WO2009155556A2 - Crkl targeting peptides - Google Patents
Crkl targeting peptides Download PDFInfo
- Publication number
- WO2009155556A2 WO2009155556A2 PCT/US2009/048024 US2009048024W WO2009155556A2 WO 2009155556 A2 WO2009155556 A2 WO 2009155556A2 US 2009048024 W US2009048024 W US 2009048024W WO 2009155556 A2 WO2009155556 A2 WO 2009155556A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- peptide
- crkl
- protein
- seq
- tumor
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 299
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 80
- 230000008685 targeting Effects 0.000 title claims abstract description 80
- 101150118364 Crkl gene Proteins 0.000 title 1
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 189
- 238000000034 method Methods 0.000 claims abstract description 88
- 201000011510 cancer Diseases 0.000 claims abstract description 44
- 239000000203 mixture Substances 0.000 claims abstract description 40
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 27
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 27
- 102100029375 Crk-like protein Human genes 0.000 claims abstract 13
- 101000919315 Homo sapiens Crk-like protein Proteins 0.000 claims abstract 13
- 210000004027 cell Anatomy 0.000 claims description 163
- 108090000623 proteins and genes Proteins 0.000 claims description 143
- 102000004169 proteins and genes Human genes 0.000 claims description 124
- 230000027455 binding Effects 0.000 claims description 78
- 102000006495 integrins Human genes 0.000 claims description 78
- 108010044426 integrins Proteins 0.000 claims description 78
- 239000003795 chemical substances by application Substances 0.000 claims description 55
- 150000001413 amino acids Chemical class 0.000 claims description 28
- 238000002864 sequence alignment Methods 0.000 claims description 27
- 210000001519 tissue Anatomy 0.000 claims description 25
- 239000002502 liposome Substances 0.000 claims description 21
- 230000006907 apoptotic process Effects 0.000 claims description 20
- 239000003814 drug Substances 0.000 claims description 20
- 241000700605 Viruses Species 0.000 claims description 15
- 102400001047 Endostatin Human genes 0.000 claims description 12
- 108010079505 Endostatins Proteins 0.000 claims description 12
- 229940127089 cytotoxic agent Drugs 0.000 claims description 12
- 241000701161 unidentified adenovirus Species 0.000 claims description 11
- 239000004037 angiogenesis inhibitor Substances 0.000 claims description 10
- 229940079593 drug Drugs 0.000 claims description 10
- 102000004127 Cytokines Human genes 0.000 claims description 9
- 108090000695 Cytokines Proteins 0.000 claims description 9
- 229930012538 Paclitaxel Natural products 0.000 claims description 9
- 239000003242 anti bacterial agent Substances 0.000 claims description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 9
- 239000012634 fragment Substances 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
- 238000002560 therapeutic procedure Methods 0.000 claims description 9
- 102000000589 Interleukin-1 Human genes 0.000 claims description 8
- 108010002352 Interleukin-1 Proteins 0.000 claims description 8
- 206010060862 Prostate cancer Diseases 0.000 claims description 8
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 8
- 239000002246 antineoplastic agent Substances 0.000 claims description 8
- 239000005556 hormone Substances 0.000 claims description 8
- 229940088597 hormone Drugs 0.000 claims description 8
- 239000012216 imaging agent Substances 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 claims description 7
- 108010069514 Cyclic Peptides Proteins 0.000 claims description 7
- 102000001189 Cyclic Peptides Human genes 0.000 claims description 7
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 claims description 7
- 239000000427 antigen Substances 0.000 claims description 7
- 108091007433 antigens Proteins 0.000 claims description 7
- 102000036639 antigens Human genes 0.000 claims description 7
- 230000004083 survival effect Effects 0.000 claims description 7
- 229960004528 vincristine Drugs 0.000 claims description 7
- 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 7
- 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 7
- 102400000068 Angiostatin Human genes 0.000 claims description 6
- 108010079709 Angiostatins Proteins 0.000 claims description 6
- 108010006654 Bleomycin Proteins 0.000 claims description 6
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims description 6
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 claims description 6
- 101000605403 Homo sapiens Plasminogen Proteins 0.000 claims description 6
- 108010065805 Interleukin-12 Proteins 0.000 claims description 6
- 102000013462 Interleukin-12 Human genes 0.000 claims description 6
- 102000004211 Platelet factor 4 Human genes 0.000 claims description 6
- 108090000778 Platelet factor 4 Proteins 0.000 claims description 6
- 108060008245 Thrombospondin Proteins 0.000 claims description 6
- 102000002938 Thrombospondin Human genes 0.000 claims description 6
- 102000016549 Vascular Endothelial Growth Factor Receptor-2 Human genes 0.000 claims description 6
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 claims description 6
- FZCSTZYAHCUGEM-UHFFFAOYSA-N aspergillomarasmine B Natural products OC(=O)CNC(C(O)=O)CNC(C(O)=O)CC(O)=O FZCSTZYAHCUGEM-UHFFFAOYSA-N 0.000 claims description 6
- 229960001561 bleomycin Drugs 0.000 claims description 6
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound 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 6
- 239000002254 cytotoxic agent Substances 0.000 claims description 6
- 231100000599 cytotoxic agent Toxicity 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 6
- 239000003102 growth factor Substances 0.000 claims description 6
- 230000011664 signaling Effects 0.000 claims description 6
- 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 5
- 210000004556 brain Anatomy 0.000 claims description 5
- 239000000032 diagnostic agent Substances 0.000 claims description 5
- 229940039227 diagnostic agent Drugs 0.000 claims description 5
- 229960003668 docetaxel Drugs 0.000 claims description 5
- 241001430294 unidentified retrovirus Species 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 4
- 108010067306 Fibronectins Proteins 0.000 claims description 4
- 102000016359 Fibronectins Human genes 0.000 claims description 4
- 102000008070 Interferon-gamma Human genes 0.000 claims description 4
- 108010074328 Interferon-gamma Proteins 0.000 claims description 4
- 108010002350 Interleukin-2 Proteins 0.000 claims description 4
- 206010027406 Mesothelioma Diseases 0.000 claims description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 4
- 241000700618 Vaccinia virus Species 0.000 claims description 4
- 239000011324 bead Substances 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 4
- 210000004962 mammalian cell Anatomy 0.000 claims description 4
- 230000004962 physiological condition Effects 0.000 claims description 4
- 102000003390 tumor necrosis factor Human genes 0.000 claims description 4
- 241001529453 unidentified herpesvirus Species 0.000 claims description 4
- 241001515965 unidentified phage Species 0.000 claims description 4
- UEJJHQNACJXSKW-UHFFFAOYSA-N 2-(2,6-dioxopiperidin-3-yl)-1H-isoindole-1,3(2H)-dione Chemical compound O=C1C2=CC=CC=C2C(=O)N1C1CCC(=O)NC1=O UEJJHQNACJXSKW-UHFFFAOYSA-N 0.000 claims description 3
- CQOQDQWUFQDJMK-SSTWWWIQSA-N 2-methoxy-17beta-estradiol Chemical compound C([C@@H]12)C[C@]3(C)[C@@H](O)CC[C@H]3[C@@H]1CCC1=C2C=C(OC)C(O)=C1 CQOQDQWUFQDJMK-SSTWWWIQSA-N 0.000 claims description 3
- SGOOQMRIPALTEL-UHFFFAOYSA-N 4-hydroxy-N,1-dimethyl-2-oxo-N-phenyl-3-quinolinecarboxamide Chemical compound OC=1C2=CC=CC=C2N(C)C(=O)C=1C(=O)N(C)C1=CC=CC=C1 SGOOQMRIPALTEL-UHFFFAOYSA-N 0.000 claims description 3
- 108010048036 Angiopoietin-2 Proteins 0.000 claims description 3
- 108010064733 Angiotensins Proteins 0.000 claims description 3
- 102000015427 Angiotensins Human genes 0.000 claims description 3
- 208000026310 Breast neoplasm Diseases 0.000 claims description 3
- 102100039398 C-X-C motif chemokine 2 Human genes 0.000 claims description 3
- 102100036170 C-X-C motif chemokine 9 Human genes 0.000 claims description 3
- 101710085500 C-X-C motif chemokine 9 Proteins 0.000 claims description 3
- 108050004290 Cecropin Proteins 0.000 claims description 3
- VWFCHDSQECPREK-LURJTMIESA-N Cidofovir Chemical compound NC=1C=CN(C[C@@H](CO)OCP(O)(O)=O)C(=O)N=1 VWFCHDSQECPREK-LURJTMIESA-N 0.000 claims description 3
- 150000008574 D-amino acids Chemical class 0.000 claims description 3
- 108010002069 Defensins Proteins 0.000 claims description 3
- 102000000541 Defensins Human genes 0.000 claims description 3
- 101000827763 Drosophila melanogaster Fibroblast growth factor receptor homolog 1 Proteins 0.000 claims description 3
- 108010026389 Gramicidin Proteins 0.000 claims description 3
- 102000013271 Hemopexin Human genes 0.000 claims description 3
- 108010026027 Hemopexin Proteins 0.000 claims description 3
- MCAHMSDENAOJFZ-UHFFFAOYSA-N Herbimycin A Natural products N1C(=O)C(C)=CC=CC(OC)C(OC(N)=O)C(C)=CC(C)C(OC)C(OC)CC(C)C(OC)C2=CC(=O)C=C1C2=O MCAHMSDENAOJFZ-UHFFFAOYSA-N 0.000 claims description 3
- 101000889128 Homo sapiens C-X-C motif chemokine 2 Proteins 0.000 claims description 3
- 102000014150 Interferons Human genes 0.000 claims description 3
- 108010050904 Interferons Proteins 0.000 claims description 3
- 102000003810 Interleukin-18 Human genes 0.000 claims description 3
- 108090000171 Interleukin-18 Proteins 0.000 claims description 3
- 108010002616 Interleukin-5 Proteins 0.000 claims description 3
- 102000007547 Laminin Human genes 0.000 claims description 3
- 108010085895 Laminin Proteins 0.000 claims description 3
- 241000713666 Lentivirus Species 0.000 claims description 3
- 108060003100 Magainin Proteins 0.000 claims description 3
- 102000000424 Matrix Metalloproteinase 2 Human genes 0.000 claims description 3
- 108010016165 Matrix Metalloproteinase 2 Proteins 0.000 claims description 3
- 108010036176 Melitten Proteins 0.000 claims description 3
- 101000686934 Mus musculus Prolactin-7D1 Proteins 0.000 claims description 3
- MSHZHSPISPJWHW-PVDLLORBSA-N O-(chloroacetylcarbamoyl)fumagillol Chemical compound C([C@H]([C@H]([C@@H]1[C@]2(C)[C@H](O2)CC=C(C)C)OC)OC(=O)NC(=O)CCl)C[C@@]21CO2 MSHZHSPISPJWHW-PVDLLORBSA-N 0.000 claims description 3
- BYPFEZZEUUWMEJ-UHFFFAOYSA-N Pentoxifylline Chemical compound O=C1N(CCCCC(=O)C)C(=O)N(C)C2=C1N(C)C=N2 BYPFEZZEUUWMEJ-UHFFFAOYSA-N 0.000 claims description 3
- 102100035846 Pigment epithelium-derived factor Human genes 0.000 claims description 3
- 102000010752 Plasminogen Inactivators Human genes 0.000 claims description 3
- 108010077971 Plasminogen Inactivators Proteins 0.000 claims description 3
- 102000003946 Prolactin Human genes 0.000 claims description 3
- 108010057464 Prolactin Proteins 0.000 claims description 3
- 229940079156 Proteasome inhibitor Drugs 0.000 claims description 3
- 102400001051 Restin Human genes 0.000 claims description 3
- 101800000689 Restin Proteins 0.000 claims description 3
- 206010039491 Sarcoma Diseases 0.000 claims description 3
- 108010041979 accutin Proteins 0.000 claims description 3
- 230000002424 anti-apoptotic effect Effects 0.000 claims description 3
- 230000003115 biocidal effect Effects 0.000 claims description 3
- 210000000481 breast Anatomy 0.000 claims description 3
- 210000004899 c-terminal region Anatomy 0.000 claims description 3
- 229960000724 cidofovir Drugs 0.000 claims description 3
- 208000035475 disorder Diseases 0.000 claims description 3
- 229940045109 genistein Drugs 0.000 claims description 3
- TZBJGXHYKVUXJN-UHFFFAOYSA-N genistein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O TZBJGXHYKVUXJN-UHFFFAOYSA-N 0.000 claims description 3
- 235000006539 genistein Nutrition 0.000 claims description 3
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 claims description 3
- 229960004905 gramicidin Drugs 0.000 claims description 3
- ZWCXYZRRTRDGQE-SORVKSEFSA-N gramicidina Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](C(C)C)NC(=O)[C@H](C)NC(=O)[C@H](NC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](NC=O)C(C)C)CC(C)C)C(=O)NCCO)=CNC2=C1 ZWCXYZRRTRDGQE-SORVKSEFSA-N 0.000 claims description 3
- MCAHMSDENAOJFZ-BVXDHVRPSA-N herbimycin Chemical compound N1C(=O)\C(C)=C\C=C/[C@H](OC)[C@@H](OC(N)=O)\C(C)=C\[C@H](C)[C@@H](OC)[C@@H](OC)C[C@H](C)[C@@H](OC)C2=CC(=O)C=C1C2=O MCAHMSDENAOJFZ-BVXDHVRPSA-N 0.000 claims description 3
- 239000003667 hormone antagonist Substances 0.000 claims description 3
- 229960003130 interferon gamma Drugs 0.000 claims description 3
- 229940047124 interferons Drugs 0.000 claims description 3
- 229940117681 interleukin-12 Drugs 0.000 claims description 3
- 229940043355 kinase inhibitor Drugs 0.000 claims description 3
- 230000005291 magnetic effect Effects 0.000 claims description 3
- OCSMOTCMPXTDND-OUAUKWLOSA-N marimastat Chemical compound CNC(=O)[C@H](C(C)(C)C)NC(=O)[C@H](CC(C)C)[C@H](O)C(=O)NO OCSMOTCMPXTDND-OUAUKWLOSA-N 0.000 claims description 3
- 229950008959 marimastat Drugs 0.000 claims description 3
- VDXZNPDIRNWWCW-UHFFFAOYSA-N melitten Chemical compound NCC(=O)NC(C(C)CC)C(=O)NCC(=O)NC(C)C(=O)NC(C(C)C)C(=O)NC(CC(C)C)C(=O)NC(CCCCN)C(=O)NC(C(C)C)C(=O)NC(CC(C)C)C(=O)NC(C(C)O)C(=O)NC(C(C)O)C(=O)NCC(=O)NC(CC(C)C)C(=O)N1CCCC1C(=O)NC(C)C(=O)NC(CC(C)C)C(=O)NC(C(C)CC)C(=O)NC(CO)C(=O)NC(C(=O)NC(C(C)CC)C(=O)NC(CCCCN)C(=O)NC(CCCNC(N)=N)C(=O)NC(CCCCN)C(=O)NC(CCCNC(N)=N)C(=O)NC(CCC(N)=O)C(=O)NC(CCC(N)=O)C(N)=O)CC1=CNC2=CC=CC=C12 VDXZNPDIRNWWCW-UHFFFAOYSA-N 0.000 claims description 3
- 239000003475 metalloproteinase inhibitor Substances 0.000 claims description 3
- 239000011859 microparticle Substances 0.000 claims description 3
- DYKFCLLONBREIL-KVUCHLLUSA-N minocycline Chemical compound C([C@H]1C2)C3=C(N(C)C)C=CC(O)=C3C(=O)C1=C(O)[C@@]1(O)[C@@H]2[C@H](N(C)C)C(O)=C(C(N)=O)C1=O DYKFCLLONBREIL-KVUCHLLUSA-N 0.000 claims description 3
- 229960004023 minocycline Drugs 0.000 claims description 3
- 230000002611 ovarian Effects 0.000 claims description 3
- 229960001476 pentoxifylline Drugs 0.000 claims description 3
- 239000003757 phosphotransferase inhibitor Substances 0.000 claims description 3
- 108090000102 pigment epithelium-derived factor Proteins 0.000 claims description 3
- 239000002797 plasminogen activator inhibitor Substances 0.000 claims description 3
- 229920000768 polyamine Polymers 0.000 claims description 3
- 229940097325 prolactin Drugs 0.000 claims description 3
- 239000003207 proteasome inhibitor Substances 0.000 claims description 3
- 229960003522 roquinimex Drugs 0.000 claims description 3
- 210000003491 skin Anatomy 0.000 claims description 3
- 229960003433 thalidomide Drugs 0.000 claims description 3
- 210000005253 yeast cell Anatomy 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 241000702421 Dependoparvovirus Species 0.000 claims description 2
- 108090000174 Interleukin-10 Proteins 0.000 claims description 2
- 206010025323 Lymphomas Diseases 0.000 claims description 2
- 206010029260 Neuroblastoma Diseases 0.000 claims description 2
- MSHZHSPISPJWHW-UHFFFAOYSA-N O-(chloroacetylcarbamoyl)fumagillol Chemical compound O1C(CC=C(C)C)C1(C)C1C(OC)C(OC(=O)NC(=O)CCl)CCC21CO2 MSHZHSPISPJWHW-UHFFFAOYSA-N 0.000 claims description 2
- 210000000988 bone and bone Anatomy 0.000 claims description 2
- 210000001072 colon Anatomy 0.000 claims description 2
- 210000001508 eye Anatomy 0.000 claims description 2
- 230000002496 gastric effect Effects 0.000 claims description 2
- 210000003734 kidney Anatomy 0.000 claims description 2
- 208000032839 leukemia Diseases 0.000 claims description 2
- 210000004185 liver Anatomy 0.000 claims description 2
- 210000004072 lung Anatomy 0.000 claims description 2
- 210000002307 prostate Anatomy 0.000 claims description 2
- 230000012743 protein tagging Effects 0.000 claims description 2
- 230000002381 testicular Effects 0.000 claims description 2
- 210000002105 tongue Anatomy 0.000 claims description 2
- 210000003932 urinary bladder Anatomy 0.000 claims description 2
- 102000009075 Angiopoietin-2 Human genes 0.000 claims 1
- 238000011282 treatment Methods 0.000 abstract description 27
- 150000001875 compounds Chemical class 0.000 abstract description 14
- 230000000973 chemotherapeutic effect Effects 0.000 abstract description 7
- 230000004927 fusion Effects 0.000 abstract description 6
- 235000018102 proteins Nutrition 0.000 description 107
- 239000013598 vector Substances 0.000 description 29
- 235000001014 amino acid Nutrition 0.000 description 27
- 108020004414 DNA Proteins 0.000 description 25
- 229940024606 amino acid Drugs 0.000 description 25
- 239000003446 ligand Substances 0.000 description 22
- 230000014509 gene expression Effects 0.000 description 21
- 241000699670 Mus sp. Species 0.000 description 19
- 210000004881 tumor cell Anatomy 0.000 description 19
- 108020004459 Small interfering RNA Proteins 0.000 description 17
- 230000000694 effects Effects 0.000 description 17
- 230000037361 pathway Effects 0.000 description 16
- 230000001225 therapeutic effect Effects 0.000 description 16
- 238000001415 gene therapy Methods 0.000 description 15
- -1 IL-IO Proteins 0.000 description 14
- 230000001404 mediated effect Effects 0.000 description 14
- 229920001184 polypeptide Polymers 0.000 description 14
- 239000000126 substance Substances 0.000 description 14
- 238000001727 in vivo Methods 0.000 description 13
- 239000003112 inhibitor Substances 0.000 description 13
- 150000007523 nucleic acids Chemical class 0.000 description 13
- 238000000746 purification Methods 0.000 description 13
- 239000004055 small Interfering RNA Substances 0.000 description 13
- 230000003834 intracellular effect Effects 0.000 description 12
- 102000039446 nucleic acids Human genes 0.000 description 12
- 108020004707 nucleic acids Proteins 0.000 description 12
- 230000000861 pro-apoptotic effect Effects 0.000 description 12
- 102000005962 receptors Human genes 0.000 description 12
- 108020003175 receptors Proteins 0.000 description 12
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 11
- 230000004807 localization Effects 0.000 description 11
- 239000013642 negative control Substances 0.000 description 11
- WEVYNIUIFUYDGI-UHFFFAOYSA-N 3-[6-[4-(trifluoromethoxy)anilino]-4-pyrimidinyl]benzamide Chemical compound NC(=O)C1=CC=CC(C=2N=CN=C(NC=3C=CC(OC(F)(F)F)=CC=3)C=2)=C1 WEVYNIUIFUYDGI-UHFFFAOYSA-N 0.000 description 10
- 108091054455 MAP kinase family Proteins 0.000 description 10
- 102000043136 MAP kinase family Human genes 0.000 description 10
- 102000000395 SH3 domains Human genes 0.000 description 10
- 108050008861 SH3 domains Proteins 0.000 description 10
- 230000004663 cell proliferation Effects 0.000 description 10
- 239000003431 cross linking reagent Substances 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 10
- 230000006870 function Effects 0.000 description 10
- 210000000056 organ Anatomy 0.000 description 10
- 125000003275 alpha amino acid group Chemical group 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 230000003993 interaction Effects 0.000 description 9
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 8
- 230000012292 cell migration Effects 0.000 description 8
- 125000004122 cyclic group Chemical group 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 229940124597 therapeutic agent Drugs 0.000 description 8
- 238000012546 transfer Methods 0.000 description 8
- 108020004635 Complementary DNA Proteins 0.000 description 7
- 101100015729 Drosophila melanogaster drk gene Proteins 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 7
- 108091034117 Oligonucleotide Proteins 0.000 description 7
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 7
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 7
- 238000001042 affinity chromatography Methods 0.000 description 7
- 210000000170 cell membrane Anatomy 0.000 description 7
- 101150098203 grb2 gene Proteins 0.000 description 7
- 230000003463 hyperproliferative effect Effects 0.000 description 7
- 238000003384 imaging method Methods 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- 238000002823 phage display Methods 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 238000000159 protein binding assay Methods 0.000 description 7
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 230000005754 cellular signaling Effects 0.000 description 6
- 238000002512 chemotherapy Methods 0.000 description 6
- 238000004132 cross linking Methods 0.000 description 6
- 230000001472 cytotoxic effect Effects 0.000 description 6
- 230000001419 dependent effect Effects 0.000 description 6
- 125000000524 functional group Chemical group 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 238000002372 labelling Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 238000013508 migration Methods 0.000 description 6
- 102000040430 polynucleotide Human genes 0.000 description 6
- 108091033319 polynucleotide Proteins 0.000 description 6
- 239000002157 polynucleotide Substances 0.000 description 6
- 238000001959 radiotherapy Methods 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 241000699660 Mus musculus Species 0.000 description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- 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 5
- 230000004913 activation Effects 0.000 description 5
- 229940100198 alkylating agent Drugs 0.000 description 5
- 239000002168 alkylating agent Substances 0.000 description 5
- 125000000539 amino acid group Chemical group 0.000 description 5
- 230000000340 anti-metabolite Effects 0.000 description 5
- 229940100197 antimetabolite Drugs 0.000 description 5
- 239000002256 antimetabolite Substances 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- 238000010804 cDNA synthesis Methods 0.000 description 5
- 230000021164 cell adhesion Effects 0.000 description 5
- 230000022131 cell cycle Effects 0.000 description 5
- 230000030833 cell death Effects 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 231100000433 cytotoxic Toxicity 0.000 description 5
- 239000013604 expression vector Substances 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- 239000000017 hydrogel Substances 0.000 description 5
- 238000001114 immunoprecipitation Methods 0.000 description 5
- 239000011159 matrix material Substances 0.000 description 5
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical compound ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 5
- 238000011580 nude mouse model Methods 0.000 description 5
- 238000004806 packaging method and process Methods 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- 239000000816 peptidomimetic Substances 0.000 description 5
- 239000012071 phase Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 102000016914 ras Proteins Human genes 0.000 description 5
- 108010014186 ras Proteins Proteins 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 239000012679 serum free medium Substances 0.000 description 5
- 238000001356 surgical procedure Methods 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical class N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 description 4
- 108700026790 CRKL Proteins 0.000 description 4
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 4
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 4
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 102000001702 Intracellular Signaling Peptides and Proteins Human genes 0.000 description 4
- 108010068964 Intracellular Signaling Peptides and Proteins Proteins 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 4
- 229930040373 Paraformaldehyde Natural products 0.000 description 4
- 108010067902 Peptide Library Proteins 0.000 description 4
- 102000014400 SH2 domains Human genes 0.000 description 4
- 108050003452 SH2 domains Proteins 0.000 description 4
- NKANXQFJJICGDU-QPLCGJKRSA-N Tamoxifen Chemical compound C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 NKANXQFJJICGDU-QPLCGJKRSA-N 0.000 description 4
- 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 description 4
- 230000003213 activating effect Effects 0.000 description 4
- 102000035181 adaptor proteins Human genes 0.000 description 4
- 108091005764 adaptor proteins Proteins 0.000 description 4
- 229940088710 antibiotic agent Drugs 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 4
- 230000001588 bifunctional effect Effects 0.000 description 4
- 239000011230 binding agent Substances 0.000 description 4
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 4
- 229960004316 cisplatin Drugs 0.000 description 4
- 238000002648 combination therapy Methods 0.000 description 4
- 239000000824 cytostatic agent Substances 0.000 description 4
- 230000001085 cytostatic effect Effects 0.000 description 4
- 239000012636 effector Substances 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 229960002949 fluorouracil Drugs 0.000 description 4
- 238000005194 fractionation Methods 0.000 description 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 4
- 229910052737 gold Inorganic materials 0.000 description 4
- 239000010931 gold Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- 230000000670 limiting effect Effects 0.000 description 4
- 229960004961 mechlorethamine Drugs 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 230000000394 mitotic effect Effects 0.000 description 4
- 238000010172 mouse model Methods 0.000 description 4
- 239000002105 nanoparticle Substances 0.000 description 4
- 229920002866 paraformaldehyde Polymers 0.000 description 4
- 239000008194 pharmaceutical composition Substances 0.000 description 4
- 230000000144 pharmacologic effect Effects 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 238000007639 printing Methods 0.000 description 4
- CPTBDICYNRMXFX-UHFFFAOYSA-N procarbazine Chemical compound CNNCC1=CC=C(C(=O)NC(C)C)C=C1 CPTBDICYNRMXFX-UHFFFAOYSA-N 0.000 description 4
- 238000001742 protein purification Methods 0.000 description 4
- 230000004850 protein–protein interaction Effects 0.000 description 4
- 230000001177 retroviral effect Effects 0.000 description 4
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 description 4
- 230000019491 signal transduction Effects 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 238000004627 transmission electron microscopy Methods 0.000 description 4
- 230000004614 tumor growth Effects 0.000 description 4
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 4
- 229960003048 vinblastine Drugs 0.000 description 4
- 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 description 4
- 238000001262 western blot Methods 0.000 description 4
- 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 description 3
- 102000005416 ATP-Binding Cassette Transporters Human genes 0.000 description 3
- 108010006533 ATP-Binding Cassette Transporters Proteins 0.000 description 3
- 108090000672 Annexin A5 Proteins 0.000 description 3
- 102000004121 Annexin A5 Human genes 0.000 description 3
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 3
- 102000000844 Cell Surface Receptors Human genes 0.000 description 3
- 108010001857 Cell Surface Receptors Proteins 0.000 description 3
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 description 3
- 108010092160 Dactinomycin Proteins 0.000 description 3
- 241000287828 Gallus gallus Species 0.000 description 3
- 101000606500 Gallus gallus Inactive tyrosine-protein kinase 7 Proteins 0.000 description 3
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 3
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 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 description 3
- 108010052285 Membrane Proteins Proteins 0.000 description 3
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 description 3
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 3
- 241001028048 Nicola Species 0.000 description 3
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 3
- 102000012005 alpha-2-HS-Glycoprotein Human genes 0.000 description 3
- 108010075843 alpha-2-HS-Glycoprotein Proteins 0.000 description 3
- QWCKQJZIFLGMSD-UHFFFAOYSA-N alpha-aminobutyric acid Chemical compound CCC(N)C(O)=O QWCKQJZIFLGMSD-UHFFFAOYSA-N 0.000 description 3
- 230000000692 anti-sense effect Effects 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 238000003782 apoptosis assay Methods 0.000 description 3
- VSRXQHXAPYXROS-UHFFFAOYSA-N azanide;cyclobutane-1,1-dicarboxylic acid;platinum(2+) Chemical compound [NH2-].[NH2-].[Pt+2].OC(=O)C1(C(O)=O)CCC1 VSRXQHXAPYXROS-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 229960002092 busulfan Drugs 0.000 description 3
- 229960004562 carboplatin Drugs 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 230000022534 cell killing Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 229960004630 chlorambucil Drugs 0.000 description 3
- JCKYGMPEJWAADB-UHFFFAOYSA-N chlorambucil Chemical compound OC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 JCKYGMPEJWAADB-UHFFFAOYSA-N 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 238000011498 curative surgery Methods 0.000 description 3
- 229960004397 cyclophosphamide Drugs 0.000 description 3
- 229960000640 dactinomycin Drugs 0.000 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 description 3
- 229960000975 daunorubicin Drugs 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 102000015694 estrogen receptors Human genes 0.000 description 3
- 108010038795 estrogen receptors Proteins 0.000 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 description 3
- 238000009472 formulation Methods 0.000 description 3
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 3
- 229960001101 ifosfamide Drugs 0.000 description 3
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 238000009169 immunotherapy Methods 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229960001924 melphalan Drugs 0.000 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 description 3
- 229960000485 methotrexate Drugs 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- HDZGCSFEDULWCS-UHFFFAOYSA-N monomethylhydrazine Chemical class CNN HDZGCSFEDULWCS-UHFFFAOYSA-N 0.000 description 3
- 229930014626 natural product Natural products 0.000 description 3
- 238000011275 oncology therapy Methods 0.000 description 3
- 229960003330 pentetic acid Drugs 0.000 description 3
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 229960000624 procarbazine Drugs 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000005522 programmed cell death Effects 0.000 description 3
- 238000002271 resection Methods 0.000 description 3
- 239000004017 serum-free culture medium Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 2
- UWFRVQVNYNPBEF-UHFFFAOYSA-N 1-(2,4-dimethylphenyl)propan-1-one Chemical compound CCC(=O)C1=CC=C(C)C=C1C UWFRVQVNYNPBEF-UHFFFAOYSA-N 0.000 description 2
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 2
- FUOOLUPWFVMBKG-UHFFFAOYSA-N 2-Aminoisobutyric acid Chemical compound CC(C)(N)C(O)=O FUOOLUPWFVMBKG-UHFFFAOYSA-N 0.000 description 2
- OYIFNHCXNCRBQI-UHFFFAOYSA-N 2-aminoadipic acid Chemical compound OC(=O)C(N)CCCC(O)=O OYIFNHCXNCRBQI-UHFFFAOYSA-N 0.000 description 2
- RDFMDVXONNIGBC-UHFFFAOYSA-N 2-aminoheptanoic acid Chemical compound CCCCCC(N)C(O)=O RDFMDVXONNIGBC-UHFFFAOYSA-N 0.000 description 2
- SNDPXSYFESPGGJ-UHFFFAOYSA-N 2-aminopentanoic acid Chemical compound CCCC(N)C(O)=O SNDPXSYFESPGGJ-UHFFFAOYSA-N 0.000 description 2
- MEOVPKDOYAIVHZ-UHFFFAOYSA-N 2-chloro-1-(1-methylpyrrol-2-yl)ethanol Chemical compound CN1C=CC=C1C(O)CCl MEOVPKDOYAIVHZ-UHFFFAOYSA-N 0.000 description 2
- PECYZEOJVXMISF-UHFFFAOYSA-N 3-aminoalanine Chemical compound [NH3+]CC(N)C([O-])=O PECYZEOJVXMISF-UHFFFAOYSA-N 0.000 description 2
- BYXHQQCXAJARLQ-ZLUOBGJFSA-N Ala-Ala-Ala Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O BYXHQQCXAJARLQ-ZLUOBGJFSA-N 0.000 description 2
- 102100034608 Angiopoietin-2 Human genes 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 101000583080 Bunodosoma granuliferum Delta-actitoxin-Bgr2a Proteins 0.000 description 2
- 102100025248 C-X-C motif chemokine 10 Human genes 0.000 description 2
- 101710098275 C-X-C motif chemokine 10 Proteins 0.000 description 2
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 239000004971 Cross linker Substances 0.000 description 2
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 241000275449 Diplectrum formosum Species 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 102000016621 Focal Adhesion Protein-Tyrosine Kinases Human genes 0.000 description 2
- 108010067715 Focal Adhesion Protein-Tyrosine Kinases Proteins 0.000 description 2
- 229910052688 Gadolinium Inorganic materials 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- 208000007766 Kaposi sarcoma Diseases 0.000 description 2
- JUQLUIFNNFIIKC-YFKPBYRVSA-N L-2-aminopimelic acid Chemical compound OC(=O)[C@@H](N)CCCCC(O)=O JUQLUIFNNFIIKC-YFKPBYRVSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 101150024075 Mapk1 gene Proteins 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 229930192392 Mitomycin Natural products 0.000 description 2
- 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 description 2
- HRNLUBSXIHFDHP-UHFFFAOYSA-N N-(2-aminophenyl)-4-[[[4-(3-pyridinyl)-2-pyrimidinyl]amino]methyl]benzamide Chemical compound NC1=CC=CC=C1NC(=O)C(C=C1)=CC=C1CNC1=NC=CC(C=2C=NC=CC=2)=N1 HRNLUBSXIHFDHP-UHFFFAOYSA-N 0.000 description 2
- KSPIYJQBLVDRRI-UHFFFAOYSA-N N-methylisoleucine Chemical compound CCC(C)C(NC)C(O)=O KSPIYJQBLVDRRI-UHFFFAOYSA-N 0.000 description 2
- 206010029113 Neovascularisation Diseases 0.000 description 2
- 108700020796 Oncogene Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 108091000080 Phosphotransferase Proteins 0.000 description 2
- 101710086988 Protein terminus Proteins 0.000 description 2
- 108010090931 Proto-Oncogene Proteins c-bcl-2 Proteins 0.000 description 2
- 102000013535 Proto-Oncogene Proteins c-bcl-2 Human genes 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- 108010077895 Sarcosine Proteins 0.000 description 2
- 102000039471 Small Nuclear RNA Human genes 0.000 description 2
- 108010090804 Streptavidin Chemical class 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- HATRDXDCPOXQJX-UHFFFAOYSA-N Thapsigargin Natural products CCCCCCCC(=O)OC1C(OC(O)C(=C/C)C)C(=C2C3OC(=O)C(C)(O)C3(O)C(CC(C)(OC(=O)C)C12)OC(=O)CCC)C HATRDXDCPOXQJX-UHFFFAOYSA-N 0.000 description 2
- 102000016548 Vascular Endothelial Growth Factor Receptor-1 Human genes 0.000 description 2
- 108010053096 Vascular Endothelial Growth Factor Receptor-1 Proteins 0.000 description 2
- 229940122803 Vinca alkaloid Drugs 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000009056 active transport Effects 0.000 description 2
- 102000019997 adhesion receptor Human genes 0.000 description 2
- 108010013985 adhesion receptor Proteins 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 238000001261 affinity purification Methods 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 2
- 150000001412 amines Chemical group 0.000 description 2
- RGHILYZRVFRRNK-UHFFFAOYSA-N anthracene-1,2-dione Chemical class C1=CC=C2C=C(C(C(=O)C=C3)=O)C3=CC2=C1 RGHILYZRVFRRNK-UHFFFAOYSA-N 0.000 description 2
- 239000003429 antifungal agent Substances 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000003305 autocrine Effects 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- KQNZDYYTLMIZCT-KQPMLPITSA-N brefeldin A Chemical compound O[C@@H]1\C=C\C(=O)O[C@@H](C)CCC\C=C\[C@@H]2C[C@H](O)C[C@H]21 KQNZDYYTLMIZCT-KQPMLPITSA-N 0.000 description 2
- JUMGSHROWPPKFX-UHFFFAOYSA-N brefeldin-A Natural products CC1CCCC=CC2(C)CC(O)CC2(C)C(O)C=CC(=O)O1 JUMGSHROWPPKFX-UHFFFAOYSA-N 0.000 description 2
- 229940127093 camptothecin Drugs 0.000 description 2
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 235000013877 carbamide Nutrition 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 231100000504 carcinogenesis Toxicity 0.000 description 2
- 238000001516 cell proliferation assay Methods 0.000 description 2
- 230000015861 cell surface binding Effects 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- OSASVXMJTNOKOY-UHFFFAOYSA-N chlorobutanol Chemical compound CC(C)(O)C(Cl)(Cl)Cl OSASVXMJTNOKOY-UHFFFAOYSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 229910017052 cobalt Inorganic materials 0.000 description 2
- 239000010941 cobalt Substances 0.000 description 2
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 239000007822 coupling agent Substances 0.000 description 2
- 230000009260 cross reactivity Effects 0.000 description 2
- 231100000409 cytocidal Toxicity 0.000 description 2
- 230000000445 cytocidal effect Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- VEVRNHHLCPGNDU-MUGJNUQGSA-O desmosine Chemical compound OC(=O)[C@@H](N)CCCC[N+]1=CC(CC[C@H](N)C(O)=O)=C(CCC[C@H](N)C(O)=O)C(CC[C@H](N)C(O)=O)=C1 VEVRNHHLCPGNDU-MUGJNUQGSA-O 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 229960001484 edetic acid Drugs 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 description 2
- 210000003976 gap junction Anatomy 0.000 description 2
- 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 description 2
- 229960005277 gemcitabine Drugs 0.000 description 2
- ZNNLBTZKUZBEKO-UHFFFAOYSA-N glyburide Chemical compound COC1=CC=C(Cl)C=C1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCCCC2)C=C1 ZNNLBTZKUZBEKO-UHFFFAOYSA-N 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- 238000001794 hormone therapy Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000002519 immonomodulatory effect Effects 0.000 description 2
- 239000012642 immune effector Substances 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 229940121354 immunomodulator Drugs 0.000 description 2
- 230000001024 immunotherapeutic effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- APFVFJFRJDLVQX-AHCXROLUSA-N indium-111 Chemical compound [111In] APFVFJFRJDLVQX-AHCXROLUSA-N 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 238000004255 ion exchange chromatography Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- RGXCTRIQQODGIZ-UHFFFAOYSA-O isodesmosine Chemical compound OC(=O)C(N)CCCC[N+]1=CC(CCC(N)C(O)=O)=CC(CCC(N)C(O)=O)=C1CCCC(N)C(O)=O RGXCTRIQQODGIZ-UHFFFAOYSA-O 0.000 description 2
- 238000001155 isoelectric focusing Methods 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 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 description 2
- 239000003226 mitogen Substances 0.000 description 2
- 229960004857 mitomycin Drugs 0.000 description 2
- 230000011278 mitosis Effects 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 230000000869 mutational effect Effects 0.000 description 2
- 229940086322 navelbine Drugs 0.000 description 2
- 229910052759 nickel Inorganic materials 0.000 description 2
- 239000007800 oxidant agent Substances 0.000 description 2
- 230000003076 paracrine Effects 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 230000010412 perfusion Effects 0.000 description 2
- 108091005981 phosphorylated proteins Proteins 0.000 description 2
- 102000020233 phosphotransferase Human genes 0.000 description 2
- 229910052697 platinum Inorganic materials 0.000 description 2
- 229960003171 plicamycin Drugs 0.000 description 2
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 201000001514 prostate carcinoma Diseases 0.000 description 2
- 208000023958 prostate neoplasm Diseases 0.000 description 2
- 230000006916 protein interaction Effects 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 230000003439 radiotherapeutic effect Effects 0.000 description 2
- 229960004622 raloxifene Drugs 0.000 description 2
- GZUITABIAKMVPG-UHFFFAOYSA-N raloxifene Chemical compound C1=CC(O)=CC=C1C1=C(C(=O)C=2C=CC(OCCN3CCCCC3)=CC=2)C2=CC=C(O)C=C2S1 GZUITABIAKMVPG-UHFFFAOYSA-N 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 108091029842 small nuclear ribonucleic acid Proteins 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- FVAUCKIRQBBSSJ-UHFFFAOYSA-M sodium iodide Inorganic materials [Na+].[I-] FVAUCKIRQBBSSJ-UHFFFAOYSA-M 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 231100000057 systemic toxicity Toxicity 0.000 description 2
- 229960001603 tamoxifen Drugs 0.000 description 2
- GKLVYJBZJHMRIY-UHFFFAOYSA-N technetium atom Chemical compound [Tc] GKLVYJBZJHMRIY-UHFFFAOYSA-N 0.000 description 2
- IXFPJGBNCFXKPI-FSIHEZPISA-N thapsigargin Chemical compound CCCC(=O)O[C@H]1C[C@](C)(OC(C)=O)[C@H]2[C@H](OC(=O)CCCCCCC)[C@@H](OC(=O)C(\C)=C/C)C(C)=C2[C@@H]2OC(=O)[C@@](C)(O)[C@]21O IXFPJGBNCFXKPI-FSIHEZPISA-N 0.000 description 2
- 230000026683 transduction Effects 0.000 description 2
- 238000010361 transduction Methods 0.000 description 2
- 102000027257 transmembrane receptors Human genes 0.000 description 2
- 108091008578 transmembrane receptors Proteins 0.000 description 2
- 238000013414 tumor xenograft model Methods 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- 150000003672 ureas Chemical class 0.000 description 2
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 2
- 210000005166 vasculature Anatomy 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- CILBMBUYJCWATM-PYGJLNRPSA-N vinorelbine ditartrate Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O.OC(=O)[C@H](O)[C@@H](O)C(O)=O.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 CILBMBUYJCWATM-PYGJLNRPSA-N 0.000 description 2
- 239000013603 viral vector Substances 0.000 description 2
- ATYCFNRXENKXSE-MHPIHPPYSA-N (2,5-dioxopyrrolidin-1-yl) 6-[6-[5-[(3as,4s,6ar)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]hexanoylamino]hexanoate Chemical compound C([C@H]1[C@H]2NC(=O)N[C@H]2CS1)CCCC(=O)NCCCCCC(=O)NCCCCCC(=O)ON1C(=O)CCC1=O ATYCFNRXENKXSE-MHPIHPPYSA-N 0.000 description 1
- BJBUEDPLEOHJGE-UHFFFAOYSA-N (2R,3S)-3-Hydroxy-2-pyrolidinecarboxylic acid Natural products OC1CCNC1C(O)=O BJBUEDPLEOHJGE-UHFFFAOYSA-N 0.000 description 1
- MWWSFMDVAYGXBV-MYPASOLCSA-N (7r,9s)-7-[(2r,4s,5s,6s)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound Cl.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 MWWSFMDVAYGXBV-MYPASOLCSA-N 0.000 description 1
- JHTPBGFVWWSHDL-UHFFFAOYSA-N 1,4-dichloro-2-isothiocyanatobenzene Chemical compound ClC1=CC=C(Cl)C(N=C=S)=C1 JHTPBGFVWWSHDL-UHFFFAOYSA-N 0.000 description 1
- 102100025573 1-alkyl-2-acetylglycerophosphocholine esterase Human genes 0.000 description 1
- PNDPGZBMCMUPRI-HVTJNCQCSA-N 10043-66-0 Chemical compound [131I][131I] PNDPGZBMCMUPRI-HVTJNCQCSA-N 0.000 description 1
- WUAPFZMCVAUBPE-NJFSPNSNSA-N 188Re Chemical compound [188Re] WUAPFZMCVAUBPE-NJFSPNSNSA-N 0.000 description 1
- OGNSCSPNOLGXSM-UHFFFAOYSA-N 2,4-diaminobutyric acid Chemical compound NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 description 1
- GMKMEZVLHJARHF-UHFFFAOYSA-N 2,6-diaminopimelic acid Chemical compound OC(=O)C(N)CCCC(N)C(O)=O GMKMEZVLHJARHF-UHFFFAOYSA-N 0.000 description 1
- QZWBOYMQPQVGPM-UHFFFAOYSA-N 2-(1h-indol-2-yl)guanidine Chemical compound C1=CC=C2NC(NC(=N)N)=CC2=C1 QZWBOYMQPQVGPM-UHFFFAOYSA-N 0.000 description 1
- CTRPRMNBTVRDFH-UHFFFAOYSA-N 2-n-methyl-1,3,5-triazine-2,4,6-triamine Chemical compound CNC1=NC(N)=NC(N)=N1 CTRPRMNBTVRDFH-UHFFFAOYSA-N 0.000 description 1
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- PECYZEOJVXMISF-REOHCLBHSA-N 3-amino-L-alanine Chemical compound [NH3+]C[C@H](N)C([O-])=O PECYZEOJVXMISF-REOHCLBHSA-N 0.000 description 1
- XABCFXXGZPWJQP-UHFFFAOYSA-N 3-aminoadipic acid Chemical compound OC(=O)CC(N)CCC(O)=O XABCFXXGZPWJQP-UHFFFAOYSA-N 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 101710134681 40 kDa protein Proteins 0.000 description 1
- SLXKOJJOQWFEFD-UHFFFAOYSA-N 6-aminohexanoic acid Chemical compound NCCCCCC(O)=O SLXKOJJOQWFEFD-UHFFFAOYSA-N 0.000 description 1
- YYSFXUWWPNHNAZ-OSDRTFJJSA-N 851536-75-9 Chemical compound C1[C@@H](OC)[C@H](OCCOCC)CC[C@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CCC2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 YYSFXUWWPNHNAZ-OSDRTFJJSA-N 0.000 description 1
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 1
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 1
- 108700026758 Adenovirus hexon capsid Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108020004491 Antisense DNA Proteins 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 108091023037 Aptamer Proteins 0.000 description 1
- 240000003291 Armoracia rusticana Species 0.000 description 1
- 235000011330 Armoracia rusticana Nutrition 0.000 description 1
- 108010024976 Asparaginase Chemical class 0.000 description 1
- 108090001008 Avidin Chemical class 0.000 description 1
- 102100035634 B-cell linker protein Human genes 0.000 description 1
- 101710083670 B-cell linker protein Proteins 0.000 description 1
- 241000005672 Baia Species 0.000 description 1
- 102000051485 Bcl-2 family Human genes 0.000 description 1
- 108700038897 Bcl-2 family Proteins 0.000 description 1
- 241000701822 Bovine papillomavirus Species 0.000 description 1
- 238000007809 Boyden Chamber assay Methods 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 description 1
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 102000001327 Chemokine CCL5 Human genes 0.000 description 1
- 108010055166 Chemokine CCL5 Proteins 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 241000557626 Corvus corax Species 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
- 102000010831 Cytoskeletal Proteins Human genes 0.000 description 1
- 108010037414 Cytoskeletal Proteins Proteins 0.000 description 1
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 241000450599 DNA viruses Species 0.000 description 1
- 101100347633 Drosophila melanogaster Mhc gene Proteins 0.000 description 1
- 229910052692 Dysprosium Inorganic materials 0.000 description 1
- 102400001368 Epidermal growth factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 229910052691 Erbium Inorganic materials 0.000 description 1
- 101001039702 Escherichia coli (strain K12) Methyl-accepting chemotaxis protein I Proteins 0.000 description 1
- 102100029951 Estrogen receptor beta Human genes 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- 101710145505 Fiber protein Proteins 0.000 description 1
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 1
- GYHNNYVSQQEPJS-OIOBTWANSA-N Gallium-67 Chemical compound [67Ga] GYHNNYVSQQEPJS-OIOBTWANSA-N 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 102100033067 Growth factor receptor-bound protein 2 Human genes 0.000 description 1
- 108091009389 Growth factor receptor-bound protein 2 Proteins 0.000 description 1
- 208000002250 Hematologic Neoplasms Diseases 0.000 description 1
- 208000009889 Herpes Simplex Diseases 0.000 description 1
- 101001010910 Homo sapiens Estrogen receptor beta Proteins 0.000 description 1
- 101001034314 Homo sapiens Lactadherin Proteins 0.000 description 1
- LCWXJXMHJVIJFK-UHFFFAOYSA-N Hydroxylysine Natural products NCC(O)CC(N)CC(O)=O LCWXJXMHJVIJFK-UHFFFAOYSA-N 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 1
- VSNHCAURESNICA-UHFFFAOYSA-N Hydroxyurea Chemical compound NC(=O)NO VSNHCAURESNICA-UHFFFAOYSA-N 0.000 description 1
- XDXDZDZNSLXDNA-TZNDIEGXSA-N Idarubicin Chemical compound C1[C@H](N)[C@H](O)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2C[C@@](O)(C(C)=O)C1 XDXDZDZNSLXDNA-TZNDIEGXSA-N 0.000 description 1
- XDXDZDZNSLXDNA-UHFFFAOYSA-N Idarubicin Natural products C1C(N)C(O)C(C)OC1OC1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2CC(O)(C(C)=O)C1 XDXDZDZNSLXDNA-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 102000003996 Interferon-beta Human genes 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- 102000003815 Interleukin-11 Human genes 0.000 description 1
- 108090000177 Interleukin-11 Proteins 0.000 description 1
- ZCYVEMRRCGMTRW-AHCXROLUSA-N Iodine-123 Chemical compound [123I] ZCYVEMRRCGMTRW-AHCXROLUSA-N 0.000 description 1
- OGNSCSPNOLGXSM-VKHMYHEASA-N L-2,4-diaminobutyric acid Chemical compound NCC[C@H](N)C(O)=O OGNSCSPNOLGXSM-VKHMYHEASA-N 0.000 description 1
- RDFMDVXONNIGBC-LURJTMIESA-N L-2-aminoheptanoic acid Chemical compound CCCCC[C@H](N)C(O)=O RDFMDVXONNIGBC-LURJTMIESA-N 0.000 description 1
- SNDPXSYFESPGGJ-BYPYZUCNSA-N L-2-aminopentanoic acid Chemical compound CCC[C@H](N)C(O)=O SNDPXSYFESPGGJ-BYPYZUCNSA-N 0.000 description 1
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 description 1
- AGPKZVBTJJNPAG-UHNVWZDZSA-N L-allo-Isoleucine Chemical compound CC[C@@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-UHNVWZDZSA-N 0.000 description 1
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 description 1
- 102100039648 Lactadherin Human genes 0.000 description 1
- 102100038609 Lactoperoxidase Human genes 0.000 description 1
- 108010023244 Lactoperoxidase Proteins 0.000 description 1
- 102000002297 Laminin Receptors Human genes 0.000 description 1
- 108010000851 Laminin Receptors Proteins 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 102100020872 Leucyl-cystinyl aminopeptidase Human genes 0.000 description 1
- 101000577064 Lymnaea stagnalis Molluscan insulin-related peptide 1 Proteins 0.000 description 1
- 102000034655 MIF Human genes 0.000 description 1
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 1
- 108010048043 Macrophage Migration-Inhibitory Factors Proteins 0.000 description 1
- 102100037791 Macrophage migration inhibitory factor Human genes 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- WAEMQWOKJMHJLA-UHFFFAOYSA-N Manganese(2+) Chemical compound [Mn+2] WAEMQWOKJMHJLA-UHFFFAOYSA-N 0.000 description 1
- 102000029749 Microtubule Human genes 0.000 description 1
- 108091022875 Microtubule Proteins 0.000 description 1
- 102100024193 Mitogen-activated protein kinase 1 Human genes 0.000 description 1
- 102000005431 Molecular Chaperones Human genes 0.000 description 1
- 108010006519 Molecular Chaperones Proteins 0.000 description 1
- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 description 1
- 101000737895 Mytilus edulis Contraction-inhibiting peptide 1 Proteins 0.000 description 1
- PQNASZJZHFPQLE-LURJTMIESA-N N(6)-methyl-L-lysine Chemical compound CNCCCC[C@H](N)C(O)=O PQNASZJZHFPQLE-LURJTMIESA-N 0.000 description 1
- OLNLSTNFRUFTLM-BYPYZUCNSA-N N-ethyl-L-asparagine Chemical compound CCN[C@H](C(O)=O)CC(N)=O OLNLSTNFRUFTLM-BYPYZUCNSA-N 0.000 description 1
- OLNLSTNFRUFTLM-UHFFFAOYSA-N N-ethylasparagine Chemical compound CCNC(C(O)=O)CC(N)=O OLNLSTNFRUFTLM-UHFFFAOYSA-N 0.000 description 1
- YPIGGYHFMKJNKV-UHFFFAOYSA-N N-ethylglycine Chemical compound CC[NH2+]CC([O-])=O YPIGGYHFMKJNKV-UHFFFAOYSA-N 0.000 description 1
- 108010065338 N-ethylglycine Proteins 0.000 description 1
- AKCRVYNORCOYQT-YFKPBYRVSA-N N-methyl-L-valine Chemical compound CN[C@@H](C(C)C)C(O)=O AKCRVYNORCOYQT-YFKPBYRVSA-N 0.000 description 1
- VEQPNABPJHWNSG-UHFFFAOYSA-N Nickel(2+) Chemical compound [Ni+2] VEQPNABPJHWNSG-UHFFFAOYSA-N 0.000 description 1
- 102000007999 Nuclear Proteins Human genes 0.000 description 1
- 108010089610 Nuclear Proteins Proteins 0.000 description 1
- 108010078627 Oncogene Protein v-crk Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 1
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 description 1
- 206010033128 Ovarian cancer Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000018546 Paxillin Human genes 0.000 description 1
- ACNHBCIZLNNLRS-UHFFFAOYSA-N Paxilline 1 Natural products N1C2=CC=CC=C2C2=C1C1(C)C3(C)CCC4OC(C(C)(O)C)C(=O)C=C4C3(O)CCC1C2 ACNHBCIZLNNLRS-UHFFFAOYSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108010081690 Pertussis Toxin Proteins 0.000 description 1
- 101710114878 Phospholipase A-2-activating protein Proteins 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 102100022427 Plasmalemma vesicle-associated protein Human genes 0.000 description 1
- 101710193105 Plasmalemma vesicle-associated protein Proteins 0.000 description 1
- 108010072866 Prostate-Specific Antigen Proteins 0.000 description 1
- 102100038358 Prostate-specific antigen Human genes 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 108700020978 Proto-Oncogene Proteins 0.000 description 1
- 102000052575 Proto-Oncogene Human genes 0.000 description 1
- 108010024221 Proto-Oncogene Proteins c-bcr Proteins 0.000 description 1
- 102000015690 Proto-Oncogene Proteins c-bcr Human genes 0.000 description 1
- 230000006819 RNA synthesis Effects 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 230000018199 S phase Effects 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 102000005588 Son of Sevenless Proteins Human genes 0.000 description 1
- 108010059447 Son of Sevenless Proteins Proteins 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 241000202349 Taxus brevifolia Species 0.000 description 1
- 229910052771 Terbium Inorganic materials 0.000 description 1
- 108700031954 Tgfb1i1/Leupaxin/TGFB1I1 Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 108020004566 Transfer RNA Proteins 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 102100031988 Tumor necrosis factor ligand superfamily member 6 Human genes 0.000 description 1
- 108050002568 Tumor necrosis factor ligand superfamily member 6 Proteins 0.000 description 1
- 102000003425 Tyrosinase Human genes 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 101710145727 Viral Fc-gamma receptor-like protein UL119 Proteins 0.000 description 1
- 108700005077 Viral Genes Proteins 0.000 description 1
- 229910052769 Ytterbium Inorganic materials 0.000 description 1
- VWQVUPCCIRVNHF-OUBTZVSYSA-N Yttrium-90 Chemical compound [90Y] VWQVUPCCIRVNHF-OUBTZVSYSA-N 0.000 description 1
- 239000003070 absorption delaying agent Substances 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 108010017893 alanyl-alanyl-alanine Proteins 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 150000008052 alkyl sulfonates Chemical class 0.000 description 1
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical class OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 150000003862 amino acid derivatives Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 229960002684 aminocaproic acid Drugs 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 239000001166 ammonium sulphate Substances 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- XCPGHVQEEXUHNC-UHFFFAOYSA-N amsacrine Chemical class COC1=CC(NS(C)(=O)=O)=CC=C1NC1=C(C=CC=C2)C2=NC2=CC=CC=C12 XCPGHVQEEXUHNC-UHFFFAOYSA-N 0.000 description 1
- 229960001220 amsacrine Drugs 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002583 anti-histone Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 229940045687 antimetabolites folic acid analogs Drugs 0.000 description 1
- 229940045719 antineoplastic alkylating agent nitrosoureas Drugs 0.000 description 1
- 239000003972 antineoplastic antibiotic Substances 0.000 description 1
- 239000003816 antisense DNA Substances 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- RYXHOMYVWAEKHL-OUBTZVSYSA-N astatine-211 Chemical compound [211At] RYXHOMYVWAEKHL-OUBTZVSYSA-N 0.000 description 1
- 238000011717 athymic nude mouse Methods 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 238000003287 bathing Methods 0.000 description 1
- 108010056708 bcr-abl Fusion Proteins Proteins 0.000 description 1
- 102000004441 bcr-abl Fusion Proteins Human genes 0.000 description 1
- 229940000635 beta-alanine Drugs 0.000 description 1
- 230000002457 bidirectional effect Effects 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000007413 biotinylation Methods 0.000 description 1
- 230000006287 biotinylation Effects 0.000 description 1
- 229910052797 bismuth Inorganic materials 0.000 description 1
- JCXGWMGPZLAOME-UHFFFAOYSA-N bismuth atom Chemical compound [Bi] JCXGWMGPZLAOME-UHFFFAOYSA-N 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 229910021538 borax Inorganic materials 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000012830 cancer therapeutic Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 150000004697 chelate complex Chemical class 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 210000003837 chick embryo Anatomy 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 229960004926 chlorobutanol Drugs 0.000 description 1
- 238000011210 chromatographic step Methods 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- BFGKITSFLPAWGI-UHFFFAOYSA-N chromium(3+) Chemical compound [Cr+3] BFGKITSFLPAWGI-UHFFFAOYSA-N 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 239000013611 chromosomal DNA Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000000749 co-immunoprecipitation Methods 0.000 description 1
- 238000011490 co-immunoprecipitation assay Methods 0.000 description 1
- 230000008045 co-localization Effects 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000009096 combination chemotherapy Methods 0.000 description 1
- 230000002301 combined effect Effects 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 238000010226 confocal imaging Methods 0.000 description 1
- 230000001268 conjugating effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000012050 conventional carrier Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- RYGMFSIKBFXOCR-AKLPVKDBSA-N copper-67 Chemical compound [67Cu] RYGMFSIKBFXOCR-AKLPVKDBSA-N 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 238000002681 cryosurgery Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 108010004073 cysteinylcysteine Proteins 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 1
- 229960003901 dacarbazine Drugs 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- YSMODUONRAFBET-UHFFFAOYSA-N delta-DL-hydroxylysine Natural products NCC(O)CCC(N)C(O)=O YSMODUONRAFBET-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- KBQHZAAAGSGFKK-UHFFFAOYSA-N dysprosium atom Chemical compound [Dy] KBQHZAAAGSGFKK-UHFFFAOYSA-N 0.000 description 1
- 230000001094 effect on targets Effects 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 230000003028 elevating effect Effects 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 108700004025 env Genes Proteins 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- UYAHIZSMUZPPFV-UHFFFAOYSA-N erbium Chemical compound [Er] UYAHIZSMUZPPFV-UHFFFAOYSA-N 0.000 description 1
- YSMODUONRAFBET-UHNVWZDZSA-N erythro-5-hydroxy-L-lysine Chemical compound NC[C@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-UHNVWZDZSA-N 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- OGPBJKLSAFTDLK-IGMARMGPSA-N europium-152 Chemical compound [152Eu] OGPBJKLSAFTDLK-IGMARMGPSA-N 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002270 exclusion chromatography Methods 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N ferric oxide Chemical compound O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 229960005191 ferric oxide Drugs 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 229960000390 fludarabine Drugs 0.000 description 1
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 150000002224 folic acids Chemical class 0.000 description 1
- 201000003444 follicular lymphoma Diseases 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 108700004026 gag Genes Proteins 0.000 description 1
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 1
- IRSCQMHQWWYFCW-UHFFFAOYSA-N ganciclovir Chemical compound O=C1NC(N)=NC2=C1N=CN2COC(CO)CO IRSCQMHQWWYFCW-UHFFFAOYSA-N 0.000 description 1
- 229960002963 ganciclovir Drugs 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 102000034356 gene-regulatory proteins Human genes 0.000 description 1
- 108091006104 gene-regulatory proteins Proteins 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- CBMIPXHVOVTTTL-UHFFFAOYSA-N gold(3+) Chemical compound [Au+3] CBMIPXHVOVTTTL-UHFFFAOYSA-N 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000003505 heat denaturation Methods 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 238000001239 high-resolution electron microscopy Methods 0.000 description 1
- SCKNFLZJSOHWIV-UHFFFAOYSA-N holmium(3+) Chemical compound [Ho+3] SCKNFLZJSOHWIV-UHFFFAOYSA-N 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- QJHBJHUKURJDLG-UHFFFAOYSA-N hydroxy-L-lysine Natural products NCCCCC(NO)C(O)=O QJHBJHUKURJDLG-UHFFFAOYSA-N 0.000 description 1
- 229960001330 hydroxycarbamide Drugs 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 238000012872 hydroxylapatite chromatography Methods 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 229960000908 idarubicin Drugs 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 229940127121 immunoconjugate Drugs 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 230000002621 immunoprecipitating effect Effects 0.000 description 1
- 229910052738 indium Inorganic materials 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 239000003701 inert diluent Substances 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 108060004057 integrin beta chain Proteins 0.000 description 1
- 102000017776 integrin beta chain Human genes 0.000 description 1
- 230000035990 intercellular signaling Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- XMBWDFGMSWQBCA-YPZZEJLDSA-N iodane Chemical compound [125IH] XMBWDFGMSWQBCA-YPZZEJLDSA-N 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N iron oxide Inorganic materials [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 1
- 235000013980 iron oxide Nutrition 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002540 isothiocyanates Chemical class 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 229940057428 lactoperoxidase Drugs 0.000 description 1
- CZMAIROVPAYCMU-UHFFFAOYSA-N lanthanum(3+) Chemical compound [La+3] CZMAIROVPAYCMU-UHFFFAOYSA-N 0.000 description 1
- 238000002430 laser surgery Methods 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 description 1
- 229960004844 lovastatin Drugs 0.000 description 1
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000010946 mechanistic model Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000002082 metal nanoparticle Substances 0.000 description 1
- 239000002923 metal particle Substances 0.000 description 1
- 150000001455 metallic ions Chemical class 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000010232 migration assay Methods 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 210000001700 mitochondrial membrane Anatomy 0.000 description 1
- 229960001156 mitoxantrone Drugs 0.000 description 1
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical group O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004001 molecular interaction Effects 0.000 description 1
- 238000000302 molecular modelling Methods 0.000 description 1
- 125000004573 morpholin-4-yl group Chemical group N1(CCOCC1)* 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 229940087004 mustargen Drugs 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- QEFYFXOXNSNQGX-UHFFFAOYSA-N neodymium atom Chemical compound [Nd] QEFYFXOXNSNQGX-UHFFFAOYSA-N 0.000 description 1
- 210000005170 neoplastic cell Anatomy 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- OSTGTTZJOCZWJG-UHFFFAOYSA-N nitrosourea Chemical compound NC(=O)N=NO OSTGTTZJOCZWJG-UHFFFAOYSA-N 0.000 description 1
- 108091027963 non-coding RNA Proteins 0.000 description 1
- 102000042567 non-coding RNA Human genes 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 231100000590 oncogenic Toxicity 0.000 description 1
- 230000002246 oncogenic effect Effects 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229960003104 ornithine Drugs 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000011499 palliative surgery Methods 0.000 description 1
- 230000005298 paramagnetic effect Effects 0.000 description 1
- ACNHBCIZLNNLRS-UBGQALKQSA-N paxilline Chemical compound N1C2=CC=CC=C2C2=C1[C@]1(C)[C@@]3(C)CC[C@@H]4O[C@H](C(C)(O)C)C(=O)C=C4[C@]3(O)CC[C@H]1C2 ACNHBCIZLNNLRS-UBGQALKQSA-N 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000008823 permeabilization Effects 0.000 description 1
- 102000013415 peroxidase activity proteins Human genes 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 238000005191 phase separation Methods 0.000 description 1
- 229960003742 phenol Drugs 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- BLFWHYXWBKKRHI-JYBILGDPSA-N plap Chemical compound N([C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(=O)[C@@H]1CCCN1C(=O)[C@H](CO)NC(=O)[C@@H](N)CCC(O)=O BLFWHYXWBKKRHI-JYBILGDPSA-N 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 108700004029 pol Genes Proteins 0.000 description 1
- DTBMTXYWRJNBGK-UHFFFAOYSA-L potassium;sodium;phthalate Chemical compound [Na+].[K+].[O-]C(=O)C1=CC=CC=C1C([O-])=O DTBMTXYWRJNBGK-UHFFFAOYSA-L 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 125000001500 prolyl group Chemical group [H]N1C([H])(C(=O)[*])C([H])([H])C([H])([H])C1([H])[H] 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 238000002731 protein assay Methods 0.000 description 1
- 239000003528 protein farnesyltransferase inhibitor Substances 0.000 description 1
- 238000001273 protein sequence alignment Methods 0.000 description 1
- 238000000734 protein sequencing Methods 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000010837 receptor-mediated endocytosis Effects 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012508 resin bead Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000003938 response to stress Effects 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- WUAPFZMCVAUBPE-IGMARMGPSA-N rhenium-186 Chemical compound [186Re] WUAPFZMCVAUBPE-IGMARMGPSA-N 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- DOSGOCSVHPUUIA-UHFFFAOYSA-N samarium(3+) Chemical compound [Sm+3] DOSGOCSVHPUUIA-UHFFFAOYSA-N 0.000 description 1
- 229940043230 sarcosine Drugs 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 125000005630 sialyl group Chemical group 0.000 description 1
- 230000007781 signaling event Effects 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 235000009518 sodium iodide Nutrition 0.000 description 1
- 235000010339 sodium tetraborate Nutrition 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000010473 stable expression Effects 0.000 description 1
- 238000011301 standard therapy Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000011477 surgical intervention Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 101150047061 tag-72 gene Proteins 0.000 description 1
- 229910052713 technetium Inorganic materials 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- NRUKOCRGYNPUPR-QBPJDGROSA-N teniposide 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](OC[C@H]4O3)C=3SC=CC=3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 NRUKOCRGYNPUPR-QBPJDGROSA-N 0.000 description 1
- 229960001278 teniposide Drugs 0.000 description 1
- GZCRRIHWUXGPOV-UHFFFAOYSA-N terbium atom Chemical compound [Tb] GZCRRIHWUXGPOV-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- YSMODUONRAFBET-WHFBIAKZSA-N threo-5-hydroxy-L-lysine Chemical compound NC[C@@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-WHFBIAKZSA-N 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 230000036964 tight binding Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 210000003437 trachea Anatomy 0.000 description 1
- BJBUEDPLEOHJGE-IMJSIDKUSA-N trans-3-hydroxy-L-proline Chemical compound O[C@H]1CC[NH2+][C@@H]1C([O-])=O BJBUEDPLEOHJGE-IMJSIDKUSA-N 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- 230000007723 transport mechanism Effects 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- 150000003918 triazines Chemical class 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- BSVBQGMMJUBVOD-UHFFFAOYSA-N trisodium borate Chemical compound [Na+].[Na+].[Na+].[O-]B([O-])[O-] BSVBQGMMJUBVOD-UHFFFAOYSA-N 0.000 description 1
- 231100000588 tumorigenic Toxicity 0.000 description 1
- 230000000381 tumorigenic effect Effects 0.000 description 1
- 238000013042 tunel staining Methods 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 239000002691 unilamellar liposome Substances 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 229910052720 vanadium Inorganic materials 0.000 description 1
- LEONUFNNVUYDNQ-UHFFFAOYSA-N vanadium atom Chemical compound [V] LEONUFNNVUYDNQ-UHFFFAOYSA-N 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 230000007998 vessel formation Effects 0.000 description 1
- JXLYSJRDGCGARV-CFWMRBGOSA-N vinblastine 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-CFWMRBGOSA-N 0.000 description 1
- GBABOYUKABKIAF-IELIFDKJSA-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-IELIFDKJSA-N 0.000 description 1
- 229960002066 vinorelbine Drugs 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- NAWDYIZEMPQZHO-UHFFFAOYSA-N ytterbium Chemical compound [Yb] NAWDYIZEMPQZHO-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70546—Integrin superfamily
- C07K14/7055—Integrin beta1-subunit-containing molecules, e.g. CD29, CD49
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
Definitions
- the present invention concerns the fields of molecular medicine and targeted delivery of therapeutic and detecting agents. More specifically, the present invention relates to the identification of novel peptide sequences that selectively target cancers for the treatment and detection of cancer.
- Cancer therapeutic agents in particular exhibit a very low therapeutic index, with rapidly growing normal tissues such as skin and bone marrow typically affected at concentrations of agent that are not much higher than the concentrations used to kill tumor cells.
- Treatment and diagonosis of cancer would be greatly facilitated by the development of compositions and methods for targeted delivery to cancer cells, more specifically, by using antibodies or tumor-homing peptides that bind to targets found on the surfaces of cancer cells, but not found on normal tissues. Such targets, which must have minimal homology to other cell surface molecules, are difficult to find.
- phage display libraries to identify organ or tissue targeting peptides in a mouse model system.
- Such libraries can be generated by inserting random oligonucleotides into cDNAs encoding a phage surface protein, generating collections of phage particles displaying unique peptides in as many as 10e9 permutations (Pasqualini and Ruoslahti, 1996, Arap et al., 1998; Pasqualini et al., 2001).
- Intravenous administration of phage display libraries to tumor bearing mice was followed by the recovery of phage from tumor xenograft and the tumor targetting peptides capable of selective homing to the tumor were characterzied.
- Phage were recovered that were capable of selective homing to the vascular beds of different mouse organs or tissues, based on the specific targeting peptide sequences expressed on the outer surface of the phage (Pasqualini and Ruoslahti, 1996). Each of those tumor-homing peptides bound to receptors that were selectively expressed or upregulated in the surface of tumor cells.
- Attachment of therapeutic agents to targeting peptides resulted in the selective delivery of the agent to a desired organ or tissue in the mouse model system.
- Targeted delivery of chemotherapeutic agents and proapoptotic peptides to receptors located in tumor angiogenic vasculature resulted in a marked increase in therapeutic efficacy and a decrease in systemic toxicity in tumor-bearing mouse models (Arap et al., 1998a, 1998b; Ellerby et al., 1999).
- CRKL (chicken tumor virus number 10 regulator of kinase-like protein), an adapter protein, is a homolog of oncogene v-crk. It contains one SH2 domain and two tandem SH3 domains. Intracellular CRKL is implicated in both MAP kinase and integrin- mediated pathways (Li et al., 2003; Uemura et al., 1999). In addition, CRKL has an oncogenic potential.
- the present invention overcomes deficiencies in the prior art by providing methods and compositions for selectively targeting secreted CRKL through the use of targeting peptides.
- Selective targeting of secreted CRKL through the use of a targeting peptide may be used, for example, in the treatment of cancer to deliver a chemotherapeutic compound, fusion protein, or fusion construct to a cancer cell or tissue.
- a targeting peptide may be used, for example, in the treatment of cancer to deliver a chemotherapeutic compound, fusion protein, or fusion construct to a cancer cell or tissue.
- CRKL targets the plexin-semaphorin-integrin (PSI) domain of ⁇ i integrin located outside of the cell, triggers MAP kinases, and promotes cell growth and survival.
- PSI plexin-semaphorin-integrin
- An aspect of the present invention related to an isolated tumor targeting peptide comprising a CRKL binding motif, said motif defined as: being from 6 to 20 amino acids in length, having a degree of similarity to a corresponding best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47) of at least 25%; and wherein the targeting peptide is 100 amino acids or less in length and binds under physiological conditions to cells expressing CRKL.
- the CRKL binding motif may have a degree of similarity to a best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47) of at least 40%, at least 50%, or at least 60%.
- the peptide has a sequence that is not identical to a best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47).
- the CRKL binding motif may have a best fit sequence alignment to a ⁇ l integrin (SEQ ID NO:47) PSI domain region.
- the CRKL binding motif may have a best fit sequence alignment to a ⁇ l integrin (SEQ ID NO:47) PSI domain region selected from the group consisting of amino acids 6 to 10, 10 to 29; 15 to 34; 18 to 37; 36 to 55; 39 to 58; 45 to 64; 94 to 113; 196 to 215; 198 to 213; 203 to 222; 244 to 263; 330 to 349; 377 to 396; 379 to 398; 380 to 399; 398 to 417; 400 to 419; 413 to 432; 447 to 466; 460 to 479; 460 to 479; 464 to 483; 469 to 488; 474 to 493; 475 to 494; 512 to 533; 519 to 538; 551 to 570; 574 to 593; 577 to 596; 579 to 598; 590 to 609; 596 to 615; 613 to 632; 615 to 634; 616 to 635; 644
- the isolated peptide may be further defined as a cyclic peptide that are capable of being prepared in a cyclic form, such as peptide having a cysteine residue ("C") at both termini, which may, where desired, be provided in cyclic form, such as through the formation of a di-cysteine (i.e., cystine).
- C cysteine residue
- Such cyclic peptides may be of particular importance in that disulfide bonds in peptides makes them remarkably stable to chemical, thermal or enzymatic degradation.
- Such cyclic peptides may be of particular importance in therapeutic and diagnostic applications, where poor availability, susceptibility to proteolysis and short in vivo half- lives are concerned.
- Said peptide may be attached to a molecule; for example, the molecule may be a protein and the peptide may be conjugated or fused to the protein to form a protein conjugate, wherein the protein conjugate is not a naturally occurring protein.
- the peptide may be positioned at a terminus of the protein.
- Said molecule may be a pro-apoptosis agent, an anti-angiogenic agent, a cytokine, a cytotoxic agent, a drug, a chemotherapeutic agent, a hormone, a growth factor, an antibiotic, an antibody or fragment or single chain thereof, a survival factor, an anti-apoptotic agent, a hormone antagonist, an antigen, a peptide, a protein, a diagnostic agent, a radioisotope, or an imaging agent.
- Said molecule may be a pro- apoptosis agent selected from the group consisting of gramicidin; magainin; mellitin; defensin; cecropin; (KLAKLAK) 2 (SEQ ID NO:48); (KLAKKLA) 2 (SEQ ID NO:49); (KAAKKAA) 2 (SEQ ID NO:50); (KLGKKLG) 3 (SEQ ID NO:51); Bcl-2; Bad; Bak; Bax; and Bik.
- said pro-apoptosis agent is (KLAKLAK) 2 (SEQ ID NO:48).
- SEQ ID NO:48 may consist of D amino acids.
- said molecule may be an anti-angiogenic agent selected from the group consisting of thrombospondin, an angiostatin, pigment epithelium-derived factor, angiotensin, laminin peptides, fibronectin peptides, plasminogen activator inhibitors, tissue metalloproteinase inhibitors, interferons, interleukin 12, platelet factor 4, IP-10, Gro- ⁇ , thrombospondin, 2-methoxyoestradiol, proliferin-related protein, carboxiamidotriazole,
- CMlOl, Marimastat pentosan polysulphate, angiopoietin 2, herbimycin A, PNU145156E, 16K prolactin fragment, Linomide, thalidomide, pentoxifylline, genistein, TNP-470, endostatin, paclitaxel, Docetaxel, polyamines, a proteasome inhibitor, a kinase inhibitor, a signaling peptide, accutin, cidofovir, vincristine, bleomycin, AGM- 1470, platelet factor 4, minocycline, endostatin XVIII, endostatin XV, the C-terminal hemopexin domain of matrix metalloproteinase-2, the kringle 5 domain of human plasminogen, a fusion protein of endostatin and angiostatin, a fusion protein of endostatin and the kringle 5 domain of human plasminogen, the monokine-induced
- Said molecule may be a cytokine selected from the group consisting of interleukin 1 (IL-I), IL-2, IL-5, IL-IO, IL-11, IL-12, IL-18, interferon- ⁇ (IF- ⁇ ), IF- ⁇ , IF- ⁇ , a tumor necrosis factor, or GM-CSF (granulocyte macrophage colony stimulating factor).
- IL-I interleukin 1
- IL-2 interleukin 2
- IL-5 IL-IO
- IL-11 IL-12
- IL-18 interferon- ⁇
- IF- ⁇ interferon- ⁇
- IF- ⁇ IF- ⁇
- IF- ⁇ tumor necrosis factor
- GM-CSF granulocyte macrophage colony stimulating factor
- Said peptide may be attached to a macromolecular complex, such as a virus, a bacteriophage, a bacterium, a liposome, a microparticle, a magnetic bead, a yeast cell, or a mammalian cell.
- a virus such as a lentivirus, papovavirus, adenovirus, retrovirus, AAV, vaccinia virus or herpes virus.
- Said peptide may be attached to a solid support, such as a microtiter dish or microchip.
- Another aspect of the present invention relates to a method of preparing a construct comprising obtaining a peptide in accordance with the present invention and attaching the peptide to a molecule to prepare the construct.
- Yet another aspect of the invention relates to a method of targeting the delivery of a peptide, molecule or protein to cells that express CRKL, the method comprising the steps of: obtaining a peptide according to the present invention, or prepared by the above method, and administering the peptide to a cell population, wherein the population includes cells that express CRKL, to thereby deliver the molecule or protein to said cells.
- the cells that express CRKL may be in a subject, and the peptide or protein fusion construct may be formulated in a pharmaceutically acceptable composition and the composition may be administered to the subject.
- the subject may be a human subject.
- the method is further defined as a detection method and the method further comprises detecting the peptide, molecule or protein that has been delivered to the cells.
- the subject may have a disease or disorder and the method may be further defined as a therapeutic method.
- the subject may have a cancer, such as a cancer of the prostate, breast, sarcoma, gum, tongue, lung, skin, liver, kidney, eye, brain, leukemia, mesothelioma, neuroblastoma, head, neck, pancreatic, renal, bone, testicular, ovarian, mesothelioma, cervical, gastrointestinal, lymphoma, brain, colon and bladder.
- Embodiments discussed in the context of a methods and/or composition of the invention may be employed with respect to any other method or composition described herein. Thus, an embodiment pertaining to one method or composition may be applied to other methods and compositions of the invention as well.
- FIG. IA-C Peptide targeting and internalizarion in cancer cells.
- FIG. IA the phage peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) binds to the cell surface on DU145 cells.
- FIG. IB immunolocalization of tumor-homing phage on the cell surface of non-permeabilized KS 1767 cells.
- Synthetic peptide YRCTLNSPFFWEDMTHECHAGG SEQ ID NO:67
- D KLAKLAK 2
- FIG. 2A-C Sequence alignment of tumor-homing peptides and P 1 integrin (SEQ ID NO:13).
- FIG. 2A the tumor-homing peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) matches to the plexin-semaphorin-integrin (PSI) domain (sequence region 26-78 residues) (SEQ ID NO: 12).
- FIG. 2B sequence alignment of all eight ⁇ integrin- subunits (SEQ ID NO: 14-21) and the YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) peptide sequence.
- FIG. 2C sequence alignment of all the tumor homing peptides (Table 1) and ⁇ i integrin (SEQ ID NO: 13).
- FIG. 3A-D Receptor binding by peptides.
- FIG. 3A the recombinant His-tag CRKL (rCRKL), rCRKL-SH3 (N) domain, and rCRKL-SH3 (C) domain bind to the tumor- homing peptide-YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1).
- FIG. 3B the recombinant His-tag rCRKL, rCRKL-SH3 (N) domain, and rCRKL-SH3 (C) domain bind to the synthetic peptide NSTFLQEGMPTSA (SEQ ID NO:23) corresponding to a region in the PSI domain.
- FIG. 3A the recombinant His-tag CRKL (rCRKL), rCRKL-SH3 (N) domain, and rCRKL-SH3 (C) domain bind to the synthetic peptide NSTFLQEGMPTSA (SEQ ID NO:23
- the binding activity of the tumor-homing peptide to rCRKL-SH3 (C) domain is inhibited by the tumor- homing (YRCTLNSPFFWEDMTHECHA; SEQ ID NO:1) peptide, by the PSI-derived (NSTFLQEGMPTSA; SEQ ID NO:23) peptide or tumor-homing phage displaying SEQ ID NO: 1. Bars represent mean ⁇ standard deviation from triplicate wells.
- FIG. 4A-D The interaction between CRKL and binding peptides.
- FIG. 4 A the binding properties of the rCRKL-SH3 (C) to the tumor-homing peptide.
- a Pro— »Ala in SEQ ID NO:1 is provided as SEQ ID NO:25. Mutational analysis of the CRKL SH3 (C) domain.
- FIG. 4B tumor-homing phage (displaying YRCTLNSPFFWEDMTHECHA; SEQ ID NO:25.
- FIG. 4C scheme of the CRKL SH3 (C) domain (SEQ ID NO:24) and the control deletion mutants generated as His-tag recombinant proteins. Four mutants were generated and tested: ⁇ l (deleted residues 236- 256), ⁇ 2 (deleted residues 257-277), ⁇ 3 (deleted residues 278- 293), and ⁇ SH3 (C) (deleted residues 236-293).
- FIG. 4D the binding region is located between residues 236-277 of the SH3 (C) domain.
- FIG. 5 Protein-protein interaction between CRKL and ⁇ l integrin. A concentration-dependent inhibition of CRKL binding to ⁇ i integrin by recombinant gst-PSI protein (up to 800 ⁇ m). The integrins ⁇ v ⁇ 3 and ⁇ v ⁇ 5 served as controls. Standard deviations of the mean from triplicate wells are shown.
- FIG. 6A-B Cell surface localization of CRKL.
- FIG. 6A flow cytometry analysis of CRKL on DU145 cells. Immunolabeling was performed by using monoclonal anti-CRKL (**), anti- ⁇ i integrin (*), and anti-AHSG antibodies (c, control).
- FIG. 6B transmission electron microscopy (TEM) of CRKL showing individual CRKL-gold particles on the cell surface (arrow heads).
- DU 145 cells used in studies were fixed without permeabilization.
- An anti-CRKL polyclonal antibody was used in studies. Scale bars are indicated.
- FIG. 7A-C CRKL secretion.
- FIG. 7A Various cancer cell types cultured in serum free media (SFM) secrete the unphosphorylated form of CRKL.
- FIG. B,C CRKL antibody neutralizes the extracellular form of CRKL in the medium and affects cell proliferation and migration.
- Control antibodies used were as follows: anti-ILl l receptor, anti-AHSG, anti-grb2, anti- ⁇ , 6 integrin, and pre -immune antibodies. Bars represent mean ⁇ standard deviation from duplicate wells.
- FIG. 8A-D The effects of siRNA knockdown of CRKL.
- Cell proliferation FIG. 8A
- adhesion FIG. 8B
- migration FIG. 8C
- Standard deviations of the mean from triplicate wells are shown.
- FIG. 8D recombinant CRKL rescues CRKL siRNA knockdown cells in cell proliferation assays.
- DU145 cells were transfected with CRKL siRNA for 48 hours prior to exogenously adding recombinant CRKL to the wells. Cell proliferation was determined with the WST-I reagent.
- FIG. 9A-D Tumor targeting and mechanistic model.
- FIG. 9A-D Tumor targeting and mechanistic model.
- FIG. 9A in vitro phage binding of tumor-homing or controls (insertless, mutant (YRCTLNSAFFWEDMTHECHA; SEQ ID NO:25), or scrambled (#1 : YRFCTSPFHEWHLENTDMCA; SEQ ID NO:26, #2: YRECTDSPHEFHLWNTMCAF; SEQ ID NO:27)) phage on rCRKL.
- FIG. 9B-D in vivo homing of targeted or control phage constructs in mice bearing different tumor types. Tumor-homing phage localized to tumors preferentially when compared to controls. Representative data from two independent experiments are shown.
- FIG. 10 Targeting inhibition in tumor-bearing mice.
- Tumor-homing phage was pre -incubated with control gst or recombinant gst-CRKL prior to administering to nude mice bearing size matched human tumors (DU 145 -derived). Inhibition was observed in the pre-treated tumor-homing phage by recombinant CRKL. Results from two independent experiments are shown.
- FIG. 11 Treatment of tumor xenografts by a synthetic tumor-homing proapoptotic peptide. Cohorts of size-matched nude mice bearing human prostate cancer xenografts (DU 145 -derived) were used. Markedly reduced tumor growth was observed in tumor-bearing mice treated with synthetic tumor-homing proapoptotic peptide
- YRCTLNSPFFWEDMTHECHAGG SEQ ID NO:67
- D KLAKLAK 2
- Equimolar amounts of YRCTLNSPFFWEDMTHECHA (SEQ ID NO: 1) or D (KLAKLAK) 2 showed no differences in tumor volume compared to untreated animals (Student's t-test, p ⁇ 0.001).
- the present invention provides both data supporting the importance and function of intracellular signaling protein CRKL as well as peptides which target CRKL.
- Cell membranes have evolved to interface a tight and compartmentalized control between the intracellular contents and the extracellular milieu (Conner and Schmid, 2003; Cho and Stahelin, 2005).
- transmembrane receptor families mediate bidirectional signaling across the cell surface through a complex spatial and temporal organization of transduction cascades (Martin et ah, 2002; Manning et ah, 2002).
- location of the proteins involved in signal transduction is central to provide specificity of the cellular responses (Cho and Stahelin, 2005; Mochly-Rosen, 1995).
- cell surface receptors such as integrins undergo conformational changes elicited through ligand-binding to enable a cross-talk with signal transduction cascades such as mitogen-activated protein (MAP) kinase pathways; conventional integrin ligands include extracellular matrix (ECM) proteins to their extracellular domains and cytoskeletal proteins to their intracellular domains (Martin et al, 2002; Manning et al, 2002; Hunter, 2000; Pawson and Scott, 1997; Blume- Jensen and Hunter, 2001).
- ECM extracellular matrix
- CRKL targets the plexin- semaphorin-integrin (PSI) domain of ⁇ l integrin located outside of the cell, triggers MAP kinases, and promotes cell growth and survival.
- PSI plexin- semaphorin-integrin
- the below data demonstrates that intracellular unphosphorylated CRKL is either secreted by a non-classic active transport (perhaps via ABC-transporters) and/or released through cell death into the tumor microenvironment (step 1), where its SH3 domains specifically bind to the PSI domain of the ⁇ l integrin on the tumor cell surface (step 2).
- the ⁇ l integrin conformation is changed from bent to extended (active), thus triggering downstream phosphorylation of target proteins in the integrin-mediated pathway (steps 3 and 4) and/or MAP kinase pathway (steps 5-7) and ultimately affecting tumor cell migration and proliferation (step 8).
- the present invention provides isolated tumor targeting peptides comprising a CRKL binding motif, and said motif defined as being, e.g. , from 6 to 20 amino acids in length, having a degree of similarity to a corresponding best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47) of, e.g., at least 25%; and wherein the targeting peptide may be 100 amino acids or less in length and binds under physiological conditions to cells expressing CRKL.
- the CRKL binding motif may have a degree of similarity to a best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47) of at least 40%, at least 50%, or at least 60%.
- the peptide has a sequence that is not identical to a best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47).
- the CRKL binding motif may have a best fit sequence alignment to a ⁇ l integrin (SEQ ID NO:47) PSI domain region.
- the CRKL binding motif may have a best fit sequence alignment to a ⁇ l integrin (SEQ ID NO:47) PSI domain region selected from the group consisting of amino acids 10 to 29; 15 to 34; 18 to 37; 36 to 55; 39 to 58; 45 to 64; 94 to 113; 196 to 215; 198 to 213; 203 to 222; 244 to 263; 330 to 349; 377 to 396; 379 to 398; 380 to 399; 398 to 417; 400 to 419; 413 to 432; 447 to 466; 460 to 479; 460 to 479; 464 to 483; 469 to 488; 474 to 493; 475 to 494; 512 to 533; 519 to 538; 551 to 570; 574 to 593; 577 to 596; 579 to 598; 590 to 609; 596 to 615; 613 to 632; 615 to 634; 616 to 635; 644 to 663
- CRKL binding peptides may be used with the present invention.
- a non- limiting list of CRKL binding peptides is provided below (SEQ ID: 1-46).
- a "targeting moiety” is a term that encompasses various types of affinity reagents that may be used to enhance the localization or binding of a substance to a particular location in an animal, including organs, tissues, particular cell types, diseased tissues or tumors. Tareting moieties may include peptides, peptide mimetics, polypeptides, antibodies, antibody-like molecules, nucleic acids, aptamers, and fragments thereof. In certain embodiments, a targeting moiety will enhance the localization of a substance to cells expressing CRKL extracellularly, i.e., CRKL being associated with the cell surface or associated with surrounding extracelluar matrix.
- a targeting moiety of the present invention e.g., a targeting peptide
- a targeting moiety is still considered to selectively bind even if it also binds to other proteins that are not substantially homologous with the target so long as such proteins share homology with a fragment or domain of the peptide target of the antibody.
- target moiety binding to the target is still selective despite some degree of cross-reactivity. Typically, the degree of cross-reactivity can be determined and differentiated from binding to the target.
- a "tumor targeting peptide” is a peptide comprising comprising a CRKL binding motif, said motif defined as: being from 6 to 20 amino acids in length, having a degree of similarity to a corresponding best fit sequence alignment to ⁇ l integrin (SEQ ID NO:47) of at least 25%; and wherein the targeting peptide is 100 amino acids or less in length and characterized by selective localization to an organ, tissue or cell type under physiological conditions, which includes specific binding with an extracellar CRKL. Selective localization may be determined, for example, by methods disclosed below, wherein the putative targeting peptide sequence binds to a protein that is displayed on the outer surface of a phage.
- a "subject” refers generally to a mammal. In certain preferred embodiments, the subject is a mouse or rabbit. In even more preferred embodiments, the subject is a human.
- CRKL thick tumor virus number 10 regulator of kinase-like protein
- Chronic myelogenous leukemia is a hematologic malignancy in which uncontrolled proliferation of granulocytes occurs. It often is characterized by the reciprocal translocation of chromosomes 9 and 22, which relocates the Ableson (abl) protooncogene onto the 3'-end of the breakpoint cluster region (bcr). This produces a chimeric bcr-abl gene encoding a p210. sup.
- bcr-abl fusion protein which is tumorigenic and is necessary for the growth of CML cells (Szczylik et a/., 1991; Skorski et al, 1994; Tari et al, 1994; McGahon et al, 1994; Bedi et al, 1994).
- the bcr-abl protein can autophosphorylate at the 177 tyrosine amino acid found within the first exon of bcr.
- the bcr domain of the bcr-abl protein binds to the SH2 domain of the growth factor receptor-bound protein 2 (Grb2) adaptor protein.
- Grb2 binds to the human Son of sevenless 1 (hSosl) GDP/GTP exchange factor resulting in ras protein activation.
- the bcr-abl protein can also transphorylate the 177 tyrosine amino acid found within the normal bcr protein.
- Crk-like (CRKL) another adaptor protein, also has been found to bind to bcr- abl. Unlike Grb2, CRKL binds to bcr-abl through the abl domain. Through its SH3 domain, CRKL can also bind to hSosl, which again leads to Ras protein activation (ten Hoeve et al. , 1994a and 1994b).
- the bcr-abl protein has been linked to ras activation, which is known to lead to tumorigenesis.
- ras protein expression is inhibited, proliferation of CML cells is also inhibited. Therefore, one of the major pathways in which bcr-abl protein promotes CML proliferation is by activating ras protein (Skorski et al., 1994 and 1995).
- CRKL intracellular signaling protein
- a regulatory (rather than ligand-binding) ⁇ i integrin extracellular domain was observed.
- CRKL conventionally known as intracellular adaptors, targets the plexin-semaphorin-integrin (PSI) domain of ⁇ i integrin located outside of the cell, triggers MAP kinases, and promotes cell growth and survival.
- PSI plexin-semaphorin-integrin
- Sequence similarity is defined as sequence identity between two nucleotide sequences, but does not necessarily indicate that two sequences have common ancestry. For example, 25% similarity means that 25 nucleotide positions out of 100 are identical in the two nucleotide sequences.
- "Best fit sequence alignment" in the present invention is defined as a sequence analysis by using sequence alignment methods known to those of ordinary skill in the art to find the best-matching piecewise (local) or global alignments of two or more sequences. As would be known to one of skill, various algorithms may be used for sequence comparison, e.g., BLAST alignments, etc.
- Peptide Match software codified in Perl 5.8.1 based on RELIC50.
- the program calculates similarity based on a predefined residue window size between an affinity selected peptide sequence and the target protein sequence from N- to C- protein terminus in one-residue shifts.
- the peptide-protein similarity scores for each residue were calculated based on a BLOSUM62 amino acid substitution matrix modified to adjust for rare amino acid representation. Thresholds may be set at least 4 identical residues between the peptide and the protein segment to discriminate significant similarities from nonspecific background matches.
- the present invention concerns novel compositions comprising at least one protein or peptide.
- a protein or peptide generally refers, but is not limited to, a protein of greater than about 200 amino acids, up to a full length sequence translated from a gene; a polypeptide of greater than about 100 amino acids; and/or a peptide of from about 3 to about 100 amino acids.
- proteins proteins
- polypeptide and “peptide” are used interchangeably herein.
- the size of at least one protein or peptide may comprise, but is not limited to, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about
- the size of a tumor targeting peptide defined in the present invention may comprise, but is not limited to, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 amino acid residues. In other aspects the size of a tumor targeting peptide may comprise, but is not limited to, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 amino acid residues, or any range derivable therein. In certain embodiments, peptides less than or equal to 20 amino acids, or peptides 6-10 amino acids in length may be used.
- amino acid residue refers to any naturally occurring amino acid, any amino acid derivative or any amino acid mimic known in the art.
- residues of the protein or peptide are sequential, without any non-amino acid interrupting the sequence of amino acid residues.
- sequence may comprise one or more non-amino acid moiety.
- sequence of residues of the protein or peptide may be interrupted by one or more non-amino acid moieties.
- protein or peptide encompasses amino acid sequences comprising at least one of the 20 common amino acids found in naturally occurring proteins, or at least one modified or unusual amino acid, including but not limited to Aad, 2-Aminoadipic acid; EtAsn, N-Ethylasparagine; Baad, 3- Aminoadipic acid, HyI, Hydroxylysine; BaIa, ⁇ -alanine, ⁇ -Amino-propionic acid; AHyI, allo-Hydroxylysine; Abu, 2-Aminobutyric acid; 3Hyp, 3-Hydroxyproline; 4Abu, 4- Aminobutyric acid, piperidinic acid; 4Hyp, 4-Hydroxyproline; Acp, 6-Aminocaproic acid, Ide, Isodesmosine; Ahe, 2-Aminoheptanoic acid; AIIe, allo-Isoleucine; Aib
- Proteins or peptides may be made by any technique known to those of skill in the art, including the expression of proteins, polypeptides or peptides through standard molecular biological techniques, the isolation of proteins or peptides from natural sources, or the chemical synthesis of proteins or peptides.
- the nucleotide and protein, polypeptide and peptide sequences corresponding to various genes have been previously disclosed, and may be found at computerized databases known to those of ordinary skill in the art.
- One such database is the National Center for Biotechnology Information's Genbank and GenPept databases (world wide web at ncbi.nlm.nih.gov).
- the coding regions for known genes may be amplified and/or expressed using the techniques disclosed herein or as would be know to those of ordinary skill in the art. Alternatively, various commercial preparations of proteins, polypeptides and peptides are known to those of skill in the art.
- fusion proteins generally have all or a substantial portion of a tumor targeting peptide, linked at the N- or C-terminus, to all or a portion of a second polypeptide or protein.
- fusions may employ leader sequences from other species to permit the recombinant expression of a protein in a heterologous host.
- Another useful fusion includes the addition of an immunologically active domain, such as an antibody epitope, to, for example, facilitate purification of the fusion protein. Inclusion of a cleavage site at or near the fusion junction will facilitate removal of the extraneous polypeptide after purification.
- fusion proteins include linking of functional domains, such as active sites from enzymes, glycosylation domains, cellular targeting signals or transmembrane regions.
- the fusion proteins of the instant invention comprise an LPR targeting peptide linked to a therapeutic protein or peptide.
- proteins or peptides that may be incorporated into a fusion protein include cytostatic proteins, cytocidal proteins, pro-apoptosis agents, anti- angiogenic agents, hormones, cytokines, growth factors, peptide drugs, antibodies, Fab fragments antibodies, antigens, receptor proteins, enzymes, lectins, MHC proteins, cell adhesion proteins and binding proteins.
- fusion protein comprising a targeting peptide.
- Methods of generating fusion proteins are well known to those of skill in the art. Such proteins can be produced, for example, by chemical attachment using bifunctional cross-linking reagents, by de novo synthesis of the complete fusion protein, or by attachment of a DNA sequence encoding the targeting peptide to a DNA sequence encoding the second peptide or protein, followed by expression of the intact fusion protein.
- a protein or peptide may be isolated or purified.
- Protein purification techniques are well known to those of skill in the art. These techniques involve, at one level, the homogenization and crude fractionation of the cells, tissue or organ to polypeptide and non-polypeptide fractions.
- the protein or polypeptide of interest may be further purified using chromatographic and electrophoretic techniques to achieve partial or complete purification (or purification to homogeneity).
- Analytical methods particularly suited to the preparation of a pure peptide are ion-exchange chromatography, gel exclusion chromatography, polyacrylamide gel electrophoresis, affinity chromatography, immunoaffinity chromatography and isoelectric focusing.
- receptor protein purification by affinity chromatography is disclosed in U.S. Patent 5,206,347, the entire text of which is incorporated herein by reference.
- a particularly efficient method of purifying peptides is fast performance liquid chromatography (FPLC) or even high performance liquid chromatography (HPLC).
- a purified protein or peptide is intended to refer to a composition, isolatable from other components, wherein the protein or peptide is purified to any degree relative to its naturally-obtainable state.
- An isolated or purified protein or peptide therefore, also refers to a protein or peptide free from the environment in which it may naturally occur.
- purified will refer to a protein or peptide composition that has been subjected to fractionation to remove various other components, and which composition substantially retains its expressed biological activity.
- substantially purified this designation will refer to a composition in which the protein or peptide forms the major component of the composition, such as constituting about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or more of the proteins in the composition.
- Various methods for quantifying the degree of purification of the protein or peptide are known to those of skill in the art in light of the present disclosure. These include, for example, determining the specific activity of an active fraction, or assessing the amount of polypeptides within a fraction by SDS/PAGE analysis.
- a preferred method for assessing the purity of a fraction is to calculate the specific activity of the fraction, to compare it to the specific activity of the initial extract, and to thus calculate the degree of purity therein, assessed by a "-fold purification number.”
- the actual units used to represent the amount of activity will, of course, be dependent upon the particular assay technique chosen to follow the purification, and whether or not the expressed protein or peptide exhibits a detectable activity.
- Partial purification may be accomplished by using fewer purification steps in combination, or by utilizing different forms of the same general purification scheme. For example, it is appreciated that a cation-exchange column chromatography performed utilizing an HPLC apparatus will generally result in a greater "- fold" purification than the same technique utilizing a low pressure chromatography system. Methods exhibiting a lower degree of relative purification may have advantages in total recovery of protein product, or in maintaining the activity of an expressed protein.
- Affinity chromatography is a chromatographic procedure that relies on the specific affinity between a substance to be isolated and a molecule to which it can specifically bind. This is a receptor-ligand type of interaction.
- the column material is synthesized by covalently coupling one of the binding partners to an insoluble matrix. The column material is then able to specifically adsorb the substance from the solution. Elution occurs by changing the conditions to those in which binding will not occur (e.g., altered pH, ionic strength, temperature, etc.).
- the matrix should be a substance that itself does not adsorb molecules to any significant extent and that has a broad range of chemical, physical and thermal stability.
- the ligand should be coupled in such a way as to not affect its binding properties. The ligand should also provide relatively tight binding. And it should be possible to elute the substance without destroying the sample or the ligand.
- the targeting peptides of the invention can be synthesized in solution or on a solid support in accordance with conventional techniques.
- Various automatic synthesizers are commercially available and can be used in accordance with known protocols. See, for example, Stewart and Young, 1984; Tarn et al, 1983; Merrifield, 1986; Barany and Merrifield, 1979, each incorporated herein by reference. Short peptide sequences, usually from about 6 up to about 35 to 50 amino acids, can be readily synthesized by such methods.
- recombinant DNA technology may be employed wherein a nucleotide sequence which encodes a peptide of the invention is inserted into an expression vector, transformed or transfected into an appropriate host cell, and cultivated under conditions suitable for expression.
- Targeting moieties identified using these methods may be coupled or attached to various substances, including therapeutic or diagnostic agents, for the selective delivery of the conjugate to a desired organ, tissue or cell type in the mouse model system.
- Embodiments of the invention are directed to the treatment of a disease or a disorder, preferably, a cancer.
- the tumor targeting peptide in the present invention may be attached to a molecule; for example, the molecule may be a protein and the peptide may be conjugated or fused to the protein to form a protein conjugate, wherein the protein conjugate is not a naturally occurring protein.
- the peptide may be positioned at a terminus of the protein.
- Said molecule may be a pro-apoptosis agent, an anti-angiogenic agent, a cytokine, a cytotoxic agent, a drug, a chemotherapeutic agent, a hormone, a growth factor, an antibiotic, an antibody or fragment or single chain thereof, a survival factor, an anti-apoptotic agent, a hormone antagonist, an antigen, a peptide, a protein, a diagnostic agent, a radioisotope, or an imaging agent.
- the tumor targeting peptide may be attached to a macromolecular complex, such as a virus, a bacteriophage, a bacterium, a liposome, a microparticle, a magnetic bead, a yeast cell, or a mammalian cell.
- a macromolecular complex such as a virus, a bacteriophage, a bacterium, a liposome, a microparticle, a magnetic bead, a yeast cell, or a mammalian cell.
- said peptide is attached to a virus, such as a lentivirus, papovavirus, adenovirus, retrovirus, AAV, vaccinia virus or herpes virus.
- Said peptide may be attached to a solid support, such as a microtiter dish or microchip.
- Apoptosis or programmed cell death, is an essential process for normal embryonic development, maintaining homeostasis in adult tissues, and suppressing carcinogenesis (Kerr et al, 1972).
- the Bcl-2 family of proteins and ICE-like proteases have been demonstrated to be important regulators and effectors of apoptosis in other systems.
- the Bcl-2 protein plays a prominent role in controlling apoptosis and enhancing cell survival in response to diverse apoptotic stimuli (Bakhshi et al, 1985; Cleary and Sklar, 1985; Cleary et al, 1986; Tsujimoto et al, 1985; Tsujimoto and Croce, 1986).
- the evolutionarily conserved Bcl-2 protein now is recognized to be a member of a family of related proteins, which can be categorized as death agonists or death antagonists.
- Bcl-2 acts to suppress cell death triggered by a variety of stimuli. Also, it now is apparent that there is a family of Bcl-2 cell death regulatory proteins that share in common structural and sequence homologies. These different family members have been shown to either possess similar functions to Bcl-2 ⁇ e.g. , BCI XL , BcIw, BcIs, McI-I, Al, BfI-I) or counteract Bcl-2 function and promote cell death (e.g. , Bax, Bak, Bik, Bim, Bid, Bad, Harakiri).
- pro-apoptosis agent examples include gramicidin; magainin; mellitin; defensin; cecropin; (KLAKLAK) 2 (SEQ ID NO:48); (KLAKKLA) 2 (SEQ ID NO:49); (KAAKKAA) 2
- said pro-apoptosis agent is (KLAKLAK) 2 (SEQ ID NO:48).
- SEQ ID NO:48 may consist of D amino acids.
- the present invention may utilize administration of targeting moieties operatively coupled to anti-angiogenic agents, such as a thrombospondin, an angiostatin, pigment epithelium-derived factor, angiotensin, laminin peptides, fibronectin peptides, plasminogen activator inhibitors, tissue metalloproteinase inhibitors, interferons, interleukin 12, platelet factor 4, IP-10, Gro- ⁇ , thrombospondin, 2-methoxyoestradiol, proliferin-related protein, carboxiamidotriazole, CMlOl, Marimastat, pentosan polysulphate, angiopoietin 2, herbimycin A, PNU145156E, 16K prolactin fragment, Linomide, thalidomide, pentoxifylline, genistein, TNP -470, endostatin, paclitaxel, Docetaxel, polyamines, a protea
- Said molecule may be a cytokine selected from the group consisting of interleukin 1 (IL-I), IL-2, IL-5, IL-10, IL-I l, IL-12, IL-18, interferon- ⁇ (IF- ⁇ ), IF- ⁇ , IF- ⁇ , a tumor necrosis factor, or GM-CSF (granulocyte macrophage colony stimulating factor).
- IL-I interleukin 1
- IL-2 interleukin 2
- IL-5 interleukin 10
- IL-I l interferon- ⁇
- IF- ⁇ interferon- ⁇
- IF- ⁇ IF- ⁇
- IF- ⁇ granulocyte macrophage colony stimulating factor
- GM-CSF granulocyte macrophage colony stimulating factor
- Chemotherapeutic (cytotoxic) agents may be coupled to a targeting peptide of the present invention and used to treat various hyperproliferative or neoplastic disease states, including cancer.
- Chemotherapeutic (cytotoxic) agents of potential use include, but are not limited to, 5-fluorouracil, bleomycin, busulfan, camptothecin, carboplatin, chlorambucil, cisplatin (CDDP), cyclophosphamide, dactinomycin, daunorubicin, doxorubicin, estrogen receptor binding agents, etoposide (VP 16), farnesyl-protein transferase inhibitors, gemcitabine, ifosfamide, mechlorethamine, melphalan, mitomycin, navelbine, nitrosurea, plicomycin, procarbazine, raloxifene, tamoxifen, taxol, temazolomide (an aqueous form of DTIC), transplatinum
- chemotherapeutic agents fall into the categories of alkylating agents, antimetabolites, antitumor antibiotics, corticosteroid hormones, mitotic inhibitors, and nitrosoureas, hormone agents, miscellaneous agents, and any analog or derivative variant thereof.
- Chemotherapeutic agents and methods of administration, dosages, etc. are well known to those of skill in the art (see for example, the “Physicians Desk Reference”, Goodman & Gilman's “The Pharmacological Basis of Therapeutics” and in “Remington's Pharmaceutical Sciences” 15 th ed., pp 1035-1038 and 1570-1580, incorporated herein by reference in relevant parts), and may be combined with the invention in light of the disclosures herein. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
- Alkylating agents are drugs that directly interact with genomic DNA to prevent cells from proliferating. This category of chemotherapeutic drugs represents agents that affect all phases of the cell cycle, that is, they are not phase-specific.
- An alkylating agent may include, but is not limited to, a nitrogen mustard, an ethylenimene, a methylmelamine, an alkyl sulfonate, a nitrosourea or a triazines.
- melphalan examples include but are not limited to: busulfan, chlorambucil, cisplatin, cyclophosphamide (Cytoxan), dacarbazine, ifosfamide, mechlorethamine (mustargen), and melphalan.
- Antimetabolites disrupt DNA and RNA synthesis. Unlike alkylating agents, they specifically influence the cell cycle during S phase. Antimetabolites can be differentiated into various categories, such as folic acid analogs, pyrimidine analogs and purine analogs and related inhibitory compounds. Antimetabolites include but are not limited to, 5-fluorouracil (5-FU), cytarabine (Ara-C), fludarabine, gemcitabine, and methotrexate.
- 5-FU 5-fluorouracil
- Ara-C cytarabine
- fludarabine gemcitabine
- gemcitabine gemcitabine
- methotrexate methotrexate
- Natural products generally refer to compounds originally isolated from a natural source, and identified as having a pharmacological activity. Such compounds, analogs and derivatives thereof may be, isolated from a natural source, chemically synthesized or recombinantly produced by any technique known to those of skill in the art. Natural products include such categories as mitotic inhibitors, antitumor antibiotics, enzymes and biological response modifiers.
- Mitotic inhibitors include plant alkaloids and other natural agents that can inhibit either protein synthesis required for cell division or mitosis. They operate during a specific phase during the cell cycle. Mitotic inhibitors include, for example, docetaxel, etoposide (VP 16), teniposide, paclitaxel, taxol, vinblastine, vincristine, and vinorelbine.
- Mitotic inhibitors include, for example, docetaxel, etoposide (VP 16), teniposide, paclitaxel, taxol, vinblastine, vincristine, and vinorelbine.
- Taxoids are a class of related compounds isolated from the bark of the ash tree, Taxus brevifolia. Taxoids include but are not limited to compounds such as docetaxel and paclitaxel. Paclitaxel binds to tubulin (at a site distinct from that used by the vinca alkaloids) and promotes the assembly of microtubules.
- Vinca alkaloids are a type of plant alkaloid identified to have pharmaceutical activity. They include such compounds as vinblastine (VLB) and vincristine.
- Certain antibiotics have both antimicrobial and cytotoxic activity. These drugs also interfere with DNA by chemically inhibiting enzymes and mitosis or altering cellular membranes. These agents are not phase specific so they work in all phases of the cell cycle.
- cytotoxic antibiotics include, but are not limited to, bleomycin, dactinomycin, daunorubicin, doxorubicin (Adriamycin), plicamycin (mithramycin) and idarubicin.
- Miscellaneous cytotoxic agents that do not fall into the previous categories include, but are not limited to, platinum coordination complexes, anthracenediones, substituted ureas, methyl hydrazine derivatives, amsacrine, L-asparaginase, and tretinoin.
- Platinum coordination complexes include such compounds as carboplatin and cisplatin (cis-
- An exemplary anthracenedione is mitoxantrone.
- An exemplary substituted urea is hydroxyurea.
- An exemplary methyl hydrazine derivative is procarbazine (N- methylhydrazine, MIH).
- the targeting moieties of the present invention may be attached to imaging agents of use for imaging and diagnosis of various diseased organs, tissues or cell types.
- imaging agents are known in the art, as are methods for their attachment to proteins or peptides (see, e.g., U.S. Patents 5,021,236 and 4,472,509, both incorporated herein by reference).
- Certain attachment methods involve the use of a metal chelate complex employing, for example, an organic chelating agent such a DTPA attached to the protein or peptide (U.S. Patent 4,472,509).
- Proteins or peptides also may be reacted with an enzyme in the presence of a coupling agent such as glutaraldehyde or periodate.
- Conjugates with fluorescein markers are prepared in the presence of these coupling agents or by reaction with an isothiocyanate.
- Non-limiting examples of paramagnetic ions of potential use as imaging agents include chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III) and erbium (III), with gadolinium being particularly preferred.
- Ions useful in other contexts, such as X-ray imaging include but are not limited to lanthanum (III), gold (III), lead (II), and especially bismuth (III).
- Radioisotopes of potential use as imaging or therapeutic agents include astatine 211 , 14 carbon, 51 chromium, 36 chlorine, 57 cobalt, 58 cobalt, copper 67 , 152 Eu, gallium 67 , 3 hydrogen, iodine 123 , iodine 125 , iodine 131 , indium 111 , 59 iron, 32 phosphorus, rhenium 186 , rhenium 188 , 75 selenium, 35 sulphur, technicium 99m and yttrium 90 .
- 125 I is often being preferred for use in certain embodiments, and technicium 99m and indium 111 are also often preferred due to their low energy and suitability for long range detection.
- Radioactively labeled proteins or peptides of the present invention may be produced according to well-known methods in the art. For instance, they can be iodinated by contact with sodium or potassium iodide and a chemical oxidizing agent such as sodium hypochlorite, or an enzymatic oxidizing agent, such as lactoperoxidase.
- a chemical oxidizing agent such as sodium hypochlorite
- an enzymatic oxidizing agent such as lactoperoxidase.
- Proteins or peptides according to the invention may be labeled with technetium-" 1 " by ligand exchange process, for example, by reducing pertechnate with stannous solution, chelating the reduced technetium onto a Sephadex column and applying the peptide to this column or by direct labeling techniques, e.g., by incubating pertechnate, a reducing agent such as SNCl 2 , a buffer solution such as sodium-potassium phthalate solution, and the peptide.
- Intermediary functional groups that are often used to bind radioisotopes that exist as metallic ions to peptides are diethylenetriaminepenta-acetic acid (DTPA) and ethylene diaminetetra-acetic acid (EDTA).
- fluorescent labels including rhodamine, fluorescein isothiocyanate and renographin.
- the claimed proteins or peptides may be linked to a secondary binding ligand or to an enzyme (an enzyme tag) that will generate a colored product upon contact with a chromogenic substrate.
- suitable enzymes include urease, alkaline phosphatase, (horseradish) hydrogen peroxidase and glucose oxidase.
- Preferred secondary binding ligands are biotin and avidin or streptavidin compounds. The use of such labels is well known to those of skill in the art in light and is described, for example, in U.S. Patents 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149 and 4,366,241; each incorporated herein by reference.
- a targeting moiety may be operatively coupled to a nanoparticle.
- Nanoparticles include, but are not limited to colloidal gold and silver nanoparticles.
- Metal nanoparticles exhibit colors in the visible spectral region. It is believed that these colors are the result of excitation of surface plasmon resonances in the metal particles and are extremely sensitive to particles' sizes, shapes, and aggregation state; dielectric properties of the surrounding medium; adsorption of ions on the surface of the particles (For examples see U.S. Patent Application 20040023415, which is incorporated herein by reference).
- Cross linkers may also be included in a fusion protein or other construct comprising a CRKL targeting peptide; for example, corss linkers may be useful for conjugating a targeting peptide to a liposome or a tharapeutic compound.
- Bifunctional cross- linking reagents have been extensively used for a variety of purposes including preparation of affinity matrices, modification and stabilization of diverse structures, identification of ligand and receptor binding sites, and structural studies. Homobifunctional reagents that carry two identical functional groups proved to be highly efficient in inducing cross-linking between identical and different macromolecules or subunits of a macromolecule, and linking of polypeptide ligands to their specific binding sites.
- Heterobifunctional reagents contain two different functional groups. By taking advantage of the differential reactivities of the two different functional groups, cross-linking can be controlled both selectively and sequentially.
- the bifunctional cross-linking reagents can be divided according to the specificity of their functional groups, e.g., amino, sulfhydryl, guanidino, indole, carboxyl specific groups. Of these, reagents directed to free amino groups have become especially popular because of their commercial availability, ease of synthesis and the mild reaction conditions under which they can be applied.
- a majority of heterobifunctional cross-linking reagents contains a primary amine-reactive group and a thiol-reactive group.
- ligands can be covalently bound to liposomal surfaces through the cross- linking of amine residues.
- Liposomes in particular, multilamellar vesicles (MLV) or unilamellar vesicles such as microemulsified liposomes (MEL) and large unilamellar liposomes (LUVET), each containing phosphatidylethanolamine (PE), have been prepared by established procedures.
- MLV multilamellar vesicles
- MEL microemulsified liposomes
- LVET large unilamellar liposomes
- PE in the liposome provides an active functional residue, a primary amine, on the liposomal surface for cross-linking purposes.
- Ligands such as epidermal growth factor (EGF) have been successfully linked with PE-liposomes. Ligands are bound covalently to discrete sites on the liposome surfaces. The number and surface density of these sites are dictated by the liposome formulation and the liposome type. The liposomal surfaces may also have sites for non-covalent association.
- cross-linking reagents have been studied for effectiveness and biocompatibility.
- Cross-linking reagents include glutaraldehyde (GAD), bifunctional oxirane (OXR), ethylene glycol diglycidyl ether (EGDE), and a water soluble carbodiimide, preferably l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC).
- GAD glutaraldehyde
- OXR bifunctional oxirane
- EGDE ethylene glycol diglycidyl ether
- EDC water soluble carbodiimide
- heterobifunctional cross-linking reagents and methods of using the cross-linking reagents are described (U.S. Patent 5,889,155, specifically incorporated herein by reference in its entirety).
- the cross-linking reagents combine a nucleophilic hydrazide residue with an electrophilic maleimide residue, allowing coupling in one example, of aldehydes to free thiols.
- the cross-linking reagent can be modified to crosslink various functional groups.
- Nucleic acids according to the present invention may encode a targeting peptide, a targeting antibody, a therapeutic polypeptide a fusion protein or other protein or peptide.
- the nucleic acid may be derived from genomic DNA, complementary DNA (cDNA) or synthetic DNA.
- nucleic acid as used herein includes single-stranded and double-stranded molecules, as well as DNA, RNA, chemically modified nucleic acids and nucleic acid analogs. It is contemplated that a nucleic acid within the scope of the present invention may be of almost any size, determined in part by the length of the encoded protein or peptide.
- targeting peptides and fusion proteins may be encoded by any nucleic acid sequence that encodes the appropriate amino acid sequence.
- the design and production of nucleic acids encoding a desired amino acid sequence is well known to those of skill in the art, using standardized codon tables.
- the codons selected for encoding each amino acid may be modified to optimize expression of the nucleic acid in the host cell of interest.
- Targeting peptides may, in certain embodiments, be coupled to a gene therapy vector to selectively or preferentially target cells expressing CRKL on the cell surface, such as certain tumor cells.
- a gene therapy vector comprises a virus. The ability of certain viruses to enter cells via receptor-mediated endocytosis, to integrate into host cell genome or be maintained episomally, and express viral genes stably and efficiently have made them attractive candidates for the transfer of foreign genes into mammalian cells (Ridgeway, 1988; Nicolas and Rubinstein, 1988.; Baichwal and Sugden, 1986; Temin, 1986).
- Preferred gene therapy vectors are generally viral vectors.
- DNA viruses used as gene therapy vectors include the papovaviruses (e.g., simian virus 40, bovine papilloma virus, and polyoma) (Ridgeway, 1988; Baichwal and Sugden, 1986) and adenoviruses (Ridgeway, 1988; Baichwal and Sugden, 1986).
- papovaviruses e.g., simian virus 40, bovine papilloma virus, and polyoma
- adenoviruses Rosgeway, 1988; Baichwal and Sugden, 1986.
- adenovirus expression vector is meant to include, but is not limited to, constructs containing adenovirus sequences sufficient to (a) support packaging of the construct and (b) to express an antisense or a sense polynucleotide that has been cloned therein.
- Adenovirus vectors have been used in eukaryotic gene expression (Levrero et al, 1991; Gomez -Foix et al, 1992) and vaccine development (Grunhaus and Horwitz, 1992; Graham and Prevec, 1991).
- Studies in administering recombinant adenovirus to different tissues include trachea instillation (Rosenfeld et al, 1991; Rosenfeld et al, 1992), muscle injection (Ragot et al, 1993), peripheral intravenous injections (Herz and Gerard, 1993) and stereotactic innoculation into the brain (Le Gal La Salle et al, 1993).
- certain advantages may be gained from coupling therapeutic molecules or substances to tumor targeting peptides that target to cells expressing and bearing CRKL on the surface, such as tumor cells.
- moieties that home to tumor vasculature have been coupled to cytotoxic drugs or proapoptotic peptides to yield compounds were more effective and less toxic than the parental compounds in experimental models of mice bearing tumor xenografts (Arap et al, 1998; Ellerby et al, 1999).
- the insertion of the RGD-4C peptide into a surface protein of an adenovirus has produced an adenoviral vector that may be used for tumor targeted gene therapy (Arap et al, 1998).
- the peptide motif allows for cell targeting, for instance, by comprising a targeting moiety of the invention, and/or a ligand for a cell surface binding site.
- the peptide motif optionally can comprise other elements of use in cell targeting ⁇ e.g. , a single-chain antibody sequence).
- the peptide binding motif may be generated by the insertion, and may comprise, for instance, native and nonnative sequences, or may be entirely made up of nonnative sequences.
- the peptide motif that results from the insertion of the nonnative amino acid sequence into the chimeric fiber protein can be either a high affinity peptide (i.e., one that binds its cognate binding site, e.g., CRKL, when provided at a relatively low concentration) or a low affinity peptide (i.e., one that binds its cognate binding site, e.g., CRKL, when provided at a relatively high concentration).
- the resultant peptide motif is a high affinity motif, particularly one that has a high affinity for its cognate binding site due to its constraint within the adenovirus fiber protein.
- a nucleic acid encoding protein of interest is inserted into the viral genome in the place of certain viral sequences to produce a virus that is replication-defective.
- Retroviral vectors are capable of infecting a broad variety of cell types. However, integration and stable expression require the division of host cells (Paskind et al, 1975).
- viral vectors may be employed as targeted gene therapy vectors.
- Vectors derived from viruses such as vaccinia virus (Ridgeway, 1988; Baichwal and Sugden, 1986), adeno-associated virus (AAV) (Ridgeway, 1988; Baichwal and Sugden, 1986; Hermonat and Muzycska, 1984), and herpes viruses may be employed.
- gene therapy construct may be entrapped in a liposome. Liposome-mediated nucleic acid delivery and expression of foreign DNA in vitro has been very successful.
- Wong et ah (1980) demonstrated the feasibility of liposome-mediated delivery and expression of foreign DNA in cultured chick embryo, HeLa, and hepatoma cells.
- Nicolau et ah, (1987.) accomplished successful liposome-mediated gene transfer in rats after intravenous injection.
- Gene therapy vectors of the invention may comprise various transgenes, which are typically encoded DNA or RNA of an expression vector. Gene therapy may be used for the expression of a therapeutic gene, expression of VEGFR- 1/NRP-l to enhance neo-vascularization or for the inhibition of VEGFR- 1/NRP-l expression for the treatment of disease states associated with neo-vascularization.
- DNA may be in form of cDNA, in vitro polymerized DNA, plasmid DNA, parts of a plasmid DNA, genetic material derived from a virus, linear DNA, vectors (Pl, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, recombinant DNA, chromosomal DNA, an oligonucleotide, anti-sense DNA, or derivatives of these groups.
- RNA may be in the form of oligonucleotide RNA, tRNA (transfer RNA), snRNA (small nuclear RNA), rRNA (ribosomal RNA), mRNA (messenger RNA), in vitro polymerized RNA, recombinant RNA, chimeric sequences, anti-sense RNA, siRNA (small interfering RNA), ribozymes, or derivatives of these groups.
- An anti-sense polynucleotide is a polynucleotide that interferes with the function of DNA and/or RNA.
- Antisense polynucleotides include, but are not limited to: morpholinos, 2'-O-methyl polynucleotides, DNA, RNA and the like.
- SiRNA comprises a double stranded structure typically containing 15-50 base pairs and preferably 21-25 base pairs and having a nucleotide sequence identical or nearly identical to an expressed target gene or RNA within the cell. Interference may result in suppression of expression.
- DNA and RNA may be single, double, triple, or quadruple stranded.
- compositions of the present invention comprise an effective amount of one or more said tumor targeting peptide or additional agent dissolved or dispersed in a pharmaceutically acceptable carrier.
- pharmaceutically acceptable refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal, such as, for example, a human, as appropriate.
- the preparation of an pharmaceutical composition that contains at least one tumor targeting peptide in the present invention or additional active ingredient will be known to those of skill in the art in light of the present disclosure, as exemplified by Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, incorporated herein by reference.
- preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biological Standards.
- pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, such like materials and combinations thereof, as would be known to one of ordinary skill in the art (see, for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329, incorporated herein by reference). Except insofar as any conventional carrier is incompatible with the active ingredient, its use in the pharmaceutical compositions is contemplated.
- the composition may comprise different types of carriers depending on whether it is to be administered in solid, liquid or aerosol form, and whether it need to be sterile for such routes of administration as injection.
- the present invention can be administered intravenously, intradermally, transdermally, intrathecally, intraarterially, intraperitoneally, intranasally, intravaginally, intrarectally, topically, intramuscularly, subcutaneously, mucosally, orally, topically, locally, inhalation (e.g., aerosol inhalation), injection, infusion, continuous infusion, localized perfusion bathing target cells directly, via a catheter, via a lavage, in cremes, in lipid compositions (e.g., liposomes), or by other method or any combination of the forgoing as would be known to one of ordinary skill in the art (see, for example, Remington: The Science and Practice of Pharmacy, 21th Ed. Mack Printing Company, 2005, incorporated herein by reference).
- the composition of the present invention suitable for administration is provided in a pharmaceutically acceptable carrier with or without an inert diluent.
- the carrier should be assimilable and includes liquid, semi-solid, i.e., pastes, or solid carriers. Except insofar as any conventional media, agent, diluent or carrier is detrimental to the recipient or to the therapeutic effectiveness of a the composition contained therein, its use in administrable composition for use in practicing the methods of the present invention is appropriate.
- carriers or diluents include fats, oils, water, saline solutions, lipids, liposomes, resins, binders, fillers and the like, or combinations thereof.
- composition may also comprise various antioxidants to retard oxidation of one or more component. Additionally, the prevention of the action of microorganisms can be brought about by preservatives such as various antibacterial and antifungal agents, including but not limited to parabens (e.g., methylparabens, propylparabens), chlorobutanol, phenol, sorbic acid, thimerosal or combinations thereof.
- parabens e.g., methylparabens, propylparabens
- chlorobutanol phenol
- sorbic acid thimerosal or combinations thereof.
- the composition is combined with the carrier in any convenient and practical manner, i.e., by solution, suspension, emulsification, admixture, encapsulation, absorption and the like. Such procedures are routine for those skilled in the art.
- an "anti-cancer” agent is capable of negatively affecting cancer in a subject, for example, by killing cancer cells, inducing apoptosis in cancer cells, reducing the growth rate of cancer cells, reducing the incidence or number of metastases, reducing tumor size, inhibiting tumor growth, reducing the blood supply to a tumor or cancer cells, promoting an immune response against cancer cells or a tumor, preventing or inhibiting the progression of cancer, or increasing the lifespan of a subject with cancer.
- these other compositions would be provided in a combined amount effective to kill or inhibit proliferation of the cell.
- This process may involve contacting the cells with the expression construct and the agent(s) or multiple factor(s) at the same time. This may be achieved by contacting the cell with a single composition or pharmacological formulation that includes both agents, or by contacting the cell with two distinct compositions or formulations, at the same time, wherein one composition includes the expression construct and the other includes the second agent(s).
- Tumor cell resistance to chemotherapy and radiotherapy agents represents a major problem in clinical oncology.
- One goal of current cancer research is to find ways to improve the efficacy of chemo- and radiotherapy by combining it with gene therapy.
- the herpes simplex -thymidine kinase (HS-tK) gene when delivered to brain tumors by a retroviral vector system, successfully induced susceptibility to the antiviral agent ganciclovir (Culver, et ah, 1992).
- the tumor targeting peptide could be used similarly in conjunction with chemotherapeutic, radiotherapeutic, or immunotherapeutic intervention, in addition to other pro-apoptotic or cell cycle regulating agents.
- the gene therapy may precede or follow the other agent treatment by intervals ranging from minutes to weeks.
- the other agent and expression construct are applied separately to the cell, one would generally ensure that a significant period of time did not expire between the time of each delivery, such that the agent and expression construct would still be able to exert an advantageously combined effect on the cell.
- the tumor targeting peptide is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoe)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N
- Cancer therapies also include a variety of combination therapies with both chemical and radiation based treatments.
- Combination chemotherapies include, for example, cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide (VP 16), tamoxifen, raloxifene, estrogen receptor binding agents, taxol, gemcitabien, navelbine, farnesyl-protein tansferase inhibitors, transplatinum, 5-fluorouracil, vincristin, vinblastin and methotrexate, or any analog or derivative variant of the foregoing.
- CDDP cisplatin
- carboplatin carboplatin
- DNA damaging factors include what are commonly known as ⁇ -rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells.
- Other forms of DNA damaging factors are also contemplated such as microwaves and UV-irradiation. It is most likely that all of these factors effect a broad range of damage on DNA, on the precursors of DNA, on the replication and repair of
- Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens.
- Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.
- contacted and “exposed,” when applied to a cell are used herein to describe the process by which a therapeutic construct and a chemotherapeutic or radiotherapeutic agent are delivered to a target cell or are placed in direct juxtaposition with the target cell.
- both agents are delivered to a cell in a combined amount effective to kill the cell or prevent it from dividing.
- Immunotherapeutics generally, rely on the use of immune effector cells and molecules to target and destroy cancer cells.
- the immune effector may be, for example, an antibody specific for some marker on the surface of a tumor cell.
- the antibody alone may serve as an effector of therapy or it may recruit other cells to actually effect cell killing.
- the antibody also may be conjugated to a drug or toxin (chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.) and serve merely as a targeting agent.
- the effector may be a lymphocyte carrying a surface molecule that interacts, either directly or indirectly, with a tumor cell target.
- Various effector cells include cytotoxic T cells and NK cells.
- Immunotherapy could be used as part of a combined therapy, in conjunction with a therapy with the tumor targeting peptide.
- the general approach for combined therapy is discussed below.
- the tumor cell must bear some marker that is amenable to targeting, i.e., is not present on the majority of other cells.
- Common tumor markers include carcinoembryonic antigen, prostate specific antigen, urinary tumor associated antigen, fetal antigen, tyrosinase (p97), gp68, TAG-72, HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor, laminin receptor, erb B and pl55.
- the secondary treatment is a gene therapy in which a therapeutic polynucleotide is administered before, after, or at the same time a first therapeutic agent comprising an isolated tumor targeting peptide containing a CRKL binding motif. Delivery of the therapeutic agent in conjuction with a second vector encoding one of the following gene products will have a combined anti-hyperproliferative effect on target tissues.
- Curative surgery is a cancer treatment that may be used in conjunction with other therapies, such as the treatment of the present invention, chemotherapy, radiotherapy, hormonal therapy, gene therapy, immunotherapy and/or alternative therapies.
- Curative surgery includes resection in which all or part of cancerous tissue is physically removed, excised, and/or destroyed.
- Tumor resection refers to physical removal of at least part of a tumor.
- treatment by surgery includes laser surgery, cryosurgery, electrosurgery, and miscopically controlled surgery (Mohs' surgery). It is further contemplated that the present invention may be used in conjunction with removal of superficial cancers, precancers, or incidental amounts of normal tissue.
- a cavity may be formed in the body.
- Treatment may be accomplished by perfusion, direct injection or local application of the area with an additional anti-cancer therapy.
- Such treatment may be repeated, for example, every 1, 2, 3, 4, 5, 6, or 7 days, or every 1, 2, 3, 4, and 5 weeks or every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.
- These treatments may be of varying dosages as well.
- agents may be used in combination with the present invention to improve the therapeutic efficacy of treatment.
- additional agents include immunomodulatory agents, agents that affect the upregulation of cell surface receptors and GAP junctions, cytostatic and differentiation agents, inhibitors of cell adehesion, or agents that increase the sensitivity of the hyperproliferative cells to apoptotic inducers.
- Immunomodulatory agents include tumor necrosis factor; interferon alpha, beta, and gamma; IL-2 and other cytokines; F42K and other cytokine analogs; or MIP-I, MIP- lbeta, MCP-I, RANTES, and other chemokines.
- cell surface receptors or their ligands such as Fas / Fas ligand, DR4 or DR5 / TRAIL would potentiate the apoptotic inducing abililties of the present invention by establishment of an autocrine or paracrine effect on hyperproliferative cells. Increases intercellular signaling by elevating the number of GAP junctions would increase the anti- hyperproliferative effects on the neighboring hyperproliferative cell population.
- cytostatic or differentiation agents can be used in combination with the present invention to improve the anti-hyerproliferative efficacy of the treatments. Inhibitors of cell adehesion are contemplated to improve the efficacy of the present invention.
- cell adhesion inhibitors are focal adhesion kinase (FAKs) inhibitors and Lovastatin. It is further contemplated that other agents that increase the sensitivity of a hyperproliferative cell to apoptosis, such as the antibody c225, could be used in combination with the present invention to improve the treatment efficacy.
- FAKs focal adhesion kinase
- Lovastatin agents that increase the sensitivity of a hyperproliferative cell to apoptosis
- Hormonal therapy may also be used in conjunction with the present invention or in combination with any other cancer therapy previously described. The use of hormones may be employed in the treatment of certain cancers such as breast, prostate, ovarian, or cervical cancer to lower the level or block the effects of certain hormones such as testosterone or estrogen. This treatment is often used in combination with at least one other cancer therapy as a treatment option or to reduce the risk of metastases.
- Example 1 Selection of tumor-homing peptides in vivo
- a phage display random peptide library (Arap et al., 1998; Pasqualini and Ruoslahti, 1996; Pasqualini et al., 2001) was administered intravenously into nu/nu (nude) mice bearing human DU 145 -derived prostate cancer xenografts and the tumors were recovered after a 24 hour circulation timeframe. After three rounds of selection, an enriched population of tumor-targeting phage was recovered and the DNA corresponding to the peptide inserts displayed by individual phage clones was sequenced (Table 1).
- the dominant peptide was selected, amino acid sequence YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) for functional characterization.
- SEQ ID NO:1 amino acid sequence for functional characterization.
- the tumor-targeting specificity of the YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) peptide was evaluated in vivo in DU145- derived tumor-bearing mice.
- SEQ ID NO: 1 systemic intravenous administration of the individual YRCTLNSPFFWEDMTHECHA (SEQ ID NO : 1 )-displaying phage clone
- marked homing to tumor xenografts was observed (insertless phage served as a negative control) with no or barely detectable phage localization noted in several control organs.
- DU 145 prostate cancer cells were also targeted in vitro by using an aqueous to organic phase separation assay (Giordano et al., 2001) and a phage-based immunofluorescence assay (KS 1767 cells) and it was found that phage displaying the peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO: 1) binds to tumor cell surfaces much more than a negative control phage displaying no peptide insert (FIG. IA, B).
- anti-CRKL (Santa Cruz, Cell Signaling, Epitomics or Upstate Biotechnology), anti-phospho-CRKL (Cell Signaling), anti- ⁇ i integrin (Chemicon or BD Transduction Laboratories), anti-ILHR (Santa Cruz), anti- ⁇ 3 and anti- ⁇ s integrins47, anti-EGFR48, anti-grb2 (Santa Cruz), anti-alpha6 integrin (Chemicon), anti-AHSG/Feutin A (R&D Systems), pre-immune serum (Jackson Laboratory), anti-FAK (Upstate), anti-Histone Hl (Santa Cruz), anti-phospho-paxillin (Cell Signaling), anti-phospho-130Cas (Santa Cruz), anti-phospho-Erkl/2 (Cell Signaling or Biosource), anti- phospho-Elk-1 (Cell Signaling), anti-His (Santa Cruz), anti-gst (Santa Cruz), anti-gst (Santa
- a random phage library displaying an insert with the general arrangement X 2 CX 12 CX 2 (C, cysteine; X, any residue) was systemically administered (tail vein) into athymic nude mice bearing tumor xenografts derived from human DU 145 prostate cancer cells and allowed to circulate for 24 hours.
- mice were placed under deep anesthesia, tumor xenografts excised, weighed, and the bound phage population was recovered and processed (Arap et ⁇ l, 1998; Arap et ⁇ l, 2004; Pasqualini and Ruoslahti, 1996; Pasqualini et ⁇ l, 2001). Three serial rounds of in vivo selection were performed.
- a tumor-homing peptide sequence mimics a regulatory integrin extracellular domain [00128]
- a similarity search of YRCTLNSPFFWEDMTHECHA SEQ ID NO:1
- SEQ ID NO:1 YRCTLNSPFFWEDMTHECHA
- the dominant peptide sequence YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) had similarity to the plexin-semaphorin-integrin (PSI) extracellular domain (residues 26-78) of the ⁇ i integrin chain (FIG. 2A; moreover, it was found that other selected peptides also appeared within the same region (FIG. 2C). It was then determined whether the similarity of the selected peptide sequence YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) was specific for the PSI domain of the ⁇ i integrin sequence or common to other known integrin ⁇ chains. After fit analysis, and molecular modeling, it was concluded that the sequence identity between YRCTLNSPFFWEDMTHECHA (SEQ ID NO: 1) and the PSI domain of ⁇ i integrin was indeed the best alignment (FIG. 2B).
- the program calculates similarity based on a predefined residue window size between an affinity selected peptide sequence and the target protein sequence from N- to C- protein terminus in one-residue shifts.
- the peptide-protein similarity scores for each residue were calculated based on a BLOSUM62 amino acid substitution matrix modified to adjust for rare amino acid representation. Thresholds were set at least 4 identical residues between the peptide and the protein segment to discriminate significant similarities from nonspecific background matches.
- a cytoplasmic adapter protein serves as a receptor for the PSI domain- like tumor-homing peptide
- the integrin PSI domain has been well characterized for its regulatory activity (Shi et ⁇ l., 2005; Mould et ⁇ l., 2005; Arnaout et ⁇ l., 2005; Juliano et ⁇ l., 2004). Given the selection results, it seems that the PSI domain might also function as a ligand- receptor binding site within ⁇ i integrins. In view of this, affinity chromatography was used to identify binding partners to the tumor-homing peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1).
- a DU 145 -derived cell extract was pre-cleared through a control peptide column and then the pre-cleared extract was passed through the tumor-homing YRCTLNSPFFWEDMTHECHA (SEQ ID NO: 1) peptide column followed by an acidic elution. A specific gel band corresponding to an ⁇ 40-KDa protein was detected that eluted from the column.
- the ⁇ i integrin PSI-derived peptide NSTFLQEGMPTSA (SEQ ID NO:23) bound to rCRKL, rCRKL-SH3 (N) domain, and rCRKL-SH3 (C) domain (FIG. 2B).
- cyclic and linear peptide sequences from the PSI domain were generated and it was found that the cyclic disulfide bonds present in the PSI domain are not essential for the binding to CRKL (FIG. 3C).
- the interaction between the SH3 (C) domain and the tumor-homing peptide is specifically inhibited by the corresponding synthetic peptides and by the phage clone itself (FIG. 3D).
- Standard peptide affinity columns were made by EDC and DADPA immobilization resin (Pierce).
- DUl 45 tumor cell extracts were prepared and first passed through a non-specific control peptide column followed by the tumor-homing peptide column. Columns were washed extensively, then eluted with glycine (pH 2.2), and analyzed by SDS-PAGE. Then, the gels were Coomassie-stained. A band of ⁇ 40KDa was detected and excised for protein sequencing by mass spectrometry at the UTMDACC Proteomic Core Facility. The protein was identified as CRKL.
- Affinity purification of CRKL from serum- free condition medium was performed and confirmed by using recombinant gst-tag fusion protein expressing either the tumor-homing peptide or a mutant control peptide (Pro— »Ala). Approximately 200 ml of serum-free condition medium (48 hours of culturing) was concentrated for the affinity purification. The recombinant fusion proteins were coupled to gst-resin beads and loaded on a column. The concentrated serum-free condition medium was added to the coupled columns and incubated overnight. After several washes the bound CRKL was eluted for Western blotting. The blot was probed with anti-gst and anti-CRKL antibodies.
- YRCTLNSPFFWEDMTHECHA wild-type tumor-homing phage
- SEQ ID NO: 1 scrambled phage
- YRFCTSPFHEWHLENTDMCA SEQ ID NO:26
- YRECTDSPHEFHLWNTMCAF SEQ ID NO:27
- YRCETD SPHEFHLWNTMC AF SEQ ID NO:29
- YRCETDSPHEFHLWNTFCAM mutant phage
- microtiter-wells were coated at 4°C overnight with 250 ⁇ g/ml of recombinant gst- SH3 domains (CKRL-Dl, CKRL-D2, grb2-Dl, grb2-D2, Lyn, src; Pronomics), rCKRL protein, and negative controls gst or BSA.
- SH3 (C) mutant binding studies 2 ⁇ g/ml of His-tag recombinant wild-type SH3-C and mutant SH3 (C) domains were coated.
- phage displaying scrambled peptide sequences and mutants designed and constructed (Pro— »Ala and Phe-Phe-Trp— »Ala-Ala-Ala) from the selected CRKL binding tumor homing phage peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1).
- Scrambled peptide sequences (YRFCTSPFHEWHLENTDMCA (SEQ ID NO:26),
- YRECTDSPHEFHLWNTMCAF SEQ ID NO:27
- YRCETDSPHEFHLWNTMCAF SEQ ID NO:29
- YRCETD SPHEFHLWNTFC AM SEQ ID NO:30
- mutants YRCTLNSAFFWEDMTHECHA (SEQ ID NO:25) and YRCTLNSPAAAEDMTHECHA (SEQ ID NO:28)
- native PSI-derived phage cloned into the Sf ⁇ l-digested fUSE5 vectors (Smith and Scott, 1993).
- each of the synthetic oligonucleotide templates corresponding to the displayed peptides were converted to double-stranded DNA by PCR amplification with the primer set 5' GTGAGCCGGCTGCCC 3' (SEQ ID NO:68) and 5' TTCGGCCCCAGCGGC 3' (SEQ ID NO: 69) (Sigma-Genosys) and 2.5 U of Taq-DNA polymerase (Promega) in 20 ⁇ l as follows: 94°C for 2 min, followed by 35 cycles at 94°C for 30 s, 60 0 C for 30 s, and 72°C for 30 s, followed by 72°C for 5 min.
- Double-stranded DNA sequences that contained BgII restriction sites in the insert-flanking regions were purified by using a QIAquick nucleotide removal kit (Qiagen) and eluted. Oligonucleotides were digested with BgII for 2 hour at 37°C, re-purified, and ligated into Sfil-digested fUSE5 vector. The phage clones generated were PCR amplified to verify the correct insertion and nucleotide sequence. The individual phage clones were tested in phage binding assays.
- SEQ ID NO:1 PSI-derived peptide
- NSTFLQEGMPTSA PSI-derived peptide
- SEQ ID NO:23 PSI-derived peptide
- the following recombinant His-tag proteins were added to the coated plates: rCRKL, rCRKL-SH2 domain, rCRKL-SH3 (N) domain, and rCRKL-SH3 (C) domain.
- An unrelated control protein ( ⁇ 2-Heremans-Schmid glycoprotein; AHSG) served as a negative control. The mixtures were incubated, washed, and labeled with the appropriate antibodies.
- HRP horse-radish peroxidase
- the non-permeabilized cells were incubated with rabbit anti-fd bacteriophage antibody (Sigma) for 2 hours at room temperature followed by 1 hour incubation with Cy3-labebled anti-rabbit IgG antibody (Jackson ImmunoResearch). Cells were again fixed with 4% paraformaldehyde and mounted in the presence of DAPI (Vector Laboratories) and images were acquired with an Olympus fluorescence microscope.
- rabbit anti-fd bacteriophage antibody Sigma
- Cy3-labebled anti-rabbit IgG antibody Jackson ImmunoResearch
- CRKL can be purified from DU 145 serum- free condition medium, and that a control column prepared with a mutant form of the tumor-homing peptide no longer binds to CRKL at detectable levels under identical experimental conditions.
- a control column prepared with a mutant form of the tumor-homing peptide no longer binds to CRKL at detectable levels under identical experimental conditions.
- CRKL and ⁇ i integrin form a cell surface complex; in contrast, control antibodies raised against unrelated transmembrane receptors including anti- ILI l receptor, anti-EGF receptor, or other integrins (anti- ⁇ 3 and anti- ⁇ 5 ) showed no association with CRKL or ⁇ i integrin.
- CRKL mutants in the SH3 domain were generated by PCR mutagenesis. Primer sets were designed to remove 60 bp ( ⁇ l and ⁇ 2) and 54 bp
- Example 5 Molecular imaging shows that CRKL can localize outside cells
- Membrane fractionation was performed as described (Mintz et ⁇ l., 2003). Immunoblots were probed with the appropriate antibodies as indicated. Confocal images were acquired on an LSM 510 (Carl Zeiss) confocal microscope. DUl 45 cells were grown on fibronectin-coated slides, fixed with 4% paraformaldehyde (PFA) and labeled with the appropriate antibodies (polyclonal CRKL antibody and monoclonal AHSG antibody). The electron microscopy images were acquired at the High Resolution Electron Microscopy Core Facility (JSM 5900 scanning and JEM 1010 transmission electron microscopes). Gold nanoparticle antibody- conjugates were prepared by mixing 20-25 ⁇ m or 40-45 ⁇ m gold in sodium borate.
- Gold- coupled nanoparticles were verified by TEM analysis.
- DU145 cells were labeled on ice with appropriate antibodies (monoclonal anti-CRKL, anti- ⁇ i integrin, and anti-AHSG antibodies) followed by secondary conjugated-fluorescent antibodies and analyzed by FACS.
- Appropriate antibodies monoclonal anti-CRKL, anti- ⁇ i integrin, and anti-AHSG antibodies
- secondary conjugated-fluorescent antibodies and analyzed by FACS.
- CRKL Since CRKL does not have a classic transmembrane domain, studies to evaluate whether or not CRKL is secreted from tumor cells were undertaken. Results showed that DU 145 cells cultured in a serum- free medium do secrete the unphosphorylated form of CRKL. In contrast, CRKL was not detected in control cell media as shown by immunoprecipitation either with anti-CRKL or with several control antibodies. To assess the generality of these observations, a panel of tumor cell lines in serum- free media were studied and found that they also secrete unphosphorylated CRKL (FIG. 7A), thus indicating that this phenomenon is unlikely to be cell type-specific.
- CRKL knockdown by siRNA technology was empolyed; again, it was found that significant reductions in cell proliferation, adhesion, and migration resulted when CRKL expression is reduced (FIG. 8A-C). As an additional control, it was shown that the decrease in cell proliferation could be rescued by exogenous CRKL (FIG. 8D) and only background levels of apoptosis (less than 1%) were detected in the CRKL siRNA knockdown cells or in serum-free medium cultured cells. CRKL siRNA knockdown in cells also was found to reduced binding of the tumor-homing phage to the cells, suggesting that the tumor-homing phage may bind through secreted CRKL.
- results shown provided here do not exclude the possibility that cell death—either as part of clonal selection during malignant progression or after cytotoxic chemotherapy—could also generate extracellular CRKL within the tumor microenvironment.
- binding assays were designed to detail the biochemical interactions among CRKL, P 1 integrins, and the PSI domain-mimic peptide.
- CRKL is implicated in both MAP kinase and integrin- mediated pathways (Li et al, 2003; Uemura et al, 1999), phosphorylated proteins in these two pathways were examined. Several phosphorylated proteins including paxillin, pl30Cas, Erkl, Erk2, and Elkl, and even CRKL itself were found when recombinant CRKL was added to tumor cells in vitro.
- CRKL mRNA accession # NM_005207
- ⁇ i integrin mRNA accession # NM_002211
- control siRNAs were purchased (Santa Cruz, Ambion and
- siRNA oligonucleotides sequences and corresponding manufacturers are summarized in (Table 3). Oligofectamine (Invitrogen) or DharmaFect (Dharmacon) were used to transfect siRNAs into DU 145 cells (1-2x10 5 cells per well). The transfected cells were incubated for 48-72 hours prior to processing. No cell death or apoptosis was observed in the transfected cells as determined by Annexin-V staining (Roche). Transfected cells were harvested and lysed in the presence of protease inhibitors. The tumor-homing phage binding activity was also examined in the CRKL knockdown cells.
- CRKL recombinant CRKL
- Exogenous His-tag recombinant CRKL protein 400 ⁇ g/ml
- control proteins EGF, 200 ⁇ g/ml
- MIF 200 ⁇ g/ml
- PMA 300 ⁇ g/ml
- Equal amounts of protein were loaded and resolved by SDS-PAGE followed by Western blot analysis with the appropriate antibodies.
- Cell proliferation assays were performed with WST-I (Roche).
- Cell migration assays were performed in a Boyden chamber assay (Corning).
- GUCGUAUUGUCAAAGAGUATT SEQ ID NO:54
- GUAGCAGACAACACACAAATT SEQ ID NO:55
- CAGCAGACCUAGAAAUGUATT SEQ ID NO:56
- GGUAUCCAAGCCCACCAAUTT (SEQ ID NO:61)
- GGAUGAAUAUAAAUGGCCATT (SEQ ID NO:62)
- phage constructs displaying either the selected CRKL-binding peptide or a panel of control (mutant or scrambled) peptides were designed and produced. Phage clones were tested in human tumor xenografts (Kaposi sarcoma KS 1767 cells and prostate carcinoma DU 145 cells) and an isogenic mouse tumor model (EF43-FGF4 mammary carcinoma). Marked and specific tumor homing was observed after systemic administration of the CRKL-binding phage; in contrast, control constructs showed no localization to tumors (FIG. 9A-D). As shown in FIG.
- mutanted peptide sequences P- >A (YRCTLNSAFFWEDMTHECHA; SEQ ID NO:25), scramble 1 (YRFCTSPFHEWHLENTDMCA; SEQ ID NO:26) and scramble 2 (YRECTDSPHEFHLWNTMCAF; SEQ ID NO:27) deomnstrated no tumor targeting Additional studies with peptides comprising a FFW->AAA mutation adjacent to the pro line residue in SEQ ID NO:1 (YRCTLNSPAAAEDMTHECHA; SEQ ID NO:28) or two other scrambled sequences (#3: YRCETDSPHEFHLWNTMCAF; SEQ ID NO:29 and #4: YRCETDSPHEFHLWNTFCAM; SEQ ID NO:30) also showed no tumor targeting activity for the mutants.
- CRKL-binding phage was at least 10- fold higher when compared to an ⁇ v integrin-binding phage (peptide sequence CDCRGDCFC (SEQ ID NO:31), termed RGD-4C) which is typically used as a positive control for this type of experiment (Hajitou et al, 2006; Arap et al, 1998; Pasqualini et al, 2001; Giordano et al, 2001; Javadpour et al, 1996; Ellerby et al, 1999; Pasqualini et al, 1997).
- Animals used inexperiments were: Male nude mice bearing human DU145 xenografts or female nude mice with human Kaposi sarcoma KS1767-derived xenografts subcutaneously and immunocompetent Balb/c female mice bearing EF43-FGF4-derived breast tumors orthotopically in the mammary fat pad.
- mice bearing tumors were anesthetized and injected intravenously via tail vein with 5 xlO 10 T.U. per mouse of wild-type YRCTLNSPFFWEDMTHECHA (SEQ ID NO:l)-phage, or negative controls: fd- tet phage (insertless) and scrambled YRFCTSPFHEWHLENTDMCA (SEQ ID NO:26)- phage, YRECTDSPHEFHLWNTMCAF (SEQ ID NO:27)-phage,
- YRCETDSPHEFHLWNTMCAF SEQ ID NO:29
- YRCETDSPHEFHLWNTFCAM SEQ ID NO:30
- mutants YRCTLNSAFFWEDMTHECHA SEQ ID NO:25
- the CRKL homing phage was first incubated with the recombinant gst-CRKL or control gst protein for 30 minutes at 37 0 C, then intravenously administered into prostate tumor-bearing mice.
- the fd phage served a negative control and the RGD-4C (Hajitou et ⁇ l., 2006; Arap et ⁇ l., 1998) served as a positive control.
- Only minimal background apoptosis ( ⁇ 1% of the total cells) were detected by TUNEL staining (Promega) on paraffin-embedded tumor tissue sections. Therapy in tumor-bearing mice
- the tumor-homing peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO: 1) was synthesized fused with the proapoptotic motif D (KL AKL AK) 2 .
- Unconjugated peptide YRCTLNSPFFWEDMTHECHA (SEQ ID NO:1) or D (KLAKLAK) 2 served as controls.
- the synthetic peptides were systemically administered and tumor volumes measured as described (Arap et ⁇ l, 1998; Arap et ⁇ l, 2004).
- Gold/Phage imidazole hydrogels were formed with tumor-targeting phage (targeting CRKL) and insertless phage as a control. Iron-oxide was incorporated into these hydrogels at a final volume of 30% (by volume).
- This hydrogel preparation was used in a subsequent MRI-study in which three prostate tumor-bearing mice (DU145) were injected intra-tumorally with equivalent amounts of AuFe only, untargeted hydrogels (with insertless phage) containing AuFe and CRKL-targeted hydrogels containing AuFe.
- the negative contrast mediated by the iron-core can clearly be seen and quantified using MRI.
- Nicolas and Rubinstein In: Vectors: A survey of molecular cloning vectors and their uses, Rodriguez and Denhardt, eds., Stoneham: Butterworth, pp. 494-513, 1988.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Toxicology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Oncology (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
Claims
Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/000,231 US20110189095A1 (en) | 2008-06-20 | 2009-06-19 | Crkl targeting peptides |
CN2009801292540A CN102105487A (en) | 2008-06-20 | 2009-06-19 | CRKL targeting peptides |
JP2011514861A JP2011525491A (en) | 2008-06-20 | 2009-06-19 | CRKL targeting peptide |
EP09767865.0A EP2303913A4 (en) | 2008-06-20 | 2009-06-19 | Crkl targeting peptides |
CA2728763A CA2728763A1 (en) | 2008-06-20 | 2009-06-19 | Crkl targeting peptides |
MX2010014173A MX2010014173A (en) | 2008-06-20 | 2009-06-19 | Crkl targeting peptides. |
BRPI0915718A BRPI0915718A2 (en) | 2008-06-20 | 2009-06-19 | crkl steering peptides |
IL210053A IL210053A0 (en) | 2008-06-20 | 2010-12-16 | Crkl targeting peptides |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US7442308P | 2008-06-20 | 2008-06-20 | |
US61/074,423 | 2008-06-20 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2009155556A2 true WO2009155556A2 (en) | 2009-12-23 |
WO2009155556A3 WO2009155556A3 (en) | 2010-04-08 |
Family
ID=41434721
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2009/048024 WO2009155556A2 (en) | 2008-06-20 | 2009-06-19 | Crkl targeting peptides |
Country Status (14)
Country | Link |
---|---|
US (1) | US20110189095A1 (en) |
EP (1) | EP2303913A4 (en) |
JP (1) | JP2011525491A (en) |
CN (1) | CN102105487A (en) |
BR (1) | BRPI0915718A2 (en) |
CA (1) | CA2728763A1 (en) |
CL (1) | CL2010001498A1 (en) |
CO (1) | CO6331291A2 (en) |
CR (1) | CR20110034A (en) |
EC (1) | ECSP10010729A (en) |
IL (1) | IL210053A0 (en) |
MX (1) | MX2010014173A (en) |
PE (1) | PE20110309A1 (en) |
WO (1) | WO2009155556A2 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011147057A1 (en) * | 2010-05-25 | 2011-12-01 | 北京大学 | Anti-angiogenic fusion polypeptide, gene and use thereof |
JP2014513066A (en) * | 2011-03-21 | 2014-05-29 | セントロ デ インジエニエリア ジエネテイカ イ バイオテクノロジア | Cyclic peptides having anti-neoplastic and anti-angiogenic activity |
WO2015035881A1 (en) * | 2013-09-11 | 2015-03-19 | 中山大学附属肿瘤医院 | Tumor-targeting polypeptide and application thereof |
CN110025577A (en) * | 2019-03-19 | 2019-07-19 | 广东药科大学 | A kind of polypeptide drugs take orally targeted system M27-39@FA-MCNs complex and its preparation method and application |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP6505691B2 (en) * | 2013-07-25 | 2019-04-24 | ノバルティス アーゲー | Cyclic Apelin Derivatives for the Treatment of Heart Failure |
US9340582B2 (en) * | 2013-07-25 | 2016-05-17 | Novartis Ag | Bioconjugates of synthetic apelin polypeptides |
WO2015013167A1 (en) * | 2013-07-25 | 2015-01-29 | Novartis Ag | Disulfide cyclic polypeptides for the treatment of heart failure |
CN105612172A (en) * | 2013-07-25 | 2016-05-25 | 诺华股份有限公司 | Cyclic polypeptides for the treatment of heart failure |
AU2015209007A1 (en) | 2014-01-27 | 2016-09-15 | The Children's Hospital Of Philadelphia | Compositions and methods for treating autoimmune and inflammatory diseases |
CN108303539B (en) * | 2018-01-31 | 2020-08-11 | 刘双萍 | Biological reagent for detecting breast cancer cells and application thereof |
CN109908369B (en) * | 2019-04-28 | 2022-02-11 | 复旦大学附属金山医院 | Application of novel circular RNA circCRKL in prostate cancer treatment |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5667981A (en) * | 1994-05-13 | 1997-09-16 | Childrens Hospital Of Los Angeles | Diagnostics and treatments for cancers expressing tyrosine phosphorylated CRKL protein |
US7309692B1 (en) * | 1996-07-08 | 2007-12-18 | Board Of Regents, The University Of Texas System | Inhibition of chronic myelogenous leukemic cell growth by liposomal-antisense oligodeoxy-nucleotides targeting to GRB2 or CRK1 |
WO2004020999A1 (en) * | 2002-08-30 | 2004-03-11 | Board Of Regents, The University Of Texas System | Compositions and methods of use of targeting peptides for diagnosis and therapy of human cancer |
US7671010B2 (en) * | 2002-08-30 | 2010-03-02 | The Board Of Regents Of The University Of Texas System | Compositions and methods of use of targeting peptides for diagnosis and therapy of human cancer |
AU2005307789A1 (en) * | 2004-11-16 | 2006-05-26 | Avidia Research Institute | Protein scaffolds and uses thereof |
US20120017338A1 (en) * | 2008-01-15 | 2012-01-19 | Wei Wu | Isolated novel nucleic acid and protein molecules from corn and methods of using those molecules to generate transgenic plant with enhanced agronomic traits |
-
2009
- 2009-06-19 BR BRPI0915718A patent/BRPI0915718A2/en not_active IP Right Cessation
- 2009-06-19 CN CN2009801292540A patent/CN102105487A/en active Pending
- 2009-06-19 CA CA2728763A patent/CA2728763A1/en not_active Abandoned
- 2009-06-19 MX MX2010014173A patent/MX2010014173A/en not_active Application Discontinuation
- 2009-06-19 JP JP2011514861A patent/JP2011525491A/en not_active Withdrawn
- 2009-06-19 US US13/000,231 patent/US20110189095A1/en not_active Abandoned
- 2009-06-19 WO PCT/US2009/048024 patent/WO2009155556A2/en active Application Filing
- 2009-06-19 PE PE2010001154A patent/PE20110309A1/en not_active Application Discontinuation
- 2009-06-19 EP EP09767865.0A patent/EP2303913A4/en not_active Withdrawn
-
2010
- 2010-12-16 IL IL210053A patent/IL210053A0/en unknown
- 2010-12-20 CO CO10159776A patent/CO6331291A2/en not_active Application Discontinuation
- 2010-12-20 CL CL2010001498A patent/CL2010001498A1/en unknown
- 2010-12-29 EC EC2010010729A patent/ECSP10010729A/en unknown
-
2011
- 2011-01-20 CR CR20110034A patent/CR20110034A/en unknown
Non-Patent Citations (1)
Title |
---|
See references of EP2303913A4 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011147057A1 (en) * | 2010-05-25 | 2011-12-01 | 北京大学 | Anti-angiogenic fusion polypeptide, gene and use thereof |
JP2014513066A (en) * | 2011-03-21 | 2014-05-29 | セントロ デ インジエニエリア ジエネテイカ イ バイオテクノロジア | Cyclic peptides having anti-neoplastic and anti-angiogenic activity |
WO2015035881A1 (en) * | 2013-09-11 | 2015-03-19 | 中山大学附属肿瘤医院 | Tumor-targeting polypeptide and application thereof |
US9809622B2 (en) | 2013-09-11 | 2017-11-07 | Sun Yat-Sen University Cancer Center | Tumor-targeting polypeptide and application thereof |
CN110025577A (en) * | 2019-03-19 | 2019-07-19 | 广东药科大学 | A kind of polypeptide drugs take orally targeted system M27-39@FA-MCNs complex and its preparation method and application |
CN110025577B (en) * | 2019-03-19 | 2021-09-07 | 广东药科大学 | Polypeptide drug oral targeting system M27-39@ FA-MCNs complex and preparation method and application thereof |
Also Published As
Publication number | Publication date |
---|---|
ECSP10010729A (en) | 2011-04-29 |
BRPI0915718A2 (en) | 2017-06-20 |
WO2009155556A3 (en) | 2010-04-08 |
CL2010001498A1 (en) | 2011-05-20 |
CO6331291A2 (en) | 2011-10-20 |
US20110189095A1 (en) | 2011-08-04 |
EP2303913A4 (en) | 2013-07-24 |
PE20110309A1 (en) | 2011-06-19 |
EP2303913A2 (en) | 2011-04-06 |
JP2011525491A (en) | 2011-09-22 |
IL210053A0 (en) | 2011-02-28 |
CR20110034A (en) | 2011-06-24 |
CA2728763A1 (en) | 2009-12-23 |
CN102105487A (en) | 2011-06-22 |
MX2010014173A (en) | 2011-06-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20110189095A1 (en) | Crkl targeting peptides | |
CUSSAC et al. | A Sos‐derived peptidimer blocks the Ras signaling pathway by binding both Grb2 SH3 domains and displays antiproliferative activity | |
AU2008296733B2 (en) | VEGFR-1/NRP-1 targeting peptides | |
CN101146822A (en) | EphB receptor-binding peptides | |
CN110092817A (en) | Cell-penetrating peptides, the conjugate comprising the peptide and the composition comprising the conjugate | |
US7914780B1 (en) | Aminopeptidase A (APA) targeting peptides for the treatment of cancer | |
US9527895B2 (en) | CAPCNA peptide therapeutics for cancer | |
JP2012065662A (en) | Human and mouse targeting peptide identified by phage display | |
WO2006010070A2 (en) | Compositions and methods related to peptides that selectively bind leukemia cells | |
JP2009502983A (en) | Protein kinase C isoform inhibitors and uses thereof | |
JP2009527498A (en) | Inhibition of caPCNA interaction in cancer by peptides | |
JP2011525491A5 (en) | ||
KR20120125455A (en) | Intracelluar targeting bipodal peptide binder | |
US20130296252A1 (en) | Muc18 targeting peptides | |
JP5077862B2 (en) | Human and mouse targeting peptides identified by phage display | |
JP2004508045A5 (en) | ||
US11407788B2 (en) | Prostate-specific membrane antigen (PSMA) targeting peptides | |
Bussolati et al. | Targeting of human renal tumor-derived endothelial cells with peptides obtained by phage display | |
US20130059793A1 (en) | Egf receptor mimicking peptides | |
KR101323846B1 (en) | D-Aptide Having Maintained Target Affinity and Enhanced Stability | |
CA2367256A1 (en) | Surface localized colligin/hsp47 in carcinoma cells | |
CN105859841B (en) | A kind of pair targets chimeric peptide and its is preparing the application in medicine for anti transfer of tumor | |
US20220096594A1 (en) | Macrocyclic peptides for targeted inhibition of autophagy | |
WO2013149237A1 (en) | Targeting intracellular organelle zip codes with functional homing ligands from cell-internalizing phage combinatorial libraries | |
US20140356285A1 (en) | Compositions and methods related to tissue targeting |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 200980129254.0 Country of ref document: CN |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 09767865 Country of ref document: EP Kind code of ref document: A2 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 210053 Country of ref document: IL Ref document number: 001154-2010 Country of ref document: PE |
|
ENP | Entry into the national phase |
Ref document number: 2011514861 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: MX/A/2010/014173 Country of ref document: MX |
|
ENP | Entry into the national phase |
Ref document number: 2728763 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2010001498 Country of ref document: CL Ref document number: 10159776 Country of ref document: CO |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 446/DELNP/2011 Country of ref document: IN |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2009767865 Country of ref document: EP Ref document number: CR2011-000034 Country of ref document: CR |
|
WWE | Wipo information: entry into national phase |
Ref document number: 13000231 Country of ref document: US |
|
ENP | Entry into the national phase |
Ref document number: PI0915718 Country of ref document: BR Kind code of ref document: A2 Effective date: 20101220 |