE, DPPE, POPC, DLPC, DSPC, DMPC, SOPC, or any described herein.
- Exemplary cationic lipids include 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-stearoyl-3-trimethylammonium-propane (18:0 TAP), 1,2-dipalmitoyl-3-trimethylammonium-propane (16:0 TAP), 1,2-dimyristoyl-3-trimethylammonium-propane (14:0 TAP), 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-amino-propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide (MVL5), ethylphosphocholine (ethyl PC) (e.g., 1,2-dimyristoleoyl-sn-glycer
- the lipid layer of the particle can be composed of lipids and components in an amount similar to that provided by the lipid formulation.
- an exemplary lipid formulation comprising about 47 mol. % of a cationic lipid can provide a lipid layer (for a lipid-coated particle) that comprises 47 mol. % of that cationic lipid.
- any composition provided for a lipid formulation herein also provides a composition for the lipid layer.
- the lipid-coated particle can include one or more cell targeting species, cell penetrating peptides, fusogenic peptides, and/or targeting peptides.
- Such species can be included within the cargo, configured to be expressed by a plasmid of the cargo, located within the lipid layer, and/or provided by an external surface of the lipid layer (e.g., provided by the outer lipid layer).
- the composition of the lipid layer can include one or more components that facilitate ligand orientation, maximize cellular interaction, provide lipid stability, and/or confer enhanced cellular entry.
- the targeting ligand can be a cell penetration peptide, a fusogenic peptide, or an endosomolytic peptide, which are peptides that aid a particle in translocating across a lipid bilayer, such as a cellular membrane or endosome lipid bilayer of the host cell.
- the targeting ligand is optionally crosslinked onto a lipid layer surface of the outer lipid layer.
- Endosomolytic peptides are a sub-species of fusogenic peptides as described herein.
- Representative and preferred electrostatic cell penetration (fusogenic) peptides include an 8 mer polyarginine (NH 2 -RRRRRRRR-COOH, SEQ ID NO:1), among others known in the art, which are included in or on particles in order to enhance the penetration of into cells.
- endosomolytic fusogenic peptides include HSWYG peptide (NH 2 -GLFHAIAHFIHGGWHGLIHGWYGGC-COOH, SEQ ID NO:2), RALA peptide (NH 2 -WEARLARALARALARHLARALARALRAGEA-COOH, SEQ ID NO:3), KALA peptide (NH 2 -WEAKLAKALAKALAKHLAKALAKALKAGEA-COOH), SEQ ID NO:4), GALA (NH 2 -WEAALAEALAEALAEHLAEALAEALEALAA-COOH, SEQ ID NO:5) and INF7 (NH 2 -GLFEAIEGFIENGWEGMIDGWYG-COOH, SEQ ID NO:6), or fragments thereof, among others.
- the targeting ligand includes an amino acid sequence having at least 80% sequence identity (e.g., at least 85%, 90%, 95%, or 99% sequence identity) to any one of SEQ ID NO:
- ligands can include a nuclear localization sequence (NLS), e.g., NH 2 -GNQSSNFGPMKGGNFGGRSSGPY GGGGQYFAKPRNQGGYGGC-COOH (SEQ ID NO:9), RRMKWKK (SEQ ID NO:10), PKKKRKV (SEQ ID NO:11), and KR[PAATKKAGQA]KKKK (SEQ ID NO:12), the NLS of nucleoplasmin, a prototypical bipartite signal comprising two clusters of basic amino acids, separated by a spacer of about 10 amino acids.
- NLS nuclear localization sequence
- Numerous other nuclear localization sequences are well known in the art.
- Preferred ligands which may be used to target cells include peptides, affibodies, and antibodies (including monoclonal and/or polyclonal antibodies).
- targeting ligands selected from the group consisting of Fc ⁇ from human IgG (which binds to Fc ⁇ receptors on macrophages and dendritic cells), human complement C3 (which binds to CR1 on macrophages and dendritic cells), ephrin B2 (which binds to EphB4 receptors on alveolar type II epithelial cells), SP94 peptide (which binds to unknown receptor(s) on hepatocyte-derived cells), and MET receptor binding peptide.
- Exemplary, non-limiting SP94 peptides include SP94 free peptide (H 2 N-SFSIILTPILPL-COOH, SEQ ID NO:13), a SP94 peptide modified with C-terminal Cys for conjugation (H 2 N-SFSIILTPILPLGGC-COOH, SEQ ID NO:14), and a further modified SP94 peptide (H 2 N-SFSIILTPILPLEEEGGC-COOH, SEQ ID NO:15).
- Exemplary MET binding peptides include ASVHFPP (SEQ ID NO:16), TATFWFQ (SEQ ID NO:17), TSPVALL (SEQ ID NO:18), IPLKVHP (SEQ ID NO:19), and WPRLTNM (SEQ ID NO:20).
- exemplary targeting ligands include poly-L-arginine, including (R) n , where 6 ⁇ n ⁇ 12, such as an R12 peptide (e.g., RRRRRRRRRRRR (SEQ ID NO:21)) or an R9 peptide (e.g., RRRRRRRRR (SEQ ID NO:22)); a poly-histidine-lysine, such as a (KH) 9 (e.g., KHKHKHKHKHKHKHKHKH (SEQ ID NO:23)); a Tat protein or derivatives and fragments thereof, such as RKKRRQRRR (SEQ ID NO:24), GRKKRRQRRRPQ (SEQ ID NO:25), GRKKRRQRRR (SEQ ID NO:26), GRKKRRQRRRPPQ (SEQ ID NO:27), YGRKKRRQRRR (SEQ ID NO:28), and RKKRRQRRRRKKRRQRRR (SEQ ID NO:29); a Cady protein or derivatives
- the targeting ligand includes an amino acid sequence having at least 80% sequence identity (e.g., at least 85%, 90%, 95%, or 99% sequence identity) to any one of SEQ ID NOs:10-12 and 21-49 or a fragment thereof (e.g., having a length of about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, or more amino acids).
- the lipid-coated particle can be characterized by any useful characteristic (e.g., overall charge, dimension, dispersity, etc.).
- one or more optional targeting ligands can be present in or on a lipid layer.
- the particle can have any useful dimension, such as diameter, circumference, length, width, height, etc.
- Exemplary values for dimensions include, without limitation, greater than about 10 nm (e.g., greater than about 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 125 nm, 150 nm, 200 nm, 300 nm, 500 nm, 750 nm, 1 ⁇ m, 2 ⁇ m, 5 ⁇ m, 10 ⁇ m, 20 ⁇ m, or more) or of from about 2 nm to 500 nm (e.g., from 2 nm to 50 nm, 2 nm to 100 nm, 2 nm to 150 nm, 2 nm to 200 nm, 2 nm to 300 nm, 2 nm to 400 nm, 10 nm to 50 nm, 10 nm to 100 nm, 10 nm to 150 nm, 10 nm to 200 nm, 10 n
- a plurality of particles is monodisperse, such as by having a polydispersity index (PdI) that is less than about 0.2 or by having a PdI that is of from about 0.05 to about 0.2 (e.g., from 0.05 to 0.1, 0.05 to 0.15, 0.1 to 0.15, 0.1 to 0.2, or 0.15 to 0.2).
- the monodisperse particles range in a size of from about 20 nm to about 300 nm (e.g., from 50 nm (+/ ⁇ 10 nm) to 150 nm (+/ ⁇ 15 nm)).
- the particle (or a plurality of particles) has a charge (or a net charge) that is near neutral (e.g., a zeta potential of from about +5 mV to ⁇ 5 mV).
- the present invention is directed to particles of a particular size (diameter) ranging from about 0.5 to about 30 nm, about 1 nm to about 30 nm, often about 5 nm to about 25 nm (preferably, less than about 25 nm), often about 10 to about 20 nm, for administration via intravenous, intramuscular, intraperitoneal, retro-orbital, and subcutaneous injection routes.
- These particles can be monodisperse and provide colloidally stable compositions.
- the lipid layer can have an appropriate charge (e.g., approximately net neutral charge), can include appropriate targeting ligands to promote their cell-specific binding and internalization, and can include useful ligand (e.g., to promote endosomal escape or nuclear localization within host cells).
- an appropriate charge e.g., approximately net neutral charge
- useful ligand e.g., to promote endosomal escape or nuclear localization within host cells.
- ligands include a peptide that binds to ephrin B2, which we identified using phage display, to target Vero cells; Fc ⁇ to target THP-1 cells and primary alveolar macrophages; the ‘GE11’ peptide (see, e.g., Li Z et al., FASEB J 2005; 19: 1978-85) to target A549 cells and primary alveolar epithelial cells; the ‘SP94’ peptide (see, e.g., Lo A et al., Molec. Cancer Therap.
- ligands include a peptide (e.g., a peptide zip code or a cell penetrating peptide), an endosomolytic peptide, an antibody (including fragments thereof), affibodies, a carbohydrate, an aptamer, a cluster of differentiation (CD) protein, or a self-associated molecular pattern (SAMP) (e.g., as described in Lambris J D et al., Nat. Rev. Microbiol. 2008; 6(2):132; and Poon I K H, Cell Death Differ. 2010; 17:381-97, each of which is incorporated herein by reference in its entirety).
- exemplary CD proteins include CD47 (OMIM Entry No.
- OMIM Entry No. 107271 a marker that prevents lysis by complement
- C1 inhibitor C1INH, OMIM Entry No. 606860, a marker that suppresses activation of the host's complement system
- CD200 OMIM Entry No. 155970, an immunosuppressive factor
- CD55 OMIM Entry No. 125240, a marker that inhibits the complement cascade
- CD46 OMIM Entry No. 120920, a marker that inhibits the complement cascade
- CD31 OMIM Entry No. 173445, an adhesion regulator and a negative regulator of platelet-collagen interactions.
- Any other useful ligand can be employed, such as those identified by the ‘BRASIL’ (Biopanning and Rapid Analysis of Selective Interactive Ligands) method (see, e.g., Giordano R J et al., Nat. Med. 2001; 7:1249-53; Giordano R J et al., Proc. Natl Acad. Sci. USA 2010; 107(11):5112-7; and Kolonin M G et al., Cancer Res. 2006; 66:34-40) to identify novel targeting peptides and single-chain variable fragments (scFvs) via phage display (see, e.g., Giordano R J et al., Chem. Biol.
- BRASIL Biopanning and Rapid Analysis of Selective Interactive Ligands
- the lipid-coated particles herein can be employed in any useful manner.
- the present particles can be adapted to recognize the target and, if needed, deliver the one or more cargos to treat that target.
- exemplary targets include a cell, a pathogen, an organ (e.g., dermis, vasculature, lymphoid tissue, liver, lung, spleen, kidneys, heart, brain, bone, muscle, etc.), a cellular target (e.g., targets of the subject, such as a human subject, including host tissue, host cytoplasm, host nucleus, etc., in any useful cell, such as e.g., hepatocytes, alveolar epithelial cells, and innate immune cells, etc.); as well as targets for exogenous cells and organisms, such as extracellular and/or intracellular components of a pathogen, e.g., bacteria), a molecular target (e.g., within the subject or the exogenous cell/organism, such as pathogen
- the particle is employed to target a host (e.g., a subject), a pathogen, or both (e.g., thereby treating the subject and/or the target).
- a pathogen include a bacterium, such as Bacillus (e.g., B. anthracis ), Enterobacteriaceae (e.g., Salmonella, Escherichia coli, Yersinia pestis, Klebsiella, and Shigella ), Yersinia (e.g., Y. pestis or Y. enterocolitica ), Staphylococcus (e.g., S.
- Bacillus e.g., B. anthracis
- Enterobacteriaceae e.g., Salmonella, Escherichia coli, Yersinia pestis, Klebsiella, and Shigella
- Yersinia e.g., Y. pestis or Y. enterocolitica
- Rickettsia e.g., R. rickettsii, R. prowazekii, or R. typhi
- Francisella tularensis Chlamydia psittaci, Coxiella burnetii , Mycoplasma (e.g., M. mycoides ), etc.
- mycotoxins, mold spores, or bacterial spores such as Clostridium botulinum and C.
- viruses including DNA or RNA viruses, such as Adenoviridae (e.g., adenovirus), Arenaviridae (e.g., Machupo virus), Bunyaviridae (e.g., Hantavirus or Rift Valley fever virus), Coronaviridae, Orthomyxoviridae (e.g., influenza viruses), Filoviridae (e.g., Ebola virus and Marburg virus), Flaviviridae (e.g., Japanese encephalitis virus, hepatitis C virus, and Yellow fever virus), Hepadnaviridae (e.g., hepatitis B virus), Herpesviridae (e.g., herpes simplex viruses, herpesvirus, cytomegalovirus, Epstein-Barr virus, or varicella zoster viruses), Papillomaviridae (e.g., papilloma viruses), Papovaviridae (e.g.
- Adenoviridae
- helminth such as cestodes (tapeworms), trematodes (flukes), or nematodes (roundworms, e.g., Ascaris lumbricoides, Trichuris trichiura, Necator americanus, or Ancylostoma duodenale ); a parasite (e.g., any protozoa or helminths described herein); or a fungus, such as Aspergilli, Candidae, Coccidioides immitis, and Cryptococci.
- Other pathogens include a multi-drug resistant (MDR) pathogen, such as MDR forms of any pathogen described herein.
- MDR multi-drug resistant
- the present invention can be employed to treat an infection (e.g., a viral infection).
- infections include an encephalitis infection, a viral infection, a bacterial infection, etc.
- Infections can arise from a virus, such as a mosquito-borne viral pathogen, an encephalitis virus (e.g., Venezuelan equine encephalitis virus (VEEV)), herpes virus (e.g., herpes simplex virus, varicella-zoster virus, and Epstein-Barr virus), rabies virus, poliovirus, measles virus, an arbovirus (e.g., St.
- a virus such as a mosquito-borne viral pathogen, an encephalitis virus (e.g., Venezuelan equine encephalitis virus (VEEV)), herpes virus (e.g., herpes simplex virus, varicella-zoster virus, and Epstein-Barr virus), rabies virus, poliovirus, measles virus, an
- Louis encephalitis virus and West Nile encephalitis virus bunyavirus (e.g., La Crosse strain, California encephalitis virus, etc.), arenavirus (e.g., lymphocytic choriomeningitis virus), reovirus (e.g., Colorado tick virus), henipavirus, flavivirus (e.g., Japanese encephalitis virus (JEV), St. Louis encephalitis virus, etc.), enterovirus, and Powassan virus.
- the infection can arise from bacteria, fungi, and/or protozoa.
- the present lipid-coated particles can be formulated in any useful manner.
- the formulation can be optimized for subcutaneous (SC), intranasal (IN), aerosol, intravenous (IV), intramuscular (IM), intraperitoneal (IP), oral, topical, transdermal, or retro-orbital delivery.
- Any useful dosages can be employed within the formulations.
- Exemplary dosages include, e.g., 200 mg/kg.
- the formulation or composition can include a plurality of particles (e.g., an effective amount thereof) and an optional pharmaceutically acceptable excipient (e.g., any described herein).
- the pharmaceutical composition includes a population of particles (e.g., any described herein) in an amount effective for modulating or modifying a target gene within a subject in combination with a pharmaceutically acceptable carrier, additive, or excipient.
- the composition further includes a drug, a therapeutic agent, etc., which is not disposed as cargo within the particle.
- the composition can be formulated in any useful manner with a plurality of particles.
- Such formulations can be included with any useful medium, excipient (e.g., lactose, saccharide, carbohydrate, mannitol, leucine, PEG, trehalose, etc.), additive, propellant, solution (e.g., aqueous solution, such as a buffer), additive, preservative, carrier (e.g., aqueous saline, aqueous dextrose, glycerol, or ethanol), binder (e.g., saccharide, cellulose preparation, starch paste, or methyl cellulose), filler, or disintegrator.
- excipient e.g., lactose, saccharide, carbohydrate, mannitol, leucine, PEG, trehalose, etc.
- propellant e.g., aqueous solution, such as a buffer
- solution e.g., aqueous solution, such
- compositions according to the present invention include an effective population of lipid-coated particles herein formulated to affect an intended result (e.g., immunogenic result, therapeutic result and/or diagnostic analysis, including the monitoring of therapy) formulated in combination with a pharmaceutically acceptable carrier, additive or excipient.
- the particles within the population of the composition may be the same or different depending upon the desired result to be obtained.
- Pharmaceutical compositions according to the present invention may also comprise an addition bioactive agent or drug, such as an antibiotic or antiviral agent.
- Formulations and compositions containing the particles according to the present invention may take the form of liquid, solid, semi-solid or lyophilized powder forms, such as, for example, solutions, suspensions, emulsions, sustained-release formulations, tablets, capsules, powders, suppositories, creams, ointments, lotions, aerosols, patches or the like, preferably in unit dosage forms suitable for simple administration of precise dosages.
- composition to be administered will contain a quantity of the selected compound in a pharmaceutically effective amount for therapeutic use in a biological system, including a patient or subject according to the present invention.
- VEEV Venezuelan Equine Encephalitis Virus
- VEEV In addition to its use as a bioterrorism agent, natural VEEV outbreaks result in equine and human infections in North and South America, causing high rates of fatality in equines (85%) and chronic neurological complications in humans [3-5]. Infected humans experience influenza-like symptoms, and 14% of infections result in neurological complications and sequelae, including disorientation, ataxia, depression, and convulsions [2, 5]. In one percent of cases, human infections result in mortality [4, 5]. Thus, developing strategies to inhibit VEEV infection is critical to minimizing fatalities and complications in cases of bioterrorism and natural outbreaks. For all these reasons, VEEV poses a major public health risk due to its amenability to use as a bioterrorism agent and its severe health consequences in humans and equines.
- ML336 this molecule was found to have a EC 90 of 170 nM against a VEEV vaccine strain (TC-83) and reduce viral titer by 630,000-fold at nanomolar concentrations.
- TC-83 VEEV vaccine strain
- intraperitoneal administration of ML336 to mice infected with TC-83 resulted in a 71% survival rate as compared to the 14% survival rate observed in untreated mice.
- ML336 has limited solubility (0.04 mg/mL in PBS, pH 7.4) and limited stability (reduction of 17% and 35% of drug in PBS and mouse plasma, respectively, after 3 hours) in aqueous solutions [4], potentially reducing its effectiveness.
- ML336 is a recently developed small molecule inhibitor of VEEV, shown to effectively reduce VEEV strain TC-83 both in vitro and in vivo, but its limited solubility and stability could hinder its use in future applications.
- a nanoparticle based platform to deliver ML336 for VEEV inhibition both in vitro and in vivo.
- LC-MSNs lipid-coated mesoporous silica nanoparticles
- MSNs Mesoporous silica nanoparticles
- MSNs have been used in drug delivery systems to improve drug stability and solubility, protect cargo, target specific tissues, and enhance drug circulation time and controlled release [12, 13].
- MSNs have a narrow size distribution and can be optimized for various drug delivery applications by tuning particle size, pore size, surface properties, and the porous structure [14, 15].
- Established methods enable formation of MSNs with uniform and tunable pore size, endowing MSNs with a large and uniform surface area for drug adsorption (600-1000 m 2 /g) [12-15].
- MSNs are stable in non-aqueous solutions and permit loading of hydrophobic drugs in organic solvents, giving them a distinct advantage over polymeric or liposomal nanoparticle delivery systems [18].
- MSNs are a promising carrier for ML336, drug-loaded MSNs can have low colloidal stability and are subject to aggregation in physiological solutions, reducing circulation time and preventing desirable cell uptake [19, 20]. In addition, premature release of cargo from MSNs can be problematic [21]. In order to overcome these challenges, we investigated the application of a lipid-based coating to the exterior of ML336-loaded MSNs.
- MSNs coated with supported lipid bilayers have been employed in drug and protein delivery applications to improve colloidal stability and subsequent circulation time, biocompatibility, cargo loading and release, and tissue-specific targeting [19, 21-23].
- lipid-coated MSNs lipid-coated MSNs (LC-MSNs)
- LC-MSNs lipid-coated MSNs
- the application of a supported lipid bilayer to the exterior of the MSN (essentially encapsulating the MSN with a liposome) can improve colloidal stability in physiological solutions [19, 20] and prevent cargo release prior to cell internalization or some other external trigger [21].
- a lipid bilayer coating offers an additional surface that can be functionalized independently of the MSN surface for tissue-specific targeting [19, 21, 22, 24].
- LC-MSNs harness the advantages and overcome the obstacles associated with MSNs and liposomes in one versatile platform for small molecule delivery.
- LC-MSN characterization revealed uniformly sized particles coated with a lipid bilayer and good colloidal stability, as assessed by dynamic light scatter analysis, zeta potential measurements, and cryogenic electron microscopy.
- the delivery vehicle was able to load and release ML336 in a manner that inhibited virus in vitro.
- LC-MSNs were found to load 20 ⁇ 3.4 ⁇ g ML336/mg LC-MSN and to release 6.6 ⁇ 1.3 ⁇ g/mg over the course of 24 hours.
- ML336-loaded LC-MSNs inhibited VEEV in vitro in a dose-dependent manner and by about 4-6 orders of magnitude as compared to untreated controls.
- in vitro studies suggested that additional release of ML336 occurs after cellular internalization, in which studies suggest that this was mediated through a clathrin-mediated endocytosis pathway.
- ML336 loaded LC-MSNs showed viral inhibition in an in vivo murine model of VEEV infection.
- In vivo safety studies in C3H/HeN mice shows that LC-MSNs were not toxic when dosed at 0.11 mg LC-MSNs/kg daily for four days.
- ML336-loaded LC-MSNs showed significant reduction in brain viral titer in VEEV TC-83 infected mice as compared to PBS treated controls.
- MSN fabrication and characterization Both small and large batch syntheses of monosized hexagonally-structured MSNs were prepared as previously described [19,41,42] with modifications. MSNs (up-scaled batch of hexagonal small pore particles) were synthesized in a large batch format by dissolving 1.45 g of cetyl trimethylammonium bromide (CTAB) (Sigma) in 750 mL of a 0.32 M aqueous ammonium hydroxide solution in a parafilm covered beaker (1 L). The beaker was placed in a 50° C. silicon oil bath for 2 hr (hours) with continuously stirring at high speed (650 rpm).
- CAB cetyl trimethylammonium bromide
- TEOS tetraethyl orthosilicate
- CTAB removal was achieved by resuspending particles in 100 mL of 6 g/L ammonium nitrate in ethanol. Tubes were placed in a 60° C. bath with sonication for 1.5 hr or at 40° C. with sonication for 30 min. Particles were collected by centrifugation, washed with ethanol (with 90% ethanol and then 100% ethanol; or with 95% ethanol), collected by centrifugation, resuspended in 100 mL of a 1% HCl in ethanol solution, and sonicated for 1.5 hr at 60° C. or twice for 30 min at 40° C.
- Particles were once again collected by centrifugation, washed with 90% ethanol and then by 100% ethanol, collected by centrifugation and resuspended in 40 mL of 100% ethanol.
- the MSN suspension was passed through a 1 ⁇ m filter to remove large aggregates and weighed after particle desiccation. Size and zeta potential were measured using a Zetasizer instrument (Malvern).
- Fluorescently labeled (Cy3) nanoparticles were synthesized in a small scale format by dissolving 250 mg of CTAB in 150 mL of 0.32 M ammonium hydroxide solution in a 250 ml beaker. The reaction was covered with parafilm and heated to 50° C. in a silicone oil bath for 1 hr with continuously stirring at high speed (650 rpm). A Cy3 dye solution was prepared by dissolving 3 mg of Cy3-NHS (ThermoFisher) in 1 ml of 100% ethanol using sonication, followed by the addition of 2.5 ⁇ L of APTES (Sigma). The Cy3 solution was allowed to incubate at room temperature without light for 1 hr.
- Cy3-NHS ThermoFisher
- CTAB removal was carried out by resuspending particles in 20 mL of a 6 g/L ammonium nitrite in ethanol, and sonicated at 40° C. for 20 min. Particles were collected by centrifugation, washed with 95% ethanol, collected by centrifugation, resuspended in 20 mL of an ethanolic HCl solution (1%) and sonicated twice for 20 min at 40° C. Cy3-labelled particles were washed with 90% ethanol followed by 100% ethanol, collected by centrifugation and resuspended in 12 mL of 100% ethanol.
- Non-Local Density Functional Theory was used to calculate pore size distributions and surface areas assuming the surface to be silica with cylindrical pores. Pore size distributions were also calculated according to the Barrett Joyner Halenda (BJH) method.
- BJH Barrett Joyner Halenda
- SE secondary electron
- ADF annular dark field
- ABSF annular bright field
- BF bright field
- LC-MSN fabrication and characterization Liposomes were prepared by combining 77.5 mol % 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 20 mol % cholesterol, and 2.5 mol % 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000) (Avanti Polar Lipids) in chloroform at a 5 mg/mL concentration.
- DSPC 1,2-distearoyl-sn-glycero-3-phosphocholine
- DSPE-PEG2000 Advanti Polar Lipids
- Lipid films were prepared using a rotary evaporator (Buchi Corp.), incubated under vacuum overnight, and rehydrated at a 5 mg/mL concentration in a 0.5 ⁇ PBS (Gibco/Life Technologies), 4 mM MgCl 2 solution for 30 minutes at 55° C.
- the lipid solution was purged with nitrogen for two minutes and then dispersed with an ultrasonication probe (Branson Sonifier, Emerson US). Lipids were sonicated under nitrogen at 10-12 watts for 4 min, followed by a 2 min rest period, repeated twice. Lipids were centrifuged at 16,000 RCF for 20 min to remove any debris deposited into the lipid solution from the sonication probe.
- Loaded LC-MSNs were prepared by resuspending 1 mg MSNs in 10 ⁇ L water followed by overnight incubation at 4° C. in 100 ⁇ l of 1 mg/mL ML336 (Caymen Chemicals) in dimethyl sulfoxide (DMSO) (100 ⁇ L DMSO for unloaded LC-MSN groups).
- DMSO dimethyl sulfoxide
- the resulting liposomes were combined with MSNs under bath sonication while pipetting at a 5:1 mass ratio of liposomes:nanoparticles. Particles were then centrifuged at 21,000 RCF to remove liposomes that were not fused with MSNs.
- LC-MSNs When used immediately, resulting LC-MSNs were rinsed twice by resuspending in 1 mL PBS, centrifuging at 21,000 RCF, and removing supernatant.
- particles were rinsed once in PBS and then resuspended in a 9 wt % sucrose solution in PBS, flash frozen in liquid nitrogen, and stored at ⁇ 80° C. Prior to use (e.g. cryo-EM analysis and all in vitro and in vivo studies), particles were thawed and rinsed once in PBS.
- LC-MSNs were prepared in 2 mg aliquots, combined, and redistributed into 1 or 1.5 mg aliquots before freezing for storage.
- DLS Dynamic light scattering
- PDI particle hydrodynamic diameter and polydispersity index
- zeta potential measurements were obtained using a Malvern Zetasizer.
- unloaded and loaded LC-MSNs were vitrified using Vitrobot (Thermo Fisher Scientific) as previously described [43]. Briefly, 3 ⁇ L of particles suspension was placed on a C-flat grid (Protochips, Inc.) with 2 ⁇ m diameter holes, blotted with filter paper, and plunged into liquid ethane for flash freezing. Frozen grids were stored under liquid nitrogen and were transferred to electron microscope JEM 2200FS (JEOL Ltd). Grids were imaged at 200 keV using DE-20 (Direct Detector Inc.) direct electron detector camera.
- the 2200FS microscope had Field Emission electron source and an omega-type electron energy filter to remove inelastically scattered electrons from image formation.
- the energy selecting slit was set to 20 eV.
- DE-20 camera was used in “movie” mode with frame rate of 25 frames/s.
- Off-line frame alignment was performed with DE_process_frames.py script provided by Direct Electron Inc. Images were collected at 40,000 ⁇ indicated magnification, the pixel size on the specimen scale corresponded to 1.5 ⁇ /pixel. Images were collected with 1.5 to 2.6 ⁇ m defocus range.
- Cryo-EM images were analyzed for lipid bilayer thickness using ImageJ. Fifty particles were analyzed from each of the ML336 loaded LC-MSN and unloaded LC-MSN groups.
- LC-MSN ML336 loading and release studies The concentration of ML336 was determined by correlating sample (supernatants) spectroscopic absorption measurements at 320 nm (Nanodrop, ThermoFisher Scientific) with ML336 standard curves prepared in the 5 mg/mL liposome solution (described above) or PBS ( FIG. 4C ).
- LC-MSN internalization studies All cells were maintained at 37° C. and 5% CO 2 . Cells were maintained in Dulbecco's Modified Eagle Medium (DMEM, Gibco/Life Technologies; HeLas) or Minimum Essential Medium a (aMEM, Gibco/Life Technologies; Veros) supplemented with 10 vol % FBS (Hyclone), 10,000 IU/mL penicillin, and 10,000 ⁇ g/ml streptomycin (MP Biomedicals).
- DMEM Dulbecco's Modified Eagle Medium
- aMEM Minimum Essential Medium a
- HeLa cells were plated overnight in complete medium on 12-well plates for image analysis or flow cytometry. Unless otherwise indicated, all inhibitors were purchased from EMD Millipore. The following inhibitors were initially resuspended in DMSO and were then diluted in complete medium to obtain the final concentrations indicated: the vacuolar H + -ATPase inhibitor bafilomycin A (BAF)(100 nM), the cationic amphiphilic compound chlorpromazine (CPZ; 6.5 ⁇ g/ml; Sigma), dynamin 2 inhibitor dynasore (DYN) (80 ⁇ M), wortmannin (Wort) (150 ng/ml), p21-activated kinase inhibitor III (IPA-3) (10 ⁇ M), and phorbol 12-myristate 13-acetate (PMA) (10 ⁇ M).
- BAF vacuolar H + -ATPase inhibitor bafilomycin A
- CPZ cationic amphiphilic compound chlorpromazine
- CPZ cati
- the cells were washed at 5 hr post-LC-MSN addition with PBS twice and then subjected to brightfield and fluorescent microscopy.
- HeLa cells were seeded overnight onto No. 1.5 glass cover slips in 6-well plates at a density of 0.75 ⁇ 10 5 cells per well. Fluorescent LC-MSNs were then added at 25 ⁇ g MSN/well for 45 min or 20 hr. After incubation, cells were washed with PBS, fixed with 4% paraformaldehyde in PBS for 15 min with prewarmed solutions followed by overnight storage at 4° C., washed twice with PBS, and made permeable with 0.1% Triton-X in PBS for 15 min.
- DMEM Dulbecco's Modified Eagle Medium
- aMEM Minimum Essential Medium a
- the TC-83 virus was obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH (NR-63), and was propagated in Vero cells.
- Cells were infected at a multiplicities of infection (MOI) of 0.1 and cultured for two days. Supernatant was collected and the concentration of plaque forming units (PFUs) was determined using a standard plaque assay with an agarose overlay (1:1 2 ⁇ Modified Eagle Medium (Gibco/Life Technologies; 8 vol % FBS, 10,000 IU/mL penicillin, and 10,000 m/ml streptomycin):1.5 wt % agarose (Invitrogen)). Cells were fixed and stained with an ethanol-based crystal violet solution (0.14 wt % crystal violet (Sigma-Aldrich), 21 vol % ethanol) and plaques were counted manually to determine PFU/mL.
- MOI multiplicities of infection
- PFUs plaque forming units
- HeLa cells at 80-90% confluency in 12 well plates were pretreated with 25 ⁇ g LC-MSNs in 100 ⁇ L Opti-MEM Reduced Serum Media (Gibco/Life Technologies) for 1 hr, then infected with TC-83 at 0.1 MOI for 30-60 min. Virus was then removed, cells were rinsed three times in PBS, and treatments were added back for the remainder of the time course in 1 mL DMEM. Supernatants were taken at 24, 48, and 72 hrs. Phase images were taken at the same timepoints using an inverted microscope (Olympus-IX70).
- LC-MSNs were incubated in Opti-MEM at a concentration of 2.5 ⁇ g/mL for 4 hrs at room temperature. LC-MSNs were then centrifuged at 20,000 RCF, and supernatant was collected. Particles were resuspended at 2.5 ⁇ g/mL in Opti-MEM and immediately added to cell cultures as described above. The supernatant of 25 ⁇ g of particles (100 ⁇ L) was also immediately added to cells.
- the concentration of PFUs in supernatants was determined using a standard plaque assay on VERO cells in 12 well plates as described above. Serial dilutions of supernatants were prepared in aMEM and used to infect cells for 30-60 min. Due to the minimum concentration of virions in supernatants required for detection in plaque assays, the limit of detection (indicated on each graph) was 100 PFU/mL.
- mice For free drug groups, 20 ⁇ L of 1 mg/mL ML336 in DMSO was mixed into 200 ⁇ L of 1 wt % carboxymethylcellulose sodium in PBS, resulting in 20 ⁇ g ML336 per injection, similar to what is loaded in 1 mg ML336-loaded LC-MSNs. Mice were injected twice a day IP for 4 days. Four hours after the first injection on day 1, the mice were infected with an intranasal instillation of 50 ⁇ l of TC-83 containing a total of 10 8 PFU.
- LC-MSN masses for both loaded and unloaded LC-MSN groups were increased to 1.5 mg.
- 15 ⁇ L of 2 mg/mL ML336 in DMSO was mixed into 185 ⁇ L of 1 wt % carboxymethylcellulose in PBS, resulting in 30 ⁇ g ML336 per injection, similar to what is loaded in 1.5 mg ML336-loaded LC-MSNs.
- Five mice from each group were euthanized and dissected for brain, spleen, kidney, liver, and serum. Organs were homogenized using disposable tissue grinders, and tissue lysate was assessed for viral load using a standard plaque assay as described above.
- ML336 was recently discovered to have antiviral drug properties against VEEV [4]. While proven effective both in vivo and in vitro, it has limited solubility in aqueous solution, necessitating a delivery vehicle to improve drug stability and enable controlled release.
- a hybrid liposome-mesoporous silica nanoparticle technology that takes advantage of the loading capabilities and uniformity of MSNs and the biocompatibility and retention capabilities of liposomes in one drug delivery platform ( FIG. 1C ) [19, 22, 26, 27].
- LC-MSNs see, e.g. [22]
- these particles have the potential to protect and control the release of ML336 as well as be modified for tissue-specific targeting in future iterations of the LC-MSN technology [19, 20].
- LC-MSNs formation was modified based on past methods [19, 20].
- monosized sub-150 nm MSNs were produced by up-scaling previous synthesis protocols.
- Optimized large batch synthesis procedures yielded highly homogeneous nanoparticles without alteration in structure or size.
- TEM transmission electron microscopy
- FIG. 3A the nanoparticles of approximately 75 nm were narrow in size distribution and displayed a hexagonal porous structure ( FIG. 3A , inset).
- FIG. 3B scanning electron microscopy (SEM) analysis was performed in order to highlight the 3D hexagonal shape of the MSNs and, importantly, the open porous structure.
- SEM scanning electron microscopy
- FIG. 3I surface accessible pores were clearly observed.
- N 2 sorption also provided evidence on the pore shape and its surface accessibility.
- the resulting isotherm ( FIG. 3I ) showed a steep increase in adsorption characteristic of a capillary condensation in mesopores capillary evaporation on the desorption branch, supporting the presence of uniform cylindrical mesopore open at both ends (surface-accessible).
- Liposomes were composed of 77.5:20:2.5 DSPC:Cholestrol:DSPE-PEG (2000) (mol %), a lipid composition chosen to ensure formation of a stable bilayer and to enhance colloidal stability of the resulting LC-MSNs.
- the primary lipid component, DSPC was chosen due to its saturated acyl chain, as previous work has indicated that unsaturated lipids may contribute to reduced colloidal stability of LCMSNs over time [19].
- Cholesterol was used to improve control over bilayer fluidity and leakage [22, 28], and a pegylated DSPE was included to increase circulation time and reduce protein adsorption to the LC-MSN surface [19, 22, 26, 27].
- a neutralized surface charge of particles is another measure of successful MSN-liposome fusion [31, 33], as a reduction of MSN surface charge is observed due to charge shielding of deprotonated silanols on the MSN surface by the zwitterionic lipid bilayer [19].
- uncoated MSNs had zeta potential of ⁇ 25.0 ⁇ 0.42 mV, similar to what has been reported in literature previously [13, 19, 20], which increased to nearly neutral levels when coated with a lipid bilayer ( ⁇ 0.263 ⁇ 0.41 mV, Table 1).
- MSNs are highly advantageous for small molecule delivery [12-15], aggregation of MSNs without surface modification or external coatings is commonly observed in high ionic strength physiologically relevant media due to a reduction in the Debye length and correspondingly the degree of electrostatic repulsion [12, 17, 20].
- ML336 loaded MSNs that were not coated with a lipid bilayer aggregated immediately in PBS (Table 1, FIG. 4A ).
- loaded LC-MSNs maintained colloidal stability for at least four days ( FIG. 4A ), indicating their utility in both in vitro and in vivo applications.
- zeta-potential, cryo-EM and stability studies indicate the formation of a complete, conformal and uniform lipid bilayer on ML336 loaded LC-MSNs.
- ML336 loading in LC-MSNs was determined to be about 20 ⁇ g ML336/mg LC-MSN, as measured by subtracting the amount of ML336 lost in the post-lipid-coating and loading washes from the total mass of ML336 loaded ( FIG. 4B-4D ).
- a linear burst release of ML336 was observed to occur in the first 4 hours, with little additional release thereafter ( FIG. 3E-3F ).
- LC-MSNs released about 6.6 ⁇ g ML336/mg LC-MSNs in 24 hours, which correlated to 34% release of ML336 loaded ( FIG. 3E-3F ; Table 2). No additional release was observed after 4 additional days.
- Cytotoxicity of ML336-loaded and unloaded LC-MSNs was assessed with HeLa cells. No visible differences in viability at 48 hrs was observed via LIVE/DEAD staining ( FIG. 6 ), in line with the high biocompatibility observed in cells treated with LC-MSNs previously [19, 33, 37] and with the limited toxicity observed when cells are treated with MSNs at a concentration less than 100 ⁇ g/mL [13].
- ML336-loaded LC-MSNs inhibited virus HeLa cells infected with the TC-83 virus were treated with ML336-loaded and unloaded LC-MSNs.
- ML336-loaded LC-MSNs Similar to soluble ML336, ML336-loaded LC-MSNs also inhibited virus in a dose-dependent manner, indicating that total ML336 release is proportional to LC-MSN mass and providing a method to tune drug dosage in a facile manner ( FIG. 7 ).
- ML336 loaded LC-MSNs significantly decreased viral load by at least 4 orders of magnitude after 24 hrs and 6 orders of magnitude after 48 and 72 hrs ( FIG. 8A-8B ), a greater reduction than previously observed for other small molecule VEEV inhibitors [9, 11] and similar to what has been observed for soluble ML336 [4]. Overall, these results indicate that ML336 loaded LCMSNs can successfully inhibit VEEV.
- TC-83 infected HeLa cells were then treated with LC-MSN supernatant and pre-released LC-MSNs and compared to cells treated with loaded LC-MSNs and untreated cells.
- LC-MSN supernatant inhibited virus at a similar level to ML336 loaded LC-MSNs until the 72 hr timepoint, at which point loaded LC-MSNs inhibited virus to a greater extent ( FIG. 8C , FIG. 9A-9D ). This indicates that while released ML336 remains bioactive, it is possible that LC-MSNs protect ML336 over time and/or continually release additional ML336 in a manner different from our test tube release studies.
- LC-MSNs While pre-released LC-MSNs inhibited virus in a similar manner to loaded LC-MSNs and LC-MSN supernatant at 24 hrs, by 48 hrs the extent of viral inhibition was significantly lower than cells treated with loaded LC-MSNs or LC-MSN supernatant. By 72 hrs, pre-released LC-MSNs showed no additional inhibition as compared to cells with no treatment ( FIG. 8C , FIG. 9A-9D ). This indicates that LC-MSNs release additional ML336 after the initial four hour burst release, which may either be undetectable in the loading and release studies or does not occur prior to cell internalization and disruption of the lipid bilayer [33].
- LC-MSNs conjugated with affinity ligands are known to enter cells using trafficking pathways of the targeting receptor.
- cholera toxin B conjugated LC-MSNs use caveolin-mediated endocytosis for internalization after binding the GM1 ganglioside receptor [20].
- LC-MSNs have also been formulated to avoid non-specific uptake in blood circulation [19]. However, LC-MSN uptake in static conditions represented in these studies is not well understood.
- LC-MSNs undergo cellular internalization through endocytosis
- fluorescent LC-MSNs containing Cy3-labeled MSN cores were used to facilitate visualization and quantitation of entry into HeLa cells while in the presence of various endocytosis inhibitors.
- Hela cells were treated with a pH dependent endocytosis inhibitor (bafilomycin, BAF), clathrin-mediated endocytosis inhibitors (dynamin II inhibitor dynasore, DYN, and chlorpromazine, CPZ), caveolae-medicated endocytosis inhibitors (phorbol 12-myristate 13-acetate, PMA and DYN), or macropinocytosis inhibitors (wortmannin, wort, and p21-activated kinase inhibitor III, IPA-3) for 1 hr prior to the addition of Cy3-labeled LC-MSNs. Cells were vigorously washed to remove free particles and then examined by microscopy methods.
- a pH dependent endocytosis inhibitor bafilomycin, BAF
- clathrin-mediated endocytosis inhibitors dynamin II inhibitor dynasore, DYN, and chlorpromazine, CPZ
- Cy3-labeled LC-MSNs were readily internalized by Hela cells under untreated or no inhibitor (NI) conditions. LC-MSNs were also clearly inhibited in the presence of BAF, CPZ, and DYN, but not by inhibitors of macropinocytosis (wort, IPA-3) nor caveolin-mediated endocytosis (PMA), suggesting the role of clathrin-mediated endocytosis in cellular internalization of LC-MSNs ( FIG. 10A ).
- LC-MSNs were internalized and not on the cell surface.
- high resolution confocal microscopy techniques were employed. HeLa cells were incubated with LC-MSNs for either 45 min or 20 hrs, washed, and then fixed for immunofluorescence staining. As shown in FIG. 10C-10D , HeLa cells incubated with LC-MSNs for 20 hrs were internalized as indicated by 3D rendering of LC-MSNs with actin, microtubules, and nuclei intracellular markers. An actin stain was used to mark the periphery of the cell as actin filaments are concentrated at the cell periphery and form a 3D network that determines cell shape.
- LC-MSNs were visualized as beneath actin filaments, on the same plane as the microtubules, and above the cell nucleus. Furthermore, these data indicated a time dependent mechanism of entry as particles were not seen intracellularly at 45 min ( FIG. 10C-10D ). Taken together, LC-MSNs enter cells through clathrin-mediated endocytosis and may provide a mechanism of additional drug/cargo release at the site of LC-MSN accumulation.
- LC-MSNs cell entry pathway as clathrin-mediated endocytosis may provide a mechanism to design additional drug/cargo release at the site of LC-MSN accumulation. Overall, these results motivated a further investigation of the ability of ML336 loaded LC-MSNs to inhibit virus in vivo.
- LC-MSNs As with all nanoparticle-based systems, the potential for LC-MSNs to dissolve, aggregate, and interact with living cells and animal tissues is dependent upon properties specific to their unique composition [14]. In addition, the toxicity of MSNs and LC-MSNs in general has yet to be fully assessed and can vary depending upon size and surface properties [12, 13, 18]. Thus, prior to conducting animal studies to evaluate antiviral efficacy, a safety study was conducted to determine if the LC-MSNs developed in this work affected mouse weight and survival over fifteen days. Mice were injected with 1 mg LC-MSNs twice daily for four days, and all animals survived treatment with no significant differences in total animal weight between LC-MSN and PBS treated groups ( FIG. 12A ).
- mice were treated with 1 mg loaded LC-MSNs, unloaded LC-MSNs, free ML336, or PBS only. Mice treated with ML336 loaded LC-MSNs showed greater survival than animals in the other three groups, though this result was not statistically significant ( FIG. 13A ). As we observed that LC-MSNs inhibited virus in a dose-dependent manner in in vitro studies ( FIG. 7 ), we were interested to see if an increased LC-MSN dose would improve animal outcomes. Thus, in the second set of animal studies, animals were dosed with 1.5 mg LC-MSNs. No differences were observed in spleen viral load ( FIG. 13C ), and very limited viral loads were detected in livers, kidneys, or serum ( FIG. 14A-14C ), similar to what has been observed in past studies characterizing TC-83 intranasal infection in C3H/HeN mice [8].
- LC-MSNs can prevent toxicity by 1) reducing the concentration required for drug efficacy, both through improvements in drug solubility/stability as well as circulation time [12, 19], and 2) protecting the cellular microenvironment from harmful cargo prior to triggered release, either through rupture of the lipid bilayer or a specific chemically triggered mechanism [22].
- the LC-MSN lipid bilayer can be modified to specifically target a tissue of interest, such as the blood-brain barrier [20, 22].
- LC-MSNs are particularly advantageous because properties of the lipid bilayer and the MSN can be independently tuned, enabling simultaneous optimization of the lipid bilayer for tissue-specific targeting and the core to maximize drug-specific loading.
- the studies presented here highlight the ability of drug loaded LC-MSNs to prevent viral infection in one particular case, but the versatility and modifiability of this technology will enable use of LC-MSNs to prevent viral infection in a variety of future applications.
- ML336 loaded LC-MSNs were successfully coated with a lipid bilayer, which significantly improved colloidal stability, and released cargo over the course of 4 hours. Viral loads were reduced by 4-6 orders of magnitude in TC-83 VEEV infected HeLa cells treated with ML336 loaded LC-MSNs, which was repeatable across several particle batches in different studies. Furthermore, in vitro studies could indicate the possibility of additional release of ML336 after cellular internalization via clathrin-mediated endocytosis and enhanced ML336 stability when loaded in LC-MSNs.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dispersion Chemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Optics & Photonics (AREA)
- Nanotechnology (AREA)
- Biomedical Technology (AREA)
- Physics & Mathematics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
- This application is a divisional application of, and discloses subject matter that is related to subject matters disclosed in, co-pending parent application U.S. patent application Ser. No. 16/443,316, filed Jun. 17, 2019, and entitled “LIPID-COATED PARTICLES FOR TREATING VIRAL INFECTIONS”, which claims priority to U.S. Provisional Patent Application No. 62/689,037, filed Jun. 22, 2018, and entitled “LIPID-COATED PARTICLES FOR TREATING VIRAL INFECTIONS”. The entireties of each of these applications are incorporated herein by reference.
- This invention was made with Government support under Contract No. DE-NA0003525 awarded by the United States Department of Energy/National Nuclear Security Administration. The Government has certain rights in the invention.
- A sequence listing appendix including an ASCII formatted file accompanies this application. The appendix includes a file named “SD14659.2 DIV_ST25.txt,” created on Apr. 14, 2021 (size of 12.7 kilobytes), which is hereby incorporated by reference in its entirety.
- The present invention relates to lipid-coated particles for treating viral infections, including viral encephalitis infections. In particular, an antiviral compound can be disposed within the lipid-coated particle, thereby providing an antiviral carrier. Methods of making and using such carriers are described herein.
- Promising therapeutics can often provide beneficial in vitro characteristics, yet fail during in vivo investigations. Various difficulties can arise, including adverse effects at therapeutic doses, limited bioavailability, and chemical instability. Accordingly, there is need for additional methodologies to improve delivery of such therapeutics to the desired site of action.
- The present invention relates to the use of a lipid-coated particle to improve the solubility and/or stability of a drug (e.g., an antiviral drug). As described herein, in one instance, lipid-coated mesoporous silica nanoparticles (LC-MSNs) were employed as delivery vehicles for antivirals with known solubility and stability issues. Without wishing to be limited by mechanism, the large surface area of the MSN core likely promotes loading of a hydrophobic or lipophilic drug, while the liposome coating could enable enhanced circulation time and biocompatibility, thereby providing an ideal carrier for antiviral ML336 drug delivery. Provides are determinations regarding colloidal stability; in vitro viral inhibition in a dose-dependent manner, as compared to untreated controls; and in vivo studies related to toxicity and efficacy in reducing brain viral titer of a virus (e.g., Venezuelan equine encephalitis virus (VEEV) TC-83 in mice). Overall, these results highlight the utility of LC-MSNs as drug delivery vehicles to treat viral infections.
- In a first aspect, the present invention features a method of increasing a stability and/or a solubility of a compound within an aqueous solution. In some embodiments, the method includes: incubating the compound with a core comprising a plurality of pores, thereby providing a loaded core; and coating the loaded core with a lipid layer, thereby provided a lipid-coated particle, wherein the stability and/or the solubility of the lipid-coated particle within the aqueous solution is greater than the stability and/or the solubility of the compound within the aqueous solution.
- In some embodiments, the compound has an aqueous solubility of from about 20 μg/mL to about 150 μg/mL in phosphate-buffered saline at a pH of 7.4 and/or a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- In a second aspect, the present invention features a method of treating a viral infection. In some embodiments, the method includes administering an effective amount of a lipid-coated particle to a subject. In other embodiments, the lipid-coated particle includes a porous core, a lipid layer, and an antiviral compound disposed within at least one pore of the porous core. In yet other embodiments, a concentration of the effective amount of the antiviral compound within the lipid-coated particle is less than a concentration of the effective amount of the antiviral compound alone.
- In some embodiments, the viral infection is an alphavirus infection. In other embodiments, the viral infection is an encephalitis infection.
- In a third aspect, the present invention features a method of treating viral encephalitis. In some embodiments, the method includes administering an effective amount of a lipid-coated particle to a subject. In some embodiments, the lipid-coated particle includes a porous core and an antiviral compound disposed within at least one pore of the porous core, where the effective amount of the lipid-coated particle provides a reduction of brain viral load, as compared to administration of the antiviral compound alone.
- In a fourth aspect, the present invention features a method of reducing brain viral load within a subject. In some embodiments, the method includes administering an effective amount of a lipid-coated particle to the subject. In particular embodiments, the lipid-coated particle comprises a porous core and an antiviral compound disposed within at least one pore of the porous core.
- In a fifth aspect, the present invention features an antiviral carrier including: a porous core comprising a plurality of pores; an antiviral compound disposed in at least one pore; and a lipid layer disposed around the porous core. In some embodiments, the antiviral compound has an aqueous solubility of from about 20 μg/mL to about 150 μg/mL in phosphate-buffered saline at a pH of 7.4 and/or a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- In some embodiments, the antiviral compound is present in an amount of from about 10 μg/mg to 50 μg/mg (μg of the compound per mg of the carrier).
- In some embodiments, the antiviral compound has a release rate of from about 3 μg/mg to about 20 μg/mg (μg of the compound per mg of the carrier) over a period of about 24 hours in vitro.
- In some embodiments, the lipid layer includes a zwitterionic lipid, a cholesterol or a derivative thereof, and a pegylated lipid.
- In some embodiments, the antiviral compound has an aqueous solubility of from about 20 μg/mL to about 150 μg/mL in phosphate-buffered saline at a pH of 7.4. In other embodiments, the antiviral compound has a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- In some embodiments, the antiviral compound has an EC50 value of from about 0.01 μM to about 1 μM as determined in a cellular assay.
- In some embodiments, the antiviral compound has an EC90 value of from about 100 nM to about 300 nM as determined in a cellular assay. In other embodiments, the antiviral compound is hydrophobic or lipophilic.
- In a sixth aspect, the present invention features a formulation including an antiviral (e.g., any described herein) and an optional pharmaceutically acceptable excipient.
- In any embodiment herein, the lipid-coated particle includes a porous core and an antiviral compound disposed within at least one pore of the porous core.
- In any embodiment herein, the lipid layer includes a zwitterionic lipid, a cholesterol or a derivative thereof, and a pegylated lipid.
- In any embodiment herein, the compound or the antiviral compound has an aqueous solubility of from about 20 μg/mL to about 150 μg/mL in phosphate-buffered saline at a pH of 7.4. In other embodiments, the compound or the antiviral compound has a stability of about 80% or less of a remaining amount of the compound after incubating in plasma for about 3 hours.
- In any embodiment herein, the compound or the antiviral compound has an EC50 value of from about 0.01 μM to about 1 μM as determined in a cellular assay. In other embodiments, the compound or the antiviral compound has an EC90 value of from about 100 nM to about 300 nM as determined in a cellular assay.
- In any embodiment herein, the compound or the antiviral compound is hydrophobic or lipophilic.
- In any embodiment herein, the compound or the antiviral compound has a structure of formula (I) or (II) or (III):
- or a salt thereof, wherein: each R2 is, independently, optionally substituted aryl (e.g., C4-18 aryl, including optionally substituted phenyl, such as a substituted p-phenyl), optionally substituted heterocyclyl, optionally substituted alkaryl (e.g., C1-6 alk-C4-18 aryl, including optionally substituted benzyl), or optionally substituted alkheterocyclyl; each R1, R3, R4, R5, R6, R7, and R8 is, independently, H, optionally substituted alkyl (e.g., C1-6 alkyl), halo, nitro, nitroso, amino, azido, carboxyl, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl, or in which R4 and R5, taken together, or R7 and R8, taken together, form an optionally substituted spirocyclyl.
- In some embodiments, each R4, R5, R7, and R8 is, independently, H, optionally substituted alkyl, halo, nitro, amino, azido, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl, or in which R4 and R5, taken together, or R7 and R8, taken together, form an optionally substituted spirocyclyl; and each R3 and R6 is, independently, H, optionally substituted alkyl, halo, nitro, nitroso, amino, azido, carboxyl, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl.
- Definitions
- As used herein, the term “about” means +/−10% of any recited value. As used herein, this term modifies any recited value, range of values, or endpoints of one or more ranges.
- By “alkaryl” is meant an aryl group, as defined herein, attached to the parent molecular group through an alkylene group, as defined herein. Similarly, by the term “alkheteroaryl” is meant a heteroaryl group, as defined herein, attached to the parent molecular group through an alkylene group. Other groups preceded by the prefix “alk-” are defined in the same manner. The alkaryl group can be substituted or unsubstituted. For example, the alkaryl group can be substituted with one or more substitution groups, as described herein for alkyl and/or aryl. Exemplary unsubstituted alkaryl groups are of from 7 to 16 carbons (C7-16 alkaryl), as well as those having an alkylene group with 1 to 6 carbons and an aryl group with 4 to 18 carbons (i.e., C1-6 alk-C4-18 aryl).
- By “alkheterocyclyl” represents a heterocyclyl group, as defined herein, attached to the parent molecular group through an alkylene group, as defined herein. Exemplary unsubstituted alkheterocyclyl groups are of from 2 to 14 carbons.
- By “alkyl” and the prefix “alk” is meant a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. The alkyl group can be cyclic (e.g., C3-24 cycloalkyl) or acyclic. The alkyl group can be branched or unbranched. The alkyl group can also be substituted or unsubstituted. For example, the alkyl group can be substituted with one, two, three or, in the case of alkyl groups of two carbons or more, four substituents independently selected from the group consisting of: (1) C1-6 alkoxy (e.g., —OAk, in which Ak is an alkyl group, as defined herein); (2) C1-6 alkylsulfinyl (e.g., —S(O)Ak, in which Ak is an alkyl group, as defined herein); (3) C1-6 alkylsulfonyl (e.g., —SO2Ak, in which Ak is an alkyl group, as defined herein); (4) amino (e.g., —NRN1RN2, where each of RN1 and RN2 is, independently, H or optionally substituted alkyl, or RN1 and RN2, taken together with the nitrogen atom to which each are attached, form a heterocyclyl group); (5) aryl; (6) arylalkoxy (e.g., —OALAr, in which AL is an alkylene group and Ar is an aryl group, as defined herein); (7) aryloyl (e.g., —C(O)Ar, in which Ar is an aryl group, as defined herein); (8) azido (e.g., an —N3 group); (9) cyano (e.g., a —CN group); (10) carboxyaldehyde (e.g., a —C(O)H group); (11) C3-8 cycloalkyl; (12) halo; (13) heterocyclyl (e.g., a 5-, 6- or 7-membered ring, unless otherwise specified, containing one, two, three, or four non-carbon heteroatoms (e.g., independently selected from the group consisting of nitrogen, oxygen, phosphorous, sulfur, or halo)); (14) heterocyclyloxy (e.g., —OHet, in which Het is a heterocyclyl group); (15) heterocyclyloyl (e.g., —C(O)Het, in which Het is a heterocyclyl group); (16) hydroxyl (e.g., a —OH group); (17) N-protected amino; (18) nitro (e.g., an —NO2 group); (19) oxo (e.g., an ═O group); (20) C3-8 spirocyclyl (e.g., an alkylene diradical, both ends of which are bonded to the same carbon atom of the parent group to form a spirocyclyl group); (21) C1-6 thioalkoxy (e.g., —SAk, in which Ak is an alkyl group, as defined herein); (22) thiol (e.g., an —SH group); (23) —CO2RA, where RA is selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) C4-18 aryl, and (d) C1-6 alk-C4-18 aryl; (24) —C(O)NBRC, where each of RB and RC is, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) C4-18 aryl, and (d) C1-6 alk-C4-18 aryl; (25) —SO2O, where RD is selected from the group consisting of (a) C1-6 alkyl, (b) C4-18 aryl, and (c) C1-6 alk-C4-18 aryl; (26) —SO2NRERF, where each of RE and RF is, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) C4-18 aryl, and (d) C1-6 alk-C4-18 aryl; and (27) —NRGRH, where each of RG and RH is, independently, selected from the group consisting of (a) hydrogen, (b) an N-protecting group, (c) C1-6 alkyl, (d) C2-6 alkenyl, (e) C2-6 alkynyl, (f) C4-18 aryl, (g) C1-6 alk-C4-18 aryl, (h) C3-8 cycloalkyl, and (i) C1-6 alk-C3-8 cycloalkyl, wherein in one embodiment no two groups are bound to the nitrogen atom through a carbonyl group or a sulfonyl group. The alkyl group can be a primary, secondary, or tertiary alkyl group substituted with one or more substituents (e.g., one or more halo or alkoxy). In some embodiments, the unsubstituted alkyl group is a C1-3, C1-6, C1-12, C1-16, C1-18, C1-20, or C1-24 alkyl group.
- By “alkylene” is meant a multivalent (e.g., bivalent, trivalent, tetravalent, etc.) form of an alkyl group, as described herein. Exemplary alkylene groups include methylene, ethylene, propylene, butylene, etc. In some embodiments, the alkylene group is a C1-3, C1-6, C1-12, C1-16, C1-18, C1-20, C1-24, C2-3, C2-6, C2-12, C2-16, C2-18, C2-20, or C2-24 alkylene group. The alkylene group can be branched or unbranched. The alkylene group can also be substituted or unsubstituted. For example, the alkylene group can be substituted with one or more substitution groups, as described herein for alkyl.
- By “amino” is meant —NRN1RN2, where each of RN1 and RN2 is, independently, H or optionally substituted alkyl, or RN1 and RN2, taken together with the nitrogen atom to which each are attached, form a heterocyclyl group, as defined herein.
- By “aryl” is meant a group that contains any carbon-based aromatic group including, but not limited to, benzyl, naphthalene, phenyl, biphenyl, phenoxybenzene, and the like. The term “aryl” also includes “heteroaryl,” which is defined as a group that contains an aromatic group that has at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus. Likewise, the term “non-heteroaryl,” which is also included in the term “aryl,” defines a group that contains an aromatic group that does not contain a heteroatom. The aryl group can be substituted or unsubstituted. The aryl group can be substituted with one, two, three, four, or five substituents independently selected from the group consisting of: (1) C1-6 alkanoyl (e.g., —C(O)Ak, in which Ak is an alkyl group, as defined herein); (2) C1-6 alkyl; (3) C1-6 alkoxy (e.g., —OAk, in which Ak is an alkyl group, as defined herein); (4) C1-6 alkoxy-C1-6 alkyl (e.g., an alkyl group, which is substituted with an alkoxy group —OAk, in which Ak is an alkyl group, as defined herein); (5) C1-6 alkylsulfinyl (e.g., —S(O)Ak, in which Ak is an alkyl group, as defined herein); (6) C1-6 alkylsulfinyl-C1-6 alkyl (e.g., an alkyl group, which is substituted by an alkylsulfinyl group —S(O)Ak, in which Ak is an alkyl group, as defined herein); (7) C1-6 alkylsulfonyl (e.g., —SO2Ak, in which Ak is an alkyl group, as defined herein); (8) C1-6 alkylsulfonyl-C1-6 alkyl (e.g., an alkyl group, which is substituted by an alkylsulfonyl group —SO2Ak, in which Ak is an alkyl group, as defined herein); (9) aryl; (10) amino (e.g., —NRN1RN2, where each of RN1 and RN2 is, independently, H or optionally substituted alkyl, or RN1 and RN2, taken together with the nitrogen atom to which each are attached, form a heterocyclyl group); (11) C1-6 aminoalkyl (e.g., meant an alkyl group, as defined herein, substituted by an amino group); (12) heteroaryl; (13) C1-6 alk-C4-18 aryl (e.g., —ALAr, in which AL is an alkylene group and Ar is an aryl group, as defined herein); (14) aryloyl (e.g., —C(O)Ar, in which Ar is an aryl group, as defined herein); (15) azido (e.g., an —N3 group); (16) cyano (e.g., a —CN group); (17) C1-6 azidoalkyl (e.g., a —N3 azido group attached to the parent molecular group through an alkyl group, as defined herein); (18) carboxyaldehyde (e.g., a —C(O)H group); (19) carboxyaldehyde-C1-6 alkyl (e.g., —ALC(O)H, in which AL is an alkylene group, as defined herein); (20) C3-8 cycloalkyl; (21) C1-6 alk-C3-8 cycloalkyl (e.g., —ALCy, in which AL is an alkylene group and Cy is a cycloalkyl group, as defined herein); (22) halo (e.g., F, Cl, Br, or I); (23) C1-6 haloalkyl (e.g., an alkyl group, as defined herein, substituted with one or more halo); (24) heterocyclyl; (25) heterocyclyloxy (e.g., —OHet, in which Het is a heterocyclyl group); (26) heterocyclyloyl (e.g., —C(O)Het, in which Het is a heterocyclyl group); (16) hydroxyl (e.g., a —OH group); (27) hydroxyl (e.g., a —OH group); (28) C1-6 hydroxyalkyl (e.g., an alkyl group, as defined herein, substituted by one to three hydroxyl groups, with the proviso that no more than one hydroxyl group may be attached to a single carbon atom of the alkyl group); (29) nitro (e.g., an —NO2 group); (30) C1-6 nitroalkyl (e.g., an alkyl group, as defined herein, substituted by one to three nitro groups); (31) N-protected amino; (32) N-protected amino-C1-6 alkyl; (33) oxo (e.g., an ═O group); (34) C1-6thioalkoxy (e.g., —SAk, in which Ak is an alkyl group, as defined herein); (35) thio-C1-6 alkoxy-C1-6 alkyl (e.g., an alkyl group, which is substituted by an thioalkoxy group —SAk, in which Ak is an alkyl group, as defined herein); (36) —(CH2)rCO2RA, where r is an integer of from zero to four, and RA is selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) C4-18 aryl, and (d) C1-6 alk-C4-18 aryl; (37) —(CH2)rCONRHRC, where r is an integer of from zero to four and where each RB and RC is independently selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) C4-18 aryl, and (d) C1-6 alk-C4-18 aryl; (38) —(CH2)rSO2RD, where r is an integer of from zero to four and where RH is selected from the group consisting of (a) C1-6 alkyl, (b) C4-18 aryl, and (c) C1-6 alk-C4-18 aryl; (39) —(CH2)rSO2NRERF, where r is an integer of from zero to four and where each of RE and RF is, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) C4-18 aryl, and (d) C1-6 alk-C4-18 aryl; (40) —(CH2)rNRGRH, where r is an integer of from zero to four and where each of RG and RH is, independently, selected from the group consisting of (a) hydrogen, (b) an N-protecting group, (c) C1-6 alkyl, (d) C2-6 alkenyl, (e) C2-6 alkynyl, (f) C4-18 aryl, (g) C1-6 alk-C4-18 aryl, (h) C3-8 cycloalkyl, and (i) C1-6 alk-C3-8 cycloalkyl, wherein in one embodiment no two groups are bound to the nitrogen atom through a carbonyl group or a sulfonyl group; (41) thiol; (42) perfluoroalkyl (e.g., an alkyl group, as defined herein, having each hydrogen atom substituted with a fluorine atom); (43) perfluoroalkoxy (e.g., —ORf, in which Rf is an alkyl group, as defined herein, having each hydrogen atom substituted with a fluorine atom); (44) aryloxy (e.g., —OAr, where Ar is an optionally substituted aryl group, as described herein); (45) cycloalkoxy (e.g., —OCy, in which Cy is a cycloalkyl group, as defined herein); (46) cycloalkylalkoxy (e.g., —OALCy, in which AL is an alkylene group and Cy is a cycloalkyl group, as defined herein); and (47) arylalkoxy (e.g., —OALAr, in which AL is an alkylene group and Ar is an aryl group, as defined herein). In particular embodiments, an unsubstituted aryl group is a C4-18, C4-14, C4-12, C4-10, C6-18, C6-14, C6-12, or C6-10 aryl group.
- By “azido” is meant an —N3 group.
- By “carboxyl” is meant a —CO2H group.
- By “cyano” is meant a —CN group.
- By “cycloalkyl” is meant a monovalent saturated or unsaturated non-aromatic cyclic hydrocarbon group of from three to eight carbons, unless otherwise specified, and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[2.2.1.]heptyl and the like. The cycloalkyl group can also be substituted or unsubstituted. For example, the cycloalkyl group can be substituted with one or more groups including those described herein for alkyl.
- By “halo” is meant F, Cl, Br, or I.
- By “haloalkyl” is meant an alkyl group, as defined herein, substituted with one or more halo.
- By “heteroalkyl” is meant an alkyl group, as defined herein, containing one, two, three, or four non-carbon heteroatoms (e.g., independently selected from the group consisting of nitrogen, oxygen, phosphorous, sulfur, or halo).
- By “heteroaryl” is meant a subset of heterocyclyl groups, as defined herein, which are aromatic, i.e., they contain 4n+2 pi electrons within the mono- or multicyclic ring system.
- By “heterocyclyl” is meant a 5-, 6- or 7-membered ring, unless otherwise specified, containing one, two, three, or four non-carbon heteroatoms (e.g., independently selected from the group consisting of nitrogen, oxygen, phosphorous, sulfur, or halo). The 5-membered ring has zero to two double bonds and the 6- and 7-membered rings have zero to three double bonds. The term “heterocyclyl” also includes bicyclic, tricyclic and tetracyclic groups in which any of the above heterocyclic rings is fused to one, two, or three rings independently selected from the group consisting of an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, and another monocyclic heterocyclic ring, such as indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, benzofuryl, benzothienyl and the like. Heterocyclics include thiiranyl, thietanyl, tetrahydrothienyl, thianyl, thiepanyl, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, pyrrolyl, pyrrolinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, homopiperidinyl, pyrazinyl, piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidiniyl, morpholinyl, thiomorpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, furyl, thienyl, thiazolidinyl, isothiazolyl, isoindazoyl, triazolyl, tetrazolyl, oxadiazolyl, uricyl, thiadiazolyl, pyrimidyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, dihydroindolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, pyranyl, dihydropyranyl, dithiazolyl, benzofuranyl, benzothienyl, and the like.
- By “hydroxyl” is meant —OH.
- By “hydroxyalkyl” is meant an alkyl group, as defined herein, substituted by one to three hydroxyl groups, with the proviso that no more than one hydroxyl group may be attached to a single carbon atom of the alkyl group and is exemplified by hydroxymethyl, dihydroxypropyl, and the like.
- By “nitro” is meant an —NO2 group.
- By “nitroalkyl” is meant an alkyl group, as defined herein, substituted by one to three nitro groups.
- By “nitroso” is meant an —NO group.
- By “perfluoroalkyl” is meant an alkyl group, as defined herein, having each hydrogen atom substituted with a fluorine atom. Exemplary perfluoroalkyl groups include trifluoromethyl, pentafluoroethyl, etc.
- By “protecting group” is meant any group intended to protect a reactive group against undesirable synthetic reactions. Commonly used protecting groups are disclosed in “Greene's Protective Groups in Organic Synthesis,” John Wiley & Sons, New York, 2007 (4th ed., eds. P. G. M. Wuts and T. W. Greene), which is incorporated herein by reference. O-protecting groups include an optionally substituted alkyl group (e.g., forming an ether with reactive group O), such as methyl, methoxymethyl, methylthiomethyl, benzoyloxymethyl, t-butoxymethyl, etc.; an optionally substituted alkanoyl group (e.g., forming an ester with the reactive group O), such as formyl, acetyl, chloroacetyl, fluoroacetyl (e.g., perfluoroacetyl), methoxyacetyl, pivaloyl, t-butylacetyl, phenoxyacetyl, etc.; an optionally substituted aryloyl group (e.g., forming an ester with the reactive group O), such as —C(O)—Ar, including benzoyl; an optionally substituted alkylsulfonyl group (e.g., forming an alkylsulfonate with reactive group O), such as —SO2-RS1, where RS1 is optionally substituted C1-12 alkyl, such as mesyl or benzylsulfonyl; an optionally substituted arylsulfonyl group (e.g., forming an arylsulfonate with reactive group O), such as —SO2-RS4, where RS4 is optionally substituted C4-18 aryl, such as tosyl or phenylsulfonyl; an optionally substituted alkoxycarbonyl or aryloxycarbonyl group (e.g., forming a carbonate with reactive group O), such as —C(O)—ORT1, where RT1 is optionally substituted C1-12 alkyl or optionally substituted C4-18 aryl, such as methoxycarbonyl, methoxymethylcarbonyl, t-butyloxycarbonyl (Boc), or benzyloxycarbonyl (Cbz); or an optionally substituted silyl group (e.g., forming a silyl ether with reactive group O), such as —Si—(RT2)3, where each RT2 is, independently, optionally substituted C1-12 alkyl or optionally substituted C4-18 aryl, such as trimethylsilyl, t-butyldimethylsilyl, or t-butyldiphenylsilyl. N-protecting groups include, e.g., formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, Boc, and Cbz. Such protecting groups can employ any useful agent to cleave the protecting group, thereby restoring the reactivity of the unprotected reactive group.
- By “salt” is meant an ionic form of a compound or structure (e.g., any formulas, compounds, or compositions described herein), which includes a cation or anion compound to form an electrically neutral compound or structure. Salts (e.g., simple salts having binary compounds, double salts, triple salts, etc.) are well known in the art. For example, salts are described in Berge S M et al., “Pharmaceutical salts,” J. Pharm. Sci. 1977 January; 66(1):1-19; International Union of Pure and Applied Chemistry, “Nomenclature of Inorganic Chemistry,” Butterworth & Co. (Publishers) Ltd., London, England, 1971 (2nd ed.); and in “Handbook of Pharmaceutical Salts: Properties, Selection, and Use,” Wiley-VCH, April 2011 (2nd rev. ed., eds. P. H. Stahl and C. G. Wermuth). The salts can be prepared in situ during the final isolation and purification of the compounds of the invention or separately by reacting the free base group with a suitable organic acid (thereby producing an anionic salt) or by reacting the acid group with a suitable metal or organic salt (thereby producing a cationic salt). Representative anionic salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, camphorate, camphorsulfonate, chloride, citrate, cyclopentanepropionate, digluconate, dihydrochloride, diphosphate, dodecylsulfate, edetate, ethanesulfonate, fumarate, glucoheptonate, glucomate, glutamate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, hydroxyethanesulfonate, hydroxynaphthoate, iodide, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methylbromide, methylnitrate, methyl sulfate, mucate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, theophyllinate, thiocyanate, triethiodide, toluenesulfonate, undecanoate, valerate salts, and the like. Representative cationic salts include metal salts, such as alkali or alkaline earth salts, e.g., barium, calcium (e.g., calcium edetate), lithium, magnesium, potassium, sodium, and the like; other metal salts, such as aluminum, bismuth, iron, and zinc; as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, pyridinium, and the like. Other cationic salts include organic salts, such as chloroprocaine, choline, dibenzylethylenediamine, diethanolamine, ethylenediamine, methylglucamine, and procaine.
- By “spirocyclyl” is meant an alkylene diradical, both ends of which are bonded to the same carbon atom of the parent group to form a spirocyclyl group and also a heteroalkylene diradical, both ends of which are bonded to the same atom.
- By “micro” is meant having at least one dimension that is less than 1 mm. For instance, a microstructure (e.g., any structure described herein, such as a microparticle) can have a length, width, height, cross-sectional dimension, circumference, radius (e.g., external or internal radius), or diameter that is less than 1 mm.
- By “nano” is meant having at least one dimension that is less than 1 μm. For instance, a nanostructure (e.g., any structure described herein, such as a nanoparticle) can have a length, width, height, cross-sectional dimension, circumference, radius (e.g., external or internal radius), or diameter that is less than 1 μm.
- The phrase “effective average particle size” as used herein to describe a multiparticulate (e.g., a porous nanoparticulate) means that at least 50% of the particles therein are of a specified size. Accordingly, “effective average particle size of less than about 2,000 nm in diameter” means that at least 50% of the particles therein are less than about 2,000 nm in diameter. In certain embodiments, nanoparticulates have an effective average particle size of less than about 2,000 nm (i.e., 2 microns), less than about 1,900 nm, less than about 1,800 nm, less than about 1,700 nm, less than about 1,600 nm, less than about 1,500 nm, less than about 1,400 nm, less than about 1,300 nm, less than about 1,200 nm, less than about 1,100 nm, less than about 1,000 nm, less than about 900 nm, less than about 800 nm, less than about 700 nm, less than about 600 nm, less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 250 nm, less than about 200 nm, less than about 150 nm, less than about 100 nm, less than about 75 nm, or less than about 50 nm, as measured by light-scattering methods, microscopy, or other appropriate methods. In certain aspects of the present invention, the particles are monodisperse and generally no greater than about 50 nm in average diameter, often less than about 30 nm in average diameter, as otherwise described herein. The term “D50” refers to the particle size below which 50% of the particles in a multiparticulate fall. Similarly, the term “D90” refers to the particle size below which 90% of the particles in a multiparticulate fall.
- The term “monodisperse” is used as a standard definition established by the National Institute of Standards and Technology (NIST) (Particle Size Characterization, Special Publication 960-1, January 2001) to describe a distribution of particle size within a population of particles, in this case nanoparticles, which particle distribution may be considered monodisperse if at least 90% of the distribution lies within 5% of the median size. See, e.g., Takeuchi S et al., Adv. Mater. 2005; 17(8): 1067-72.
- The term “lipid” is used to describe the components which are used to form lipid mono-, bi-, or multilayers on the surface of the particles (e.g., a core of the particle), that are used in the present invention (e.g., as lipid-coated particles) and may include a PEGylated lipid. Various embodiments provide nanostructures, that are constructed from nanoparticles, which support one or more lipid layers (e.g., bilayer(s) or multilayer(s)). In embodiments according to the present invention, the nanostructures preferably include, for example, a core-shell structure including a porous particle core surrounded by a shell of one or more lipid bilayer(s). In one non-limiting embodiment, the nanostructure (e.g., a porous silica or alum nanostructure) supports the lipid bilayer membrane structure.
- The terms “targeting ligand” and “targeting active species” are used to describe a compound or moiety (e.g., an antigen), which is complexed or covalently bonded to the surface of particle according to the present invention (e.g., either directly on an outer surface of a delivery platform, on an outer lipid layer, or on a supported lipid layer). The targeting ligand, in turn, binds to a moiety on the surface of a cell to be targeted so that the lipid-coated particles may bind to the surface of the targeted cell, enter the cell or an organelle thereof, and/or deposit their contents into the cell. The targeting active species for use in the present invention is preferably a targeting peptide (e.g., a cell penetration peptide, a fusogenic peptide, or an endosomolytic peptide, as otherwise described herein), a polypeptide including an antibody or antibody fragment, an aptamer, or a carbohydrate, among other species that bind to a targeted cell.
- By an “effective amount” or a “sufficient amount” of an agent (e.g., a lipid-coated particle, an antiviral compound, a compound, or an antiviral carrier), as used herein, is that amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, an “effective amount” depends upon the context in which it is being applied. For example, in the context of administering an agent that employs an antiviral compound is, for example, an amount sufficient to achieve decreased viral titer of a virus and/or to treat an infection, as compared to the response obtained without administration of the agent. In another example, in the context of administering an antiviral carrier that employs an antiviral compound is, for example, an amount sufficient to achieve decreased viral titer of a virus and/or to treat an infection, as compared to the response obtained without administration of the antiviral carrier. In yet another example, in the context of administering an antiviral carrier that employs an amount of the antiviral compound is, for example, an amount sufficient to achieve decreased viral titer of a virus and/or to treat an infection, as compared to the response obtained with administration of the amount of the antiviral compound without the antiviral carrier. Thus, an effective amount of an antiviral carrier including an antiviral compound can be compared to any useful control (e.g., an effect determined upon administration of an effective amount of the antiviral compound when used alone, an effect determined upon administration of a buffer, or an effect determined without administration of the antiviral carrier).
- By “subject” is meant a human or non-human animal (e.g., a mammal).
- By “treating” a disease, disorder, or condition in a subject is meant reducing at least one symptom of the disease, disorder, or condition by administrating a therapeutic agent to the subject. By “treating prophylactically” a disease, disorder, or condition in a subject is meant reducing the frequency of occurrence of or reducing the severity of a disease, disorder or condition by administering a therapeutic agent to the subject prior to the onset of disease symptoms. Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; diminishment of extent of disease, disorder, or condition; stabilized (i.e., not worsening) state of disease, disorder, or condition; preventing spread of disease, disorder, or condition; delay or slowing the progress of the disease, disorder, or condition; amelioration or palliation of the disease, disorder, or condition; and remission (whether partial or total), whether detectable or undetectable.
- By “salt” is meant an ionic form of a compound or structure (e.g., any formulas, compounds, or compositions described herein), which includes a cation or anion compound to form an electrically neutral compound or structure. Salts are well known in the art. For example, non-toxic salts are described in Berge S M et al., “Pharmaceutical salts,” J. Pharm. Sci. 1977 January; 66(1):1-19; and in “Handbook of Pharmaceutical Salts: Properties, Selection, and Use,” Wiley-VCH, April 2011 (2nd rev. ed., eds. P. H. Stahl and C. G. Wermuth). The salts can be prepared in situ during the final isolation and purification of the compounds of the invention or separately by reacting the free base group with a suitable organic acid (thereby producing an anionic salt) or by reacting the acid group with a suitable metal or organic salt (thereby producing a cationic salt). Representative anionic salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, camphorate, camphorsulfonate, chloride, citrate, cyclopentanepropionate, digluconate, dihydrochloride, diphosphate, dodecyl sulfate, edetate, ethanesulfonate, fumarate, glucoheptonate, glucomate, glutamate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, hydroxyethanesulfonate, hydroxynaphthoate, iodide, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methylbromide, methylnitrate, methylsulfate, mucate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, theophyllinate, thiocyanate, triethiodide, toluenesulfonate, undecanoate, valerate salts, and the like. Representative cationic salts include metal salts, such as alkali or alkaline earth salts, e.g., barium, calcium (e.g., calcium edetate), lithium, magnesium, potassium, sodium, and the like; other metal salts, such as aluminum, bismuth, iron, and zinc; as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, pyridinium, and the like. Other cationic salts include organic salts, such as chloroprocaine, choline, dibenzylethylenediamine, diethanolamine, ethylenediamine, methylglucamine, and procaine. Exemplary salts include pharmaceutically acceptable salts.
- By “pharmaceutically acceptable salt” is meant a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio.
- By “pharmaceutically acceptable excipient” is meant any ingredient other than a compound or structure (e.g., any formulas, compounds, or compositions described herein) and having the properties of being nontoxic and non-inflammatory in a subject. Exemplary, non-limiting excipients include adjuvants, antiadherents, antioxidants, binders, carriers, coatings, compression aids, diluents, disintegrants, dispersing agents, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), isotonic carriers, lubricants, preservatives, printing inks, solvents, sorbents, stabilizers, suspensing or dispersing agents, surfactants, sweeteners, waters of hydration, or wetting agents. Any of the excipients can be selected from those approved, for example, by the United States Food and Drug Administration or other governmental agency as being acceptable for use in humans or domestic animals. Exemplary excipients include, but are not limited to alcohol, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, glycerol, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactated Ringer's solution, lactose, magnesium stearate, maltitol, maltose, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, Ringer's solution, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium chloride injection, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vegetable oil, vitamin A, vitamin E, vitamin C, water, and xylitol.
- As used herein, the terms “top,” “bottom,” “upper,” “lower,” “above,” and “below” are used to provide a relative relationship between structures. The use of these terms does not indicate or require that a particular structure must be located at a particular location in the apparatus.
- Other features and advantages of the invention will be apparent from the following description and the claims.
-
FIG. 1A-1C shows schematics of exemplary particles. Provided are (A) particular components of an exemplary lipid-coatedparticle 110, including anexemplary core 101 and anexemplary lipid layer 102. Also provided is (B) a schematic of another exemplary lipid-coatedparticle 1100, including anexemplary core 1001 and anexemplary lipid layer 1002. Provide is (C) a schematic for an exemplary loaded, lipid coated-mesoporous silica nanoparticle (LC-MSN), including exemplary components. -
FIG. 2A-2B shows (A) a schematic of ML336 loaded LC-MSN fabrication. The antiviral ML336 was incubated overnight at 4° C. with MSNs at a 10% mass ratio. Liposomes, prepared by combining 77.5% DSPC:2.5% DSPEPEG200:20% cholesterol at mole ratios, were fused to ML336 loaded MSN cores under bath sonication at a 5:1 mass ratio of liposomes:nanoparticles to form ML336 loaded LC-MSNs. Also provided is a schematic (B) of formula (I) and (II) for an exemplary class of quinazolinone-based inhibitors. -
FIG. 3A-3I shows characterization of ML336 LC-MSNs. Provided are (A) TEM images and (B) SEM images of MSNs (all scale bars=50 nm; except (B) bottom left scale bar=100 nm). Also provided are cryo-EM images of (C) ML336 loaded LC-MSNs and (D) unloaded LC-MSNs (scale bar=50 nm). Gray arrows point out examples of the lipid bilayer. Also provided are graphs showing (E) cumulative and (F) percent release (normalized to total ML336 loaded) of ML336 from LC-MSNs. Data represent mean±standard deviation, n=6. Also provided are ultrastructure and pore analysis of mesoporous silica nanoparticles, in which shown are (G,H) SEM images of the hexagonal porous structure of MSNs with different projections. As seen in (G), hexagonal structure is highlighted by the honey comb-like arrangement (A1) and tubular channels (A2). Axis (z) is parallel to the pore's plan. As seen in (H), a tilted image shows tubular channels in a hexagonal arrangement ending by cargo-accessible openings (pore mouth). Provided are (I) graphs of N2 adsorption-desorption isotherm and pore size distribution (inset) for hexagonal small pore MSN. -
FIG. 4A-4F shows (A) DLS measurements of ML336 loaded MSNs and loaded LC-MSNs over the course of a week. ML336 loading and release was determined by comparing sample absorbance values at 320 nm to a standard curve in (B) a lipid solution or (C) a PBS solution. (D) ML336 loading was calculated using the following formula: Total mass loaded=Initial mass of ML336 added −[(mass of ML336 in the supernatant after combination with the lipids)+(mass of ML336 in the supernatant of PBS wash 1)+(mass of ML336 in the supernatant of PBS wash 2)]. Also provided are (E) cumulative and (F) percent release (normalized to total ML336 loaded) of ML336 from LC-MSNs inPBS pH 7,PBS pH 5, and methanol (MeOH). Data represent mean±standard deviation, *=significantly different MeOH group at 18 hours, n=5. -
FIG. 5A-5D shows that ML336 inhibits TC-83 and VEEV (ZPC738 virulent strain) in a dose-dependent manner. Provided are graphs showing (A) TC-83 viral inhibition in HeLa cells at 24 and 48 hours with increasing concentrations of ML336 (n=3); (B) IC-50 of ML336 is 163 nm for HeLa cells at 24 hours of TC-83 infection; (C) VEEV viral inhibition in HeLa cells at 24 hours with increasing concentrations of ML336; and (D) IC-50 of ML336 is 15 nm for HeLa cells at 24 hours of VEEV infection (n=1). -
FIG. 6 shows that ML336 loaded LC-MSNs do not visibly affect cell viability. LIVE (green)/DEAD (red) staining on cells treated with ML336 loaded LC-MSNs, unloaded LC-MSNs, or nothing for 48 hours (scale bar=50 μm; n=3). -
FIG. 7 shows that ML336 loaded LC-MSNs inhibit virus in a dose-dependent manner. *=Significantly different than 25 μg group at same timepoint, #=significantly different than 12.5 μg at the same timepoint, $=significantly different than 6.25 μg group at the same timepoint; p<0.05; data are depicted as mean±standard deviation; n=3. -
FIG. 8A-8C shows that ML336 loaded LC-MSNs inhibited virus in vitro. Provided are (A) phase microscopy images ofcells -
FIG. 9A-9D shows repeatability studies for LC-MSN viral inhibition in vitro. Provided are PFU/mL for loaded, supernatant, pre-released, and untreated groups for (A, C)batch 1 and (B, D)batch 2 in (A,B) study 1 and (C, D)study 2. Note that (A) is also depicted inFIG. 8C . *=Significantly different than loaded group at same timepoint, #=significantly different than supernatant group at the same timepoint, $=significantly different than pre-released group at the same timepoint; p<0.05; data is depicted as mean±standard deviation; n=3 technical replicates and 5 biological replicates. -
FIG. 10A-10D shows LC-MSN cellular internalization by clathrin-mediated endocytosis. (A) LC-MSNs containing a Cy3 dye label were added to inhibitor treated HeLa cells, and the uptake efficiency was visualized using brightfield and fluorescent image overlays (scale bar=25 μm) or (B) quantified through flow cytometry. The inhibitor panel included those targeting pH dependent endocytosis (BAF), clathrin-mediated endocytosis (DYN, CPZ), macropinocytosis (WORT, IPA-3), and caveola-mediated endocytosis (PMA, DYN), while untreated cells with (NI) and without LC-MSN (cells) addition served as controls. (C,D) HeLa cells treated with Cy3 labeled LC-MSNs for 45 min or 20 hr were fixed and stained for microtubules with anti a-tubulin antibodies, actin with phalloidin, and nuclei with DAPI. Confocal images were acquired, and 3D cell images were subjected to isosurface rendering to reveal time-dependent internalization of LC-MSNs (scale bars=10 μmin (C) and 2 μm in (D)). *=Significantly different than no inhibition group; p<0.05; data are depicted as mean±standard deviation, n=3 technical replicates. -
FIG. 11A-11C shows that inhibitors of clathrin-mediated endocytosis reduce both LC-MSN uptake and VSV infection, while inhibitors of caveolae-mediated endocytosis block RVFV and some VSV infection, but not LC-MSN cellular internalization. The inhibitor panel included those targeting pH dependent endocytosis (BAF), clathrin-mediated endocytosis (DYN, CPZ), macropinocytosis (WORT, IPA-3) and caveola-mediated endocytosis (PMA, DYN), while untreated cells with (NI) and without LC-MSN (cells) addition served as controls. (A) HeLa cells were incubated with endocytosis inhibitor treatments for lh prior and during incubation with Cy3 labeled LCMSNs (NP-Cy3), Vesicular Stomatitis virus (VSV), or Rift Valley fever virus (RVFV strain MP-12). At 16 h post nanoparticle or virus addition, cells were washed with PBS twice and prepared for flow cytometry analysis. Representative flow data are shown. No inhibitor (NI) control values were used to normalize the treatment conditions as percent infection for (B) VSV or (C) RVFV (n=2). Overall, these treatment conditions specifically distinguish between common endocytic pathways. -
FIG. 12A-12C shows unloaded LC-MSNs do not affect animal weight in safety studies. (A) Percent weight change in animals dosed with unloaded LC-MSNs or PBS alone over the course of 15 days. (B) Normalized weights of lung, liver, spleen, kidney, and brain to total animal weight in animals dosed with unloaded LC-MSNs or PBS alone over the course of 15 days (data are depicted as mean±standard deviation). Provided are (C) histological analysis of LC-MSN dosed C3H/HeN mice. Mouse tissues were dissected and formalin-fixed onday 15 post-treatment with a vehicle control (PBS) or LC-MSNs at 0.11 g LC-MSNs/kg/day for four days. Histological specimens were prepared through paraffin embedding and sectioning, followed by hematoxylin and eosin staining. Three animals per group were analyzed and representative images are shown. In the brain, the outer cortex is shown and displayed no obvious differences between the LC-MSN dosed and control groups. Similarly, the spleen and kidney sections exhibited normal morphology without signs of toxicity. In some samples, granulomas that contained collections of macrophages embedded in the lung and liver (indicated by asterisks) were identified and indicative of very mild symptoms. -
FIG. 13A-13C shows ML336 loaded LC-MSNs show reduction of viral load in vivo. Provided are (A) a survival curve for TC-83 infected animals treated with 1 mg ML336 loaded LC-MSNs twice a day for 4 days, as well as viral load in (B) brain and (C) spleen normalized to organ mass after 4 days of infection and treatment with 1.5 mg ML336 loaded LC-MSNs (*=significantly different from PBS group; p<0.05; data are depicted as mean±standard deviation). -
FIG. 14A-14C shows viral titer in tissues of TC-83 infected mice. Provided are the viral loads in (A) serum, (B) kidney, and (C) liver atday 4 post-infection via intranasal challenge of C3H/HeN mice with VEEV strain TC-83 were measured by standard plaque assays normalized to volume (ml) or organs mass (gram). Viral loads from four treatment conditions are shown for ML-336 loaded LC-MSN (circle), unloaded LC-MSN (square), free ML-336 (triangle), and vehicle only (PBS) (upside down triangle) with mean from 5 samples per condition. The limit of detection (LOD) is 100PFU, and samples at or below this threshold are all listed at LOD. - The present invention relates to the use of lipid-coated particles to deliver compounds having reduced stability and/or solubility. In one instance, ML336 is a small molecule inhibitor that displays antiviral activity but has poor stability and solubility characteristics. Here, we show that ML336-loaded lipid coated mesoporous silica nanoparticles (LC-MSNs) possess good colloidal stability. Such particles ML336-loaded LC-MSNs inhibited Venezuelan equine encephalitis virus (VEEV) in vitro in a dose-dependent manner, as compared to untreated controls. In vivo safety studies were conducted in C3H/HeN mice, and LC-MSNs were not toxic when at tested doses. Furthermore, ML336-loaded LC-MSNs showed significant reduction in brain viral titer in VEEV TC-83 infected mice, as compared to PBS treated controls. Overall, these results highlight the utility of LC-MSNs as drug delivery vehicles to treat VEEV infections.
-
FIG. 1A provides an exemplary lipid-coatedparticle 110 including aninner core 101 and an outer lipid layer 102 (e.g., a lipid bilayer, a multilamellar lipid layer, etc.) disposed around thecore 101. In some non-limiting instances, the core is porous (e.g., including a plurality of cores). In other non-limiting instances, the lipid layer can include any useful lipid (e.g., a PEGylated lipid), useful component (e.g., a cholesterol), and/or useful targeting ligand (e.g., any described herein). Furthermore, the outer lipid layer can include a plurality of layers, in which each layer can be a lipid bilayer. In this manner, the layer can be multilamellar because it includes multiple lamellae (or multiple layers). -
FIG. 1B provided another an exemplary lipid-coatedparticle 1100 including aninner core 1001 and an outer lipid layer 1002 (e.g., a lipid bilayer, a multilamellar lipid layer, etc.) disposed around thecore 1001. As can be seen, the core can have any useful features or characteristics. In one embodiment, the core is a monosized (e.g., polydispersity index<0.1) particle. The core can have any useful shape, morphology, pore size, and pore distribution. - The lipid-coated particle can be characterized by any useful manner either before loading of cargo or after loading of cargo (e.g., overall charge, dimension, dispersity, etc.). Furthermore, components of the particle (e.g., the core or the lipid layer) can also be characterized by any useful manner (e.g., pore size, core size, core charge, lipid layer thickness, lipid layer charge, etc.).
- Cargo (e.g., a compound) can be loaded in any useful manner. In one instance, cargo is introduced to the core, and then the loaded core is exposed to a solution containing liposomes, which results in the formation of a lipid layer disposed around the loaded core.
- Compounds, Including Antiviral Compounds
- The present invention can include the any useful compound (e.g., an antiviral compound). In one instance, the compound has reduced stability and/or reduced solubility, thereby would benefit from the use of a carrier (e.g., any described herein). In yet other embodiments, the compound is hydrophobic (e.g., determined in any useful manner, such as any herein).
- In some embodiments, the compound has limited aqueous solubility (e.g., from about 20 μg/mL to about 150 μg/mL, such as from 20 μg/mL to 50 μg/mL, 20 μg/mL to 100 μg/mL, 20 μg/mL to 150 μg/mL, 50 μg/mL to 100 μg/mL, or 50 μg/mL to 150 μg/mL) in an aqueous solvent. Solubility can be determined in any useful manner, such as an automated kinetic solubility method at any useful temperature (e.g., of from about 20° C. to 30° C., such as about 23° C.). In one embodiment of such a solubility method, a saturated pH-buffered aqueous solution is prepared, and the concentration of the compound is determined analytically (e.g., by gas chromatography, UV absorbance, liquid chromatography mass spectrometry (LC-MS), etc.), and the obtained spectrum is compared to a control spectrum for a precipitation-free reference solution. Exemplary aqueous solvents include phosphate buffered saline (PBS, 137 mM NaCl, 2.7 mM KCl, 10 mM sodium phosphate dibasic, 2 mM potassium phosphate monobasic and a pH of 7.4) or a cell medium (e.g., a cytopathic effect (CPE) medium including high glucose DMEM (Dulbecco's Modified Eagle's Medium) with 10% fetal bovine serum and 1× penicillin streptomycin solution (Pen/Strep)).
- In some embodiments, the compound has limited aqueous stability (e.g., of about 80% or less of a remaining amount of the compound after incubating in mouse plasma for about 3 hours; from about 20% to about 80% remaining amount of the compound after incubating in mouse plasma for about 3 hours; or from about 20% to 90% remaining amount of the compound after incubating at PBS, pH 7.4). Stability can be determined in any useful manner. In one instance, the compound is dissolved in a solvent (e.g., at 10 μM in PBS at pH 7.4 with 1% dimethylsulfoxide (DMSO), human plasma, or mouse plasma) and analyzed at various timepoints (e.g., numerous timepoints from 0 to 48 hours). The concentration of the compound is determined analytically (e.g., by gas chromatography, UV absorbance, LC-MS, etc.), and absolute areas under the curve can be employed at each time point to determine the relative percent of the remaining parent compound. For experiments including plasma, stability can be determined at about 37° C. with optional incubation at these temperature, centrifugation, and shaking.
- In some embodiments, the compound has a computed hydrophobicity XLogP3-AA of from about 2 to about 10 (e.g., from 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 20, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, and 9 to 10). Hydrophobicity values can be determined in any useful manner, e.g., XLogP3-AA, which is a computationally generated octanol-water partition coefficient or distribution coefficient including an additive model, e.g., that can be determined according to Cheng T et al., “Computation of octanol-water partition coefficients by guiding an additive model with knowledge,” J. Chem. Inf. Model. 2007; 47:2140-8; ACD/LogP, which is another computationally generated octanol-water partition coefficient or distribution coefficient having correction factors, e.g., that can be determined according to Petrauskas A A et al., “ACD/Log P method description,” Perspect. Drug Discovery Des. 2000; 19:99-116 and/or Walker M J, “Training ACD/LogP with experimental data,” QSAR Comb. Sci. 2004; 23:515-20; and/or topological polar surface area (TPSA), which can be computed using an algorithm according to Ertl P et al., “Fast calculation of molecular polar surface area as a sum of fragment-based contributions and its application to the prediction of drug transport properties,” J. Med. Chem. 2000; 43:3714-7, each of which is incorporated herein by reference in its entirety. Non-limiting, exemplary antiviral compounds include, e.g., (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-nitro-N-phenylbenzamide; (E)-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-phenylbenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(4-methoxyphenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(2-fluorophenyl)-5-nitrobenzamide; (E)-4-chloro-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-phenylbenzamide; (E)-2-((1-ethyl-4-methylpiperazin-2-ylidene)amino)-5-nitro-N-phenylbenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-fluoro-N-phenylbenzamide; (E)-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(2-fluorophenyl)benzamide; (E)-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(3-fluorophenyl)-benzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(4-methoxyphenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(3-fluorophenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-phenyl-5-(trifluoromethyl)benzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(4-fluorophenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(2-methoxyphenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-(3-methoxyphenyl)-5-nitrobenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-isopropyl-5-nitrobenzamide; (E)-N-benzyl-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-nitrobenzamide; (E)-4-((1,4-dimethylpiperazin-2-ylidene)amino)-N-phenylpyridazine-3-carboxamide; (E)-methyl 4-((1,4-dimethylpiperazin-2-ylidene)amino)-3-(phenylcarbamoyl)benzoate; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-nitro-N-(thiophen-3-yl)benzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-4,5-difluoro-N-phenylbenzamide; (E)-5-cyano-2-((1,4-dimethylpiperazin-2-ylidene)amino)-4-fluoro-N-phenylbenzamide; (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-N-methyl-5-nitrobenzamide; 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-5-nitro-N-phenylbenzamide (ML336,)<LogP3-AA of 2.1, cLogP of 3.4, Topological Polar Surface Area (TPSA) of 93.8 Å2); 2-[(4-ethylpiperazin-1-yl)methyl]-3-(2-fluorophenyl)-6-nitroquinazolin-4-one (XLogP3-AA of 2.6, TPSA of 85 Å2); 6,7-difluoro-3-phenyl-2-(piperazin-1-ylmethyl)quinazolin-4-one (XLogP3-AA of 2, TPSA of 47.9 Å2); 2-(1,3-diazinan-1-ylmethyl)-6-nitro-3-phenylquinazolin-4-one (XLogP3-AA of 2.1, TPSA of 93.8 Å2) 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-N-(2-fluorophenyl)-5-nitrobenzamide (XLogP3-AA of 2.2, TPSA of 93.8 Å2); 5-cyano-2-[(1,4-dimethylpiperazin-2-ylidene)amino]-N-phenylbenzamide (XLogP3-AA of 2, TPSA of 71.7 Å2); 2-(N-methyl-4-phenylmethoxyanilino)pyrido[3,2-e][1,3]thiazin-4-one (XLogP3-AA of 4.1, TPSA of 80.1 Å2); 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-N-(4-methoxyphenyl)-5-nitrobenzamide (XLogP3-AA of 2, TPSA of 103 Å2); 2-[(1,4-dimethylpiperazin-2-ylidene)amino]-N-(2-fluorophenyl)-5-nitrobenzamide (XLogP3-AA of 2.2, TPSA of 93.8 Å2); N-[1-[2-[2-chloro-6-(4-chlorophenoxy)pyridin-4-yl]-4-methyl-1,3-thiazol-5-yl]ethylideneamino]aniline (A5,XLogP3-AA of 7.6, TPSA of 87.6 Å2); N-(1,3-benzothiazol-2-yl)-N-methyl-2-thiophen-2-ylacetamide; N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2-yl)-2-thiophen-2-ylacetamide (XLogP3-AA of 2.9, TPSA of 98.5 Å2); N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2-yl)-3-fluoro-2-thiophen-2-ylacetamide; N-phenyl-N-(4,5,6,7-tetrahydro-1,3-benzothiazol-2-yl)-2-thiophen-2-ylacetamide (XLogP3-AA of 4.6, TPSA of 89.7 Å2); 2-[(4-ethylpiperazin-1-yl)methyl]-3-(2-fluorophenyl)-6-nitroquinazolin-4-one (XLogP3-AA of 2.6, TPSA of 85 Å2); 2-(4-phenylmethoxyanilino)pyrido[3,2-e][1,3]thiazin-4-one (XLogP3-AA of 3.9, TPSA of 88.9 Å2); N-[4-[[4-[9-[4-[(4-acetamidophenyl)sulfonylamino]phenyl]fluoren-9-yl]phenyl]sulfamoyl]phenyl]acetamide (compound AN-329/40863801,XLogP3-AA of 6.1, TPSA of 167 Å2); N-[(Z)-1-[4-(dimethylamino)phenyl]-3-(3-imidazol-1-ylpropylamino)-3-oxoprop-1-en-2-yl]benzamide (compound 1111684,XLogP3-AA of 2.9, TPSA of 79.3 Å2); (6Z)-3-[2-[4-(2-methoxyphenyl)piperazin-1-yl]ethoxy]-6-(4-phenyl-1,2-dihydropyrazol -3-ylidene)cyclohexa-2,4-dien-1-one (compound 6052346,XLogP3-AA of 4.2, TPSA of 66.1 Å2); 9H-fluoren-9-ylmethyl N-1-[[1-[[1-(2-((1-carboxy-3-methylbutyl)carboxamido) pyrrolidinyl)H2-(t-butoxy)methyl]-1-oxoethan-2-yl]amino]-5-[[amino-[(4-methoxy-2,3,6-trimethylphenyl)sulfonylamino]methylidene]amino]-1-oxopentan-2-yl]carbamate (compound JFD02946); rintatolimod (Ampligen®); 4-(6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-7-ylmethyl)-2-(5-methylfuran-2-yl)-1,3-thiazole (Z1139230991,XLogP3-AA of 1.8, TPSA of 83.3 Å2); 2-(furan-2-yl)-4-[(2-methyl-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-7-yl)methyl]-1,3-thiazole (Z1139583813, XLogP3-AA of 1.8, TPSA of 83.3 Å2); 3-(3,5-dimethyl-1,2-oxazol-4-yl)-N-[4-(2-oxo-1,3-dihydroindol-5-yl)-1,3-thiazol-2-yl]propenamide (Z70927013,XLogP3-AA of 2.1, TPSA of 125 Å2); 4-ethyl-N-(3-methoxypropyl)-7,8,9,10-tetrahydro-4H-[1]benzothieno[3,2-f]pyrrolo[1,2-a][1,4]diazepine-5(6H)-carboxamide (G281-1564, LogP of 3.6, ACD/LogP of 4.85, 74.74 Å2); N-(3-ethoxypropyl)-4-isopropyl-7,8,9,10-tetrahydro-4H-[1]benzothieno[3,2-f]pyrrolo[1,2-a][1,4]diazepine-5(6H)-carboxamide (G281-1485, LogP of 4.276, ACD/Log P of 5.73, Polar Surface Area (PSA) of 74.74 Å2), as well as salts thereof. Further antiviral compounds are disclosed in U.S. Pat. No., 9,580,393; U.S. Pat. Pub. No. 2013/085133; Chung D H et al., “Discovery of a broad-spectrum antiviral compound that inhibits pyrimidine biosynthesis and establishes a
type 1 interferon-independent antiviral state,” Antimicrob. Agents Chemother. 2016; 60(8):4552-62; Schecter S et al., “Novel inhibitors targeting Venezuelan equine encephalitis virus capsid protein identified using In Silico Structure-Based-Drug-Design,” Sci. Rep. 2017; 7:art. no. 17705 (16 pp.); Thomas D R et al., “Identification of novel antivirals inhibiting recognition of Venezuelan equine encephalitis virus capsid protein by the importin α/β1 heterodimer through high-throughput screening,” Antiviral Res. 2018; 151:8-19; Chung D H et al., “Discovery of a novel compound with anti-Venezuelan equine encephalitis virus activity that targets the nonstructural protein 2,” PLoS Pathog. 2014; 10(6):e1004213; Julander J G et al., “Treatment of Venezuelan equine encephalitis virus infection with (−)-carbodine,” Antiviral Res. 2008; 80(3):309-15; Julander J G et al., “C3H/HeN mouse model for the evaluation of antiviral agents for the treatment of Venezuelan equine encephalitis virus infection,” Antiviral Res. 2008; 78(3):230-41; Kehn-Hall K et al., “Modulation of GSK-3beta activity in Venezuelan equine encephalitis virus infection,” PLoS One 2012; 7(4):e34761; Langsjoen R M et al., “Host oxidative folding pathways offer novel anti-chikungunya virus drug targets with broad spectrum potential,” Antiviral Res. 2017; 143:246-51; and Madsen C et al., “Small molecule inhibitors of Ago2 decrease Venezuelan equine encephalitis virus replication,” Antiviral Res. 2014; 112:26-37, each of which is incorporated herein by reference in its entirety. - In yet other embodiments, the compound or antiviral compound is from a class of quinazolinone-based inhibitors. In particular embodiments, the compound has a structure of formula (I), (II), or (III), or a salt thereof (
FIG. 2B ). In some embodiments, R2 includes any substituent including an aryl or heterocyclyl moiety. In particular embodiments, each R2 is, independently, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, or optionally substituted alkheterocyclyl. - In some embodiments, each of R1, R3, R4, R5, R7, and R8 is any useful substituent (e.g., any described herein, such as H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, optionally substituted alkheterocyclyl, halo, nitro, amino, azido, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, optionally substituted cycloalkyl, or optionally substituted spirocyclyl for two nearby R groups taken together). In particular embodiments, each R1 is, independently, H or optionally substituted alkyl. In some embodiments, each R2 is, independently, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkaryl, or optionally substituted alkheterocyclyl. In other embodiments, each R4, R5, R7, and R8 is, independently, H, optionally substituted alkyl, halo, nitro, amino, azido, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl, or in which R4 and R5, taken together, or R7 and R8, taken together, form an optionally substituted spirocyclyl. In some embodiments, each R3 and R6 is, independently, H, optionally substituted alkyl, halo, nitro, nitroso, amino, azido, carboxyl, cyano, hydroxyl, optionally substituted hydroxyalkyl, optionally substituted haloalkyl, optionally substituted perfluoroalkyl, or optionally substituted cycloalkyl.
- Other exemplary cargos include an acidic, basic, and hydrophobic drug (e.g., antiviral agents, antibiotic agents, etc.); a protein (e.g., antibodies, carbohydrates, etc.); a nucleic acid (e.g., DNA, RNA, small interfering RNA (siRNA), minicircle DNA (mcDNA), small hairpin RNA (shRNA), complementary DNA (cDNA), naked DNA, and plasmid, as well as chimeras, single-stranded forms, duplex forms, and multiplex forms thereof and including nucleic acid sequences encoding any of these and including one or more modified nucleic acids); a CRISPR component, a nuclease, a plasmid, a plasmid that encodes a CRISPR component, a ribonucleoprotein complex, a Cas enzyme or an ortholog or homolog thereof, a guide RNA, as well as a nucleic acid sequence encoding any of these or a complement thereof); a diagnostic/contrast agent, like quantum dots, iron oxide nanoparticles, gadolinium, and indium-111; a small molecule; a carbohydrate; a drug, a pro-drug, a vitamin, an antibody, a protein, a hormone, a growth factor, a cytokine, a steroid, an anticancer agent, a fungicide, an antimicrobial, an antibiotic, an antiviral agent, etc.; a morphogen; a toxin, e.g., a bacterial protein toxin; a peptide, e.g., an antimicrobial peptide; an antigen; an antibody; a detection agent (e.g., a particle, such as a conductive particle, a microparticle, a nanoparticle, a quantum dot, a latex bead, a colloidal particle, a magnetic particle, a fluorescent particle, etc.; or a dye, such as a fluorescent dye, a luminescent dye, a chemiluminescent dye, a colorimetric dye, a radioactive agent, an electroactive detection agent, etc.); a label (e.g., a quantum dot, a nanoparticle, a microparticle, a barcode, a fluorescent label, a colorimetric label, a radio label (e.g., an RF label or barcode), avidin, biotin, a tag, a dye, a marker, an electroactive label, an electrocatalytic label, and/or an enzyme that can optionally include one or more linking agents and/or one or more dyes); a capture agent (e.g., such as a protein that binds to or detects one or more markers (e.g., an antibody or an enzyme), a globulin protein (e.g., bovine serum albumin), a nanoparticle, a microparticle, a sandwich assay reagent, a catalyst (e.g., that reacts with one or more markers), and/or an enzyme (e.g., that reacts with one or more markers, such as any described herein)); as well as combinations thereof.
- Core
- The present invention relates, in part, to a particle having a core. The core can provide any useful benefit. In particular non-limiting embodiments, the core provides a surface upon which a lipid layer can be supported. In other non-limiting embodiments, the core provides a charged surface that allows for electrostatic interactions with the cargo and/or the lipid layer, or a portion thereof.
- The core can be characterized in any useful manner. In one instance, the core can be characterized by a first dimension (e.g., core circumference, pore size of the core, core diameter, core length, or core width). Exemplary values for a core dimension (e.g., core circumference, core diameter, core length, or core width, as well as an average or mean value for any of these) include, without limitation, greater than about 1 nm (e.g., greater than about 5 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 125 nm, 150 nm, 200 nm, 300 nm, 500 nm, 750 nm, 1 μm, 2 μm, 5 μm, 10 μm, 20 μm, or more), including of from about 5 nm to about 300 nm (e.g., from 5 nm to 20 nm, 5 nm to 30 nm, 5 nm to 40 nm, 5 nm to 50 nm, 5 nm to 75 nm, 5 nm to 100 nm, 5 nm to 150 nm, 5 nm to 200 nm, 5 nm to 250 nm, 10 nm to 20 nm, 10 nm to 30 nm, 10 nm to 40 nm, 10 nm to 50 nm, 10 nm to 75 nm, 10 nm to 100 nm, 10 nm to 150 nm, 10 nm to 200 nm, 10 nm to 250 nm, 10 nm to 300 nm, 25 nm to 30 nm, 25 nm to 40 nm, 25 nm to 50 nm, 25 nm to 75 nm, 25 nm to 100 nm, 25 nm to 150 nm, 25 nm to 200 nm, 25 nm to 250 nm, 25 nm to 300 nm, 50 nm to 75 nm, 50 nm to 100 nm, 50 nm to 150 nm, 50 nm to 200 nm, 50 nm to 250 nm, 50 nm to 300 nm, 75 nm to 100 nm, 75 nm to 150 nm, 75 nm to 200 nm, 75 nm to 250 nm, 75 nm to 300 nm, 100 nm to 150 nm, 100 nm to 200 nm, 100 nm to 250 nm, 100 nm to 300 nm, 150 nm to 200 nm, 150 nm to 250 nm, 150 nm to 300 nm, 200 nm to 250 nm, 200 nm to 300 nm, 250 nm to 300 nm, or 275 nm to 300 nm). In one instance, the particle includes a porous core (e.g., a silica core that is spherical and ranges in diameter from about 10 nm to about 250 nm (e.g., having a mean diameter of about 150 nm)). In particular embodiments, the silica core is monodisperse or polydisperse in size distribution.
- The core can be further characterized by an electrostatic property. In some embodiments, the core has a negative charge (e.g., a net negative charge), such as a zeta potential of from about −10 mV to about −200 mV (e.g., from −10 mV to −100 mV, −10 mV to −75 mV, −10 mV to −50 mV, −10 mV to −30 mV, −15 mV to −100 mV, −15 mV to −75 mV, −15 mV to −50 mV, −15 mV to −30 mV, −20 mV to −100 mV, −20 mV to −75 mV, −20 mV to −50 mV, −20 mV to −30 mV, −30 mV to −100 mV, −30 mV to −75 mV, −30 mV to −50 mV, −40 mV to −100 mV, −40 mV to −75 mV, −40 mV to −50 mV, −50 mV to −100 mV, −50 mV to −75 mV, −60 mV to −100 mV, or −60 mV to −75 mV).
- The core can be porous. In particular embodiments, the pore has a dimension (e.g., average pore size, pore diameter, pore radius, pore circumference, pore length, pore width, or pore depth) that is greater than about 0.5 nm (e.g., of from about 0.5 nm to about 30 nm, including from 0.5 nm to 10 nm, 0.5 nm to 20 nm, 0.5 nm to 25 nm, 1 nm to 10 nm, 1 nm to 15 nm, 1 nm to 20 nm, 1 nm to 25 nm, 1 nm to 30 nm, 2 nm to 5 nm, 2 nm to 10 nm, 2 nm to 20 nm, 2 nm to 25 nm, or 2 nm to 30 nm).
- A particle or a portion thereof (e.g., a core) may have a variety of shapes and cross-sectional geometries that may depend, in part, upon the process used to produce the particles. The core or particle can be a nanoparticle (e.g., having a diameter less than about 1 μm) or a microparticle (e.g., having a diameter greater than or equal to about 1 μm). In one embodiment, a core or particle may have a shape that is a sphere, a donut (toroidal), a rod, a tube, a flake, a fiber, a plate, a wire, a cube, or a whisker. A collection of cores may have two or more of the aforementioned shapes. In one embodiment, a cross-sectional geometry of the core may be one or more of circular, ellipsoidal, triangular, rectangular, or polygonal. In one embodiment, a core may consist essentially of non-spherical cores. For example, such cores may have the form of ellipsoids, which may have all three principal axes of differing lengths, or may be oblate or prelate ellipsoids of revolution. Non-spherical cores alternatively may be laminar in form, wherein laminar refers to particles in which the maximum dimension along one axis is substantially less than the maximum dimension along each of the other two axes. Non-spherical cores may also have the shape of frusta of pyramids or cones, or of elongated rods. In one embodiment, the cores may be irregular in shape. In one embodiment, a plurality of cores may consist essentially of spherical cores. Particles and cores for use in the present invention may be PEGylated and/or aminated as otherwise described in Int. Pub. Nos. WO 2015/042268 and WO 2015/042279, which is incorporated herein by reference in their entirety.
- The particle size distribution (e.g., size of the core for the particle or a size of the silica carrier), according to the present invention, depends on the application, but is principally monodisperse (e.g., a uniform sized population varying no more than about 5-20% in diameter, as otherwise described herein). In certain embodiments, particles or cores can range, e.g., from around 1 nm to around 500 nm in size, including all integers and ranges there between. The size is measured as the longest axis of the core. In various embodiments, the cores are from around 5 nm to around 500 nm and from around 10 nm to around 100 nm in size. In certain alternative embodiments, the cores or particles are monodisperse and range in size from about 25 nm to about 300 nm. The sizes used preferably include 50 nm (+/−10 nm) and 150 nm (+/−15 nm), within a narrow monodisperse range, but may be more narrow in range.
- When the core is porous, the pores can be from around 0.5 nm to about 25 nm in diameter, often about 1 to around 20 nm in diameter, including all integers and ranges there between. In one embodiment, the pores are from around 1 to around 10 nm in diameter. In one embodiment, around 90% of the pores are from around 1 to around 20 nm in diameter. In another embodiment, around 95% of the pores are around 1 to around 20 nm in diameter.
- In certain embodiments, preferred cores or particles according to the present invention: are monodisperse and range in size from about 25 nm to about 300 nm; exhibit stability (colloidal stability); have single cell binding specification to the substantial exclusion of non-targeted cells; are anionic, neutral or cationic for specific targeting (preferably cationic); are optionally modified with agents such as PEI (polyethylene imine), NMe3+, dye, crosslinker, ligands (ligands provide neutral charge); and optionally, are used in combination with a cargo to be delivered to the target.
- In certain alternative embodiments, the present invention is directed to cores or particles of a particular size (diameter) ranging from about 0.5 to about 30 nm, about 1 nm to about 30 nm, often about 5 nm to about 25 nm (preferably, less than about 25 nm), often about 10 to about 20 nm, for administration in any useful route. In some embodiments, these cores or particles are often monodisperse and provide colloidally stable compositions. These compositions can be used to target host cells because of enhanced biodistribution/bioavailability of these compositions, and optionally, specific cells, with a wide variety of therapeutic and/or diagnostic agents that exhibit varying release rates at the site of activity.
- The cores can be produced in any useful manner. In one instance, cores are formed by templating with a surfactant, a cross-linked micelle, a detergent, or any other useful molecule (see, e.g., Gao F et al., J. Phys. Chem. B. 2009; 113:1796-804; Lin Y S et al., Chem. Mater. 2009; 21(17):3979-86; and Zhang K et al., J. Am. Chem. Soc. 2013 Feb. 20; 135(7):2427-30). In yet another instance, cores are formed by dendritic growth (see, e.g., Shen D et al., Nano Lett. 2014 Feb. 12; 14(2):923-32). Each batch of cores or particles can be characterized in any useful manner, such as by assessment of size and surface charge using dynamic light scattering (DLS) (NIST-NCL PCC-1 and PCC-2) and electron microscopy (NIST-NCL PCC-7 and PCC-15) and verification of low endotoxin contamination per health industry product standards (NCL STE-1.1). Resultant cores can be further processed, such as by modifying core condensation (e.g., by using acidified ethanol for silica), modifying core surface charge (e.g., by use of amine-containing silanes, such as APTES), etc.
- The core can be formed of any useful material (e.g., a metal oxide, alum, silica, including mesoporous forms thereof). In particular embodiments, the core is composed of a mesoporous silica nanoparticle (MSN). Exemplary, non-limiting MSNs for use in the present invention are described in Int. Pub. Nos. WO 2015/042268 and WO 2015/042279, each of which is incorporated herein in its entirety.
- Lipid layer
- The present invention relates to a lipid layer disposed around a core. The lipid layer can be characterized in any useful manner, such as by the thickness of the layer (e.g., of from about 5 nm to about 50 nm), the number of layers within the lipid layer (e.g., two, three, four, five, six, seven, or more lipid bilayers), and/or the net charge of the lipid layer (e.g., a net non-negative charge, such as a net positive charge; or as determined by the composition of the lipid layer, such as a layer formed by use of a liposome formulation having more than about 20 mol. % of a cationic lipid, such as any herein (e.g., DOTAP)).
- The lipid layer can include any useful component, including a cationic lipid, a pegylated lipid, a zwitterionic lipid, and/or a cholesterol. For instance, the lipid layer can include any useful lipid or combination of lipids or component, such as one or more lipids selected from the group of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-[phosphor-L-serine] (DOPS), 1,2-dioleoyl-3-trimethylammonium-propane (18:1 DOTAP), 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DOPG), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (18:1 PEG-2000 PE), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (16:0 PEG-2000 PE), 1-oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]lauroyl]-sn-glycero-3-phosphocholine (18:1-12:0 NBD PC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy-(polyethylene glycol)-2000] (DSPE-PEG2000), 1-palmitoyl-2-{12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]lauroyl}-sn-glycero-3-phosphocholine (16:0-12:0 NBD PC), a sterol (e.g., cholesterol, desmosterol, diplopterol, cholestanol, cholic acid, 12-deoxycholic acid, 7-deoxycholic acid, or a derivative thereof, such as cholesterol sulfate), and mixtures thereof and conjugated forms thereof (e.g., conjugated to PEG moieties, peptides, polypeptides, including immunogenic peptides, proteins and antibodies, and nucleic acids (e.g., RNA and DNA) by way of a covalent bond or by way of any useful linker (e.g., any described herein).
- Exemplary, non-limiting sterols include cholesterol (e.g., from ovine wool or from plant sources), campestanol, campesterol, cholestanol, cholestenone, desmosterol, 7-dehydrodesmosterol, dehydroepiandrosterone (DHEA), desmosterol, diosgenin, FF-MAS (14-demethyl-14-dehydrolanosterol), lanosterol, lathosterol, pregnenolone, sitostanol, sitosterol, stigmasterol, zymosterol, zymostenol, zymosterone, as well as derivatives thereof, such as sulfates thereof, esters thereof, stereoisomers thereof, deuterated forms thereof, sulfonated forms thereof, phosphorylated forms thereof, unsaturated forms thereof, keto forms thereof, oxidized forms thereof, an oxysterol thereof, PEGylated forms thereof (e.g., cholesterol-(polyethylene glycol-600)), or substituted forms thereof (e.g., having one or more hydroxyl, epoxy, alkyl, phospho, and/or halo, such as fluoro).
- Cores, lipids, and cargos can be PEGylated with a variety of polyethylene glycol-containing compositions as described herein. PEG molecules can have a variety of lengths and molecular weights and include, but are not limited to,
PEG 200,PEG 1000, PEG 1500,PEG 2000, PEG 4600, PEG 5000, PEG 10,000, PEG-peptide conjugates or combinations thereof. - In one instance, the lipid layer includes DOPE and DOTAP. In another instance, the lipid layer includes a zwitterionic lipid (e.g., DOPC, DPPC, DOPE, DPPE, DSPE, DLPC, DMPC, POPC, or SOPC) with an optional PEG (e.g., PEG, PEG-2000 PE, PEG conjugated to DOPE, PEG conjugated to DPPE, PEG conjugated to DSPE, etc.).
- In yet another instance, the lipid layer includes DOTAP and cholesterol in a 1:1 molar ratio. In another instance, the lipid layer includes PEG. In yet another instance, the lipid layer includes DOPE. In one instance, the lipid layer includes DOTAP in combination with about 4 mol. % DOPE, about 47 mol. % cholesterol, and about 2 mol. % DSPE-PEth000. In another instance, the lipid layer includes about 10 to about 50 mol. % DOTAP, about 40 to 50 mol. % cholesterol, about 0 to 40 mol. % DOPE, and about 1 to 5 mol. % of a PEGylated lipid.
- The lipid layer can be formed by employing any useful lipid formulation. A non-limiting exemplary formulation can include the following: about 1 mol. % to about 5 mol. % of a PEGylated lipid (e.g., from 1 mol. % to 3 mol. %, 1 mol. % to 4 mol. %, 2 mol. % to 3 mol. %, 2 mol. % to 4 mol. %, 2 mol. % to 5 mol. %, 3 mol. % to 4 mol. %, or 3 mol. % to 5 mol. %); about 30 mol. % to about 60 mol. % of a sterol (e.g., from 30 mol. % to 50 mol. %, 35 mol. % to 50 mol. %, 35 mol. % to 60 mol. %, 40 mol. % to 50 mol. %, 40 mol. % to 60 mol. %, 45 mol. % to 50 mol. %, 45 mol. % to 60 mol. %, 50 mol. % to 60 mol. %, or 55 mol. % to 60 mol. %); about 20 mol. % to about 90 mol. % of a cationic lipid (e.g., from 20 mol. % to 30 mol. %, 20 mol. % to 40 mol. %, 20 mol. % to 50 mol. %, 20 mol. % to 60 mol. %, 20 mol. % to 70 mol. %, 20 mol. % to 80 mol. %, 30 mol. % to 40 mol. %, 30 mol. % to 50 mol. %, 30 mol. % to 60 mol. %, 30 mol. % to 70 mol. %, 30 mol. % to 80 mol. %, 30 mol. % to 90 mol. %, 40 mol. % to 50 mol. %, 40 mol. % to 60 mol. %, 40 mol. % to 70 mol. %, 40 mol. % to 80 mol. %, 40 mol. % to 90 mol. %, 50 mol. % to 60 mol. %, 50 mol. % to 70 mol. %, 50 mol. % to 80 mol. %, 50 mol. % to 90 mol. %, 60 mol. % to 70 mol. %, 60 mol. % to 80 mol. %, 60 mol. % to 90 mol. %, 70 mol. % to 80 mol. %, 70 mol. % to 90 mol. %, or 80 mol. % to 90 mol. %); and about 0 mol. % to about 40 mol. % of a zwitterionic lipid (e.g., 0 mol. % to 3 mol. %, 0 mol. % to 5 mol. %, 0 mol. % to 7 mol. %, 0 mol. % to 10 mol. %, 0 mol. % to 15 mol. %, 0 mol. % to 20 mol. %, 0 mol. % to 25 mol. %, 0 mol. % to 30 mol. %, 0 mol. % to 35 mol. %, 3 mol. % to 5 mol. %, 3 mol. % to 7 mol. %, 3 mol. % to 10 mol. %, 3 mol. % to 15 mol. %, 3 mol. % to 20 mol. %, 3 mol. % to 25 mol. %, 3 mol. % to 30 mol. %, 3 mol. % to 35 mol. %, 3 mol. % to 40 mol. %, 7 mol. % to 10 mol. %, 7 mol. % to 15 mol. %, 7 mol. % to 20 mol. %, 7 mol. % to 25 mol. %, 7 mol. % to 30 mol. %, 7 mol. % to 35 mol. %, 73 mol. % to 40 mol. %, 10 mol. % to 15 mol. %, 10 mol. % to 20 mol. %, 10 mol. % to 25 mol. %, 10 mol. % to 30 mol. %, 10 mol. % to 35 mol. %, 10 mol. % to 40 mol. %, 15 mol. % to 20 mol. %, 15 mol. % to 25 mol. %, 15 mol. % to 30 mol. %, 15 mol. % to 35 mol. %, 15 mol. % to 40 mol. %, 20 mol. % to 25 mol. %, 20 mol. % to 30 mol. %, 20 mol. % to 35 mol. %, 20 mol. % to 40 mol. %, 25 mol. % to 30 mol. %, 25 mol. % to 35 mol. %, 25 mol. % to 40 mol. %, 30 mol. % to 35 mol. %, 30 mol. % to 40 mol. %, or 35 mol. % to 40 mol. %), or salts of any of these (e.g., pharmaceutically acceptable salts, such as any described herein).
- In particular embodiments, the ratio of the sterol to the cationic lipid is about 1:1. In other embodiments, the lipid formulation includes about 2% of the PEGylated lipid. In yet other embodiments, the lipid formulation includes about 30 mol. % to about 60 mol. % of the cationic lipid.
- The lipid formulation can include any useful lipid or component. Exemplary PEGylated lipids (e.g., a lipid having a poly(ethylene glycol moiety)) include PEGylated DSPE (e.g., 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-X] (DSPE X) or N-[carbonyl-2′,3′-bis(methoxypolyethyleneglycol X)]-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-2arm PEGX)), PEGylated phosphoethanolamine (PE) (e.g., 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-X] (18:1 PEGX PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-X] (18:0 PEGX PE), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-X] (14:0 PEGX PE), or 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-5000] (16:0 PEGX PE)), PEGylated DPPE (e.g., N-(carbonyl-methoxypolyethyleneglycol X)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), PEGylated DMPE (e.g., N-(carbonyl-methoxypolyethyleneglycol X)-1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), PEGylated DPG (e.g., 1,2-dipalmitoyl-sn-glycerol, methoxypolyethylene glycol), PEGylated DSG (e.g., 1,2-distearoyl-sn-glycerol, methoxypolyethylene glycol), PEGylated DOG (e.g., 1,2-dioleoyl-sn-glycerol, methoxypolyethylene glycol), or PEGylated DMG (e.g., 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol), where X indicates an approximate weight average molecular weight (Mw) or approximate number average molecular weight (Mn), and where can be X 500, 3000, 2000, 1000, 750, 550, or 350.
- Exemplary sterols include, e.g., cholesterol, a derivative thereof, or any described herein. Exemplary zwitterionic lipids include DOPC, DPPC, DOPE, DPPE, POPC, DLPC, DSPC, DMPC, SOPC, or any described herein.
- Exemplary cationic lipids include 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-stearoyl-3-trimethylammonium-propane (18:0 TAP), 1,2-dipalmitoyl-3-trimethylammonium-propane (16:0 TAP), 1,2-dimyristoyl-3-trimethylammonium-propane (14:0 TAP), 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-amino-propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide (MVL5), ethylphosphocholine (ethyl PC) (e.g., 1,2-dimyristoleoyl-sn-glycero-3-ethylphosphocholine, 1-palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine, 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine, 1,2-distearoyl-sn-glycero-3-ethylphosphocholine, 1,2-dipalmitoyl-sn-glycero-3-ethylphosphocholine, 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine, or 1,2-dilauroyl-sn-glycero-3-ethylphosphocholine), dimethyldioctadecylammonium (DDAB), 1,2-dipalmitoyl-sn-glycero-O-ethyl-3-phosphocholine (EDPPC), or any described herein.
- The lipid layer of the particle can be composed of lipids and components in an amount similar to that provided by the lipid formulation. For instance, an exemplary lipid formulation comprising about 47 mol. % of a cationic lipid can provide a lipid layer (for a lipid-coated particle) that comprises 47 mol. % of that cationic lipid. Thus, any composition provided for a lipid formulation herein also provides a composition for the lipid layer.
- Targeting Ligands
- The lipid-coated particle can include one or more cell targeting species, cell penetrating peptides, fusogenic peptides, and/or targeting peptides. Such species can be included within the cargo, configured to be expressed by a plasmid of the cargo, located within the lipid layer, and/or provided by an external surface of the lipid layer (e.g., provided by the outer lipid layer). The composition of the lipid layer can include one or more components that facilitate ligand orientation, maximize cellular interaction, provide lipid stability, and/or confer enhanced cellular entry.
- In some instances, the targeting ligand can be a cell penetration peptide, a fusogenic peptide, or an endosomolytic peptide, which are peptides that aid a particle in translocating across a lipid bilayer, such as a cellular membrane or endosome lipid bilayer of the host cell. In one embodiment, the targeting ligand is optionally crosslinked onto a lipid layer surface of the outer lipid layer.
- Endosomolytic peptides are a sub-species of fusogenic peptides as described herein. Representative and preferred electrostatic cell penetration (fusogenic) peptides include an 8 mer polyarginine (NH2-RRRRRRRR-COOH, SEQ ID NO:1), among others known in the art, which are included in or on particles in order to enhance the penetration of into cells. Representative endosomolytic fusogenic peptides (“endosomolytic peptides”) include HSWYG peptide (NH2-GLFHAIAHFIHGGWHGLIHGWYGGC-COOH, SEQ ID NO:2), RALA peptide (NH2-WEARLARALARALARHLARALARALRAGEA-COOH, SEQ ID NO:3), KALA peptide (NH2-WEAKLAKALAKALAKHLAKALAKALKAGEA-COOH), SEQ ID NO:4), GALA (NH2-WEAALAEALAEALAEHLAEALAEALEALAA-COOH, SEQ ID NO:5) and INF7 (NH2-GLFEAIEGFIENGWEGMIDGWYG-COOH, SEQ ID NO:6), or fragments thereof, among others. In one instance, the targeting ligand includes an amino acid sequence having at least 80% sequence identity (e.g., at least 85%, 90%, 95%, or 99% sequence identity) to any one of SEQ ID NOs:1-6, or a fragment thereof.
- Proteins gain entry into the nucleus through the nuclear envelope. Yet other ligands can include a nuclear localization sequence (NLS), e.g., NH2-GNQSSNFGPMKGGNFGGRSSGPY GGGGQYFAKPRNQGGYGGC-COOH (SEQ ID NO:9), RRMKWKK (SEQ ID NO:10), PKKKRKV (SEQ ID NO:11), and KR[PAATKKAGQA]KKKK (SEQ ID NO:12), the NLS of nucleoplasmin, a prototypical bipartite signal comprising two clusters of basic amino acids, separated by a spacer of about 10 amino acids. Numerous other nuclear localization sequences are well known in the art. See, for example, LaCasse E C et al., “Nuclear localization signals overlap DNA- or RNA-binding domains in nucleic acid-binding proteins,” Nucl. Acids Res. 1995; 23:1647-56; Weis, K, “Importins and exportins: how to get in and out of the nucleus,” [published erratum appears in Trends Biochem. Sci. 1998 July; 23(7):23.5] Trends Biochem. Sci. 1998; 23:185-9; and Cokol M et al., EMBO Rep. 2000 Nov. 15; 1(5): 411-5, each of which is incorporated herein by reference in its entirety.
- Preferred ligands which may be used to target cells include peptides, affibodies, and antibodies (including monoclonal and/or polyclonal antibodies). In certain embodiments, targeting ligands selected from the group consisting of Fcγ from human IgG (which binds to Fcγ receptors on macrophages and dendritic cells), human complement C3 (which binds to CR1 on macrophages and dendritic cells), ephrin B2 (which binds to EphB4 receptors on alveolar type II epithelial cells), SP94 peptide (which binds to unknown receptor(s) on hepatocyte-derived cells), and MET receptor binding peptide. Exemplary, non-limiting SP94 peptides include SP94 free peptide (H2N-SFSIILTPILPL-COOH, SEQ ID NO:13), a SP94 peptide modified with C-terminal Cys for conjugation (H2N-SFSIILTPILPLGGC-COOH, SEQ ID NO:14), and a further modified SP94 peptide (H2N-SFSIILTPILPLEEEGGC-COOH, SEQ ID NO:15). Exemplary MET binding peptides include ASVHFPP (SEQ ID NO:16), TATFWFQ (SEQ ID NO:17), TSPVALL (SEQ ID NO:18), IPLKVHP (SEQ ID NO:19), and WPRLTNM (SEQ ID NO:20).
- Other exemplary targeting ligands include poly-L-arginine, including (R)n, where 6<n<12, such as an R12 peptide (e.g., RRRRRRRRRRRR (SEQ ID NO:21)) or an R9 peptide (e.g., RRRRRRRRR (SEQ ID NO:22)); a poly-histidine-lysine, such as a (KH)9 (e.g., KHKHKHKHKHKHKHKHKH (SEQ ID NO:23)); a Tat protein or derivatives and fragments thereof, such as RKKRRQRRR (SEQ ID NO:24), GRKKRRQRRRPQ (SEQ ID NO:25), GRKKRRQRRR (SEQ ID NO:26), GRKKRRQRRRPPQ (SEQ ID NO:27), YGRKKRRQRRR (SEQ ID NO:28), and RKKRRQRRRRKKRRQRRR (SEQ ID NO:29); a Cady protein or derivatives and fragments thereof, such as Ac-GLWRALWRLLRSLWRLLWRA-cysteamide (SEQ ID NO:30); a penetratin protein or derivatives and fragments thereof, such as RQIKIWFQNRRMKWKKGG (SEQ ID NO:31), RQIRIWFQNRRMRWRR (SEQ ID NO:32), and RQIKIWFQNRRMKWKK (SEQ ID NO:33); an antitrypsin protein or derivatives and fragments thereof, such as CSIPPEVKFNKPFVYLI (SEQ ID NO:34); a temporin protein or derivatives and fragments thereof, such as FVQWFSKFLGRIL-NH2 (SEQ ID NO:35); a MAP protein or derivatives and fragments thereof, such as KLALKLALKALKAALKLA (SEQ ID NO:36); a RW protein or derivatives and fragments thereof, such as RRWWRRWRR (SEQ ID NO:37); a pVEC protein or derivatives and fragments thereof, such as LLIILRRRIRKQAHAHSK (SEQ ID NO:38); a transportan protein or derivatives and fragments thereof, such as GWTLNSAGYLLGKIN LKALAALAKKIL (SEQ ID NO:39); a MPG protein or derivatives and fragments thereof, such as GALFLGFLGAAGSTMGAWSQPKKKRKV (SEQ ID NO:40); a Pep protein or derivatives and fragments thereof, such as KETWWETWWTEWSQPKKKRKV (SEQ ID NO:41), Ac-KETWWETWWTEWSQPKKKRKV-cysteamine (SEQ ID NO:42), and WKLFKKILKVL-amide (SEQ ID NO:43); a Bp100 protein or derivatives and fragments thereof, such as KKLFKKILKYL (SEQ ID NO:44) and KKLFKKILKYL-amide (SEQ ID NO:45); a maurocalcine protein or derivatives and fragments thereof, such as GDC(acm)LPHLKLC (SEQ ID NO:46); a calcitonin protein or derivatives and fragments thereof, such as LGTYTQDFNKFHTFPQTAIGVGAP (SEQ ID NO:47); a neurturin protein or derivatives and fragments thereof, such as GAAEAAARVYDLGLRRLRQRRRLRRERVRA (SEQ ID NO:48); and a human P1 protein or derivatives and fragments thereof, such as MGLGLHLLVLAAALQGAWSQPKKKRKV (SEQ ID NO:49).
- In one instance, the targeting ligand includes an amino acid sequence having at least 80% sequence identity (e.g., at least 85%, 90%, 95%, or 99% sequence identity) to any one of SEQ ID NOs:10-12 and 21-49 or a fragment thereof (e.g., having a length of about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, or more amino acids).
- Particle Characteristics and Surface Properties
- The lipid-coated particle can be characterized by any useful characteristic (e.g., overall charge, dimension, dispersity, etc.). In some embodiments, one or more optional targeting ligands can be present in or on a lipid layer. The particle can have any useful dimension, such as diameter, circumference, length, width, height, etc. Exemplary values for dimensions include, without limitation, greater than about 10 nm (e.g., greater than about 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 125 nm, 150 nm, 200 nm, 300 nm, 500 nm, 750 nm, 1 μm, 2 μm, 5 μm, 10 μm, 20 μm, or more) or of from about 2 nm to 500 nm (e.g., from 2 nm to 50 nm, 2 nm to 100 nm, 2 nm to 150 nm, 2 nm to 200 nm, 2 nm to 300 nm, 2 nm to 400 nm, 10 nm to 50 nm, 10 nm to 100 nm, 10 nm to 150 nm, 10 nm to 200 nm, 10 nm to 300 nm, 10 nm to 400 nm, 10 nm to 500 nm, 20 nm to 50 nm, 20 nm to 100 nm, 20 nm to 150 nm, 20 nm to 200 nm, 20 nm to 300 nm, 20 nm to 400 nm, 20 nm to 500 nm, 50 nm to 100 nm, 50 nm to 150 nm, 50 nm to 200 nm, 50 nm to 300 nm, 50 nm to 400 nm, 50 nm to 500 nm, 100 nm to 150 nm, 100 nm to 200 nm, 100 nm to 300 nm, 100 nm to 400 nm, 100 nm to 500 nm, 150 nm to 200 nm, 150 nm to 300 nm, 150 nm to 400 nm, 150 nm to 500 nm, 200 nm to 300 nm, 200 nm to 400 nm, or 200 nm to 500 nm).
- In particular embodiments, a plurality of particles is monodisperse, such as by having a polydispersity index (PdI) that is less than about 0.2 or by having a PdI that is of from about 0.05 to about 0.2 (e.g., from 0.05 to 0.1, 0.05 to 0.15, 0.1 to 0.15, 0.1 to 0.2, or 0.15 to 0.2). In some embodiments, the monodisperse particles range in a size of from about 20 nm to about 300 nm (e.g., from 50 nm (+/−10 nm) to 150 nm (+/−15 nm)). In other embodiments, the particle (or a plurality of particles) has a charge (or a net charge) that is near neutral (e.g., a zeta potential of from about +5 mV to −5 mV).
- In certain alternative embodiments, the present invention is directed to particles of a particular size (diameter) ranging from about 0.5 to about 30 nm, about 1 nm to about 30 nm, often about 5 nm to about 25 nm (preferably, less than about 25 nm), often about 10 to about 20 nm, for administration via intravenous, intramuscular, intraperitoneal, retro-orbital, and subcutaneous injection routes. These particles can be monodisperse and provide colloidally stable compositions.
- The surface properties of the particle can be optimized in any useful manner. For instance, the lipid layer can have an appropriate charge (e.g., approximately net neutral charge), can include appropriate targeting ligands to promote their cell-specific binding and internalization, and can include useful ligand (e.g., to promote endosomal escape or nuclear localization within host cells).
- Any useful ligand can be employed. The type and density of targeting ligands can be optimized to enhance uptake by the target. Exemplary ligands include a peptide that binds to ephrin B2, which we identified using phage display, to target Vero cells; Fcγ to target THP-1 cells and primary alveolar macrophages; the ‘GE11’ peptide (see, e.g., Li Z et al., FASEB J2005; 19: 1978-85) to target A549 cells and primary alveolar epithelial cells; the ‘SP94’ peptide (see, e.g., Lo A et al., Molec. Cancer Therap. 2008; 7:579-89) to target HepG2 cells and primary hepatocytes; human complement C3, which binds to receptors on macrophages and dendritic cells; or the ‘H5WYG’ peptide, which ruptures the membranes of acidic intracellular vesicles via the ‘proton sponge’ mechanism (see, e.g., Moore N M et al., J. Gene. Med. 2008 10: 1134-49).
- Other ligands include a peptide (e.g., a peptide zip code or a cell penetrating peptide), an endosomolytic peptide, an antibody (including fragments thereof), affibodies, a carbohydrate, an aptamer, a cluster of differentiation (CD) protein, or a self-associated molecular pattern (SAMP) (e.g., as described in Lambris J D et al., Nat. Rev. Microbiol. 2008; 6(2):132; and Poon I K H, Cell Death Differ. 2010; 17:381-97, each of which is incorporated herein by reference in its entirety). Exemplary CD proteins include CD47 (OMIM Entry No. 601028, a marker of self that allows RBC to avoid phagocytosis), CD59 (OMIM Entry No. 107271, a marker that prevents lysis by complement), C1 inhibitor (C1INH, OMIM Entry No. 606860, a marker that suppresses activation of the host's complement system), CD200 (OMIM Entry No. 155970, an immunosuppressive factor), CD55 (OMIM Entry No. 125240, a marker that inhibits the complement cascade), CD46 (OMIM Entry No. 120920, a marker that inhibits the complement cascade), and CD31 (OMIM Entry No. 173445, an adhesion regulator and a negative regulator of platelet-collagen interactions). Each recited OMIM Entry is incorporated herein by reference in its entirety.
- Any other useful ligand can be employed, such as those identified by the ‘BRASIL’ (Biopanning and Rapid Analysis of Selective Interactive Ligands) method (see, e.g., Giordano R J et al., Nat. Med. 2001; 7:1249-53; Giordano R J et al., Proc. Natl Acad. Sci. USA 2010; 107(11):5112-7; and Kolonin M G et al., Cancer Res. 2006; 66:34-40) to identify novel targeting peptides and single-chain variable fragments (scFvs) via phage display (see, e.g., Giordano R J et al., Chem. Biol. 2005; 12:1075-83; Giordano R J et al., Proc. Natl Acad. Sci. USA 2010; 107(11):5112-7; Kolonin M G et al., Cancer Res. 2006; 66:34-40; Tonelli R R et al., PLoS Negl. Dis. 2010; 4:e864; Lionakis M S et al., Infect. Immun. 2005; 73:7747-58; and Barbu E M et al., PLoS Pathog. 2010; 6:e1000726).
- Methods
- The lipid-coated particles herein can be employed in any useful manner. The present particles can be adapted to recognize the target and, if needed, deliver the one or more cargos to treat that target. Exemplary targets include a cell, a pathogen, an organ (e.g., dermis, vasculature, lymphoid tissue, liver, lung, spleen, kidneys, heart, brain, bone, muscle, etc.), a cellular target (e.g., targets of the subject, such as a human subject, including host tissue, host cytoplasm, host nucleus, etc., in any useful cell, such as e.g., hepatocytes, alveolar epithelial cells, and innate immune cells, etc.); as well as targets for exogenous cells and organisms, such as extracellular and/or intracellular components of a pathogen, e.g., bacteria), a molecular target (e.g., within the subject or the exogenous cell/organism, such as pathogen DNA, host DNA, pathogen RNA, pathogen proteins, surface proteins or carbohydrates of any subject or exogenous cell), etc.
- In one instance, the particle is employed to target a host (e.g., a subject), a pathogen, or both (e.g., thereby treating the subject and/or the target). Exemplary pathogens include a bacterium, such as Bacillus (e.g., B. anthracis), Enterobacteriaceae (e.g., Salmonella, Escherichia coli, Yersinia pestis, Klebsiella, and Shigella), Yersinia (e.g., Y. pestis or Y. enterocolitica), Staphylococcus (e.g., S. aureus), Streptococcus, Gonorrheae, Enterococcus (e.g., E. faecalis), Listeria (e.g., L. monocytogenes), Brucella (e.g., B. abortus, B. melitensis, or B. suis), Vibrio (e.g., V. cholerae), Corynebacterium diphtheria, Pseudomonas (e.g., P. pseudomallei or P. aeruginosa), Burkholderia (e.g., B. mallei or B. pseudomallei), Shigella (e.g., S. dysenteriae), Rickettsia (e.g., R. rickettsii, R. prowazekii, or R. typhi), Francisella tularensis, Chlamydia psittaci, Coxiella burnetii , Mycoplasma (e.g., M. mycoides), etc.; mycotoxins, mold spores, or bacterial spores such as Clostridium botulinum and C. perfringens; a virus, including DNA or RNA viruses, such as Adenoviridae (e.g., adenovirus), Arenaviridae (e.g., Machupo virus), Bunyaviridae (e.g., Hantavirus or Rift Valley fever virus), Coronaviridae, Orthomyxoviridae (e.g., influenza viruses), Filoviridae (e.g., Ebola virus and Marburg virus), Flaviviridae (e.g., Japanese encephalitis virus, hepatitis C virus, and Yellow fever virus), Hepadnaviridae (e.g., hepatitis B virus), Herpesviridae (e.g., herpes simplex viruses, herpesvirus, cytomegalovirus, Epstein-Barr virus, or varicella zoster viruses), Papillomaviridae (e.g., papilloma viruses), Papovaviridae (e.g., papilloma viruses), Paramyxoviridae (e.g., respiratory syncytial virus, measles virus, mumps virus, or parainfluenza virus), Parvoviridae, Picornaviridae (e.g., polioviruses and hepatitis A virus), Polyomaviridae, Poxviridae (e.g., variola viruses or vaccinia virus), Reoviridae (e.g., rotaviruses), Retroviridae (e.g., human T cell lymphotropic viruses (HTLV) and human immunodeficiency viruses (HIV)), Rhabdoviridae (e.g., rabies virus), and Togaviridae (e.g., encephalitis viruses, yellow fever virus, and rubella virus)); a protozoon, such as Cryptosporidium parvum, Encephalitozoa, Plasmodium, Toxoplasma gondii, Acanthamoeba, Entamoeba histolytica, Giardia lamblia, Trichomonas vaginalis, Leishmania, or Trypanosoma (e.g., T. brucei and T. cruzi); a helminth, such as cestodes (tapeworms), trematodes (flukes), or nematodes (roundworms, e.g., Ascaris lumbricoides, Trichuris trichiura, Necator americanus, or Ancylostoma duodenale); a parasite (e.g., any protozoa or helminths described herein); or a fungus, such as Aspergilli, Candidae, Coccidioides immitis, and Cryptococci. Other pathogens include a multi-drug resistant (MDR) pathogen, such as MDR forms of any pathogen described herein. Additional pathogens are described in Cello J et al., Science 2002; 297:1016-8; Gibson D G et al., Science 2010; 329:52-6; Jackson R J et al., J. Virol. 2001; 75:1205-10; Russell C A et al., Science 2012; 336:1541-7; Tumpey T M et al., Science 2005; 310:77-80; and Weber N D et al., Virology 2014; 454-455:353-61, each of which is incorporated herein by reference in its entirety.
- The present invention can be employed to treat an infection (e.g., a viral infection). Exemplary infections include an encephalitis infection, a viral infection, a bacterial infection, etc. Infections can arise from a virus, such as a mosquito-borne viral pathogen, an encephalitis virus (e.g., Venezuelan equine encephalitis virus (VEEV)), herpes virus (e.g., herpes simplex virus, varicella-zoster virus, and Epstein-Barr virus), rabies virus, poliovirus, measles virus, an arbovirus (e.g., St. Louis encephalitis virus and West Nile encephalitis virus), bunyavirus (e.g., La Crosse strain, California encephalitis virus, etc.), arenavirus (e.g., lymphocytic choriomeningitis virus), reovirus (e.g., Colorado tick virus), henipavirus, flavivirus (e.g., Japanese encephalitis virus (JEV), St. Louis encephalitis virus, etc.), enterovirus, and Powassan virus. Alternatively, the infection can arise from bacteria, fungi, and/or protozoa.
- Compositions and Formulations
- The present lipid-coated particles can be formulated in any useful manner. For instance, the formulation can be optimized for subcutaneous (SC), intranasal (IN), aerosol, intravenous (IV), intramuscular (IM), intraperitoneal (IP), oral, topical, transdermal, or retro-orbital delivery. Any useful dosages can be employed within the formulations. Exemplary dosages include, e.g., 200 mg/kg. The formulation or composition can include a plurality of particles (e.g., an effective amount thereof) and an optional pharmaceutically acceptable excipient (e.g., any described herein).
- In some instances, the pharmaceutical composition includes a population of particles (e.g., any described herein) in an amount effective for modulating or modifying a target gene within a subject in combination with a pharmaceutically acceptable carrier, additive, or excipient. In other instances, the composition further includes a drug, a therapeutic agent, etc., which is not disposed as cargo within the particle.
- The composition can be formulated in any useful manner with a plurality of particles. Such formulations can be included with any useful medium, excipient (e.g., lactose, saccharide, carbohydrate, mannitol, leucine, PEG, trehalose, etc.), additive, propellant, solution (e.g., aqueous solution, such as a buffer), additive, preservative, carrier (e.g., aqueous saline, aqueous dextrose, glycerol, or ethanol), binder (e.g., saccharide, cellulose preparation, starch paste, or methyl cellulose), filler, or disintegrator.
- Pharmaceutical compositions according to the present invention include an effective population of lipid-coated particles herein formulated to affect an intended result (e.g., immunogenic result, therapeutic result and/or diagnostic analysis, including the monitoring of therapy) formulated in combination with a pharmaceutically acceptable carrier, additive or excipient. The particles within the population of the composition may be the same or different depending upon the desired result to be obtained. Pharmaceutical compositions according to the present invention may also comprise an addition bioactive agent or drug, such as an antibiotic or antiviral agent.
- Formulations and compositions containing the particles according to the present invention may take the form of liquid, solid, semi-solid or lyophilized powder forms, such as, for example, solutions, suspensions, emulsions, sustained-release formulations, tablets, capsules, powders, suppositories, creams, ointments, lotions, aerosols, patches or the like, preferably in unit dosage forms suitable for simple administration of precise dosages.
- Methods for preparing such dosage forms are known or apparent to those skilled in the art; for example, see Remington's Pharmaceutical Sciences (17th Ed., Mack Pub. Co., 1985). The composition to be administered will contain a quantity of the selected compound in a pharmaceutically effective amount for therapeutic use in a biological system, including a patient or subject according to the present invention.
- New World alphaviruses affect North, South, and Central America and pose a major public health threat as they are highly infectious and can result in fatal encephalitis in humans [1-3]. One of these alphaviruses, the Venezuelan Equine Encephalitis Virus (VEEV), is classified as a Category B Agent by the CDC and NIAID due to its amenability to aerosolization while remaining highly infectious and the lack of controlled vaccines and antivirals against the virus [3]. Because of its potentially debilitating health consequences, low infectious dose in humans, and stability in storage, VEEV is a potential bioterrorism agent and has been previously stockpiled in the US and USSR [2, 3].
- In addition to its use as a bioterrorism agent, natural VEEV outbreaks result in equine and human infections in North and South America, causing high rates of fatality in equines (85%) and chronic neurological complications in humans [3-5]. Infected humans experience influenza-like symptoms, and 14% of infections result in neurological complications and sequelae, including disorientation, ataxia, depression, and convulsions [2, 5]. In one percent of cases, human infections result in mortality [4, 5]. Thus, developing strategies to inhibit VEEV infection is critical to minimizing fatalities and complications in cases of bioterrorism and natural outbreaks. For all these reasons, VEEV poses a major public health risk due to its amenability to use as a bioterrorism agent and its severe health consequences in humans and equines.
- Several small molecule drugs have been developed that inhibit VEEV, but many are limited by high toxicity or low efficacy [6-11]. Recently, a highly effective small molecule inhibitor of VEEV was developed with the assistance of a high throughput, cell-based screen [4, 6]. Referred to as ML336, this molecule was found to have a EC90 of 170 nM against a VEEV vaccine strain (TC-83) and reduce viral titer by 630,000-fold at nanomolar concentrations. In addition, intraperitoneal administration of ML336 to mice infected with TC-83 resulted in a 71% survival rate as compared to the 14% survival rate observed in untreated mice. While the potency of this drug at nanomolar concentrations and in in vivo studies is encouraging, ML336 has limited solubility (0.04 mg/mL in PBS, pH 7.4) and limited stability (reduction of 17% and 35% of drug in PBS and mouse plasma, respectively, after 3 hours) in aqueous solutions [4], potentially reducing its effectiveness.
- Thus, ML336 is a recently developed small molecule inhibitor of VEEV, shown to effectively reduce VEEV strain TC-83 both in vitro and in vivo, but its limited solubility and stability could hinder its use in future applications. To improve drug solubility and stability, we investigated utilizing a nanoparticle based platform to deliver ML336 for VEEV inhibition both in vitro and in vivo. In particular, lipid-coated mesoporous silica nanoparticles (LC-MSNs) were employed. The large surface area of the MSN core promotes hydrophobic drug loading, while the liposome coating enables enhanced circulation time and biocompatibility, thereby providing a platform for ML336 delivery.
- Mesoporous silica nanoparticles (MSNs) have been used in drug delivery systems to improve drug stability and solubility, protect cargo, target specific tissues, and enhance drug circulation time and controlled release [12, 13]. MSNs have a narrow size distribution and can be optimized for various drug delivery applications by tuning particle size, pore size, surface properties, and the porous structure [14, 15]. Established methods enable formation of MSNs with uniform and tunable pore size, endowing MSNs with a large and uniform surface area for drug adsorption (600-1000 m2/g) [12-15]. This property is particularly advantageous for loading water insoluble or unstable drugs, as the large surface area acts as a reservoir for hydrophobic drug in aqueous solution and can improve drug efficacy in vivo [16, 17]. In addition, MSNs are stable in non-aqueous solutions and permit loading of hydrophobic drugs in organic solvents, giving them a distinct advantage over polymeric or liposomal nanoparticle delivery systems [18].
- While MSNs are a promising carrier for ML336, drug-loaded MSNs can have low colloidal stability and are subject to aggregation in physiological solutions, reducing circulation time and preventing desirable cell uptake [19, 20]. In addition, premature release of cargo from MSNs can be problematic [21]. In order to overcome these challenges, we investigated the application of a lipid-based coating to the exterior of ML336-loaded MSNs.
- MSNs coated with supported lipid bilayers (lipid-coated MSNs (LC-MSNs)) have been employed in drug and protein delivery applications to improve colloidal stability and subsequent circulation time, biocompatibility, cargo loading and release, and tissue-specific targeting [19, 21-23]. The application of a supported lipid bilayer to the exterior of the MSN (essentially encapsulating the MSN with a liposome) can improve colloidal stability in physiological solutions [19, 20] and prevent cargo release prior to cell internalization or some other external trigger [21]. In addition, a lipid bilayer coating offers an additional surface that can be functionalized independently of the MSN surface for tissue-specific targeting [19, 21, 22, 24]. Finally, the inherent instability and broad size distribution of liposomes can be overcome when combined with MSNs to form LC-MSNs [21, 22, 25]. Thus, LC-MSNs harness the advantages and overcome the obstacles associated with MSNs and liposomes in one versatile platform for small molecule delivery.
- Here, we highlight the use of LC-MSNs for ML336 delivery to inhibit VEEV (see, e.g.,
FIG. 2 ). LC-MSN characterization revealed uniformly sized particles coated with a lipid bilayer and good colloidal stability, as assessed by dynamic light scatter analysis, zeta potential measurements, and cryogenic electron microscopy. The delivery vehicle was able to load and release ML336 in a manner that inhibited virus in vitro. In particular, LC-MSNs were found to load 20±3.4 μg ML336/mg LC-MSN and to release 6.6±1.3 μg/mg over the course of 24 hours. ML336-loaded LC-MSNs inhibited VEEV in vitro in a dose-dependent manner and by about 4-6 orders of magnitude as compared to untreated controls. In addition, in vitro studies suggested that additional release of ML336 occurs after cellular internalization, in which studies suggest that this was mediated through a clathrin-mediated endocytosis pathway. - Finally, ML336 loaded LC-MSNs showed viral inhibition in an in vivo murine model of VEEV infection. In vivo safety studies in C3H/HeN mice shows that LC-MSNs were not toxic when dosed at 0.11 mg LC-MSNs/kg daily for four days. In addition, ML336-loaded LC-MSNs showed significant reduction in brain viral titer in VEEV TC-83 infected mice as compared to PBS treated controls. Overall, to our knowledge, this work demonstrates the first use of a nanoparticle-based system for the delivery of ML336. The successful inhibition of virus achieved with this platform could have widespread benefit in combatting VEEV and other viral infections resulting from bioterrorism or natural causes. Additional details follow.
- The following provide exemplary materials and methods employed for data described herein.
- MSN fabrication and characterization: Both small and large batch syntheses of monosized hexagonally-structured MSNs were prepared as previously described [19,41,42] with modifications. MSNs (up-scaled batch of hexagonal small pore particles) were synthesized in a large batch format by dissolving 1.45 g of cetyl trimethylammonium bromide (CTAB) (Sigma) in 750 mL of a 0.32 M aqueous ammonium hydroxide solution in a parafilm covered beaker (1 L). The beaker was placed in a 50° C. silicon oil bath for 2 hr (hours) with continuously stirring at high speed (650 rpm). A tetraethyl orthosilicate (TEOS) solution, prepared at 0.88M by combining 3 mL of TEOS (Sigma) with 12 mL of 100% ethanol, was subsequently added to the CTAB surfactant solution. The reaction was stirred vigorously for 1 hr uncovered and then incubated overnight (˜18 hrs) in a 50° C. silicone oil bath without stirring. The remaining volume was transferred to a 500 mL glass bottle for an overnight hydrothermal treatment at 70° C. The MSN solution was aliquoted into eight 50 mL tubes and centrifuged at 50K×g for 15 minutes (min). The pellets were resuspended, washed twice with 100% ethanol, and combined into 4 tubes for a second wash. After washing, CTAB removal was achieved by resuspending particles in 100 mL of 6 g/L ammonium nitrate in ethanol. Tubes were placed in a 60° C. bath with sonication for 1.5 hr or at 40° C. with sonication for 30 min. Particles were collected by centrifugation, washed with ethanol (with 90% ethanol and then 100% ethanol; or with 95% ethanol), collected by centrifugation, resuspended in 100 mL of a 1% HCl in ethanol solution, and sonicated for 1.5 hr at 60° C. or twice for 30 min at 40° C. Particles were once again collected by centrifugation, washed with 90% ethanol and then by 100% ethanol, collected by centrifugation and resuspended in 40 mL of 100% ethanol. The MSN suspension was passed through a 1 μm filter to remove large aggregates and weighed after particle desiccation. Size and zeta potential were measured using a Zetasizer instrument (Malvern).
- Fluorescently labeled (Cy3) nanoparticles were synthesized in a small scale format by dissolving 250 mg of CTAB in 150 mL of 0.32 M ammonium hydroxide solution in a 250 ml beaker. The reaction was covered with parafilm and heated to 50° C. in a silicone oil bath for 1 hr with continuously stirring at high speed (650 rpm). A Cy3 dye solution was prepared by dissolving 3 mg of Cy3-NHS (ThermoFisher) in 1 ml of 100% ethanol using sonication, followed by the addition of 2.5 μL of APTES (Sigma). The Cy3 solution was allowed to incubate at room temperature without light for 1 hr. A TEOS solution composed of 3 ml of 0.88 M TEOS, was combined with the Cy3-APTES solution, and then added to the CTAB solution and stirred uncovered at 50° C. After 1 hr, the solution was left overnight (˜18 hr) in the 50° C. oil bath without stirring. The remaining volume was transferred to a 100 mL glass bottle for an overnight hydrothermal treatment at 70° C. All centrifugation steps were carried out at 50,000 x g for 15 min. The MSN solution was collected by centrifugation and particles were washed twice with 100% ethanol.
- CTAB removal was carried out by resuspending particles in 20 mL of a 6 g/L ammonium nitrite in ethanol, and sonicated at 40° C. for 20 min. Particles were collected by centrifugation, washed with 95% ethanol, collected by centrifugation, resuspended in 20 mL of an ethanolic HCl solution (1%) and sonicated twice for 20 min at 40° C. Cy3-labelled particles were washed with 90% ethanol followed by 100% ethanol, collected by centrifugation and resuspended in 12 mL of 100% ethanol.
- Prior to use, all MSN suspensions were passed through a 1 μm filter to remove any eventual large aggregates. Quantification was carried out by weight after desiccation of three 500 μL aliquots. Size and zeta potential were measured using a Zetasizer instrument (Malvern Instruments, Ltd). Morphology was assessed with TEM (JEOL 2010). For porosimetry measurements nitrogen sorption data was collected at 77 K with a Quantachrome AutoSorb-iQ2 sorption analyzer, after degassing the samples under vacuum at 333 K. Surface areas were determined using the Brunauer-Emmett-Teller (BET) model. Non-Local Density Functional Theory (NLDFT) was used to calculate pore size distributions and surface areas assuming the surface to be silica with cylindrical pores. Pore size distributions were also calculated according to the Barrett Joyner Halenda (BJH) method. The SEM analysis was carried out using a probe-corrected Hitachi HF5000 TEM/STEM at 200 kV in STEM mode. The STEM unit is equipped with a secondary electron (SE) detector in addition to the annular dark field (ADF), annular bright field (ABF) and bright field (BF) detectors, which allow simultaneous secondary and transmitted election imaging to obtain information from the surface (SE) and bulk (ADF and BF) of the nanomaterial.
- LC-MSN fabrication and characterization: Liposomes were prepared by combining 77.5
mol % 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 20 mol % cholesterol, and 2.5mol % 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000) (Avanti Polar Lipids) in chloroform at a 5 mg/mL concentration. Lipid films were prepared using a rotary evaporator (Buchi Corp.), incubated under vacuum overnight, and rehydrated at a 5 mg/mL concentration in a 0.5×PBS (Gibco/Life Technologies), 4 mM MgCl2 solution for 30 minutes at 55° C. The lipid solution was purged with nitrogen for two minutes and then dispersed with an ultrasonication probe (Branson Sonifier, Emerson US). Lipids were sonicated under nitrogen at 10-12 watts for 4 min, followed by a 2 min rest period, repeated twice. Lipids were centrifuged at 16,000 RCF for 20 min to remove any debris deposited into the lipid solution from the sonication probe. - Loaded LC-MSNs were prepared by resuspending 1 mg MSNs in 10 μL water followed by overnight incubation at 4° C. in 100 μl of 1 mg/mL ML336 (Caymen Chemicals) in dimethyl sulfoxide (DMSO) (100 μL DMSO for unloaded LC-MSN groups). To form LC-MSNs, the resulting liposomes were combined with MSNs under bath sonication while pipetting at a 5:1 mass ratio of liposomes:nanoparticles. Particles were then centrifuged at 21,000 RCF to remove liposomes that were not fused with MSNs. When used immediately, resulting LC-MSNs were rinsed twice by resuspending in 1 mL PBS, centrifuging at 21,000 RCF, and removing supernatant. For storing LC-MSNs, particles were rinsed once in PBS and then resuspended in a 9 wt % sucrose solution in PBS, flash frozen in liquid nitrogen, and stored at −80° C. Prior to use (e.g. cryo-EM analysis and all in vitro and in vivo studies), particles were thawed and rinsed once in PBS. For animal studies with viral infection, LC-MSNs were prepared in 2 mg aliquots, combined, and redistributed into 1 or 1.5 mg aliquots before freezing for storage.
- Dynamic light scattering (DLS) for particle hydrodynamic diameter and polydispersity index (PDI), and zeta potential measurements were obtained using a Malvern Zetasizer. For cryo-EM analysis, unloaded and loaded LC-MSNs were vitrified using Vitrobot (Thermo Fisher Scientific) as previously described [43]. Briefly, 3 μL of particles suspension was placed on a C-flat grid (Protochips, Inc.) with 2 μm diameter holes, blotted with filter paper, and plunged into liquid ethane for flash freezing. Frozen grids were stored under liquid nitrogen and were transferred to electron microscope JEM 2200FS (JEOL Ltd). Grids were imaged at 200 keV using DE-20 (Direct Detector Inc.) direct electron detector camera. The 2200FS microscope had Field Emission electron source and an omega-type electron energy filter to remove inelastically scattered electrons from image formation. The energy selecting slit was set to 20 eV. DE-20 camera was used in “movie” mode with frame rate of 25 frames/s. Off-line frame alignment was performed with DE_process_frames.py script provided by Direct Electron Inc. Images were collected at 40,000× indicated magnification, the pixel size on the specimen scale corresponded to 1.5 Å/pixel. Images were collected with 1.5 to 2.6 μm defocus range. Cryo-EM images were analyzed for lipid bilayer thickness using ImageJ. Fifty particles were analyzed from each of the ML336 loaded LC-MSN and unloaded LC-MSN groups.
- LC-MSN ML336 loading and release studies: The concentration of ML336 was determined by correlating sample (supernatants) spectroscopic absorption measurements at 320 nm (Nanodrop, ThermoFisher Scientific) with ML336 standard curves prepared in the 5 mg/mL liposome solution (described above) or PBS (
FIG. 4C ). Loading of ML336 in LCMSNs was calculated using the following formula: Total mass loaded=Initial mass of ML336 added −[(mass of ML336 in the supernatant after combination with the lipids)+(mass of ML336 in the supernatant of PBS wash 1)+(mass of ML336 in the supernatant of PBS wash 2)]. A dataset of six replicates highlights further how loading and release was calculatedFIG. 4D ). Briefly, to determine the mass of ML336 loaded, the mass of ML336 in the supernatant from each wash step (one after lipid application and two in PBS, labeled A, B, and C, respectively, inFIG. 4D ) was subtracted from the total mass of ML336 loaded, 100 μg (100 μg−(A+B+C);FIG. 4D ). The total loading was then reported on a per mass LC-MSN basis. - For release studies, rinsed LC-MSNs were resuspended in 1 mL of PBS and incubated at room temperature. Release of ML336 was measured by removing 100 μL aliquots from each sample, centrifuging the aliquot at 21,000 RCF, and measuring the concentration of ML336 in the supernatant at 0.25, 0.5, 0.75, 1, 2, 3, 4, 24, and 96 hr timepoints. Cumulative mass released was calculated by averaging the mass observed in each sample at each timepoint. Cumulative percent released was calculated by averaging the percent released for each sample at each timepoint using the following formula: Percent released=100%*(mass released at timepoint X/total mass loaded).
- LC-MSN internalization studies: All cells were maintained at 37° C. and 5% CO2. Cells were maintained in Dulbecco's Modified Eagle Medium (DMEM, Gibco/Life Technologies; HeLas) or Minimum Essential Medium a (aMEM, Gibco/Life Technologies; Veros) supplemented with 10 vol % FBS (Hyclone), 10,000 IU/mL penicillin, and 10,000 μg/ml streptomycin (MP Biomedicals).
- For endocytosis inhibitor studies with LC-MSNs, HeLa cells were plated overnight in complete medium on 12-well plates for image analysis or flow cytometry. Unless otherwise indicated, all inhibitors were purchased from EMD Millipore. The following inhibitors were initially resuspended in DMSO and were then diluted in complete medium to obtain the final concentrations indicated: the vacuolar H+-ATPase inhibitor bafilomycin A (BAF)(100 nM), the cationic amphiphilic compound chlorpromazine (CPZ; 6.5 μg/ml; Sigma),
dynamin 2 inhibitor dynasore (DYN) (80 μM), wortmannin (Wort) (150 ng/ml), p21-activated kinase inhibitor III (IPA-3) (10 μM), and phorbol 12-myristate 13-acetate (PMA) (10 μM). Cells were incubated with inhibitor treatments for lh prior and during incubation with Cy3 labeled LC-MSNs, or the pathogens Vesicular Stomatitis virus (VSV strain Indiana 1) and Rift Valley fever virus (RVFV strain MP-12) used as specificity controls for clathrin-mediated endocytosis and caveola-mediated endocytosis, respectively. - For image analysis studies, the cells were washed at 5 hr post-LC-MSN addition with PBS twice and then subjected to brightfield and fluorescent microscopy.
- For flow cytometry experiments, cells were washed with PBS twice at 16 hr post LC-MSN addition or virus infection, then prepared for analysis on a BD Accuri C6 instrument (Becton, Dickerson and Company). Flow cytometry data was analyzed by FCS Express v6 software (De Novo Software).
- For confocal microscopy imaging of cellular association with LC-MSNs, HeLa cells were seeded overnight onto No. 1.5 glass cover slips in 6-well plates at a density of 0.75×105 cells per well. Fluorescent LC-MSNs were then added at 25 μg MSN/well for 45 min or 20 hr. After incubation, cells were washed with PBS, fixed with 4% paraformaldehyde in PBS for 15 min with prewarmed solutions followed by overnight storage at 4° C., washed twice with PBS, and made permeable with 0.1% Triton-X in PBS for 15 min. Cells were then blocked with 1% BSA in PBS for 20 min and then labeled with 5 units/1 ml Alexa Fluor 647 phalloidin (ThermoFisher) and Alexa Fluor 488 anti-a-tubulin antibody (Invitrogen) in blocking buffer for 1 hr. After washing with PBS, slides were mounted using Prolong Gold with DAPI (ThermoFisher). Confocal images were acquired with a 63X/1.4NA oil objective in sequential scanning mode using a Leica TCS SP8 confocal microscope. Three-dimensional cell images were isosurface rendered using the Leica Application Suite Advanced Fluorescence 3D analysis software.
- In vitro viral inhibition: All cells were maintained at 37° C. and 5% CO2. Cells were maintained in Dulbecco's Modified Eagle Medium (DMEM, Gibco/Life Technologies; HeLas) or Minimum Essential Medium a (aMEM, Gibco/Life Technologies; Veros) supplemented with 10 vol % FBS (Hyclone), 10,000 IU/mL penicillin, and 10,000 μg/ml streptomycin (MP Biomedicals).
- The TC-83 virus was obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH (NR-63), and was propagated in Vero cells. Cells were infected at a multiplicities of infection (MOI) of 0.1 and cultured for two days. Supernatant was collected and the concentration of plaque forming units (PFUs) was determined using a standard plaque assay with an agarose overlay (1:1 2×Modified Eagle Medium (Gibco/Life Technologies; 8 vol % FBS, 10,000 IU/mL penicillin, and 10,000 m/ml streptomycin):1.5 wt % agarose (Invitrogen)). Cells were fixed and stained with an ethanol-based crystal violet solution (0.14 wt % crystal violet (Sigma-Aldrich), 21 vol % ethanol) and plaques were counted manually to determine PFU/mL.
- HeLa cells at 80-90% confluency in 12 well plates were pretreated with 25 μg LC-MSNs in 100 μL Opti-MEM Reduced Serum Media (Gibco/Life Technologies) for 1 hr, then infected with TC-83 at 0.1 MOI for 30-60 min. Virus was then removed, cells were rinsed three times in PBS, and treatments were added back for the remainder of the time course in 1 mL DMEM. Supernatants were taken at 24, 48, and 72 hrs. Phase images were taken at the same timepoints using an inverted microscope (Olympus-IX70).
- For experiments with pre-released LC-MSNs, LC-MSNs were incubated in Opti-MEM at a concentration of 2.5 μg/mL for 4 hrs at room temperature. LC-MSNs were then centrifuged at 20,000 RCF, and supernatant was collected. Particles were resuspended at 2.5 μg/mL in Opti-MEM and immediately added to cell cultures as described above. The supernatant of 25 μg of particles (100 μL) was also immediately added to cells.
- The concentration of PFUs in supernatants was determined using a standard plaque assay on VERO cells in 12 well plates as described above. Serial dilutions of supernatants were prepared in aMEM and used to infect cells for 30-60 min. Due to the minimum concentration of virions in supernatants required for detection in plaque assays, the limit of detection (indicated on each graph) was 100 PFU/mL.
- In vivo viral inhibition: All animal work was conducted in accordance with protocols approved by the Lawrence Livermore National Laboratory Institution Animal Care and Use Committee. For safety studies, five 6-8 week old C3H/HeN mice were injected intraperitoneal (IP) with 1 mg LC-MSNs in 200 μL PBS, and three mice were injected with 200 μL PBS only. Mice were monitored for 15 days and weighed on
days day 15, animals were euthanized; and lungs, livers, spleens, kidneys, and brains were dissected from three animals in each group. - An established C3H/HeN mice model of VEEV infection was used to assess the ability of ML336-loaded LC-MSNs to inhibit viral infection [8]. In the first animal study, animals were divided into four groups of ten animals each: ML336 loaded LC-MSNs, unloaded LC-MSNs, free drug, and PBS groups. For ML336 loaded and unloaded LC-MSNs groups, 1 mg of LC-MSNs was mixed with 200
μL 1 wt % carboxymethylcellulose sodium (Sigma-Aldrich) in PBS. For free drug groups, 20 μL of 1 mg/mL ML336 in DMSO was mixed into 200 μL of 1 wt % carboxymethylcellulose sodium in PBS, resulting in 20 μg ML336 per injection, similar to what is loaded in 1 mg ML336-loaded LC-MSNs. Mice were injected twice a day IP for 4 days. Four hours after the first injection onday 1, the mice were infected with an intranasal instillation of 50 μl of TC-83 containing a total of 108 PFU. - Mice were monitored for weight and clinical signs of disease each day post-infection and assigned a clinical score from 0-4 (mild to severe): 0=bright, alert, responsive and active, animals exhibit normal grooming and social behavior, no loss of appetite; 1=mild clinical signs of infection such as coat ruffling and loss of appetite; 2=pronounced decrease in activity and responsiveness to stimulation, ruffled coat and rapid, shallow breathing, obvious neurological impairment such as trouble ambulating and hunching; 3=moribund, eyes closed completely, labored breathing, no activity and unresponsive to tail tug, or animals that have lost >25% of their body weight, in which mice that met this criterion were euthanized; and 4=found dead.
- The second animal study was conducted as described above with the following changes. LC-MSN masses for both loaded and unloaded LC-MSN groups were increased to 1.5 mg. For free drug groups, 15 μL of 2 mg/mL ML336 in DMSO was mixed into 185 μL of 1 wt % carboxymethylcellulose in PBS, resulting in 30 μg ML336 per injection, similar to what is loaded in 1.5 mg ML336-loaded LC-MSNs. On
day 5, five mice from each group were euthanized and dissected for brain, spleen, kidney, liver, and serum. Organs were homogenized using disposable tissue grinders, and tissue lysate was assessed for viral load using a standard plaque assay as described above. - Statistical Analysis: All results are depicted as mean±standard deviation. Analysis of Variance (ANOVA) was used to identify significant factors and interactions, then Tukey's post hoc test (significance level p<0.05) was used to generate pairwise comparisons between means of individual sample groups and determine statistical significance (GraphPad Prism 7).
- The small molecule ML336 was recently discovered to have antiviral drug properties against VEEV [4]. While proven effective both in vivo and in vitro, it has limited solubility in aqueous solution, necessitating a delivery vehicle to improve drug stability and enable controlled release. Thus, we utilized a hybrid liposome-mesoporous silica nanoparticle technology that takes advantage of the loading capabilities and uniformity of MSNs and the biocompatibility and retention capabilities of liposomes in one drug delivery platform (
FIG. 1C ) [19, 22, 26, 27]. Referred to as LC-MSNs (see, e.g. [22]), these particles have the potential to protect and control the release of ML336 as well as be modified for tissue-specific targeting in future iterations of the LC-MSN technology [19, 20]. - LC-MSNs formation was modified based on past methods [19, 20]. First, monosized sub-150 nm MSNs were produced by up-scaling previous synthesis protocols. Optimized large batch synthesis procedures yielded highly homogeneous nanoparticles without alteration in structure or size. As shown by transmission electron microscopy (TEM) (
FIG. 3A ), the nanoparticles of approximately 75 nm were narrow in size distribution and displayed a hexagonal porous structure (FIG. 3A , inset). Their homogeneous colloidal size was also confirmed by DLS (95.9±2.1 nm, PDI=0.07±0.01) (Table 1). - Additionally, scanning electron microscopy (SEM) analysis (
FIG. 3B ) was performed in order to highlight the 3D hexagonal shape of the MSNs and, importantly, the open porous structure. As demonstrated inFIG. 3B (insert) andFIG. 3G-3I , surface accessible pores were clearly observed. In addition to SEM observation, N2 sorption also provided evidence on the pore shape and its surface accessibility. The resulting isotherm (FIG. 3I ) showed a steep increase in adsorption characteristic of a capillary condensation in mesopores capillary evaporation on the desorption branch, supporting the presence of uniform cylindrical mesopore open at both ends (surface-accessible). Accessibility of the pores was indirectly confirmed by the high (BET) accessible surface area found for these MSNs. Furthermore, 100 pore diameters were measured on the SEM micrograph; and their average was found to be 2.65±0.29 nm, which is in the same order of magnitude of the average pore size found by N2 sorption using DFT theory (˜3.5 nm) (FIG. 3G-3I ). - Liposomes were composed of 77.5:20:2.5 DSPC:Cholestrol:DSPE-PEG (2000) (mol %), a lipid composition chosen to ensure formation of a stable bilayer and to enhance colloidal stability of the resulting LC-MSNs. The primary lipid component, DSPC, was chosen due to its saturated acyl chain, as previous work has indicated that unsaturated lipids may contribute to reduced colloidal stability of LCMSNs over time [19]. Cholesterol was used to improve control over bilayer fluidity and leakage [22, 28], and a pegylated DSPE was included to increase circulation time and reduce protein adsorption to the LC-MSN surface [19, 22, 26, 27]. To assemble LC-MSNs, liposomes were applied to MSNs under sonication at a 5:1 liposomes:MSN (mass ratio), a similar ratio to those used previously (2:1-4:1) to produce high quality LC-MSNs [19]. Fusion between the negatively charged MSN and the zwitterionic liposome occurs due to electrostatic interactions and the lipophilic nature of the MSNs [15, 19, 29, 30].
- To ensure successful formation of LC-MSNs, particles were evaluated for size, charge, and morphology. In previous studies, increased particle size upon addition of a lipid bilayer to MSNs has been observed and is indicative of successful bilayer formation [19, 21, 28, 31, 32]. Results were similar in this work, as application of the lipid bilayer increased MSN particle diameter from ˜96 nm to ˜150 nm and 164 nm for unloaded and ML336 loaded particles, respectively, while maintaining a low PDI and thus good uniformity (Table 1,
FIG. 3A-3C ). -
TABLE 1 Z-average diameter, PDI, and zeta potential for MSNs, liposomes, unloaded LC-MSNs, loaded MSNs, and loaded LC-MSNs Z-average Zeta Potential Particle Type Medium Diameter (nm) PDI (mV) MSN Water 95.9 ± 2.1 0.072 ± 0.01 −25.0 ± 0.42 Liposome 50:50 125.03 ± 2.1 0.090 ± 0.01 −0.78 ± 0.20 PBS:Water Unloaded LC-MSN PBS 149.5 ± 1.5 0.116 ± 0.01 −0.263 ± 0.41 Loaded MSN PBS 487.4 ± 108 0.351 ± 0.06 NA Loaded LC-MSN PBS 164 ± 1.5 0.144 ± 0.02 −1.76 ± 0.26 - A neutralized surface charge of particles is another measure of successful MSN-liposome fusion [31, 33], as a reduction of MSN surface charge is observed due to charge shielding of deprotonated silanols on the MSN surface by the zwitterionic lipid bilayer [19]. Here, uncoated MSNs had zeta potential of −25.0±0.42 mV, similar to what has been reported in literature previously [13, 19, 20], which increased to nearly neutral levels when coated with a lipid bilayer (−0.263±0.41 mV, Table 1).
- As a final confirmation of successful bilayer application, a uniform lipid bilayer was observed in cryo-EM images (
FIG. 3C-3D ). Analysis of the cryo-EM images indicated a LC-MSN diameter of 88.1±11.8 nm and 86.5±12.0 nm and a bilayer thickness of 6.0±0.94 nm and 5.4±0.91 nm for loaded and unloaded LC-MSNs, respectively. The smaller diameter of the LC-MSNs determined by cryo-EM compared to DLS is consistent with previous reports [19]. The larger diameter observed in loaded LC-MSNs via DLS size analysis (Table 1) suggests the surface adsorbed ML336 may affect the hydrodynamic radius of the particle due to changes in surface hydrophobicity. Overall, ML336-loaded LC-MSNs were successful fabricated. - While MSNs are highly advantageous for small molecule delivery [12-15], aggregation of MSNs without surface modification or external coatings is commonly observed in high ionic strength physiologically relevant media due to a reduction in the Debye length and correspondingly the degree of electrostatic repulsion [12, 17, 20]. As might be predicted, ML336 loaded MSNs that were not coated with a lipid bilayer aggregated immediately in PBS (Table 1,
FIG. 4A ). In contrast, loaded LC-MSNs maintained colloidal stability for at least four days (FIG. 4A ), indicating their utility in both in vitro and in vivo applications. Taken as a whole, zeta-potential, cryo-EM and stability studies indicate the formation of a complete, conformal and uniform lipid bilayer on ML336 loaded LC-MSNs. - ML336 loading in LC-MSNs was determined to be about 20 μg ML336/mg LC-MSN, as measured by subtracting the amount of ML336 lost in the post-lipid-coating and loading washes from the total mass of ML336 loaded (
FIG. 4B-4D ). A linear burst release of ML336 was observed to occur in the first 4 hours, with little additional release thereafter (FIG. 3E-3F ). Overall, LC-MSNs released about 6.6 μg ML336/mg LC-MSNs in 24 hours, which correlated to 34% release of ML336 loaded (FIG. 3E-3F ; Table 2). No additional release was observed after 4 additional days. -
TABLE 2 Summary of ML336 Release from LC-MSNs Total ML336 Total ML336 Loaded (μg ML336/ % ML336 Released (μg ML336/ % ML336 mg LC-MSN) Loaded mg LC-MSN) Released 20 ± 3.4 20 ± 3.4 6.6 ± 1.3 33.5 ± 6.6 - Similar release was observed when LC-MSNs were incubated in
PBS pH 5, which mimics the intracellular endosome, while about 100% release was observed when LC-MSNs were incubated in methanol. The ML336 release observed here was similar to small molecule release from lipid coated MSNs in previous studies, where 0-35% release of loaded cargo was observed in ˜10 hrs atpH 7 for several different lipid bilayer compositions [19, 21, 31-33]. When the pH was dropped to 5, no additional release was observed, confirming what has been observed for a similar lipid bilayer composition previously [19]. In other reports where additional and sometimes nearly complete release of cargo from lipid-coated MSNs has been reported at low pH [32,33], specific acid-sensitive lipids have been employed to promote cargo release under acidic conditions. While still under investigation, the technology presented here could be modified to be acid-sensitive by adjusting the lipid composition of the lipid bilayer. However, the limited release at low pH observed in these studies could be beneficial, as it minimizes premature release and degradation of cargo in the endosomal compartment. - Complete ML336 release was observed when LC-MSNs were incubated in methanol, which is expected to disrupt the lipid bilayer and effectively extract ML336 from the MSN (
FIG. 4E-4F ). This result suggests that the limited solubility of ML336 in aqueous solution could prevent its release from the MSN core even after the expected disruption of the lipid bilayer at pH 5 [33]. Previously, MSNs loaded with hydrophobic drugs have shown reduced wettability, possibly retarding or preventing drug release prior to self-erosion of the silica matrix [33]. As degradation of the silica matrix is highly dependent upon surface functionalization, loaded cargo, relative concentration of particles, and the surrounding environment [34, 35], cargo release from LC-MSNs will be dependent upon specific conditions in both in vitro and in vivo environments. To enhance loading and release in future iterations of this technology, the MSN surface could be modified to optimize interactions between the MSN and ML336 [14, 15, 36]. The hydrophobicity of ML336 requires loading in a non-polar solvent (DMSO was used in these studies), while release occurs in physiological conditions (buffered aqueous solutions). As different properties dictate the interactions between ML336 and MSNs in aqueous versus non-aqueous solvents, it may be possible to maximize MSN-ML336 interactions in DMSO to enhance loading while minimizing MSN-ML336 interactions in PBS (or other aqueous solutions) to enhance release [36]. Overall, the results presented here indicate successful loading and release of ML336 from uniform LC-MSNs of high colloidal stability, providing an excellent prototype for future optimization and additional analysis in in vitro and in vivo studies. - To evaluate the performance of ML336-loaded LC-MSNs in vitro, their ability to inhibit virus in infected HeLa cells was assessed. First, a baseline was determined using soluble ML336, which inhibited TC-83 virus in a dose-dependent manner on HeLa cells with an IC-50 of 163 nM at 24 hrs (
FIG. 5A-5B ). The wildtype VEEV, a BSL-3 agent, was inhibited by ML336 in a similar manner (FIG. 5C-5D ) but was not used in subsequent studies due to associated high risk. The inhibition of VEEV by ML336 observed here was similar to previous studies [4]. - Cytotoxicity of ML336-loaded and unloaded LC-MSNs was assessed with HeLa cells. No visible differences in viability at 48 hrs was observed via LIVE/DEAD staining (
FIG. 6 ), in line with the high biocompatibility observed in cells treated with LC-MSNs previously [19, 33, 37] and with the limited toxicity observed when cells are treated with MSNs at a concentration less than 100 μg/mL [13]. To determine if ML336-loaded LC-MSNs inhibited virus, HeLa cells infected with the TC-83 virus were treated with ML336-loaded and unloaded LC-MSNs. Similar to soluble ML336, ML336-loaded LC-MSNs also inhibited virus in a dose-dependent manner, indicating that total ML336 release is proportional to LC-MSN mass and providing a method to tune drug dosage in a facile manner (FIG. 7 ). ML336 loaded LC-MSNs significantly decreased viral load by at least 4 orders of magnitude after 24 hrs and 6 orders of magnitude after 48 and 72 hrs (FIG. 8A-8B ), a greater reduction than previously observed for other small molecule VEEV inhibitors [9, 11] and similar to what has been observed for soluble ML336 [4]. Overall, these results indicate that ML336 loaded LCMSNs can successfully inhibit VEEV. - As discussed above, little to no additional ML336 release from LC-MSNs incubated in PBS was observed after four hours (
FIG. 3E-3F ). However, release of hydrophobic ML336 could depend heavily on the local microenvironment, especially if partially controlled by silica degradation [34, 35]. In order to evaluate if LC-MSNs were effective after the initial 4 hour burst release in vitro, ML336 loaded LC-MSNs were incubated in Opti-MEM for 4 hours, called “Pre-released LC-MSNs”, and then separated from the supernatant. TC-83 infected HeLa cells were then treated with LC-MSN supernatant and pre-released LC-MSNs and compared to cells treated with loaded LC-MSNs and untreated cells. LC-MSN supernatant inhibited virus at a similar level to ML336 loaded LC-MSNs until the 72 hr timepoint, at which point loaded LC-MSNs inhibited virus to a greater extent (FIG. 8C ,FIG. 9A-9D ). This indicates that while released ML336 remains bioactive, it is possible that LC-MSNs protect ML336 over time and/or continually release additional ML336 in a manner different from our test tube release studies. - While pre-released LC-MSNs inhibited virus in a similar manner to loaded LC-MSNs and LC-MSN supernatant at 24 hrs, by 48 hrs the extent of viral inhibition was significantly lower than cells treated with loaded LC-MSNs or LC-MSN supernatant. By 72 hrs, pre-released LC-MSNs showed no additional inhibition as compared to cells with no treatment (
FIG. 8C ,FIG. 9A-9D ). This indicates that LC-MSNs release additional ML336 after the initial four hour burst release, which may either be undetectable in the loading and release studies or does not occur prior to cell internalization and disruption of the lipid bilayer [33]. Taken as a whole, this data suggests that release from LC-MSNs occurs for longer than four hrs (possibly up to 48 hrs) and may depend on intracellular uptake. In addition, these studies were reproducible across multiple studies that employed different batches of particles (FIG. 9A-9D ), indicating the robustness of the technology as a whole. - To begin to understand the dependency of LC-MSN cellular internalization on lipid bilayer disruption and complete drug cargo release, we first investigated whether LC-MSNs enter cells through endocytosis. LC-MSNs conjugated with affinity ligands are known to enter cells using trafficking pathways of the targeting receptor. For example, cholera toxin B conjugated LC-MSNs use caveolin-mediated endocytosis for internalization after binding the GM1 ganglioside receptor [20]. LC-MSNs have also been formulated to avoid non-specific uptake in blood circulation [19]. However, LC-MSN uptake in static conditions represented in these studies is not well understood.
- To determine whether LC-MSNs undergo cellular internalization through endocytosis, fluorescent LC-MSNs containing Cy3-labeled MSN cores were used to facilitate visualization and quantitation of entry into HeLa cells while in the presence of various endocytosis inhibitors. Hela cells were treated with a pH dependent endocytosis inhibitor (bafilomycin, BAF), clathrin-mediated endocytosis inhibitors (dynamin II inhibitor dynasore, DYN, and chlorpromazine, CPZ), caveolae-medicated endocytosis inhibitors (phorbol 12-myristate 13-acetate, PMA and DYN), or macropinocytosis inhibitors (wortmannin, wort, and p21-activated kinase inhibitor III, IPA-3) for 1 hr prior to the addition of Cy3-labeled LC-MSNs. Cells were vigorously washed to remove free particles and then examined by microscopy methods. Cy3-labeled LC-MSNs were readily internalized by Hela cells under untreated or no inhibitor (NI) conditions. LC-MSNs were also clearly inhibited in the presence of BAF, CPZ, and DYN, but not by inhibitors of macropinocytosis (wort, IPA-3) nor caveolin-mediated endocytosis (PMA), suggesting the role of clathrin-mediated endocytosis in cellular internalization of LC-MSNs (
FIG. 10A ). - To quantify the results obtained by microscopy, flow cytometry was used to measure the percentage of internalized Cy3-labeled LC-MSNs. HeLa cells were treated with the panel of endocytosis inhibitors for 1 hr prior to and during incubation with fluorescent LC-MSNs. Before flow cytometry analysis, the cells were washed to remove unbound particles. Again, inhibitors of endosomal acidification (BAF) almost completely inhibited LC-MSN internalization, while those of clathrin-mediated endocytosis dramatically reduced LC-MSN uptake. The percentage of cells that internalized LC-MSNs was reduced by 71% with DYN, and 84% with CPZ (
FIG. 10B and FIG. 11A) as compared to untreated control conditions, thus correlating with microscopy data. As viruses commonly use endocytosis for cellular entry, we confirmed the specificity of these inhibitors with viruses known to enter HeLa cells via endocytosis using vesicular stomatitis virus for clathrin-mediated endocytosis or Rift Valley fever virus for caveolae-mediated endocytosis dependent entry (FIG. 11B-11C ) [38]. - To confirm the labeled LC-MSNs were internalized and not on the cell surface, high resolution confocal microscopy techniques were employed. HeLa cells were incubated with LC-MSNs for either 45 min or 20 hrs, washed, and then fixed for immunofluorescence staining. As shown in
FIG. 10C-10D , HeLa cells incubated with LC-MSNs for 20 hrs were internalized as indicated by 3D rendering of LC-MSNs with actin, microtubules, and nuclei intracellular markers. An actin stain was used to mark the periphery of the cell as actin filaments are concentrated at the cell periphery and form a 3D network that determines cell shape. Microtubule labeling using tubulin antibodies provided another reference for intracellular localization and depth of LC-MSNs within the cell. LC-MSNs were visualized as beneath actin filaments, on the same plane as the microtubules, and above the cell nucleus. Furthermore, these data indicated a time dependent mechanism of entry as particles were not seen intracellularly at 45 min (FIG. 10C-10D ). Taken together, LC-MSNs enter cells through clathrin-mediated endocytosis and may provide a mechanism of additional drug/cargo release at the site of LC-MSN accumulation. Identifying the LC-MSNs cell entry pathway as clathrin-mediated endocytosis may provide a mechanism to design additional drug/cargo release at the site of LC-MSN accumulation. Overall, these results motivated a further investigation of the ability of ML336 loaded LC-MSNs to inhibit virus in vivo. - As with all nanoparticle-based systems, the potential for LC-MSNs to dissolve, aggregate, and interact with living cells and animal tissues is dependent upon properties specific to their unique composition [14]. In addition, the toxicity of MSNs and LC-MSNs in general has yet to be fully assessed and can vary depending upon size and surface properties [12, 13, 18]. Thus, prior to conducting animal studies to evaluate antiviral efficacy, a safety study was conducted to determine if the LC-MSNs developed in this work affected mouse weight and survival over fifteen days. Mice were injected with 1 mg LC-MSNs twice daily for four days, and all animals survived treatment with no significant differences in total animal weight between LC-MSN and PBS treated groups (
FIG. 12A ). Previously, MSNs have been seen to accumulate in the spleen, liver, bladder, and kidneys [13, 14]. In this work, no significant differences were observed between lung, liver, spleen, kidney, or brain weights in animals treated with LC-MSNs as compared to PBS only (FIG. 12B ). A lack of weight change in tissue where bioaccumulation was likely the highest (liver, spleen and kidney) further suggests a lack of LC-MSN toxicity. Similarly, tissues processed for histology revealed a normal morphology in brain, spleen, and kidney sections in LC-MSN treated mice while only very mild changes of some sections were seen in livers and lungs of nanoparticle dosed animals (FIG. 12C ). - In summary, we observed no toxicity when a 0.11 g LC-MSNs/kg mouse dose was administered each day for four days, resulting in a total possible accumulated mass of silica nanoparticles of 0.44 g/kg. This correlates well with previous work, where MSN toxicity in mice was observed to be problematic when MSN were administered one time at 1.2 g/kg by IV injection [39], but little to no toxicity was observed when 0.2 g/kg was administered once a day for 10 days [40]. The nanoparticles used in the work presented here also include the addition of the lipid bilayer, likely improving biocompatibility, increasing circulation time, and reducing toxicity as compared to uncoated MSNs [12]. Overall, results indicated that administration of LC-MSNs did not cause significant toxicity in mice, motivating further studies to investigate the ability of drug-loaded particles to inhibit viral infection.
- In the first set of animal studies, mice were treated with 1 mg loaded LC-MSNs, unloaded LC-MSNs, free ML336, or PBS only. Mice treated with ML336 loaded LC-MSNs showed greater survival than animals in the other three groups, though this result was not statistically significant (
FIG. 13A ). As we observed that LC-MSNs inhibited virus in a dose-dependent manner in in vitro studies (FIG. 7 ), we were interested to see if an increased LC-MSN dose would improve animal outcomes. Thus, in the second set of animal studies, animals were dosed with 1.5 mg LC-MSNs. No differences were observed in spleen viral load (FIG. 13C ), and very limited viral loads were detected in livers, kidneys, or serum (FIG. 14A-14C ), similar to what has been observed in past studies characterizing TC-83 intranasal infection in C3H/HeN mice [8]. - However, viral load in the brain was significantly reduced by about 10-fold in the ML336 loaded LC-MSN treated mice as compared to the PBS treated animals after 4 days (
FIG. 13B ). As viral load in mice treated with free ML336 was not significantly different from PBS treated animals, LC-MSNs may protect ML336 or increase circulation time in a manner that permits enhanced antiviral activity. In the future, a larger number of animals will help further elucidate trends. Overall, these results are encouraging and indicate the potential utility of ML336-loaded LC-MSNs in inhibiting VEEV infection. - No FDA approved therapeutics are available for VEEV [3, 4], though several studies have highlighted the ability of small molecules to inhibit VEEV both in vitro and in vivo [6-11]. Small molecule inhibitors of VEEV has been moderately successful, though drug toxicity has remained an issue [7, 10]. In addition, similar to the results in this study (
FIG. 12B ), animals treated with small molecule VEEV inhibitors show less than 10-fold brain viral titer reduction as compared to untreated control groups [7, 11], which may result in increased neurological impairment. - Future iterations of the LC-MSNs presented in this work have the potential to improve these outcomes by reducing toxicity and enabling targeting specific to the blood-brain barrier in the case of VEEV infections. First, LC-MSNs can prevent toxicity by 1) reducing the concentration required for drug efficacy, both through improvements in drug solubility/stability as well as circulation time [12, 19], and 2) protecting the cellular microenvironment from harmful cargo prior to triggered release, either through rupture of the lipid bilayer or a specific chemically triggered mechanism [22]. Second, the LC-MSN lipid bilayer can be modified to specifically target a tissue of interest, such as the blood-brain barrier [20, 22]. LC-MSNs are particularly advantageous because properties of the lipid bilayer and the MSN can be independently tuned, enabling simultaneous optimization of the lipid bilayer for tissue-specific targeting and the core to maximize drug-specific loading. Overall, the studies presented here highlight the ability of drug loaded LC-MSNs to prevent viral infection in one particular case, but the versatility and modifiability of this technology will enable use of LC-MSNs to prevent viral infection in a variety of future applications.
- Here, we presented the first use of LC-MSNs to deliver ML336 for TC-83 VEEV inhibition both in vitro and in vivo. ML336 loaded LC-MSNs were successfully coated with a lipid bilayer, which significantly improved colloidal stability, and released cargo over the course of 4 hours. Viral loads were reduced by 4-6 orders of magnitude in TC-83 VEEV infected HeLa cells treated with ML336 loaded LC-MSNs, which was repeatable across several particle batches in different studies. Furthermore, in vitro studies could indicate the possibility of additional release of ML336 after cellular internalization via clathrin-mediated endocytosis and enhanced ML336 stability when loaded in LC-MSNs.
- Safety studies indicated that LC-MSNs were not toxic in mice at the doses administered in this study. In mice infected with TC-83 VEEV, animals treated with ML336 loaded LC-MSNs showed a significant reduction of viral load in the brain after four days of treatment. Overall, these studies highlight the utility of LC-MSNs for drug delivery in antiviral applications, and provide an additional defense against VEEV and other alphaviruses in the cases of natural infection or bioterrorism.
- 1. Atasheva S et al., Venezuelan equine encephalitis virus capsid protein inhibits nuclear import in Mammalian but not in mosquito cells. J. Virol. 2008; 82(8):4028-41.
- 2. Nagata L P et al., Vaccines and therapeutics for the encephalitic alphaviruses. Future Virol. 2013; 8(7):661-74.
- 3. Sidwell R W et al., Viruses of the Bunya-and Togaviridae families: potential as bioterrorism agents and means of control. Antiviral Res. 2003; 57(1-2):101-11.
- 4. Schroeder C E et al., Development of (E)-2-((1,4-dimethylpiperazin-2-ylidene)amino)-5-nitro-N-phenylbenzamide, ML336: Novel 2-amidinophenylbenzamides as potent inhibitors of Venezuelan equine encephalitis virus. J. Med. Chem. 2014; 57(20):8608-21.
- 5. Zacks M A and Paessler S, Encephalitic alphaviruses. Vet. Microbiol. 2010; 140(3-4):281-6.
- 6. Chung D H et al., Discovery of a novel compound with anti-venezuelan equine encephalitis virus activity that targets the
nonstructural protein 2. PLoS Pathog. 2014; 10(6):e1004213 (10 pp.). - 7. Julander J G et al., Treatment of Venezuelan equine encephalitis virus infection with (−)-carbodine. Antiviral Res. 2008; 80(3):309-15.
- 8. Julander J G et al., C3H/HeN mouse model for the evaluation of antiviral agents for the treatment of Venezuelan equine encephalitis virus infection. Antiviral Res. 2008; 78(3):230-41.
- 9. Kehn-Hall K et al., Modulation of GSK-3beta activity in Venezuelan equine encephalitis virus infection. PLoS One 2012; 7(4):e34761 (12 pp.).
- 10. Langsjoen R M et al., Host oxidative folding pathways offer novel anti-chikungunya virus drug targets with broad spectrum potential. Antiviral Res. 2017; 143:246-51.
- 11. Madsen C et al., Small molecule inhibitors of Ago2 decrease Venezuelan equine encephalitis virus replication. Antiviral Res. 2014; 112:26-37.
- 12. Tang F et al., Mesoporous silica nanoparticles: synthesis, biocompatibility and drug delivery. Adv. Mater. 2012; 24(12):1504-34.
- 13. Vivero-Escoto J L et al., Mesoporous silica nanoparticles for intracellular controlled drug delivery. Small 2010; 6(18):1952-67.
- 14. Mamaeva V et al., Mesoporous silica nanoparticles in medicine—recent advances. Adv. Drug Deliv. Rev. 2013; 65(5):689-702.
- 15. Slowing I I et al., Mesoporous silica nanoparticles as controlled release drug delivery and gene transfection carriers. Adv. Drug Deliv. Rev. 2008; 60(11):1278-88.
- 16. Li Z et al., Mesoporous silica nanoparticles in biomedical applications. Chem. Soc. Rev. 2012; 41(7):2590-605.
- 17. Lu J et al., Mesoporous silica nanoparticles as a delivery system for hydrophobic anticancer drugs. Small 2007; 3(8):1341-6.
- 18. Rosenholm J M et al., Towards multifunctional, targeted drug delivery systems using mesoporous silica nanoparticles—opportunities & challenges. Nanoscale 2010; 2(10):1870-83.
- 19. Durfee P N et al., Mesoporous silica nanoparticle-supported lipid bilayers (protocells) for active targeting and delivery to individual leukemia cells. ACS Nano 2016; 10(9):8325-45.
- 20. Gonzalez Porras M A et al., A novel approach for targeted delivery to motoneurons using cholera toxin-B modified protocells. J. Neurosci. Methods 2016; 273:160-74.
- 21. Cauda V et al., Colchicine-loaded lipid bilayer-coated 50 nm mesoporous nanoparticles efficiently induce microtubule depolymerization upon cell uptake. Nano Lett. 2010; 10(7):2484-92.
- 22. Butler K S et al., Protocells: modular mesoporous silica nanoparticle-supported lipid bilayers for drug delivery. Small 2016; 12(16):2173-85.
- 23. Bimbo L M et al., Inhibition of Influenza A virus infection in vitro by saliphenylhalamide-loaded porous silicon nanoparticles. ACS Nano 2013; 7(8):6884-93.
- 24. van Schooneveld M M et al., Improved biocompatibility and pharmacokinetics of silica nanoparticles by means of a lipid coating: a multimodality investigation. Nano Lett. 2008. 8(8): 2517-25.
- 25. Roggers R A et al., Chemically reducible lipid bilayer coated mesoporous silica nanoparticles demonstrating controlled release and HeLa and normal mouse liver cell biocompatibility and cellular internalization. Mol. Pharm. 2012; 9(9):2770-7.
- 26. Teng I T et al., Phosphohpid-functionalized mesoporous silica nanocarriers for selective photodynamic therapy of cancer. Biomaterials 2013; 34(30):7462-70.
- 27. Wang L S et al., Biofunctionalized phospholipid-capped mesoporous silica nanoshuttles for targeted drug delivery: improved water suspensibility and decreased nonspecific protein binding. ACS Nano 2010; 4(8):4371-9.
- 28. Tarn D et al., Mesoporous silica nanoparticle nanocarriers: biofunctionality and biocompatibility. Acc. Chem. Res. 2013; 46(3):792-801.
- 29. Mornet S et al., The formation of supported lipid bilayers on silica nanoparticles revealed by cryoelectron microscopy. Nano Lett. 2005; 5(2):281-5.
- 30. Liu J et al., Porous nanoparticle supported lipid bilayers (protocells) as delivery vehicles. J. Am. Chem. Soc. 2009; 131(4):1354-5.
- 31. Han N et al., Hybrid lipid-capped mesoporous silica for stimuli-responsive drug release and overcoming multidrug resistance. ACS Appl. Mater. Interfaces 2015; 7(5):3342-51.
- 32. Wang D et al., The eradication of breast cancer cells and stem cells by 8 hydroxyquinoline-loaded hyaluronan modified mesoporous silica nanoparticle-supported lipid bilayers containing docetaxel. Biomaterials 2013; 34(31):7662-73.
- 33. Zhang X et al., Biofunctionalized polymer-lipid supported mesoporous silica nanoparticles for release of chemotherapeutics in multidrug resistant cancer cells. Biomaterials 2014; 35(11):3650-65.
- 34. von Haartman E et al., On the intracellular release mechanism of hydrophobic cargo and its relation to the biodegradation behavior of mesoporous silica nanocarriers. Eur. J. Pharm. Sci. 2016; 95:17-27.
- 35. Braun K et al., Dissolution kinetics of mesoporous silica nanoparticles in different simulated body fluids. J. Sol-Gel Sci. Technol. 2016; 79(2):319-27.
- 36. Maleki A et al., Mesoporous silica materials: from physico-chemical properties to enhanced dissolution of poorly water-soluble drugs. J. Control. Release 2017; 262:329-47.
- 37. Han D H et al., Direct cellular delivery of human proteasomes to delay tau aggregation. Nat. Commun. 2014; 5:5633 (8 pp.).
- 38. Harmon B et al., Rift Valley fever virus strain MP-12 enters mammalian host cells via caveola-mediated endocytosis. J. Virol. 2012; 86:12954-70.
- 39. Riikonen J et al., Systematic in vitro and in vivo study on porous silicon to improve the oral bioavailability of celecoxib. Biomaterials 2015; 52:44-55.
- 40. Lu J et al., Biocompatibility, biodistribution, and drug-delivery efficiency of mesoporous silica nanoparticles for cancer therapy in animals. Small 2010; 6(16):1794-805.
- 41. Townson J L et al., Re-examining the size/charge paradigm: differing in vivo characteristics of size-and charge-matched mesoporous silica nanoparticles. J. Am. Chem. Soc. 2013; 135:16030-3.
- 42. Lin Y S et al., Stability of small mesoporous silica nanoparticles in biological media. Chem. Commun. (Camb.) 2011; 47:532-4.
- 43. Sherman M B et al., Stability of cucumber necrosis virus at the quasi-6-fold axis affects zoospore transmission. J. Virol. 2017; 91:1-12.
- All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each independent publication or patent application was specifically and individually indicated to be incorporated by reference.
- While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims.
- Other embodiments are within the claims.
Claims (20)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/306,452 US20210330809A1 (en) | 2018-06-22 | 2021-05-03 | Lipid-coated particles for treating viral infections |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862689037P | 2018-06-22 | 2018-06-22 | |
US16/443,316 US11045554B1 (en) | 2018-06-22 | 2019-06-17 | Lipid-coated particles for treating viral infections |
US17/306,452 US20210330809A1 (en) | 2018-06-22 | 2021-05-03 | Lipid-coated particles for treating viral infections |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/443,316 Division US11045554B1 (en) | 2018-06-22 | 2019-06-17 | Lipid-coated particles for treating viral infections |
Publications (1)
Publication Number | Publication Date |
---|---|
US20210330809A1 true US20210330809A1 (en) | 2021-10-28 |
Family
ID=76548174
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/443,316 Active US11045554B1 (en) | 2018-06-22 | 2019-06-17 | Lipid-coated particles for treating viral infections |
US17/306,452 Abandoned US20210330809A1 (en) | 2018-06-22 | 2021-05-03 | Lipid-coated particles for treating viral infections |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/443,316 Active US11045554B1 (en) | 2018-06-22 | 2019-06-17 | Lipid-coated particles for treating viral infections |
Country Status (1)
Country | Link |
---|---|
US (2) | US11045554B1 (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11517539B2 (en) | 2016-02-15 | 2022-12-06 | University Of Georgia Research Foundation, Inc. | IPA-3-loaded liposomes and methods of use thereof |
EP3663257A1 (en) * | 2018-12-05 | 2020-06-10 | Fundació Institut de Bioenginyeria de Catalunya (IBEC) | Functionalized enzyme-powered nanomotors |
WO2023183845A1 (en) * | 2022-03-22 | 2023-09-28 | West Pharmaceutical Services, Inc. | Liposome lubricant composition and preparation |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140079774A1 (en) * | 2011-04-28 | 2014-03-20 | Stc.Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery and methods of using same |
Family Cites Families (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8734816B2 (en) | 2009-01-05 | 2014-05-27 | Stc.Unm | Porous nanoparticle supported lipid bilayer nanostructures |
US8992984B1 (en) | 2009-10-21 | 2015-03-31 | Stc.Unm | Protocells and their use for targeted delivery of multicomponent cargos to cancer cells |
WO2011097607A1 (en) | 2010-02-08 | 2011-08-11 | Southern Research Institute | Anti-viral treatment and assay to screen for anti-viral agent |
SG11201401499XA (en) | 2011-10-14 | 2014-09-26 | Stc Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery including transdermal delivery of cargo and methods thereof |
WO2013103614A1 (en) | 2011-12-30 | 2013-07-11 | Stc.Unm | Crlf-2 binding peptides, protocells and viral-like particles useful in the treatment of cancer, including acute lymphoblastic leukemia (all) |
PT2898075E (en) | 2012-12-12 | 2016-06-16 | Harvard College | Engineering and optimization of improved systems, methods and enzyme compositions for sequence manipulation |
WO2014093869A1 (en) | 2012-12-13 | 2014-06-19 | University Of Kansas | 6-substituted quinazolinone inhibitors |
WO2014165608A1 (en) | 2013-04-02 | 2014-10-09 | Stc. Unm | Antibiotic protocells and related pharmaceutical formulations and methods of treatment |
WO2014165617A1 (en) | 2013-04-02 | 2014-10-09 | Stc.Unm | Mesoporous alum nanoparticles as universal platform for antigen adsorption, presentation, and delivery |
US9855217B2 (en) | 2013-09-18 | 2018-01-02 | Stc. Unm | Toroidal mesoporous silica nanoparticles (TMSNPs) and related protocells |
US20160287717A1 (en) | 2013-09-18 | 2016-10-06 | Stc.Unm | Core and Surface Modification of Mesoporous Silica Nanoparticles to Achieve Cell Specific Targeting In Vivo. |
US20160090603A1 (en) | 2014-09-30 | 2016-03-31 | Sandia Corporation | Delivery platforms for the domestication of algae and plants |
US20180110831A1 (en) | 2015-03-09 | 2018-04-26 | Stc.Unm | Cd 47 containing porous nanoparticle supported lipid bilayers (protocells) field of the invention |
US20180105430A1 (en) | 2015-03-11 | 2018-04-19 | Stc.Unm | Generation of mesoporous materials using multiphase surfactant systems |
WO2017023407A2 (en) | 2015-06-01 | 2017-02-09 | Stc. Unm | Nanobiocomposite compositions and methods |
WO2017041033A1 (en) | 2015-09-04 | 2017-03-09 | Brinker C Jeffrey | Mesoporous silica nanoparticles and supported lipid bi-layer nanoparticles for biomedical applications |
WO2017041032A1 (en) | 2015-09-04 | 2017-03-09 | Brinker C Jeffrey | Protocells to treat microbial infection and for synergistic delivery |
US11672866B2 (en) | 2016-01-08 | 2023-06-13 | Paul N. DURFEE | Osteotropic nanoparticles for prevention or treatment of bone metastases |
-
2019
- 2019-06-17 US US16/443,316 patent/US11045554B1/en active Active
-
2021
- 2021-05-03 US US17/306,452 patent/US20210330809A1/en not_active Abandoned
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140079774A1 (en) * | 2011-04-28 | 2014-03-20 | Stc.Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery and methods of using same |
Non-Patent Citations (1)
Title |
---|
Schroeder et al., J. Med. Chem., 2014, 57(20), pages 8608-8621. (Year: 2014) * |
Also Published As
Publication number | Publication date |
---|---|
US11045554B1 (en) | 2021-06-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20210330809A1 (en) | Lipid-coated particles for treating viral infections | |
Yang et al. | Drug delivery via cell membrane fusion using lipopeptide modified liposomes | |
Nie et al. | Cancer-cell-membrane-coated nanoparticles with a yolk–shell structure augment cancer chemotherapy | |
Cardarelli et al. | Cholesterol-dependent macropinocytosis and endosomal escape control the transfection efficiency of lipoplexes in CHO living cells | |
Wang et al. | Facile synthesis of uniform virus-like mesoporous silica nanoparticles for enhanced cellular internalization | |
Luo et al. | Systematic evaluation of transferrin-modified porous silicon nanoparticles for targeted delivery of doxorubicin to glioblastoma | |
Galiana et al. | Preclinical antitumor efficacy of senescence-inducing chemotherapy combined with a nanoSenolytic | |
LaBauve et al. | Lipid-coated mesoporous silica nanoparticles for the delivery of the ML336 antiviral to inhibit encephalitic alphavirus infection | |
Meng et al. | Aspect ratio determines the quantity of mesoporous silica nanoparticle uptake by a small GTPase-dependent macropinocytosis mechanism | |
Durfee et al. | Mesoporous silica nanoparticle-supported lipid bilayers (protocells) for active targeting and delivery to individual leukemia cells | |
US10624852B2 (en) | Liposomes comprising a calcium phosphate-containing precipitate | |
Bimbo et al. | Inhibition of influenza A virus infection in vitro by saliphenylhalamide-loaded porous silicon nanoparticles | |
Meng et al. | Engineered design of mesoporous silica nanoparticles to deliver doxorubicin and P-glycoprotein siRNA to overcome drug resistance in a cancer cell line | |
JP5863670B2 (en) | Synthetic nanostructures containing nucleic acids and / or other components | |
Kong et al. | Virucidal nano-perforator of viral membrane trapping viral RNAs in the endosome | |
US20110117026A1 (en) | Methods and compositions for the delivery of bioactive compounds | |
Garg et al. | pH-sensitive PEGylated liposomes functionalized with a fibronectin-mimetic peptide show enhanced intracellular delivery to colon cancer cells | |
Zhang et al. | Receptor-mediated surface charge inversion platform based on porous silicon nanoparticles for efficient cancer cell recognition and combination therapy | |
Li et al. | Stepwise-acid-active multifunctional mesoporous silica nanoparticles for tumor-specific nucleus-targeted drug delivery | |
JP2014532071A (en) | Lipid bilayer (protocell) supported on porous nanoparticles for targeted delivery including transdermal delivery of cargo and method thereof | |
Wang et al. | Surface-modified nanoerythrocyte loading DOX for targeted liver cancer chemotherapy | |
Zheng et al. | Dual-targeting multifuntional mesoporous silica nanocarrier for codelivery of siRNA and ursolic acid to folate receptor overexpressing cancer cells | |
Song et al. | Mechanisms of transcellular transport of wheat germ agglutinin-functionalized polymeric nanoparticles in Caco-2 cells | |
Deng et al. | Transmembrane pathways and mechanisms of rod-like paclitaxel nanocrystals through MDCK polarized monolayer | |
Yang et al. | Binary‐copolymer system base on low‐density lipoprotein‐coupled N‐succinyl chitosan lipoic acid micelles for co‐delivery MDR1 siRNA and paclitaxel, enhances antitumor effects via reducing drug |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
AS | Assignment |
Owner name: UNM RAINFOREST INNOVATIONS, NEW MEXICO Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:THE REGENTS OF THE UNIVERSITY OF NEW MEXICO;REEL/FRAME:059378/0502 Effective date: 20211209 Owner name: UNM RAINFOREST INNOVATIONS, NEW MEXICO Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:BRINKER, CHARLES JEFFREY;REEL/FRAME:059378/0480 Effective date: 20210517 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: FINAL REJECTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |