WO2021261998A1 - Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof - Google Patents
Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof Download PDFInfo
- Publication number
- WO2021261998A1 WO2021261998A1 PCT/NL2021/050394 NL2021050394W WO2021261998A1 WO 2021261998 A1 WO2021261998 A1 WO 2021261998A1 NL 2021050394 W NL2021050394 W NL 2021050394W WO 2021261998 A1 WO2021261998 A1 WO 2021261998A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- saponin
- xyl
- rha
- group
- fuc
- Prior art date
Links
- 150000007949 saponins Chemical class 0.000 title claims abstract description 166
- 229930182490 saponin Natural products 0.000 title claims abstract description 155
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims abstract description 140
- 230000001225 therapeutic effect Effects 0.000 title description 16
- 210000004027 cell Anatomy 0.000 claims abstract description 206
- 239000012636 effector Substances 0.000 claims abstract description 150
- OVRNDRQMDRJTHS-KEWYIRBNSA-N N-acetyl-D-galactosamine Chemical group CC(=O)N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-KEWYIRBNSA-N 0.000 claims abstract description 111
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 60
- 239000003446 ligand Substances 0.000 claims abstract description 53
- 230000014509 gene expression Effects 0.000 claims abstract description 49
- 238000000034 method Methods 0.000 claims abstract description 45
- 101150075175 Asgr1 gene Proteins 0.000 claims abstract description 43
- -1 TTR Proteins 0.000 claims abstract description 40
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 25
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 24
- 238000011282 treatment Methods 0.000 claims abstract description 24
- 239000003814 drug Substances 0.000 claims abstract description 23
- 201000010099 disease Diseases 0.000 claims abstract description 21
- 102100030755 5-aminolevulinate synthase, nonspecific, mitochondrial Human genes 0.000 claims abstract description 20
- 102100038955 Proprotein convertase subtilisin/kexin type 9 Human genes 0.000 claims abstract description 20
- 210000000172 cytosol Anatomy 0.000 claims abstract description 20
- 101150102415 Apob gene Proteins 0.000 claims abstract description 19
- 238000000338 in vitro Methods 0.000 claims abstract description 18
- 230000001404 mediated effect Effects 0.000 claims abstract description 16
- 101000843649 Homo sapiens 5-aminolevulinate synthase, nonspecific, mitochondrial Proteins 0.000 claims abstract description 15
- 101150010882 S gene Proteins 0.000 claims abstract description 15
- 101150003160 X gene Proteins 0.000 claims abstract description 15
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 13
- 201000011510 cancer Diseases 0.000 claims abstract description 13
- 230000000295 complement effect Effects 0.000 claims abstract description 12
- 238000011321 prophylaxis Methods 0.000 claims abstract description 10
- 230000005802 health problem Effects 0.000 claims abstract description 7
- 108700016481 Acute Hepatic Porphyria Proteins 0.000 claims abstract description 6
- 208000003914 Acute hepatic porphyria Diseases 0.000 claims abstract description 6
- 208000023275 Autoimmune disease Diseases 0.000 claims abstract description 6
- 208000035473 Communicable disease Diseases 0.000 claims abstract description 6
- 201000003542 Factor VIII deficiency Diseases 0.000 claims abstract description 6
- 208000034846 Familial Amyloid Neuropathies Diseases 0.000 claims abstract description 6
- 208000035150 Hypercholesterolemia Diseases 0.000 claims abstract description 6
- 208000004777 Primary Hyperoxaluria Diseases 0.000 claims abstract description 6
- 208000036142 Viral infection Diseases 0.000 claims abstract description 6
- 206010002022 amyloidosis Diseases 0.000 claims abstract description 6
- 208000009429 hemophilia B Diseases 0.000 claims abstract description 6
- 208000033552 hepatic porphyria Diseases 0.000 claims abstract description 6
- 208000002672 hepatitis B Diseases 0.000 claims abstract description 6
- 208000015181 infectious disease Diseases 0.000 claims abstract description 6
- 208000019423 liver disease Diseases 0.000 claims abstract description 6
- 201000007905 transthyretin amyloidosis Diseases 0.000 claims abstract description 6
- 230000009385 viral infection Effects 0.000 claims abstract description 6
- 150000001720 carbohydrates Chemical group 0.000 claims description 111
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 claims description 97
- 108091034117 Oligonucleotide Proteins 0.000 claims description 89
- FHICGHSMIPIAPL-HDYAAECPSA-N [2-[3-[6-[3-[(5R,6aS,6bR,12aR)-10-[6-[2-[2-[4,5-dihydroxy-3-(3,4,5-trihydroxyoxan-2-yl)oxyoxan-2-yl]ethoxy]ethyl]-3,4,5-trihydroxyoxan-2-yl]oxy-5-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carbonyl]peroxypropyl]-5-[[5-[8-[3,5-dihydroxy-4-(3,4,5-trihydroxyoxan-2-yl)oxyoxan-2-yl]octoxy]-3,4-dihydroxy-6-methyloxan-2-yl]methoxy]-3,4-dihydroxyoxan-2-yl]propoxymethyl]-5-hydroxy-3-[(6S)-6-hydroxy-2,6-dimethylocta-2,7-dienoyl]oxy-6-methyloxan-4-yl] (2E,6S)-6-hydroxy-2-(hydroxymethyl)-6-methylocta-2,7-dienoate Chemical compound C=C[C@@](C)(O)CCC=C(C)C(=O)OC1C(OC(=O)C(\CO)=C\CC[C@](C)(O)C=C)C(O)C(C)OC1COCCCC1C(O)C(O)C(OCC2C(C(O)C(OCCCCCCCCC3C(C(OC4C(C(O)C(O)CO4)O)C(O)CO3)O)C(C)O2)O)C(CCCOOC(=O)C23C(CC(C)(C)CC2)C=2[C@@]([C@]4(C)CCC5C(C)(C)C(OC6C(C(O)C(O)C(CCOCCC7C(C(O)C(O)CO7)OC7C(C(O)C(O)CO7)O)O6)O)CC[C@]5(C)C4CC=2)(C)C[C@H]3O)O1 FHICGHSMIPIAPL-HDYAAECPSA-N 0.000 claims description 67
- 239000004055 small Interfering RNA Substances 0.000 claims description 66
- 102000005427 Asialoglycoprotein Receptor Human genes 0.000 claims description 65
- 108010006523 asialoglycoprotein receptor Proteins 0.000 claims description 65
- 108020004459 Small interfering RNA Proteins 0.000 claims description 62
- 230000015572 biosynthetic process Effects 0.000 claims description 57
- 230000009466 transformation Effects 0.000 claims description 48
- 238000006243 chemical reaction Methods 0.000 claims description 47
- 108700012359 toxins Proteins 0.000 claims description 45
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 claims description 44
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 claims description 44
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 claims description 44
- 239000003053 toxin Substances 0.000 claims description 43
- 231100000765 toxin Toxicity 0.000 claims description 43
- 102000004169 proteins and genes Human genes 0.000 claims description 37
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 35
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical group OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 claims description 32
- 150000007523 nucleic acids Chemical class 0.000 claims description 32
- 230000030279 gene silencing Effects 0.000 claims description 31
- 102000039446 nucleic acids Human genes 0.000 claims description 31
- 108020004707 nucleic acids Proteins 0.000 claims description 31
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 29
- 241000700721 Hepatitis B virus Species 0.000 claims description 28
- 125000003172 aldehyde group Chemical group 0.000 claims description 28
- 230000008685 targeting Effects 0.000 claims description 28
- 125000003668 acetyloxy group Chemical group [H]C([H])([H])C(=O)O[*] 0.000 claims description 27
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 claims description 25
- 239000002679 microRNA Substances 0.000 claims description 25
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims description 25
- 102000015395 alpha 1-Antitrypsin Human genes 0.000 claims description 24
- 108010050122 alpha 1-Antitrypsin Proteins 0.000 claims description 24
- 229940024142 alpha 1-antitrypsin Drugs 0.000 claims description 24
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 claims description 24
- 108010071690 Prealbumin Proteins 0.000 claims description 23
- 102000009190 Transthyretin Human genes 0.000 claims description 23
- 238000001212 derivatisation Methods 0.000 claims description 23
- 108010045100 HSP27 Heat-Shock Proteins Proteins 0.000 claims description 22
- 102100039165 Heat shock protein beta-1 Human genes 0.000 claims description 22
- MQUFAARYGOUYEV-UWEXFCAOSA-N Quillaic acid Natural products CC1(C)CC[C@@]2([C@H](O)C[C@]3(C)C(=CC[C@H]4[C@@]5(C)CC[C@H](O)[C@](C)(C=O)[C@H]5CC[C@@]34C)[C@H]2C1)C(=O)O MQUFAARYGOUYEV-UWEXFCAOSA-N 0.000 claims description 20
- MQUFAARYGOUYEV-UAWZMHPWSA-N quillaic acid Chemical compound C1C[C@H](O)[C@@](C)(C=O)[C@@H]2CC[C@@]3(C)[C@]4(C)C[C@@H](O)[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MQUFAARYGOUYEV-UAWZMHPWSA-N 0.000 claims description 20
- 102100038837 2-Hydroxyacid oxidase 1 Human genes 0.000 claims description 19
- 102000003855 L-lactate dehydrogenase Human genes 0.000 claims description 19
- 108700023483 L-lactate dehydrogenases Proteins 0.000 claims description 19
- 101710180553 Proprotein convertase subtilisin/kexin type 9 Proteins 0.000 claims description 19
- 108010062584 glycollate oxidase Proteins 0.000 claims description 19
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 19
- PAIBKVQNJKUVCE-UHFFFAOYSA-N Gypsogeninsaeure Natural products C1CC(O)C(C)(C(O)=O)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C PAIBKVQNJKUVCE-UHFFFAOYSA-N 0.000 claims description 17
- 108700011259 MicroRNAs Proteins 0.000 claims description 17
- GCGBHJLBFAPRDB-KCVAUKQGSA-N Scutellaric acid Natural products CC1(C)CC[C@@]2(CC[C@@]3(C)[C@@H]4CC[C@H]5[C@@](C)(CO)[C@H](O)CC[C@]5(C)[C@H]4CC=C3[C@@H]2C1)C(=O)O GCGBHJLBFAPRDB-KCVAUKQGSA-N 0.000 claims description 17
- 108020004999 messenger RNA Proteins 0.000 claims description 17
- 231100000654 protein toxin Toxicity 0.000 claims description 17
- UXFQFBNBSPQBJW-UHFFFAOYSA-N 2-amino-2-methylpropane-1,3-diol Chemical compound OCC(N)(C)CO UXFQFBNBSPQBJW-UHFFFAOYSA-N 0.000 claims description 16
- KZMAWJRXKGLWGS-UHFFFAOYSA-N 2-chloro-n-[4-(4-methoxyphenyl)-1,3-thiazol-2-yl]-n-(3-methoxypropyl)acetamide Chemical compound S1C(N(C(=O)CCl)CCCOC)=NC(C=2C=CC(OC)=CC=2)=C1 KZMAWJRXKGLWGS-UHFFFAOYSA-N 0.000 claims description 16
- 102000004190 Enzymes Human genes 0.000 claims description 16
- 108090000790 Enzymes Proteins 0.000 claims description 16
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 claims description 16
- QMHCWDVPABYZMC-UHFFFAOYSA-N 3-Hydroxy-23-oxo-olean-12-en-28-saeure Natural products C1CC(O)C(C)(C=O)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C QMHCWDVPABYZMC-UHFFFAOYSA-N 0.000 claims description 15
- QMHCWDVPABYZMC-RFVOPWELSA-N gypsogenin Natural products CC1(C)CC[C@@]2(CC[C@]3(C)C(=CC[C@@H]4[C@@]5(C)CC[C@H](O)[C@](C)(C=O)[C@H]5CC[C@@]34C)[C@H]2C1)C(=O)O QMHCWDVPABYZMC-RFVOPWELSA-N 0.000 claims description 15
- 210000004962 mammalian cell Anatomy 0.000 claims description 15
- 239000000074 antisense oligonucleotide Substances 0.000 claims description 14
- 238000012230 antisense oligonucleotides Methods 0.000 claims description 14
- 229940079593 drug Drugs 0.000 claims description 14
- QMHCWDVPABYZMC-MYPRUECHSA-N 3beta-hydroxy-23-oxoolean-12-en-28-oic acid Chemical compound C1C[C@H](O)[C@@](C)(C=O)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C QMHCWDVPABYZMC-MYPRUECHSA-N 0.000 claims description 13
- 239000003085 diluting agent Substances 0.000 claims description 12
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 12
- 238000012546 transfer Methods 0.000 claims description 12
- 108020004414 DNA Proteins 0.000 claims description 11
- 108010084592 Saporins Proteins 0.000 claims description 11
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims description 10
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 10
- 230000009467 reduction Effects 0.000 claims description 9
- 108091033760 Oncomir Proteins 0.000 claims description 8
- 239000001257 hydrogen Substances 0.000 claims description 8
- 229910052739 hydrogen Inorganic materials 0.000 claims description 8
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 8
- 210000005229 liver cell Anatomy 0.000 claims description 8
- 150000002482 oligosaccharides Chemical class 0.000 claims description 8
- 150000003384 small molecules Chemical class 0.000 claims description 8
- IUTPJBLLJJNPAJ-UHFFFAOYSA-N 3-(2,5-dioxopyrrol-1-yl)propanoic acid Chemical compound OC(=O)CCN1C(=O)C=CC1=O IUTPJBLLJJNPAJ-UHFFFAOYSA-N 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 7
- 230000001594 aberrant effect Effects 0.000 claims description 7
- 230000006196 deacetylation Effects 0.000 claims description 7
- 238000003381 deacetylation reaction Methods 0.000 claims description 7
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 claims description 6
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 claims description 6
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 claims description 6
- 108091027967 Small hairpin RNA Proteins 0.000 claims description 6
- IEDXPSOJFSVCKU-HOKPPMCLSA-N [4-[[(2S)-5-(carbamoylamino)-2-[[(2S)-2-[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-3-[[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate Chemical compound CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)c1ccccc1)OC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCc1ccc(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN2C(=O)CCC2=O)C(C)C)cc1)C(C)C IEDXPSOJFSVCKU-HOKPPMCLSA-N 0.000 claims description 6
- 229930191339 dianthin Natural products 0.000 claims description 6
- 150000002772 monosaccharides Chemical class 0.000 claims description 6
- 229940100243 oleanolic acid Drugs 0.000 claims description 6
- 229920001542 oligosaccharide Polymers 0.000 claims description 6
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 claims description 6
- 101710184696 5-aminolevulinate synthase, nonspecific, mitochondrial Proteins 0.000 claims description 5
- 101710095342 Apolipoprotein B Proteins 0.000 claims description 5
- 102100040202 Apolipoprotein B-100 Human genes 0.000 claims description 5
- 239000004019 antithrombin Substances 0.000 claims description 5
- 230000006870 function Effects 0.000 claims description 5
- MFRNYXJJRJQHNW-DEMKXPNLSA-N (2s)-2-[[(2r,3r)-3-methoxy-3-[(2s)-1-[(3r,4s,5s)-3-methoxy-5-methyl-4-[methyl-[(2s)-3-methyl-2-[[(2s)-3-methyl-2-(methylamino)butanoyl]amino]butanoyl]amino]heptanoyl]pyrrolidin-2-yl]-2-methylpropanoyl]amino]-3-phenylpropanoic acid Chemical compound CN[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MFRNYXJJRJQHNW-DEMKXPNLSA-N 0.000 claims description 4
- OTLLEIBWKHEHGU-UHFFFAOYSA-N 2-[5-[[5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy]-3,4-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-4-phosphonooxyhexanedioic acid Chemical compound C1=NC=2C(N)=NC=NC=2N1C(C(C1O)O)OC1COC1C(CO)OC(OC(C(O)C(OP(O)(O)=O)C(O)C(O)=O)C(O)=O)C(O)C1O OTLLEIBWKHEHGU-UHFFFAOYSA-N 0.000 claims description 4
- 108010066676 Abrin Proteins 0.000 claims description 4
- AXNVHPCVMSNXNP-GKTCLTPXSA-N Aescin Natural products O=C(O[C@H]1[C@@H](OC(=O)C)[C@]2(CO)[C@@H](O)C[C@@]3(C)[C@@]4(C)[C@@H]([C@]5(C)[C@H]([C@](CO)(C)[C@@H](O[C@@H]6[C@@H](O[C@H]7[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O7)[C@@H](O)[C@H](O[C@H]7[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O7)[C@@H](C(=O)O)O6)CC5)CC4)CC=C3[C@@H]2CC1(C)C)/C(=C/C)/C AXNVHPCVMSNXNP-GKTCLTPXSA-N 0.000 claims description 4
- 231100000699 Bacterial toxin Toxicity 0.000 claims description 4
- 102000016607 Diphtheria Toxin Human genes 0.000 claims description 4
- 108010053187 Diphtheria Toxin Proteins 0.000 claims description 4
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 4
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 claims description 4
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 claims description 4
- 231100000742 Plant toxin Toxicity 0.000 claims description 4
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims description 4
- 108090000829 Ribosome Inactivating Proteins Proteins 0.000 claims description 4
- 108010039491 Ricin Proteins 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- 239000000688 bacterial toxin Substances 0.000 claims description 4
- 229960003668 docetaxel Drugs 0.000 claims description 4
- VQNATVDKACXKTF-XELLLNAOSA-N duocarmycin Chemical compound COC1=C(OC)C(OC)=C2NC(C(=O)N3C4=CC(=O)C5=C([C@@]64C[C@@H]6C3)C=C(N5)C(=O)OC)=CC2=C1 VQNATVDKACXKTF-XELLLNAOSA-N 0.000 claims description 4
- ZVYVPGLRVWUPMP-FYSMJZIKSA-N exatecan Chemical compound C1C[C@H](N)C2=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC3=CC(F)=C(C)C1=C32 ZVYVPGLRVWUPMP-FYSMJZIKSA-N 0.000 claims description 4
- 229950009429 exatecan Drugs 0.000 claims description 4
- 239000002095 exotoxin Substances 0.000 claims description 4
- 231100000776 exotoxin Toxicity 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- 229960005558 mertansine Drugs 0.000 claims description 4
- 108010022050 mistletoe lectin I Proteins 0.000 claims description 4
- 108010010621 modeccin Proteins 0.000 claims description 4
- 231100000590 oncogenic Toxicity 0.000 claims description 4
- 230000002246 oncogenic effect Effects 0.000 claims description 4
- 239000003123 plant toxin Substances 0.000 claims description 4
- YUOCYTRGANSSRY-UHFFFAOYSA-N pyrrolo[2,3-i][1,2]benzodiazepine Chemical compound C1=CN=NC2=C3C=CN=C3C=CC2=C1 YUOCYTRGANSSRY-UHFFFAOYSA-N 0.000 claims description 4
- QLPRYZXNWYTFCI-UHFFFAOYSA-N saikosaponin D Natural products CC1OC(OC2CCC3(C)C(CCC4(C)C3C=CC56OCC7(CCC(C)(C)CC57)C(O)CC46C)C2(C)CO)C(O)C(O)C1OC8OC(CO)C(O)C(O)C8O QLPRYZXNWYTFCI-UHFFFAOYSA-N 0.000 claims description 4
- PQPVAGWUNWFCJE-UHFFFAOYSA-N saikosaponin a Natural products CC1OC(OC2CCC3(C)C(C2)C(C)(CO)CC4(C)C3C=CC56OCC7(CCC(C)(C)CC57)C(O)CC46C)C(O)C(OC8OC(CO)C(O)C(O)C8O)C1O PQPVAGWUNWFCJE-UHFFFAOYSA-N 0.000 claims description 4
- 230000001629 suppression Effects 0.000 claims description 4
- 229930182493 triterpene saponin Natural products 0.000 claims description 4
- 230000003612 virological effect Effects 0.000 claims description 4
- 108020004491 Antisense DNA Proteins 0.000 claims description 3
- 241000282412 Homo Species 0.000 claims description 3
- 230000003042 antagnostic effect Effects 0.000 claims description 3
- 239000003816 antisense DNA Substances 0.000 claims description 3
- 208000035475 disorder Diseases 0.000 claims description 3
- 239000012634 fragment Substances 0.000 claims description 3
- PIGTXFOGKFOFTO-FVFWYJKVSA-N (2S,3S,4S,5R,6R)-6-[[(3S,4S,4aR,6aR,6bS,8R,8aR,12aS,14aR,14bR)-8a-carboxy-4-formyl-8-hydroxy-4,6a,6b,11,11,14b-hexamethyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound O([C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(C=O)C)C)(C)C[C@@H](O)[C@]1(CCC(C[C@H]14)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O PIGTXFOGKFOFTO-FVFWYJKVSA-N 0.000 claims description 2
- CYJWWQALTIKOAG-FLORRLIPSA-N (2S,4aR,6aR,6aS,6bR,8aR,9R,10R,11S,12aR,14bS)-10,11-dihydroxy-9-(hydroxymethyl)-2-methoxycarbonyl-2,6a,6b,9,12a-pentamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylic acid Chemical compound C1[C@H](O)[C@H](O)[C@@](C)(CO)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@](C(=O)OC)(C)C[C@H]5C4=CC[C@@H]3[C@]21C CYJWWQALTIKOAG-FLORRLIPSA-N 0.000 claims description 2
- ORFNVPGICPYLJV-YTVPMEHESA-N (2s)-2-[[(2r,3r)-3-[(2s)-1-[(3r,4s,5s)-4-[[(2s)-2-[[(2s)-2-[6-(2,5-dioxopyrrol-1-yl)hexanoyl-methylamino]-3-methylbutanoyl]amino]-3-methylbutanoyl]-methylamino]-3-methoxy-5-methylheptanoyl]pyrrolidin-2-yl]-3-methoxy-2-methylpropanoyl]amino]-3-phenylpropan Chemical compound C([C@H](NC(=O)[C@H](C)[C@@H](OC)[C@@H]1CCCN1C(=O)C[C@H]([C@H]([C@@H](C)CC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)CCCCCN1C(C=CC1=O)=O)C(C)C)OC)C(O)=O)C1=CC=CC=C1 ORFNVPGICPYLJV-YTVPMEHESA-N 0.000 claims description 2
- YFESOSRPNPYODN-RSMWSHJLSA-N (2s,3s,4s,5r,6r)-6-[[(4s,6ar,6bs,8r,8ar,9r,10r,14br)-9-acetyloxy-8-hydroxy-4,8a-bis(hydroxymethyl)-4,6a,6b,11,11,14b-hexamethyl-10-[(z)-2-methylbut-2-enoyl]oxy-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-4-hydroxy-3,5-bis[[(2s,3r,4s, Chemical compound O([C@@H]1[C@H](O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)OC1CC[C@]2(C)C3CC=C4[C@@]([C@@]3(CCC2[C@]1(CO)C)C)(C)C[C@@H](O)[C@@]1(CO)[C@@H](OC(C)=O)[C@@H](C(CC14)(C)C)OC(=O)C(\C)=C/C)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O.O([C@@H]1[C@H](O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)OC1CC[C@]2(C)C3CC=C4[C@@]([C@@]3(CCC2[C@]1(CO)C)C)(C)C[C@@H](O)[C@@]1(CO)[C@@H](OC(C)=O)[C@@H](C(CC14)(C)C)OC(=O)C(/C)=C/C)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YFESOSRPNPYODN-RSMWSHJLSA-N 0.000 claims description 2
- DLKUYSQUHXBYPB-NSSHGSRYSA-N (2s,4r)-4-[[2-[(1r,3r)-1-acetyloxy-4-methyl-3-[3-methylbutanoyloxymethyl-[(2s,3s)-3-methyl-2-[[(2r)-1-methylpiperidine-2-carbonyl]amino]pentanoyl]amino]pentyl]-1,3-thiazole-4-carbonyl]amino]-2-methyl-5-(4-methylphenyl)pentanoic acid Chemical compound N([C@@H]([C@@H](C)CC)C(=O)N(COC(=O)CC(C)C)[C@H](C[C@@H](OC(C)=O)C=1SC=C(N=1)C(=O)N[C@H](C[C@H](C)C(O)=O)CC=1C=CC(C)=CC=1)C(C)C)C(=O)[C@H]1CCCCN1C DLKUYSQUHXBYPB-NSSHGSRYSA-N 0.000 claims description 2
- NTWLPZMPTFQYQI-UHFFFAOYSA-N (3alpha)-olean-12-ene-3,23-diol Natural products C1CC(O)C(C)(CO)C2CCC3(C)C4(C)CCC5(C)CCC(C)(C)CC5C4=CCC3C21C NTWLPZMPTFQYQI-UHFFFAOYSA-N 0.000 claims description 2
- SVUOLADPCWQTTE-UHFFFAOYSA-N 1h-1,2-benzodiazepine Chemical compound N1N=CC=CC2=CC=CC=C12 SVUOLADPCWQTTE-UHFFFAOYSA-N 0.000 claims description 2
- FBUTXZSKZCQABC-UHFFFAOYSA-N 2-amino-1-methyl-7h-purine-6-thione Chemical compound S=C1N(C)C(N)=NC2=C1NC=N2 FBUTXZSKZCQABC-UHFFFAOYSA-N 0.000 claims description 2
- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 claims description 2
- FJHBVJOVLFPMQE-QFIPXVFZSA-N 7-Ethyl-10-Hydroxy-Camptothecin Chemical compound C1=C(O)C=C2C(CC)=C(CN3C(C4=C([C@@](C(=O)OC4)(O)CC)C=C33)=O)C3=NC2=C1 FJHBVJOVLFPMQE-QFIPXVFZSA-N 0.000 claims description 2
- AXNVHPCVMSNXNP-OXPBSUTMSA-N Aescin Chemical compound O([C@@H]1[C@H](O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@]1(CO)C)C)(C)C[C@@H](O)[C@@]1(CO)[C@@H](OC(C)=O)[C@@H](C(C[C@H]14)(C)C)OC(=O)C(\C)=C/C)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O AXNVHPCVMSNXNP-OXPBSUTMSA-N 0.000 claims description 2
- 108010027164 Amanitins Proteins 0.000 claims description 2
- 102100022987 Angiogenin Human genes 0.000 claims description 2
- 101710094856 Apoptin Proteins 0.000 claims description 2
- 108091023037 Aptamer Proteins 0.000 claims description 2
- BJFNIGZQPQQAFL-UTPDYZQNSA-N Assamsaponin F Chemical compound O([C@@H]1[C@@H](O)[C@@H](O)CO[C@H]1O[C@H]1[C@H](O)[C@H](O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O1)O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(C=O)C)C)(C)C[C@H]([C@@]1(CO)[C@@H](OC(C)=O)[C@@H](C(C[C@H]14)(C)C)OC(=O)C(\C)=C/C)OC(C)=O)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O BJFNIGZQPQQAFL-UTPDYZQNSA-N 0.000 claims description 2
- 102000009016 Cholera Toxin Human genes 0.000 claims description 2
- 108010049048 Cholera Toxin Proteins 0.000 claims description 2
- 108010051219 Cre recombinase Proteins 0.000 claims description 2
- 229930188224 Cryptophycin Natural products 0.000 claims description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 claims description 2
- COVOPPXLDJVUSC-UHFFFAOYSA-N Digitogenin Natural products CC1C(C2(CCC3C4(C)CC(O)C(O)CC4CCC3C2C2O)C)C2OC11CCC(C)CO1 COVOPPXLDJVUSC-UHFFFAOYSA-N 0.000 claims description 2
- QRLVDLBMBULFAL-UHFFFAOYSA-N Digitonin Natural products CC1CCC2(OC1)OC3C(O)C4C5CCC6CC(OC7OC(CO)C(OC8OC(CO)C(O)C(OC9OCC(O)C(O)C9OC%10OC(CO)C(O)C(OC%11OC(CO)C(O)C(O)C%11O)C%10O)C8O)C(O)C7O)C(O)CC6(C)C5CCC4(C)C3C2C QRLVDLBMBULFAL-UHFFFAOYSA-N 0.000 claims description 2
- 101710082714 Exotoxin A Proteins 0.000 claims description 2
- 102000001398 Granzyme Human genes 0.000 claims description 2
- 108060005986 Granzyme Proteins 0.000 claims description 2
- LAHSXXNOJMWHBH-WVPBMNGESA-N Gypsoside Natural products O=C(O)[C@H]1[C@H](O[C@H]2[C@@H](O)[C@H](O)[C@H](O[C@@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)[C@H](CO)O2)[C@H](O[C@H]2[C@@H](O)[C@@H](O)[C@@H](O)CO2)[C@@H](O)[C@@H](O[C@@H]2[C@](C=O)(C)[C@@H]3[C@](C)([C@H]4[C@](C)([C@]5(C)C([C@H]6[C@](C(=O)O[C@H]7[C@H](O[C@@H]8[C@H](O[C@@H]9[C@H](O)[C@H](O)[C@H](O)CO9)[C@H](O)[C@H](O)CO8)[C@@H](O)[C@@H](O[C@@H]8[C@@H](O)[C@@H](O[C@H]9[C@@H](O)[C@@H](O)[C@@H](O)CO9)[C@H](O)[C@@H](C)O8)[C@@H](C)O7)(CC5)CCC(C)(C)C6)=CC4)CC3)CC2)O1 LAHSXXNOJMWHBH-WVPBMNGESA-N 0.000 claims description 2
- GCGBHJLBFAPRDB-UHFFFAOYSA-N Hederagenin Natural products CC1(C)CCC2(CCC3(C)C4CCC5C(C)(CO)C(O)CCC5(C)C4CC=C3C2C1)C(=O)O GCGBHJLBFAPRDB-UHFFFAOYSA-N 0.000 claims description 2
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 claims description 2
- 229930012538 Paclitaxel Natural products 0.000 claims description 2
- 102000002508 Peptide Elongation Factors Human genes 0.000 claims description 2
- 108010068204 Peptide Elongation Factors Proteins 0.000 claims description 2
- ZMXKPCHQLHYTHY-UHFFFAOYSA-N Phytolaccasaponin E Natural products C12CC(C(=O)OC)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(CO)C)C)(C)C1=CCC2C3(C)CC(O)C4OC(C(C1O)O)OCC1OC1OC(CO)C(O)C(O)C1O ZMXKPCHQLHYTHY-UHFFFAOYSA-N 0.000 claims description 2
- YRHWKFMGEDDGIJ-UHFFFAOYSA-N Phytolaccoside E Chemical compound C12CC(C(=O)OC)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(CO)C)C)(C)C1=CCC2C3(C)CC(O)C4OC(C(C1O)O)COC1OC1OC(CO)C(O)C(O)C1O YRHWKFMGEDDGIJ-UHFFFAOYSA-N 0.000 claims description 2
- 241000245063 Primula Species 0.000 claims description 2
- 235000000497 Primula Nutrition 0.000 claims description 2
- 241001092473 Quillaja Species 0.000 claims description 2
- 235000009001 Quillaja saponaria Nutrition 0.000 claims description 2
- OWPCHSCAPHNHAV-UHFFFAOYSA-N Rhizoxin Natural products C1C(O)C2(C)OC2C=CC(C)C(OC(=O)C2)CC2CC2OC2C(=O)OC1C(C)C(OC)C(C)=CC=CC(C)=CC1=COC(C)=N1 OWPCHSCAPHNHAV-UHFFFAOYSA-N 0.000 claims description 2
- 108010083644 Ribonucleases Proteins 0.000 claims description 2
- 102000006382 Ribonucleases Human genes 0.000 claims description 2
- KYWSCMDFVARMPN-MSSMMRRTSA-N Saikosaponin A Chemical compound O([C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@]([C@H]3[C@]([C@@H]4[C@@]([C@@]5(C[C@H](O)[C@]67CO[C@]5([C@@H]6CC(C)(C)CC7)C=C4)C)(C)CC3)(C)CC2)(C)CO)O[C@@H]([C@@H]1O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O KYWSCMDFVARMPN-MSSMMRRTSA-N 0.000 claims description 2
- KYWSCMDFVARMPN-LCSVLAELSA-N Saikosaponin D Chemical compound O([C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@]([C@H]3[C@]([C@@H]4[C@@]([C@@]5(C[C@@H](O)[C@]67CO[C@]5([C@@H]6CC(C)(C)CC7)C=C4)C)(C)CC3)(C)CC2)(C)CO)O[C@@H]([C@@H]1O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O KYWSCMDFVARMPN-LCSVLAELSA-N 0.000 claims description 2
- KEOITPILCOILGM-FCWYPSSHSA-N Sapindoside A Natural products O=C(O)[C@]12[C@H](C=3[C@](C)([C@@]4(C)[C@@H]([C@]5(C)[C@H]([C@@](CO)(C)[C@@H](O[C@@H]6[C@@H](O[C@H]7[C@H](O)[C@@H](O)[C@@H](O)[C@H](C)O7)[C@H](O)[C@H](O)CO6)CC5)CC4)CC=3)CC1)CC(C)(C)CC2 KEOITPILCOILGM-FCWYPSSHSA-N 0.000 claims description 2
- 108010079723 Shiga Toxin Proteins 0.000 claims description 2
- 101710084578 Short neurotoxin 1 Proteins 0.000 claims description 2
- 101710182532 Toxin a Proteins 0.000 claims description 2
- 108090000704 Tubulin Proteins 0.000 claims description 2
- 102000004243 Tubulin Human genes 0.000 claims description 2
- 108010061323 Type 2 Ribosome Inactivating Proteins Proteins 0.000 claims description 2
- 108010046334 Urease Proteins 0.000 claims description 2
- 101800004982 Volkensin A chain Proteins 0.000 claims description 2
- VUEGHZSQVJADCO-UGZFTLGKSA-N [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl] (4aS,6aR,6aS,6bR,8aR,9R,10S,12aR,14bS)-10-[(2S,3R,4S,5S)-3-[(2S,3R,4R,5S,6S)-3,5-dihydroxy-6-methyl-4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-4,5-dihydroxyoxan-2-yl]oxy-9-(hydroxymethyl)-2,2,6a,6b,9,12a-hexamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylate Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)[C@]12CCC(C)(C)C[C@H]1C1=CC[C@H]3[C@@]([C@@]1(CC2)C)(C)CC[C@@H]1[C@]3(C)CC[C@@H]([C@@]1(C)CO)O[C@@H]1OC[C@H](O)[C@H](O)[C@H]1O[C@@H]1O[C@H]([C@@H]([C@@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H]1O)O)C)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VUEGHZSQVJADCO-UGZFTLGKSA-N 0.000 claims description 2
- UZQJVUCHXGYFLQ-AYDHOLPZSA-N [(2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-4-[(2r,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-6-(hy Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(C=O)C)C)(C)CC(O)[C@]1(CCC(CC14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O[C@H]4[C@@H]([C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)[C@H](O)[C@@H](CO)O4)O)[C@H](O)[C@@H](CO)O3)O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UZQJVUCHXGYFLQ-AYDHOLPZSA-N 0.000 claims description 2
- GFPLPBCJRRNZHM-ICUGHKHQSA-N [(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl] (4as,6ar,6as,6br,8ar,9r,10s,12ar,14bs)-10-[(2s,3r,4s,5s)-4,5-dihydroxy-3-[(2s,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan- Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](O[C@@H]2[C@@]([C@H]3[C@]([C@@H]4[C@@]([C@@]5(CC[C@]6(CCC(C)(C)C[C@H]6C5=CC4)C(=O)O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)O4)O)C)(C)CC3)(C)CC2)(C)CO)OC[C@H](O)[C@@H]1O GFPLPBCJRRNZHM-ICUGHKHQSA-N 0.000 claims description 2
- HKGATZAPXCCEJR-OWRSNIELSA-N [4-[[(2s)-2-[[(2s)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoylamino]-3-methylbutanoyl]amino]propanoyl]amino]phenyl]methyl (6s,6as)-3-[5-[[(6as)-2-methoxy-8-methyl-1 Chemical compound N([C@H](C(=O)N[C@@H](C)C(=O)NC1=CC=C(C=C1)COC(=O)N1C=2C=C(C(=CC=2C(=O)N2C=C(C)C[C@H]2[C@@H]1O)OC)OCCCCCOC1=CC2=C(C(N3C=C(C)C[C@H]3C=N2)=O)C=C1OC)C(C)C)C(=O)CCOCCOCCOCCOCCOCCOCCOCCOCCNC(=O)CCN1C(=O)C=CC1=O HKGATZAPXCCEJR-OWRSNIELSA-N 0.000 claims description 2
- KBYYTUYPCGPQNK-UHFFFAOYSA-N alpha-hederin Natural products CC1OC(OC2C(O)C(CO)OC2OC3CCC4(C)C(CCC5(C)C4CC=C6C7CC(C)(C)CCC7(CCC56C)C(=O)O)C3(C)CO)C(O)C(O)C1O KBYYTUYPCGPQNK-UHFFFAOYSA-N 0.000 claims description 2
- 108010001818 alpha-sarcin Proteins 0.000 claims description 2
- 101150078331 ama-1 gene Proteins 0.000 claims description 2
- CIORWBWIBBPXCG-JZTFPUPKSA-N amanitin Chemical compound O=C1N[C@@H](CC(N)=O)C(=O)N2CC(O)C[C@H]2C(=O)N[C@@H](C(C)[C@@H](O)CO)C(=O)N[C@@H](C2)C(=O)NCC(=O)N[C@@H](C(C)CC)C(=O)NCC(=O)N[C@H]1CS(=O)C1=C2C2=CC=C(O)C=C2N1 CIORWBWIBBPXCG-JZTFPUPKSA-N 0.000 claims description 2
- 108010072788 angiogenin Proteins 0.000 claims description 2
- 229940045799 anthracyclines and related substance Drugs 0.000 claims description 2
- VUEGHZSQVJADCO-UHFFFAOYSA-N aralia saponin 3 Natural products OC1C(OC2C(C(O)C(O)C(CO)O2)O)C(O)C(C)OC1OC1C(O)C(O)COC1OC(C1(C)CO)CCC2(C)C1CCC(C1(CC3)C)(C)C2CC=C1C1CC(C)(C)CCC13C(=O)OC(C(C(O)C1O)O)OC1COC1OC(CO)C(O)C(O)C1O VUEGHZSQVJADCO-UHFFFAOYSA-N 0.000 claims description 2
- LCGQSISMCSVPJV-UHFFFAOYSA-N assamsaponin F Natural products CC=C(C)/C(=O)OC1C(OC(=O)C)C2(CO)C(CC3(C)C(=CCC4C5(C)CCC(OC6OC(C(O)C(OC7OC(O)C(O)CC7OC8OC(CO)C(O)C(O)C8O)C6OC9OC(CO)C(O)C(O)C9O)C(=O)O)C(C)(C=O)C5CCC34C)C2CC1(C)C)OC(=O)C LCGQSISMCSVPJV-UHFFFAOYSA-N 0.000 claims description 2
- 229940049706 benzodiazepine Drugs 0.000 claims description 2
- HXCHCVDVKSCDHU-LULTVBGHSA-N calicheamicin Chemical compound C1[C@H](OC)[C@@H](NCC)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@](C/3=C/CSSSC)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HXCHCVDVKSCDHU-LULTVBGHSA-N 0.000 claims description 2
- 229930195731 calicheamicin Natural products 0.000 claims description 2
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical class C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 claims description 2
- 229960004316 cisplatin Drugs 0.000 claims description 2
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 2
- MDZKJHQSJHYOHJ-UHFFFAOYSA-N crataegolic acid Natural products C1C(O)C(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MDZKJHQSJHYOHJ-UHFFFAOYSA-N 0.000 claims description 2
- 108010006226 cryptophycin Proteins 0.000 claims description 2
- PSNOPSMXOBPNNV-VVCTWANISA-N cryptophycin 1 Chemical compound C1=C(Cl)C(OC)=CC=C1C[C@@H]1C(=O)NC[C@@H](C)C(=O)O[C@@H](CC(C)C)C(=O)O[C@H]([C@H](C)[C@@H]2[C@H](O2)C=2C=CC=CC=2)C/C=C/C(=O)N1 PSNOPSMXOBPNNV-VVCTWANISA-N 0.000 claims description 2
- PSNOPSMXOBPNNV-UHFFFAOYSA-N cryptophycin-327 Natural products C1=C(Cl)C(OC)=CC=C1CC1C(=O)NCC(C)C(=O)OC(CC(C)C)C(=O)OC(C(C)C2C(O2)C=2C=CC=CC=2)CC=CC(=O)N1 PSNOPSMXOBPNNV-UHFFFAOYSA-N 0.000 claims description 2
- 229960004397 cyclophosphamide Drugs 0.000 claims description 2
- COVOPPXLDJVUSC-JPYPKGSXSA-N digitogenin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)C[C@@H](O)[C@H](O)C[C@@H]4CC[C@H]3[C@@H]2[C@@H]1O)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 COVOPPXLDJVUSC-JPYPKGSXSA-N 0.000 claims description 2
- UVYVLBIGDKGWPX-KUAJCENISA-N digitonin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)C[C@@H](O)[C@H](O[C@H]5[C@@H]([C@@H](O)[C@@H](O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)CO7)O)[C@H](O)[C@@H](CO)O6)O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O7)O)[C@@H](O)[C@@H](CO)O6)O)[C@@H](CO)O5)O)C[C@@H]4CC[C@H]3[C@@H]2[C@@H]1O)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 UVYVLBIGDKGWPX-KUAJCENISA-N 0.000 claims description 2
- UVYVLBIGDKGWPX-UHFFFAOYSA-N digitonine Natural products CC1C(C2(CCC3C4(C)CC(O)C(OC5C(C(O)C(OC6C(C(OC7C(C(O)C(O)CO7)O)C(O)C(CO)O6)OC6C(C(OC7C(C(O)C(O)C(CO)O7)O)C(O)C(CO)O6)O)C(CO)O5)O)CC4CCC3C2C2O)C)C2OC11CCC(C)CO1 UVYVLBIGDKGWPX-UHFFFAOYSA-N 0.000 claims description 2
- GFPLPBCJRRNZHM-UHFFFAOYSA-N dipsacoside B Natural products CC1OC(OC2C(O)C(O)COC2OC2CCC3(C)C(CCC4(C)C3CC=C3C5CC(C)(C)CCC5(CCC43C)C(=O)OC3OC(COC4OC(CO)C(O)C(O)C4O)C(O)C(O)C3O)C2(C)CO)C(O)C(O)C1O GFPLPBCJRRNZHM-UHFFFAOYSA-N 0.000 claims description 2
- 229960004679 doxorubicin Drugs 0.000 claims description 2
- 229960005501 duocarmycin Drugs 0.000 claims description 2
- 229930184221 duocarmycin Natural products 0.000 claims description 2
- 229960005420 etoposide Drugs 0.000 claims description 2
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 claims description 2
- PAIBKVQNJKUVCE-JUENUIDLSA-N gypsogenic acid Chemical compound C1C[C@H](O)[C@@](C)(C(O)=O)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C PAIBKVQNJKUVCE-JUENUIDLSA-N 0.000 claims description 2
- PAIBKVQNJKUVCE-HDHBZPKHSA-N gypsogenic acid Natural products CC1(C)CC[C@@]2(CC[C@]3(C)C(=CC[C@@H]4[C@@]5(C)CC[C@H](O)[C@@](C)([C@H]5CC[C@@]34C)C(O)=O)[C@H]2C1)C(O)=O PAIBKVQNJKUVCE-HDHBZPKHSA-N 0.000 claims description 2
- PGOYMURMZNDHNS-MYPRUECHSA-N hederagenin Chemical compound C1C[C@H](O)[C@@](C)(CO)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C PGOYMURMZNDHNS-MYPRUECHSA-N 0.000 claims description 2
- KEOITPILCOILGM-LLJOFIFVSA-N kalopanaxsaponin A Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](O[C@@H]2[C@@]([C@H]3[C@]([C@@H]4[C@@]([C@@]5(CC[C@]6(CCC(C)(C)C[C@H]6C5=CC4)C(O)=O)C)(C)CC3)(C)CC2)(C)CO)OC[C@H](O)[C@@H]1O KEOITPILCOILGM-LLJOFIFVSA-N 0.000 claims description 2
- 229960000485 methotrexate Drugs 0.000 claims description 2
- 229960001156 mitoxantrone Drugs 0.000 claims description 2
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 claims description 2
- 108010093470 monomethyl auristatin E Proteins 0.000 claims description 2
- 108010059074 monomethylauristatin F Proteins 0.000 claims description 2
- 239000002636 mycotoxin Substances 0.000 claims description 2
- 229950007318 ozogamicin Drugs 0.000 claims description 2
- 229960001592 paclitaxel Drugs 0.000 claims description 2
- CYJWWQALTIKOAG-UHFFFAOYSA-N phytolaccagenin Natural products C1C(O)C(O)C(C)(CO)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C(=O)OC)(C)CC5C4=CCC3C21C CYJWWQALTIKOAG-UHFFFAOYSA-N 0.000 claims description 2
- 108700028325 pokeweed antiviral Proteins 0.000 claims description 2
- OWPCHSCAPHNHAV-LMONGJCWSA-N rhizoxin Chemical compound C/C([C@H](OC)[C@@H](C)[C@@H]1C[C@H](O)[C@]2(C)O[C@@H]2/C=C/[C@@H](C)[C@]2([H])OC(=O)C[C@@](C2)(C[C@@H]2O[C@H]2C(=O)O1)[H])=C\C=C\C(\C)=C\C1=COC(C)=N1 OWPCHSCAPHNHAV-LMONGJCWSA-N 0.000 claims description 2
- HNMATTJJEPZZMM-BPKVFSPJSA-N s-[(2r,3s,4s,6s)-6-[[(2r,3s,4s,5r,6r)-5-[(2s,4s,5s)-5-[acetyl(ethyl)amino]-4-methoxyoxan-2-yl]oxy-6-[[(2s,5z,9r,13e)-13-[2-[[4-[(2e)-2-[1-[4-(4-amino-4-oxobutoxy)phenyl]ethylidene]hydrazinyl]-2-methyl-4-oxobutan-2-yl]disulfanyl]ethylidene]-9-hydroxy-12-(m Chemical compound C1[C@H](OC)[C@@H](N(CC)C(C)=O)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@@](C/3=C/CSSC(C)(C)CC(=O)N\N=C(/C)C=3C=CC(OCCCC(N)=O)=CC=3)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HNMATTJJEPZZMM-BPKVFSPJSA-N 0.000 claims description 2
- XNZLMZRIGGHITK-VKXBZTRUSA-N soravtansine Chemical compound CO[C@@H]([C@@]1(O)C[C@H](OC(=O)N1)[C@@H](C)[C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(=O)CCC(C)(C)SSCC[C@@H](C(O)=O)S(O)(=O)=O)CC(=O)N1C)\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2.CO[C@@H]([C@@]1(O)C[C@H](OC(=O)N1)[C@@H](C)[C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(=O)CCC(C)(C)SSCC[C@H](C(O)=O)S(O)(=O)=O)CC(=O)N1C)\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 XNZLMZRIGGHITK-VKXBZTRUSA-N 0.000 claims description 2
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims description 2
- NEEQJTVMZAIFID-OBTMRSOISA-N teaseedsaponin I Natural products CC=C(C)C(=O)O[C@H]1[C@H](OC(=O)C(=CC)C)[C@]2(CO)[C@H](O)C[C@]3(C)C(=CC[C@@H]4[C@@]5(C)CC[C@H](O[C@@H]6O[C@@H]([C@@H](O)[C@H](O[C@@H]7OC[C@H](O)[C@H](O)[C@H]7O[C@@H]8O[C@H](CO)[C@@H](O)[C@H](O)[C@H]8O)[C@H]6O[C@@H]9O[C@H](CO)[C@H](O)[C@H](O)[C@H]9O)C(=O)O)[C@@](C)(C=O)[C@@H]5CC[C@@]34C)[C@@H]2CC1(C)C NEEQJTVMZAIFID-OBTMRSOISA-N 0.000 claims description 2
- QBNQXNXXIWAIMM-VMHSQIKKSA-N teaseedsaponin J Natural products CC=C(C)/C(=O)O[C@H]1[C@H](OC(=O)C(=C/C)C)[C@]2(CO)[C@H](O)C[C@]3(C)C(=CC[C@@H]4[C@@]5(C)CC[C@H](O[C@@H]6O[C@@H]([C@@H](O)[C@H](O[C@@H]7OC[C@H](O)[C@H](O)[C@H]7O[C@@H]8OC[C@@H](O)[C@H](O)[C@H]8O)[C@H]6O[C@@H]9O[C@H](CO)[C@H](O)[C@H](O)[C@H]9O)C(=O)O)[C@@](C)(C=O)[C@@H]5CC[C@@]34C)[C@@H]2CC1(C)C QBNQXNXXIWAIMM-VMHSQIKKSA-N 0.000 claims description 2
- UIERETOOQGIECD-ONEGZZNKSA-N tiglic acid Chemical compound C\C=C(/C)C(O)=O UIERETOOQGIECD-ONEGZZNKSA-N 0.000 claims description 2
- 229930184737 tubulysin Natural products 0.000 claims description 2
- 108010043904 volkensin Proteins 0.000 claims description 2
- KQNNSYZQMSOOQH-GLDAUDTLSA-N volkensin Chemical compound C=1([C@@H]2C[C@@H]3O[C@@H](O)C[C@@H]4[C@]5(C)[C@H]6[C@H]([C@H]([C@@]4(C)C3=C2C)O)OC[C@]6(C)[C@H](OC(C)=O)C[C@@H]5OC(=O)C(/C)=C/C)C=COC=1 KQNNSYZQMSOOQH-GLDAUDTLSA-N 0.000 claims description 2
- 230000001419 dependent effect Effects 0.000 claims 2
- AEMOLEFTQBMNLQ-UHFFFAOYSA-N beta-D-galactopyranuronic acid Natural products OC1OC(C(O)=O)C(O)C(O)C1O AEMOLEFTQBMNLQ-UHFFFAOYSA-N 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract description 32
- 101710163968 Antistasin Proteins 0.000 abstract 1
- 102100033775 Collagen alpha-5(IV) chain Human genes 0.000 abstract 1
- 101001098868 Homo sapiens Proprotein convertase subtilisin/kexin type 9 Proteins 0.000 abstract 1
- 235000017709 saponins Nutrition 0.000 description 117
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 96
- 239000000562 conjugate Substances 0.000 description 95
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 84
- 239000011541 reaction mixture Substances 0.000 description 83
- 239000000243 solution Substances 0.000 description 75
- 150000001875 compounds Chemical class 0.000 description 52
- 125000005647 linker group Chemical group 0.000 description 50
- 239000000047 product Substances 0.000 description 48
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 description 36
- 229910000013 Ammonium bicarbonate Inorganic materials 0.000 description 36
- 235000012538 ammonium bicarbonate Nutrition 0.000 description 36
- 239000001099 ammonium carbonate Substances 0.000 description 36
- 210000003494 hepatocyte Anatomy 0.000 description 32
- 235000018102 proteins Nutrition 0.000 description 32
- 239000007787 solid Substances 0.000 description 32
- 102000018616 Apolipoproteins B Human genes 0.000 description 31
- 108010027006 Apolipoproteins B Proteins 0.000 description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 30
- 238000003786 synthesis reaction Methods 0.000 description 29
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 27
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 27
- 102100026144 Transferrin receptor protein 1 Human genes 0.000 description 20
- 238000012226 gene silencing method Methods 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 20
- 101000835093 Homo sapiens Transferrin receptor protein 1 Proteins 0.000 description 19
- 239000002243 precursor Substances 0.000 description 19
- 238000005859 coupling reaction Methods 0.000 description 17
- 239000003480 eluent Substances 0.000 description 16
- QWENRTYMTSOGBR-UHFFFAOYSA-N 1H-1,2,3-Triazole Chemical compound C=1C=NNN=1 QWENRTYMTSOGBR-UHFFFAOYSA-N 0.000 description 15
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 description 14
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 14
- 230000009368 gene silencing by RNA Effects 0.000 description 14
- 125000000539 amino acid group Chemical group 0.000 description 12
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 12
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 11
- 239000000611 antibody drug conjugate Substances 0.000 description 11
- 229940049595 antibody-drug conjugate Drugs 0.000 description 11
- 230000000694 effects Effects 0.000 description 11
- 210000001163 endosome Anatomy 0.000 description 11
- 238000000105 evaporative light scattering detection Methods 0.000 description 11
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 10
- 230000004071 biological effect Effects 0.000 description 10
- 230000021615 conjugation Effects 0.000 description 10
- 238000010195 expression analysis Methods 0.000 description 10
- 239000003643 water by type Substances 0.000 description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 9
- 230000003834 intracellular effect Effects 0.000 description 9
- 125000003729 nucleotide group Chemical group 0.000 description 9
- 238000003570 cell viability assay Methods 0.000 description 8
- 230000008878 coupling Effects 0.000 description 8
- 238000010168 coupling process Methods 0.000 description 8
- 150000007857 hydrazones Chemical class 0.000 description 8
- 239000002773 nucleotide Substances 0.000 description 8
- 230000003389 potentiating effect Effects 0.000 description 8
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 8
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 7
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 7
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 7
- 238000004113 cell culture Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- 230000002708 enhancing effect Effects 0.000 description 7
- 239000012091 fetal bovine serum Substances 0.000 description 7
- 238000006722 reduction reaction Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 238000011529 RT qPCR Methods 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 6
- 235000019253 formic acid Nutrition 0.000 description 6
- 230000037361 pathway Effects 0.000 description 6
- 231100000419 toxicity Toxicity 0.000 description 6
- 230000001988 toxicity Effects 0.000 description 6
- AUDYZXNUHIIGRB-UHFFFAOYSA-N 3-thiophen-2-ylpyrrole-2,5-dione Chemical compound O=C1NC(=O)C(C=2SC=CC=2)=C1 AUDYZXNUHIIGRB-UHFFFAOYSA-N 0.000 description 5
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 description 5
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000000692 anti-sense effect Effects 0.000 description 5
- 125000004432 carbon atom Chemical group C* 0.000 description 5
- 150000004676 glycans Chemical class 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 238000002560 therapeutic procedure Methods 0.000 description 5
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 4
- 241001529936 Murinae Species 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
- 230000022534 cell killing Effects 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 238000003818 flash chromatography Methods 0.000 description 4
- 230000002949 hemolytic effect Effects 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- 210000003712 lysosome Anatomy 0.000 description 4
- 230000001868 lysosomic effect Effects 0.000 description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 239000002539 nanocarrier Substances 0.000 description 4
- 239000002105 nanoparticle Substances 0.000 description 4
- 238000007747 plating Methods 0.000 description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- 229940126586 small molecule drug Drugs 0.000 description 4
- 150000003573 thiols Chemical class 0.000 description 4
- YWWDBCBWQNCYNR-UHFFFAOYSA-N trimethylphosphine Chemical compound CP(C)C YWWDBCBWQNCYNR-UHFFFAOYSA-N 0.000 description 4
- 230000035899 viability Effects 0.000 description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 3
- QRZUPJILJVGUFF-UHFFFAOYSA-N 2,8-dibenzylcyclooctan-1-one Chemical compound C1CCCCC(CC=2C=CC=CC=2)C(=O)C1CC1=CC=CC=C1 QRZUPJILJVGUFF-UHFFFAOYSA-N 0.000 description 3
- 108020000948 Antisense Oligonucleotides Proteins 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 238000010461 azide-alkyne cycloaddition reaction Methods 0.000 description 3
- 230000004888 barrier function Effects 0.000 description 3
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 3
- 239000012267 brine Substances 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- 125000003636 chemical group Chemical group 0.000 description 3
- 238000007385 chemical modification Methods 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 230000001086 cytosolic effect Effects 0.000 description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 3
- 125000000524 functional group Chemical group 0.000 description 3
- 230000037440 gene silencing effect Effects 0.000 description 3
- 210000005260 human cell Anatomy 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 239000002502 liposome Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- CHKVPAROMQMJNQ-UHFFFAOYSA-M potassium bisulfate Chemical compound [K+].OS([O-])(=O)=O CHKVPAROMQMJNQ-UHFFFAOYSA-M 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000013615 primer Substances 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 230000000630 rising effect Effects 0.000 description 3
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- OIGKWPIMJCPGGD-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-[2-[2-[2-(2-azidoethoxy)ethoxy]ethoxy]ethoxy]propanoate Chemical compound [N-]=[N+]=NCCOCCOCCOCCOCCC(=O)ON1C(=O)CCC1=O OIGKWPIMJCPGGD-UHFFFAOYSA-N 0.000 description 2
- RMZNXRYIFGTWPF-UHFFFAOYSA-N 2-nitrosoacetic acid Chemical group OC(=O)CN=O RMZNXRYIFGTWPF-UHFFFAOYSA-N 0.000 description 2
- SRLVISRRXRYXHV-UHFFFAOYSA-N 3-[2-[3-[2-[2-[2-(2-azidoethoxy)ethoxy]ethoxy]ethoxy]propanoylamino]-3-(2-carboxyethoxy)-2-(2-carboxyethoxymethyl)propoxy]propanoic acid Chemical compound OC(=O)CCOCC(COCCC(O)=O)(COCCC(O)=O)NC(=O)CCOCCOCCOCCOCCN=[N+]=[N-] SRLVISRRXRYXHV-UHFFFAOYSA-N 0.000 description 2
- HSJKGGMUJITCBW-UHFFFAOYSA-N 3-hydroxybutanal Chemical compound CC(O)CC=O HSJKGGMUJITCBW-UHFFFAOYSA-N 0.000 description 2
- NHWDRMRILVPSFR-VSMKPTLZSA-N 4-[(E)-[6-(2,5-dioxopyrrol-1-yl)hexanoylhydrazinylidene]methyl]-N-[2-[2-[2-[2-[3-[[4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl]methylamino]-3-oxopropoxy]ethoxy]ethoxy]ethoxy]ethyl]benzamide Chemical compound O=C1N(C(C=C1)=O)CCCCCC(=O)N\N=C\C1=CC=C(C(=O)NCCOCCOCCOCCOCCC(=O)NCC2=CC=C(C=C2)C=2N=NC(=NN=2)C)C=C1 NHWDRMRILVPSFR-VSMKPTLZSA-N 0.000 description 2
- OVRUOPINRDOPSA-UJGPHIHISA-N C(C)(=O)O[C@H]1[C@H](O[C@H]([C@@H]([C@H]1OC(C)=O)NC(C)=O)OCCCCC(=O)ON1C(CCC1=O)=O)COC(C)=O Chemical compound C(C)(=O)O[C@H]1[C@H](O[C@H]([C@@H]([C@H]1OC(C)=O)NC(C)=O)OCCCCC(=O)ON1C(CCC1=O)=O)COC(C)=O OVRUOPINRDOPSA-UJGPHIHISA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 2
- 239000007821 HATU Substances 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 2
- 238000003820 Medium-pressure liquid chromatography Methods 0.000 description 2
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 description 2
- 229920002873 Polyethylenimine Polymers 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 2
- 238000002123 RNA extraction Methods 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- 238000003800 Staudinger reaction Methods 0.000 description 2
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 150000001408 amides Chemical class 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 125000004106 butoxy group Chemical group [*]OC([H])([H])C([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 2
- 125000002091 cationic group Chemical group 0.000 description 2
- 229920006317 cationic polymer Polymers 0.000 description 2
- 239000012230 colorless oil Substances 0.000 description 2
- XCEBOJWFQSQZKR-UHFFFAOYSA-N dbco-nhs Chemical compound C1C2=CC=CC=C2C#CC2=CC=CC=C2N1C(=O)CCC(=O)ON1C(=O)CCC1=O XCEBOJWFQSQZKR-UHFFFAOYSA-N 0.000 description 2
- 239000005547 deoxyribonucleotide Substances 0.000 description 2
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 2
- KZNICNPSHKQLFF-UHFFFAOYSA-N dihydromaleimide Natural products O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 description 2
- 238000007876 drug discovery Methods 0.000 description 2
- 230000002900 effect on cell Effects 0.000 description 2
- 239000012039 electrophile Substances 0.000 description 2
- 230000012202 endocytosis Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- 239000006260 foam Substances 0.000 description 2
- 125000000623 heterocyclic group Chemical group 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 125000005597 hydrazone group Chemical group 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 238000010348 incorporation Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 125000004433 nitrogen atom Chemical group N* 0.000 description 2
- 230000000269 nucleophilic effect Effects 0.000 description 2
- 230000009437 off-target effect Effects 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 230000032361 posttranscriptional gene silencing Effects 0.000 description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 description 2
- 125000002572 propoxy group Chemical group [*]OC([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 2
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 108020001580 protein domains Proteins 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 125000000548 ribosyl group Chemical group C1([C@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- 238000007142 ring opening reaction Methods 0.000 description 2
- 239000002924 silencing RNA Substances 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 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 2
- 229960002317 succinimide Drugs 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- WXDPDERWOTZNLB-UHFFFAOYSA-N tert-butyl 3-[2-[3-[2-[2-[2-(2-azidoethoxy)ethoxy]ethoxy]ethoxy]propanoylamino]-3-[3-[(2-methylpropan-2-yl)oxy]-3-oxopropoxy]-2-[[3-[(2-methylpropan-2-yl)oxy]-3-oxopropoxy]methyl]propoxy]propanoate Chemical compound CC(C)(C)OC(=O)CCOCC(COCCC(=O)OC(C)(C)C)(COCCC(=O)OC(C)(C)C)NC(=O)CCOCCOCCOCCOCCN=[N+]=[N-] WXDPDERWOTZNLB-UHFFFAOYSA-N 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- GPXMKARIFPDQNT-UHFFFAOYSA-N (4-nitrophenyl) 3-acetylsulfanylpropanoate Chemical compound CC(=O)SCCC(=O)OC1=CC=C([N+]([O-])=O)C=C1 GPXMKARIFPDQNT-UHFFFAOYSA-N 0.000 description 1
- MBQYGQMGPFNSAP-UHFFFAOYSA-N 2-[2-[2-(2-azidoethoxy)ethoxy]ethoxy]ethanol Chemical compound OCCOCCOCCOCCN=[N+]=[N-] MBQYGQMGPFNSAP-UHFFFAOYSA-N 0.000 description 1
- QOSGKJURLPIMKU-UHFFFAOYSA-N 2-[2-[2-[2-[3-[2-[2-[2-[2-(propylamino)ethoxy]ethoxy]ethoxy]ethoxy]propylamino]ethoxy]ethoxy]ethoxy]ethylcarbamic acid Chemical compound CCCNCCOCCOCCOCCOCCCNCCOCCOCCOCCNC(O)=O QOSGKJURLPIMKU-UHFFFAOYSA-N 0.000 description 1
- PBVAJRFEEOIAGW-UHFFFAOYSA-N 3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochloride Chemical compound Cl.OC(=O)CCP(CCC(O)=O)CCC(O)=O PBVAJRFEEOIAGW-UHFFFAOYSA-N 0.000 description 1
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- CIMKBXFSXWOMDK-IQZDNPOKSA-N 5-[(2R,3R,4R,5R,6R)-3-acetamido-4,5-diacetyloxy-6-(acetyloxymethyl)oxan-2-yl]oxypentanoic acid Chemical compound C(C)(=O)N[C@H]1[C@@H](O[C@@H]([C@@H]([C@@H]1OC(C)=O)OC(C)=O)COC(C)=O)OCCCCC(=O)O CIMKBXFSXWOMDK-IQZDNPOKSA-N 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000244203 Caenorhabditis elegans Species 0.000 description 1
- 101100454807 Caenorhabditis elegans lgg-1 gene Proteins 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- AVGPOAXYRRIZMM-UHFFFAOYSA-N D-Apiose Natural products OCC(O)(CO)C(O)C=O AVGPOAXYRRIZMM-UHFFFAOYSA-N 0.000 description 1
- ASNHGEVAWNWCRQ-LJJLCWGRSA-N D-apiofuranose Chemical compound OC[C@@]1(O)COC(O)[C@@H]1O ASNHGEVAWNWCRQ-LJJLCWGRSA-N 0.000 description 1
- ASNHGEVAWNWCRQ-UHFFFAOYSA-N D-apiofuranose Natural products OCC1(O)COC(O)C1O ASNHGEVAWNWCRQ-UHFFFAOYSA-N 0.000 description 1
- 108020001019 DNA Primers Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 101000610640 Homo sapiens U4/U6 small nuclear ribonucleoprotein Prp3 Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 101710203526 Integrase Proteins 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 238000000719 MTS assay Methods 0.000 description 1
- 231100000070 MTS assay Toxicity 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- CIGXGWDUXOBPIO-UHFFFAOYSA-N OC(C=C1)NC1=O Chemical compound OC(C=C1)NC1=O CIGXGWDUXOBPIO-UHFFFAOYSA-N 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- 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 1
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 1
- 241000243142 Porifera Species 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 101001110823 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) 60S ribosomal protein L6-A Proteins 0.000 description 1
- 101000712176 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) 60S ribosomal protein L6-B Proteins 0.000 description 1
- 239000012163 TRI reagent Substances 0.000 description 1
- 108010033576 Transferrin Receptors Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 102100040374 U4/U6 small nuclear ribonucleoprotein Prp3 Human genes 0.000 description 1
- KPFBUSLHFFWMAI-HYRPPVSQSA-N [(8r,9s,10r,13s,14s,17r)-17-acetyl-6-formyl-3-methoxy-10,13-dimethyl-1,2,7,8,9,11,12,14,15,16-decahydrocyclopenta[a]phenanthren-17-yl] acetate Chemical compound C1C[C@@H]2[C@](CCC(OC)=C3)(C)C3=C(C=O)C[C@H]2[C@@H]2CC[C@](OC(C)=O)(C(C)=O)[C@]21C KPFBUSLHFFWMAI-HYRPPVSQSA-N 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 150000001541 aziridines Chemical class 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 238000010805 cDNA synthesis kit Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000011284 combination treatment Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- NHFQNAGPXIVKND-UHFFFAOYSA-N dbco-maleimide Chemical compound C1C2=CC=CC=C2C#CC2=CC=CC=C2N1C(=O)CCNC(=O)CCN1C(=O)C=CC1=O NHFQNAGPXIVKND-UHFFFAOYSA-N 0.000 description 1
- 229940124447 delivery agent Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000005906 dihydroxylation reaction Methods 0.000 description 1
- UXGNZZKBCMGWAZ-UHFFFAOYSA-N dimethylformamide dmf Chemical compound CN(C)C=O.CN(C)C=O UXGNZZKBCMGWAZ-UHFFFAOYSA-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
- 231100000676 disease causative agent Toxicity 0.000 description 1
- 125000002228 disulfide group Chemical group 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 230000002121 endocytic effect Effects 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006735 epoxidation reaction Methods 0.000 description 1
- 150000002118 epoxides Chemical class 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 1
- VFRSADQPWYCXDG-LEUCUCNGSA-N ethyl (2s,5s)-5-methylpyrrolidine-2-carboxylate;2,2,2-trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.CCOC(=O)[C@@H]1CC[C@H](C)N1 VFRSADQPWYCXDG-LEUCUCNGSA-N 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000010363 gene targeting Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 150000004820 halides Chemical class 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000010468 interferon response Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- FSQQTNAZHBEJLS-UPHRSURJSA-N maleamic acid Chemical compound NC(=O)\C=C/C(O)=O FSQQTNAZHBEJLS-UPHRSURJSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108020004084 membrane receptors Proteins 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- MBLBDJOUHNCFQT-OSMVPFSASA-N n-[(2r,3r,4r,5r)-3,4,5,6-tetrahydroxy-1-oxohexan-2-yl]acetamide Chemical compound CC(=O)N[C@@H](C=O)[C@@H](O)[C@@H](O)[C@H](O)CO MBLBDJOUHNCFQT-OSMVPFSASA-N 0.000 description 1
- WOOWBQQQJXZGIE-UHFFFAOYSA-N n-ethyl-n-propan-2-ylpropan-2-amine Chemical compound CCN(C(C)C)C(C)C.CCN(C(C)C)C(C)C WOOWBQQQJXZGIE-UHFFFAOYSA-N 0.000 description 1
- SQDFHQJTAWCFIB-UHFFFAOYSA-N n-methylidenehydroxylamine Chemical compound ON=C SQDFHQJTAWCFIB-UHFFFAOYSA-N 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 108091027963 non-coding RNA Proteins 0.000 description 1
- 102000042567 non-coding RNA Human genes 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 239000000123 paper Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229960003531 phenolsulfonphthalein Drugs 0.000 description 1
- 150000008300 phosphoramidites Chemical class 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000004627 regenerated cellulose Substances 0.000 description 1
- 230000000754 repressing effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- NWMIYTWHUDFRPL-UHFFFAOYSA-N sapogenin Natural products COC(=O)C1(CO)C(O)CCC2(C)C1CCC3(C)C2CC=C4C5C(C)(O)C(C)CCC5(CCC34C)C(=O)O NWMIYTWHUDFRPL-UHFFFAOYSA-N 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 125000005629 sialic acid group Chemical group 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000001839 systemic circulation Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- KPXODPHVWBQJEG-UHFFFAOYSA-N tert-butyl 3-[2-amino-3-[3-[(2-methylpropan-2-yl)oxy]-3-oxopropoxy]-2-[[3-[(2-methylpropan-2-yl)oxy]-3-oxopropoxy]methyl]propoxy]propanoate Chemical compound CC(C)(C)OC(=O)CCOCC(N)(COCCC(=O)OC(C)(C)C)COCCC(=O)OC(C)(C)C KPXODPHVWBQJEG-UHFFFAOYSA-N 0.000 description 1
- POHWAQLZBIMPRN-UHFFFAOYSA-N tert-butyl n-(3-aminopropyl)carbamate Chemical compound CC(C)(C)OC(=O)NCCCN POHWAQLZBIMPRN-UHFFFAOYSA-N 0.000 description 1
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000036642 wellbeing Effects 0.000 description 1
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
- A61K47/51—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
- A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- 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
- A61K47/554—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 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
Definitions
- the invention relates to a pharmaceutical combination comprising: a conjugate of an effector molecule and a ligand for ASGPR, wherein the ligand for ASGPR comprises at least one GalNAc moiety; and a saponin of the monodesmosidic or bidesmosidic triterpene glycoside type.
- the invention also relates to a pharmaceutical composition comprising the conjugate and the saponin.
- the invention relates to a pharmaceutical combination or composition of the invention, for use as a medicament, or for use in the treatment or prophylaxis of a disease or health problem in which an expression product is involved of any one or more of genes: apoB, TTR, PCSK9, ALAS1 , AT3, GO, CC5, X gene of HBV, S gene of HBV, AAT and LDH, and/or for use in the treatment or prophylaxis of a cancer, an infectious disease, a viral infection, hypercholesterolemia, primary hyperoxaluria, haemophilia A, haemophilia B, AAT related liver disease, acute hepatic porphyria, TTR-mediated amyloidosis, hereditary TTR amyloidosis (hATTR), complement-mediated disease, hepatitis B infection, or an auto-immune disease.
- genes apoB, TTR, PCSK9, ALAS1 , AT3, GO, CC5, X gene of H
- the invention relates to an in vitro or ex vivo method for transferring an effector molecule of the invention from outside a cell to inside said cell, preferably into the cytosol of said cell.
- the invention also relates to an in vitro or ex vivo method for transferring the conjugate of the invention from outside a cell to inside said cell.
- Oligonucleotide therapy is a relatively young and fast-developing field which aims to overcome many of the issues encountered with small-molecule drugs or antibody drugs by directly manipulating the genetic transcription and translation pathways.
- the potency and versatility of oligonucleotides in particular the prospect of suppressing genes encoding proteins that are ‘undruggable’ by classical small molecule drugs, makes them attractive drug candidates.
- the first oligonucleotide drugs were based on antisense technology, whereby single-stranded nucleic acid molecules would bind with sequence specificity to their complementary mRNA target, thus triggering degradation of the duplex by the RNase H system.
- RNAi double-stranded RNAs
- dsRNAs double-stranded RNAs
- PTGS post-transcriptional gene silencing
- RNAi RNA interference
- RNAi pathway through small dsRNAs such as small interfering RNAs (siRNAs) and short hairpin RNA (shRNA) has several theoretical advantages over antisense being notably more efficient (catalytic) and giving longer inhibition of gene expression. This could translate into lower doses and lower cost together with less frequent dosing. Lower exposures could also mean fewer toxicity problems for siRNA.
- siRNAs small interfering RNAs
- shRNA short hairpin RNA
- siRNA RNA-lnduced Silencing Complex
- RISC RNA-lnduced Silencing Complex
- the siRNA Upon entering the bloodstream, siRNA is vulnerable to degradation by endogenous nucleases, and renal excretion due to its small size and highly anionic character.
- the siRNA before reaching its target cell, the siRNA must navigate the tight endothelial junctions of the blood vessels and diffuse through the extracellular matrix. Due to its numerous negative charges, siRNA does not readily bind to or cross the cell membrane, and once inside cells, it must escape from endosomes to interact with its intracellular protein targets.
- oligonucleotide such as siRNA
- delivery systems which improve oligonucleotide (such as siRNA) delivery.
- the main methods employed to enhance siRNA (or other oligonucleotides) delivery to the cell employ liposomes, cell-penetrating peptides (CPPs) and their mimics, or nanoparticles (Gooding, Matt, et al. Chemical biology & drug design 80.6 (2012): 787-809).
- CPPs cell-penetrating peptides
- Safety concerns such as the possibility of insertion mutagenesis and immunogenesis, are considered to limit the future of viral approaches.
- Liposomes are the most commonly used delivery vector for oligonucleotides such as siRNA, where the oligonucleotide is encapsulated within a lipid bilayer.
- oligonucleotides such as siRNA
- Several problems are encountered with liposomal oligonucleotide delivery, such as oxygen radical-mediated toxicity (typical for of cationic liposomes), cell toxicity, effects on gene regulation and inflammatory responses.
- oxygen radical-mediated toxicity typically of cationic liposomes
- cell toxicity effects on gene regulation and inflammatory responses.
- in vivo delivery using lipids seems to predominantly target the liver and spleen.
- CPP Cell-penetrating peptides
- protein transduction domains are short peptides (usually ⁇ 30 amino-acid residues long) which have the unusual property of being able to cross the cell membrane.
- a CPP may enhance cellular uptake of the oligonucleotide.
- the field of CPP mediated oligonucleotide delivery is extremely complex since it combines the challenges posed by both oligonucleotide technology and peptide technology, two fields which are far from mature, in a single drug.
- Nanoparticles and nanocarriers are nanoscale oligonucleotide delivery systems typically comprised of a polymer, biological stabilizers and cell-targeting ligands complexed with the oligonucleotide.
- An exemplary siRNA nanocarrier system is known under the name siRNA Dynamic Poly-Conjugates. This system is based around an amphipathic polymer linked to polyethylene glycol (PEG) as a biological stabilizer and a hepatocyte-targeting ligand wherein the siRNA is covalently bound to the polymer by a disulfide bond.
- PEG polyethylene glycol
- the targeting moieties and PEG moieties are attached to the polymer via maleamate linkages, forming negatively charged nanoparticles, which do not bind to serum proteins.
- Nanocarriers are also known in the form of cationic polymers such as polyethyleneimines (PEIs) (sometimes combined with cyclodextrins), which can form electrostatic complexes with oligonucleotides such as siRNA, affording protection from degradation and aiding internalisation.
- PEIs polyethyleneimines
- oligonucleotides such as siRNA
- oligonucleotide such as siRNA
- endosomal escape remains a major barrier to the application of oligonucleotide-based therapeutics such as RNAi-based therapeutics.
- oligonucleotide-based therapeutics such as RNAi-based therapeutics.
- Recent literature suggests a passive siRNA escape rate of ⁇ 0.01% (Setten, Ryan L, et al. Nature Reviews Drug Discovery 18.6 (2019): 421 -446).
- An emerging strategy entails the conjugation of antisense oligonucleotides (ASOs) to receptor ligands in order to increase oligonucleotide potency and distribution to selected tissues.
- ASOs antisense oligonucleotides
- GalNAc triantennary N-acetyl galactosamine
- GalNAc is found on damaged glycoproteins that have lost terminal sialic acid residues from their pendant oligosaccharides.
- the liver functions to clear these proteins from the systemic circulation by expressing trimeric asialoglycoprotein receptors (ASGPRs), preferably ASGPR1 , at very high levels (order of 10 s — 10 6 per cell) on the surface of hepatocytes.
- ASGPRs bind specifically to GalNAc at neutral pH for endocytosis of circulating macromolecules from the blood and release GalNAc at acidic pH ( ⁇ 5-6) for cargo drop off in the early endosome. Freed ASGPRs are then recycled back to the cell surface for reuse.
- RNAi drugs currently in clinical trials are single- molecule, chemically modified RNAi triggers conjugated to multivalent GalNAc ligands targeting the ASGPRs.
- the suitable physiology of the liver, the unique properties of ASGPRs, the non- toxic nature of the GalNAc ligand and the simplicity of GalNAc-siRNA conjugates make this an attractive approach for systemic RNAi delivery to hepatocytes.
- a first aspect of the invention relates to a pharmaceutical combination comprising:
- ligand for ASGPR comprises at least one /V-acetylgalactosamine (GalNAc) moiety, preferably three or four GalNAc moieties, more preferably the ligand for ASGPR comprises or consists of (GalNAc)3Tris; and
- saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside, and optionally comprising a pharmaceutically acceptable excipient and/or pharmaceutically acceptable diluent.
- composition comprising the saponin and optionally comprising a pharmaceutically acceptable excipient and/or pharmaceutically acceptable diluent.
- An embodiment is the pharmaceutical combination of the invention in the form of a single pharmaceutical composition comprising the conjugate, the saponin and optionally a pharmaceutically acceptable excipient and/or pharmaceutically acceptable diluent.
- a second aspect of the invention relates to the pharmaceutical combination of the invention, for use as a medicament.
- a third aspect of the invention relates to the pharmaceutical combination of the invention, for use in the treatment or prophylaxis of a disease or health problem in which an expression product is involved of any one or more of genes: apoB, TTR, PCSK9, ALAS1 , AT3, GO, CC5, X gene of HBV, S gene of HBV, AAT and LDH.
- An embodiment is the pharmaceutical combination for use according to the invention, wherein said use is in the treatment or prophylaxis of a cancer, an infectious disease, a viral infection, hypercholesterolemia, primary hyperoxaluria, haemophilia A, haemophilia B, AAT related liver disease, acute hepatic porphyria, TTR-mediated amyloidosis, hereditary TTR amyloidosis (hATTR), complement-mediated disease, hepatitis B infection, or an auto-immune disease.
- a fourth aspect of the invention relates to an in vitro or ex vivo method for transferring an effector molecule of the invention from outside a cell to inside said cell, preferably into the cytosol of said cell, comprising the steps of: a) providing a cell which expresses ASGPR, preferably ASGPR1 , on its surface, preferably selected from a liver cell, a virally infected cell and a cancer cell; b) providing the conjugate of the invention, the conjugate comprising the effector molecule to be transferred; c) providing the saponin of the invention; d) contacting the cell of step a) in vitro or ex vivo with the conjugate of step b) and the saponin of step c), therewith effecting the transfer of said conjugate comprising the effector molecule from outside the cell to inside said cell, and by effecting the transfer of said conjugate effecting the transfer of the effector molecule from outside the cell to inside said cell, preferably into the cytosol of said cell.
- a fifth aspect of the invention relates to an in vitro or ex vivo method for transferring the conjugate of the invention from outside a cell to inside said cell, preferably into the cytosol of said cell, comprising the steps of: a) providing a cell which expresses ASGPR, preferably ASGPR1 , on its surface, preferably selected from a liver cell, a virally infected cell and a cancer cell; b) providing the conjugate of the invention; c) providing the saponin of the invention; d) contacting the cell of step a) in vitro or ex vivo with the conjugate of step b) and the saponin of step c), therewith effecting the transfer of the conjugate from outside the cell to inside said cell.
- a sixth aspect of the invention relates to a kit of parts, comprising the pharmaceutical combination of the invention or the second pharmaceutical composition of the invention, and instructions for use of said pharmaceutical combination or second pharmaceutical composition according to the invention or for use in an in vitro or ex vivo method according to the invention.
- GalNAc has its regular scientific meaning and here refers to N-acetylgalactosamine and to the lUPAC name thereof: 2-(acetylamino)-2-deoxy-D-galactose.
- (GalNAc)3Tris has its regular scientific meaning in for example the field of siRNA- based therapy, and here refers to a moiety comprising three GalNAc units each separately covalently bound to the hydroxyl groups of tris(hydroxymethyl)aminomethane (Tris) (lUPAC name: 2-amino-2- (hydroxymethyl) propane-1 ,3-diol), preferably via at least one linker.
- Tris tris(hydroxymethyl)aminomethane
- (GalNAc)3Tris can exist as a free amine comprising molecule or may be further functionalized via the remaining amine binding site, for example to form the (GalNAc)3Tris-moiety comprising conjugates described herein.
- oligonucleotide has its regular scientific meaning and here refers to a string of two or more nucleotides, i.e. an oligonucleotides is a short oligomer composed of ribonucleotides or deoxyribonucleotides.
- nucleic acids such as RNA and DNA, and any modified RNA or DNA, such as a string of nucleic acids comprising a nucleotide analogue such as a bridged nucleic acid (BNA), also known as locked nucleic acid (LNA) or a 2'-0,4'-C-aminoethylene or a 2'-0,4'-C- aminomethylene bridged nucleic acid (BNA NC ), wherein the nucleotide is a ribonucleotide or a deoxyribonucleotide.
- BNA bridged nucleic acid
- LNA locked nucleic acid
- BNA NC 2'-0,4'-C-aminoethylene
- the nucleotide is a ribonucleotide or a deoxyribonucleotide.
- RNAi-mediated gene-targeting has its regular scientific meaning and here refers to the in vivo, ex vivo or in vitro approach of influencing functioning of the gene in a cell by transferring into said cell an oligonucleotide, such as a small double-stranded RNA molecule, that targets mRNA involved in transcription of the gene: for example, small double-stranded RNA molecules can efficiently trigger RNAi silencing of specific genes.
- bridged nucleic acid in short, or “locked nucleic acid” or “LNA” in short or 2'-0,4'-C-aminoethylene or 2'-0,4'-C-aminomethylene bridged nucleic acid (BNA NC ), has its regular scientific meaning and here refers to a modified RNA nucleotide.
- a BNA is also referred to as ‘constrained RNA molecule’ or ‘inaccessible RNA molecule’.
- a BNA monomer can contain a five- membered, six-membered or even a seven-membered bridged structure with a “fixed” C3’-endo sugar puckering.
- the bridge is synthetically incorporated at the 2’, 4’-position of the ribose to afford a 2’, 4’- BNA monomer.
- a BNA monomer can be incorporated into an oligonucleotide polymeric structure using standard phosphoramidite chemistry known in the art.
- a BNA is a structurally rigid oligonucleotide with increased binding affinity and stability.
- antisense oligonucleotide has its regular scientific meaning and may be indicated in short in the description as “AON” or “ASO”.
- BNA-based antisense oligonucleotide or in short “BNA-AON”, has its regular scientific meaning and here refers to a string of antisense nucleotides wherein at least one of said nucleotides is a BNA.
- proteinaceous has its regular scientific meaning and here refers to a molecule that is protein-like, meaning that the molecule possesses, to some degree, the physicochemical properties characteristic of a protein, is of protein, relating to protein, containing protein, pertaining to protein, consisting of protein, resembling protein, or being a protein.
- proteinaceous as used in for example ‘proteinaceous molecule’ refers to the presence of at least a part of the molecule that resembles or is a protein, wherein ‘protein’ is to be understood to include a chain of amino-acid residues at least two residues long, thus including a peptide, a polypeptide and a protein and an assembly of proteins or protein domains.
- the at least two amino-acid residues are for example linked via (an) amide bond(s), such as (a) peptide bond(s).
- the amino- acid residues are natural amino-acid residues and/or artificial amino-acid residues such as modified natural amino-acid residues.
- a proteinaceous molecule is a molecule comprising at least two amino-acid residues, preferably between two and about 2.000 amino-acid residues.
- a proteinaceous molecule is a molecule comprising from 2 to 20 (typical for a peptide) amino acids.
- a proteinaceous molecule is a molecule comprising from 21 to 1 .000 amino acids (typical for a polypeptide, a protein, a protein domain, such as an antibody, a single domain antibody, a Fab, an scFv, a ligand for a receptor such as EGF).
- the amino- acid residues are (typically) linked via (a) peptide bond(s).
- said amino-acid residues are or comprise (modified) (non-)natural amino acid residues.
- effector molecule when referring to the effector molecule as part of e.g. a covalent conjugate, has its regular scientific meaning and here refers to a molecule that can selectively bind to for example any one or more of the target molecules: a protein, a peptide, a carbohydrate, a saccharide such as a glycan, a (phospho)lipid, a nucleic acid such as DNA, RNA, an enzyme, and regulates the biological activity of such one or more target molecule(s).
- the effector molecule is for example a molecule selected from any one or more of a small molecule such as a drug molecule, a toxin such as a protein toxin, an oligonucleotide such as a BNA, a xeno nucleic acid or an siF!NA, an enzyme, a peptide, a protein, or any combination thereof.
- a small molecule such as a drug molecule
- a toxin such as a protein toxin
- an oligonucleotide such as a BNA, a xeno nucleic acid or an siF!NA
- an enzyme a peptide, a protein, or any combination thereof.
- an effector molecule or an effector moiety is a molecule or moiety selected from anyone or more of a small molecule such as a drug molecule, a toxin such as a protein toxin, an oligonucleotide such as a BNA, a xeno nucleic acid or an siF!NA, an enzyme, a peptide, a protein, or any combination thereof, that can selectively bind to any one or more of the target molecules: a protein, a peptide, a carbohydrate, a saccharide such as a glycan, a (phospho)lipid, a nucleic acid such as DNA, RNA, an enzyme, and that upon binding to the target molecule regulates the biological activity of such one or more target molecule(s).
- a small molecule such as a drug molecule
- a toxin such as a protein toxin
- an oligonucleotide such as a BNA, a xeno nucleic acid
- an effector molecule can exert a biological effect inside a cell such as a mammalian cell such as a human cell, such as in the cytosol of said cell.
- Typical effector molecules are thus drug molecules, plasmid DNA, toxins such as toxins comprised by antibody-drug conjugates (ADCs), oligonucleotides such as siRNA, BNA, nucleic acids comprised by an antibody-oligonucleotide conjugate (AOC).
- ADCs antibody-drug conjugates
- oligonucleotides such as siRNA, BNA
- AOC antibody-oligonucleotide conjugate
- an effector molecule is a molecule which can act as a ligand that can increase or decrease (intracellular) enzyme activity, gene expression, or cell signalling.
- health problem has its regular scientific meaning and here refers to any condition of the body of a subject such as a human patient, or of a part, organ, muscle, vein, artery, skin, limb, blood, cell, etc. thereof, that is suboptimal when compared to the condition of that body or part thereof in a healthy subject, therewith hampering the proper functioning and/or well-being of the subject, e.g. impairs normal functioning of a human body.
- GalNAc-decorated oligonucleotide drug has its regular scientific meaning and here refers to an oligonucleotide for interfering in transcription of a gene, wherein one or more GalNAc units are coupled to the oligonucleotide, e.g. via at least one linker.
- locked nucleic acid in short “LNA” has its regular scientific meaning and here refers to an oligonucleotide that contains one or more nucleotide building blocks in which an additional methylene bridge fixes the ribose moiety either in the C3'-endo conformation (beta-D-LNA) or C2'-endo (alpha-L-LNA) conformation, as is known in the art.
- bridged nucleic acid NC in short “BNA NC ” has its regular scientific meaning and here refers to an oligonucleotide that contains one or more nucleotide building blocks in which an additional 2'-0,4'-C-aminoethylene bridged nucleic acid is comprised, with different substitutions at the N atom (of which a methyl group is the most commonly used to date), as is known in the art.
- siRNA has its regular scientific meaning and here refers to an siRNA molecule comprising chemical modifications compared to the RNA oligonucleotide consisting of naturally occurring nucleotides, such that the modified siRNA molecule is more resistant towards metabolic degradation, digestion, enzymatic lysis, etc., i.e. more stable.
- the term “saponin” has its regular scientific meaning and here refers to a group of amphipatic glycosides which comprise one or more hydrophilic glycone moieties combined with a lipophilic aglycone core which is a sapogenin.
- the saponin may be naturally occurring or synthetic (i.e. non-naturally occurring).
- the term “saponin” includes naturally occurring saponins, derivatives of naturally occurring saponins as well as saponins synthesized de novo through chemical and/or biotechnological synthesis routes.
- saponin derivative also known as “modified saponin”
- saponin derivative has its regular scientific meaning and here refers to a compound corresponding to a naturally-occurring saponin which has been derivatised by one or more chemical modifications, such as the oxidation of a functional group, the reduction of a functional group and/or the formation of a covalent bond with another molecule (also referred to as “conjugation” or as “covalent conjugation”).
- Preferred modifications include derivatisation of an aldehyde group of the aglycone core; of a carboxyl group of a saccharide chain or of an acetoxy group of a saccharide chain.
- the saponin derivative does not have a natural counterpart, i.e.
- saponin derivative is not produced naturally by e.g. plants or trees.
- saponin derivative includes derivatives obtained by derivatisation of naturally-occurring saponins as well as derivatives synthesized de novo through chemical and/or biotechnological synthesis routes resulting in a compound corresponding to a naturally occurring saponin which has been derivatised by one or more chemical modifications.
- aglycone core structure has its regular scientific meaning and here refers to the aglycone core, or aglycone, of a saponin without the one or two carbohydrate antenna or saccharide chains (glycans, glycones) bound thereto.
- quillaic acid is the aglycone core structure for S01861 , QS-7 and QS-21.
- saccharide chain has its regular scientific meaning and here refers to any of a glycan, a carbohydrate antenna, a glycone, a single saccharide moiety (monosaccharide) or a chain comprising multiple saccharide moieties (oligosaccharide, polysaccharide).
- the saccharide chain can consist of only saccharide moieties or may also comprise further moieties such as any one of 4E-Methoxycinnamic acid, 4Z-Methoxycinnamic acid, and 5-0-[5-0-Ara/Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy- 6-methyl-octanoic acid), such as for example present in QS-21 .
- Api/Xyl-“ or “Api- or Xyl-“ in the context of the name of a saccharide chain has its regular scientific meaning and here refers to the saccharide chain either comprising an apiose (Api) moiety, or comprising a xylose (Xyl) moiety.
- antibody-drug conjugate has its regular scientific meaning and here refers to any conjugate of an antibody such as an IgG, a Fab, an scFv, a single domain antibody, an immunoglobulin, an immunoglobulin fragment, one or multiple Vh domains, etc., with any molecule that can exert a therapeutic effect when contacted with cells of a subject such as a human patient, such as an active pharmaceutical ingredient, a toxin, an oligonucleotide, an enzyme, a small molecule drug compound, etc.
- ADC antibody-drug conjugate
- antibody-oligonucleotide conjugate has its regular scientific meaning and here refers to any conjugate of an antibody such as an IgG, a Fab, a single domain antibody, an scFv, an immunoglobulin, an immunoglobulin fragment, one or multiple Vh domains, etc., and any oligonucleotide molecule that can exert a therapeutic effect when contacted with cells of a subject such as a human patient, such as an oligonucleotide selected from a natural or synthetic string of nucleic acids encompassing DNA, modified DNA, RNA, modified RNA, synthetic nucleic acids, presented as a single-stranded molecule or a double-stranded molecule, such as a BNA, an allele-specific oligonucleotide, a short or small interfering RNA (siRNA; silencing RNA), an anti-sense DNA, anti-sense RNA, etc.
- siRNA silencing RNA
- conjugate has its regular scientific meaning and here refers to at least a first molecule that is covalently bound through (a) chemical bond(s) to at least a second molecule, therewith forming an covalently coupled assembly comprising or consisting of the first molecule and the second molecule.
- Typical conjugates are (GalNAc)3Tris - siRNA, an ADC, an AOC, and S01861 -EMCFI (EMCFI linked to the aldehyde group of the aglycone core structure of the saponin).
- S as used such as in a GalNAc - effector molecule conjugate or construct comprising a linker, represents ‘stable linker’ which remains intact in the endosome and in the cytosol.
- L as used such as in an antibody-saponin conjugate or construct comprising a linker, represents ‘labile linker’ which is cleaved under slightly acid conditions in the endosome.
- moiety has its regular scientific meaning and here refers to a molecule that is bound, linked, conjugated to a further molecule, linker, assembly of molecules, etc., and therewith forming part of a larger molecule, conjugate, assembly of molecules.
- a moiety is a first molecule that is covalently bound to a second molecule, involving one or more chemical groups initially present on the first molecule and present on the second molecule.
- saporin is a typical molecule, and is an example of an effector molecule.
- the saporin is a typical effector moiety in the ADC.
- a BNA or an siRNA is a typical effector moiety in the AOC.
- DAR normally stands for Drug Antibody Ratio and refers to the average drug to antibody ratio for a given preparation of antibody drug conjugate (ADC) and here refers to a ratio of the amount of bound S01861 moieties, or SPT001 , or bound ApoBBNA with respect to the conjugate molecule.
- ADC antibody drug conjugate
- compositions comprising components A and B
- the only enumerated components of the composition are A and B, and further the claim should be interpreted as including equivalents of those components.
- indefinite article “a” or “an” does not exclude the possibility that more than one of the element or component are present, unless the context clearly requires that there is one and only one of the elements or components.
- the indefinite article “a” or “an” thus usually means “at least one”.
- Figure 1 synthesis of trivalent-GalNAc.
- Figure 3 synthesis of monovalent-GalNAc-S01861 .
- Figure 4 synthesis of trivalent-GalNAc-S01861 .
- Figure 5 synthesis of monovalent-GalNAc-BNA.
- Figure 8A and 8B synthesis of trivalent GalNAc.
- Figure 9A and 9B Cell viability (MTS) of GalNAc-S01861 and trivalent-GalNAc-S01861 on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 9B also applies for Figure 9A.
- Figure 10A and 10B Cell viability assay (MTS) HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 10B also applies for Figure 10A.
- Figure 11 A and 11B Gene expression analysis on HepG2 (A) and Huh7 (B) cell lines.
- Figure 12A and 12B Gene expression analysis on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 12B also applies for Figure 12A.
- Figure 13A and 13B Cell viability assay on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 13B also applies for Figure 13A.
- Figure 14A and B Gene expression analysis on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 14B also applies for Figure 14A.
- Figure 15A and B Cell viability assay on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 15B also applies for Figure 15A.
- Figure 16A and B Gene expression analysis on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 16B also applies for Figure 16A.
- Figure 17A and B Gene expression analysis on HepG2 (A) and Huh7 (B) cell lines. The legend displayed next to Figure 17B also applies for Figure 17A.
- Figure 18A and B ApoB RNA expression analysis and cell viability assay on human primary hepatocytes.
- Figure 19A and B ApoB RNA expression analysis and cell viability assay on human primary hepatocytes.
- Figure 20A and B ApoB RNA expression analysis assay on mouse primary hepatocytes.
- Figure 22 (GalNAc)3-Ls-BNA synthesis, intermediate 25, 26.
- ASGPR1 such as a GalNAc comprising ligand
- the therapeutic window of the conjugate is widened effectively.
- At least one of the above objectives is achieved by providing the therapeutic combination of at least two pharmaceutical compositions or the pharmaceutical composition of the invention.
- a first aspect of the invention relates to a pharmaceutical combination comprising:
- ligand for ASGPR comprises at least one /V-acetylgalactosamine (GalNAc) moiety, preferably three or four GalNAc moieties, more preferably the ligand for ASGPR comprises or consists of (GalNAc)3Tris; and
- saponin is a monodesmosidic triterpene glycoside or a bidesmosidic triterpene glycoside, and optionally comprising a pharmaceutically acceptable excipient and/or pharmaceutically acceptable diluent.
- composition comprising the saponin and optionally comprising a pharmaceutically acceptable excipient and/or pharmaceutically acceptable diluent.
- An embodiment is the pharmaceutical combination of the invention in the form of a single pharmaceutical composition comprising the conjugate, the saponin and optionally a pharmaceutically acceptable excipient and/or pharmaceutically acceptable diluent.
- a conjugate comprising a ligand for ASGPR and an effector molecule
- a conjugate comprising a ligand for ASGPR and an effector molecule
- the inventors now provide a solution for the long-felt need for improving the efficacy of delivery of e.g. oligonucleotides from outside a target cell to be treated with such an oligonucleotide, to inside said cell, in particular for improving the intracellularly delivery into the cytosol of a target cell that expresses ASGPR1 on its surface, of an oligonucleotide such as a BNA and/or an siRNA.
- the applied saponins according to the invention are known for their ability to enhance the endosomal escape of e.g.
- ADC antibody-drug conjugate
- such saponin derivatives are capable of enhancing the delivery of the effector molecule comprised by the conjugate into a target cell bearing the ASGPR1 , while the cytotoxicity and the haemolytic activity of such saponin derivates are lower than observed for the (naturally occurring) non-modified saponin.
- saponin derivatives are described in more detail herein.
- the pharmaceutical combination and certain pharmaceutical compositions of the present invention comprise a conjugate of an effector molecule and a ligand for asialoglycoprotein receptor (ASGPR) such as ASGPR1 , and preferably ASGPR1.
- ASGPR asialoglycoprotein receptor
- the ASGPR ligand comprises at least one N- acetylgalactosamine (GalNAc) moiety.
- each GalNAc moiety is bound to the remainder of the ASGPR ligand or - in case an ASGPR ligand consists of a single GalNAc moiety - to the effector moiety, via a covalent bond to the oxygen on position ⁇ ” as indicated in formula (I):
- the ASGPR ligand consists of a single GalNAc moiety bound to the effector moiety E, preferably via an effector moiety linker LE.
- the ASGPR ligand may comprise more than one GalNAc moiety, such as 2, 3, 4, 5 or 6 GalNAc moieties, preferably 3 or 4 GalNAc moieties, more preferably 3 GalNAc moieties.
- the GalNAc moieties are each separately covalently bound via the oxygen on position ⁇ ” to a central bridging moiety B, which effectively forms a bridge between the GalNAc moieties and the effector moiety, preferably via an effector moiety linker LE.
- the GalNAc moieties may be directly bound to the bridging moiety B as shown in formula (III): wherein n is an integer larger than or equal to 3 or 4, preferably n is 3, LE is an effector moiety linker and E is the effector moiety. More preferably, the GalNAc moieties are bound to the bridging moiety B via GalNAc linkers LGAL as shown in formula (IV):
- n is an integer larger than or equal to 3 or 4, preferably n is 3, LE is an effector moiety linker and E is the effector moiety. While each occurrence of LGAL may be independently chosen, the skilled person will appreciate that the synthesis of the conjugate is simplified in case each occurrence of LGAL represents the same moiety.
- the effector moiety linker LE represents any chemical moiety suitable for covalently binding an effector moiety to GalNAc as in formula (II) or to the bridging moiety B as in formula (III) and (IV).
- the identity and size of the linker is not particularly limited and covalently binding an effector molecule to GalNAc as in formula (II) or to the bridging moiety B as in formula (III) and (IV) may be effected by means of a ‘regular’ chemical functional group (e.g . an ether bond) as well as through “click-chemistry” type linkers which typically have a long chain length, resulting in a linker EL comprising e.g. more than 10 or more than 20 carbon atoms.
- Suitable effector moiety linkers LE and associated coupling reactions are described in the handbook Hermanson, Greg T. Bioconjugate techniques. Academic press, 2013.
- the effector moiety linker LE will typically be the result of a coupling reaction (e.g. “click-chemistry” type) between at least a first precursor LEI covalently bound to GalNAc or the bridging moiety B and a second precursor LE2 covalently bound to the effector moiety.
- a coupling reaction e.g. “click-chemistry” type
- LE is an effector moiety linker
- LEI is a precursor of the effector moiety linker LE which is covalently bound to GalNAc
- LE2 is a precursor of the effector moiety linker LE which is covalently bound to the effector moiety
- E is the effector moiety.
- LE is an effector moiety linker
- LEI is a precursor of the effector moiety linker LE which is covalently bound to GalNAc
- LE2 is a precursor of the effector moiety linker LE which is covalently bound to the effector moiety
- n is an integer larger than or equal to 3 or 4, preferably n is 3
- B is a bridging moiety
- E is the effector moiety.
- LE is an effector moiety linker
- LEI is a precursor of the effector moiety linker LE which is covalently bound to GalNAc
- LE2 is a precursor of the effector moiety linker LE which is covalently bound to the effector moiety
- n is an integer larger than or equal to 3 or 4, preferably n is 3
- E is the effector moiety
- LGAL is a GalNAc linker and B is a bridging moiety.
- the effector moiety linker LE can be the result of a coupling reaction between at least a first precursor LEI covalently bound to GalNAc or the bridging moiety and a second precursor LE2 covalently bound to the effector moiety, wherein the coupling reaction is for example an azide-alkyne cycloaddition, a thiol maleimide coupling, a staudinger reaction, a nucleophilic ring-opening of strained heterocyclic electrophiles (such as aziridines, epoxides, cyclic sulfates, aziridinium ions, episulfonium ions), a carbonyl reaction of the non-aldol type (such as urea, thiourea, hydrazon, oxime ether, amide or aromatic heterocycle formation) or an addition to a carbon-carbon double bond (such as epoxidation, aziridination, dihydroxylation, sulfentyl halide addition, nitros
- the effector moiety linker LE comprises a succinimide thioether moiety.
- a succinimide thioether is e.g. the result of a thiol maleimide coupling between an N-substituted maleimide and a thiol or sulfhydryl containing compound.
- This thiol maleimide coupling is a commonly known ‘click’-chemistry tool in the field of bioconjugation and is for example described in Hermanson, Greg T. Bioconjugate techniques. Academic press, 2013 page 289.
- the effector moiety linker LE is the result of a coupling reaction between a first precursor LEI covalently bound to GalNAc or the bridging moiety, the first precursor LEI comprising an N-substituted maleimide; and a second precursor LE2 covalently bound to the effector moiety, the second precursor LE2 comprising a thiol or a precursor thereof.
- a suitable thiol precursor is a disulfide, which may be cleaved ( e.g . in-situ) via reduction to the corresponding thiol.
- a preferred structure for the precursor LEI present in the compounds of formula (V), (VII) or (VIII) is the following terminal N-substituted maleimide:
- X represents any linker suitable for covalently bonding the terminal N-substituted maleimide to GalNAc or the bridging moiety B.
- X may be the result of a coupling reaction between a first moiety covalently bound to GalNAc or the bridging moiety and a second moiety covalently bound to the maleimide.
- X can comprise a hydrazone and/or a 1 , 2, 3-triazole. Whenever reference is made to a 1 , 2, 3-triazole in the context of the linkers of the present application, this preferably means a 1 H-1 , 2, 3-triazole.
- Embodiments of the structure for the precursor LEI present in the compounds of formula (V), (VII) or (VIII) are the following terminal N-substituted maleimides:
- a and b each independently represent an integer larger than or equal to 0, preferably a and b each independently represent an integer selected from 0, 1 , 2, and 3, more preferably a and b represent 2, and wherein LEI 3 represents a hydrazone and/or a 1 , 2, 3-triazole, preferably a 1 , 2, 3-triazole and wherein l_Eib represents a hydrazone and/or a 1 , 2, 3-triazole, preferably a hydrazone; wherein b and c each independently represent an integer larger than or equal to 0, preferably b represents an integer selected from 0, 1 , 2, and 3 and c represents an integer in the range of 5-15, more preferably b represents 2, and c represents 9, wherein l_Ei a represents a hydrazone and/or a 1 , 2, 3- triazole, preferably a 1 , 2, 3-triazole and wherein l_Eib represents a hydrazone and/or a 1 ,
- l_Eia, l-Eib and LEI C each preferably comprise less than 20 carbon atoms, more preferably l_Ei a , l_Eib and LEI C each independently represent a moiety according to formula (XIII), (XIV) or (XV), preferably l_Eia is the 1 ,2,3-triazole of formula (XIII), l_Eib is the hydrazone of formula (XIV) and LEI C is the 1 ,2,3- triazole of formula (XV) :
- the conjugate is a compound of formula (II), (III) or (IV) as described herein, wherein the effector moiety linker LE is the result of a coupling reaction between a compound of formula (V), (VII) or (VIII) with a compound of formula (VI), wherein the first precursor LEI is a terminal N-substituted maleimide of formula (X), (XI) or (XII), wherein LEI 3 is for example the 1 ,2,3-triazole of formula (XIII), l_Eib is for example the hydrazone of formula (XIV) and LEI C is for example the 1 ,2,3- triazole of formula (XV).
- the bridging moiety B present in compounds of formula (III) or (IV) represents any moiety suitable for covalently binding 2 or more GalNAc moieties, preferably 3 GalNac moieties, and the effector moiety linker LE.
- the bridging moiety B is a compound of formula (XV):
- the GalNAc linkers LGAL represent any chemical moiety suitable for covalently binding GalNAc to the bridging moiety as in formula (IV).
- the identity and size of the linker is not particularly limited.
- the GalNAc linkers LGAL each comprise 2-25 carbon atoms, preferably 7-15 carbon atoms, more preferably 11 carbon atoms.
- the GalNAc linkers LGAL comprise at least one, preferably two amide moieties.
- a particularly preferred GalNAc linker LGAL is a compound according to formula (XVI):
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule comprises or consists of at least one of a small molecule such as a drug molecule, a toxin such as a protein toxin, an oligonucleotide such as a BNA, a xeno nucleic acid or an siRNA, an enzyme, a peptide, a protein, or any combination thereof, preferably, the effector molecule is a toxin, an enzyme or an oligonucleotide.
- the effector molecule is a molecule capable of exerting a biological effect inside a cell that expresses the ASGPR on its surface and that comprises the binding partner for the effector molecule inside the cell, e.g. in the cytosol of said cell, such that the effector molecule can exert its intracellular biological effect once delivered inside the cell and preferably inside the cytosol, i.e. at the location inside the cell where the target for the effector molecule resides.
- An embodiment is the pharmaceutical combination of the invention, wherein the ligand for ASGPR, in particular a ligand for ASGPR1 , and the effector molecule, preferably a toxin or an oligonucleotide, are conjugated via a covalent bond, preferably via at least one linker.
- linkers suitable for coupling of the ligand for ASGPR, such as (GalNAc)3Tris, to an effector molecule, such as a BNA or an siRNA or a protein toxin, etc. are detailed here above.
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule is an oligonucleotide selected from any one or more of a(n): short interfering RNA (siRNA), short hairpin RNA (shRNA), anti-hairpin-shaped microRNA (miRNA), single-stranded RNA, aptamer RNA, double-stranded RNA (dsRNA), anti-microRNA (anti-miRNA, anti-miR), antisense oligonucleotide (ASO), mRNA, DNA, antisense DNA, locked nucleic acid (LNA), bridged nucleic acid (BNA), 2’-0,4’- aminoethylene bridged nucleic Acid (BNA NC ), BNA-based siRNA, and BNA-based antisense oligonucleotide (BNA-AON).
- siRNA short interfering RNA
- shRNA short hairpin RNA
- miRNA microRNA
- dsRNA double-stranded RNA
- any effector molecule for which the target binding partner is present inside a cell that expresses the receptor for the ligand comprised by the conjugate of the invention i.e. ASGPR
- ASGPR any effector molecule for which the target binding partner is present inside a cell that expresses the receptor for the ligand comprised by the conjugate of the invention, i.e. ASGPR
- an ASO, an siRNA or a BNA capable for targeting a gene, mRNA, etc., present in the cell which exposes ASGPR at its surface are suitable candidates for incorporation in the conjugate of the invention.
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule is an oligonucleotide selected from any one of: an anti-miRNA, a BNA-AON or an siRNA, such as BNA-based siRNA, preferably selected from chemically modified siRNA, metabolically stable siRNA and chemically modified, metabolically stable siRNA.
- an oligonucleotide selected from any one of: an anti-miRNA, a BNA-AON or an siRNA, such as BNA-based siRNA, preferably selected from chemically modified siRNA, metabolically stable siRNA and chemically modified, metabolically stable siRNA.
- Such chemically modified and/or metabolically stable siRNA moieties are known in the art, and are suitable for coupling to a ligand for ASGPR.
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule is an oligonucleotide that is capable of silencing any one of genes: HSP27, apolipoprotein B (apoB), transthyretin (TTR), proprotein convertase subtilisin/kexin type 9 (PCSK9), delta- aminolevulinate synthase 1 (ALAS1 ), antithrombin 3 (AT3), glycolate oxidase (GO), complement component C5 (CC5), X gene of hepatitis B virus (HBV), S gene of HBV, alpha-1 antitrypsin (AAT) and lactate dehydrogenase (LDH), and/or is capable of targeting an aberrant miRNA.
- HSP27 apolipoprotein B
- TTR transthyretin
- PCSK9 proprotein convertase subtilisin/kexin type 9
- PCSK9 proprotein convertase subtilisin/kexin type 9
- AAS1 delta-
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule, when present inside a cell, is an oligonucleotide that is capable of silencing any one of genes: apolipoprotein B (apoB), transthyretin (TTR), proprotein convertase subtilisin/kexin type 9 (PCSK9), delta-aminolevulinate synthase 1 (ALAS1 ), antithrombin 3 (AT3), glycolate oxidase (GO), complement component C5 (CC5), X gene of hepatitis B virus (HBV), S gene of HBV, alpha-1 antitrypsin (AAT) and lactate dehydrogenase (LDH), and/or wherein the effector molecule, when present inside a cell, is capable of targeting an aberrant miRNA.
- apoB apolipoprotein B
- TTR transthyretin
- PCSK9 proprotein convertase subtilisin/kexin type 9
- LAS1 delta
- any gene or mRNA present in a cell that expresses ASGPR at its cell surface can effectively be targeted by an effector molecule comprised by the conjugate of the invention, wherein the effector molecule is e.g. a BNA, an siRNA, an ASO, etc., when for example co administered together with a saponin according to the invention, to a subject in need of modification of e.g. a gene, (level of) mRNA expression, silencing of a gene, etc.
- an effector molecule comprised by the conjugate of the invention, wherein the effector molecule is e.g. a BNA, an siRNA, an ASO, etc.
- the therapeutic window of the conjugate comprising the effector molecule and the ligand for ASGPR is widened, resulting in a stronger biological effect of the effector molecule inside the target cell, and/or resulting in a biological effect of the effector molecule at a lower effective dose than what would be achieved when the target cells are exposed to the conjugate in the absence of the saponin or the saponin derivative.
- contacting the target cell which expresses ASGPR with a conjugate comprising a ligand for ASGPR and an effector molecule together with a saponin or a saponin derivative can result in overcoming a threshold with regard to intracellular activity of the effector molecule.
- a dose of the conjugate that for example would be required for reaching a sufficiently high amount of effector molecule in the cytosol of a target cell in the absence of saponin, could be too high for safe administration to a patient in need of such treatment with the effector molecule.
- the effector molecule already can exert its biological effect in the cytosol of a target cell expressing the ASGPR at lower dose, which lower dose would not exert any effect inside the cell when target cells are exposed to such lower dose in the absence of saponin.
- BNA for silencing HSP27 in (liver) cells did not silence HSP27 inside ASGPR bearing cells when a conjugate of GalNAc ligand for ASGPR and the BNA was contacted with ASGPR bearing cells, whereas when the same cells were contacted with the same or lower dose of the conjugate in the presence of saponin or saponin derivative such as a saponin with linker EMCH conjugated to the aldehyde group in the aglycone of the saponin, HSP27 was effectively silenced in the target cell (as shown in the appended example 2 and Figure 11 ).
- the inventors now provide molecules, pharmaceutical combinations, pharmaceutical compositions and a method for treating a (human) subject with such molecules, combinations and compositions, such that effector molecules can be administered to a patient in need thereof at lower dose, or such that effector molecules with an improved therapeutic window can be administered, resulting in an improved therapeutic effect once the effector molecule reaches its intracellular target in a target cell for the conjugates according to the invention.
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule is an oligonucleotide that, for example when present inside a mammalian cell, is capable of targeting an mRNA involved in expression of any one of proteins: HSP27, apoB, TTR, PCSK9, ALAS1 , AT3, GO, CC5, expression product of X gene of HBV, expression product of S gene of HBV, AAT and LDH, or is capable of antagonizing or restoring an miRNA function such as inhibiting an oncogenic miRNA (onco-miR) or suppression of expression of an onco-miR, for example when present inside a mammalian cell.
- proteins are typically involved in diseases for which e.g.
- RNAi enables target gene silencing by cleaving mRNA or repressing mRNA translation.
- the saponin aids in potentiating the gene silencing effector molecule by improving the cytosolic delivery of the effector molecule, when a target cell selected for RNAi therapy and expressing the ASGPR is contacted with both the saponin and the conjugate as described herein.
- an embodiment is the pharmaceutical combination of the invention, wherein the effector molecule is a toxin which comprises or consists of at least one proteinaceous molecule, preferably selected from any one or more of a peptide, a protein, an enzyme such as urease and Cre-recombinase, a proteinaceous toxin, a ribosome-inactivating protein, a protein toxin selected from Table A5 and/or a bacterial toxin, a plant toxin, more preferably selected from any one or more of a viral toxin such as apoptin; a bacterial toxin such as Shiga toxin, Shiga-like toxin, Pseudomonas aeruginosa exotoxin (PE) or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; a fungal toxin such as alpha-sarcin; a plant toxin including
- the protein toxin is dianthin and/or saporin, and/or comprises or consists of at least one of a toxin targeting ribosome, a toxin targeting elongation factor, a toxin targeting tubulin, a toxin targeting DNA and a toxin targeting RNA, more preferably any one or more of emtansine,
- the combination of the invention and the compositions of the invention are equally suitable for improved delivery of nucleic acid based effector molecules, when comprised by the conjugate of the ASGPR ligand and the effector molecule, as for cytosolic delivery of effector molecules such as peptides and proteins, when such an effector molecule as part of the conjugate is contacted with target cell expressing the ASGPR, in the presence of saponin or a saponin derivative.
- the therapeutic window for protein toxins is widened when contacting target cells with the conjugate comprising the effector molecule, i.e. the protein toxin, and the ligand for ASGPR, and with the saponin, according to the invention.
- the toxin exerts its intracellular effect to a higher extent or at a lower dose, when the target cells are exposed to the saponin and to the conjugate comprising the toxin, compared to the intracellular effect achieved at the same toxin dose or at the same lower toxin dose in the absence of the saponin, or a saponin derivative.
- An embodiment is the pharmaceutical combination of the invention, wherein the saponin comprises an aglycone core structure selected from the group (here referred to as ‘Group C’) consisting of: 2alpha-hydroxy oleanolic acid;
- the saponin comprises an aglycone core structure selected from quillaic acid and gypsogenin or derivatives thereof, more preferably the saponin aglycone core structure is quillaic acid or a derivative thereof.
- aglycone core structure selected from quillaic acid and gypsogenin or derivatives thereof, more preferably the saponin aglycone core structure is quillaic acid or a derivative thereof.
- Such preferred aglycones comprise an aldehyde group at the C-4 atom.
- such an aldehyde group contributes to the endosomal escape enhancing activity of saponins of the triterpene glycoside type, comprising an aglycone selected from Group C, especially quillajic acid and gypsogenin.
- An embodiment is the saponin conjugate of the invention, wherein the at least one saponin is a mono- desmosidic or bi-desmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with the aldehyde group in position C-4 and optionally comprising a glucuronic acid group in a carbohydrate substituent at the C-3beta-OH group of the saponin, preferably a bi-desmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with the aldehyde group in position C-4 and comprising a glucuronic acid group in a carbohydrate substituent at the C-3beta-OH group of the saponin.
- the at least one saponin is a mono- desmosidic or bi-desmosidic triterpene saponin belonging to the type of a 12,13-dehydrooleanane with the aldehyde group in position
- An embodiment is the pharmaceutical combination of the invention, wherein
- the saponin comprises a saccharide chain bound to the aglycone core structure, which is selected from group A:
- the saponin comprises a saccharide chain bound to the aglycone core structure, which is selected from group B:
- the saponin is a bidesmosidic triterpene glycoside comprising a first saccharide chain selected from the group A bound to the aglycone core structure and comprising a second saccharide chain selected from the group B bound to the aglycone core structure.
- Such saponins display the typical endosomal escape enhancing effect when a target cell is contacted with the saponin and an effector molecule, such as an effector molecule that is part of the conjugate according to the invention, comprising the effector molecule and the ligand for ASGPR1 , e.g. a (GalNAc)3Tris.
- an effector molecule such as an effector molecule that is part of the conjugate according to the invention, comprising the effector molecule and the ligand for ASGPR1 , e.g. a (GalNAc)3Tris.
- the effector molecule in the conjugate reaches an intracellular threshold level such that the biological activity of the effector molecule inside a target cell is either established, or is increased, or the activity of the effector molecule can be established at a lower dose than the dose required for the same level of activity in the absence of saponin.
- saponins with such endosomal escape enhancing activity are summarized. These saponins comprise the listed aglycone core structures of Group C and comprise the monosaccharides or polysaccharides of Group A and/or Group B, and have been shown to potentiate effector molecules when target cells are exposed to both the effector molecule and the saponin, or have high structural similarity with saponins for which such endosomal escape enhancing activity has been determined.
- an embodiment is the pharmaceutical combination of the invention, wherein the saponin is selected from the group consisting of: Quillaja bark saponin, dipsacoside B, saikosaponin A, saikosaponin D, macranthoidin A, esculentoside A, phytolaccagenin, aescinate, AS6.2, NP-005236, AMA-1 , AMR, alpha-Hederin, NP-012672, NP-017777, NP-017778, NP-017774, NP-018110, NP- 017772, NP-018109, NP-017888, NP-017889, NP-018108, SA1641 , AE X55, NP-017674, NP-017810, AG1 , NP-003881 , NP-017676, NP-017677, NP-017706, NP-017705, NP-017773, NP-017775, SA1657, AG2, S01861 , GE1741
- the present inventors have found that these derivatives have improved potentiating effects when employed in conjunction with a conjugate comprising an effector molecule as described herein. That is to say, the derivatives lead to comparable endosomal escape enhancing effects as the non-derivatised saponins, while displaying lower cytotoxicity and lower haemolytic activity. More structural details of saponin derivatives suitable in the context of the present invention are provided below.
- the saponin is a saponin derivative wherein either: i. the saponin derivative comprises an aglycone core structure comprising an aldehyde group which has been derivatised; or ii. the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group A as defined here above, the saccharide chain comprising a carboxyl group which has been derivatised; or iii. the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group B as defined here above, the saccharide chain comprising an acetoxy (Me(CO)O-) group which has been derivatised; or iv. any combination of derivatisations i., ii. and iii. is present, optionally any combination of two derivatisations of derivatisations i., ii. and iii.
- Also suitable according to the invention are the naturally occurring saponins for which the effector molecule potentiating effect is established when the saponin and the conjugate comprising the effector molecule are both contacted with a target cell exposing ASGPR1 at its surface.
- An embodiment is the pharmaceutical combination of the invention, wherein the saponin is selected from the group consisting of: S01861 , SA1657, GE1741 , SA1641 , QS-21 , QS-21 A, QS-21 A-api, QS- 21 A-xyl, QS-21 B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861 , QS1862, Quillajasaponin, Saponinum album, QS-18, Quil-A, Gyp1 , gypsoside A, AG1 , AG2, S01542, S01584, S01658, S01674, S01832, SO1904, stereoisomers thereof, derivatives thereof and combinations thereof, preferably the saponin is selected from the group consisting of QS-21 , a QS-21 derivative, S01861 , a S01861 derivative, SA16
- An embodiment is the pharmaceutical combination of the invention, wherein the saponin is a saponin derivative of the quillaic acid or gypsogenin saponin of Group C as listed here above and represented by Molecule 1 :
- Ai represents hydrogen, a monosaccharide or a linear or branched oligosaccharide, preferably Ai represents a saccharide chain selected from group A as defined here above, more preferably Ai represents a saccharide chain selected from group A as defined here above and Ai comprises or consists of a glucuronic acid moiety;
- A2 represents hydrogen, a monosaccharide or a linear or branched oligosaccharide, preferably A2 represents a saccharide chain selected from group B as defined here above, more preferably A2 represents a saccharide chain selected from group B as defined here above and A2 comprises at least one acetoxy (Me(CO)O-) group, such as one, two, three or four acetoxy groups, wherein at least one of Ai and A2 is not hydrogen, preferably both Ai and A2 are an oligosaccharide chain; and R is hydrogen in gypsogenin or hydroxyl in quillaic acid; wherein the saponin derivative corresponds to the saponin represented by Molecule 1 wherein at least one of the following derivatisations is present: i.
- the aldehyde group at position C230f the quillaic acid or gypsogenin has been derivatised; ii. the carboxyl group of a glucuronic acid moiety of Ai , when Ai represents a saccharide chain selected from group A as defined here above and Ai comprises or consists of a glucuronic acid moiety, has been derivatised; and iii. one or more, preferably all, of acetoxy group(s) of one saccharide moiety or of two or more saccharide moieties of A2, when A2 represents a saccharide chain selected from group B as defined here above and A2 comprises at least one acetoxy group, has/have been derivatised.
- An embodiment is the pharmaceutical combination of the invention, wherein Ai represents a saccharide chain selected from group A as defined here above and comprises or consists of a glucuronic acid moiety and wherein the carboxyl group of a glucuronic acid moiety of Ai has been derivatised and/or wherein A2 represents a saccharide chain selected from group B as defined here above and A2 comprises at least one acetoxy group and wherein at least one acetoxy group of A2 has been derivatised.
- An embodiment is the pharmaceutical combination of the invention, wherein the saponin represented by Molecule 1 is a bidesmosidic triterpene saponin, i.e. a saponin of the bidesmosidic triterpene glycoside type.
- An embodiment is the pharmaceutical combination of the invention, wherein the saponin derivative corresponds to the saponin represented by Molecule 1 wherein at least one of the following derivatisations is present: i. the aldehyde group at position C230f the quillaic acid or gypsogenin has been derivatised by;
- EMCH N-e-maleimidocaproic acid hydrazide
- BMPH N-[3-maleimidopropionic acid] hydrazide
- KMUH N-[K-maleimidoundecanoic acid] hydrazide
- the carboxyl group of a glucuronic acid moiety of Ai when Ai represents a saccharide chain selected from group A as defined here above and Ai comprises or consists of a glucuronic acid moiety, has been derivatised by transformation into an amide bond through reaction with 2-amino-2-methyl-1 ,3-propanediol (AMPD) or A/-(2-aminoethyl)maleimide (AEM), therewith providing a saponin-Glu-AMPD such as a QS-21-Glu-AMPD or a S01861-Glu- AMPD or a saponin-Glu-AEM such as a QS-21-Glu-AEM or a S01861-Glu-AEM; and iii.
- AMPD 2-amino-2-methyl-1 ,3-propanediol
- AEM A/-(2-aminoethyl)maleimide
- A2 represents a saccharide chain selected from group B as defined here above and A2 comprises at least one acetoxy group, has/have been derivatised by transformation into a hydroxyl group (HO-) by deacetylation.
- An embodiment is the pharmaceutical combination of the invention, wherein Ai is Gal-(1 - 2)- [Xyl-(1 - 3)]-GlcA and/or A 2 is Glc-(1 - 3)-Xyl-(1 - 4)-Rha-(1 - 2)-[Xyl-(1 - 3)-4-OAc-Qui-(1 - 4)]-Fuc, preferably the saponin represented by Molecule 1 is 3-0-beta-D-galactopyranosyl-(1 - 2)-[beta-D- xylopyranosyl-(1 - 3)]-beta-D-glucuronopyranosyl quillaic acid 28-0-beta-D-glucopyranosyl-(1 - 3)- beta-D-xylopyranosyl-(1 - 4)- alpha-L-rhamnopyranosyl-(1
- an embodiment is the pharmaceutical combination of the invention, wherein the saponin is a saponin derivative wherein either: i. the saponin derivative comprises an aglycone core structure comprising an aldehyde group which has been derivatised by: - reduction to an alcohol; or
- EMCH N-e-maleimidocaproic acid hydrazide
- BMPH N-[3-maleimidopropionic acid] hydrazide
- KMUH N-[K-maleimidoundecanoic acid] hydrazide
- the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group A as defined here above, the saccharide chain comprising a carboxyl group, preferably a carboxyl group of a glucuronic acid moiety which has been derivatised by transformation into an amide bond through reaction with 2-amino-2-methyl-1 ,3-propanediol (AMPD) or A/-(2-aminoethyl)maleimide (AEM), therewith providing a saponin-Glu-AMPD such as a QS-21-Glu- AMPD or a S01861-Glu-AMPD or a saponin-Glu-AEM such as a QS-21 -Glu- AEM or a SOI 861 -Glu-AEM; or iii.
- AMPD 2-amino-2-methyl-1 ,3-propanediol
- AEM A/-(2-aminoethyl)maleimide
- the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group B as defined here above, the saccharide chain comprising an acetoxy (Me(CO)O-) group which has been derivatised by transformation into a hydroxyl group (HO-) by deacetylation; or iv. the saponin derivative comprises any combination of derivatisations L, ii. and iii., optionally any combination of two derivatisations L, ii.
- the saponin derivative comprises an aglycone core structure wherein the aglycone core structure comprises an aldehyde group which has been derivatised by transformation into a hydrazone bond through reaction with EMCH wherein the maleimide group of the EMCH is optionally derivatised by formation of a thioether bond with mercaptoethanol.
- the saponin is a saponin derivative wherein either: i. the saponin derivative comprises an aglycone core structure comprising an aldehyde group which has been derivatised by transformation into a hydrazone bond through reaction with N- e-maleimidocaproic acid hydrazide (EMCH), therewith providing a saponin-Ald-EMCH such as a SOI 861 -Ald-EMCH or a QS-21 -Ald-EMCH; or ii.
- EMCH N- e-maleimidocaproic acid hydrazide
- the saponin comprises a saccharide chain, preferably a saccharide chain selected from group A as defined here above, the saccharide chain comprising a carboxyl group, preferably a carboxyl group of a glucuronic acid moiety which has been derivatised by transformation into an amide bond through reaction with A/-(2-aminoethyl)maleimide (AEM) , therewith providing a saponin-Glu-AEM such as a QS-21 -Glu-AEM or a SOI 861 -Glu-AEM; or iii. the saponin derivative comprises a combination of derivatisations i. and ii.
- AEM A/-(2-aminoethyl)maleimide
- an embodiment is the pharmaceutical combination of the invention, wherein the saponin derivative comprises an aglycone core structure wherein the aglycone core structure comprises an aldehyde group and wherein the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group A as defined here above, the saccharide chain comprising a carboxyl group, preferably a carboxyl group of a glucuronic acid moiety, which glucuronic acid moiety has been derivatised by transformation into an amide bond through reaction with A/-(2-aminoethyl)maleimide (AEM).
- AEM A/-(2-aminoethyl)maleimide
- An embodiment is the pharmaceutical combination of the invention, wherein the saponin is a saponin derivative represented by Molecule 2: or wherein the saponin derivative is the saponin derivative represented by Molecule 3
- a second aspect of the invention relates to the pharmaceutical combination of the invention for use as a medicament.
- the pharmaceutical combination of the invention for improving the efficacy of e.g. siRNA or BNA based therapies.
- Use of the pharmaceutical combination of the invention potentiates the intracellular effect of the effector molecule comprised by the conjugate comprising the ligand for ASGPR1 .
- the medicament is particularly suitable for treatment of diseases or abberancies involving aberrant liver cells which express ASGPR1 .
- Aberrant liver cells are cells of patients who suffer for example from any disease or health problem in which an expression product is involved of any one or more of genes: HSP27, apoB, TTR, PCSK9, ALAS1 , AT3, GO, CC5, X gene of HBV, S gene of HBV, AAT and LDH.
- Such diseases are for example any of a cancer, an infectious disease, a viral infection, hypercholesterolemia, primary hyperoxaluria, haemophilia A, haemophilia B, AAT related liver disease, acute hepatic porphyria, TTR-mediated amyloidosis, hereditary TTR amyloidosis (hATTR), complement-mediated disease, hepatitis B infection, a disease or disorder relating to HSP27 expression, or an auto-immune disease.
- a third aspect of the invention relates to the pharmaceutical combination of the invention, for use in the treatment or prophylaxis of a disease or health problem in which an expression product is involved of any one or more of genes: HSP27, apoB, TTR, PCSK9, ALAS1 , AT3, GO, CC5, X gene of HBV, S gene of HBV, AAT and LDH.
- An embodiment is the pharmaceutical combination of the invention, or the pharmaceutical combination for use of the invention, for use in the treatment or prophylaxis of a cancer, an infectious disease, a viral infection, hypercholesterolemia, primary hyperoxaluria, haemophilia A, haemophilia B, AAT related liver disease, acute hepatic porphyria, TTR-mediated amyloidosis, hereditary TTR amyloidosis (hATTR), complement-mediated disease, hepatitis B infection, a disease or disorder relating to HSP27 expression, or an auto-immune disease.
- An embodiment is the pharmaceutical combination for use according to the invention, wherein the pharmaceutical combination comprises:
- a saponin derivative wherein the saponin derivative corresponds to the saponin represented by Molecule 1 wherein at least one of the following derivatisations is present: i. the aldehyde group at position C230f the quillaic acid or gypsogenin has been derivatised by;
- EMCH N-e-maleimidocaproic acid hydrazide
- BMPH N-[3-maleimidopropionic acid] hydrazide
- KMUH N-[K-maleimidoundecanoic acid] hydrazide
- the carboxyl group of a glucuronic acid moiety of Ai when Ai represents a saccharide chain selected from group A and Ai comprises or consists of a glucuronic acid moiety, has been derivatised by transformation into an amide bond through reaction with 2-amino-2-methyl- 1 ,3-propanediol (AMPD) or A/-(2-aminoethyl)maleimide (AEM), therewith providing a saponin-Glu-AMPD such as a QS-21 -Glu-AMPD or a SOI 861 -Glu-AMPD or a saponin-Glu- AEM such as a QS-21 -Glu-AEM or a S01861 -Glu-AEM; and iii.
- AMPD 2-amino-2-methyl- 1 ,3-propanediol
- AEM A/-(2-aminoethyl)maleimide
- acetoxy group(s) of one saccharide moiety or of two or more saccharide moieties of A2 when A2 represents a saccharide chain selected from group B and A2 comprises at least one acetoxy group, has/have been derivatised by transformation into a hydroxyl group (HO-) by deacetylation; or
- the saponin represented by Molecule 1 is 3-0-beta-D-galactopyranosyl-(1 - 2)-[beta-D-xylopyranosyl- (1 - 3)]-beta-D-glucuronopyranosyl quillaic acid 28-0-beta-D-glucopyranosyl-(1 - 3)-beta-D- xylopyranosyl-(1 - 4)- alpha-L-rhamnopyranosy
- a saponin derivative wherein i. the saponin derivative comprises an aglycone core structure comprising an aldehyde group which has been derivatised by: - reduction to an alcohol; or
- EMCH N-e-maleimidocaproic acid hydrazide
- BMPH N-[3-maleimidopropionic acid] hydrazide
- KMUH N-[K-maleimidoundecanoic acid] hydrazide
- the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group A, the saccharide chain comprising a carboxyl group, preferably a carboxyl group of a glucuronic acid moiety which has been derivatised by transformation into an amide bond through reaction with 2-amino-2-methyl-1 ,3-propanediol (AMPD) or A/-(2-aminoethyl)maleimide (AEM), therewith providing a saponin-Glu-AMPD such as a QS-21-Glu- AMPD or a S01861- Glu-AMPD or a saponin-Glu-AEM such as a QS-21-Glu-AEM or a S01861-Glu-AEM; iii.
- AMPD 2-amino-2-methyl-1 ,3-propanediol
- AEM A/-(2-aminoethyl)maleimide
- the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group B, the saccharide chain comprising an acetoxy (Me(CO)O-) group which has been derivatised by transformation into a hydroxyl group (HO-) by deacetylation; or iv.
- the saponin derivative comprises any combination of derivatisations L, ii. and iii., optionally any combination of two derivatisations L, ii.
- the saponin derivative comprises an aglycone core structure wherein the aglycone core structure comprises an aldehyde group which has been derivatised by transformation into a hydrazone bond through reaction with EMCH wherein the maleimide group of the EMCH is optionally derivatised by formation of a thioether bond with mercaptoethanol; or
- a saponin derivative wherein i. the saponin derivative comprises an aglycone core structure comprising an aldehyde group which has been derivatised by transformation into a hydrazone bond through reaction with N- e-maleimidocaproic acid hydrazide (EMCH), therewith providing a saponin-Ald-EMCH such as a SOI 861 -Ald-EMCH or a QS-21 -Ald-EMCH; ii.
- EMCH N- e-maleimidocaproic acid hydrazide
- the saponin comprises a saccharide chain, preferably a saccharide chain selected from group A, the saccharide chain comprising a carboxyl group, preferably a carboxyl group of a glucuronic acid moiety which has been derivatised by transformation into an amide bond through reaction with A/-(2-aminoethyl)maleimide (AEM) , therewith providing a saponin-Glu- AEM such as a QS-21 -Glu-AEM or a SOI 861 -Glu-AEM; or iii. the saponin derivative comprises a combination of derivatisations i. and ii.; or
- a saponin derivative that comprises an aglycone core structure wherein the aglycone core structure comprises an aldehyde group and wherein the saponin derivative comprises a saccharide chain, preferably a saccharide chain selected from group A, the saccharide chain comprising a carboxyl group, preferably a carboxyl group of a glucuronic acid moiety, which glucuronic acid moiety has been derivatised by transformation into an amide bond through reaction with A/-(2-aminoethyl)maleimide (AEM); or • a saponin derivative represented by Molecule 2: or a saponin derivative represented by Molecule 3:
- a fourth aspect of the invention relates to an in vitro or ex vivo method for transferring the effector molecule comprised by the conjugate comprising the ligand for ASGPR according to the invention, from outside a cell to inside said cell, preferably into the cytosol of said cell, comprising the steps of: a) providing a cell which expresses ASGPR, preferably ASGPR1 , on its surface, preferably selected from a liver cell, a virally infected cell and a cancer cell; b) providing the conjugate of the invention, the conjugate comprising the effector molecule to be transferred; c) providing the saponin according to the invention; d) contacting the cell of step a) in vitro or ex vivo with the conjugate of step b) and the saponin of step c), therewith effecting the transfer of said conjugate comprising the effector molecule from outside the cell to inside said cell, and by effecting the transfer of said conjugate effecting the transfer of
- a fifth aspect of the invention relates to an in vitro or ex vivo method for transferring the conjugate of the invention from outside a cell to inside said cell, comprising the steps of: a) providing a cell which expresses ASGPR, preferably ASGPR1 , on its surface, preferably selected from a liver cell, a virally infected cell and a cancer cell; b) providing the conjugate of the invention; c) providing the saponin of the invention; d) contacting the cell of step a) in vitro or ex vivo with the conjugate of step b) and the saponin of step c), therewith effecting the transfer of the conjugate from outside the cell to inside said cell.
- an embodiment is any of the two methods of the invention, wherein the ligand for ASGPR comprises at least one /V-acetylgalactosamine (GalNAc) moiety, preferably three or four GalNAc moieties, more preferably the ligand for ASGPR comprises or consists of (GalNAc)3Tris and/or wherein the effector molecule is an oligonucleotide selected from any one of an anti-miRNA, a BNA-AON or an siRNA, such as BNA-based siRNA, preferably selected from chemically modified siRNA, metabolically stable siRNA and chemically modified, metabolically stable siRNA.
- GalNAc V-acetylgalactosamine
- an embodiment is any of the two methods of the invention, wherein the effector molecule comprises or consists of at least one of a small molecule such as a drug molecule, a toxin such as a protein toxin, an oligonucleotide such as a BNA, a xeno nucleic acid or an siRNA, an enzyme, a peptide, a protein, or any combination thereof, preferably, the effector molecule is a toxin, an enzyme or an oligonucleotide, preferably the toxin is saporin or dianthin.
- An embodiment is any of the two methods of the invention, wherein the saponin is derivatised SOI 861 and/or derivatised QS-21 , preferably SOI 861 -Glu-AEM or SOI 861 -Ald-EMCH or QS-21 -Glu- AEM or QS-21 -Ald-EMCH according to previous embodiments of the invention.
- An embodiment is any of the two methods of the invention, wherein the saponin is a saponin derivative corresponding to the saponin represented by Molecule 1 wherein at least one of the following derivatisations is present: i. the aldehyde group at position C230f the quillaic acid or gypsogenin has been derivatised by; - reduction to an alcohol; or
- EMCH N-e-maleimidocaproic acid hydrazide
- BMPH N-[3-maleimidopropionic acid] hydrazide
- KMUH N-[K-maleimidoundecanoic acid] hydrazide
- the carboxyl group of a glucuronic acid moiety of Ai when Ai represents a saccharide chain selected from group A and Ai comprises or consists of a glucuronic acid moiety, has been derivatised by transformation into an amide bond through reaction with 2-amino-2-methyl- 1 ,3-propanediol (AMPD) or A/-(2-aminoethyl)maleimide (AEM), therewith providing a saponin-Glu-AMPD such as a QS-21 -Glu-AMPD or a SOI 861 -Glu-AMPD or a saponin-Glu- AEM such as a QS-21 -Glu-AEM or a S01861 -Glu-AEM; and iii.
- AMPD 2-amino-2-methyl- 1 ,3-propanediol
- AEM A/-(2-aminoethyl)maleimide
- A2 represents a saccharide chain selected from group B and A2 comprises at least one acetoxy group, has/have been derivatised by transformation into a hydroxyl group (HO-) by deacetylation.
- HO- hydroxyl group
- an embodiment is any of the two methods of the invention, wherein the effector molecule is an oligonucleotide which, for example when present inside a mammalian cell, can silence any one of genes: apolipoprotein B (apoB), transthyretin (TTR), proprotein convertase subtilisin/kexin type 9 (PCSK9), delta-aminolevulinate synthase 1 (ALAS1 ), antithrombin 3 (AT3), glycolate oxidase (GO), complement component C5 (CC5), X gene of hepatitis B virus (HBV), S gene of HBV, alpha-1 antitrypsin (AAT) and lactate dehydrogenase (LDH), and/or, for example when present inside a mammalian cell, can target an aberrant miRNA and/or, for example when present inside a mammalian cell, wherein the oligonucleotide can target an mRNA involved in expression of any one of proteins: apoB,
- a sixth aspect of the invention relates to a kit of parts, comprising the pharmaceutical combination of the invention or the second pharmaceutical composition of the invention, and instructions for use of said pharmaceutical combination or second pharmaceutical composition according to the invention or for use in a method according to the invention.
- Trivalent-GalNAc is a targeting ligand that recognizes and binds the ASGPR1 receptor on hepatocytes.
- S01861 -EMCH refers to S01861 wherein the aldehyde group is derivatised by transformation into a hydrazone bond through reaction with N-e- maleimidocaproic acid hydrazide (EMCH), therewith providing S01861 -Ald-EMCH.
- the ‘trivalent- GalNAc’ as depicted in Figures 1 and 8 is a typical (GalNAc)3Tris conjugate suitable for coupling to an effector molecule or to saponin.
- HepG2 (ASGPR1 + ; CD71 + , Table A3) and Huh7 (ASGPR1 +/_ ; CD71 + , Table A3) cells were treated with a concentration range of trivalent-GalNAc-L-S01861 and GalNAc-L-S01861 in the presence and absence of 10 pM CD71 -saporin (monoclonal antibody OKT-9 targeting CD71 conjugated to the protein toxin saporin).
- Cell treatment in absence of CD71 -saporin show that the trivalent-GalNAc-L-S01861 (or trivalent-GalNAc, also referred to as (GalNAc)3) as single compound is not toxic up to 15000 nM ( Figure 9).
- CD71 -saporin also referred to as CD71 -SPRN
- DAR 1 for the bound SOI 861
- Targeted protein toxin mediated cell killing of ASGPR1/CD71 expressing cells was determined.
- trivalent-GalNAc-S01861 in combination with low concentrations of CD71 - saporin effectively induce cell killing in ASGPR1/CD71 expressing cells.
- trivalent-GalNAc-S01861 effectively induces endosomal escape of a protein toxin in ASGPR1 expressing cells.
- Example 2 trivalent-GalNAc-L/S-BNA + S01861-EMCH HSP27BNA was conjugated to trivalent-GalNAc ( Figure 6-7) and FlepG2 (ASGPR1 + ) cells or Fluh7 (ASPGR1 +/ ) were treated with a range of concentrations of trivalent-GalNAc-L-FISP27BNA (labile conjugation, Figure 6) or trivalent-GalNAc-S-FISP27BNA (stable conjugation, Figure 7) in combination with a fixed concentration of 4000 nM S01861 -EMCFI, also referred to as SPT-EMCFI.
- FISP27 gene silencing in FlepG2 and Fluh7 cells was determined.
- ApoBBNA was conjugated in a similar manner ( Figure 6-7) to trivalent-GalNAc and FlepG2 (ASGPR1 + ) cells or Fluh7 (ASPGR1 +/ ) were treated with a range of concentrations of trivalent-GalNAc- L-ApoBBNA (labile conjugation, Figure 6, Figure 12, Figure 13) or trivalent GalNAc-S-ApoBBNA (stable conjugation, Figure 7, Figure 14, Figure 15) or a trivalent GalNAc-L-ApoBscrambiedBNA or trivalent GalNAc-S-ApoBscrambiedBNA (where the scrambled ApoBBNA is an antisense scrambled oligo sequence that does not bind the ApoB mRNA; off-target control BNA) in combination with a fixed concentration of 4000 nM SOI 861 -EMCFI.
- the treatments did not affect the viability of the cell lines ( Figure 13, Figure 15).
- the ‘L’ refers to a ‘labile’ linker, which indicates that the linker is cleaved in the cell, i.e. at pH as apparent in the endosome and the lysosome.
- the ‘S’ refers to a ‘stable’ linker, which indicates that the linker is not cleaved in the cell, i.e. at pH as apparent in the endosome and the lysosome.
- a conjugate comprising a labile bond between any two molecules forming the conjugate or comprised by the conjugate, will be cleaved at a position in the labile linker, therewith dissociating the two linked or bound molecules.
- An example is the cleavage of a hydrazone bond under acidic conditions as apparent in the endosome and lysosome of mammalian cells such as human cells.
- S01861 -EMCH can enhance endosomal escape and target gene silencing of trivalent-GalNAc-L-BNA or trivalent-GalNAc-S-BNA for two independent gene targets.
- the gene silencing is more efficient in HepG2 cells compared to Huh7 cells, suggesting that higher levels of ASGPR1 expression at the plasma membrane of the cell facilitates increased uptake of the GalNAc- BNA conjugates.
- ApoBBNA (ApoBBNA#01 , targeting human ApoB mRNA) was conjugated to trivalent-GalNAc ((GalNAc)3) ( Figure 6) and primary hepatocytes (ASGPR1 + ) cells were treated with a range of concentrations of (GalNAc)3-ApoBBNA#01 with and without addition of 2000 nM S01861 . ApoB gene silencing in primary hepatocytes (ASGPR1 + ) cells was determined.
- Ls linker
- Apparatus Waters ICIass; Bin. Pump: UPIBSM, SM: UPISMFTN with SO; UPCMA, PDA: UPPDATC, 210-320 nm, SQD: ACQ-SQD2 ESI, mass ranges depending on the molecular weight of the product: neg or neg/pos within in a range of 1500-2400 or 2000-3000; ELSD: gaspressure 40 psi, drift tube temp: 50°C; column: Acquity C18, 50x2.1 mm, 1 .7 pm Temp: 60 S C, Flow: 0.6 mL/min, lin. Gradient depending on the polarity of the product:
- MS instrument type Agilent Technologies G6130B Quadrupole
- HPLC instrument type Agilent Technologies 1290 preparative LC
- S01861-EMCH synthesis (S01861-EMCH is also referred to as S01861-Ald-EMCH)
- MALDI-TOF-MS (RP mode): m/z 2193 Da ([M+K] + , S01861 -EMCH-mercaptoethanol), m/z 2185 Da ([M+K] + , S01861 -EMCH-mercaptoethanol), m/z 2170 Da ([M+Na] + , S01861 -EMCH-mercaptoethanol).
- reaction mixture was subjected to preparative MP-LC. 1 B Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (16.2 mg, 48%) as a white solid. Purity based on LC-MS 91 %.
- Trivalent GalNAc-azide (20.3 mg, 12.0 pmol) and 4- ⁇ 2-azatricyclo[10.4.0.0 49 ]hexadeca- 1 (12), 4(9), 5, 7,13, 15-hexaen-10-yn-2-yl ⁇ -N-[2-(2- ⁇ 2-[2-( ⁇ 4-[(E)- ⁇ [6-(2,5-dioxo-2,5-dihydro-1 H-pyrrol-1 - yl)hexanamido]imino ⁇ methyl]phenyl ⁇ formamido)ethoxy]ethoxy ⁇ ethoxy)ethyl]-4-oxobutanamide (11 .8 mg, 14.4 pmol) were dissolved in a mixture of water/acetonitrile (2:1 , v/v, 0.90 mL).
- Trivalent GalNAc-azide (20.3 mg, 12.0 mitioI) and DBCO-maleimide (10.3 mg, 24.0 mitioI) were dissolved in a mixture of water/acetonitrile (2:1 , v/v, 0.90 ml_). The reaction mixture was shaken for 1 min and left standing at room temperature. After 2 hours the reaction mixture was subjected to preparative MP-LC. 2D Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (22.2 mg, 87%) as a white solid. Purity based on LC-MS 85%. Contains 10% of the hydrolysed maleimide.
- the residue solution was diluted with 20 mM ammonium bicarbonate/acetonitrile (3:1 , v/v, 1.00 mL) and the resulting solution was directly added to trivalent GalNAc-S-maleimide (3.02 mg, 1 .43 pmol).
- the reaction mixture was shaken for 1 min and left standing at room temperature. After 1 .5 hours the reaction mixture was subjected to preparative LC-MS. 4A Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (6.75 mg, quant.) as a white fluffy solid. Purity based on LC-MS 94% (very broad peak).
- Trivalent GalNAc-S-ApoB (or HSP27) scrambled BNA oligo synthesis To ApoB scrambled BNA oligo disulfide (5.00 mg, 0.688 pmol) was added a solution of 20 mM ammonium bicarbonate with 2.5 mM TCEP (1 .00 mL, 2.5 pmol). The reaction mixture was shaken for 1 min and left standing at room temperature. After 1 hour the reaction mixture was filtered by using a centrifugal filter with a molecular weight cut-off of 3000 Da (5000 g for 30 min, 2 0.50 mL ).
- the residue solution was washed twice with a solution of 20 mM ammonium bicarbonate with 2.5 mM TCEP (0.50 mL), each time filtered under the same conditions described above.
- the residue solution was diluted with 20 mM ammonium bicarbonate/acetonitrile (3:1 , v/v, 1 .00 mL) and the resulting solution was directly added to trivalent GalNAc-S-maleimide (3.09 mg, 1 .46 pmol).
- the reaction mixture was shaken for 1 min and left standing at room temperature. After 1 .5 hours the reaction mixture was subjected to preparative LC-MS. 4A Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (5.91 mg, 93%) as a white fluffy solid. Purity based on LC-MS 88% (very broad peak).
- the residue solution was diluted with 20 mM ammonium bicarbonate/acetonitrile (3:1 , v/v, 1.00 ml_) and the resulting solution was directly added to trivalent GalNAc-L-maleimide (3.63 mg, 1 .45 pmol).
- the reaction mixture was shaken for 1 min and left standing at room temperature. After 1 .5 hours the reaction mixture was subjected to preparative LC-MS. 4A Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (6.68 mg, quant.) as a white fluffy solid. Purity based on LC-MS 99% (very broad peak).
- the residue solution was diluted with 20 mM ammonium bicarbonate/acetonitrile (3:1 , v/v, 1 .00 mL) and the resulting solution was directly added to trivalent GalNAc-L-maleimide (3.68 mg, 1 .47 prnol).
- the reaction mixture was shaken for 1 min and left standing at room temperature. After 1 .5 hours the reaction mixture was subjected to preparative LC-MS. 4A Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (4.71 mg, 71%) as a white fluffy solid. Purity based on LC-MS 96% (very broad peak).
- Trivalent GalNAc-azide (36.5 mg, 21 .6 prnol) was dissolved in a solution of potassium carbonate (5.97 mg, 43.2 prnol) in water (1 .00 mL) and acetonitrile (1 .00 mL).
- a 1 .0 M trimethylphosphine solution in THF (216 pL, 216 prnol) was added and the resulting mixture was shaken for 1 min and left standing at room temperature. After 45 min the reaction mixture was evaporated in vacuo and the residue was dissolved in water/acetonitrile (9:1 , v/v, 1 mL).
- the resulting solution was directly subjected to preparative MP-LC. 2B Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (36.1 mg, 98%) as a white solid. Purity based on LC-MS 100%.
- Trivalent GalNAc-amine formate (17.4 mg, 10.2 prnol) and DBCO-NHS (6.14 mg, 15.3 prnol) were dissolved in a solution of NMM (2.24 pL, 20.3 pmol) in DMF (0.50 mL). The reaction mixture was shaken for 1 min and left standing at room temperature. After 2 hours the reaction mixture was evaporated in vacuo and the residue was dissolved in water/acetonitrile (8:2, v/v, 1 mL). The resulting solution was directly subjected to preparative MP-LC. 2C Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (14.2 mg, 72%) as a white solid. Purity based on LC-MS 96%.
- Dendron(-L-S01861 ) 8 -amine (19.6 mg, 1.08 mitioI) and 2,5-dioxopyrrolidin-1 -yl 1 -azido-3,6,9,12- tetraoxapentadecan-15-oate (4.17 mg, 10.8 pmol) were dissolved in DMF (1 .50 ml_).
- DIPEA (1 .87 mI_, 10.8 mmol
- the reaction mixture was subjected to preparative LC-MS. 4B Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (11 .7 mg, 59%) as a white fluffy solid. Purity based on LC-MS 92% (very broad peak).
- Dendron(-L-S01861 )4-azide (2.50 mg, 0.266 pmol) and trivalent GalNAc-DBCO (1.56 mg, 0.799 pmol) were dissolved in a mixture of water/acetonitrile (3:1 , v/v, 1.00 ml_). The reaction mixture was shaken for 1 min and left standing at room temperature. After 2 hours the reaction mixture was subjected to preparative LC-MS. 4B Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (2.74 mg, 91%) as a white fluffy solid. Purity based on LC-MS 86% (very broad peak).
- Dendron(-L-S01861 )8-azide (2.50 mg, 0.135 pmol) and trivalent GalNAc-DBCO (0.79 mg, 0.405 pmol) were dissolved in a mixture of water/acetonitrile (3:1 , v/v, 1.00 mL). The reaction mixture was shaken for 1 min and left standing at room temperature. After 2 hours the reaction mixture was subjected to preparative LC-MS. 4B Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (2.03 mg, 74%) as a white fluffy solid. Purity based on LC-MS 100% (very broad peak).
- Trivalent GalNAc-amine formate (18.7 mg, 11 .0 mitioI) and 4-nitrophenyl 3-(acetylthio)propanoate (5.90 mg, 21 .9 pmol) were dissolved in a solution of NMM (2.41 mI_, 21 .9 mitioI) in DMF (0.50 ml_). The reaction mixture was shaken for 1 min and left standing at room temperature. After 2 hours the reaction mixture was evaporated in vacuo and the residue was dissolved in a mixture of 0.1% formic acid in water and 0.1% formic acid in acetonitrile (9:1 , v/v, 1 ml_). The resulting solution was directly subjected to preparative MP-LC.
- Trivalent GalNAc-thioacetate 13.0 mg, 7.25 pmol was dissolved in methanol (0.50 mL). Next, a 1 M solution of sodium hydroxide (7.98 pL, 7.98 pmol) was added. The reaction mixture was shaken for 1 min and left standing at room temperature. After 30 min the reaction mixture was added to a freshly prepared solution of trifunctional linker (Ls-t) (i.e., is an example of a saponin moiety linker Ls) (7.39 mg, 6.13 pmol) in 20 mM ammonium bicarbonate/acetonitrile (3:1 , v/v, 2.00 mL).
- Ls-t trifunctional linker
- the trifunctional linker has the lUPAC name: 5,8,11 ,18,21 ,24,27-Heptaoxa-2, 14,30- triazatritriacontanoic acid, 14-[16-(11 ,12-didehydrodibenz[t>,/]azocin-5(6/-/)-yl)-13,16-dioxo-3,6,9-trioxa- 12-azahexadec-1 -yl]-33-(2,5-dihydro-2,5-dioxo-1 /-/-pyrrol-1 -yl)-15,31 -dioxo-, (1 F?,4£)-4-cycloocten-1 -yl ester.
- the trifunctional linker has the following formula (XVII):
- reaction mixture was shaken for 1 min and left standing at room temperature. After 30 min the reaction mixture was frozen and lyophilized overnight to yield the crude title product as a pink fluffy solid.
- To the crude product was added a solution of 20 mM ammonium bicarbonate (1 .50 mL) and the resulting suspension was filtered over a 0.45 pm syringe filter. The filtrate was lyophilized overnight to yield the title product (5.44 mg, quant) as a pink fluffy solid. Purity based on LC-MS 90% (very broad peak).
- reaction mixture was shaken for 1 min and left standing at room temperature. After 30 min the reaction mixture was frozen and lyophilized overnight to yield the crude title product as a pink fluffy solid.
- To the crude product was added a solution of 20 mM ammonium bicarbonate (1 .50 ml_) and the resulting suspension was filtered over a 0.45 pm syringe filter. The filtrate was lyophilized overnight to yield the title product (5.48 mg, quant) as a pink fluffy solid. Purity based on LC-MS 84% (very broad peak).
- Trivalent GalNAc-thioacetate (16.2 mg, 9.02 mitioI) was dissolved in methanol (500 mI_). Next, a 1 .00 M solution of sodium hydroxide (11 .0 mI_, 11 .0 mitioI) was added. The reaction mixture was shaken for 1 min and left standing at room temperature. After 30 min the reaction mixture was submitted to preparative MP-LC. 1A Fractions corresponding to the product were immediately pooled together, frozen and lyophilized overnight to give the title compound (13.1 mg, 83%) as a white solid. Purity based on LC-MS 98%.
- Trifunctional linker (Ls) (15.0 mg, 12.4 mitioI) was dissolved in acetonitrile (0.50 mL). Next, a solution of 20 mM ammonium bicarbonate (1 .50 mL) was added and the resulting solution was directly transferred to trivalent GalNAc-thiol (22.7 mg, 13.0 pmol). The reaction mixture was shaken for 1 min and left standing at room temperature. After 1 hour the reaction mixture was frozen and lyophilized overnight to yield the crude title product as a white solid. Purity based on LC-MS 84%.
- Custom CD71 mab-saporin conjugate was produced and purchased from Advanced Targeting Systems (San Diego, CA).
- CD71 antibody anti-CD71 , clone OKT-9, InVivoMab was purchased from BioXCell.
- HSP27 (5’-GGCacagccagtgGCG-3’) [SEQ ID NO: 1] (The antisense BNA (HSP27) was BNA, more specifically BNA NC , with oligo nucleic acid sequence 5’-GGCacagccagtgGCG-3’ according to Zhang et al.
- RNA from cells was isolated and analysed according to standard protocols (Biorad). qPCR primers that were used are indicated in Table A2.
- the cells were incubated for 72 hr at 37°C before the cell viability was determined by a MTS-assay, performed according to the manufacturer’s instruction (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega). Briefly, the MTS solution was diluted 20x in DMEM without phenol red (PAN-Biotech GmbH) supplemented with 10% FBS. The cells were washed once with 200 pL/PBS well, after which 100 pL diluted MTS solution was added/well. The plate was incubated for approximately 20-30 minutes at 37°C. Subsequently, the OD at 492 nm was measured on a Thermo Scientific Multiskan FC plate reader (Thermo Scientific).
- the background signal of ‘medium only' wells was subtracted from all other wells, before the cell viability percentage of treated/untreated cells was calculated, by dividing the background corrected signal of treated wells over the background corrected signal of the untreated wells (x 100).
- Cells were seeded in DMEM (PAN-Biotech GmbH) supplemented with 10% fetal bovine serum (PAN- Biotech GmbH) and 1% penicillin/streptomycin (PAN-Biotech GmbH), in T75 flasks at appropriate density for each cell-line and incubated for 72-96 hrs (5% CO2, 37°C), until a confluency of 90% was reached.
- the cells were trypsinized (TryplE Express, Gibco Thermo Scientific) to single cells, transferred to a 15 ml_ falcon tube, and centrifuged (1 ,400 rpm, 3 min). The supernatant was discarded while leaving the cell pellet submerged.
- PE anti-human CD71 (#334106, Biolegend ) was used to stain the transferrin receptor, PE Mouse lgG2a, k Isotype Ctrl FC (#400212, Biolegend) was used as its matched isotype control.
- PE anti-human ASGPR1 (#130-122- 963, Miltenyi) was used to stain the ASGPR1 receptor and PE Mouse lgG1 , Isotype Ctrl (#130-113-762, Miltenyi) was used as its matched isotype control. Samples were incubated for 30 min. at 4°C.
- the cells were washed 2x with cold DPBS (Mg 2+ and Ca 2+ free, 2% FBS) and fixated for 20 min at room temperature using a 2% PFA solution in DPBS (Mg 2+ and Ca 2+ free, 2% FBS).
- Cells were washed 1x with cold DPBS, and resuspended in 1000 mI_ cold DPBS for FACS analysis. Samples were analyzed with a Sysmex Cube 8 flow cytometry system (Sysmex) and FCS Express 7 Research edition software. Results of the FACS analyses are summarized in Table A3.
- RNA from cells was isolated using TRI Reagent® Solution (Thermo Scientific) according to the manufacturer’s instruction. Conversion into cDNA was performed using iScriptTM cDNA Synthesis Kit (BioRad) using standard protocols. ApoB expression levels and levels of specific hepatocyte housekeeping genes were determined using quantitative real-time PCR assays (qRT-PCR) using iTaqTM Universal SYBR® Green Supermix (BioRad) and the Light Cycler 480 (Roche Diagnostics, Rotnch, Switzerland) with specific DNA primers, listed in Table A4 (murine) and Table A2 (human). Analysis was done by the ACt method to determine ApoB expression relative to 2 hepatocyte-specific housekeeping control mRNAs. Each analysis reaction was performed in triplicate.
- Cryopreserved primary mouse hepatocytes (PRIMACYT Cell Culture Technology GmbH, Germany) were thawed in hepatocyte thawing media (HTM, PRIMACYT Cell Culture Technology GmbH, Germany) and washed 1x with hepatocyte wash media (HWM, PRIMACYT Cell Culture Technology GmbH, Germany).
- Cells were re-suspended in hepatocyte plating media (HPM Cryo, PRIMACYT Cell Culture Technology GmbH, Germany) at a density of approx. 0.275 x 10 6 cells/ml.
- Cells were seeded onto collagen-l coated plates at a density of 88.000 cells/well or 26.400 cells/well for the 48 or 96-well plates (Greiner BioOne).
- Plating medium was replaced by 315 mI or 108 mI assay media (MHM, PRIMACYT Cell Culture Technology GmbH, Germany), after which conjugates were added from a 10x concentrated stock solution in PBS. Plates were incubated for 72 hr at 37°C being harvested for gene expression and cell viability analysis.
- Cryopreserved primary human hepatocytes (Cytes Biotechnologies S.L., Spain) were thawed in hepatocyte thawing media (Cytes Biotechnologies S.L., Spain). Cells were re-suspended in hepatocyte plating media (Cytes Biotechnologies S.L., Spain). Cells were seeded onto collagen-1 coated plates at a density of 215.600 cells/well or 66.600 cells/well for the 48 or 96-well plates (Greiner BioOne). Cells were pre-cultured for 4-6 hrs at 37°C allowing for cell attachment to cell culture plates before the start of treatment.
- Plating medium was replaced by 315 pi or 108 mI maintenance media (Cytes Biotechnologies S.L., Spain), after which conjugates were added from a 10x concentrated stock solution in PBS. Plates were incubated for 72 hr at 37°C being harvested for gene expression and cell viability analysis.
Abstract
Description
Claims
Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2022580238A JP2023533693A (en) | 2020-06-24 | 2021-06-23 | Therapeutic formulations of GalNAc-oligonucleotide conjugates and saponins, and uses thereof |
CN202180052532.8A CN116194148A (en) | 2020-06-24 | 2021-06-23 | Therapeutic combinations of GalNAc-oligonucleotide conjugates and saponins and uses thereof |
US18/012,778 US20230293562A1 (en) | 2020-06-24 | 2021-06-23 | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof |
IL299349A IL299349A (en) | 2020-06-24 | 2021-06-23 | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof |
KR1020237002644A KR20230044196A (en) | 2020-06-24 | 2021-06-23 | Therapeutic Combinations of GALNAC-oligonucleotide Conjugates and Saponins and Their Uses |
AU2021296983A AU2021296983A1 (en) | 2020-06-24 | 2021-06-23 | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof |
EP21734544.6A EP4171643A1 (en) | 2020-06-24 | 2021-06-23 | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof |
CA3183881A CA3183881A1 (en) | 2020-06-24 | 2021-06-23 | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
NL2025902 | 2020-06-24 | ||
NL2025902 | 2020-06-24 | ||
NLPCT/NL2021/050384 | 2021-06-18 | ||
PCT/NL2021/050384 WO2021261992A1 (en) | 2020-06-24 | 2021-06-18 | Conjugate of galnac and saponin, therapeutic composition comprising said conjugate and a galnac-oligonucleotide conjugate |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2021261998A1 true WO2021261998A1 (en) | 2021-12-30 |
Family
ID=76601666
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/NL2021/050394 WO2021261998A1 (en) | 2020-06-24 | 2021-06-23 | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof |
Country Status (9)
Country | Link |
---|---|
US (1) | US20230293562A1 (en) |
EP (1) | EP4171643A1 (en) |
JP (1) | JP2023533693A (en) |
KR (1) | KR20230044196A (en) |
CN (1) | CN116194148A (en) |
AU (1) | AU2021296983A1 (en) |
CA (1) | CA3183881A1 (en) |
IL (1) | IL299349A (en) |
WO (1) | WO2021261998A1 (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011054811A1 (en) * | 2009-11-03 | 2011-05-12 | Santaris Pharma A/S | Rna antagonists targeting hsp27 combination therapy |
WO2015071388A1 (en) * | 2013-11-14 | 2015-05-21 | Roche Innovation Center Copenhagen A/S | Apob antisense conjugate compounds |
WO2020126604A1 (en) * | 2018-12-21 | 2020-06-25 | Sapreme Technologies B.V. | Saponin conjugates |
-
2021
- 2021-06-23 AU AU2021296983A patent/AU2021296983A1/en active Pending
- 2021-06-23 KR KR1020237002644A patent/KR20230044196A/en unknown
- 2021-06-23 WO PCT/NL2021/050394 patent/WO2021261998A1/en active Search and Examination
- 2021-06-23 JP JP2022580238A patent/JP2023533693A/en active Pending
- 2021-06-23 EP EP21734544.6A patent/EP4171643A1/en active Pending
- 2021-06-23 IL IL299349A patent/IL299349A/en unknown
- 2021-06-23 CN CN202180052532.8A patent/CN116194148A/en active Pending
- 2021-06-23 CA CA3183881A patent/CA3183881A1/en active Pending
- 2021-06-23 US US18/012,778 patent/US20230293562A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011054811A1 (en) * | 2009-11-03 | 2011-05-12 | Santaris Pharma A/S | Rna antagonists targeting hsp27 combination therapy |
WO2015071388A1 (en) * | 2013-11-14 | 2015-05-21 | Roche Innovation Center Copenhagen A/S | Apob antisense conjugate compounds |
WO2020126604A1 (en) * | 2018-12-21 | 2020-06-25 | Sapreme Technologies B.V. | Saponin conjugates |
Non-Patent Citations (14)
Title |
---|
AARON D. SPRINGER ET AL: "GalNAc-siRNA Conjugates: Leading the Way for Delivery of RNAi Therapeutics", NUCLEIC ACID THERAPEUTICS, vol. 28, no. 3, 1 June 2018 (2018-06-01), US, pages 109 - 118, XP055555952, ISSN: 2159-3337, DOI: 10.1089/nat.2018.0736 * |
ALEXANDER WENG ET AL: "The toxin component of targeted anti-tumor toxins determines their efficacy increase by saponins", MOLECULAR ONCOLOGY, ELSEVIER, vol. 6, no. 3, 12 January 2012 (2012-01-12), pages 323 - 332, XP028508748, ISSN: 1574-7891, [retrieved on 20120124], DOI: 10.1016/J.MOLONC.2012.01.004 * |
CHEENU BHARGAVA ET AL: "Targeted dianthin is a powerful toxin to treat pancreatic carcinoma when applied in combination with the glycosylated triterpene SO1861", MOLECULAR ONCOLOGY, vol. 11, no. 11, 15 September 2017 (2017-09-15), pages 1527 - 1543, XP055677214, ISSN: 1574-7891, DOI: 10.1002/1878-0261.12115 * |
GILABERT-ORIOL ROGER ET AL: "Dianthin-30 or gelonin versus monomethyl auristatin E, each configured with an anti-calcitonin receptor antibody, are differentially potent in vitro in high-grade glioma cell lines derived from glioblastoma", CANCER IMMUNOLOGY, IMMUNOTHERAPY, SPRINGER, BERLIN/HEIDELBERG, vol. 66, no. 9, 13 May 2017 (2017-05-13), pages 1217 - 1228, XP036309505, ISSN: 0340-7004, [retrieved on 20170513], DOI: 10.1007/S00262-017-2013-Z * |
GOODING, MATT ET AL., CHEMICAL BIOLOGY & DRUG DESIGN, vol. 80.6, 2012, pages 787 - 809 |
HENDRIK FUCHS ET AL: "Glycosylated Triterpenoids as Endosomal Escape Enhancers in Targeted Tumor Therapies", BIOMEDICINES, vol. 5, no. 2, 29 March 2017 (2017-03-29), pages 14, XP055429235, DOI: 10.3390/biomedicines5020014 * |
HERMANSON, GREG T.: "Bioconjugate techniques", 2013, ACADEMIC PRESS, pages: 289 |
J. AM. CHEM SOC., vol. 136, 2014, pages 16958 - 16961 |
ROGER GILABERT-ORIOL ET AL: "Combinatorial approach to increase efficacy of Cetuximab, Panitumumab and Trastuzumab by dianthin conjugation and co-application of SO1861", BIOCHEMICAL PHARMACOLOGY, vol. 97, no. 3, 1 October 2015 (2015-10-01), US, pages 247 - 255, XP055677222, ISSN: 0006-2952, DOI: 10.1016/j.bcp.2015.07.040 * |
ROGER GILABERT-ORIOL ET AL: "Modified Trastuzumab and Cetuximab Mediate Efficient Toxin Delivery While Retaining Antibody-Dependent Cell-Mediated Cytotoxicity in Target Cells", MOLECULAR PHARMACEUTICS, vol. 10, no. 11, 7 October 2013 (2013-10-07), US, pages 4347 - 4357, XP055677139, ISSN: 1543-8384, DOI: 10.1021/mp400444q * |
SABINE SEWING ET AL: "GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells", MOLECULAR THERAPY: NUCLEIC ACIDS., vol. 14, 1 March 2019 (2019-03-01), US, pages 67 - 79, XP055762603, ISSN: 2162-2531, DOI: 10.1016/j.omtn.2018.11.005 * |
SETTEN, RYAN L.JOHN J. ROSSISI-PING HAN, NATURE REVIEWS DRUG DISCOVERY, vol. 18.6, 2019, pages 421 - 446 |
Y ZHANGZ QUS KIMV SHIB LIAO 1P KRAFTR BANDARUY WULM GREENBERGERID HORAK: "Down-modulation of cancer targets using locked nucleic acid (LNA)-based antisense oligonucleotides without transfection", GENE THERAPY, vol. 18, 2011, pages 326 - 333, XP055412231, DOI: 10.1038/gt.2010.133 |
YUANYU HUANG: "Preclinical and Clinical Advances of GalNAc-Decorated Nucleic Acid Therapeutics", MOLECULAR THERAPY: NUCLEIC ACIDS., vol. 6, 1 March 2017 (2017-03-01), US, pages 116 - 132, XP055485332, ISSN: 2162-2531, DOI: 10.1016/j.omtn.2016.12.003 * |
Also Published As
Publication number | Publication date |
---|---|
JP2023533693A (en) | 2023-08-04 |
CN116194148A (en) | 2023-05-30 |
KR20230044196A (en) | 2023-04-03 |
US20230293562A1 (en) | 2023-09-21 |
AU2021296983A1 (en) | 2023-02-23 |
EP4171643A1 (en) | 2023-05-03 |
IL299349A (en) | 2023-02-01 |
CA3183881A1 (en) | 2021-12-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6259448B2 (en) | Novel conjugates comprising TETRA GALNAC and peptides and methods for delivering oligonucleotides | |
WO2021261992A1 (en) | Conjugate of galnac and saponin, therapeutic composition comprising said conjugate and a galnac-oligonucleotide conjugate | |
US20230263897A1 (en) | Conjugate of galnac and saponin, therapeutic composition comprising said conjugate and a galnac-oligonucleotide conjugate | |
WO2022055352A1 (en) | Semicarbazone-based saponin conjugate | |
US20230357310A1 (en) | Saponin derivatives with improved therapeutic window | |
WO2022055351A1 (en) | Conjugate of saponin, oligonucleotide and galnac | |
US20230293562A1 (en) | Therapeutic combination of galnac-oligonucleotide conjugate and saponin, and uses thereof | |
US20230277612A1 (en) | Saponin derivatives for use in medicine | |
WO2022265493A1 (en) | Conjugate of saponin, oligonucleotide and galnac | |
AU2021339332A1 (en) | Conjugate of saponin, oligonucleotide and GalNAc | |
CN117881427A (en) | Conjugates of saponins, oligonucleotides and GALNAC | |
US20240066045A1 (en) | Semicarbazone-based saponin conjugate | |
WO2023038517A1 (en) | Semicarbazone-based saponin conjugate |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 21734544 Country of ref document: EP Kind code of ref document: A1 |
|
DPE1 | Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101) | ||
ENP | Entry into the national phase |
Ref document number: 3183881 Country of ref document: CA |
|
ENP | Entry into the national phase |
Ref document number: 2022580238 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2021734544 Country of ref document: EP Effective date: 20230124 |
|
ENP | Entry into the national phase |
Ref document number: 2021296983 Country of ref document: AU Date of ref document: 20210623 Kind code of ref document: A |