US20070021327A1 - Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor - Google Patents
Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor Download PDFInfo
- Publication number
- US20070021327A1 US20070021327A1 US11/518,321 US51832106A US2007021327A1 US 20070021327 A1 US20070021327 A1 US 20070021327A1 US 51832106 A US51832106 A US 51832106A US 2007021327 A1 US2007021327 A1 US 2007021327A1
- Authority
- US
- United States
- Prior art keywords
- pdgf
- nucleic acid
- nucleic acids
- selex
- tumors
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 239000003446 ligand Substances 0.000 title claims abstract description 92
- 238000000034 method Methods 0.000 title claims abstract description 77
- 239000000203 mixture Substances 0.000 title claims abstract description 58
- 108020004707 nucleic acids Proteins 0.000 title claims description 117
- 102000039446 nucleic acids Human genes 0.000 title claims description 117
- 150000007523 nucleic acids Chemical class 0.000 title claims description 117
- 206010028980 Neoplasm Diseases 0.000 title abstract description 83
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 title description 36
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 title description 36
- 238000011282 treatment Methods 0.000 title description 17
- 108091023037 Aptamer Proteins 0.000 claims abstract description 40
- 229940127089 cytotoxic agent Drugs 0.000 claims abstract description 19
- 239000002254 cytotoxic agent Substances 0.000 claims abstract description 18
- 231100000599 cytotoxic agent Toxicity 0.000 claims abstract description 18
- 230000001965 increasing effect Effects 0.000 claims abstract description 16
- 230000027455 binding Effects 0.000 claims description 16
- 238000009739 binding Methods 0.000 claims description 16
- 230000001225 therapeutic effect Effects 0.000 claims description 14
- 238000000638 solvent extraction Methods 0.000 claims description 11
- 229930012538 Paclitaxel Natural products 0.000 claims description 9
- 229960001592 paclitaxel Drugs 0.000 claims description 9
- 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 9
- 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 6
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 claims description 6
- -1 Doxofluoridine Chemical compound 0.000 claims description 4
- NDMPLJNOPCLANR-UHFFFAOYSA-N 3,4-dihydroxy-15-(4-hydroxy-18-methoxycarbonyl-5,18-seco-ibogamin-18-yl)-16-methoxy-1-methyl-6,7-didehydro-aspidospermidine-3-carboxylic acid methyl ester Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 NDMPLJNOPCLANR-UHFFFAOYSA-N 0.000 claims description 3
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin 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-VTZDEGQISA-N 0.000 claims description 3
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 claims description 3
- 108010006654 Bleomycin Proteins 0.000 claims description 3
- 190000008236 Carboplatin Chemical compound 0.000 claims description 3
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 claims description 3
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 claims description 3
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 claims description 3
- 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 3
- 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 3
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 claims description 3
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 claims description 3
- 229960001561 bleomycin Drugs 0.000 claims description 3
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical class N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 claims description 3
- 229960004562 carboplatin Drugs 0.000 claims description 3
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical class N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 3
- 229960004316 cisplatin Drugs 0.000 claims description 3
- 229960004397 cyclophosphamide Drugs 0.000 claims description 3
- 229960000975 daunorubicin Drugs 0.000 claims description 3
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin 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(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 claims description 3
- CFCUWKMKBJTWLW-UHFFFAOYSA-N deoliosyl-3C-alpha-L-digitoxosyl-MTM Natural products CC=1C(O)=C2C(O)=C3C(=O)C(OC4OC(C)C(O)C(OC5OC(C)C(O)C(OC6OC(C)C(O)C(C)(O)C6)C5)C4)C(C(OC)C(=O)C(O)C(C)O)CC3=CC2=CC=1OC(OC(C)C1O)CC1OC1CC(O)C(O)C(C)O1 CFCUWKMKBJTWLW-UHFFFAOYSA-N 0.000 claims description 3
- 229960004679 doxorubicin Drugs 0.000 claims description 3
- 229960001904 epirubicin Drugs 0.000 claims description 3
- 229960005420 etoposide Drugs 0.000 claims description 3
- 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 3
- 229960002949 fluorouracil Drugs 0.000 claims description 3
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 claims description 3
- 229940020967 gemzar Drugs 0.000 claims description 3
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 claims description 3
- 229960001101 ifosfamide Drugs 0.000 claims description 3
- 229950010897 iproplatin Drugs 0.000 claims description 3
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 claims description 3
- SGDBTWWWUNNDEQ-LBPRGKRZSA-N melphalan Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N(CCCl)CCCl)C=C1 SGDBTWWWUNNDEQ-LBPRGKRZSA-N 0.000 claims description 3
- 229960001924 melphalan Drugs 0.000 claims description 3
- 229960000485 methotrexate Drugs 0.000 claims description 3
- CFCUWKMKBJTWLW-BKHRDMLASA-N mithramycin Chemical compound O([C@@H]1C[C@@H](O[C@H](C)[C@H]1O)OC=1C=C2C=C3C[C@H]([C@@H](C(=O)C3=C(O)C2=C(O)C=1C)O[C@@H]1O[C@H](C)[C@@H](O)[C@H](O[C@@H]2O[C@H](C)[C@H](O)[C@H](O[C@@H]3O[C@H](C)[C@@H](O)[C@@](C)(O)C3)C2)C1)[C@H](OC)C(=O)[C@@H](O)[C@@H](C)O)[C@H]1C[C@@H](O)[C@H](O)[C@@H](C)O1 CFCUWKMKBJTWLW-BKHRDMLASA-N 0.000 claims description 3
- 229960004857 mitomycin Drugs 0.000 claims description 3
- 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 3
- 229960001156 mitoxantrone Drugs 0.000 claims description 3
- 229960003171 plicamycin Drugs 0.000 claims description 3
- 229960001052 streptozocin Drugs 0.000 claims description 3
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 claims description 3
- 229940063683 taxotere Drugs 0.000 claims description 3
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 claims description 3
- 229960000303 topotecan Drugs 0.000 claims description 3
- 229960003048 vinblastine Drugs 0.000 claims description 3
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 claims description 3
- 229960004528 vincristine Drugs 0.000 claims description 3
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 claims description 3
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 claims description 3
- 229960004355 vindesine Drugs 0.000 claims description 3
- UGGWPQSBPIFKDZ-KOTLKJBCSA-N vindesine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(N)=O)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1N=C1[C]2C=CC=C1 UGGWPQSBPIFKDZ-KOTLKJBCSA-N 0.000 claims description 3
- GBABOYUKABKIAF-GHYRFKGUSA-N vinorelbine Chemical compound C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC GBABOYUKABKIAF-GHYRFKGUSA-N 0.000 claims description 3
- 229960002066 vinorelbine Drugs 0.000 claims description 3
- UIYWFOZZIZEEKJ-XVFCMESISA-N 1-[(2r,3r,4r,5r)-3-fluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidine-2,4-dione Chemical compound F[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 UIYWFOZZIZEEKJ-XVFCMESISA-N 0.000 claims 1
- NVZFZMCNALTPBY-XVFCMESISA-N 4-amino-1-[(2r,3r,4r,5r)-3-fluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](F)[C@H](O)[C@@H](CO)O1 NVZFZMCNALTPBY-XVFCMESISA-N 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 18
- 230000009897 systematic effect Effects 0.000 abstract description 16
- 210000003722 extracellular fluid Anatomy 0.000 abstract description 5
- 230000004048 modification Effects 0.000 description 29
- 238000012986 modification Methods 0.000 description 29
- 108091034117 Oligonucleotide Proteins 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 15
- 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 14
- 108010081589 Becaplermin Proteins 0.000 description 11
- 239000003814 drug Substances 0.000 description 11
- 125000003729 nucleotide group Chemical group 0.000 description 11
- 241000700159 Rattus Species 0.000 description 10
- 229940079593 drug Drugs 0.000 description 9
- 210000004881 tumor cell Anatomy 0.000 description 9
- 230000003321 amplification Effects 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 238000003199 nucleic acid amplification method Methods 0.000 description 8
- 201000011510 cancer Diseases 0.000 description 7
- 239000003112 inhibitor Substances 0.000 description 7
- 108010000685 platelet-derived growth factor AB Proteins 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- 238000009472 formulation Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000006467 substitution reaction Methods 0.000 description 6
- 102000018967 Platelet-Derived Growth Factor beta Receptor Human genes 0.000 description 5
- 108010051742 Platelet-Derived Growth Factor beta Receptor Proteins 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 238000012384 transportation and delivery Methods 0.000 description 5
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 4
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 4
- 150000003230 pyrimidines Chemical group 0.000 description 4
- 238000007920 subcutaneous administration Methods 0.000 description 4
- ZFXYFBGIUFBOJW-UHFFFAOYSA-N theophylline Chemical compound O=C1N(C)C(=O)N(C)C2=C1NC=N2 ZFXYFBGIUFBOJW-UHFFFAOYSA-N 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 208000001446 Anaplastic Thyroid Carcinoma Diseases 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 108091008606 PDGF receptors Proteins 0.000 description 3
- 229910019142 PO4 Chemical group 0.000 description 3
- 102000011653 Platelet-Derived Growth Factor Receptors Human genes 0.000 description 3
- 238000011579 SCID mouse model Methods 0.000 description 3
- 238000000692 Student's t-test Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 239000010452 phosphate Chemical group 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 230000009870 specific binding Effects 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 201000008440 thyroid gland anaplastic carcinoma Diseases 0.000 description 3
- 208000019179 thyroid gland undifferentiated (anaplastic) carcinoma Diseases 0.000 description 3
- RGNOTKMIMZMNRX-XVFCMESISA-N 2-amino-1-[(2r,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-4-one Chemical compound NC1=NC(=O)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 RGNOTKMIMZMNRX-XVFCMESISA-N 0.000 description 2
- ZLOIGESWDJYCTF-UHFFFAOYSA-N 4-Thiouridine Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=S)C=C1 ZLOIGESWDJYCTF-UHFFFAOYSA-N 0.000 description 2
- ZLOIGESWDJYCTF-XVFCMESISA-N 4-thiouridine Chemical class O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=S)C=C1 ZLOIGESWDJYCTF-XVFCMESISA-N 0.000 description 2
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical class BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 2
- KSNXJLQDQOIRIP-UHFFFAOYSA-N 5-iodouracil Chemical class IC1=CNC(=O)NC1=O KSNXJLQDQOIRIP-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical group OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 2
- 108091008102 DNA aptamers Proteins 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 108700020121 Human Immunodeficiency Virus-1 rev Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 2
- 241001529936 Murinae Species 0.000 description 2
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 2
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 229960001948 caffeine Drugs 0.000 description 2
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 125000003636 chemical group Chemical group 0.000 description 2
- 238000007385 chemical modification Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000001351 cycling effect Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 238000006073 displacement reaction Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000009881 electrostatic interaction Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 230000000762 glandular Effects 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- 230000011987 methylation Effects 0.000 description 2
- 238000007069 methylation reaction Methods 0.000 description 2
- 230000000269 nucleophilic effect Effects 0.000 description 2
- 125000003835 nucleoside group Chemical group 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical group [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 229960000278 theophylline Drugs 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 0 *[U].C.C.C.C.C.C.CC.CC.CCOP(=O)([O-])OCCCCCNC(=O)C(CCCCNC(=O)OC)NC(=O)OC.[3HH].[3HH].[3HH].[U].[U] Chemical compound *[U].C.C.C.C.C.C.CC.CC.CCOP(=O)([O-])OCCCCCNC(=O)C(CCCCNC(=O)OC)NC(=O)OC.[3HH].[3HH].[3HH].[U].[U] 0.000 description 1
- HWTAKVLMACWHLD-UHFFFAOYSA-N 2-(9h-carbazol-1-yl)ethanamine Chemical compound C12=CC=CC=C2NC2=C1C=CC=C2CCN HWTAKVLMACWHLD-UHFFFAOYSA-N 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 102000006306 Antigen Receptors Human genes 0.000 description 1
- 108010083359 Antigen Receptors Proteins 0.000 description 1
- 238000011603 BDIX rat Methods 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- JVSUHLJQOVNHKD-UHFFFAOYSA-M COCCOCCOP(=O)([O-])OC Chemical compound COCCOCCOP(=O)([O-])OC JVSUHLJQOVNHKD-UHFFFAOYSA-M 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 206010010144 Completed suicide Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 102000001393 Platelet-Derived Growth Factor alpha Receptor Human genes 0.000 description 1
- 108010068588 Platelet-Derived Growth Factor alpha Receptor Proteins 0.000 description 1
- 102000004278 Receptor Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108090000873 Receptor Protein-Tyrosine Kinases Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 102000012740 beta Adrenergic Receptors Human genes 0.000 description 1
- 108010079452 beta Adrenergic Receptors Proteins 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000002051 biphasic effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000003028 elevating effect Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000002991 immunohistochemical analysis Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 238000010874 in vitro model Methods 0.000 description 1
- 238000005462 in vivo assay Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 150000002605 large molecules Chemical class 0.000 description 1
- 150000002634 lipophilic molecules Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 201000005296 lung carcinoma Diseases 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 239000003226 mitogen Substances 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000008177 pharmaceutical agent Substances 0.000 description 1
- 150000004713 phosphodiesters Chemical group 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 125000000714 pyrimidinyl group Chemical group 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 239000006215 rectal suppository Substances 0.000 description 1
- 229940100618 rectal suppository Drugs 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000002536 stromal cell Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012385 systemic delivery Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 1
- 239000003104 tissue culture media Substances 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 239000006216 vaginal suppository Substances 0.000 description 1
- 229940120293 vaginal suppository Drugs 0.000 description 1
- 210000005167 vascular cell Anatomy 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
- C40B50/04—Methods of creating libraries, e.g. combinatorial synthesis using dynamic combinatorial chemistry techniques
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
- A61K47/544—Phospholipids
-
- 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
- A61K47/547—Chelates, e.g. Gd-DOTA or Zinc-amino acid chelates; Chelate-forming compounds, e.g. DOTA or ethylenediamine being covalently linked or complexed to the pharmacologically- or therapeutically-active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/49—Platelet-derived growth factor [PDGF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/495—Transforming growth factor [TGF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/50—Fibroblast growth factor [FGF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1048—SELEX
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1136—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1276—RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6811—Selection methods for production or design of target specific oligonucleotides or binding molecules
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
- G01N33/56988—HIV or HTLV
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6842—Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/13—Decoys
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/31—Chemical structure of the backbone
- C12N2310/317—Chemical structure of the backbone with an inverted bond, e.g. a cap structure
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/31—Chemical structure of the backbone
- C12N2310/318—Chemical structure of the backbone where the PO2 is completely replaced, e.g. MMI or formacetal
- C12N2310/3183—Diol linkers, e.g. glycols or propanediols
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/32—Chemical structure of the sugar
- C12N2310/321—2'-O-R Modification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/32—Chemical structure of the sugar
- C12N2310/322—2'-R Modification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
- C12N2310/3515—Lipophilic moiety, e.g. cholesterol
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/50—Physical structure
- C12N2310/53—Physical structure partially self-complementary or closed
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
-
- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F02—COMBUSTION ENGINES; HOT-GAS OR COMBUSTION-PRODUCT ENGINE PLANTS
- F02B—INTERNAL-COMBUSTION PISTON ENGINES; COMBUSTION ENGINES IN GENERAL
- F02B75/00—Other engines
- F02B75/02—Engines characterised by their cycles, e.g. six-stroke
- F02B2075/022—Engines characterised by their cycles, e.g. six-stroke having less than six strokes per cycle
- F02B2075/027—Engines characterised by their cycles, e.g. six-stroke having less than six strokes per cycle four
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/15—Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
- G01N2333/155—Lentiviridae, e.g. visna-maedi virus, equine infectious virus, FIV, SIV
- G01N2333/16—HIV-1, HIV-2
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/15—Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
- G01N2333/155—Lentiviridae, e.g. visna-maedi virus, equine infectious virus, FIV, SIV
- G01N2333/16—HIV-1, HIV-2
- G01N2333/163—Regulatory proteins, e.g. tat, nef, rev, vif, vpu, vpr, vpt, vpx
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/475—Assays involving growth factors
- G01N2333/50—Fibroblast growth factors [FGF]
- G01N2333/503—Fibroblast growth factors [FGF] basic FGF [bFGF]
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
- G01N2333/62—Insulins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/81—Protease inhibitors
- G01N2333/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- G01N2333/811—Serine protease (E.C. 3.4.21) inhibitors
- G01N2333/8121—Serpins
- G01N2333/8125—Alpha-1-antitrypsin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96433—Serine endopeptidases (3.4.21)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96433—Serine endopeptidases (3.4.21)
- G01N2333/96436—Granzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96433—Serine endopeptidases (3.4.21)
- G01N2333/96441—Serine endopeptidases (3.4.21) with definite EC number
- G01N2333/96455—Kallikrein (3.4.21.34; 3.4.21.35)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96486—Metalloendopeptidases (3.4.24)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/966—Elastase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/972—Plasminogen activators
- G01N2333/9726—Tissue plasminogen activator
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/974—Thrombin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/976—Trypsin; Chymotrypsin
Definitions
- This invention relates generally to a method for increasing the uptake of drugs into tumors by treatment with a nucleic acid ligand to PDGF in combination with a cytotoxic agent.
- the method used for identifying nucleic acid ligands to PDGF is called SELEX, an acronym for Systematic Evolution of Ligands by Exponential enrichment.
- the method of the present invention is useful for increasing the therapeutic effectiveness of cytotoxic agents.
- the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process is a method for the in vitro evolution of nucleic acid molecules with highly specific binding to target molecules and is described in U.S. patent application Ser. No. 07/536,428, filed Jun. 11, 1990, entitled “Systematic Evolution of Ligands by EXponential Enrichment,” now abandoned, U.S. Pat. No. 5,475,096, entitled “Nucleic Acid Ligands,” and U.S. Pat. No. 5,270,163 (see also WO 91/19813), entitled “Methods for Identifying Nucleic Acid Ligands,” each of which is specifically incorporated herein by reference in its entirety.
- Each of these applications, collectively referred to herein as the SELEX Patent Applications describes a fundamentally novel method for making a nucleic acid ligand to any desired target molecule.
- the SELEX process provides a class of products which are referred to as nucleic acid ligands or aptamers, each having a unique sequence, and which has the property of binding specifically to a desired target compound or molecule.
- Each SELEX-identified nucleic acid ligand is a specific ligand of a given target compound or molecule.
- the SELEX process is based on the unique insight that nucleic acids have sufficient capacity for forming a variety of two- and three-dimensional structures and sufficient chemical versatility available within their monomers to act as ligands (form specific binding pairs) with virtually any chemical compound, whether monomeric or polymeric. Molecules of any size or composition can serve as targets.
- the SELEX method applied to the application of high affinity binding involves selection from a mixture of candidate oligonucleotides and step-wise iterations of binding, partitioning and amplification, using the same general selection scheme, to achieve virtually any desired criterion of binding affinity and selectivity.
- the SELEX method includes steps of contacting the mixture with the target under conditions favorable for binding, partitioning unbound nucleic acids from those nucleic acids which have bound specifically to target molecules, dissociating the nucleic acid-target complexes, amplifying the nucleic acids dissociated from the nucleic acid-target complexes to yield a ligand enriched mixture of nucleic acids, then reiterating the steps of binding, partitioning, dissociating and amplifying through as many cycles as desired to yield highly specific high affinity nucleic acid ligands to the target molecule.
- nucleic acids as chemical compounds can form a wide array of shapes, sizes and configurations, and are capable of a far broader repertoire of binding and other functions than those displayed by nucleic acids in biological systems.
- the SELEX method encompasses the identification of high-affinity nucleic acid ligands containing modified nucleotides conferring improved characteristics on the ligand, such as improved in vivo stability or improved delivery characteristics. Examples of such modifications include chemical substitutions at the ribose and/or phosphate and/or base positions.
- SELEX process-identified nucleic acid ligands containing modified nucleotides are described in U.S. Pat. No. 5,660,985, entitled “High Affinity Nucleic Acid Ligands Containing Modified Nucleotides,” that describes oligonucleotides containing nucleotide derivatives chemically modified at the 5- and 2′-positions of pyrimidines.
- the SELEX method encompasses combining selected oligonucleotides with other selected oligonucleotides and non-oligonucleotide functional units as described in U.S. Pat. No. 5,637,459, entitled “Systematic Evolution of Ligands by EXponential Enrichment: Chimeric SELEX,” and U.S. Pat. No. 5,683,867, entitled “Systematic Evolution of Ligands by EXponential Enrichment: Blended SELEX,” respectively.
- These applications allow the combination of the broad array of shapes and other properties, and the efficient amplification and replication properties, of oligonucleotides with the desirable properties of other molecules.
- the SELEX method further encompasses combining selected nucleic acid ligands with lipophilic compounds or non-immunogenic, high molecular weight compounds in a diagnostic or therapeutic complex as described in U.S. Pat. No. 6,011,020, entitled “Nucleic Acid Ligand Complexes.”
- Platelet-derived growth factor (PDGF) and the cognate tyrosine kinase receptors are potent mitogens for mesenchymal cells.
- PDGF-BB is involved also in the regulation of IFP.
- IFP insulin-derived growth factor
- PDGF receptors are expressed in the tumor stroma of many common solid tumors, e.g. lung, colon and breast carcinomas. Based on these observations the effects of PDGF antagonists on tumor IFP, tumor transcapillary transport and therapeutic effects of cytotoxic drugs were investigated.
- the present invention includes a method for treating tumors, more specifically, solid tumors comprising administering to a host a therapeutically effective dose of a composition comprising a PDGF aptamer and a cytotoxic agent.
- the PDGF aptamer is identified using the SELEX process for the Systematic Evolution of Ligands by EXponential enrichment.
- the present invention also includes a method for reducing the interstitial fluid pressure (IFP) of a tumor, more specifically, a solid tumor comprising administering a PDGF nucleic acid ligand.
- IFP interstitial fluid pressure
- the present invention includes a method for increasing the uptake of cytotoxic agents into a tumor comprising administering to a host a composition comprising a PDGF aptamer and a cytotoxic agent.
- the present invention provides a novel method to increase drug uptake and therefore the therapeutic effectiveness of chemotherapy, by treatment with a PDGF inhibitor in conjunction with the therapeutic agent. As illustrated below, treatment with a PDGF aptamer decreases interstitial hypertension in these tumors thereby increasing the uptake of the therapeutic agent.
- FIGS. 1A and B show the molecular description of the PDGF aptamer used in the present study ((SEQ ID NO:1), FIG. 1A ) and the aptamer that was used as the control. ((SEQ ID NO:2), FIG. 1B ).
- FIG. 3 depicts the distribution of PDGF ⁇ -receptors and PDGF-AB/BB in PROb tumors.
- Morphological analysis of sections from PROb tumor cells revealed three discrete zones in the investigated tumors ( FIG. 3A ). Towards the exterior a cell rich zone containing proliferative cells arranged in glandular structures was evident. A zone further towards the central part of the tumors contained apoptotic cells and cell debris, whereas the central parts were largely acellular, but contained extracellular matrix deposits. Inflammatory infiltrates were not evident in the investigated tumors. Immunohistochemical analyses showed that PDGF-AB/BB was expressed in blood vessels ( FIG.
- FIG. 3B A weak but clearly visible staining of PDGF ⁇ -receptors was present in stromal structures in large parts of the tumors, especially around tumor glands ( FIG. 3D ). PDGF ⁇ -receptor staining was completely blocked by addition of a peptide corresponding to the amino acids 981-994 of the human/murine PDGF ⁇ -receptor at a 10-fold molar excess to the anti-PDGF ⁇ -receptor antibody ( FIG. 3E ).
- FIG. 4 illustrates graphically the reduction of IFP upon treatment of SCID-Mice bearing subcutaneous KAT-4 tumors (human anaplastic thyroid carcinoma with PDGF aptamers.
- FIG. 5 illustrates graphically the increased uptake of the chemotherapeutic agent Taxol upon pretreatment of KAT-4 tumors with PDGF aptamers.
- nucleic acid ligand is a non-naturally occurring nucleic acid having a desirable action on a target.
- Nucleic acid ligands are often referred to as “aptamers.”
- a desirable action includes, but is not limited to, binding of the target, catalytically changing the target, reacting with the target in a way which modifies/alters the target or the functional activity of the target, covalently attaching to the target as in a suicide inhibitor, facilitating the reaction between the target and another molecule.
- the action is specific binding affinity for a target molecule, such target molecule being a three dimensional chemical structure other than a polynucleotide that binds to the nucleic acid ligand through a mechanism which predominantly depends on Watson/Crick base pairing or triple helix binding, wherein the nucleic acid ligand does not have the known physiological function of being bound by the target molecule.
- the target is PDGF or regions thereof.
- Nucleic acid ligands include nucleic acids that are identified from a candidate mixture of nucleic acids, said nucleic acid ligand being a ligand of a given target, by the method comprising: a) contacting the candidate mixture with the target, wherein nucleic acids having an increased affinity to the target relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; b) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and c) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids.
- a “candidate mixture” is a mixture of nucleic acids of differing sequence from which to select a desired ligand.
- the source of a candidate mixture can be from naturally-occurring nucleic acids or fragments thereof, chemically synthesized nucleic acids, enzymatically synthesized nucleic acids or nucleic acids made by a combination of the foregoing techniques.
- each nucleic acid has fixed sequences surrounding a randomized region to facilitate the amplification process.
- nucleic acid means either DNA, RNA, single-stranded or double-stranded, and any chemical modifications thereof. Modifications include, but are not limited to, those which provide other chemical groups that incorporate additional charge, polarizability, hydrogen bonding, electrostatic interaction, and fluxionality to the nucleic acid ligand bases or to the nucleic acid ligand as a whole.
- modifications include, but are not limited to, 2′-position sugar modifications, 5-position pyrimidine modifications, 8-position purine modifications, modifications at exocyclic amines, substitution of 4-thiouridine, substitution of 5-bromo or 5-iodo-uracil; backbone modifications, methylations, unusual base-pairing combinations such as the isobases isocytidine and isoguanidine and the like. Modifications can also include 3′ and 5′ modifications such as capping.
- SELEX methodology involves the combination of selection of nucleic acid ligands that interact with a target in a desirable manner, for example binding to a protein, with amplification of those selected nucleic acids. Optional iterative cycling of the selection/amplification steps allows selection of one or a small number of nucleic acids which interact most strongly with the target from a pool which contains a very large number of nucleic acids. Cycling of the selection/amplification procedure is continued until a selected goal is achieved.
- the SELEX methodology was employed to obtain nucleic acid ligands to PDGF. See U.S. Pat. No. 5,674,685, issued Oct. 7, 1997, U.S. Pat. No. 5,668,264, issued Sep. 16, 1997 and U.S.
- SELEX target or “target” means any compound or molecule of interest for which a ligand is desired.
- a target can be a protein, peptide, carbohydrate, polysaccharide, glycoprotein, hormone, receptor, antigen, antibody, virus, substrate, metabolite, transition state analog, cofactor, inhibitor, drug, dye, nutrient, growth factor, etc. without limitation.
- the SELEX target was PDGF.
- a “cytotoxic agent” is any substance used to destroy tumor cells.
- the method of this invention can be used with any systemically administrated cytotoxic agent including, but not limited to, Bleomycin, Cisplatin, and Pt analogues; Carboplatin and Iproplatin, Cyclophosphamide, Daunorubicin, Doxofluoridine, Doxorubicin, Etoposide, Epirubicin, 5-Flurouracil, Gemzar, Ifosfamide, Melphalan, Methotrexate, Mithramycin, Mitomycin C, Mitoxanthrone, Streptozotocin, Taxol and Taxotere, Vincristine, Vinblastine, Vindesine, Vinorelbine, Topotecan and CPT-11.
- “Therapeutic” as used herein, includes treatment and/or prophylaxis. When used, therapeutic refers to humans, as well as, other animals.
- “Pharmaceutically or therapeutically effective dose or amount” refers to a dosage level sufficient to induce a desired biological result. That result may be the delivery of a pharmaceutical agent, alleviation of the signs, symptoms or causes of a disease or any other desirous alteration of a biological system.
- a “host” is a living subject, human or animal, into which a drug or cytotoxic agent is administered.
- the present invention includes a method for treating solid tumors comprising administering to a host a therapeutically effective dose of a composition comprising a PDGF aptamer and a cytotoxic agent.
- the PDGF aptamer is identified using the SELEX methodology.
- the SELEX process is described in U.S. patent application Ser. No. 07/536,428, entitled “Systematic Evolution of Ligands by Exponential Enrichment,” now abandoned, U.S. Pat. No. 5,475,096, entitled “Nucleic Acid Ligands,” and U.S. Pat. No. 5,270,163 (see also WO 91/19813), entitled “Methods for Identifying Nucleic Acid Ligands.”
- the SELEX process provides a class of products that are nucleic acid molecules, each having a unique sequence, and each of which has the property of binding specifically to a desired target compound or molecule.
- Target molecules are preferably proteins, but can also include among others carbohydrates, peptidoglycans and a variety of small molecules.
- SELEX methodology can also be used to target biological structures, such as cell surfaces or viruses, through specific interaction with a molecule that is an integral part of that biological structure.
- the SELEX process may be defined by the following series of steps.
- a candidate mixture of nucleic acids of differing sequence is prepared.
- the candidate mixture generally includes regions of fixed sequences (i.e., each of the members of the candidate mixture contains the same sequences in the same location) and regions of randomized sequences.
- the fixed sequence regions are selected either: (a) to assist in the amplification steps described below; (b) to mimic a sequence known to bind to the target; or (c) to enhance the concentration of a given structural arrangement of the nucleic acids in the candidate mixture.
- the randomized sequences can be totally randomized (i.e., the probability of finding a base at any position being one in four) or only partially randomized (e.g., the probability of finding a base at any location can be selected at any level between 0 and 100 percent).
- the candidate mixture is contacted with the selected target under conditions favorable for binding between the target and members of the candidate mixture. Under these circumstances, the interaction between the target and the nucleic acids of the candidate mixture can be considered as forming nucleic acid-target pairs between the target and those nucleic acids having the strongest affinity for the target.
- the nucleic acids with the highest affinity for the target are partitioned from those nucleic acids with lesser affinity to the target. Because only an extremely small number of sequences (and possibly only one molecule of nucleic acid) corresponding to the highest affinity nucleic acids exist in the candidate mixture, it is generally desirable to set the partitioning criteria so that a significant amount of the nucleic acids in the candidate mixture (approximately 5-50%) are retained during partitioning.
- nucleic acids selected during partitioning as having the relatively higher affinity for the target are then amplified to create a new candidate mixture that is enriched in nucleic acids having a relatively higher affinity for the target.
- the newly formed candidate mixture contains fewer and fewer unique sequences, and the average degree of affinity of the nucleic acids to the target will generally increase.
- the SELEX process will yield a candidate mixture containing one or a small number of unique nucleic acids representing those nucleic acids from the original candidate mixture having the highest affinity to the target molecule.
- the SELEX method encompasses the identification of high-affinity nucleic acid ligands containing modified nucleotides conferring improved characteristics on the ligand, such as improved in vivo stability or improved delivery characteristics. Examples of such modifications include chemical substitutions at the ribose and/or phosphate and/or base positions.
- SELEX-identified nucleic acid ligands containing modified nucleotides are described in U.S. Pat. No. 5,660,985, entitled “High Affinity Nucleic Acid Ligands Containing Modified Nucleotides,” that describes oligonucleotides containing nucleotide derivatives chemically modified at the 5- and 2′-positions of pyrimidines.
- the SELEX method encompasses combining selected oligonucleotides with other selected oligonucleotides and non-oligonucleotide functional units as described in U.S. Pat. No. 5,637,459, entitled “Systematic Evolution of Ligands by Exponential Enrichment: Chimeric SELEX,” and U.S. Pat. No. 5,683,867, entitled “Systematic Evolution of Ligands by Exponential Enrichment: Blended SELEX,” respectively.
- These applications allow the combination of the broad array of shapes and other properties, and the efficient amplification and replication properties, of oligonucleotides with the desirable properties of other molecules.
- nucleic acid ligand can be made to increase the in vivo stability of the nucleic acid ligand or to enhance or to mediate the delivery of the nucleic acid ligand. See, e.g., U.S. patent application Ser. No. 08/117,991, filed Sep. 8, 1993, now abandoned and U.S. Pat. No.
- nucleic acid ligands contemplated in this invention include, but are not limited to, those which provide other chemical groups that incorporate additional charge, polarizability, hydrophobicity, hydrogen bonding, electrostatic interaction, and fluxionality to the nucleic acid ligand bases or to the nucleic acid ligand as a whole.
- Such modifications include, but are not limited to, 2′-position sugar modifications, 5-position pyrimidine modifications, 8-position purine modifications, modifications at exocyclic amines, substitution of 4-thiouridine, substitution of 5-bromo or 5-iodo-uracil; backbone modifications, phosphorothioate or alkyl phosphate modifications, methylations, unusual base-pairing combinations such as the isobases, isocytidine and isoguanidine and the like. Modifications can also include 3′ and 5′ modifications such as capping.
- the nucleic acid ligands are RNA molecules that are 2′-fluoro (2′-F) modified on the sugar moiety of pyrimidine residues.
- the modifications can be pre- or post-SELEX process modifications.
- Pre-SELEX process modifications yield nucleic acid ligands with both specificity for their SELEX target and improved in vivo stability.
- Post-SELEX process modifications made to 2′-OH nucleic acid ligands can result in improved in vivo stability without adversely affecting the binding capacity of the nucleic acid ligand.
- nucleic acid ligands to PDGF of the invention are prepared through the SELEX methodology that is outlined above and thoroughly enabled in the SELEX applications incorporated herein by reference in their entirety.
- the cytotoxic agent can be any substance used in the prevention, diagnosis, alleviation, treatment or cure of disease. More specifically, the cytotoxic agent can be selected from any systemically administrated agent including, but not limited to, Bleomycin, Cisplatin, and Pt analogues; Carboplatin and Iproplatin, Cyclophosphamide, Daunorubicin, Doxofluoridine, Doxorubicin, Etoposide, Epirubicin, 5-Flurouracil, Gemzar, Ifosfamide, Melphalan, Methotrexate, Mithramycin, Mitomycin C, Mitoxanthrone, Streptozotocin, Taxol and Taxotere, Vincristine, Vinblastine, Vindesine, Vinorelbine, Topotecan and CPT-11.
- Bleomycin Cisplatin, and Pt analogues
- Carboplatin and Iproplatin Cyclophosphamide
- Daunorubicin Doxo
- Various delivery systems are known in the art and can be used to administer the therapeutic composition comprising the PDGF aptamer and cytotoxic agent of the invention, e.g., aqueous solution, encapsulation in liposomes, microparticles, and microcapsules.
- compositions of the invention may be administered parenterally by injection, although other effective administration forms, such as intraarticular injection, inhalant mists, orally active formulations, transdermal iontophoresis or suppositories are also envisioned.
- One preferred carrier is physiological saline solution, but it is contemplated that other pharmaceutically acceptable carriers may also be used.
- the carrier and the nucleic acid ligand constitute a physiologically-compatible, slow release formulation.
- the primary solvent in such a carrier may be either aqueous or non-aqueous in nature.
- the carrier may contain other pharmacologically-acceptable excipients for modifying or maintaining the pH, osmolarity, viscosity, clarity, color, sterility, stability, rate of dissolution, or odor of the formulation.
- the carrier may contain still other pharmacologically-acceptable excipients for modifying or maintaining the stability, rate of dissolution, release or absorption of the ligand.
- excipients are those substances usually and customarily employed to formulate dosages for parental administration in either unit dose or multi-dose form.
- the therapeutic composition may be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored either in a ready to use form or requiring reconstitution immediately prior to administration.
- the manner of administering formulations containing the compositions for systemic delivery may be via subcutaneous, intramuscular, intravenous, intranasal or vaginal or rectal suppository.
- the amount of the composition which will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder of condition, which can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness or advancement of the disease or condition, and should be decided according to the practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curved derived from in vitro or animal model test systems. For example, an effective amount of the composition of the invention is readily determined by administering graded doses of the composition of the invention and observing the desired effect.
- the PDGF-B aptamer used in the present study ((SEQ ID NO:1), FIG. 1A ) was produced by the SELEX method. (Tuerk and Gold (1990) Science 249:505-510, which is incorporated herein by reference in its entirety).
- the modified DNA aptamer, linked to 40 kDa polyethylene glycol, has a high affinity for PDGF-B with a Kd of ⁇ 0.1 nM. (Green et al. (1996) Biochemistry 35:14413-14424; Floege et al. (1999) Am. J. Pathol. 154:169-179; U.S. Pat. No. 6,229,002, issued May 8, 2001, ((SEQ ID NO:146), FIG.
- Example 1 describes the method used to treat PROb tumor-bearing rats with the PDGF-B specific aptamer.
- treatment of PROb tumor-bearing rats with the PDGF-B specific aptamer resulted in a decrease in tumor IFP when compared to rats treated with the control aptamer.
- the mean IFP in control aptamer-treated tumors was 14.6 ⁇ 1.2 mm Hg ( ⁇ S.E.M.) and 9.7 ⁇ 1.6 mm Hg ( ⁇ S.E.M.) in tumors treated with the PDGF-B specific aptamer.
- the method used to determine IFP is described in Example 2.
- PROb tumors were analyzed with regard to morphology, as well as distribution of PDGF-AB/BB and PDGF ⁇ -receptors as described in Example 3.
- the tumors displayed a heterogeneous morphology.
- tumor cells were arranged in glandular structures, whereas more centrally, tumor cells were less abundant and less well organized ( FIG. 3A ).
- the central part was basically acellular ( FIG. 3A ).
- Expression of PDGF-AB/BB in PROb tumors was found in blood vessels and possibly in extravascular stromal cells surrounding tumor glands ( FIG. 3B ). In the central part of the tumors, few if any tumor cells were present, but strongly PDGF-AB/BB positive cells were seen ( FIG. 3C ).
- PDGF-AB/BB positive tumor cells were found in vascular cells of larger vessels, and in unidentified, possibly microvascular, cells in the stroma ( FIG. 3D ).
- Tissue culture Cells were cultured under standard conditions and all tissue culture media were supplemented with 10% fetal bovine serum (FBS) and antibiotics, unless otherwise stated.
- FBS fetal bovine serum
- PAE cells were maintained in F12 culture medium (Sigma).
- PROb cells were kept in Dulbecco's Modified Eagle's Medium (Sigma).
- the PDGF-B aptamer used in the present study was produced by the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method. (Tuerk and Gold (1990) Science 249:505-510, which is incorporated herein by reference in its entirety).
- the modified DNA aptamer, linked to 40 kDa polyethylene glycol, has a high affinity for PDGF-B with a Kd of ⁇ 0.1 nM. (Green et al. (1996) Biochemistry 35:14413-14424; Floege et al. (1999) Am. J. Pathol. 154:169-179, each of which is incorporated herein by reference in its entirety).
- the aptamer shows biphasic clearance in rats following iv injection, approximately 47% is cleared with a half-life of 32 minutes, while the remainder is cleared with a half-life of 135 minutes. (Floege et al. (1999) Am. J. Pathol. 154:169-179). As a control, a sequence-scrambled analog of the PDGF-B aptamer was used. This oligonucleotide has a Kd for PDGF-BB in the micromolar range. (Floege et al. (1999) Am. J. Pathol. 154:169-179).
- Subcutaneously growing PROb tumors were established in BDIX rats by injection of 5 ⁇ 10 6 tumor cells in 50 ⁇ L of PBS in the flank.
- the rats were kept under pathogen-free conditions and were fed ad libitum. They were monitored regularly for tumor growth and experiments were performed 8-12 weeks after tumor cell implantation on rats bearing tumors ranging in size between 0.6 cm 3 and 7.6 cm 3 .
- the PDGF-B specific aptamer, and a control aptamer were given as i.p. injections in 2 mL PBS twice daily for 4 consecutive days at a dose of 7 mg ⁇ kg ⁇ 1 ⁇ day ⁇ 1 . All animal experiments described in the present report were approved by the Ethical Committee for Animal Experiments in Uppsala, Sweden.
- Tumor IFP was measured using the wick-in-needle technique (Wiig et al. (1982) Scan. J. Clin. Lab. Invest. 42:159-164). Briefly, rats were anaesthetized using isofluran in a mixture of O 2 and air. A standard 23-gauge needle filled with nylon-floss and saline, supplemented with 50 IE/mL of heparin was inserted into the center of the tumor and connected to a pressure transducer. This makeup enables continuous and stable recordings of fluid pressure. Fluid communication between the needle and the transducer was confirmed by compression and decompression of the tubing during each measurement.
- Tumor IFP was measured once before treatment with PDGF aptamers, and again 1-2 hours after the last administration of aptamers or vehicle alone. The change in tumor IFP was calculated for each tumor. After the second IFP measurement the rats were sacrificed and the tumors were excised and snap frozen in liquid nitrogen for further analyses.
- paraffin-embedded 4 ⁇ m sections were stained with van Gieson staining. Immunohistochemistry was performed on 6 ⁇ m cryosections from PROb tumors. Sections were fixed in acetone and blocked with 0.3% hydrogen peroxide in methanol for 15 minutes, rinsed and further incubated in a solution containing 20% human normal serum in a buffer containing 2% rat serum, 3% bovine serum albumin, 0.01% NP40 in PBS (RM buffer) for 5 hours at 4° C. Primary antibodies dissolved in RM buffer were added, either 4 ⁇ g/mL affinity-purified rabbit anti-PDGF preceptor IgG (Claesson-Welsh et al. (1988) Mol. Cell Biol.
- SCID-mice bearing subcutaneous KAT-4 tumors (human, anaplastic thyroid carcinoma) were pre-treated for 4 consecutive days with 12 mg ⁇ kg ⁇ 1 ⁇ day ⁇ 1 SELEX aptamers (i.p. injections, three times daily). Tumor interstitial fluid pressure was measured using the wick-in-needle technique. The results are set forth in FIG. 4 . * p ⁇ 0.05, Student's t-test.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Cell Biology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Gastroenterology & Hepatology (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Toxicology (AREA)
- Endocrinology (AREA)
- Plant Pathology (AREA)
- Bioinformatics & Computational Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nanotechnology (AREA)
Abstract
Description
- This application is a divisional of U.S. patent application Ser. No. 10/791,367, filed Mar. 2, 2004, which is a continuation of U.S. patent application Ser. No. 09/859,724, filed May 17, 2001, now U.S. Pat. No. 6,699,843, which claims the benefit of U.S. Provisional Application Ser. No. 60/205,006, filed May 17, 2000. U.S. patent application Ser. No. 09/859,724 is a continuation in part of U.S. patent application Ser. No. 08/479,725, filed Jun. 7, 1995, now U.S. Pat. No. 5,674,685, and also a continuation in part of U.S. patent application Ser. No. 08/479,783, filed Jun. 7, 1995, now U.S. Pat. No. 5,668,264, and also a continuation in part of U.S. patent application Ser. No. 08/618,693, filed Mar. 20, 1996, now U.S. Pat. No. 5,723,594, each entitled “High Affinity PDGF Nucleic Acid Ligands.” U.S. patent application Ser. No. 09/859,724 is also a continuation in part of U.S. patent application Ser. No. 08/991,743, filed Dec. 16, 1997, now U.S. Pat. No. 6,229,002, entitled “Platelet Derived Growth Factor (PDGF) Nucleic Acid Ligand Complexes.” Each of the foregoing applications is incorporated herein by reference in their entirety.
- This invention relates generally to a method for increasing the uptake of drugs into tumors by treatment with a nucleic acid ligand to PDGF in combination with a cytotoxic agent. The method used for identifying nucleic acid ligands to PDGF is called SELEX, an acronym for Systematic Evolution of Ligands by Exponential enrichment. The method of the present invention is useful for increasing the therapeutic effectiveness of cytotoxic agents.
- The Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process is a method for the in vitro evolution of nucleic acid molecules with highly specific binding to target molecules and is described in U.S. patent application Ser. No. 07/536,428, filed Jun. 11, 1990, entitled “Systematic Evolution of Ligands by EXponential Enrichment,” now abandoned, U.S. Pat. No. 5,475,096, entitled “Nucleic Acid Ligands,” and U.S. Pat. No. 5,270,163 (see also WO 91/19813), entitled “Methods for Identifying Nucleic Acid Ligands,” each of which is specifically incorporated herein by reference in its entirety. Each of these applications, collectively referred to herein as the SELEX Patent Applications, describes a fundamentally novel method for making a nucleic acid ligand to any desired target molecule.
- The SELEX process provides a class of products which are referred to as nucleic acid ligands or aptamers, each having a unique sequence, and which has the property of binding specifically to a desired target compound or molecule. Each SELEX-identified nucleic acid ligand is a specific ligand of a given target compound or molecule. The SELEX process is based on the unique insight that nucleic acids have sufficient capacity for forming a variety of two- and three-dimensional structures and sufficient chemical versatility available within their monomers to act as ligands (form specific binding pairs) with virtually any chemical compound, whether monomeric or polymeric. Molecules of any size or composition can serve as targets. The SELEX method applied to the application of high affinity binding involves selection from a mixture of candidate oligonucleotides and step-wise iterations of binding, partitioning and amplification, using the same general selection scheme, to achieve virtually any desired criterion of binding affinity and selectivity. Starting from a mixture of nucleic acids, preferably comprising a segment of randomized sequence, the SELEX method includes steps of contacting the mixture with the target under conditions favorable for binding, partitioning unbound nucleic acids from those nucleic acids which have bound specifically to target molecules, dissociating the nucleic acid-target complexes, amplifying the nucleic acids dissociated from the nucleic acid-target complexes to yield a ligand enriched mixture of nucleic acids, then reiterating the steps of binding, partitioning, dissociating and amplifying through as many cycles as desired to yield highly specific high affinity nucleic acid ligands to the target molecule.
- It has been recognized by the present inventors that the SELEX method demonstrates that nucleic acids as chemical compounds can form a wide array of shapes, sizes and configurations, and are capable of a far broader repertoire of binding and other functions than those displayed by nucleic acids in biological systems.
- The basic SELEX method has been modified to achieve a number of specific objectives. For example, U.S. patent application Ser. No. 07/960,093, filed Oct. 14, 1992, now abandoned, and U.S. Pat. No. 5,707,796, both entitled “Method for Selecting Nucleic Acids on the Basis of Structure,” describe the use of the SELEX process in conjunction with gel electrophoresis to select nucleic acid molecules with specific structural characteristics, such as bent DNA. U.S. patent application Ser. No. 08/123,935, filed Sep. 17, 1993, entitled “Photoselection of Nucleic Acid Ligands,” now abandoned, U.S. Pat. No. 5,763,177 and U.S. Pat. No. 6,011,577, both entitled “Systematic Evolution of Ligands by Exponential Enrichment: Photoselection of Nucleic Acid Ligands and Solution SELEX,” describe a SELEX based method for selecting nucleic acid ligands containing photoreactive groups capable of binding and/or photocrosslinking to and/or photoinactivating a target molecule. U.S. Pat. No. 5,580,737, entitled “High-Affinity Nucleic Acid Ligands That Discriminate Between Theophylline and Caffeine,” describes a method for identifying highly specific nucleic acid ligands able to discriminate between closely related molecules, which can be non-peptidic, termed Counter-SELEX. U.S. Pat. No. 5,567,588, entitled “Systematic Evolution of Ligands by EXponential Enrichment: Solution SELEX,” describes a SELEX-based method which achieves highly efficient partitioning between oligonucleotides having high and low affinity for a target molecule.
- The SELEX method encompasses the identification of high-affinity nucleic acid ligands containing modified nucleotides conferring improved characteristics on the ligand, such as improved in vivo stability or improved delivery characteristics. Examples of such modifications include chemical substitutions at the ribose and/or phosphate and/or base positions. SELEX process-identified nucleic acid ligands containing modified nucleotides are described in U.S. Pat. No. 5,660,985, entitled “High Affinity Nucleic Acid Ligands Containing Modified Nucleotides,” that describes oligonucleotides containing nucleotide derivatives chemically modified at the 5- and 2′-positions of pyrimidines. U.S. Pat. No. 5,580,737, supra, describes highly specific nucleic acid ligands containing one or more nucleotides modified with 2′-amino (2′-NH2), 2′-fluoro (2′-F), and/or 2′-O-methyl (2′-OMe). U.S. patent application Ser. No. 08/264,029, filed Jun. 22, 1994, entitled “Novel Method of Preparation of Known and
Novel 2′ Modified Nucleosides by Intramolecular Nucleophilic Displacement,” describes oligonucleotides containing various 2′-modified pyrimidines. - The SELEX method encompasses combining selected oligonucleotides with other selected oligonucleotides and non-oligonucleotide functional units as described in U.S. Pat. No. 5,637,459, entitled “Systematic Evolution of Ligands by EXponential Enrichment: Chimeric SELEX,” and U.S. Pat. No. 5,683,867, entitled “Systematic Evolution of Ligands by EXponential Enrichment: Blended SELEX,” respectively. These applications allow the combination of the broad array of shapes and other properties, and the efficient amplification and replication properties, of oligonucleotides with the desirable properties of other molecules.
- The SELEX method further encompasses combining selected nucleic acid ligands with lipophilic compounds or non-immunogenic, high molecular weight compounds in a diagnostic or therapeutic complex as described in U.S. Pat. No. 6,011,020, entitled “Nucleic Acid Ligand Complexes.” Each of the above described patent applications which describe modifications of the basic SELEX procedure are specifically incorporated by reference herein in their entirety.
- One approach to increasing the effectiveness of existing anti-cancer drugs for the treatment of solid malignancies is to augment the uptake of the drugs into tumors, and thereby obtain increased therapeutic concentration without elevating the adverse side-effects. Most solid tumors display an increased interstitial fluid pressure (IFP). The molecular mechanisms causing increased tumor IFP are poorly understood. However, tumor stroma involvement in IFP control has been demonstrated. (Gullino et al. (1964) Cancer Res. 24:780-797; Philips et al. (1990) J. Natl Cancer Inst. 82:1457-1469; Jain (1987) Cancer Res. 47:3039-3051). It has been suggested that high tumor IFP prevents drug transport from the circulation into the tumor interstitium. The reduction of tumor IFP therefore is a target for efforts to increase tumor drug uptake. (Jain (1996) Science 271:1079-1080). Several agents which induce a lowering of IFP, and thereby increase the transcapillary transport in experimental murine tumors have been identified, including nicotinamide (Lee et al. (1992) Cancer Res. 52:3237-3240), TNF-α (Kristensen et al. (1996) Br. J. Cancer 74:533-536) and dexamethasone (Kristjansen et al. (1993) Cancer Res. 53:4764-4766).
- Platelet-derived growth factor (PDGF) and the cognate tyrosine kinase receptors are potent mitogens for mesenchymal cells. In addition to its growth promoting effects, PDGF-BB is involved also in the regulation of IFP. After dextran-induced anaphylaxis and lowering of IFP in rat skin, local administration of PDGF-BB results in normalized IFP. PDGF receptors are expressed in the tumor stroma of many common solid tumors, e.g. lung, colon and breast carcinomas. Based on these observations the effects of PDGF antagonists on tumor IFP, tumor transcapillary transport and therapeutic effects of cytotoxic drugs were investigated.
- The present invention includes a method for treating tumors, more specifically, solid tumors comprising administering to a host a therapeutically effective dose of a composition comprising a PDGF aptamer and a cytotoxic agent. In a preferred embodiment the PDGF aptamer is identified using the SELEX process for the Systematic Evolution of Ligands by EXponential enrichment. The present invention also includes a method for reducing the interstitial fluid pressure (IFP) of a tumor, more specifically, a solid tumor comprising administering a PDGF nucleic acid ligand. Finally, the present invention includes a method for increasing the uptake of cytotoxic agents into a tumor comprising administering to a host a composition comprising a PDGF aptamer and a cytotoxic agent. The present invention provides a novel method to increase drug uptake and therefore the therapeutic effectiveness of chemotherapy, by treatment with a PDGF inhibitor in conjunction with the therapeutic agent. As illustrated below, treatment with a PDGF aptamer decreases interstitial hypertension in these tumors thereby increasing the uptake of the therapeutic agent.
-
FIGS. 1A and B show the molecular description of the PDGF aptamer used in the present study ((SEQ ID NO:1),FIG. 1A ) and the aptamer that was used as the control. ((SEQ ID NO:2),FIG. 1B ). -
FIG. 2 illustrates graphically the reduction in tumor interstitial fluid pressure (IFP) upon treatment of PROb tumors with a PDGF specific aptamer. Average tumor IFP in the animals treated with a sequence-scrambled control oligonucleotide (n=5) and PDGF-B specific aptamer (n=7) is depicted. Bars represent S.E.M; * p<0.05 -
FIG. 3 depicts the distribution of PDGF β-receptors and PDGF-AB/BB in PROb tumors. Morphological analysis of sections from PROb tumor cells revealed three discrete zones in the investigated tumors (FIG. 3A ). Towards the exterior a cell rich zone containing proliferative cells arranged in glandular structures was evident. A zone further towards the central part of the tumors contained apoptotic cells and cell debris, whereas the central parts were largely acellular, but contained extracellular matrix deposits. Inflammatory infiltrates were not evident in the investigated tumors. Immunohistochemical analyses showed that PDGF-AB/BB was expressed in blood vessels (FIG. 3B ) and in the central parts of tumors by cells morphologically identified as macrophages (FIG. 3C ). A weak but clearly visible staining of PDGF β-receptors was present in stromal structures in large parts of the tumors, especially around tumor glands (FIG. 3D ). PDGF β-receptor staining was completely blocked by addition of a peptide corresponding to the amino acids 981-994 of the human/murine PDGF β-receptor at a 10-fold molar excess to the anti-PDGF β-receptor antibody (FIG. 3E ). -
FIG. 4 illustrates graphically the reduction of IFP upon treatment of SCID-Mice bearing subcutaneous KAT-4 tumors (human anaplastic thyroid carcinoma with PDGF aptamers. -
FIG. 5 illustrates graphically the increased uptake of the chemotherapeutic agent Taxol upon pretreatment of KAT-4 tumors with PDGF aptamers. - Various terms are used herein to refer to aspects of the present invention. To aid in the clarification of the description of the components of this invention, the following definitions are provided.
- As used herein a “nucleic acid ligand” is a non-naturally occurring nucleic acid having a desirable action on a target. Nucleic acid ligands are often referred to as “aptamers.” A desirable action includes, but is not limited to, binding of the target, catalytically changing the target, reacting with the target in a way which modifies/alters the target or the functional activity of the target, covalently attaching to the target as in a suicide inhibitor, facilitating the reaction between the target and another molecule. In a preferred embodiment, the action is specific binding affinity for a target molecule, such target molecule being a three dimensional chemical structure other than a polynucleotide that binds to the nucleic acid ligand through a mechanism which predominantly depends on Watson/Crick base pairing or triple helix binding, wherein the nucleic acid ligand does not have the known physiological function of being bound by the target molecule. In the present invention, the target is PDGF or regions thereof. Nucleic acid ligands include nucleic acids that are identified from a candidate mixture of nucleic acids, said nucleic acid ligand being a ligand of a given target, by the method comprising: a) contacting the candidate mixture with the target, wherein nucleic acids having an increased affinity to the target relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; b) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and c) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids.
- As used herein a “candidate mixture” is a mixture of nucleic acids of differing sequence from which to select a desired ligand. The source of a candidate mixture can be from naturally-occurring nucleic acids or fragments thereof, chemically synthesized nucleic acids, enzymatically synthesized nucleic acids or nucleic acids made by a combination of the foregoing techniques. In a preferred embodiment, each nucleic acid has fixed sequences surrounding a randomized region to facilitate the amplification process.
- As used herein, “nucleic acid” means either DNA, RNA, single-stranded or double-stranded, and any chemical modifications thereof. Modifications include, but are not limited to, those which provide other chemical groups that incorporate additional charge, polarizability, hydrogen bonding, electrostatic interaction, and fluxionality to the nucleic acid ligand bases or to the nucleic acid ligand as a whole. Such modifications include, but are not limited to, 2′-position sugar modifications, 5-position pyrimidine modifications, 8-position purine modifications, modifications at exocyclic amines, substitution of 4-thiouridine, substitution of 5-bromo or 5-iodo-uracil; backbone modifications, methylations, unusual base-pairing combinations such as the isobases isocytidine and isoguanidine and the like. Modifications can also include 3′ and 5′ modifications such as capping.
- “SELEX” methodology involves the combination of selection of nucleic acid ligands that interact with a target in a desirable manner, for example binding to a protein, with amplification of those selected nucleic acids. Optional iterative cycling of the selection/amplification steps allows selection of one or a small number of nucleic acids which interact most strongly with the target from a pool which contains a very large number of nucleic acids. Cycling of the selection/amplification procedure is continued until a selected goal is achieved. In the present invention, the SELEX methodology was employed to obtain nucleic acid ligands to PDGF. See U.S. Pat. No. 5,674,685, issued Oct. 7, 1997, U.S. Pat. No. 5,668,264, issued Sep. 16, 1997 and U.S. Pat. No. 5,723,594, issued Mar. 3, 1998, each entitled “High Affinity PDGF Nucleic Acid Ligands,” and U.S. Pat. No. 6,229,002, issued, May 8, 2001, entitled “Platelet Derived Growth Factor (PDGF) Nucleic Acid Ligand Complexes.”
- The SELEX methodology is described in the SELEX Patent Applications.
- “SELEX target” or “target” means any compound or molecule of interest for which a ligand is desired. A target can be a protein, peptide, carbohydrate, polysaccharide, glycoprotein, hormone, receptor, antigen, antibody, virus, substrate, metabolite, transition state analog, cofactor, inhibitor, drug, dye, nutrient, growth factor, etc. without limitation. In this application, the SELEX target was PDGF.
- A “cytotoxic agent” is any substance used to destroy tumor cells. The method of this invention can be used with any systemically administrated cytotoxic agent including, but not limited to, Bleomycin, Cisplatin, and Pt analogues; Carboplatin and Iproplatin, Cyclophosphamide, Daunorubicin, Doxofluoridine, Doxorubicin, Etoposide, Epirubicin, 5-Flurouracil, Gemzar, Ifosfamide, Melphalan, Methotrexate, Mithramycin, Mitomycin C, Mitoxanthrone, Streptozotocin, Taxol and Taxotere, Vincristine, Vinblastine, Vindesine, Vinorelbine, Topotecan and CPT-11.
- “Therapeutic” as used herein, includes treatment and/or prophylaxis. When used, therapeutic refers to humans, as well as, other animals.
- “Pharmaceutically or therapeutically effective dose or amount” refers to a dosage level sufficient to induce a desired biological result. That result may be the delivery of a pharmaceutical agent, alleviation of the signs, symptoms or causes of a disease or any other desirous alteration of a biological system.
- A “host” is a living subject, human or animal, into which a drug or cytotoxic agent is administered.
- Note, that throughout this application various citations are provided. Each citation is specifically incorporated herein in its entirety by reference.
- The present invention includes a method for treating solid tumors comprising administering to a host a therapeutically effective dose of a composition comprising a PDGF aptamer and a cytotoxic agent. In a preferred embodiment the PDGF aptamer is identified using the SELEX methodology. The SELEX process is described in U.S. patent application Ser. No. 07/536,428, entitled “Systematic Evolution of Ligands by Exponential Enrichment,” now abandoned, U.S. Pat. No. 5,475,096, entitled “Nucleic Acid Ligands,” and U.S. Pat. No. 5,270,163 (see also WO 91/19813), entitled “Methods for Identifying Nucleic Acid Ligands.” These applications, each specifically incorporated herein by reference, are collectively called the SELEX Patent Applications.
- The SELEX process provides a class of products that are nucleic acid molecules, each having a unique sequence, and each of which has the property of binding specifically to a desired target compound or molecule. Target molecules are preferably proteins, but can also include among others carbohydrates, peptidoglycans and a variety of small molecules. SELEX methodology can also be used to target biological structures, such as cell surfaces or viruses, through specific interaction with a molecule that is an integral part of that biological structure.
- In its most basic form, the SELEX process may be defined by the following series of steps.
- 1. A candidate mixture of nucleic acids of differing sequence is prepared. The candidate mixture generally includes regions of fixed sequences (i.e., each of the members of the candidate mixture contains the same sequences in the same location) and regions of randomized sequences. The fixed sequence regions are selected either: (a) to assist in the amplification steps described below; (b) to mimic a sequence known to bind to the target; or (c) to enhance the concentration of a given structural arrangement of the nucleic acids in the candidate mixture. The randomized sequences can be totally randomized (i.e., the probability of finding a base at any position being one in four) or only partially randomized (e.g., the probability of finding a base at any location can be selected at any level between 0 and 100 percent).
- 2. The candidate mixture is contacted with the selected target under conditions favorable for binding between the target and members of the candidate mixture. Under these circumstances, the interaction between the target and the nucleic acids of the candidate mixture can be considered as forming nucleic acid-target pairs between the target and those nucleic acids having the strongest affinity for the target.
- 3. The nucleic acids with the highest affinity for the target are partitioned from those nucleic acids with lesser affinity to the target. Because only an extremely small number of sequences (and possibly only one molecule of nucleic acid) corresponding to the highest affinity nucleic acids exist in the candidate mixture, it is generally desirable to set the partitioning criteria so that a significant amount of the nucleic acids in the candidate mixture (approximately 5-50%) are retained during partitioning.
- 4. Those nucleic acids selected during partitioning as having the relatively higher affinity for the target are then amplified to create a new candidate mixture that is enriched in nucleic acids having a relatively higher affinity for the target.
- 5. By repeating the partitioning and amplifying steps above, the newly formed candidate mixture contains fewer and fewer unique sequences, and the average degree of affinity of the nucleic acids to the target will generally increase. Taken to its extreme, the SELEX process will yield a candidate mixture containing one or a small number of unique nucleic acids representing those nucleic acids from the original candidate mixture having the highest affinity to the target molecule.
- The basic SELEX method has been modified to achieve a number of specific objectives. For example, U.S. patent application Ser. No. 07/960,093, filed Oct. 14, 1992, now abandoned, and U.S. Pat. No. 5,707,796, both entitled “Method for Selecting Nucleic Acids on the Basis of Structure,” describe the use of the SELEX process in conjunction with gel electrophoresis to select nucleic acid molecules with specific structural characteristics, such as bent DNA. U.S. patent application Ser. No. 08/123,935, filed Sep. 17, 1993, entitled “Photoselection of Nucleic Acid Ligands,” now abandoned, U.S. Pat. No. 5,763,177 and U.S. Pat. No. 6,001,577, both entitled “Systematic Evolution of Ligands by Exponential Enrichment: Photoselection of Nucleic Acid Ligands and Solution SELEX,” all describe a SELEX based method for selecting nucleic acid ligands containing photoreactive groups capable of binding and/or photocrosslinking to and/or photoinactivating a target molecule. U.S. Pat. No. 5,580,737, entitled “High-Affinity Nucleic Acid Ligands That Discriminate Between Theophylline and Caffeine,” describes a method for identifying highly specific nucleic acid ligands able to discriminate between closely related molecules, termed Counter-SELEX. U.S. Pat. No. 5,567,588, entitled “Systematic Evolution of Ligands by Exponential Enrichment: Solution SELEX,” describes a SELEX-based method which achieves highly efficient partitioning between oligonucleotides having high and low affinity for a target molecule. U.S. Pat. No. 5,496,938, entitled “Nucleic Acid Ligands to HUV-RT and HIV-1 Rev,” describes methods for obtaining improved nucleic acid ligands after SELEX has been performed. U.S. Pat. No. 5,705,337, entitled “Systematic Evolution of Ligands by Exponential Enrichment: Chemi-SELEX,” describes methods for covalently linking a ligand to its target.
- The SELEX method encompasses the identification of high-affinity nucleic acid ligands containing modified nucleotides conferring improved characteristics on the ligand, such as improved in vivo stability or improved delivery characteristics. Examples of such modifications include chemical substitutions at the ribose and/or phosphate and/or base positions. SELEX-identified nucleic acid ligands containing modified nucleotides are described in U.S. Pat. No. 5,660,985, entitled “High Affinity Nucleic Acid Ligands Containing Modified Nucleotides,” that describes oligonucleotides containing nucleotide derivatives chemically modified at the 5- and 2′-positions of pyrimidines. U.S. Pat. No. 5,637,459, supra, describes highly specific nucleic acid ligands containing one or more nucleotides modified with 2′-amino (2′-NH2), 2′-fluoro (2′-F), and/or 2′-O-methyl (2′-OMe). U.S. patent application Ser. No. 08/264,029, filed Jun. 22, 1994, entitled “Novel Method of Preparation of Known and
Novel 2′ Modified Nucleosides by Intramolecular Nucleophilic Displacement,” describes oligonucleotides containing various 2′-modified pyrimidines. - The SELEX method encompasses combining selected oligonucleotides with other selected oligonucleotides and non-oligonucleotide functional units as described in U.S. Pat. No. 5,637,459, entitled “Systematic Evolution of Ligands by Exponential Enrichment: Chimeric SELEX,” and U.S. Pat. No. 5,683,867, entitled “Systematic Evolution of Ligands by Exponential Enrichment: Blended SELEX,” respectively. These applications allow the combination of the broad array of shapes and other properties, and the efficient amplification and replication properties, of oligonucleotides with the desirable properties of other molecules.
- In U.S. Pat. No. 5,496,938, methods are described for obtaining improved nucleic acid ligands after the SELEX process has been performed. This patent, entitled “Nucleic Acid Ligands to HIV-RT and HIV-1 Rev,” is specifically incorporated herein by reference.
- One potential problem encountered in the diagnostic use of nucleic acids is that oligonucleotides in their phosphodiester form may be quickly degraded in body fluids by intracellular and extracellular enzymes, such as endonucleases and exonucleases, before the desired effect is manifest. Certain chemical modifications of the nucleic acid ligand can be made to increase the in vivo stability of the nucleic acid ligand or to enhance or to mediate the delivery of the nucleic acid ligand. See, e.g., U.S. patent application Ser. No. 08/117,991, filed Sep. 8, 1993, now abandoned and U.S. Pat. No. 5,660,985, both entitled “High Affinity Nucleic Acid Ligands Containing Modified Nucleotides,” and U.S. patent application Ser. No. 09/362,578, filed Jul. 28, 1999, entitled “Transcription-free SELEX,” each of which is specifically incorporated herein by reference in its entirety. Modifications of the nucleic acid ligands contemplated in this invention include, but are not limited to, those which provide other chemical groups that incorporate additional charge, polarizability, hydrophobicity, hydrogen bonding, electrostatic interaction, and fluxionality to the nucleic acid ligand bases or to the nucleic acid ligand as a whole. Such modifications include, but are not limited to, 2′-position sugar modifications, 5-position pyrimidine modifications, 8-position purine modifications, modifications at exocyclic amines, substitution of 4-thiouridine, substitution of 5-bromo or 5-iodo-uracil; backbone modifications, phosphorothioate or alkyl phosphate modifications, methylations, unusual base-pairing combinations such as the isobases, isocytidine and isoguanidine and the like. Modifications can also include 3′ and 5′ modifications such as capping. In preferred embodiments of the instant invention, the nucleic acid ligands are RNA molecules that are 2′-fluoro (2′-F) modified on the sugar moiety of pyrimidine residues.
- The modifications can be pre- or post-SELEX process modifications. Pre-SELEX process modifications yield nucleic acid ligands with both specificity for their SELEX target and improved in vivo stability. Post-SELEX process modifications made to 2′-OH nucleic acid ligands can result in improved in vivo stability without adversely affecting the binding capacity of the nucleic acid ligand.
- Other modifications are known to one of ordinary skill in the art. Such modifications may be made post-SELEX process (modification of previously identified unmodified ligands) or by incorporation into the SELEX process.
- The nucleic acid ligands to PDGF of the invention are prepared through the SELEX methodology that is outlined above and thoroughly enabled in the SELEX applications incorporated herein by reference in their entirety.
- As noted above, the cytotoxic agent can be any substance used in the prevention, diagnosis, alleviation, treatment or cure of disease. More specifically, the cytotoxic agent can be selected from any systemically administrated agent including, but not limited to, Bleomycin, Cisplatin, and Pt analogues; Carboplatin and Iproplatin, Cyclophosphamide, Daunorubicin, Doxofluoridine, Doxorubicin, Etoposide, Epirubicin, 5-Flurouracil, Gemzar, Ifosfamide, Melphalan, Methotrexate, Mithramycin, Mitomycin C, Mitoxanthrone, Streptozotocin, Taxol and Taxotere, Vincristine, Vinblastine, Vindesine, Vinorelbine, Topotecan and CPT-11.
- Various delivery systems are known in the art and can be used to administer the therapeutic composition comprising the PDGF aptamer and cytotoxic agent of the invention, e.g., aqueous solution, encapsulation in liposomes, microparticles, and microcapsules.
- Therapeutic compositions of the invention may be administered parenterally by injection, although other effective administration forms, such as intraarticular injection, inhalant mists, orally active formulations, transdermal iontophoresis or suppositories are also envisioned. One preferred carrier is physiological saline solution, but it is contemplated that other pharmaceutically acceptable carriers may also be used. In one preferred embodiment, it is envisioned that the carrier and the nucleic acid ligand constitute a physiologically-compatible, slow release formulation. The primary solvent in such a carrier may be either aqueous or non-aqueous in nature. In addition, the carrier may contain other pharmacologically-acceptable excipients for modifying or maintaining the pH, osmolarity, viscosity, clarity, color, sterility, stability, rate of dissolution, or odor of the formulation. Similarly, the carrier may contain still other pharmacologically-acceptable excipients for modifying or maintaining the stability, rate of dissolution, release or absorption of the ligand. Such excipients are those substances usually and customarily employed to formulate dosages for parental administration in either unit dose or multi-dose form.
- Once the therapeutic composition has been formulated, it may be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored either in a ready to use form or requiring reconstitution immediately prior to administration. The manner of administering formulations containing the compositions for systemic delivery may be via subcutaneous, intramuscular, intravenous, intranasal or vaginal or rectal suppository.
- The amount of the composition which will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder of condition, which can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness or advancement of the disease or condition, and should be decided according to the practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curved derived from in vitro or animal model test systems. For example, an effective amount of the composition of the invention is readily determined by administering graded doses of the composition of the invention and observing the desired effect.
- The following examples are provided to explain and illustrate the present invention and are not intended to be limiting of the invention.
- The PDGF-B aptamer used in the present study ((SEQ ID NO:1),
FIG. 1A ) was produced by the SELEX method. (Tuerk and Gold (1990) Science 249:505-510, which is incorporated herein by reference in its entirety). The modified DNA aptamer, linked to 40 kDa polyethylene glycol, has a high affinity for PDGF-B with a Kd of ˜0.1 nM. (Green et al. (1996) Biochemistry 35:14413-14424; Floege et al. (1999) Am. J. Pathol. 154:169-179; U.S. Pat. No. 6,229,002, issued May 8, 2001, ((SEQ ID NO:146),FIG. 9A ), each of which is incorporated herein by reference in its entirety). A sequence-scrambled analog of the PDGF-B aptamer was used as a control. ((SEQ ID NO:2),FIG. 1B ). This oligonucleotide has a Kd for PDGF-BB in the micromolar range. (Floege et al. (1999) Am. J. Pathol. 154:169-179; U.S. Pat. No. 6,229,002, issued May 8, 2001, ((SEQ ID NO:147),FIG. 9B )). - Example 1 describes the method used to treat PROb tumor-bearing rats with the PDGF-B specific aptamer. As can be seen in
FIG. 2 treatment of PROb tumor-bearing rats with the PDGF-B specific aptamer resulted in a decrease in tumor IFP when compared to rats treated with the control aptamer. The mean IFP in control aptamer-treated tumors was 14.6±1.2 mm Hg (±S.E.M.) and 9.7±1.6 mm Hg (±S.E.M.) in tumors treated with the PDGF-B specific aptamer. The method used to determine IFP is described in Example 2. - PROb tumors were analyzed with regard to morphology, as well as distribution of PDGF-AB/BB and PDGF α-receptors as described in Example 3. The tumors displayed a heterogeneous morphology. At the tumor periphery, tumor cells were arranged in glandular structures, whereas more centrally, tumor cells were less abundant and less well organized (
FIG. 3A ). The central part was basically acellular (FIG. 3A ). Expression of PDGF-AB/BB in PROb tumors was found in blood vessels and possibly in extravascular stromal cells surrounding tumor glands (FIG. 3B ). In the central part of the tumors, few if any tumor cells were present, but strongly PDGF-AB/BB positive cells were seen (FIG. 3C ). In no part of the tumors could PDGF-AB/BB positive tumor cells be clearly discerned. PDGF β-receptors were found in vascular cells of larger vessels, and in unidentified, possibly microvascular, cells in the stroma (FIG. 3D ). The absence of PDGF-AB/BB and P-receptor expression by carcinoma cells in PROb tumors, is in agreement with the characteristics of cultured PROb cells (data not shown). - Prompted by these findings, the effects of treatment with PDGF aptamer (SEQ ID NO:1) were tested on the KAT-4 tumor model (Examples 4 and 5). Both of these tumor models showed PDGF receptor expression in tumor stroma but not on tumor cells. As can be seen in
FIGS. 4 and 5 treatment with PDGF aptamers lowers IFP in KAT-4 tumors and increases the uptake of Taxol. - Materials and Methods
- Tissue culture. Cells were cultured under standard conditions and all tissue culture media were supplemented with 10% fetal bovine serum (FBS) and antibiotics, unless otherwise stated. PAE cells were maintained in F12 culture medium (Sigma). PROb cells were kept in Dulbecco's Modified Eagle's Medium (Sigma).
- PDGF/PDGF receptor inhibitors. The PDGF-B aptamer used in the present study was produced by the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method. (Tuerk and Gold (1990) Science 249:505-510, which is incorporated herein by reference in its entirety). The modified DNA aptamer, linked to 40 kDa polyethylene glycol, has a high affinity for PDGF-B with a Kd of ˜0.1 nM. (Green et al. (1996) Biochemistry 35:14413-14424; Floege et al. (1999) Am. J. Pathol. 154:169-179, each of which is incorporated herein by reference in its entirety). The aptamer shows biphasic clearance in rats following iv injection, approximately 47% is cleared with a half-life of 32 minutes, while the remainder is cleared with a half-life of 135 minutes. (Floege et al. (1999) Am. J. Pathol. 154:169-179). As a control, a sequence-scrambled analog of the PDGF-B aptamer was used. This oligonucleotide has a Kd for PDGF-BB in the micromolar range. (Floege et al. (1999) Am. J. Pathol. 154:169-179).
- Subcutaneously growing PROb tumors (Martin et al. (1996) Int. J. Cancer 65:796-804) were established in BDIX rats by injection of 5×106 tumor cells in 50 μL of PBS in the flank. The rats were kept under pathogen-free conditions and were fed ad libitum. They were monitored regularly for tumor growth and experiments were performed 8-12 weeks after tumor cell implantation on rats bearing tumors ranging in size between 0.6 cm3 and 7.6 cm3. The PDGF-B specific aptamer, and a control aptamer (see above), were given as i.p. injections in 2 mL PBS twice daily for 4 consecutive days at a dose of 7 mg×kg−1×day−1. All animal experiments described in the present report were approved by the Ethical Committee for Animal Experiments in Uppsala, Sweden.
- Tumor IFP was measured using the wick-in-needle technique (Wiig et al. (1982) Scan. J. Clin. Lab. Invest. 42:159-164). Briefly, rats were anaesthetized using isofluran in a mixture of O2 and air. A standard 23-gauge needle filled with nylon-floss and saline, supplemented with 50 IE/mL of heparin was inserted into the center of the tumor and connected to a pressure transducer. This makeup enables continuous and stable recordings of fluid pressure. Fluid communication between the needle and the transducer was confirmed by compression and decompression of the tubing during each measurement. Tumor IFP was measured once before treatment with PDGF aptamers, and again 1-2 hours after the last administration of aptamers or vehicle alone. The change in tumor IFP was calculated for each tumor. After the second IFP measurement the rats were sacrificed and the tumors were excised and snap frozen in liquid nitrogen for further analyses.
- For routine morphology, paraffin-embedded 4 μm sections were stained with van Gieson staining. Immunohistochemistry was performed on 6 μm cryosections from PROb tumors. Sections were fixed in acetone and blocked with 0.3% hydrogen peroxide in methanol for 15 minutes, rinsed and further incubated in a solution containing 20% human normal serum in a buffer containing 2% rat serum, 3% bovine serum albumin, 0.01% NP40 in PBS (RM buffer) for 5 hours at 4° C. Primary antibodies dissolved in RM buffer were added, either 4 μg/mL affinity-purified rabbit anti-PDGF preceptor IgG (Claesson-Welsh et al. (1988) Mol. Cell Biol. 8:3476-3486) for 5 hours at 4° C., or overnight at 4° C. with 1.3 μg/mL of the monoclonal mouse anti-PDGF-AB/BB IgG (PGF 007, Mochida Pharmaceutical Company, Tokyo, Japan). Sections were rinsed in PBS with 0.01% NP40. Bound IgG was detected with biotinylated goat anti-rabbit or biotinylated rabbit anti-mouse antibodies, respectively. Sections were developed with a Vectastain ABC elite kit (Vector, Burlingame, Calif.) using amino-ethyl-carbazole as a chromophore. Sections were counter-stained with Mayer's hematoxylin for 30 seconds.
- SCID-mice bearing subcutaneous KAT-4 tumors (human, anaplastic thyroid carcinoma) were pre-treated for 4 consecutive days with 12 mg×kg−1×day−1 SELEX aptamers (i.p. injections, three times daily). Tumor interstitial fluid pressure was measured using the wick-in-needle technique. The results are set forth in
FIG. 4 . * p<0.05, Student's t-test. - SCID-mice bearing subcutaneous KAT-4 tumors (human, anaplastic thyroid carcinoma) were pre-treated for 4 consecutive days with 12 mg×kg−1×day−1 SELEX aptamers (i.p. injections, three times daily). [3H]Taxol was injected s.c at a site distant from the tumor and in a mix of 5 mg×kg−1 unlabelled Taxol. Eight or 24 hours following injection of radiolabelled drug, blood was sampled, animals were sacrificed and tumors and 4 other tissues were excised. Subsequently, tissues were weighed and homogenized in a RIPA lysis buffer and the amount radioactivity in each sample was determined in a scintillation counter. Tumor uptake of Taxol was expressed as cpm/g tumor tissue divided by cpm/ml blood. The results are set forth in
FIG. 5 . * p<0.05, Student's t-test. - Statistical analysis. Statistical analysis was performed using the paired or unpaired two-sided Student's t-test. A p-value <0.05 was considered statistically significant.
Claims (4)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11/518,321 US20070021327A1 (en) | 1995-06-07 | 2006-09-08 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
Applications Claiming Priority (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/479,783 US5668264A (en) | 1990-06-11 | 1995-06-07 | High affinity PDGF nucleic acid ligands |
US08/479,725 US5674685A (en) | 1990-06-11 | 1995-06-07 | High affinity PDGF nucleic acid ligands |
US08/618,693 US5723594A (en) | 1995-06-07 | 1996-03-20 | High affinity PDGF nucleic acid ligands |
US08/991,743 US6229002B1 (en) | 1995-06-07 | 1997-12-16 | Platelet derived growth factor (PDGF) nucleic acid ligand complexes |
US20500600P | 2000-05-17 | 2000-05-17 | |
US09/859,724 US6699843B2 (en) | 1995-06-07 | 2001-05-17 | Method for treatment of tumors using nucleic acid ligands to PDGF |
US10/791,367 US7141375B2 (en) | 1995-06-07 | 2004-03-02 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
US11/518,321 US20070021327A1 (en) | 1995-06-07 | 2006-09-08 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/791,367 Division US7141375B2 (en) | 1995-06-07 | 2004-03-02 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
Publications (1)
Publication Number | Publication Date |
---|---|
US20070021327A1 true US20070021327A1 (en) | 2007-01-25 |
Family
ID=33425824
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/859,724 Expired - Lifetime US6699843B2 (en) | 1995-06-07 | 2001-05-17 | Method for treatment of tumors using nucleic acid ligands to PDGF |
US10/791,367 Expired - Fee Related US7141375B2 (en) | 1995-06-07 | 2004-03-02 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
US11/518,321 Abandoned US20070021327A1 (en) | 1995-06-07 | 2006-09-08 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/859,724 Expired - Lifetime US6699843B2 (en) | 1995-06-07 | 2001-05-17 | Method for treatment of tumors using nucleic acid ligands to PDGF |
US10/791,367 Expired - Fee Related US7141375B2 (en) | 1995-06-07 | 2004-03-02 | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor |
Country Status (1)
Country | Link |
---|---|
US (3) | US6699843B2 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050096257A1 (en) * | 2003-08-27 | 2005-05-05 | David Shima | Combination therapy for the treatment of ocular neovascular disorders |
US8853376B2 (en) | 2002-11-21 | 2014-10-07 | Archemix Llc | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics |
US11273171B2 (en) | 2013-07-12 | 2022-03-15 | Iveric Bio, Inc. | Methods for treating or preventing ophthalmological conditions |
Families Citing this family (70)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6699843B2 (en) * | 1995-06-07 | 2004-03-02 | Gilead Sciences, Inc. | Method for treatment of tumors using nucleic acid ligands to PDGF |
JP2005533794A (en) * | 2002-06-18 | 2005-11-10 | アーケミックス コーポレイション | Aptamer-toxin molecules and methods of using the same |
CA2504633A1 (en) * | 2002-11-21 | 2004-06-10 | Archemix Corporation | Multivalent aptamer therapeutics with improved pharmacodynamic properties and methods of making and using the same |
US10100316B2 (en) * | 2002-11-21 | 2018-10-16 | Archemix Llc | Aptamers comprising CPG motifs |
US20050124565A1 (en) * | 2002-11-21 | 2005-06-09 | Diener John L. | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics |
US8039443B2 (en) | 2002-11-21 | 2011-10-18 | Archemix Corporation | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics |
GB0305422D0 (en) * | 2003-03-10 | 2003-04-16 | Univ Open | Detection, monitoring and treatment of cancer |
WO2005087808A2 (en) | 2004-03-05 | 2005-09-22 | Ludwig Institute For Cancer Research | Growth factor binding constructs materials and methods |
WO2005084412A2 (en) * | 2004-03-05 | 2005-09-15 | Archemix Corp. | Controlled modulation of the pharmacokinetics and biodistribution of aptamer therapeutics |
BRPI0508286B8 (en) | 2004-03-31 | 2021-05-25 | Dana Farber Cancer Inst Inc | method to determine the likelihood of efficacy of an egfr tyrosine kinase inhibitor to treat cancer, use of an egfr tyrosine kinase inhibitor, probe, kit, and, primer pair |
EP2436391A3 (en) * | 2004-11-02 | 2012-07-04 | Archemix LLC | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics |
DE502005009920D1 (en) * | 2004-11-29 | 2010-08-26 | Noxxon Pharma Ag | VASOPRESSIN BINDING L-NUCLEIC ACID |
CA2628451A1 (en) | 2005-11-04 | 2007-05-18 | Biogen Idec Ma Inc. | Methods for promoting neurite outgrowth and survival of dopaminergic neurons |
CA2913655A1 (en) | 2006-01-27 | 2007-08-09 | Biogen Ma Inc. | Nogo receptor antagonists |
US20080026947A1 (en) | 2006-02-08 | 2008-01-31 | Gmeiner William H | Cytotoxic nucleotides for targeted therapeutics |
JP2009529917A (en) | 2006-03-22 | 2009-08-27 | ヴァイラル ロジック システムズ テクノロジー コーポレーション | Methods for identifying target polypeptides and their use for the treatment of immunological disorders |
WO2007128109A1 (en) * | 2006-05-04 | 2007-11-15 | University Health Network | Aptamers that recognize the carbohydrate n-acetylgalactosamine (galnac) |
US8377448B2 (en) * | 2006-05-15 | 2013-02-19 | The Board Of Trustees Of The Leland Standford Junior University | CD47 related compositions and methods for treating immunological diseases and disorders |
EP2027151A2 (en) * | 2006-05-15 | 2009-02-25 | Viral Logic Systems Technology Corp. | Cd47 related compositions and methods for treating immunological diseases and disorders |
MX2008014953A (en) * | 2006-05-26 | 2009-03-05 | Bayer Healthcare Llc | Drug combinations with substituted diaryl ureas for the treatment of cancer. |
WO2008044068A2 (en) * | 2006-10-11 | 2008-04-17 | Fusion Antibodies Limited | Combination therapy |
US20100151465A1 (en) | 2008-03-27 | 2010-06-17 | Jingyue Ju | Selective Capture and Release of Analytes |
GB0807018D0 (en) * | 2008-04-17 | 2008-05-21 | Fusion Antibodies Ltd | Antibodies and treatment |
EP2304031A2 (en) * | 2008-06-11 | 2011-04-06 | Bionucleon S.r.l. | Inhibition of hrp-3 using modified oligonucleotides |
US9284559B2 (en) * | 2009-04-14 | 2016-03-15 | Wake Forest University Health Sciences | Multivalent aptamer complexes |
US9090663B2 (en) * | 2009-04-21 | 2015-07-28 | The Trustees Of Columbia University In The City Of New York | Systems and methods for the capture and separation of microparticles |
FR2948663B1 (en) * | 2009-07-31 | 2011-09-02 | Centre Nat Rech Scient | "APTAMERS THAT INHIBIT THE ACTIVITY OF THE HUMAN PROTEIN Rad51 AND THEIR BIOLOGICAL APPLICATIONS" |
WO2011058164A1 (en) | 2009-11-13 | 2011-05-19 | Pangaea Biotech, S.A. | Molecular biomarkers for predicting response to tyrosine kinase inhibitors in lung cancer |
WO2011106697A1 (en) | 2010-02-25 | 2011-09-01 | Schepens Eye Research Institute | Therapeutic compositions for the treatment of dry eye disease |
ES2575160T3 (en) | 2010-03-15 | 2016-06-24 | The Board Of Trustees Of The University Of Illinois | Inhibitors of the interactions that bind the alpha subunit of beta integrin-protein G |
JP2013545443A (en) * | 2010-10-14 | 2013-12-26 | リガド・バイオサイエンシーズ・インコーポレーテツド | CLEC-2 nucleic acid modulator |
WO2012087943A2 (en) | 2010-12-20 | 2012-06-28 | The Regents Of The University Of Michigan | Inhibitors of the epidermal growth factor receptor-heat shock protein 90 binding interaction |
EP2468883A1 (en) | 2010-12-22 | 2012-06-27 | Pangaea Biotech S.L. | Molecular biomarkers for predicting response to tyrosine kinase inhibitors in lung cancer |
EP2492688A1 (en) | 2011-02-23 | 2012-08-29 | Pangaea Biotech, S.A. | Molecular biomarkers for predicting response to antitumor treatment in lung cancer |
WO2013019714A1 (en) | 2011-07-29 | 2013-02-07 | The Trustees Of Columbia University In The City Of New York | Mems affinity sensor for continuous monitoring of analytes |
EP2764370B1 (en) | 2011-10-06 | 2017-09-13 | The Board of Trustees of the University of Illionis | Myosin binding protein-c for use in methods relating to diastolic heart failure |
WO2013055791A1 (en) | 2011-10-10 | 2013-04-18 | The Regents Of The University Of Michigan | Polymeric nanoparticles for ultrasound imaging and therapy |
WO2013190089A1 (en) | 2012-06-21 | 2013-12-27 | Pangaea Biotech, S.L. | Molecular biomarkers for predicting outcome in lung cancer |
WO2014012081A2 (en) | 2012-07-13 | 2014-01-16 | Ontorii, Inc. | Chiral control |
NO2733205T3 (en) | 2012-11-20 | 2018-06-09 | ||
SI2956476T1 (en) | 2013-02-18 | 2020-03-31 | Vegenics Pty Limited | Ligand binding molecules and uses thereof |
US9486533B2 (en) | 2013-09-27 | 2016-11-08 | Wake Forest University Health Sciences | Pharmaceutical compositions for high-capacity targeted delivery |
US10189908B2 (en) | 2014-02-05 | 2019-01-29 | The University Of Chicago | Chimeric antigen receptors recognizing cancer-specific TN glycopeptide variants |
AP2016009549A0 (en) | 2014-04-18 | 2016-11-30 | Acceleron Pharma Inc | Methods for increasing red blood cell levels and treating sickle-cell disease |
WO2015192111A1 (en) | 2014-06-13 | 2015-12-17 | Acceleron Pharma, Inc. | Methods and compositions for treating ulcers |
WO2016022696A1 (en) | 2014-08-05 | 2016-02-11 | The Trustees Of Columbia University In The City Of New York | Method of isolating aptamers for minimal residual disease detection |
MA41119A (en) | 2014-12-03 | 2017-10-10 | Acceleron Pharma Inc | METHODS OF TREATMENT OF MYELODYSPLASIC SYNDROMES AND SIDEROBLASTIC ANEMIA |
CN107683289B (en) | 2015-01-26 | 2021-08-06 | 芝加哥大学 | IL13R alpha 2 binding agents and their use in cancer treatment |
WO2016123143A1 (en) | 2015-01-26 | 2016-08-04 | The University Of Chicago | CAR T-CELLS RECOGNIZING CANCER-SPECIFIC IL 13Rα2 |
DK3050574T3 (en) | 2015-01-28 | 2020-01-20 | Univ Bordeaux | Use of plerixafor for the treatment and / or prevention of acute exacerbations of chronic obstructive pulmonary disease |
MA41919A (en) | 2015-04-06 | 2018-02-13 | Acceleron Pharma Inc | ALK4 HETEROMULTIMERS: ACTRIIB AND THEIR USES |
CN114736307A (en) | 2015-04-06 | 2022-07-12 | 阿塞勒隆制药公司 | TGF-beta superfamily type I and type II receptor heteromultimers and uses thereof |
US9884900B2 (en) | 2015-08-04 | 2018-02-06 | Acceleron Pharma Inc. | Methods for treating Janus kinase-associated disorders by administering soluble transforming growth factor beta type II receptor |
WO2017176265A1 (en) | 2016-04-06 | 2017-10-12 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
US10550170B2 (en) | 2015-11-23 | 2020-02-04 | Acceleron Pharma Inc. | Methods for treating vascular eye disorders with actrii antagonists |
RU2770006C2 (en) | 2016-05-16 | 2022-04-14 | Такеда Фармасьютикал Компани Лимитед | Ix padua factor antibodies |
ES2875905T3 (en) | 2016-07-15 | 2021-11-11 | Acceleron Pharma Inc | Compositions comprising ActRIIA polypeptides for use in the treatment of pulmonary hypertension |
CN109789184A (en) | 2016-07-27 | 2019-05-21 | 阿塞勒隆制药公司 | For treating the method and composition of myelofibrosis |
US11160876B2 (en) | 2016-09-01 | 2021-11-02 | Mayo Foundation For Medical Education And Research | Methods and compositions for targeting t-cell cancers |
CN109890419A (en) | 2016-09-01 | 2019-06-14 | 梅约医学教育与研究基金会 | Carrier-PD-L1 binding agent composition for treating cancer |
EP3522934A4 (en) | 2016-10-05 | 2020-04-15 | Acceleron Pharma Inc. | Compositions and method for treating kidney disease |
US11249082B2 (en) | 2016-10-29 | 2022-02-15 | University Of Miami | Zika virus assay systems |
CN106754936B (en) * | 2016-11-30 | 2019-06-07 | 吴冬 | The aptamer WYZ-2 and its screening technique of ovarian mucinous cancer cell 3AO and application |
CN106754935B (en) * | 2016-11-30 | 2019-04-30 | 吴冬 | The aptamer WYZ-5 and its screening technique of ovarian mucinous cancer cell 3AO and application |
WO2018164141A1 (en) | 2017-03-06 | 2018-09-13 | 学校法人東京女子医科大学 | Lypd1 inhibitor and method for producing biological tissue using same |
US11830582B2 (en) | 2018-06-14 | 2023-11-28 | University Of Miami | Methods of designing novel antibody mimetics for use in detecting antigens and as therapeutic agents |
JP7429294B2 (en) | 2020-07-13 | 2024-02-07 | 国立大学法人京都大学 | Skeletal muscle progenitor cells and methods for purifying the same, compositions for treating myogenic diseases, and methods for producing cell groups containing skeletal muscle progenitor cells |
TW202216778A (en) | 2020-07-15 | 2022-05-01 | 美商安進公司 | Tigit and cd112r blockade |
EP4208204A1 (en) | 2020-09-02 | 2023-07-12 | Mayo Foundation for Medical Education and Research | Antibody-nanoparticle complexes and methods for making and using the same |
WO2023137161A1 (en) | 2022-01-14 | 2023-07-20 | Amgen Inc. | Triple blockade of tigit, cd112r, and pd-l1 |
Citations (35)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4711955A (en) * | 1981-04-17 | 1987-12-08 | Yale University | Modified nucleotides and methods of preparing and using same |
US4828979A (en) * | 1984-11-08 | 1989-05-09 | Life Technologies, Inc. | Nucleotide analogs for nucleic acid labeling and detection |
US4863902A (en) * | 1985-11-28 | 1989-09-05 | Wakunaga Seiyaku Kabushiki Kaisha | Treatment of cancer |
US4904582A (en) * | 1987-06-11 | 1990-02-27 | Synthetic Genetics | Novel amphiphilic nucleic acid conjugates |
US4935363A (en) * | 1987-03-30 | 1990-06-19 | Board Of Regents, The University Of Texas System | Sterol regulatory elements |
US5070010A (en) * | 1989-10-30 | 1991-12-03 | Hoffman-La Roche Inc. | Method for determining anti-viral transactivating activity |
US5149794A (en) * | 1990-11-01 | 1992-09-22 | State Of Oregon | Covalent lipid-drug conjugates for drug targeting |
US5171217A (en) * | 1991-02-28 | 1992-12-15 | Indiana University Foundation | Method for delivery of smooth muscle cell inhibitors |
US5270163A (en) * | 1990-06-11 | 1993-12-14 | University Research Corporation | Methods for identifying nucleic acid ligands |
US5345022A (en) * | 1992-04-17 | 1994-09-06 | Chevron Research And Technology Company | Saturate minimization in normal alpha olefins |
US5425940A (en) * | 1986-04-09 | 1995-06-20 | Cetus Oncology Corporation | Combination therapy using interleukin-2 and tumor necrosis factor |
US5459015A (en) * | 1990-06-11 | 1995-10-17 | Nexstar Pharmaceuticals, Inc. | High-affinity RNA ligands of basic fibroblast growth factor |
US5475096A (en) * | 1990-06-11 | 1995-12-12 | University Research Corporation | Nucleic acid ligands |
US5476766A (en) * | 1990-06-11 | 1995-12-19 | Nexstar Pharmaceuticals, Inc. | Ligands of thrombin |
US5502037A (en) * | 1993-07-09 | 1996-03-26 | Neuromed Technologies, Inc. | Pro-cytotoxic drug conjugates for anticancer therapy |
US5521184A (en) * | 1992-04-03 | 1996-05-28 | Ciba-Geigy Corporation | Pyrimidine derivatives and processes for the preparation thereof |
US5563255A (en) * | 1994-05-31 | 1996-10-08 | Isis Pharmaceuticals, Inc. | Antisense oligonucleotide modulation of raf gene expression |
US5562922A (en) * | 1993-03-18 | 1996-10-08 | Cedars-Sinai Medical Center | Drug incorporating and release polymeric coating for bioprosthesis |
US5593974A (en) * | 1991-06-28 | 1997-01-14 | Massachusetts Institute Of Technology | Localized oligonucleotide therapy |
US5595877A (en) * | 1990-06-11 | 1997-01-21 | Nexstar Pharmaceuticals, Inc. | Methods of producing nucleic acid ligands |
US5614503A (en) * | 1993-11-12 | 1997-03-25 | Aronex Pharmaceuticals, Inc. | Amphipathic nucleic acid transporter |
US5631237A (en) * | 1992-12-22 | 1997-05-20 | Dzau; Victor J. | Method for producing in vivo delivery of therapeutic agents via liposomes |
US5668264A (en) * | 1990-06-11 | 1997-09-16 | Nexstar Pharmaceuticals, Inc. | High affinity PDGF nucleic acid ligands |
US5674685A (en) * | 1990-06-11 | 1997-10-07 | Nexstar Pharmaceuticals, Inc. | High affinity PDGF nucleic acid ligands |
US5723594A (en) * | 1995-06-07 | 1998-03-03 | Nexstar Pharmaceuticals, Inc. | High affinity PDGF nucleic acid ligands |
US5756291A (en) * | 1992-08-21 | 1998-05-26 | Gilead Sciences, Inc. | Aptamers specific for biomolecules and methods of making |
US5795910A (en) * | 1994-10-28 | 1998-08-18 | Cor Therapeutics, Inc. | Method and compositions for inhibiting protein kinases |
US5811533A (en) * | 1990-06-11 | 1998-09-22 | Nexstar Pharmaceuticals, Inc. | High-affinity oligonucleotide ligands to vascular endothelial growth factor (VEGF) |
US6011020A (en) * | 1990-06-11 | 2000-01-04 | Nexstar Pharmaceuticals, Inc. | Nucleic acid ligand complexes |
US6025507A (en) * | 1995-02-17 | 2000-02-15 | Schering Aktiengesellschaft | Borneol derivatives, methods of manufacturing them, and their pharmaceutical use |
US6054122A (en) * | 1990-11-27 | 2000-04-25 | The American National Red Cross | Supplemented and unsupplemented tissue sealants, methods of their production and use |
US6106866A (en) * | 1995-07-31 | 2000-08-22 | Access Pharmaceuticals, Inc. | In vivo agents comprising cationic drugs, peptides and metal chelators with acidic saccharides and glycosaminoglycans, giving improved site-selective localization, uptake mechanism, sensitivity and kinetic-spatial profiles, including tumor sites |
US6229002B1 (en) * | 1995-06-07 | 2001-05-08 | Nexstar Pharmaceuticlas, Inc. | Platelet derived growth factor (PDGF) nucleic acid ligand complexes |
US6699843B2 (en) * | 1995-06-07 | 2004-03-02 | Gilead Sciences, Inc. | Method for treatment of tumors using nucleic acid ligands to PDGF |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2183661B (en) | 1985-03-30 | 1989-06-28 | Marc Ballivet | Method for obtaining dna, rna, peptides, polypeptides or proteins by means of a dna recombinant technique |
WO1989006694A1 (en) | 1988-01-15 | 1989-07-27 | Trustees Of The University Of Pennsylvania | Process for selection of proteinaceous substances which mimic growth-inducing molecules |
JPH06503715A (en) | 1990-09-21 | 1994-04-28 | フレッド ハッチンソン キャンサー リサーチ センター | Protein sequence specific oligonucleotide sequences |
CA2104698A1 (en) | 1991-02-21 | 1992-08-22 | John J. Toole | Aptamers specific for biomolecules and methods of making |
-
2001
- 2001-05-17 US US09/859,724 patent/US6699843B2/en not_active Expired - Lifetime
-
2004
- 2004-03-02 US US10/791,367 patent/US7141375B2/en not_active Expired - Fee Related
-
2006
- 2006-09-08 US US11/518,321 patent/US20070021327A1/en not_active Abandoned
Patent Citations (37)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4711955A (en) * | 1981-04-17 | 1987-12-08 | Yale University | Modified nucleotides and methods of preparing and using same |
US4828979A (en) * | 1984-11-08 | 1989-05-09 | Life Technologies, Inc. | Nucleotide analogs for nucleic acid labeling and detection |
US4863902A (en) * | 1985-11-28 | 1989-09-05 | Wakunaga Seiyaku Kabushiki Kaisha | Treatment of cancer |
US4683195B1 (en) * | 1986-01-30 | 1990-11-27 | Cetus Corp | |
US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US5425940A (en) * | 1986-04-09 | 1995-06-20 | Cetus Oncology Corporation | Combination therapy using interleukin-2 and tumor necrosis factor |
US4935363A (en) * | 1987-03-30 | 1990-06-19 | Board Of Regents, The University Of Texas System | Sterol regulatory elements |
US4904582A (en) * | 1987-06-11 | 1990-02-27 | Synthetic Genetics | Novel amphiphilic nucleic acid conjugates |
US5070010A (en) * | 1989-10-30 | 1991-12-03 | Hoffman-La Roche Inc. | Method for determining anti-viral transactivating activity |
US5475096A (en) * | 1990-06-11 | 1995-12-12 | University Research Corporation | Nucleic acid ligands |
US5811533A (en) * | 1990-06-11 | 1998-09-22 | Nexstar Pharmaceuticals, Inc. | High-affinity oligonucleotide ligands to vascular endothelial growth factor (VEGF) |
US5674685A (en) * | 1990-06-11 | 1997-10-07 | Nexstar Pharmaceuticals, Inc. | High affinity PDGF nucleic acid ligands |
US5668264A (en) * | 1990-06-11 | 1997-09-16 | Nexstar Pharmaceuticals, Inc. | High affinity PDGF nucleic acid ligands |
US5459015A (en) * | 1990-06-11 | 1995-10-17 | Nexstar Pharmaceuticals, Inc. | High-affinity RNA ligands of basic fibroblast growth factor |
US5595877A (en) * | 1990-06-11 | 1997-01-21 | Nexstar Pharmaceuticals, Inc. | Methods of producing nucleic acid ligands |
US5476766A (en) * | 1990-06-11 | 1995-12-19 | Nexstar Pharmaceuticals, Inc. | Ligands of thrombin |
US5270163A (en) * | 1990-06-11 | 1993-12-14 | University Research Corporation | Methods for identifying nucleic acid ligands |
US6011020A (en) * | 1990-06-11 | 2000-01-04 | Nexstar Pharmaceuticals, Inc. | Nucleic acid ligand complexes |
US5149794A (en) * | 1990-11-01 | 1992-09-22 | State Of Oregon | Covalent lipid-drug conjugates for drug targeting |
US6054122A (en) * | 1990-11-27 | 2000-04-25 | The American National Red Cross | Supplemented and unsupplemented tissue sealants, methods of their production and use |
US5171217A (en) * | 1991-02-28 | 1992-12-15 | Indiana University Foundation | Method for delivery of smooth muscle cell inhibitors |
US5593974A (en) * | 1991-06-28 | 1997-01-14 | Massachusetts Institute Of Technology | Localized oligonucleotide therapy |
US5521184A (en) * | 1992-04-03 | 1996-05-28 | Ciba-Geigy Corporation | Pyrimidine derivatives and processes for the preparation thereof |
US5345022A (en) * | 1992-04-17 | 1994-09-06 | Chevron Research And Technology Company | Saturate minimization in normal alpha olefins |
US5756291A (en) * | 1992-08-21 | 1998-05-26 | Gilead Sciences, Inc. | Aptamers specific for biomolecules and methods of making |
US5631237A (en) * | 1992-12-22 | 1997-05-20 | Dzau; Victor J. | Method for producing in vivo delivery of therapeutic agents via liposomes |
US5562922A (en) * | 1993-03-18 | 1996-10-08 | Cedars-Sinai Medical Center | Drug incorporating and release polymeric coating for bioprosthesis |
US5502037A (en) * | 1993-07-09 | 1996-03-26 | Neuromed Technologies, Inc. | Pro-cytotoxic drug conjugates for anticancer therapy |
US5614503A (en) * | 1993-11-12 | 1997-03-25 | Aronex Pharmaceuticals, Inc. | Amphipathic nucleic acid transporter |
US5563255A (en) * | 1994-05-31 | 1996-10-08 | Isis Pharmaceuticals, Inc. | Antisense oligonucleotide modulation of raf gene expression |
US5795910A (en) * | 1994-10-28 | 1998-08-18 | Cor Therapeutics, Inc. | Method and compositions for inhibiting protein kinases |
US6025507A (en) * | 1995-02-17 | 2000-02-15 | Schering Aktiengesellschaft | Borneol derivatives, methods of manufacturing them, and their pharmaceutical use |
US5723594A (en) * | 1995-06-07 | 1998-03-03 | Nexstar Pharmaceuticals, Inc. | High affinity PDGF nucleic acid ligands |
US6229002B1 (en) * | 1995-06-07 | 2001-05-08 | Nexstar Pharmaceuticlas, Inc. | Platelet derived growth factor (PDGF) nucleic acid ligand complexes |
US6582918B2 (en) * | 1995-06-07 | 2003-06-24 | Gilead Sciences, Inc. | Platelet derived growth factor (PDGF) nucleic acid ligand complexes |
US6699843B2 (en) * | 1995-06-07 | 2004-03-02 | Gilead Sciences, Inc. | Method for treatment of tumors using nucleic acid ligands to PDGF |
US6106866A (en) * | 1995-07-31 | 2000-08-22 | Access Pharmaceuticals, Inc. | In vivo agents comprising cationic drugs, peptides and metal chelators with acidic saccharides and glycosaminoglycans, giving improved site-selective localization, uptake mechanism, sensitivity and kinetic-spatial profiles, including tumor sites |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8853376B2 (en) | 2002-11-21 | 2014-10-07 | Archemix Llc | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics |
US20050096257A1 (en) * | 2003-08-27 | 2005-05-05 | David Shima | Combination therapy for the treatment of ocular neovascular disorders |
US20100119522A1 (en) * | 2003-08-27 | 2010-05-13 | Ophthotech Corporation | Combination therapy for the treatment of ocular neovascular disorders |
US20100129364A1 (en) * | 2003-08-27 | 2010-05-27 | Ophthotech Corporation | Combination therapy for the treatment of ocular neovascular disorders |
US7759472B2 (en) | 2003-08-27 | 2010-07-20 | Ophthotech Corporation | Combination therapy for the treatment of ocular neovascular disorders |
US20110200593A1 (en) * | 2003-08-27 | 2011-08-18 | Ophthotech Corporation | Combination Therapy for the Treatment of Ocular Neovascular Disorders |
US8187597B2 (en) | 2003-08-27 | 2012-05-29 | Ophthotech Corporation | Combination therapy for the treatment of ocular neovascular disorders |
US8206707B2 (en) | 2003-08-27 | 2012-06-26 | Ophthotech Corporation | Combination therapy for the treatment of ocular neovascular disorders |
US8685397B2 (en) | 2003-08-27 | 2014-04-01 | Ophthotech Corporation | Combination therapy for the treatment of ocular neovascular disorders |
US11273171B2 (en) | 2013-07-12 | 2022-03-15 | Iveric Bio, Inc. | Methods for treating or preventing ophthalmological conditions |
Also Published As
Publication number | Publication date |
---|---|
US20020034506A1 (en) | 2002-03-21 |
US6699843B2 (en) | 2004-03-02 |
US7141375B2 (en) | 2006-11-28 |
US20040224335A1 (en) | 2004-11-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US7141375B2 (en) | Methods and compositions for treatment of tumors using nucleic acid ligands to platelet-derived growth factor | |
AU777043B2 (en) | Nucleic acid ligands to CD40ligand | |
US6124449A (en) | High affinity TGFβ nucleic acid ligands and inhibitors | |
ES2426160T3 (en) | Platelet-derived growth factor nucleic acid ligand complexes (PDGF) | |
JP4176466B2 (en) | Nucleic acid ligands for prostate specific membrane antigen | |
AU767501B2 (en) | Tenascin-C nucleic acid ligands | |
US20040258656A1 (en) | High affinity TGFbeta nucleic acid ligands and inhibitors | |
US20040009510A1 (en) | Allosteric nucleic acid sensor molecules | |
WO1999007724A1 (en) | Systematic evolution of ligands by exponential enrichment: tissue selex | |
RU2571660C2 (en) | Modifiers of glycoprotein vi representing nucleic acid | |
US20040253679A1 (en) | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics | |
WO2001087351A1 (en) | Method for treatment of tumors using nucleic acid ligands to pdgf | |
US8853376B2 (en) | Stabilized aptamers to platelet derived growth factor and their use as oncology therapeutics | |
US7005260B1 (en) | Tenascin-C nucleic acid ligands | |
US20030065155A1 (en) | Nucleic acid sensor molecules | |
US20030008295A1 (en) | Nucleic acid sensor molecules |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: GILEAD SCIENCES, INC., CALIFORNIA Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:LUDWIG INSTITUTE FOR CANCER RESEARCH;REEL/FRAME:018332/0337 Effective date: 20011212 Owner name: GILEAD SCIENCES, INC., CALIFORNIA Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:RUBIN, KRISTOFER;REEL/FRAME:018332/0349 Effective date: 20011211 Owner name: LUDWIG INSTITUTE FOR CANCER RESEARCH, UNITED KINGD Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:PIETRAS, KRISTIAN;OSTMAN, ARNE;HELDIN, CARL-HENRIK;REEL/FRAME:018332/0174 Effective date: 20011211 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |