US20210023116A1 - Treatment of cancer by systemic administration of dbait molecules - Google Patents
Treatment of cancer by systemic administration of dbait molecules Download PDFInfo
- Publication number
- US20210023116A1 US20210023116A1 US17/038,704 US202017038704A US2021023116A1 US 20210023116 A1 US20210023116 A1 US 20210023116A1 US 202017038704 A US202017038704 A US 202017038704A US 2021023116 A1 US2021023116 A1 US 2021023116A1
- Authority
- US
- United States
- Prior art keywords
- nucleic acid
- cancer
- treatment
- group
- dna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 238000011282 treatment Methods 0.000 title claims description 117
- 206010028980 Neoplasm Diseases 0.000 title description 130
- 201000011510 cancer Diseases 0.000 title description 48
- 238000007910 systemic administration Methods 0.000 title description 6
- 230000009885 systemic effect Effects 0.000 claims abstract description 20
- 230000000021 endosomolytic effect Effects 0.000 claims abstract description 18
- 108020004707 nucleic acids Proteins 0.000 claims description 63
- 102000039446 nucleic acids Human genes 0.000 claims description 63
- 150000007523 nucleic acids Chemical class 0.000 claims description 62
- 238000002512 chemotherapy Methods 0.000 claims description 49
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 46
- 125000003729 nucleotide group Chemical group 0.000 claims description 43
- 239000002773 nucleotide Substances 0.000 claims description 42
- 239000002246 antineoplastic agent Substances 0.000 claims description 37
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 claims description 33
- 229960001756 oxaliplatin Drugs 0.000 claims description 33
- 238000001990 intravenous administration Methods 0.000 claims description 32
- 238000007912 intraperitoneal administration Methods 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 27
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 claims description 24
- 235000012000 cholesterol Nutrition 0.000 claims description 23
- 108020004414 DNA Proteins 0.000 claims description 22
- 229960004562 carboplatin Drugs 0.000 claims description 22
- 239000003795 chemical substances by application Substances 0.000 claims description 22
- 229940107161 cholesterol Drugs 0.000 claims description 21
- 238000001959 radiotherapy Methods 0.000 claims description 20
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 18
- 229920001223 polyethylene glycol Polymers 0.000 claims description 16
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 14
- 238000002347 injection Methods 0.000 claims description 14
- 150000004713 phosphodiesters Chemical class 0.000 claims description 14
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 12
- 239000003112 inhibitor Substances 0.000 claims description 12
- 206010033128 Ovarian cancer Diseases 0.000 claims description 11
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 11
- 230000012202 endocytosis Effects 0.000 claims description 11
- IIRDTKBZINWQAW-UHFFFAOYSA-N hexaethylene glycol Chemical compound OCCOCCOCCOCCOCCOCCO IIRDTKBZINWQAW-UHFFFAOYSA-N 0.000 claims description 11
- 108090000623 proteins and genes Proteins 0.000 claims description 11
- 229910052697 platinum Inorganic materials 0.000 claims description 9
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical group OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 claims description 8
- 230000000259 anti-tumor effect Effects 0.000 claims description 8
- 125000005518 carboxamido group Chemical group 0.000 claims description 8
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims description 8
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 8
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 claims description 8
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 claims description 8
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims description 8
- 229960001295 tocopherol Drugs 0.000 claims description 8
- 229930003799 tocopherol Natural products 0.000 claims description 8
- 235000010384 tocopherol Nutrition 0.000 claims description 8
- 239000011732 tocopherol Substances 0.000 claims description 8
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 7
- 239000003446 ligand Substances 0.000 claims description 7
- YACKEPLHDIMKIO-UHFFFAOYSA-N methylphosphonic acid Chemical group CP(O)(O)=O YACKEPLHDIMKIO-UHFFFAOYSA-N 0.000 claims description 7
- 108020003175 receptors Proteins 0.000 claims description 7
- 102000005962 receptors Human genes 0.000 claims description 7
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 6
- UWHCKJMYHZGTIT-UHFFFAOYSA-N Tetraethylene glycol, Natural products OCCOCCOCCOCCO UWHCKJMYHZGTIT-UHFFFAOYSA-N 0.000 claims description 6
- 229960000304 folic acid Drugs 0.000 claims description 6
- 235000019152 folic acid Nutrition 0.000 claims description 6
- 239000011724 folic acid Substances 0.000 claims description 6
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 6
- 239000004971 Cross linker Substances 0.000 claims description 5
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims description 5
- 101710183280 Topoisomerase Proteins 0.000 claims description 5
- 239000002256 antimetabolite Substances 0.000 claims description 5
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 5
- 229960004316 cisplatin Drugs 0.000 claims description 5
- 229930182830 galactose Natural products 0.000 claims description 5
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 4
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 4
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 4
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 4
- 229930024421 Adenine Natural products 0.000 claims description 4
- 108010051479 Bombesin Proteins 0.000 claims description 4
- 102000013585 Bombesin Human genes 0.000 claims description 4
- 108091029430 CpG site Proteins 0.000 claims description 4
- 239000005642 Oleic acid Substances 0.000 claims description 4
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000002202 Polyethylene glycol Substances 0.000 claims description 4
- 235000021355 Stearic acid Nutrition 0.000 claims description 4
- 229960000643 adenine Drugs 0.000 claims description 4
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 claims description 4
- DNDCVAGJPBKION-DOPDSADYSA-N bombesin Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1NC2=CC=CC=C2C=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1NC(=O)CC1)C(C)C)C1=CN=CN1 DNDCVAGJPBKION-DOPDSADYSA-N 0.000 claims description 4
- 229940104302 cytosine Drugs 0.000 claims description 4
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 4
- 229930195729 fatty acid Natural products 0.000 claims description 4
- 239000000194 fatty acid Substances 0.000 claims description 4
- 150000004665 fatty acids Chemical class 0.000 claims description 4
- 102000006495 integrins Human genes 0.000 claims description 4
- 108010044426 integrins Proteins 0.000 claims description 4
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 4
- 230000000394 mitotic effect Effects 0.000 claims description 4
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 4
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 4
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 4
- 229920001542 oligosaccharide Polymers 0.000 claims description 4
- 150000002482 oligosaccharides Chemical class 0.000 claims description 4
- 239000008117 stearic acid Substances 0.000 claims description 4
- 229940113082 thymine Drugs 0.000 claims description 4
- 229940126161 DNA alkylating agent Drugs 0.000 claims description 3
- 239000012624 DNA alkylating agent Substances 0.000 claims description 3
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 3
- 150000002634 lipophilic molecules Chemical class 0.000 claims description 3
- 230000010837 receptor-mediated endocytosis Effects 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 229940079593 drug Drugs 0.000 claims description 2
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 claims 1
- 102000004338 Transferrin Human genes 0.000 claims 1
- 108090000901 Transferrin Proteins 0.000 claims 1
- 190000008236 carboplatin Chemical compound 0.000 claims 1
- 239000012581 transferrin Substances 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 58
- 210000004185 liver Anatomy 0.000 description 32
- 206010009944 Colon cancer Diseases 0.000 description 29
- 208000003721 Triple Negative Breast Neoplasms Diseases 0.000 description 29
- 239000008194 pharmaceutical composition Substances 0.000 description 29
- 208000022679 triple-negative breast carcinoma Diseases 0.000 description 29
- 125000005647 linker group Chemical group 0.000 description 28
- 229960002949 fluorouracil Drugs 0.000 description 27
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 26
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 25
- 208000032839 leukemia Diseases 0.000 description 24
- 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 23
- 206010027476 Metastases Diseases 0.000 description 18
- 241000699670 Mus sp. Species 0.000 description 18
- 241001465754 Metazoa Species 0.000 description 17
- 230000009401 metastasis Effects 0.000 description 16
- 238000002360 preparation method Methods 0.000 description 16
- 230000000694 effects Effects 0.000 description 15
- 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 14
- 208000014018 liver neoplasm Diseases 0.000 description 14
- 230000001394 metastastic effect Effects 0.000 description 12
- 206010061289 metastatic neoplasm Diseases 0.000 description 12
- 230000004083 survival effect Effects 0.000 description 12
- -1 RGD and bombesin Chemical class 0.000 description 11
- 230000001225 therapeutic effect Effects 0.000 description 11
- 230000035755 proliferation Effects 0.000 description 10
- 210000001519 tissue Anatomy 0.000 description 10
- 206010006187 Breast cancer Diseases 0.000 description 9
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 9
- 239000004480 active ingredient Substances 0.000 description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 108090000765 processed proteins & peptides Proteins 0.000 description 9
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 8
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 8
- 208000026310 Breast neoplasm Diseases 0.000 description 8
- 201000001441 melanoma Diseases 0.000 description 8
- 230000035945 sensitivity Effects 0.000 description 8
- 230000004614 tumor growth Effects 0.000 description 8
- WHTVZRBIWZFKQO-AWEZNQCLSA-N (S)-chloroquine Chemical compound ClC1=CC=C2C(N[C@@H](C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-AWEZNQCLSA-N 0.000 description 7
- 230000033616 DNA repair Effects 0.000 description 7
- 206010019695 Hepatic neoplasm Diseases 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 229960003677 chloroquine Drugs 0.000 description 7
- WHTVZRBIWZFKQO-UHFFFAOYSA-N chloroquine Natural products ClC1=CC=C2C(NC(C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-UHFFFAOYSA-N 0.000 description 7
- 230000007423 decrease Effects 0.000 description 7
- 201000010099 disease Diseases 0.000 description 7
- 229960004679 doxorubicin Drugs 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 238000000338 in vitro Methods 0.000 description 7
- 238000001727 in vivo Methods 0.000 description 7
- 231100000419 toxicity Toxicity 0.000 description 7
- 230000001988 toxicity Effects 0.000 description 7
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 6
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 description 6
- 108010006124 DNA-Activated Protein Kinase Proteins 0.000 description 6
- 102000005768 DNA-Activated Protein Kinase Human genes 0.000 description 6
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 description 6
- 206010029260 Neuroblastoma Diseases 0.000 description 6
- 230000006907 apoptotic process Effects 0.000 description 6
- 238000011254 conventional chemotherapy Methods 0.000 description 6
- 239000003937 drug carrier Substances 0.000 description 6
- 230000002601 intratumoral effect Effects 0.000 description 6
- 201000007270 liver cancer Diseases 0.000 description 6
- 208000008585 mastocytosis Diseases 0.000 description 6
- 230000017074 necrotic cell death Effects 0.000 description 6
- 238000013421 nuclear magnetic resonance imaging Methods 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- MZKSKFARYTYMLV-UHFFFAOYSA-N CCN.CCN.[N-]=[N+]=NCN Chemical compound CCN.CCN.[N-]=[N+]=NCN MZKSKFARYTYMLV-UHFFFAOYSA-N 0.000 description 5
- 208000032612 Glial tumor Diseases 0.000 description 5
- 206010018338 Glioma Diseases 0.000 description 5
- 206010025323 Lymphomas Diseases 0.000 description 5
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 5
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 5
- 230000004913 activation Effects 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 239000007928 intraperitoneal injection Substances 0.000 description 5
- 210000003734 kidney Anatomy 0.000 description 5
- 229960001592 paclitaxel Drugs 0.000 description 5
- 210000004303 peritoneum Anatomy 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 208000036762 Acute promyelocytic leukaemia Diseases 0.000 description 4
- 206010005003 Bladder cancer Diseases 0.000 description 4
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 description 4
- 102100034533 Histone H2AX Human genes 0.000 description 4
- 208000008839 Kidney Neoplasms Diseases 0.000 description 4
- 108060001084 Luciferase Proteins 0.000 description 4
- 239000005089 Luciferase Substances 0.000 description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 4
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 4
- 206010030113 Oedema Diseases 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 206010038389 Renal cancer Diseases 0.000 description 4
- 206010039491 Sarcoma Diseases 0.000 description 4
- 206010041067 Small cell lung cancer Diseases 0.000 description 4
- 208000005718 Stomach Neoplasms Diseases 0.000 description 4
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 206010017758 gastric cancer Diseases 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 201000010536 head and neck cancer Diseases 0.000 description 4
- 208000014829 head and neck neoplasm Diseases 0.000 description 4
- 201000005787 hematologic cancer Diseases 0.000 description 4
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 4
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 4
- 238000003364 immunohistochemistry Methods 0.000 description 4
- 201000010982 kidney cancer Diseases 0.000 description 4
- 201000005202 lung cancer Diseases 0.000 description 4
- 208000020816 lung neoplasm Diseases 0.000 description 4
- 208000003747 lymphoid leukemia Diseases 0.000 description 4
- 230000003211 malignant effect Effects 0.000 description 4
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- BWKDAMBGCPRVPI-ZQRPHVBESA-N ortataxel Chemical compound O([C@@H]1[C@]23OC(=O)O[C@H]2[C@@H](C(=C([C@@H](OC(C)=O)C(=O)[C@]2(C)[C@@H](O)C[C@H]4OC[C@]4([C@H]21)OC(C)=O)C3(C)C)C)OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)CC(C)C)C(=O)C1=CC=CC=C1 BWKDAMBGCPRVPI-ZQRPHVBESA-N 0.000 description 4
- 230000026731 phosphorylation Effects 0.000 description 4
- 238000006366 phosphorylation reaction Methods 0.000 description 4
- 208000000587 small cell lung carcinoma Diseases 0.000 description 4
- 206010041823 squamous cell carcinoma Diseases 0.000 description 4
- 201000011549 stomach cancer Diseases 0.000 description 4
- 238000007920 subcutaneous administration Methods 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 210000001685 thyroid gland Anatomy 0.000 description 4
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 4
- 201000005112 urinary bladder cancer Diseases 0.000 description 4
- 108091023037 Aptamer Proteins 0.000 description 3
- 201000009030 Carcinoma Diseases 0.000 description 3
- 229960005500 DHA-paclitaxel Drugs 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 3
- 239000012623 DNA damaging agent Substances 0.000 description 3
- 102100022204 DNA-dependent protein kinase catalytic subunit Human genes 0.000 description 3
- 101710157074 DNA-dependent protein kinase catalytic subunit Proteins 0.000 description 3
- 201000008808 Fibrosarcoma Diseases 0.000 description 3
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 3
- 101001067891 Homo sapiens Histone H2AX Proteins 0.000 description 3
- 102000007330 LDL Lipoproteins Human genes 0.000 description 3
- 108010007622 LDL Lipoproteins Proteins 0.000 description 3
- 208000028018 Lymphocytic leukaemia Diseases 0.000 description 3
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 3
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 3
- 201000010208 Seminoma Diseases 0.000 description 3
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 3
- 230000001154 acute effect Effects 0.000 description 3
- 210000000577 adipose tissue Anatomy 0.000 description 3
- 230000033115 angiogenesis Effects 0.000 description 3
- 201000010881 cervical cancer Diseases 0.000 description 3
- 230000003034 chemosensitisation Effects 0.000 description 3
- 208000029742 colonic neoplasm Diseases 0.000 description 3
- 201000010989 colorectal carcinoma Diseases 0.000 description 3
- 238000011284 combination treatment Methods 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- LRCZQSDQZJBHAF-PUBGEWHCSA-N dha-paclitaxel Chemical compound N([C@H]([C@@H](OC(=O)CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CC)C(=O)O[C@@H]1C(=C2[C@@H](OC(C)=O)C(=O)[C@]3(C)[C@@H](O)C[C@H]4OC[C@]4([C@H]3[C@H](OC(=O)C=3C=CC=CC=3)[C@](C2(C)C)(O)C1)OC(C)=O)C)C=1C=CC=CC=1)C(=O)C1=CC=CC=C1 LRCZQSDQZJBHAF-PUBGEWHCSA-N 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 208000005017 glioblastoma Diseases 0.000 description 3
- 210000000936 intestine Anatomy 0.000 description 3
- 208000020442 loss of weight Diseases 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 210000004088 microvessel Anatomy 0.000 description 3
- 208000025113 myeloid leukemia Diseases 0.000 description 3
- 230000001338 necrotic effect Effects 0.000 description 3
- 231100000956 nontoxicity Toxicity 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 238000010837 poor prognosis Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 201000009410 rhabdomyosarcoma Diseases 0.000 description 3
- 231100000161 signs of toxicity Toxicity 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 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 description 3
- 208000001608 teratocarcinoma Diseases 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 239000003981 vehicle Substances 0.000 description 3
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 2
- 108010082126 Alanine transaminase Proteins 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 2
- 102000013142 Amylases Human genes 0.000 description 2
- 108010065511 Amylases Proteins 0.000 description 2
- 206010003571 Astrocytoma Diseases 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 2
- 208000003950 B-cell lymphoma Diseases 0.000 description 2
- 208000018084 Bone neoplasm Diseases 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N CCC Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- LRRLHMZLZKHQHP-OHGAXJPQSA-N CCCOCCOCCOCCCCC(=O)O[C@H]1CC[C@@]2(C)C(=CCC3C2CC[C@@]2(C)C3CCC2[C@H](C)CCCC(C)C)C1 Chemical compound CCCOCCOCCOCCCCC(=O)O[C@H]1CC[C@@]2(C)C(=CCC3C2CC[C@@]2(C)C3CCC2[C@H](C)CCCC(C)C)C1 LRRLHMZLZKHQHP-OHGAXJPQSA-N 0.000 description 2
- 206010008342 Cervix carcinoma Diseases 0.000 description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 2
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 206010014733 Endometrial cancer Diseases 0.000 description 2
- 206010014759 Endometrial neoplasm Diseases 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 208000021309 Germ cell tumor Diseases 0.000 description 2
- 201000010915 Glioblastoma multiforme Diseases 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 208000017604 Hodgkin disease Diseases 0.000 description 2
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 2
- 208000010747 Hodgkins lymphoma Diseases 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 208000007766 Kaposi sarcoma Diseases 0.000 description 2
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 206010027457 Metastases to liver Diseases 0.000 description 2
- 241000204031 Mycoplasma Species 0.000 description 2
- 208000034176 Neoplasms, Germ Cell and Embryonal Diseases 0.000 description 2
- WVGDHYDGENAOQB-KPNWGBFJSA-N OCCOCCOCCOCCO.C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 Chemical compound OCCOCCOCCOCCO.C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 WVGDHYDGENAOQB-KPNWGBFJSA-N 0.000 description 2
- 229930012538 Paclitaxel Natural products 0.000 description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 2
- 108091093037 Peptide nucleic acid Proteins 0.000 description 2
- 239000004952 Polyamide Substances 0.000 description 2
- 229920002873 Polyethylenimine Polymers 0.000 description 2
- 239000004721 Polyphenylene oxide Substances 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 206010042971 T-cell lymphoma Diseases 0.000 description 2
- 208000027585 T-cell non-Hodgkin lymphoma Diseases 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 208000024770 Thyroid neoplasm Diseases 0.000 description 2
- 238000008050 Total Bilirubin Reagent Methods 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 208000036676 acute undifferentiated leukemia Diseases 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 235000019418 amylase Nutrition 0.000 description 2
- 229940045799 anthracyclines and related substance Drugs 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 230000004700 cellular uptake Effects 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 210000003679 cervix uteri Anatomy 0.000 description 2
- 125000003636 chemical group Chemical group 0.000 description 2
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 229940127089 cytotoxic agent Drugs 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 230000002939 deleterious effect Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000003325 follicular Effects 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 201000011243 gastrointestinal stromal tumor Diseases 0.000 description 2
- 150000002334 glycols Chemical class 0.000 description 2
- 230000002440 hepatic effect Effects 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- SEFGUGYLLVNFIJ-QDRLFVHASA-N larotaxel dihydrate Chemical compound O.O.O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@@]23[C@H]1[C@@]1(CO[C@@H]1C[C@@H]2C3)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 SEFGUGYLLVNFIJ-QDRLFVHASA-N 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 230000036210 malignancy Effects 0.000 description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 2
- 208000000516 mast-cell leukemia Diseases 0.000 description 2
- GLVAUDGFNGKCSF-UHFFFAOYSA-N mercaptopurine Chemical compound S=C1NC=NC2=C1NC=N2 GLVAUDGFNGKCSF-UHFFFAOYSA-N 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 2
- 201000006894 monocytic leukemia Diseases 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 208000007538 neurilemmoma Diseases 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 201000008968 osteosarcoma Diseases 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- 108700027936 paclitaxel poliglumex Proteins 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 201000002528 pancreatic cancer Diseases 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- 230000007170 pathology Effects 0.000 description 2
- 210000001428 peripheral nervous system Anatomy 0.000 description 2
- 210000004214 philadelphia chromosome Anatomy 0.000 description 2
- 208000031223 plasma cell leukemia Diseases 0.000 description 2
- 229920002647 polyamide Polymers 0.000 description 2
- 229920000768 polyamine Polymers 0.000 description 2
- 229920000570 polyether Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 208000016691 refractory malignant neoplasm Diseases 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 239000004248 saffron Substances 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- MODVSQKJJIBWPZ-VLLPJHQWSA-N tesetaxel Chemical compound O([C@H]1[C@@H]2[C@]3(OC(C)=O)CO[C@@H]3CC[C@@]2(C)[C@H]2[C@@H](C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C(=CC=CN=4)F)C[C@]1(O)C3(C)C)O[C@H](O2)CN(C)C)C(=O)C1=CC=CC=C1 MODVSQKJJIBWPZ-VLLPJHQWSA-N 0.000 description 2
- 210000001550 testis Anatomy 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 201000002510 thyroid cancer Diseases 0.000 description 2
- WYWHKKSPHMUBEB-UHFFFAOYSA-N tioguanine Chemical compound N1C(N)=NC(=S)C2=C1N=CN2 WYWHKKSPHMUBEB-UHFFFAOYSA-N 0.000 description 2
- 230000001173 tumoral effect Effects 0.000 description 2
- 210000003932 urinary bladder Anatomy 0.000 description 2
- ZPUHVPYXSITYDI-HEUWMMRCSA-N xyotax Chemical compound OC(=O)[C@@H](N)CCC(O)=O.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 ZPUHVPYXSITYDI-HEUWMMRCSA-N 0.000 description 2
- YXTKHLHCVFUPPT-YYFJYKOTSA-N (2s)-2-[[4-[(2-amino-5-formyl-4-oxo-1,6,7,8-tetrahydropteridin-6-yl)methylamino]benzoyl]amino]pentanedioic acid;(1r,2r)-1,2-dimethanidylcyclohexane;5-fluoro-1h-pyrimidine-2,4-dione;oxalic acid;platinum(2+) Chemical compound [Pt+2].OC(=O)C(O)=O.[CH2-][C@@H]1CCCC[C@H]1[CH2-].FC1=CNC(=O)NC1=O.C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 YXTKHLHCVFUPPT-YYFJYKOTSA-N 0.000 description 1
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 1
- 0 *CCCCCCCCCC(NCCCOCC(*)O)=O Chemical compound *CCCCCCCCCC(NCCCOCC(*)O)=O 0.000 description 1
- 108010058566 130-nm albumin-bound paclitaxel Proteins 0.000 description 1
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-VTZDEGQISA-N 0.000 description 1
- IDPUKCWIGUEADI-UHFFFAOYSA-N 5-[bis(2-chloroethyl)amino]uracil Chemical compound ClCCN(CCCl)C1=CNC(=O)NC1=O IDPUKCWIGUEADI-UHFFFAOYSA-N 0.000 description 1
- 208000035657 Abasia Diseases 0.000 description 1
- 206010000830 Acute leukaemia Diseases 0.000 description 1
- 206010000871 Acute monocytic leukaemia Diseases 0.000 description 1
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 1
- 208000009746 Adult T-Cell Leukemia-Lymphoma Diseases 0.000 description 1
- 208000016683 Adult T-cell leukemia/lymphoma Diseases 0.000 description 1
- 208000035805 Aleukaemic leukaemia Diseases 0.000 description 1
- 102000007325 Amelogenin Human genes 0.000 description 1
- 108010007570 Amelogenin Proteins 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 1
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 1
- NOWKCMXCCJGMRR-UHFFFAOYSA-N Aziridine Chemical class C1CN1 NOWKCMXCCJGMRR-UHFFFAOYSA-N 0.000 description 1
- 102000036365 BRCA1 Human genes 0.000 description 1
- 108700020463 BRCA1 Proteins 0.000 description 1
- 101150072950 BRCA1 gene Proteins 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 206010005949 Bone cancer Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- CUTQGAGLIQSTDG-UHFFFAOYSA-N C.C.C.C.CCC.CCC.CCC.CCC.CCC Chemical compound C.C.C.C.CCC.CCC.CCC.CCC.CCC CUTQGAGLIQSTDG-UHFFFAOYSA-N 0.000 description 1
- ZBZOKCYEHDGDPV-UHFFFAOYSA-N C.C.C.CCC.CCC.CCC.CCC.CCC.CCN.CCN.CCN.CCN.CCN Chemical compound C.C.C.CCC.CCC.CCC.CCC.CCC.CCN.CCN.CCN.CCN.CCN ZBZOKCYEHDGDPV-UHFFFAOYSA-N 0.000 description 1
- VEMFGJIHJBILSW-ZVVYFNDBSA-N C.COCCCCCCCCCC(=O)CCCCOCC(CO)OCCCOCCOCCOCCCNC(=O)O[C@@H]1CC[C@]2(C)C(=CCC3C2CC[C@]2(C)C3CCC2[C@@H](C)CCCC(C)C)C1 Chemical compound C.COCCCCCCCCCC(=O)CCCCOCC(CO)OCCCOCCOCCOCCCNC(=O)O[C@@H]1CC[C@]2(C)C(=CCC3C2CC[C@]2(C)C3CCC2[C@@H](C)CCCC(C)C)C1 VEMFGJIHJBILSW-ZVVYFNDBSA-N 0.000 description 1
- AMWRMNZJUUZXJU-BIFLFOBKSA-N CC(C)CCC[C@@H](C)[C@@H](CC1)[C@@](C)(CC2)C1C1C2[C@@](C)(CC[C@@H](C2)OC(N)=O)C2=CC1 Chemical compound CC(C)CCC[C@@H](C)[C@@H](CC1)[C@@](C)(CC2)C1C1C2[C@@](C)(CC[C@@H](C2)OC(N)=O)C2=CC1 AMWRMNZJUUZXJU-BIFLFOBKSA-N 0.000 description 1
- FFMGCGVRWLTZHH-UHFFFAOYSA-N CCC.CCC.CCC.CCC.CCC Chemical compound CCC.CCC.CCC.CCC.CCC FFMGCGVRWLTZHH-UHFFFAOYSA-N 0.000 description 1
- GMVBAMFBAFBELK-DNODHAPCSA-N CCCCCCCCCCC(=O)NCCCOCC(C)O.CCCOCCOCCOCCCNC(=O)O[C@@H]1CC[C@]2(C)C(=CCC3C2CC[C@@]2(C)C(C(C)CCCC(C)C)CCC32)C1.CCOCCOCCOCCOCCCNC(=O)O[C@H]1CC[C@@]2(C)C(=CCC3C2CC[C@@]2(C)C3CC[C@@H]2[C@H](C)CCCC(C)C)C1.COCCCCCCCCCC(=O)CCCCOCC(CO)OC Chemical compound CCCCCCCCCCC(=O)NCCCOCC(C)O.CCCOCCOCCOCCCNC(=O)O[C@@H]1CC[C@]2(C)C(=CCC3C2CC[C@@]2(C)C(C(C)CCCC(C)C)CCC32)C1.CCOCCOCCOCCOCCCNC(=O)O[C@H]1CC[C@@]2(C)C(=CCC3C2CC[C@@]2(C)C3CC[C@@H]2[C@H](C)CCCC(C)C)C1.COCCCCCCCCCC(=O)CCCCOCC(CO)OC GMVBAMFBAFBELK-DNODHAPCSA-N 0.000 description 1
- ONBDZVKBLARYKM-CZPYMLDESA-N CCCCCCCCCCC(=O)NCCCOCC(O)CCOCCOCCOCCOCCCNC(=O)O[C@H]1CC[C@@]2(C)C(=CCC3C2CC[C@@]2(C)C3CC[C@@H]2[C@H](C)CCCC(C)C)C1 Chemical compound CCCCCCCCCCC(=O)NCCCOCC(O)CCOCCOCCOCCOCCCNC(=O)O[C@H]1CC[C@@]2(C)C(=CCC3C2CC[C@@]2(C)C3CC[C@@H]2[C@H](C)CCCC(C)C)C1 ONBDZVKBLARYKM-CZPYMLDESA-N 0.000 description 1
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 description 1
- GAGWJHPBXLXJQN-UHFFFAOYSA-N Capecitabine Natural products C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1C1C(O)C(O)C(C)O1 GAGWJHPBXLXJQN-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- DLGOEMSEDOSKAD-UHFFFAOYSA-N Carmustine Chemical compound ClCCNC(=O)N(N=O)CCCl DLGOEMSEDOSKAD-UHFFFAOYSA-N 0.000 description 1
- 235000015655 Crocus sativus Nutrition 0.000 description 1
- 208000006343 Cutaneous Mastocytosis Diseases 0.000 description 1
- 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 1
- 102000011724 DNA Repair Enzymes Human genes 0.000 description 1
- 108010076525 DNA Repair Enzymes Proteins 0.000 description 1
- 108090000323 DNA Topoisomerases Proteins 0.000 description 1
- 102000003915 DNA Topoisomerases Human genes 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010012812 Diffuse cutaneous mastocytosis Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 206010014958 Eosinophilic leukaemia Diseases 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 108020004206 Gamma-glutamyltransferase Proteins 0.000 description 1
- 206010051066 Gastrointestinal stromal tumour Diseases 0.000 description 1
- 208000032320 Germ cell tumor of testis Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 208000012766 Growth delay Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 101710195517 Histone H2AX Proteins 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 1
- 101001068219 Homo sapiens Serine/threonine-protein phosphatase 4 catalytic subunit Proteins 0.000 description 1
- GRRNUXAQVGOGFE-UHFFFAOYSA-N Hygromycin-B Natural products OC1C(NC)CC(N)C(O)C1OC1C2OC3(C(C(O)C(O)C(C(N)CO)O3)O)OC2C(O)C(CO)O1 GRRNUXAQVGOGFE-UHFFFAOYSA-N 0.000 description 1
- 206010048643 Hypereosinophilic syndrome Diseases 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010061252 Intraocular melanoma Diseases 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 238000012313 Kruskal-Wallis test Methods 0.000 description 1
- 102000015335 Ku Autoantigen Human genes 0.000 description 1
- 108010025026 Ku Autoantigen Proteins 0.000 description 1
- 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 1
- 102000000853 LDL receptors Human genes 0.000 description 1
- 108010001831 LDL receptors Proteins 0.000 description 1
- 208000031671 Large B-Cell Diffuse Lymphoma Diseases 0.000 description 1
- 206010053180 Leukaemia cutis Diseases 0.000 description 1
- 206010024305 Leukaemia monocytic Diseases 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- GQYIWUVLTXOXAJ-UHFFFAOYSA-N Lomustine Chemical compound ClCCN(N=O)C(=O)NC1CCCCC1 GQYIWUVLTXOXAJ-UHFFFAOYSA-N 0.000 description 1
- 206010025557 Malignant fibrous histiocytoma of bone Diseases 0.000 description 1
- 206010051676 Metastases to peritoneum Diseases 0.000 description 1
- 108091092878 Microsatellite Proteins 0.000 description 1
- 208000035489 Monocytic Acute Leukemia Diseases 0.000 description 1
- 208000034578 Multiple myelomas Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 208000014767 Myeloproliferative disease Diseases 0.000 description 1
- 201000007224 Myeloproliferative neoplasm Diseases 0.000 description 1
- 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 description 1
- 108700019961 Neoplasm Genes Proteins 0.000 description 1
- 102000048850 Neoplasm Genes Human genes 0.000 description 1
- 208000003788 Neoplasm Micrometastasis Diseases 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 108010020346 Polyglutamic Acid Proteins 0.000 description 1
- 208000033826 Promyelocytic Acute Leukemia Diseases 0.000 description 1
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 1
- 201000000582 Retinoblastoma Diseases 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 1
- 208000033133 Testicular seminomatous germ cell tumor Diseases 0.000 description 1
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 description 1
- 102000002689 Toll-like receptor Human genes 0.000 description 1
- 108020000411 Toll-like receptor Proteins 0.000 description 1
- 102000007537 Type II DNA Topoisomerases Human genes 0.000 description 1
- 108010046308 Type II DNA Topoisomerases Proteins 0.000 description 1
- 208000015778 Undifferentiated pleomorphic sarcoma Diseases 0.000 description 1
- 206010046752 Urticaria Pigmentosa Diseases 0.000 description 1
- 201000005969 Uveal melanoma Diseases 0.000 description 1
- 102100036973 X-ray repair cross-complementing protein 5 Human genes 0.000 description 1
- 101710124921 X-ray repair cross-complementing protein 5 Proteins 0.000 description 1
- UKFDBDGJFYRBHE-FKGSWUQWSA-N [(3e,6s,7s,9s)-5-[(2r,3s)-3-acetyloxy-2-(2-hydroxyethyl)oxetan-3-yl]-3,7-dihydroxy-9-[(2r,3s)-2-hydroxy-5-methyl-3-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoyl]oxy-4,10,11,11-tetramethyl-2-oxo-6-bicyclo[5.3.1]undeca-1(10),3-dienyl] benzoate Chemical compound O([C@@H]1[C@]2(O)C[C@@H](C(=C(C(=O)\C(O)=C(C)/C1[C@]1([C@H](OC1)CCO)OC(C)=O)C2(C)C)C)OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)CC(C)C)C(=O)C1=CC=CC=C1 UKFDBDGJFYRBHE-FKGSWUQWSA-N 0.000 description 1
- SJLGWQCSCCAMBD-IBARITJRSA-N [H][C@@]12CC=C3C[C@@H](OC(=O)NCCCOCCOCCOCCOCCOP(=O)(O)OCOP(=O)(O)OCC(O)COCCCNC(=O)CCCCCCCCCCOP(=O)(O)OC)CC[C@]3(C)[C@@]1([H])CC[C@]1(C)[C@@H]([C@H](C)CCCC(C)C)CC[C@@]21[H] Chemical compound [H][C@@]12CC=C3C[C@@H](OC(=O)NCCCOCCOCCOCCOCCOP(=O)(O)OCOP(=O)(O)OCC(O)COCCCNC(=O)CCCCCCCCCCOP(=O)(O)OC)CC[C@]3(C)[C@@]1([H])CC[C@]1(C)[C@@H]([C@H](C)CCCC(C)C)CC[C@@]21[H] SJLGWQCSCCAMBD-IBARITJRSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 229940028652 abraxane Drugs 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000002487 adenosine deaminase inhibitor Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 201000006966 adult T-cell leukemia Diseases 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 229940045714 alkyl sulfonate alkylating agent Drugs 0.000 description 1
- 150000008052 alkyl sulfonates Chemical class 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 229960001220 amsacrine Drugs 0.000 description 1
- XCPGHVQEEXUHNC-UHFFFAOYSA-N amsacrine Chemical compound COC1=CC(NS(C)(=O)=O)=CC=C1NC1=C(C=CC=C2)C2=NC2=CC=CC=C12 XCPGHVQEEXUHNC-UHFFFAOYSA-N 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229940045719 antineoplastic alkylating agent nitrosoureas Drugs 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000010108 arterial embolization Effects 0.000 description 1
- 238000011717 athymic nude mouse Methods 0.000 description 1
- 238000011888 autopsy Methods 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 208000010572 basal-like breast carcinoma Diseases 0.000 description 1
- 230000033590 base-excision repair Effects 0.000 description 1
- 230000003542 behavioural effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 210000003969 blast cell Anatomy 0.000 description 1
- 201000000053 blastoma Diseases 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- UBJAHGAUPNGZFF-XOVTVWCYSA-N bms-184476 Chemical compound O([C@H]1[C@@H]2[C@]3(OC(C)=O)CO[C@@H]3C[C@@H]([C@]2(C(=O)[C@H](OC(C)=O)C2=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)C=3C=CC=CC=3)C=3C=CC=CC=3)C[C@]1(O)C2(C)C)C)OCSC)C(=O)C1=CC=CC=C1 UBJAHGAUPNGZFF-XOVTVWCYSA-N 0.000 description 1
- GMJWGJSDPOAZTP-MIDYMNAOSA-N bms-188797 Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](OC(C)=O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)C=4C=CC=CC=4)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)OC)C(=O)C1=CC=CC=C1 GMJWGJSDPOAZTP-MIDYMNAOSA-N 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 201000008274 breast adenocarcinoma Diseases 0.000 description 1
- 229960002092 busulfan Drugs 0.000 description 1
- 229960001573 cabazitaxel Drugs 0.000 description 1
- BMQGVNUXMIRLCK-OAGWZNDDSA-N cabazitaxel Chemical compound O([C@H]1[C@@H]2[C@]3(OC(C)=O)CO[C@@H]3C[C@@H]([C@]2(C(=O)[C@H](OC)C2=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=3C=CC=CC=3)C[C@]1(O)C2(C)C)C)OC)C(=O)C1=CC=CC=C1 BMQGVNUXMIRLCK-OAGWZNDDSA-N 0.000 description 1
- 229940127093 camptothecin Drugs 0.000 description 1
- 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 1
- 229960004117 capecitabine Drugs 0.000 description 1
- 229960005243 carmustine Drugs 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000010109 chemoembolization Effects 0.000 description 1
- 230000000973 chemotherapeutic effect Effects 0.000 description 1
- 231100001157 chemotherapeutic toxicity Toxicity 0.000 description 1
- 239000012829 chemotherapy agent Substances 0.000 description 1
- 238000009104 chemotherapy regimen Methods 0.000 description 1
- 229960004630 chlorambucil Drugs 0.000 description 1
- JCKYGMPEJWAADB-UHFFFAOYSA-N chlorambucil Chemical compound OC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 JCKYGMPEJWAADB-UHFFFAOYSA-N 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 208000021668 chronic eosinophilic leukemia Diseases 0.000 description 1
- 238000009643 clonogenic assay Methods 0.000 description 1
- 231100000096 clonogenic assay Toxicity 0.000 description 1
- 230000005025 clonogenic survival Effects 0.000 description 1
- 238000011278 co-treatment Methods 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 description 1
- 208000030242 cutaneous mastocytoma Diseases 0.000 description 1
- 201000006515 cutaneous solitary mastocytoma Diseases 0.000 description 1
- 125000000753 cycloalkyl group Chemical group 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 229960003901 dacarbazine Drugs 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 229960000975 daunorubicin Drugs 0.000 description 1
- 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 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 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 1
- 229960003668 docetaxel Drugs 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 201000008184 embryoma Diseases 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 210000001163 endosome Anatomy 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 230000002327 eosinophilic effect Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 229960005420 etoposide Drugs 0.000 description 1
- 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 1
- 230000003203 everyday effect Effects 0.000 description 1
- 208000021045 exocrine pancreatic carcinoma Diseases 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- ODKNJVUHOIMIIZ-RRKCRQDMSA-N floxuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ODKNJVUHOIMIIZ-RRKCRQDMSA-N 0.000 description 1
- 229960000961 floxuridine 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
- 229960005304 fludarabine phosphate Drugs 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000012632 fluorescent imaging Methods 0.000 description 1
- 239000004052 folic acid antagonist Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229960004783 fotemustine Drugs 0.000 description 1
- YAKWPXVTIGTRJH-UHFFFAOYSA-N fotemustine Chemical compound CCOP(=O)(OCC)C(C)NC(=O)N(CCCl)N=O YAKWPXVTIGTRJH-UHFFFAOYSA-N 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 102000006640 gamma-Glutamyltransferase Human genes 0.000 description 1
- 229920000370 gamma-poly(glutamate) polymer Polymers 0.000 description 1
- 229960005277 gemcitabine Drugs 0.000 description 1
- 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 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000009650 gentamicin protection assay Methods 0.000 description 1
- 201000009277 hairy cell leukemia Diseases 0.000 description 1
- 208000014951 hematologic disease Diseases 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 210000002767 hepatic artery Anatomy 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000036732 histological change Effects 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 150000007857 hydrazones Chemical class 0.000 description 1
- GRRNUXAQVGOGFE-NZSRVPFOSA-N hygromycin B Chemical compound O[C@@H]1[C@@H](NC)C[C@@H](N)[C@H](O)[C@H]1O[C@H]1[C@H]2O[C@@]3([C@@H]([C@@H](O)[C@@H](O)[C@@H](C(N)CO)O3)O)O[C@H]2[C@@H](O)[C@@H](CO)O1 GRRNUXAQVGOGFE-NZSRVPFOSA-N 0.000 description 1
- 229940097277 hygromycin b Drugs 0.000 description 1
- 230000037417 hyperactivation Effects 0.000 description 1
- 229960001101 ifosfamide Drugs 0.000 description 1
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 238000005462 in vivo assay Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000002697 interventional radiology Methods 0.000 description 1
- 229950005428 intoplicine Drugs 0.000 description 1
- QROONAIPJKQFMC-UHFFFAOYSA-N intoplicine Chemical compound C1=C(O)C=CC2=C(C=3C(NCCCN(C)C)=NC=C(C)C=3N3)C3=CC=C21 QROONAIPJKQFMC-UHFFFAOYSA-N 0.000 description 1
- 238000007917 intracranial administration Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 229960004768 irinotecan Drugs 0.000 description 1
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 description 1
- 229950005692 larotaxel Drugs 0.000 description 1
- DXOJIXGRFSHVKA-BZVZGCBYSA-N larotaxel Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@@]23[C@H]1[C@@]1(CO[C@@H]1C[C@@H]2C3)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 DXOJIXGRFSHVKA-BZVZGCBYSA-N 0.000 description 1
- 210000000867 larynx Anatomy 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000000610 leukopenic effect Effects 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 229960002247 lomustine Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 201000005249 lung adenocarcinoma Diseases 0.000 description 1
- 210000004324 lymphatic system Anatomy 0.000 description 1
- 208000025036 lymphosarcoma Diseases 0.000 description 1
- 208000030179 maculopapular cutaneous mastocytosis Diseases 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 201000006512 mast cell neoplasm Diseases 0.000 description 1
- 208000006971 mastocytoma Diseases 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229960004961 mechlorethamine Drugs 0.000 description 1
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical compound ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 210000003593 megakaryocyte Anatomy 0.000 description 1
- 229960001924 melphalan Drugs 0.000 description 1
- 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 1
- 229960001428 mercaptopurine Drugs 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- XIVMHSNIQAICTR-UQYHODNASA-N milataxel Chemical compound O([C@H]1[C@@H]2[C@]3(OC(C)=O)CO[C@@H]3C[C@@H]([C@]2(C(=O)[C@H](O)C2=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=3OC=CC=3)C[C@]1(O)C2(C)C)C)OC(=O)CC)C(=O)C1=CC=CC=C1 XIVMHSNIQAICTR-UQYHODNASA-N 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 229960001156 mitoxantrone Drugs 0.000 description 1
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 210000001167 myeloblast Anatomy 0.000 description 1
- 208000017869 myelodysplastic/myeloproliferative disease Diseases 0.000 description 1
- 230000002071 myeloproliferative effect Effects 0.000 description 1
- 230000009826 neoplastic cell growth Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000006780 non-homologous end joining Effects 0.000 description 1
- 210000004882 non-tumor cell Anatomy 0.000 description 1
- 208000026878 nongerminomatous germ cell tumor Diseases 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 201000002575 ocular melanoma Diseases 0.000 description 1
- 238000002515 oligonucleotide synthesis Methods 0.000 description 1
- 229950001094 ortataxel Drugs 0.000 description 1
- 150000002905 orthoesters Chemical class 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 229960002239 paclitaxel poliglumex Drugs 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- FPVKHBSQESCIEP-JQCXWYLXSA-N pentostatin Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC[C@H]2O)=C2N=C1 FPVKHBSQESCIEP-JQCXWYLXSA-N 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 239000008063 pharmaceutical solvent Substances 0.000 description 1
- 125000005541 phosphonamide group Chemical group 0.000 description 1
- 150000008298 phosphoramidates Chemical class 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229960000952 pipobroman Drugs 0.000 description 1
- NJBFOOCLYDNZJN-UHFFFAOYSA-N pipobroman Chemical compound BrCCC(=O)N1CCN(C(=O)CCBr)CC1 NJBFOOCLYDNZJN-UHFFFAOYSA-N 0.000 description 1
- 208000010626 plasma cell neoplasm Diseases 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 208000037821 progressive disease Diseases 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 201000001514 prostate carcinoma Diseases 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 201000006845 reticulosarcoma Diseases 0.000 description 1
- 208000029922 reticulum cell sarcoma Diseases 0.000 description 1
- 235000013974 saffron Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000033443 single strand break repair Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000011255 standard chemotherapy Methods 0.000 description 1
- 238000011301 standard therapy Methods 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 229960001052 streptozocin Drugs 0.000 description 1
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 1
- 201000010033 subleukemic leukemia Diseases 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 206010042863 synovial sarcoma Diseases 0.000 description 1
- 238000011521 systemic chemotherapy Methods 0.000 description 1
- 229960004964 temozolomide Drugs 0.000 description 1
- 229950009016 tesetaxel Drugs 0.000 description 1
- 208000002918 testicular germ cell tumor Diseases 0.000 description 1
- 208000024662 testicular seminoma Diseases 0.000 description 1
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 1
- 229960001196 thiotepa Drugs 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 229960003087 tioguanine Drugs 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002723 toxicity assay Methods 0.000 description 1
- 206010044412 transitional cell carcinoma Diseases 0.000 description 1
- 229950001353 tretamine Drugs 0.000 description 1
- IUCJMVBFZDHPDX-UHFFFAOYSA-N tretamine Chemical compound C1CN1C1=NC(N2CC2)=NC(N2CC2)=N1 IUCJMVBFZDHPDX-UHFFFAOYSA-N 0.000 description 1
- 150000004654 triazenes Chemical class 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 208000018417 undifferentiated high grade pleomorphic sarcoma of bone Diseases 0.000 description 1
- 238000011870 unpaired t-test Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- AHXICHPPXIGCBN-GPWPDEGDSA-N uqc681jjiv Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](OC(C)=O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C(C)(C)C)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)OC)C(=O)C1=CC=CC=C1 AHXICHPPXIGCBN-GPWPDEGDSA-N 0.000 description 1
- 229960001055 uracil mustard Drugs 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 208000023747 urothelial carcinoma Diseases 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 229960002066 vinorelbine Drugs 0.000 description 1
- GBABOYUKABKIAF-GHYRFKGUSA-N vinorelbine Chemical compound C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC GBABOYUKABKIAF-GHYRFKGUSA-N 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 208000016261 weight loss Diseases 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 238000002689 xenotransplantation Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/555—Heterocyclic compounds containing heavy metals, e.g. hemin, hematin, melarsoprol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/243—Platinum; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- 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
- A61P35/04—Antineoplastic agents specific for metastasis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/13—Decoys
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/31—Chemical structure of the backbone
- C12N2310/315—Phosphorothioates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/31—Chemical structure of the backbone
- C12N2310/318—Chemical structure of the backbone where the PO2 is completely replaced, e.g. MMI or formacetal
- C12N2310/3183—Diol linkers, e.g. glycols or propanediols
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
- C12N2310/3515—Lipophilic moiety, e.g. cholesterol
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/50—Physical structure
- C12N2310/53—Physical structure partially self-complementary or closed
- C12N2310/531—Stem-loop; Hairpin
Definitions
- the present invention relates to the field of oncology.
- Breast cancer is the most common female malignancy, with more than 1.7 million new cases diagnosed each year worldwide (Torre, Siegel, Ward, & Jemal, 2015). Molecular classification divides breast cancer in three major sub-groups: LuminalA/B, HER2 + and Basal-like/Triple negative breast cancer (TNBC)(Vuong, Simpson, Green, Cummings, & Lakhani, 2014). Triple-negative breast cancer (TN BC) is an aggressive histological subtype with limited treatment options and very poor prognosis following progression after standard chemotherapeutic regimens.
- Chemo-resistance presents a major obstacle to the efficacy of cancer treatment.
- DNA repair plays a key role in chemo-resistance by eliminating the damage induced on chromosomes by the chemotherapeutic agents and inhibitors of DNA repair pathways may provide novel opportunities for restoring tumor sensitivity to these treatments.
- Dbait molecules are a new class of DNA repair inhibitors triggering false DNA damage signaling in cancer cells. These molecules are short double-stranded DNA with a free double strand blunt end, which target key damage signal transducers such as DNA dependent protein kinase (DNA-PK) and Poly-ADP-Ribo-Polymerase, triggering their activation and amplifying false damage signaling. Consequently, the recruitment of downstream DNA repair enzymes is impaired, inhibiting several DNA repair pathways such as homologous recombination, non-homologous end joining, base excision repair and single-strand break repair leading to an accumulation of unrepaired damage causing cell death.
- DNA-PK DNA dependent protein kinase
- Poly-ADP-Ribo-Polymerase Poly-ADP-Ribo-Polymerase
- Dbait molecules have been shown to be effective in combination with radiotherapy on several radio-resistant tumors, both in vitro and in vivo.
- the Dbait molecule was modified by covalently linking a cholesterol moiety to the 5′-end (DT01) (WO2011/161075; Berthault et al, 2011, Cancer gene therapy, 18, 695-706). It has been demonstrated that local administration of DT01 by intra-tumoral injection in association with radiotherapy, increases survival of xenografted human melanoma models (Biau et al, 2014, Neoplasia, 16, 835-844). However to date, the efficacy of systemic administration of DT01, alone or in association with chemotherapy, has not been investigated.
- TACE transarterial chemoembolization
- the molecule DT01 was designed for being used in combination with an endosomolytic agent such as the chloroquine.
- the chloroquine facilitates the release of coDbait from endosomes into the cytosol and is described as necessary. It was thus shown that coDbait administered by subcutaneous and intratumoral injection in the presence of chloroquine and that concomitant chloroquine treatment was added to the protocol to increase coDbait uptake and efficacy resulting in an increase of the radiosensitivity of xenografted tumors by coDbait (Schlegel et al, 2012, Molecular Therapy—Nucleic Acids, 1, e33).
- TNBC triple-negative breast cancer
- the inventors observed that the DBait molecules, especially those called coDBait which are conjugated to cholesterol, can be used efficiently for treating cancer, even resistant cancer, by systemic administration, especially intraperitoneal and intravenous administration, without any quinoline endosomlytic agent, especially chloroquine. Indeed, the same efficacy by these administration routes can be obtained with a dose which is only 2-5 fold higher and conjugated DBait has no toxicity. The efficacy of the conjugated DBait with intraperitoneal and intravenous administration has been shown with resistant cancers.
- the present invention relates to a nucleic acid molecule for use for treating cancer, wherein the nucleic acid molecule has one of the following formula:
- N is a deoxynucleotide
- n is an integer from 15 to 195
- the underlined N refers to a nucleotide having or not a modified phosphodiester backbone
- L′ is a linker
- C is the molecule facilitating endocytosis selected from a lipophilic molecule or a ligand which targets cell receptor enabling receptor mediated endocytosis
- L is a linker
- m is an integer being 0 or 1 and p is 1;
- nucleic acid is to be used without combined administration of an endosomolytic agent
- nucleic acid is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes.
- nucleic acid of formula (I) has one or several of the following features:
- nucleic acid molecule has one of the following formulae:
- underlined nucleotide refers to a nucleotide having or not a phosphorothioate or methylphosphonate backbone
- the linked L′ is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane
- m is 1
- L is a carboxamido oligoethylene glycol
- C is selected from the group consisting of dioleoyl, octadecyl, folic acid, tocopherol and cholesterol;
- nucleic acid is to be used without any combined administration of an endosomolytic agent, especially chloroquine;
- nucleic acid is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes.
- nucleic acid 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-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-aminoethyl)-2-aminoethyl-N
- underlined nucleotide refers to a nucleotide having a phosphorothioate backbone.
- the nucleic acid is to be administered by intravenous route, for instance administered by injection, intravenous drip, bolus or pump.
- the cancer is a radioresistant or chemoresistant cancer. More preferably, it is selected from the group consisting of a triple-negative breast cancer (TNBC), a chemoresistant hepatocellular carcinoma (HCC), a chemoresistant ovarian cancer, a chemoresistant lung cancer, and a metastatic liver cancer.
- TNBC triple-negative breast cancer
- HCC chemoresistant hepatocellular carcinoma
- chemoresistant ovarian cancer a chemoresistant lung cancer
- a metastatic liver cancer a particular embodiment, the cancer is selected from the group consisting of a doxorubicin-resistant hepatocarcinoma (HCC), a platinum-resistant ovarian cancer, a platinum-resistant triple-negative breast cancer and a colorectal liver metastasis.
- the nucleic acid is to be used in combination with radiotherapy and/or chemotherapy.
- the nucleic acid is to be used in combination with a DNA damaging agent.
- the DNA damaging agent is selected from the group consisting of an inhibitor of topoisomerases I or II, a DNA crosslinker, a DNA alkylating agent, an anti-metabolic agent and inhibitors of the mitotic spindles.
- the nucleic acid is to be used in combination with a chemotherapy selected from the group consisting of doxorubicin, oxaliplatin, carboplatin, cisplatin and 5-FU.
- the present invention relates to a nucleic acid molecule (coDBait) for use for treating cancer, wherein the nucleic acid molecule has one of the following formula:
- N is a deoxynucleotide
- n is an integer from 1 to 195
- the underlined N refers to a nucleotide having or not a modified phosphodiester backbone
- L′ is a linker
- C is the molecule facilitating endocytosis selected from a lipophilic molecule or a ligand which targets cell receptor enabling receptor mediated endocytosis
- L is a linker
- m is an integer being 0 or 1 and p is 1;
- nucleic acid is to be used without combined administration of an endosomolytic agent
- nucleic acid is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes.
- quinoline endosomolytic agent refers to the endosomolytic agent as defined WO2011/161075 in pages 26-28 of (incorporated herein by reference).
- the quinoline endosomolytic agent is the chloroquine.
- nucleic acid as described herein is not for simultaneous, separate or sequential use with any endosomolytic agent.
- the present invention relates to
- kit defines especially a “kit of parts” in the sense that the combination partners as defined above can be dosed independently or by use of different fixed combinations with distinguished amounts of the combination partners, i.e. simultaneously or at different time points.
- the parts of the kit of parts can then, e.g., be administered simultaneously or chronologically staggered, that is at different time points and with equal or different time intervals for any part of the kit of parts.
- the ratio of the total amounts of the combination partners to be administered in the combined preparation can be varied.
- the combination partners can be administered by the same route or by different routes.
- the term treatment denotes curative, symptomatic, and preventive treatment.
- Pharmaceutical compositions, kits, products and combined preparations of the invention can be used in humans with existing cancer or tumor, including at early or late stages of progression of the cancer.
- the pharmaceutical compositions, kits, products and combined preparations of the invention will not necessarily cure the patient who has the cancer but will delay or slow the progression or prevent further progression of the disease, ameliorating thereby the patients' condition.
- the pharmaceutical compositions, kits, products and combined preparations of the invention reduce the development of tumors, reduce tumor burden, produce tumor regression in a mammalian host and/or prevent metastasis occurrence and cancer relapse.
- the pharmaceutical composition of the invention is administered in a therapeutically effective amount.
- therapeutically effective amount it is meant the quantity of the pharmaceutical composition of the invention which prevents, removes or reduces the deleterious effects of cancer in mammals, including humans, alone or in combination with the other active ingredients of the pharmaceutical composition, kit, product or combined preparation. It is understood that the administered dose may be lower for each compound in the composition to the “therapeutic effective amount” define for each compound used alone or in combination with other treatments than the combination described here.
- the “therapeutic effective amount” of the composition will be adapted by those skilled in the art according to the patient, the pathology, the mode of administration, etc.
- treatment of a cancer or the like is mentioned with reference to the pharmaceutical composition of the invention, there is meant: a) a method for treating a cancer, said method comprising administering a pharmaceutical composition of the invention to a subject in need of such treatment; b) the use of a pharmaceutical composition of the invention for the treatment of a cancer; c) the use of a pharmaceutical composition of the invention for the manufacture of a medicament for the treatment of a cancer; and/or d) a pharmaceutical composition of the invention for use in the treatment a cancer.
- compositions contemplated herein may include a pharmaceutically acceptable carrier in addition to the active ingredient(s).
- pharmaceutically acceptable carrier is meant to encompass any carrier (e.g., support, substance, solvent, etc.) which does not interfere with effectiveness of the biological activity of the active ingredient(s) and that is not toxic to the host to which it is administered.
- the active compounds(s) may be formulated in a unit dosage form for injection in vehicles such as saline, dextrose solution, serum albumin and Ringer's solution.
- the pharmaceutical composition can be formulated as solutions in pharmaceutically compatible solvents or as emulsions, suspensions or dispersions in suitable pharmaceutical solvents or vehicle in a way known in the art.
- Formulations suitable for parental administration conveniently comprise a sterile oily or aqueous preparation of the active ingredient which is preferably isotonic with the blood of the recipient. Every such formulation can also contain other pharmaceutically compatible and nontoxic auxiliary agents, such as, e.g. stabilizers, antioxidants, binders, dyes, emulsifiers or flavouring substances.
- the formulations of the present invention comprise an active ingredient in association with a pharmaceutically acceptable carrier therefore and optionally other therapeutic ingredients.
- the carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient thereof.
- the pharmaceutical compositions are advantageously applied by injection or intravenous infusion of suitable sterile solutions. Methods for the safe and effective administration of most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in the standard literature.
- DBait molecules for use in the present invention can be described by the following formulae:
- N is a nucleotide
- n is an integer of at least 1
- the underlined N refers to a nucleotide having or not a modified phosphodiester backbone
- L′ is a linker
- C is a molecule facilitating endocytosis
- L is a linker
- m is an integer being 0 or 1
- p is 1.
- the underlined N refers to a nucleotide having a modified phosphodiester backbone.
- the molecule of formula (I) has one or several of the following features:
- C-Lm is a triethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-triethyleneglycol radical).
- C-Lm is a tetraethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-tetraethyleneglycol radical).
- the nucleic acid molecules can be Dbait molecules such as those extensively described in PCT patent applications WO2005/040378, WO2008/034866 and WO2008/084087, the disclosure of which is incorporated herein by reference.
- Dbait molecules may be defined by a number of characteristics necessary for their therapeutic activity, such as their minimal length, the presence of at least one free end, and the presence of a double stranded portion, preferably a DNA double stranded portion. As will be discussed below, it is important to note that the precise nucleotide sequence of Dbait molecules does not impact on their activity. Furthermore, Dbait molecules may contain a modified and/or non-natural backbone.
- Dbait molecules are of non-human origin (i.e., their nucleotide sequence and/or conformation (e.g., hairpin) does not exist as such in a human cell), most preferably of synthetic origin.
- sequence of the Dbait molecules plays little, if any, role, Dbait molecules have preferably no significant degree of sequence homology or identity to known genes, promoters, enhancers, 5′- or 3′-upstream sequences, exons, introns, and the like.
- Dbait molecules have less than 80% or 70%, even less than 60% or 50% sequence identity to any gene in a human genome. Methods of determining sequence identity are well known in the art and include, e.g., Blast.
- Dbait molecules do not hybridize, under stringent conditions, with human genomic DNA. Typical stringent conditions are such that they allow the discrimination of fully complementary nucleic acids from partially complementary nucleic acids.
- sequence of the Dbait molecules is preferably devoid of CpG in order to avoid the well-known toll-like receptor-mediated immunological reactions.
- the length of Dbait molecules may be variable, as long as it is sufficient to allow appropriate binding of Ku protein complex comprising Ku and DNA-PKcs proteins. It has been showed that the length of Dbait molecules must be greater than 20 bp, preferably about 32 bp, to ensure binding to such a Ku complex and allowing DNA-PKcs activation.
- Dbait molecules comprise between 20-200 bp, more preferably 24-100 bp, still more preferably 26-100, and most preferably between 24-200, 25-200, 26-200, 27-200, 28-200, 30-200, 32-200, 24-100, 25-100, 26-100, 27-100, 28-100, 30-100, 32-200 or 32-100 bp.
- Dbait molecules comprise between 24-160, 26-150, 28-140, 28-200, 30-120, 32-200 or 32-100 bp.
- bp is intended that the molecule comprise a double stranded portion of the indicated length.
- the Dbait molecules having a double stranded portion of at least 32 pb, or of about 32 bp comprise the same nucleotide sequence than Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- the Dbait molecules have the same nucleotide composition than Dbait32, Dbait32Ha, Dbait32Hb, Dbait32Hc or Dbait32Hd but their nucleotide sequence is different.
- the Dbait molecules comprise one strand of the double stranded portion with 3 A, 6 C, 12 G and 11 T.
- the sequence of the Dbait molecules does not contain any CpG dinucleotide.
- the double stranded portion comprises at least 16, 18, 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- the double stranded portion consists in 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- the nucleic acid as disclosed herein must have at least one free end, as a mimic of DSB.
- Dbait molecules are made of hairpin nucleic acids with a double-stranded DNA stem and a loop.
- the loop can be a nucleic acid, or other chemical groups known by skilled person or a mixture thereof.
- a nucleotide linker may include from 2 to 10 nucleotides, preferably, 3, 4 or 5 nucleotides.
- Non-nucleotide linkers non exhaustively include abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, or other polymeric compounds (e. g.
- oligoethylene glycols such as those having between 2 and 10 ethylene glycol units, preferably 4, 5, 6, 7 or 8 ethylene glycol units).
- a preferred linker is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and other linkers such as 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.
- the Dbait molecules can be a hairpin molecule having a double stranded portion or stem comprising at least 16, 18, 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5) and a loop being a hexaethyleneglycol linker, a tetradeoxythymidylate linker (T4) or 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.
- Dbait32 SEQ ID No 1
- Dbait32Ha SEQ ID No 2
- Dbait32Hb SEQ ID No 3
- Dbait32Hc SEQ ID No 4
- Dbait32Hd SEQ ID No 5
- T4 tetradeoxythymidylate
- those Dbait molecules can have a double stranded portion consisting in 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- Dbait molecules preferably comprise a 2′-deoxynucleotide backbone, and optionally comprise one or several (2, 3, 4, 5 or 6) modified nucleotides and/or nucleobases other than adenine, cytosine, guanine and thymine. Accordingly, the Dbait molecules are essentially a DNA structure. In particular, the double-strand portion or stem of the Dbait molecules is made of deoxyribonucleotides.
- Preferred Dbait molecules comprise one or several chemically modified nucleotide(s) or group(s) at the end of one or of each strand, in particular in order to protect them from degradation.
- the free end(s) of the Dbait molecules is(are) protected by one, two or three modified phosphodiester backbones at the end of one or of each strand.
- Preferred chemical groups, in particular the modified phosphodiester backbone comprise phosphorothioates.
- preferred Dbait have 3′- 3′ nucleotide linkage, or nucleotides with methylphosphonate backbone.
- modified backbones are well known in the art and comprise phosphoramidates, morpholino nucleic acid, 2′-0,4′-C methylene/ethylene bridged locked nucleic acid, peptide nucleic acid (PNA), and short chain alkyl, or cycloalkyl intersugar linkages or short chain heteroatomic or heterocyclic intrasugar linkages of variable length, or any modified nucleotides known by skilled person.
- the Dbait molecules have the free end(s) protected by one, two or three modified phosphodiester backbones at the end of one or of each strand, more preferably by three modified phosphodiester backbones (in particular phosphorothioate or methylphosphonate) at least at the 3′end, but still more preferably at both 5′ and 3′ ends.
- the Dbait molecule is a hairpin nucleic acid molecule comprising a DNA double-stranded portion or stem of 32 bp (e.g., with a sequence selected from the group consisting of SEQ ID Nos 1-5, in particular SEQ ID No 4) and a loop linking the two strands of the DNA double-stranded portion or stem comprising or consisting of a linker selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane, the free ends of the DNA double-stranded portion or stem (i.e. at the opposite of the loop) having three modified phosphodiester backbones (in particular phosphorothioate internucleotidic links).
- a linker selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19
- Said nucleic acid molecules are made by chemical synthesis, semi-biosynthesis or biosynthesis, any method of amplification, followed by any extraction and preparation methods and any chemical modification.
- Linkers are provided so as to be incorporable by standard nucleic acid chemical synthesis.
- nucleic acid molecules are manufactured by specially designed convergent synthesis: two complementary strands are prepared by standard nucleic acid chemical synthesis with the incorporation of appropriate linker precursor, after their purification, they are covalently coupled together.
- the molecules facilitating endocytosis are conjugated to Dbait molecules, preferably through a linker.
- Any linker known in the art may be used to covalently attach the molecule facilitating endocytosis to Dbait molecules
- WO09/126933 provides a broad review of convenient linkers pages 38-45.
- the linker can be non-exhaustively, aliphatic chain, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, or other polymeric compounds (e. g.
- oligoethylene glycols such as those having between 2 and 10 ethylene glycol units, preferably 3, 4, 5, 6, 7 or 8 ethylene glycol units, still more preferably 6 ethylene glycol units), as well as incorporating any bonds that may be break down by chemical or enzymatical way, such as a disulfide linkage, a protected disulfide linkage, an acid labile linkage (e.g., hydrazone linkage), an ester linkage, an ortho ester linkage, a phosphonamide linkage, a biocleavable peptide linkage, an azo linkage or an aldehyde linkage.
- cleavable linkers are detailed in WO2007/040469 pages 12-14, in WO2008/022309 pages 22-28.
- the nucleic acid molecule can be linked to one molecule facilitating endocytosis.
- several molecules facilitating endocytosis e.g., two, three or four
- the linker between the molecule facilitating endocytosis, in particular cholesterol, and nucleic acid molecule is CO—NH—(CH 2 —CH 2 —O) n , wherein n is an integer from 1 to 10, preferably n being selected from the group consisting of 3, 4, 5 and 6.
- the linker is CO—NH—(CH 2 —CH 2 —O) 4 (carboxamido tetraethylene glycol) or CO—NH—(CH 2 —CH 2 —O) 3 (carboxamido triethylene glycol).
- the linker can be linked to nucleic acid molecules at any convenient position which does not modify the activity of the nucleic acid molecules.
- the linker can be linked at the 5′ end. Therefore, in a preferred embodiment, the contemplated conjugated Dbait molecule is a Dbait molecule having a hairpin structure and being conjugated to the molecule facilitating endocytosis, preferably through a linker, at its 5′ end.
- the linker between the molecule facilitating endocytosis, in particular cholesterol, and nucleic acid molecule is dialkyl-disulfide ⁇ e.g., (CH 2 ) r —S—S—(CH 2 ) s with r and s being integer from 1 to 10, preferably from 3 to 8, for instance 6 ⁇ .
- the conjugated Dbait molecule is a hairpin nucleic acid molecule comprising a DNA double-stranded portion or stem of 32 bp (e.g., with a sequence selected from the group consisting of SEQ ID Nos 1-5, in particular SEQ ID No 4) and a loop linking the two strands of the DNA double-stranded portion or stem comprising or consisting of a linker selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane, the free ends of the DNA double-stranded portion or stem (i.e.
- a linker e.g. carboxamido oligoethylene glycol, preferably carboxamido triethylene or tetraethylene glycol.
- NNNN—(N) n —N comprises at least 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb
- Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5) or consists in 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- NNNN—(N) n —N comprises or consists in Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5), more preferably Dbait32Hc (SEQ ID No 4).
- conjugated Dbait molecule or hairpin nucleic acid molecule may be selected from the group consisting of:
- the molecule of formulae (Ia), (Ib), (Ic), (Id), and (Ie) has one or several of the following features:
- C-Lm is a triethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-triethyleneglycol radical) or a tetraethyleneglycol linker (10-)-[1-propyl-3-N-carbamoylcholesteryl]-tetraethyleneglycol radical).
- L′ is preferably selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.
- C-L m is the radical
- the conjugated Dbait molecule or hairpin nucleic acid molecule is selected from the group consisting of (I), (Ia), (Ib), (Ic), (Id), and (Ie), wherein C-L m being the radical
- L′ is preferably selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane, more preferably 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.
- the Dbait molecule or hairpin nucleic acid molecule has the following formula
- C-L m is a tetraethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-tetraethyleneglycol radical, and L′ is 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane and wherein the underlined nucleotides have a phosphorothioate backbone.
- the molecule has the following structure and it is referred thereto in the Example section as “coDbait” or “DT01”.
- the nucleic acid molecule has one of the following formulae
- N is a deoxynucleotide
- n is an integer from 1 to 195
- the underlined N refers to a nucleotide having or not a modified phosphodiester backbone
- L′ is a linker
- C is a cholesterol
- L is a linker
- m is an integer being 0 or 1
- p is 1.
- the underlined N refers to a nucleotide having a modified phosphodiester backbone.
- the treatment may also further comprise an antitumor treatment, preferably a treatment by a DNA damaging agent or radiotherapy.
- the DNA-damaging treatment can be radiotherapy or chemotherapy with a DNA-damaging antitumor agent, or a combination thereof.
- Radiotherapy includes, but is not limited to, ⁇ -rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells.
- Other radiotherapies include microwaves and UV-irradiation.
- Other approaches to radiation therapy are also contemplated in the present invention.
- the DNA-damaging antitumor agent is preferably selected from the group consisting of an inhibitor of topoisomerases I or II, a DNA crosslinker, a DNA alkylating agent, an anti-metabolic agent and inhibitors of the mitotic spindles.
- Inhibitors of topoisomerases I and/or II include, but are not limited to, etoposide, topotecan, camptothecin, irinotecan, amsacrine, intoplicine, anthracyclines such as doxorubicin, epirubicine, daunorubicine, idanrubicine and mitoxantrone.
- Inhibitors of Topoisomerase I and II include, but are not limited to, intoplecin.
- the DNA-damaging antitumor agent is doxorubicin.
- DNA crosslinkers include, but are not limited to, cisplatin, carboplatin and oxaliplatin.
- the DNA-damaging antitumor agent is selected from the group consisting of carboplatin and oxaliplatin.
- Anti-metabolic agents block the enzymes responsible for nucleic acid synthesis or become incorporated into DNA, which produces an incorrect genetic code and leads to apoptosis.
- Non-exhaustive examples thereof include, without limitation, folic acid antagonists, pyrimidine analogs, purine analogs and adenosine deaminase inhibitors, and more particularly Methotrexate, Floxuridine, Cytarabine, 6-Mercaptopurine, 6- Thioguanine, Fludarabine phosphate, Pentostatine, 5-fluorouracil (5-FU), gemcitabine and capecitabine.
- the DNA-damaging anti-tumor agent can be alkylating agents including, without limitation, nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, metal salts and triazenes.
- Non-exhaustive examples thereof include Uracil mustard, Chlormethine, Cyclophosphamide (CYTOXAN(R)), Ifosfamide, Melphalan, Chlorambucil, Pipobroman, Triethylenemelamine, Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine, Fotemustine, cisplatin, carboplatin, oxaliplatin, thiotepa, Streptozocin, dacarbazine, and Temozolomide.
- Inhibitors of the mitotic spindles include, but are not limited to, paclitaxel, docetaxel, vinorelbine, larotaxel (also called XRP9881; Sanofi-Aventis), XRP6258 (Sanofi-Aventis), BMS-184476 (Bristol-Meyer-Squibb), BMS-188797 (Bristol-Meyer-Squibb), BMS-275183 (Bristol-Meyer-Squibb), ortataxel (also called IDN 5109, BAY 59-8862 or SB-T-101131 ; Bristol-Meyer-Squibb), RPR 109881A (Bristol-Meyer-Squibb), RPR 116258 (Bristol-Meyer-Squibb), NBT-287 (TAPESTRY), PG-paclitaxel (also called CT-2103, PPX, paclitaxel poliglumex, paclitaxel polygluta
- the DNA-damaging antitumor agent is an inhibitor of topoisomerases I and/or II, a DNA crosslinker, an anti-metabolic agent or a combination thereof.
- the DNA-damaging antitumor agent is selected from the group consisting of doxorubicin, 5-FU, carboplatin and oxaliplatin or a combination thereof.
- the conjugated DBait is DT01 and the DNA-damaging antitumor agent is selected from the group consisting of doxorubicin, carboplatin, 5-FU and oxaliplatin.
- compositions and the products, kits or combined preparation described in the invention can be used for treating cancer in a subject.
- cancer refers to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth.
- examples of cancer include, for example, leukemia, lymphoma, blastoma, carcinoma and sarcoma.
- cancers include chronic myeloid leukemia, acute lymphoblastic leukemia, Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ALL), squamous cell carcinoma, small-cell lung cancer, non-small cell lung cancer, glioma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon carcinoma, and head and neck cancer, gastric cancer, germ cell tumor, pediatric sarcoma, sinonasal natural killer, multiple myeloma, acute myelogenous leukemia (AML), chronic lymphocytic leukemia, mastocytosis and any symptom associated with mastocytosis.
- AML acute myelogenous leukemia
- Leukemia refers to progressive, malignant diseases of the blood-forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease—acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number of abnormal cells in the blood—leukemic or aleukemic (subleukemic).
- Leukemia includes, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell le
- carcinoma including that of the bladder (including accelerated and metastatic bladder cancer), breast, colon (including colorectal cancer), kidney, liver, lung (including small and non-small cell lung cancer and lung adenocarcinoma), ovary, prostate, testes, genitourinary tract, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), esophagus, stomach, gall bladder, cervix, thyroid, and skin (including squamous cell carcinoma); hematopoietic tumors of lymphoid lineage including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, histiocytic lymphoma, and Burkett
- disorders include urticaria pigmentosa, mastocytosises such as diffuse cutaneous mastocytosis, solitary mastocytoma in human, as well as dog mastocytoma and some rare subtypes like bullous, erythrodermic and teleangiectatic mastocytosis, mastocytosis with an associated hematological disorder, such as a myeloproliferative or myelodysplastic syndrome, or acute leukemia, myeloproliferative disorder associated with mastocytosis, mast cell leukemia, in addition to other cancers.
- mastocytosises such as diffuse cutaneous mastocytosis, solitary mastocytoma in human, as well as dog mastocytoma and some rare subtypes like bullous, erythrodermic and teleangiectatic mastocytosis
- mastocytosis with an associated hematological disorder such as a myeloproliferative or myelodysplastic syndrome, or acute leukemia,
- carcinoma including that of the bladder, urothelial carcinoma, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid, testis, particularly testicular seminomas, and skin; including squamous cell carcinoma; gastrointestinal stromal tumors (“GIST”); hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma and Burketts lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma and rhab
- the cancer is a solid tumor.
- solid tumor especially means breast cancer, ovarian cancer, cancer of the colon and generally the GI (gastro-intestinal) tract, cervix cancer, lung cancer, in particular small- cell lung cancer, and non-small-cell lung cancer, head and neck cancer, bladder cancer, cancer of the prostate or Kaposi's sarcoma.
- compositions and the products, kits or combined preparations described in the invention may be useful for inhibiting the growth of solid tumors, decreasing the tumor volume, preventing the metastatic spread of tumors and the growth or development of micrometastases.
- the pharmaceutical compositions and the products, kits or combined preparations described in the invention are in particular suitable for the treatment of poor prognosis patients or of radio- or chemo-resistant tumors.
- the cancer can be selected from melanoma, glioblastoma, breast cancer, colon cancer, lung cancer, gastrointestinal cancer, liver cancer and head and neck cancer.
- the cancer is a radioresistant or chemoresistant cancer. More particularly, the cancer is selected from the group consisting of a radioresistant melanoma, a triple-negative breast cancer (TNBC), a chemoresistant hepatocellular carcinoma (HCC), a chemoresistant lung cancer, a chemoresistant ovarian cancer and a metastatic liver cancer. More specifically, the cancer is selected from the group consisting of a doxorubicin-resistant hepatocarcinoma (HCC), a platinum-resistant triple-negative breast cancer, a platinum-resistant ovarian cancer and a colorectal liver metastasis.
- TNBC triple-negative breast cancer
- HCC chemoresistant hepatocellular carcinoma
- lung cancer a chemoresistant lung cancer
- a chemoresistant ovarian cancer a metastatic liver cancer.
- the cancer is selected from the group consisting of a doxorubicin-resistant hepatocarcinoma (HCC), a platinum-resistant triple-
- the present invention relates to the use of conjugated DBait molecules for the treatment of colorectal carcinoma in combination with oxaliplatin and 5-FU.
- the colorectal carcinoma is metastatic, more preferably with metastasis in the liver and/or peritoneum.
- the present invention relates to the use of conjugated DBait molecules for the treatment of radioresistant or chemoresistant cancer which is not localized into the abdominal cavity.
- Such cancers do not encompass colorectal carcinoma (e.g. CRC metastasis of the liver and the peritoneum) accordingly.
- the cancer is selected from the group consisting of a TNBC and a chemoresistant ovarian cancer. More specifically, the cancer is selected from the group consisting of a platinum-resistant TNBC and a platinum-resistant ovarian cancer.
- the present invention relates to the use of conjugated DBait molecules for the treatment of triple negative breast cancer. More specifically, the conjugated DBait molecules is used in combination with platinum-containing anti-cancer drugs, especially selected from cisplatin, oxaliplatin and carboplatin. In a specific embodiment, the present invention relates to the use of conjugated DBait molecules for the treatment of triple-negative breast cancer (TN BC) in combination with carboplatin. Optionally, the treatment can be combined with radiotherapy.
- TN BC triple-negative breast cancer
- the treatment can be combined with radiotherapy.
- each of the combination partners employed in the combined preparation of the invention may vary depending on the particular compound or pharmaceutical composition employed, the mode of administration, the condition being treated, the severity of the condition being treated.
- the dosage regimen of the combined preparation of the invention is selected in accordance with a variety of factors including the route of administration and the patient status.
- a physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the single active ingredients required to prevent, counter or arrest the progress of the condition.
- Optimal precision in achieving concentration of the active ingredients within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the active ingredients' availability to target sites.
- the DNA-damaging antitumor agent and the conjugated Dbait molecules may be administered by the same route or by distinct routes.
- the administration route for the DNA-damaging antitumor agent may be oral, parenteral, intravenous, intratumoral, subcutaneous, intracranial, intraartery, topical, rectal, transdermal, intradermal, nasal, intramuscular, intraosseous, and the like.
- the conjugated Dbait molecules is to be administered before and/or simultaneously with and/or after the irradiation and/or the administration of the DNA-damaging antitumor agent, more preferably before and/or simultaneously with the irradiation and/or the administration of the DNA-damaging antitumor agent.
- the irradiation and/or the administration of the DNA-damaging antitumor agent is performed so as the conjugated Dbait molecules are present in the tumoral cells when the irradiation is applied or when the DNA-damaging antitumor agent reach the tumoral cells.
- the physician, clinician or veterinarian of ordinary skill can determine the regimen based on the active ingredients, their kinetics of availability to target sites or their pharmacokinetic profiles in plasma.
- the irradiation is to be applied or the DNA-damaging antitumor agent is to be administered at the beginning of the treatment with conjugated Dbait molecules or after the treatment with conjugated Dbait molecules.
- the irradiation is to be applied or the DNA-damaging antitumor agent is to be administered 3-24 h after the beginning of the treatment with conjugated Dbait molecules.
- the DNA-damaging antitumor agent and conjugated Dbait molecules may also be simultaneously administered.
- the treatment with the conjugated Dbait molecules can continue as long as the treatment by radiotherapy or with the DNA-damaging antitumor agent is to be applied or administered.
- the treatment with the conjugated Dbait molecules can also end.
- the effective dosage of the DNA-damaging antitumor agent employed in the combined preparation, kit or product of the invention may vary depending on the mode of administration, the condition being treated, the severity of the condition being treated.
- the dosage regimen of the conjugated Dbait molecules is selected in accordance with a variety of factors including the route of administration and the patient status.
- a physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the conjugated Dbait molecules required to prevent, counter or arrest the progress of the cancer, in particular in combination with the selected DNA damaging treatment.
- the one skilled in the art can adapt the amount in order to obtain an efficient amount of the conjugated Dbait molecules in the tumor of at least 0.01 mg per 1 cm 3 of tumor, preferably 0.1 - 40 mg per 1 cm 3 of tumor, most preferably 1 - 20 mg per 1 cm 3 of tumor, in particular in a daily treatment protocol or in a weekly treatment protocol.
- the efficient amount or unit dosage of the conjugated Dbait molecules may be of 0.1 to 100 mg, preferably of 4 to 40 mg.
- the efficient amount or unit dosage of the conjugated Dbait molecules may be of 0.06 to 0.6 mg/kg of patient.
- the dosage and the regimen can be adapted by the one skilled in art in consideration of the chemotherapy and/or radiotherapy regimen.
- any radiotherapy regimen known in the art may be used, in particular stereotactic irradiation (e.g., 15 Gy) or a fractionated irradiation.
- the use of a fractionated irradiation may be particularly efficient, for instance irradiation may applied every day or every 2-5 days, preferably every 3-4 days, in a period of one, two, three, four, five or six weeks.
- the irradiation may be from 1 to 10 Gy, preferably from 2 to 5 Gy, in particular 2, 3, 4 or 5 Gy.
- fractionated irradiation of 15 ⁇ 2Gy in six weeks, or of 4 to 6 ⁇ 5Gy in two weeks may be contemplated.
- the contemplated radiotherapy is a protocol with 4 irradiations of 5 Gy in two weeks.
- Different regimens or conditions of combined treatments of cancer with irradiation and Dbait molecules have been tested and allowed to demonstrate the radio-sensibilization of tumors by Dbait molecules depends on the doses of Dbait molecules but not of the irradiation doses.
- the effective dosage of the DNA-damaging antitumor agent employed in the combined preparation, kit or product of the invention or in combination with the composition of the invention may vary depending on the particular DNA-damaging antitumor agent employed, the mode of administration, the condition being treated, the severity of the condition being treated.
- the dosage regimen of the DNA-damaging antitumor agent is selected in accordance with a variety of factors including the route of administration and the patient status.
- a physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the DNA-damaging antitumor agent required to prevent, counter or arrest the progress of the cancer.
- the treatment may include one or several cycles, for instance two to ten cycles, in particular two, three, four or five cycles.
- the cycles may be continued or separated. For instance, each cycle is separated by a period of time of one to eight weeks, preferably three to four weeks.
- FIG. 1 DT01 significantly increases sensitivity to CT in a CRC (HT29) liver metastatic model
- Intrahepatic tumor bearing NMRI NU/NU mice were treated as described in the Material and Methods, and sacrificed 22 days post treatment. Livers were sampled for macroscopic and microscopic examination.
- A Sequence of DT01 and CT therapy. CT was administered 2 or 4 hours post DT01 treatment.
- B Mean liver tumor volume (mm 3 ) in each treatment group.
- C Representative macroscopic and HES sections of liver tumors in each group.
- D Tumor necrotic component assessed through HES staining. Necrosis is expressed as a proportion (%) of the total tumor surface of the tissue section analyzed.
- E-F Immunohistochemistry. Proliferation of the viable tumor component and the average micro-vessel density were determined by Ki67 (D) and CD31 (E) staining, respectively. Results are expressed as an average ⁇ SEM.
- FIG. 2 Association of DT01 with CT significantly decreases the peritoneal tumor volume
- Intrahepatic tumor bearing NMRI NU/NU mice were treated as described in FIG. 4 , and sacrificed 22 days post treatment. The presence of peritoneal metastasis was visually monitored during the duration of treatment and measured at the time of sacrifice.
- A Mean peritoneal tumor volume (mm 3 ) in each treatment group.
- B Representative macroscopic images of peritoneal tumors in each group. Results are expressed as an average ⁇ SEM.
- FIG. 3 Comparison of systemic and local administration in two TNBC xenografted models. NMRI NU/NU mice grafted in the fat pad with TNBC cell lines were treated by SC or IP injections of Dbait during 5 consecutive days for one cycle (left panel) or three cycles (right panel) separated each by two weeks without treatment. Doses of each Dbait injection is indicated in the legend. The Data represent the mean relative tumor volume (Vt/Vi) at the different time after beginning of treatment.
- FIG. 4 DT01 association to carboplatin significantly decreases tumor growth in TNBC model.
- Median tumor volume per treatment group error bars in DT01 treated group and DT01+ carboplatin group indicate the standard error of the mean, SEM).
- FIG. 5 DT01 association to carboplatin significantly increases survival in TNBC model. Kaplan-Meier representation of animal survival in MDA-MB-231 model.
- FIG. 6 Scheme of treatment in FIGS. 4 and 5 .
- Metastatic liver disease from colorectal cancer is a significant clinical problem. This is mainly attributed to non-resectable metastases that frequently display low sensitivities to available chemotherapies and develop drug resistance partly via hyperactivation of some DNA repair functions. Combined therapies have shown some disease control however, there is still a need for more efficient chemotherapies to achieve eradication of CRC liver metastasis.
- Dbait treatment increases sensitivity of HT29 and HCT116 CRC cell lines.
- DT01 the pharmacokinetics, biodistribution and the efficacy of the cholesterol conjugated clinical form of Dbait, DT01, were assessed.
- the chemosensitizing abilities of DT01 were evaluated in association with oxaliplatin and 5-fluorouracil in intrahepatic HT29 xenografted mice used as a model for CRC liver metastasis.
- the high uptake of DT01 indicates that the liver is a specific target.
- the decrease in tumor volume is further associated with significant histological changes in necrosis, proliferation, angiogenesis and apoptosis.
- Repeated cycles of DT01 do not increase chemotherapy toxicity.
- Combining DT01 with conventional chemotherapy may prove to be a safe and effective therapeutic strategy in the treatment of metastatic liver cancer.
- the aims of this study were to firstly demonstrate the efficacy of DT01 in vitro, secondly to assess the pharmacokinetics and the distribution of DT01 in the liver, and thirdly to demonstrate the concomitant impact of systemic DT01 administration in combination with conventional chemotherapy (oxaliplatin with 5′-fluorouracil) in a CRC metastatic liver tumor model.
- Dbait acts by activating DNA-PK kinase, which phosphorylate numerous targets including the histone variant H2AX. They first confirmed the activity of Dbait in two CRC cell lines (HCT116 and HT29) by monitoring the pan-nuclear phosphorylation of H 2AX.
- the inventors determined the number of living cells, at different time points after treatment with Dbait or oxaliplatin (OXA) and 5-fluorouracil (5-FU) or a combination of Dbait with chemotherapy (CT) ( FIG. 1B ).
- OXA oxaliplatin
- 5-fluorouracil 5-fluorouracil
- CT a combination of Dbait with chemotherapy
- HCT116 and HT29 differ by many parameters including their P53 status (HCT116 being proficient whereas HT29 is mutated). In this instance, despite some differences in their sensitivity to standalone Dbait treatment both cell lines were equally sensitive to the combination of CT with Dbait.
- mice were treated with either a single intraperitoneal (IP) or an intravenous (IV) dose of 5 mg of DT01.
- IP administration resulted in a C max of 578 ⁇ g/ml, a T max of 1 hour and an AUC 0-6 of 799 whereas IV led to a C max of 1,917 ⁇ g/ml, a T max of 0.08 hours and an AUC 0-6 of 799.
- Pharmacokinetic analyses revealed that following IP injection, the plasmatic exposure of DT01 was longer than that of IV bolus injection with an AUC corresponding to approximately 70% of the AUC with IV administration.
- the inventors used a fluorescent labelled cy5-DT01 molecule to monitor the biodistribution in excised whole-organs. Both cy5-DT01 and DT01 have similar properties in terms of pharmacokinetics and DNA-PK activation.
- the maximal DT01 fluorescence was observed in the liver, intestines, and the kidneys by both routes with the highest intensities observed in the liver and intestines following IP administration.
- the high fluorescence emitted by the kidneys and urine observed in mice suggest that DT01 is preferentially eliminated by the kidneys.
- there was no measurable DT01 in the blood 6 hours after injection significant amounts of DT01 were still detectable in the liver indicating a specific retention in this organ.
- DT01 activation of DNA-PK in tissue can be revealed by the phosphorylation of the histone H2AX.
- the inventors monitored DT01 activity by analyzing distribution of H2AX phosphorylation in livers bearing HT29 grafted tumors. Interestingly, a high level of ⁇ -H2AX was specifically observed in the tumor and not in the surrounding healthy tissues indicating a preferential uptake or activity of the DT01 molecules in the tumor cells of the liver.
- the inventors used a HT29 xenografted liver tumor model, since previous reports and the in vitro data demonstrate this line to be highly chemo-resistant mainly due to the V600E BRAF mutation.
- the animals were treated with OXA and 5-FU, a treatment close to the traditional FOLFOX protocol for patients, using two different schedules based on biodistribution data ( FIG. 1A ).
- the two schedules consisted of either two or four-hour intervals between the two treatments, since the maximum DT01 levels in the liver were observed at one and three hours after treatment.
- Ki67 immunoreactivity indicated that tumors treated with either DT01 or chemotherapy alone were densely packed with a high degree of proliferation.
- the efficacy was significantly more pronounced at the four-hour time point, in terms of necrosis, apoptosis, proliferation and angiogenesis.
- tumors treated with a delay of four hours between DT01 and CT and sampled 22 days post treatment presented with a proportion of lysed hepatocytes within the tumor and slight edema in the adjacent non-malignant liver, in the absence of further clinical signs of toxicity such as loss of weight. Histological analyses did not reveal morphological signs of toxicity in the other groups ( FIG. 1 ). In addition, liver enzyme tests did not reveal significant differences between the control and the combination treated groups.
- the inventors analyzed the tolerability of DT01 in association with OXA or 5-FU for extended treatment cycles. They determined the toxicity of escalating doses of DT01 (total doses of 30, 50 or 80 mg) following systemic administration for two cycles (5xDT01 administrations per treatment cycle) associated to OXA or 5-FU in a cohort of 50 mice. No loss of weight was observed in animals during or post treatment. Similarly, other clinical signs of toxicity such as diarrhea or behavioral changes were not noted in these mice. At autopsy 6 weeks post the second cycle of treatment, all abdominal organs, the thoracic cavity and contents appeared normal. No major variations in liver weights or histology were observed between the vehicle and combination treated groups.
- DT01 is an attractive drug candidate based on its central role in DNA repair.
- DT01 preferentially accumulate in the liver and intestines after systemic injection. Although the entire liver appeared to be uniformly fluorescent after Cy5-DT01 injection, the activation of DNA-PK revealed by the phosphorylation of H2AX was observed exclusively in tumor cells and not in the healthy tissue surrounding the tumor. This observation indicates that either DT01 does not enter non-tumor cells and/or that DT01 is not active in healthy liver tissue.
- DT01 was specifically designed by cholesterol conjugation firstly, in order to increase the bioavailability and secondly, to play on the difference in the substrate uptake between cancer and normal cells.
- Low density lipoproteins (LDL) are a major component of the cholesterol pathway.
- CRC cell lines HT29 (mutated p53, ATCC: HTB-38) and HCT116 (wild-type p53, ATCC: CCL-247) were purchased directly from ATCC. These cells were authenticated by ATCC by generating human short tandem repeat profiles by simultaneously amplifying multiple STR loci and amelogenin (for gender determination) using the Promega PowerPlex® Systems. These cells were cultured in the laboratory for less than 6 months from the date of purchase in DMEM medium supplemented with 10% fetal bovine serum, 1% sodium pyruvate, 100 mg/ml streptomycin and 100 mg/ml penicillin (Invitrogen, Carlsbad, Calif.), when the current study was performed.
- DMEM medium supplemented with 10% fetal bovine serum, 1% sodium pyruvate, 100 mg/ml streptomycin and 100 mg/ml penicillin (Invitrogen, Carlsbad, Calif.), when the current study was performed.
- HT29 cell line stably expressing luciferase was established in-house using a pGL4.5 luciferase reporter vector (luc2/CMV/Hygro) (Promega).
- HT29 luciferase cells were supplemented with 200 ⁇ g/m1 hygromycin B. All cell lines were additionally subjected to mycoplasma testing in-house and were free of mycoplasma contamination (Biovalley, France).
- Cells were transfected with 2.5 ⁇ gs of Dbait (5′-GCTGTGCCCACAACCCAGCAAACAAGCCTAGA-(H) TCTAGGCTTGTTTGCTGGGTTGTGGGCACAGC-3′ SEQ ID No 9) (Eurogentec, Belgium) where H is a hexaethyleneglycol linker and underlined nucleotides are phosphorothioates.
- the cells were sham transfected with an 8bp oligonucleotide control (8H) complexed with 11 kDa polyethyleneimine (PEI) as previously described (Quantz et al, 2009, PloS one, 4, e6298).
- PEI polyethyleneimine
- ⁇ H2AX immunofluorescence was performed as described previously using a monoclonal anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301 (1:500 dilution; 05-636, Millipore, USA) (9).
- Cells were seeded at a density of 3 ⁇ 10 4 cells/60 mm dishes and transfected with Dbait. Following treatment, cells were washed and left untreated or treated with a combination of 5 ⁇ M of oxaliplatin (OXA, Sigma) and 2.5 ⁇ M of 5-fluouracil (5-FU, Sigma) and live cell counts were performed on days 1, 3, 5, 6, 7 and 9.
- OXA oxaliplatin
- 5-fluouracil 5-FU
- Cells were transfected with Dbait and left untreated or treated with 5 ⁇ M of OXA and 2.5 ⁇ M of 5-FU for 1 hr. The cells were diluted, allowed to grow for 14 days and the clones were stained with crystal violet and counted.
- HT29 Luciferase (HT29L) cells were implanted by direct injection of cell suspensions (1 ⁇ 10 6 /10 ⁇ L of PBS) onto the upper surface of the left lobe. Tumor growth was monitored through bioluminescence analysis (IVIS, Caliper sciences).
- DT01 (Dbait with a cholesterol tetraethylene glycol incorporated at the 5′-end) was used (Agilent technologies, Boulder, Colo.).
- the fluorescent DT01 incorporates a cyanine 5 at the thymidine located immediately after the linker. Six hours after injection, fluorescence imaging was performed using a Typhoon scanner (GE Helathcare).
- DT01 was systemically administered through IP injection at a dose of 5 mgs/day for 5 consecutive days starting on day 0 (DO).
- OXA (6mg/kg, lx per cycle, Day 1)
- 5-FU 25 mg/kg, 3x per cycle, Days 1-3
- mice were sacrificed 22 days post treatment.
- ASAT alkaline phosphatase
- ALP alkaline phosphatase
- GTT glutamyl transpeptidase
- AYL amylase
- TBIL total bilirubin
- OXA and 5′-FU were administered through systemic IP injection at doses of 1 ⁇ 6 mg/kg or 3 ⁇ 25mg/kg, 4 hours after DT01 treatment respectively. Animals were observed regularly for any adverse effects.
- Hematoxylin, eosin, and saffron (HES) stained tumor sections were assessed by an experienced pathologist (Dr. Huerre, Institut Curie) in a blinded fashion. Viable and necrotic components (indicated by increased cell size, indistinct cell border, eosinophilic cytoplasm, loss or condensation of the nucleus, or associated inflammation) were expressed as a proportion (%) of the total tumor surface. Apoptosis was estimated (weak- ⁇ 5%, moderate 5-10%, significant 10-20% and very significant 20-50%) from representative non-necrotic fields at high power.
- Digitization and image capture was performed using a whole-slide scanning system (Philips digital pathology solutions).
- Immunohistochemistry was performed using rabbit anti-Ki67 (ab28364, 1/500; Abcam, UK) and rabbit anti-CD31 (ab15580, 1/500; Abcam, UK) antibodies. This was followed by a secondary biotinylated goat anti-rabbit IgG antibody (BA-1000; Vector, USA) and revealed using a rabbit specific HRP/DAB (ABC) detection kit. Images were captured using a fluorescence microscope (Eclipse 90i, Nikon). The average Ki67 index was scored by establishing a ratio between Ki67 +ve and -ve cells, in five randomly selected microscopic fields per section. Average micro-vessel density was determined by CD31 staining. CD31 positive vessels were counted in five randomly selected microscopic fields per section.
- the objective of the present study was to demonstrate a systemic effect of DT01 alone in a model of breast cancer, in particular of triple negative breast cancer.
- the animal model is mice after 45 days of engraftment. Mice were subcutaneously grafted in mammary fat pad with MDA-MB-231 tumor cells.
- DT01 could control effectively tumor growth in all tested triple negative breast cancer models (mice engrafted with BC227, BC173, MDA-MB-468 and MDA-MB-231 cell lines) by local administration.
- the inventors compared intraperitoneal administration to local administration (intratumoral and peri-tumoral subcutaneous) in two TNBC xenografted models. They surprisingly observed that 3-5 fold more DT01 are required for similar efficacy than local administration.
- the route of administration was intraperitoneal administration which mimics in mouse intravenous perfusion administration in human.
- the dose level of DT01 was 5 mg/animal/day.
- the DT01 intraperitoneal administration was performed during 3 sessions of 5 consecutive days with one week without treatment between each cycle. 13 mice were included, 7 of which receiving DT01.
- the control group received vehicle alone 0.9% NaCl.
- Intraperitoneal DT01 administration is well tolerated.
- DT01 treatment showed a significant better tumor growth control and animal survival than the control group.
- the MDA-MB-231 triple negative breast cancer model was chosen because it was the most resistant to DT01 treatment in previous experiments using intratumoral and peri-tumor subcutaneous administrations.
- each treatment cycle comprised of 5 consecutive days of administration with DT01 at a dose level of 5 mg/animal/day.
- Treatment was administered in 3 cycles with a 2 week gap between cycles either alone or in association with carboplatin. Only one dose of carboplatin was injected per cycle of DT01 treatment (2 nd day of each cycle). The dose of carboplatin was 50 mg/kg per cycle. Treatments were performed over 7 weeks (3 cycles of treatment).
- Intraperitoneal DT01 administration is well tolerated.
- Antitumor activity was evaluated by measuring tumor volume during and after treatment.
- DT01 was administered intraperitoneally during 3 sessions of 5 days treatment with two weeks of rest between each session.
- Carboplatin was administered once a week on the second day of each DT01 treatment cycle.
- DT01+ carboplatin combination treatment showed a better tumor growth control compared to DT01 standalone treatment ( FIG. 4 ). The curve was discontinued on the day of the first death in each group. In addition, the DT01+ carboplatin combination also increases the survival.
- DT01 the cholesterol tetraethylene glycol incorporated form of Dbait was synthesized by automated solid-phase oligonucleotide synthesis (Agilent technologies, USA).
- the MDA-MB231 cell line is derived from a human breast adenocarcinoma and can be ordered at the ATCC.
- the MDA-MB231 cells were grafted in the mammary fat pad with 10.10 6 cells re-suspended in 0.1 ml of DMEM with no additive.
- the athymic nude mouse is immunodeficient, thus enabling the xenotransplantation and growth of human tumors.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Inorganic Chemistry (AREA)
- Oncology (AREA)
- Dermatology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Saccharide Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
Description
- This application is continuation of U.S. application Ser. No. 16/081,045, filed Aug. 30, 2018, which is the U.S. national stage application of International Patent Application No. PCT/EP2017/054702, filed Mar. 1, 2017.
- The Sequence Listing for this application is labeled “Seq-List.txt” which was created on Sep. 13, 2018 and is 7 KB. The entire content of the sequence listing is incorporated herein by reference in its entirety.
- The present invention relates to the field of oncology.
- Breast cancer is the most common female malignancy, with more than 1.7 million new cases diagnosed each year worldwide (Torre, Siegel, Ward, & Jemal, 2015). Molecular classification divides breast cancer in three major sub-groups: LuminalA/B, HER2+ and Basal-like/Triple negative breast cancer (TNBC)(Vuong, Simpson, Green, Cummings, & Lakhani, 2014). Triple-negative breast cancer (TN BC) is an aggressive histological subtype with limited treatment options and very poor prognosis following progression after standard chemotherapeutic regimens. Resistance to current standard therapies such as anthracyclines or taxanes limits the available options for previously treated patients with metastatic TNBC to a small number of non-cross-resistant regimens, and there is currently no preferred standard chemotherapy. Platinum-based regimens are an emerging option for patients with TNBC with BRCA1 mutation.
- Chemo-resistance presents a major obstacle to the efficacy of cancer treatment. DNA repair plays a key role in chemo-resistance by eliminating the damage induced on chromosomes by the chemotherapeutic agents and inhibitors of DNA repair pathways may provide novel opportunities for restoring tumor sensitivity to these treatments.
- Dbait molecules are a new class of DNA repair inhibitors triggering false DNA damage signaling in cancer cells. These molecules are short double-stranded DNA with a free double strand blunt end, which target key damage signal transducers such as DNA dependent protein kinase (DNA-PK) and Poly-ADP-Ribo-Polymerase, triggering their activation and amplifying false damage signaling. Consequently, the recruitment of downstream DNA repair enzymes is impaired, inhibiting several DNA repair pathways such as homologous recombination, non-homologous end joining, base excision repair and single-strand break repair leading to an accumulation of unrepaired damage causing cell death.
- Dbait molecules have been shown to be effective in combination with radiotherapy on several radio-resistant tumors, both in vitro and in vivo. In order to increase the efficiency of cellular uptake, the Dbait molecule was modified by covalently linking a cholesterol moiety to the 5′-end (DT01) (WO2011/161075; Berthault et al, 2011, Cancer gene therapy, 18, 695-706). It has been demonstrated that local administration of DT01 by intra-tumoral injection in association with radiotherapy, increases survival of xenografted human melanoma models (Biau et al, 2014, Neoplasia, 16, 835-844). However to date, the efficacy of systemic administration of DT01, alone or in association with chemotherapy, has not been investigated.
- However, it has been shown that DT01 as an adjunct treatment enhances the therapeutic efficacy of transarterial chemoembolization (TACE) in a rabbit VX2 liver tumor model (Devun et al, Journal of Vascular and Interventional Radiology, 2013, 24, 1080) but TACE derives its beneficial effect by two primary mechanisms. Firstly, most tumors within the liver are supplied by the proper hepatic artery, so arterial embolization preferentially interrupts the tumor's blood supply and stalls growth until neovascularization. Secondly and most importantly, focused administration of chemotherapy allows for delivery of a higher dose to the tissue while simultaneously reducing systemic exposure, which is typically the dose limiting factor.
- Moreover, the molecule DT01 was designed for being used in combination with an endosomolytic agent such as the chloroquine. The chloroquine facilitates the release of coDbait from endosomes into the cytosol and is described as necessary. It was thus shown that coDbait administered by subcutaneous and intratumoral injection in the presence of chloroquine and that concomitant chloroquine treatment was added to the protocol to increase coDbait uptake and efficacy resulting in an increase of the radiosensitivity of xenografted tumors by coDbait (Schlegel et al, 2012, Molecular Therapy—Nucleic Acids, 1, e33).
- It should be further noted that some cancers, especially radioresistant or chemoresistant cancers such as triple-negative breast cancer (TNBC), remain difficult to treat and any improvement in their treatments is important.
- Surprisingly, the inventors observed that the DBait molecules, especially those called coDBait which are conjugated to cholesterol, can be used efficiently for treating cancer, even resistant cancer, by systemic administration, especially intraperitoneal and intravenous administration, without any quinoline endosomlytic agent, especially chloroquine. Indeed, the same efficacy by these administration routes can be obtained with a dose which is only 2-5 fold higher and conjugated DBait has no toxicity. The efficacy of the conjugated DBait with intraperitoneal and intravenous administration has been shown with resistant cancers.
- Therefore, the present invention relates to a nucleic acid molecule for use for treating cancer, wherein the nucleic acid molecule has one of the following formula:
- wherein N is a deoxynucleotide, n is an integer from 15 to 195, the underlined N refers to a nucleotide having or not a modified phosphodiester backbone, L′ is a linker, C is the molecule facilitating endocytosis selected from a lipophilic molecule or a ligand which targets cell receptor enabling receptor mediated endocytosis, L is a linker, m is an integer being 0 or 1 and p is 1;
- wherein the nucleic acid is to be used without combined administration of an endosomolytic agent;
- wherein the nucleic acid is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes.
- Preferably, the nucleic acid of formula (I) has one or several of the following features:
-
- N is a deoxynucleotide selected from the group consisting of A (adenine), C (cytosine), T (thymine) and G (guanine) and selected so as to avoid occurrence of a CpG dinucleotide and to have less than 80% sequence identity to any gene in a human genome; and/or,
- the linked L′ is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane; and/or,
- m is 1 and L is a carboxamido polyethylene glycol, more preferably carboxamido triethylene or tetraethylene glycol; and/or,
- C is selected from the group consisting of a cholesterol, single or double chain fatty acids such as octadecyl, oleic acid, dioleoyl or stearic acid, or ligand (including peptide, protein, aptamer) which targets cell receptor such as folic acid, tocopherol, sugar such as galactose and mannose and their oligosaccharide, peptide such as RGD and bombesin, and protein such transferring and integrin, preferably is a cholesterol or a tocopherol, still more preferably a cholesterol.
- More preferably, the nucleic acid molecule has one of the following formulae:
- wherein the underlined nucleotide refers to a nucleotide having or not a phosphorothioate or methylphosphonate backbone, the linked L′ is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane; m is 1 and L is a carboxamido oligoethylene glycol, C is selected from the group consisting of dioleoyl, octadecyl, folic acid, tocopherol and cholesterol;
- wherein the nucleic acid is to be used without any combined administration of an endosomolytic agent, especially chloroquine; and
- wherein the nucleic acid is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes.
- Still more preferably, the nucleic acid is
- wherein the underlined nucleotide refers to a nucleotide having a phosphorothioate backbone.
- Preferably, the nucleic acid is to be administered by intravenous route, for instance administered by injection, intravenous drip, bolus or pump.
- Preferably, the cancer is a radioresistant or chemoresistant cancer. More preferably, it is selected from the group consisting of a triple-negative breast cancer (TNBC), a chemoresistant hepatocellular carcinoma (HCC), a chemoresistant ovarian cancer, a chemoresistant lung cancer, and a metastatic liver cancer. In a particular embodiment, the cancer is selected from the group consisting of a doxorubicin-resistant hepatocarcinoma (HCC), a platinum-resistant ovarian cancer, a platinum-resistant triple-negative breast cancer and a colorectal liver metastasis.
- Preferably, the nucleic acid is to be used in combination with radiotherapy and/or chemotherapy. In one embodiment, the nucleic acid is to be used in combination with a DNA damaging agent. Preferably, the DNA damaging agent is selected from the group consisting of an inhibitor of topoisomerases I or II, a DNA crosslinker, a DNA alkylating agent, an anti-metabolic agent and inhibitors of the mitotic spindles. More preferably, the nucleic acid is to be used in combination with a chemotherapy selected from the group consisting of doxorubicin, oxaliplatin, carboplatin, cisplatin and 5-FU.
- The present invention relates to a nucleic acid molecule (coDBait) for use for treating cancer, wherein the nucleic acid molecule has one of the following formula:
- wherein N is a deoxynucleotide, n is an integer from 1 to 195, the underlined N refers to a nucleotide having or not a modified phosphodiester backbone, L′ is a linker, C is the molecule facilitating endocytosis selected from a lipophilic molecule or a ligand which targets cell receptor enabling receptor mediated endocytosis, L is a linker, m is an integer being 0 or 1 and p is 1;
- wherein the nucleic acid is to be used without combined administration of an endosomolytic agent;
- wherein the nucleic acid is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes.
- When the absence of a quinoline endosomolytic agent is mentioned herein, it refers to the endosomolytic agent as defined WO2011/161075 in pages 26-28 of (incorporated herein by reference). In particular, the quinoline endosomolytic agent is the chloroquine. In particular, it means that the nucleic acid as described herein is not for simultaneous, separate or sequential use with any endosomolytic agent.
- The present invention relates to
-
- a nucleic acid molecule as defined herein or a pharmaceutical composition comprising it, and optionally a pharmaceutically acceptable carrier, in particular for use in the treatment of cancer without combined administration of an quinoline endosomolytic agent and wherein the pharmaceutical composition is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes, optionally in combination with radiotherapy and/or a DNA-damaging anti-tumor agent;
- the use of a nucleic acid molecule as defined herein or a pharmaceutical composition comprising it for the manufacture of a medicament for treating cancer, wherein the medicament is not used in combination with any quinoline endosomolytic agent and is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes, optionally in combination with radiotherapy and/or a DNA-damaging anti-tumor agent;
- a method for treating cancer in a subject, comprising administering a therapeutic effective amount of a nucleic acid molecule as defined herein or a pharmaceutical composition comprising it by a parenteral systemic route selected from intraperitoneal and intravenous routes but without any administration of a quinoline endosomolytic agent;
- a pharmaceutical composition comprising a nucleic acid molecule as defined herein, a DNA-damaging anti-tumor agent, and a pharmaceutically acceptable carrier, in particular for use in the treatment of cancer without combined administration of an quinoline endosomolytic agent and wherein the pharmaceutical composition is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes;
- a pharmaceutical composition comprising a nucleic acid molecule as defined herein and a DNA-damaging anti-tumor agent for the manufacture of a medicament for treating cancer, wherein the medicament is not used in combination with any quinoline endosomolytic agent and is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes;
- a method for treating cancer in a subject, comprising administering a therapeutic effective amount of a pharmaceutical composition comprising a nucleic acid molecule as defined herein and a DNA-damaging anti-tumor agent by a parenteral systemic route selected from intraperitoneal and intravenous routes but without any administration of a quinoline endosomolytic agent, optionally in combination with radiotherapy and/or a DNA-damaging anti-tumor agent;
- a product or kit containing (a) a nucleic acid molecule as defined herein, and optionally b) a DNA-damaging anti-tumor agent, as a combined preparation for simultaneous, separate or sequential use, in particular in the treatment of cancer without combined administration of an quinoline endosomolytic agent and wherein the pharmaceutical composition is to be administered by a parenteral systemic route selected from intraperitoneal and intravenous routes;
- a method for treating a cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition comprising a nucleic acid molecule as defined herein by a parenteral systemic route selected from intraperitoneal and intravenous routes but without any administration of a quinoline endosomolytic agent, and an effective amount of a pharmaceutical composition comprising a DNA-damaging anti-tumoral agent.
- The terms “kit”, “product” or “combined preparation”, as used herein, defines especially a “kit of parts” in the sense that the combination partners as defined above can be dosed independently or by use of different fixed combinations with distinguished amounts of the combination partners, i.e. simultaneously or at different time points. The parts of the kit of parts can then, e.g., be administered simultaneously or chronologically staggered, that is at different time points and with equal or different time intervals for any part of the kit of parts. The ratio of the total amounts of the combination partners to be administered in the combined preparation can be varied. The combination partners can be administered by the same route or by different routes.
- Within the context of the invention, the term treatment denotes curative, symptomatic, and preventive treatment. Pharmaceutical compositions, kits, products and combined preparations of the invention can be used in humans with existing cancer or tumor, including at early or late stages of progression of the cancer. The pharmaceutical compositions, kits, products and combined preparations of the invention will not necessarily cure the patient who has the cancer but will delay or slow the progression or prevent further progression of the disease, ameliorating thereby the patients' condition. In particular, the pharmaceutical compositions, kits, products and combined preparations of the invention reduce the development of tumors, reduce tumor burden, produce tumor regression in a mammalian host and/or prevent metastasis occurrence and cancer relapse. In treating the cancer, the pharmaceutical composition of the invention is administered in a therapeutically effective amount.
- By “therapeutically effective amount” it is meant the quantity of the pharmaceutical composition of the invention which prevents, removes or reduces the deleterious effects of cancer in mammals, including humans, alone or in combination with the other active ingredients of the pharmaceutical composition, kit, product or combined preparation. It is understood that the administered dose may be lower for each compound in the composition to the “therapeutic effective amount” define for each compound used alone or in combination with other treatments than the combination described here. The “therapeutic effective amount” of the composition will be adapted by those skilled in the art according to the patient, the pathology, the mode of administration, etc.
- Whenever within this whole specification “treatment of a cancer” or the like is mentioned with reference to the pharmaceutical composition of the invention, there is meant: a) a method for treating a cancer, said method comprising administering a pharmaceutical composition of the invention to a subject in need of such treatment; b) the use of a pharmaceutical composition of the invention for the treatment of a cancer; c) the use of a pharmaceutical composition of the invention for the manufacture of a medicament for the treatment of a cancer; and/or d) a pharmaceutical composition of the invention for use in the treatment a cancer.
- The pharmaceutical compositions contemplated herein may include a pharmaceutically acceptable carrier in addition to the active ingredient(s). The term “pharmaceutically acceptable carrier” is meant to encompass any carrier (e.g., support, substance, solvent, etc.) which does not interfere with effectiveness of the biological activity of the active ingredient(s) and that is not toxic to the host to which it is administered. For example, for parental administration, the active compounds(s) may be formulated in a unit dosage form for injection in vehicles such as saline, dextrose solution, serum albumin and Ringer's solution.
- The pharmaceutical composition can be formulated as solutions in pharmaceutically compatible solvents or as emulsions, suspensions or dispersions in suitable pharmaceutical solvents or vehicle in a way known in the art. Formulations suitable for parental administration conveniently comprise a sterile oily or aqueous preparation of the active ingredient which is preferably isotonic with the blood of the recipient. Every such formulation can also contain other pharmaceutically compatible and nontoxic auxiliary agents, such as, e.g. stabilizers, antioxidants, binders, dyes, emulsifiers or flavouring substances. The formulations of the present invention comprise an active ingredient in association with a pharmaceutically acceptable carrier therefore and optionally other therapeutic ingredients. The carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient thereof. The pharmaceutical compositions are advantageously applied by injection or intravenous infusion of suitable sterile solutions. Methods for the safe and effective administration of most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in the standard literature.
- Conjugated DBait Molecules, Called coDBait
- The DBait molecules for use in the present invention can be described by the following formulae:
- wherein N is a nucleotide, n is an integer of at least 1, the underlined N refers to a nucleotide having or not a modified phosphodiester backbone, L′ is a linker, C is a molecule facilitating endocytosis, L is a linker, m is an integer being 0 or 1 and p is 1. Preferably, the underlined N refers to a nucleotide having a modified phosphodiester backbone.
- In preferred embodiments, the molecule of formula (I) has one or several of the following features:
-
- N is a deoxynucleotide, preferably selected from the group consisting of A (adenine), C (cytosine), T (thymine) and G (guanine) and selected so as to avoid occurrence of a CpG dinucleotide and to have less than 80% or 70%, even less than 60% or 50% sequence identity to any gene in a human genome; and/or,
- n is an integer from 15 to 195, from 15 to 95, from 19 to95, from 21 to 95, from 27 to 95, from 15 to 45, from 19 to 45, from 21 to 45, or from 27 to 45. In a particularly preferred embodiment, n is 27; and/or,
- the underlined N refers to a nucleotide having or not a phosphorothioate or methylphosphonate backbone, more preferably a phosphorothioate backbone; preferably, the underlined N refers to a nucleotide having a modified phosphodiester backbone; and/or,
- the linked L′ is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane; and/or,
- m is 1 and L is a carboxamido polyethylene glycol, more preferably carboxamido triethylene or tetraethylene glycol; and/or,
- C is selected from the group consisting of a cholesterol, single or double chain fatty acids such as octadecyl, oleic acid, dioleoyl or stearic acid, or ligand (including peptide, protein, aptamer) which targets cell receptor such as folic acid, tocopherol, sugar such as galactose and mannose and their oligosaccharide, peptide such as RGD and bombesin, and protein such transferring and integrin, preferably is a cholesterol or a tocopherol, still more preferably a cholesterol.
- Preferably, C-Lm is a triethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-triethyleneglycol radical). Alternatively, C-Lm is a tetraethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-tetraethyleneglycol radical).
- In a particular embodiment, the nucleic acid molecules can be Dbait molecules such as those extensively described in PCT patent applications WO2005/040378, WO2008/034866 and WO2008/084087, the disclosure of which is incorporated herein by reference.
- Dbait molecules may be defined by a number of characteristics necessary for their therapeutic activity, such as their minimal length, the presence of at least one free end, and the presence of a double stranded portion, preferably a DNA double stranded portion. As will be discussed below, it is important to note that the precise nucleotide sequence of Dbait molecules does not impact on their activity. Furthermore, Dbait molecules may contain a modified and/or non-natural backbone.
- Preferably, Dbait molecules are of non-human origin (i.e., their nucleotide sequence and/or conformation (e.g., hairpin) does not exist as such in a human cell), most preferably of synthetic origin. As the sequence of the Dbait molecules plays little, if any, role, Dbait molecules have preferably no significant degree of sequence homology or identity to known genes, promoters, enhancers, 5′- or 3′-upstream sequences, exons, introns, and the like. In other words, Dbait molecules have less than 80% or 70%, even less than 60% or 50% sequence identity to any gene in a human genome. Methods of determining sequence identity are well known in the art and include, e.g., Blast. Dbait molecules do not hybridize, under stringent conditions, with human genomic DNA. Typical stringent conditions are such that they allow the discrimination of fully complementary nucleic acids from partially complementary nucleic acids.
- In addition, the sequence of the Dbait molecules is preferably devoid of CpG in order to avoid the well-known toll-like receptor-mediated immunological reactions.
- The length of Dbait molecules may be variable, as long as it is sufficient to allow appropriate binding of Ku protein complex comprising Ku and DNA-PKcs proteins. It has been showed that the length of Dbait molecules must be greater than 20 bp, preferably about 32 bp, to ensure binding to such a Ku complex and allowing DNA-PKcs activation. Preferably, Dbait molecules comprise between 20-200 bp, more preferably 24-100 bp, still more preferably 26-100, and most preferably between 24-200, 25-200, 26-200, 27-200, 28-200, 30-200, 32-200, 24-100, 25-100, 26-100, 27-100, 28-100, 30-100, 32-200 or 32-100 bp. For instance, Dbait molecules comprise between 24-160, 26-150, 28-140, 28-200, 30-120, 32-200 or 32-100 bp. By “bp” is intended that the molecule comprise a double stranded portion of the indicated length.
- In a particular embodiment, the Dbait molecules having a double stranded portion of at least 32 pb, or of about 32 bp, comprise the same nucleotide sequence than Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5). Optionally, the Dbait molecules have the same nucleotide composition than Dbait32, Dbait32Ha, Dbait32Hb, Dbait32Hc or Dbait32Hd but their nucleotide sequence is different. Then, the Dbait molecules comprise one strand of the double stranded portion with 3 A, 6 C, 12 G and 11 T. Preferably, the sequence of the Dbait molecules does not contain any CpG dinucleotide.
- Alternatively, the double stranded portion comprises at least 16, 18, 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5). In a more particular embodiment, the double stranded portion consists in 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- The nucleic acid as disclosed herein must have at least one free end, as a mimic of DSB.
- In a particular embodiment, they contain only one free end. Preferably, Dbait molecules are made of hairpin nucleic acids with a double-stranded DNA stem and a loop. The loop can be a nucleic acid, or other chemical groups known by skilled person or a mixture thereof. A nucleotide linker may include from 2 to 10 nucleotides, preferably, 3, 4 or 5 nucleotides. Non-nucleotide linkers non exhaustively include abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, or other polymeric compounds (e. g. oligoethylene glycols such as those having between 2 and 10 ethylene glycol units, preferably 4, 5, 6, 7 or 8 ethylene glycol units). A preferred linker is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and other linkers such as 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane. Accordingly, in a particular embodiment, the Dbait molecules can be a hairpin molecule having a double stranded portion or stem comprising at least 16, 18, 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5) and a loop being a hexaethyleneglycol linker, a tetradeoxythymidylate linker (T4) or 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane. In a more particular embodiment, those Dbait molecules can have a double stranded portion consisting in 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5).
- Dbait molecules preferably comprise a 2′-deoxynucleotide backbone, and optionally comprise one or several (2, 3, 4, 5 or 6) modified nucleotides and/or nucleobases other than adenine, cytosine, guanine and thymine. Accordingly, the Dbait molecules are essentially a DNA structure. In particular, the double-strand portion or stem of the Dbait molecules is made of deoxyribonucleotides.
- Preferred Dbait molecules comprise one or several chemically modified nucleotide(s) or group(s) at the end of one or of each strand, in particular in order to protect them from degradation. In a particular preferred embodiment, the free end(s) of the Dbait molecules is(are) protected by one, two or three modified phosphodiester backbones at the end of one or of each strand. Preferred chemical groups, in particular the modified phosphodiester backbone, comprise phosphorothioates. Alternatively, preferred Dbait have 3′- 3′ nucleotide linkage, or nucleotides with methylphosphonate backbone. Other modified backbones are well known in the art and comprise phosphoramidates, morpholino nucleic acid, 2′-0,4′-C methylene/ethylene bridged locked nucleic acid, peptide nucleic acid (PNA), and short chain alkyl, or cycloalkyl intersugar linkages or short chain heteroatomic or heterocyclic intrasugar linkages of variable length, or any modified nucleotides known by skilled person. In a first preferred embodiment, the Dbait molecules have the free end(s) protected by one, two or three modified phosphodiester backbones at the end of one or of each strand, more preferably by three modified phosphodiester backbones (in particular phosphorothioate or methylphosphonate) at least at the 3′end, but still more preferably at both 5′ and 3′ ends.
- In a most preferred embodiment, the Dbait molecule is a hairpin nucleic acid molecule comprising a DNA double-stranded portion or stem of 32 bp (e.g., with a sequence selected from the group consisting of SEQ ID Nos 1-5, in particular SEQ ID No 4) and a loop linking the two strands of the DNA double-stranded portion or stem comprising or consisting of a linker selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane, the free ends of the DNA double-stranded portion or stem (i.e. at the opposite of the loop) having three modified phosphodiester backbones (in particular phosphorothioate internucleotidic links).
- Said nucleic acid molecules are made by chemical synthesis, semi-biosynthesis or biosynthesis, any method of amplification, followed by any extraction and preparation methods and any chemical modification. Linkers are provided so as to be incorporable by standard nucleic acid chemical synthesis.
- More preferably, nucleic acid molecules are manufactured by specially designed convergent synthesis: two complementary strands are prepared by standard nucleic acid chemical synthesis with the incorporation of appropriate linker precursor, after their purification, they are covalently coupled together.
- The molecules facilitating endocytosis are conjugated to Dbait molecules, preferably through a linker. Any linker known in the art may be used to covalently attach the molecule facilitating endocytosis to Dbait molecules For instance, WO09/126933 provides a broad review of convenient linkers pages 38-45. The linker can be non-exhaustively, aliphatic chain, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, or other polymeric compounds (e. g. oligoethylene glycols such as those having between 2 and 10 ethylene glycol units, preferably 3, 4, 5, 6, 7 or 8 ethylene glycol units, still more preferably 6 ethylene glycol units), as well as incorporating any bonds that may be break down by chemical or enzymatical way, such as a disulfide linkage, a protected disulfide linkage, an acid labile linkage (e.g., hydrazone linkage), an ester linkage, an ortho ester linkage, a phosphonamide linkage, a biocleavable peptide linkage, an azo linkage or an aldehyde linkage. Such cleavable linkers are detailed in WO2007/040469 pages 12-14, in WO2008/022309 pages 22-28.
- In a particular embodiment, the nucleic acid molecule can be linked to one molecule facilitating endocytosis. Alternatively, several molecules facilitating endocytosis (e.g., two, three or four) can be attached to one nucleic acid molecule.
- In a specific embodiment, the linker between the molecule facilitating endocytosis, in particular cholesterol, and nucleic acid molecule is CO—NH—(CH2—CH2—O)n, wherein n is an integer from 1 to 10, preferably n being selected from the group consisting of 3, 4, 5 and 6. In a very particular embodiment, the linker is CO—NH—(CH2—CH2—O)4 (carboxamido tetraethylene glycol) or CO—NH—(CH2—CH2—O)3 (carboxamido triethylene glycol). The linker can be linked to nucleic acid molecules at any convenient position which does not modify the activity of the nucleic acid molecules. In particular, the linker can be linked at the 5′ end. Therefore, in a preferred embodiment, the contemplated conjugated Dbait molecule is a Dbait molecule having a hairpin structure and being conjugated to the molecule facilitating endocytosis, preferably through a linker, at its 5′ end.
- In another specific embodiment, the linker between the molecule facilitating endocytosis, in particular cholesterol, and nucleic acid molecule is dialkyl-disulfide {e.g., (CH2)r—S—S—(CH2)s with r and s being integer from 1 to 10, preferably from 3 to 8, for instance 6}.
- In a most preferred embodiment, the conjugated Dbait molecule is a hairpin nucleic acid molecule comprising a DNA double-stranded portion or stem of 32 bp (e.g., with a sequence selected from the group consisting of SEQ ID Nos 1-5, in particular SEQ ID No 4) and a loop linking the two strands of the DNA double-stranded portion or stem comprising or consisting of a linker selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane, the free ends of the DNA double-stranded portion or stem (i.e. at the opposite of the loop) having three modified phosphodiester backbones (in particular phosphorothioate internucleotidic links) and said Dbait molecule being conjugated to a cholesterol at its 5′ end, preferably through a linker (e.g. carboxamido oligoethylene glycol, preferably carboxamido triethylene or tetraethylene glycol).
- In a preferred embodiment, NNNN—(N)n—N comprises at least 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb
- (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5) or consists in 20, 22, 24, 26, 28, 30 or 32 consecutive nucleotides of Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5). In a particular embodiment, NNNN—(N)n—N comprises or consists in Dbait32 (SEQ ID No 1), Dbait32Ha (SEQ ID No 2), Dbait32Hb (SEQ ID No 3), Dbait32Hc (SEQ ID No 4) or Dbait32Hd (SEQ ID No 5), more preferably Dbait32Hc (SEQ ID No 4).
- According, the conjugated Dbait molecule or hairpin nucleic acid molecule may be selected from the group consisting of:
- with the same definition than formulae (I) for L, L′, C, p and m.
- In preferred embodiments, the molecule of formulae (Ia), (Ib), (Ic), (Id), and (Ie), has one or several of the following features:
-
- the underlined nucleotide refers to a nucleotide having or not a phosphorothioate or methylphosphonate backbone, more preferably a phosphorothioate backbone; preferably, the underlined nucleotide refers to a nucleotide having a phosphorothioate or methylphosphonate backbone, more preferably a phosphorothioate backbone; and/or,
- the linked L′ is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane; and/or,
- m is 1 and L is a carboxamido polyethylene glycol, more preferably carboxamido triethylene or tetraethylene glycol; and/or,
- p is 1; and/or,
- C is selected from the group consisting of a cholesterol, single or double chain fatty acids such as octadecyl, oleic acid, dioleoyl or stearic acid, or ligand (including peptide, protein, aptamer) which targets cell receptor such as folic acid, tocopherol, sugar such as galactose and mannose and their oligosaccharide, peptide such as RGD and bombesin, and protein such transferring and integrin, preferably is a cholesterol.
- Preferably, C-Lm is a triethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-triethyleneglycol radical) or a tetraethyleneglycol linker (10-)-[1-propyl-3-N-carbamoylcholesteryl]-tetraethyleneglycol radical).
- In a specific embodiment of the Dbait molecules or hairpin nucleic acid molecules of formulae (I), (Ia), (Ib), (Ic), (Id), and (Ie), L′ is preferably selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.
- In a specific embodiment of the Dbait molecules or hairpin nucleic acid molecules of formulae (I), (Ia), (Ib), (Ic), (Id), and (Ie), with C being cholesterol, C-Lm is the radical
- In a preferred embodiment, the conjugated Dbait molecule or hairpin nucleic acid molecule is selected from the group consisting of (I), (Ia), (Ib), (Ic), (Id), and (Ie), wherein C-Lm being the radical
- and wherein L′ is preferably selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4) and 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane, more preferably 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.
- In a very specific embodiment, the Dbait molecule or hairpin nucleic acid molecule has the following formula
- wherein C-Lm is a tetraethyleneglycol linker (10-O-[1-propyl-3-N-carbamoylcholesteryl]-tetraethyleneglycol radical, and L′ is 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane and wherein the underlined nucleotides have a phosphorothioate backbone.
- Accordingly, the molecule has the following structure and it is referred thereto in the Example section as “coDbait” or “DT01”.
- Another representation of this molecule is shown below:
- An alternative molecule is the following:
- In another preferred embodiment, the nucleic acid molecule has one of the following formulae
- wherein N is a deoxynucleotide, n is an integer from 1 to 195, the underlined N refers to a nucleotide having or not a modified phosphodiester backbone, L′ is a linker, C is a cholesterol, L is a linker, m is an integer being 0 or 1, and p is 1. Preferably, the underlined N refers to a nucleotide having a modified phosphodiester backbone.
- DNA Damaging Treatment
- In addition to the conjugated Dbait molecules, the treatment may also further comprise an antitumor treatment, preferably a treatment by a DNA damaging agent or radiotherapy. The DNA-damaging treatment can be radiotherapy or chemotherapy with a DNA-damaging antitumor agent, or a combination thereof.
- DNA strand breakage can be achieved by ionized radiation (radiotherapy). Radiotherapy includes, but is not limited to, γ-rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells. Other radiotherapies include microwaves and UV-irradiation. Other approaches to radiation therapy are also contemplated in the present invention.
- The DNA-damaging antitumor agent is preferably selected from the group consisting of an inhibitor of topoisomerases I or II, a DNA crosslinker, a DNA alkylating agent, an anti-metabolic agent and inhibitors of the mitotic spindles.
- Inhibitors of topoisomerases I and/or II include, but are not limited to, etoposide, topotecan, camptothecin, irinotecan, amsacrine, intoplicine, anthracyclines such as doxorubicin, epirubicine, daunorubicine, idanrubicine and mitoxantrone. Inhibitors of Topoisomerase I and II include, but are not limited to, intoplecin. In a preferred embodiment, the DNA-damaging antitumor agent is doxorubicin.
- DNA crosslinkers include, but are not limited to, cisplatin, carboplatin and oxaliplatin. In a preferred embodiment, the DNA-damaging antitumor agent is selected from the group consisting of carboplatin and oxaliplatin.
- Anti-metabolic agents block the enzymes responsible for nucleic acid synthesis or become incorporated into DNA, which produces an incorrect genetic code and leads to apoptosis. Non-exhaustive examples thereof include, without limitation, folic acid antagonists, pyrimidine analogs, purine analogs and adenosine deaminase inhibitors, and more particularly Methotrexate, Floxuridine, Cytarabine, 6-Mercaptopurine, 6- Thioguanine, Fludarabine phosphate, Pentostatine, 5-fluorouracil (5-FU), gemcitabine and capecitabine.
- The DNA-damaging anti-tumor agent can be alkylating agents including, without limitation, nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, metal salts and triazenes. Non-exhaustive examples thereof include Uracil mustard, Chlormethine, Cyclophosphamide (CYTOXAN(R)), Ifosfamide, Melphalan, Chlorambucil, Pipobroman, Triethylenemelamine, Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine, Fotemustine, cisplatin, carboplatin, oxaliplatin, thiotepa, Streptozocin, Dacarbazine, and Temozolomide.
- Inhibitors of the mitotic spindles include, but are not limited to, paclitaxel, docetaxel, vinorelbine, larotaxel (also called XRP9881; Sanofi-Aventis), XRP6258 (Sanofi-Aventis), BMS-184476 (Bristol-Meyer-Squibb), BMS-188797 (Bristol-Meyer-Squibb), BMS-275183 (Bristol-Meyer-Squibb), ortataxel (also called IDN 5109, BAY 59-8862 or SB-T-101131 ; Bristol-Meyer-Squibb), RPR 109881A (Bristol-Meyer-Squibb), RPR 116258 (Bristol-Meyer-Squibb), NBT-287 (TAPESTRY), PG-paclitaxel (also called CT-2103, PPX, paclitaxel poliglumex, paclitaxel polyglutamate or Xyotax™), ABRAXANE® (also called Nab-Paclitaxel; ABRAXIS BIOSCIENCE), Tesetaxel (also called DJ-927), IDN 5390 (INDENA), Taxoprexin (also called docosahexanoic acid-paclitaxel ; PROTARGA), DHA-paclitaxel (also called Taxoprexin®), and MAC-321 (WYETH). Also see the review of Hennenfent & Govindan (2006, Annals of Oncology, 17, 735-749).
- Preferably, the DNA-damaging antitumor agent is an inhibitor of topoisomerases I and/or II, a DNA crosslinker, an anti-metabolic agent or a combination thereof. In a preferred embodiment, the DNA-damaging antitumor agent is selected from the group consisting of doxorubicin, 5-FU, carboplatin and oxaliplatin or a combination thereof. In a most preferred embodiment, the conjugated DBait is DT01 and the DNA-damaging antitumor agent is selected from the group consisting of doxorubicin, carboplatin, 5-FU and oxaliplatin.
- Cancers or Tumors to be Treated
- The pharmaceutical compositions and the products, kits or combined preparation described in the invention can be used for treating cancer in a subject.
- The terms “cancer”, “cancerous”, or “malignant” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, for example, leukemia, lymphoma, blastoma, carcinoma and sarcoma. More particular examples of such cancers include chronic myeloid leukemia, acute lymphoblastic leukemia, Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ALL), squamous cell carcinoma, small-cell lung cancer, non-small cell lung cancer, glioma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon carcinoma, and head and neck cancer, gastric cancer, germ cell tumor, pediatric sarcoma, sinonasal natural killer, multiple myeloma, acute myelogenous leukemia (AML), chronic lymphocytic leukemia, mastocytosis and any symptom associated with mastocytosis.
- “Leukemia” refers to progressive, malignant diseases of the blood-forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease—acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number of abnormal cells in the blood—leukemic or aleukemic (subleukemic). Leukemia includes, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocyte leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblasts leukemia, myelocytic leukemia, myeloid granulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, plasmacytic leukemia, promyelocytic leukemia, Rieder cell leukemia, Schilling's leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia. In certain aspects, the present invention provides treatment for chronic myeloid leukemia, acute lymphoblastic leukemia, and/or Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ALL).
- Various cancers are also encompassed by the scope of the invention, including, but not limited to, the following: carcinoma including that of the bladder (including accelerated and metastatic bladder cancer), breast, colon (including colorectal cancer), kidney, liver, lung (including small and non-small cell lung cancer and lung adenocarcinoma), ovary, prostate, testes, genitourinary tract, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), esophagus, stomach, gall bladder, cervix, thyroid, and skin (including squamous cell carcinoma); hematopoietic tumors of lymphoid lineage including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, histiocytic lymphoma, and Burketts lymphoma; hematopoietic tumors of myeloid lineage including acute and chronic myelogenous leukemias, myelodysplastic syndrome, myeloid leukemia, and promyelocytic leukemia; tumors of the central and peripheral nervous system including astrocytoma, neuroblastoma, glioma, and schwannomas; tumors of mesenchymal origin including fibrosarcoma, rhabdomyosarcoma, and osteosarcoma; other tumors including melanoma, xenoderma pigmentosum, keratoactanthoma, seminoma, thyroid follicular cancer, and teratocarcinoma; melanoma, unresectable stage III or IV malignant melanoma, squamous cell carcinoma, small-cell lung cancer, non-small cell lung cancer, glioma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon carcinoma, and head and neck cancer, retinoblastoma, gastric cancer, germ cell tumor, bone cancer, bone tumors, adult malignant fibrous histiocytoma of bone; childhood malignant fibrous histiocytoma of bone, sarcoma, pediatric sarcoma, sinonasal natural killer, neoplasms, plasma cell neoplasm; myelodysplastic syndromes; neuroblastoma; testicular germ cell tumor, intraocular melanoma, myelodysplastic syndromes; myelodysplastic/myeloproliferative diseases, synovial sarcoma. In addition, disorders include urticaria pigmentosa, mastocytosises such as diffuse cutaneous mastocytosis, solitary mastocytoma in human, as well as dog mastocytoma and some rare subtypes like bullous, erythrodermic and teleangiectatic mastocytosis, mastocytosis with an associated hematological disorder, such as a myeloproliferative or myelodysplastic syndrome, or acute leukemia, myeloproliferative disorder associated with mastocytosis, mast cell leukemia, in addition to other cancers. Other cancers are also included within the scope of disorders including, but are not limited to, the following: carcinoma, including that of the bladder, urothelial carcinoma, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid, testis, particularly testicular seminomas, and skin; including squamous cell carcinoma; gastrointestinal stromal tumors (“GIST”); hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma and Burketts lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; other tumors, including melanoma, seminoma, teratocarcinoma, neuroblastoma and glioma; tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma, and schwannomas; tumors of mesenchymal origin, including fibrosarcoma, rhabdomyosarcoma, and osteosarcoma; and other tumors, including melanoma, xenoderma pigmentosum, keratoactanthoma, seminoma, thyroid follicular cancer, teratocarcinoma, chemotherapy refractory non-seminomatous germ-cell tumors, and Kaposi's sarcoma, and any metastasis thereof.
- In a preferred embodiment of the present invention, the cancer is a solid tumor. The term “solid tumor” especially means breast cancer, ovarian cancer, cancer of the colon and generally the GI (gastro-intestinal) tract, cervix cancer, lung cancer, in particular small- cell lung cancer, and non-small-cell lung cancer, head and neck cancer, bladder cancer, cancer of the prostate or Kaposi's sarcoma.
- The pharmaceutical compositions and the products, kits or combined preparations described in the invention may be useful for inhibiting the growth of solid tumors, decreasing the tumor volume, preventing the metastatic spread of tumors and the growth or development of micrometastases. The pharmaceutical compositions and the products, kits or combined preparations described in the invention are in particular suitable for the treatment of poor prognosis patients or of radio- or chemo-resistant tumors.
- In one embodiment, the cancer can be selected from melanoma, glioblastoma, breast cancer, colon cancer, lung cancer, gastrointestinal cancer, liver cancer and head and neck cancer.
- In a preferred embodiment, the cancer is a radioresistant or chemoresistant cancer. More particularly, the cancer is selected from the group consisting of a radioresistant melanoma, a triple-negative breast cancer (TNBC), a chemoresistant hepatocellular carcinoma (HCC), a chemoresistant lung cancer, a chemoresistant ovarian cancer and a metastatic liver cancer. More specifically, the cancer is selected from the group consisting of a doxorubicin-resistant hepatocarcinoma (HCC), a platinum-resistant triple-negative breast cancer, a platinum-resistant ovarian cancer and a colorectal liver metastasis.
- In a specific embodiment, the present invention relates to the use of conjugated DBait molecules for the treatment of colorectal carcinoma in combination with oxaliplatin and 5-FU. Preferably, the colorectal carcinoma is metastatic, more preferably with metastasis in the liver and/or peritoneum.
- In a very specific embodiment, the present invention relates to the use of conjugated DBait molecules for the treatment of radioresistant or chemoresistant cancer which is not localized into the abdominal cavity. Such cancers do not encompass colorectal carcinoma (e.g. CRC metastasis of the liver and the peritoneum) accordingly. More particularly, the cancer is selected from the group consisting of a TNBC and a chemoresistant ovarian cancer. More specifically, the cancer is selected from the group consisting of a platinum-resistant TNBC and a platinum-resistant ovarian cancer.
- In a further embodiment, the present invention relates to the use of conjugated DBait molecules for the treatment of triple negative breast cancer. More specifically, the conjugated DBait molecules is used in combination with platinum-containing anti-cancer drugs, especially selected from cisplatin, oxaliplatin and carboplatin. In a specific embodiment, the present invention relates to the use of conjugated DBait molecules for the treatment of triple-negative breast cancer (TN BC) in combination with carboplatin. Optionally, the treatment can be combined with radiotherapy.
- Regimen, Dosages and Administration Routes
- The effective dosage of each of the combination partners employed in the combined preparation of the invention may vary depending on the particular compound or pharmaceutical composition employed, the mode of administration, the condition being treated, the severity of the condition being treated. Thus, the dosage regimen of the combined preparation of the invention is selected in accordance with a variety of factors including the route of administration and the patient status. A physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the single active ingredients required to prevent, counter or arrest the progress of the condition. Optimal precision in achieving concentration of the active ingredients within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the active ingredients' availability to target sites.
- When a DNA-damaging antitumor agent is used in combination with the conjugated Dbait molecule, the DNA-damaging antitumor agent and the conjugated Dbait molecules may be administered by the same route or by distinct routes. The administration route for the DNA-damaging antitumor agent may be oral, parenteral, intravenous, intratumoral, subcutaneous, intracranial, intraartery, topical, rectal, transdermal, intradermal, nasal, intramuscular, intraosseous, and the like.
- The conjugated Dbait molecules is to be administered before and/or simultaneously with and/or after the irradiation and/or the administration of the DNA-damaging antitumor agent, more preferably before and/or simultaneously with the irradiation and/or the administration of the DNA-damaging antitumor agent. The irradiation and/or the administration of the DNA-damaging antitumor agent is performed so as the conjugated Dbait molecules are present in the tumoral cells when the irradiation is applied or when the DNA-damaging antitumor agent reach the tumoral cells. The physician, clinician or veterinarian of ordinary skill can determine the regimen based on the active ingredients, their kinetics of availability to target sites or their pharmacokinetic profiles in plasma. Preliminary results indicate that conjugated Dbait molecules stay active during one day. In a first preferred embodiment, the irradiation is to be applied or the DNA-damaging antitumor agent is to be administered at the beginning of the treatment with conjugated Dbait molecules or after the treatment with conjugated Dbait molecules. For instance, the irradiation is to be applied or the DNA-damaging antitumor agent is to be administered 3-24 h after the beginning of the treatment with conjugated Dbait molecules. The DNA-damaging antitumor agent and conjugated Dbait molecules may also be simultaneously administered.
- Once the treatment by radiotherapy or with the DNA-damaging antitumor agent has begun, the treatment with the conjugated Dbait molecules can continue as long as the treatment by radiotherapy or with the DNA-damaging antitumor agent is to be applied or administered. Alternatively, the treatment with the conjugated Dbait molecules can also end.
- For conjugated Dbait molecules, the effective dosage of the DNA-damaging antitumor agent employed in the combined preparation, kit or product of the invention may vary depending on the mode of administration, the condition being treated, the severity of the condition being treated. Thus, the dosage regimen of the conjugated Dbait molecules is selected in accordance with a variety of factors including the route of administration and the patient status. A physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the conjugated Dbait molecules required to prevent, counter or arrest the progress of the cancer, in particular in combination with the selected DNA damaging treatment.
- The one skilled in the art can adapt the amount in order to obtain an efficient amount of the conjugated Dbait molecules in the tumor of at least 0.01 mg per 1 cm3 of tumor, preferably 0.1 - 40 mg per 1 cm3 of tumor, most preferably 1 - 20 mg per 1 cm3 of tumor, in particular in a daily treatment protocol or in a weekly treatment protocol. For instance, for an intravenous or intraperitoneal route, the efficient amount or unit dosage of the conjugated Dbait molecules may be of 0.1 to 100 mg, preferably of 4 to 40 mg. Accordingly, the efficient amount or unit dosage of the conjugated Dbait molecules may be of 0.06 to 0.6 mg/kg of patient. Of course, the dosage and the regimen can be adapted by the one skilled in art in consideration of the chemotherapy and/or radiotherapy regimen.
- For radiotherapy, any radiotherapy regimen known in the art may be used, in particular stereotactic irradiation (e.g., 15 Gy) or a fractionated irradiation. The use of a fractionated irradiation may be particularly efficient, for instance irradiation may applied every day or every 2-5 days, preferably every 3-4 days, in a period of one, two, three, four, five or six weeks. The irradiation may be from 1 to 10 Gy, preferably from 2 to 5 Gy, in particular 2, 3, 4 or 5 Gy. For instance, fractionated irradiation of 15×2Gy in six weeks, or of 4 to 6×5Gy in two weeks may be contemplated. In a preferred embodiment, the contemplated radiotherapy is a protocol with 4 irradiations of 5 Gy in two weeks. Different regimens or conditions of combined treatments of cancer with irradiation and Dbait molecules have been tested and allowed to demonstrate the radio-sensibilization of tumors by Dbait molecules depends on the doses of Dbait molecules but not of the irradiation doses.
- For chemotherapy, the effective dosage of the DNA-damaging antitumor agent employed in the combined preparation, kit or product of the invention or in combination with the composition of the invention may vary depending on the particular DNA-damaging antitumor agent employed, the mode of administration, the condition being treated, the severity of the condition being treated. Thus, the dosage regimen of the DNA-damaging antitumor agent is selected in accordance with a variety of factors including the route of administration and the patient status. A physician, clinician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the DNA-damaging antitumor agent required to prevent, counter or arrest the progress of the cancer.
- The treatment may include one or several cycles, for instance two to ten cycles, in particular two, three, four or five cycles. The cycles may be continued or separated. For instance, each cycle is separated by a period of time of one to eight weeks, preferably three to four weeks.
- Further aspects and advantages of the present invention will be disclosed in the following experimental section, which should be regarded as illustrative and not limiting the scope of the present application. A number of references are cited in the present specification; each of these cited references is incorporated herein by reference.
-
FIG. 1 : DT01 significantly increases sensitivity to CT in a CRC (HT29) liver metastatic model Intrahepatic tumor bearing NMRINU/NU mice were treated as described in the Material and Methods, and sacrificed 22 days post treatment. Livers were sampled for macroscopic and microscopic examination. (A) Sequence of DT01 and CT therapy. CT was administered 2 or 4 hours post DT01 treatment. (B) Mean liver tumor volume (mm3) in each treatment group. (C) Representative macroscopic and HES sections of liver tumors in each group. (D) Tumor necrotic component assessed through HES staining. Necrosis is expressed as a proportion (%) of the total tumor surface of the tissue section analyzed. (E-F) Immunohistochemistry. Proliferation of the viable tumor component and the average micro-vessel density were determined by Ki67 (D) and CD31 (E) staining, respectively. Results are expressed as an average±SEM. -
FIG. 2 : Association of DT01 with CT significantly decreases the peritoneal tumor volume Intrahepatic tumor bearing NMRINU/NU mice were treated as described inFIG. 4 , and sacrificed 22 days post treatment. The presence of peritoneal metastasis was visually monitored during the duration of treatment and measured at the time of sacrifice. (A) Mean peritoneal tumor volume (mm3) in each treatment group. (B) Representative macroscopic images of peritoneal tumors in each group. Results are expressed as an average±SEM. -
FIG. 3 : Comparison of systemic and local administration in two TNBC xenografted models. NMRINU/NU mice grafted in the fat pad with TNBC cell lines were treated by SC or IP injections of Dbait during 5 consecutive days for one cycle (left panel) or three cycles (right panel) separated each by two weeks without treatment. Doses of each Dbait injection is indicated in the legend. The Data represent the mean relative tumor volume (Vt/Vi) at the different time after beginning of treatment. -
FIG. 4 : DT01 association to carboplatin significantly decreases tumor growth in TNBC model. Median tumor volume per treatment group (error bars in DT01 treated group and DT01+ carboplatin group indicate the standard error of the mean, SEM). -
FIG. 5 : DT01 association to carboplatin significantly increases survival in TNBC model. Kaplan-Meier representation of animal survival in MDA-MB-231 model. -
FIG. 6 : Scheme of treatment inFIGS. 4 and 5 . - Metastatic liver disease from colorectal cancer (CRC) is a significant clinical problem. This is mainly attributed to non-resectable metastases that frequently display low sensitivities to available chemotherapies and develop drug resistance partly via hyperactivation of some DNA repair functions. Combined therapies have shown some disease control however, there is still a need for more efficient chemotherapies to achieve eradication of CRC liver metastasis.
- The inventors investigated the tolerance and efficacy of Dbait in association with conventional chemotherapy. In vitro, Dbait treatment increases sensitivity of HT29 and HCT116 CRC cell lines. In vivo, the pharmacokinetics, biodistribution and the efficacy of the cholesterol conjugated clinical form of Dbait, DT01, were assessed. The chemosensitizing abilities of DT01 were evaluated in association with oxaliplatin and 5-fluorouracil in intrahepatic HT29 xenografted mice used as a model for CRC liver metastasis. The high uptake of DT01 indicates that the liver is a specific target. The inventors demonstrated significant anti-tumor efficacy in a liver metastasis model with DT01 treatment in combination with oxaliplatin and 5-fluorouracil (mean: 501 vs 872 mm2, p=0.02) compared to chemotherapy alone. The decrease in tumor volume is further associated with significant histological changes in necrosis, proliferation, angiogenesis and apoptosis. Repeated cycles of DT01 do not increase chemotherapy toxicity. Combining DT01 with conventional chemotherapy may prove to be a safe and effective therapeutic strategy in the treatment of metastatic liver cancer.
- The aims of this study were to firstly demonstrate the efficacy of DT01 in vitro, secondly to assess the pharmacokinetics and the distribution of DT01 in the liver, and thirdly to demonstrate the concomitant impact of systemic DT01 administration in combination with conventional chemotherapy (oxaliplatin with 5′-fluorouracil) in a CRC metastatic liver tumor model.
- Results
- Dbait Treatment Increases Sensitivity of Colon Cancer Cell Lines to Chemotherapy
- The inventors have previously shown that Dbait acts by activating DNA-PK kinase, which phosphorylate numerous targets including the histone variant H2AX. They first confirmed the activity of Dbait in two CRC cell lines (HCT116 and HT29) by monitoring the pan-nuclear phosphorylation of H 2AX.
- To first investigate the effects of Dbait on cell survival to chemotherapy, the inventors determined the number of living cells, at different time points after treatment with Dbait or oxaliplatin (OXA) and 5-fluorouracil (5-FU) or a combination of Dbait with chemotherapy (CT) (
FIG. 1B ). As already observed in fibroblasts (Quanz, 2009, PloS one, 4, e6298), Dbait alone appears to have no effect on cell proliferation in both cell lines. Treatment with OXA and 5-FU resulted in a decrease in cell proliferation. However, the level of proliferation was significantly reduced by day 9 in cells transfected with Dbait prior to chemotherapy treatment in both HCT116 and HT29 cell lines compared to chemotherapy alone (p<0.001 and p<0.02, respectively). These differences become apparent particularly at later time points (>5 days after treatment) indicating that the increase of efficacy with Dbait may be a slow process. - To confirm the chemosensitization effect of Dbait in combination with OXA and 5-FU, clonogenic survival assays were performed on HCT116 and HT29. HCT116 cells showed approximately 30% (p<0.01) lethality after Dbait treatment alone revealing their dependency in repair activity for survival whereas no significant effect was noted in HT29. Since the sensitivity of HCT116 to Dbait was not detected during the first 8 days of proliferation, this result suggests that the cells growing with Dbait accumulate lethal lesions that impair their survival later on. Treatment with chemotherapy alone (OXA/5-FU) resulted in a significant decrease in the survival of HCT116 (p<0.001) whereas only a trend was observed with the HT29 cell line (p=0.08). However, combination of Dbait with chemotherapy resulted in a significant reduction in survival in both cell lines (p=0.05). HCT116 and HT29 differ by many parameters including their P53 status (HCT116 being proficient whereas HT29 is mutated). In this instance, despite some differences in their sensitivity to standalone Dbait treatment both cell lines were equally sensitive to the combination of CT with Dbait.
- Pharmacokinetic and Biodistribution Analyses of Intraperitoneal Vs Intravenous Administration of DT01
- To avoid transfectant adjuvant toxicity, all in vivo studies were performed with DT01, a Dbait-cholesterol conjugate facilitating the cellular uptake of these molecules without added toxicity. To determine the best route for systemic administration of DT01 mice were treated with either a single intraperitoneal (IP) or an intravenous (IV) dose of 5 mg of DT01. IP administration resulted in a Cmax of 578μg/ml, a Tmax of 1 hour and an AUC0-6 of 799 whereas IV led to a Cmax of 1,917 μg/ml, a Tmax of 0.08 hours and an AUC0-6 of 799. Pharmacokinetic analyses revealed that following IP injection, the plasmatic exposure of DT01 was longer than that of IV bolus injection with an AUC corresponding to approximately 70% of the AUC with IV administration.
- The inventors used a fluorescent labelled cy5-DT01 molecule to monitor the biodistribution in excised whole-organs. Both cy5-DT01 and DT01 have similar properties in terms of pharmacokinetics and DNA-PK activation. The maximal DT01 fluorescence was observed in the liver, intestines, and the kidneys by both routes with the highest intensities observed in the liver and intestines following IP administration. The high fluorescence emitted by the kidneys and urine observed in mice suggest that DT01 is preferentially eliminated by the kidneys. Although there was no measurable DT01 in the
blood 6 hours after injection, significant amounts of DT01 were still detectable in the liver indicating a specific retention in this organ. - As already demonstrated in vitro, DT01 activation of DNA-PK in tissue can be revealed by the phosphorylation of the histone H2AX. The inventors monitored DT01 activity by analyzing distribution of H2AX phosphorylation in livers bearing HT29 grafted tumors. Interestingly, a high level of γ-H2AX was specifically observed in the tumor and not in the surrounding healthy tissues indicating a preferential uptake or activity of the DT01 molecules in the tumor cells of the liver.
- DT01 Significantly Increases Sensitivity to OXA and 5-FU In Vivo
- To explore the interest of associating DT01 with the frontline treatment for metastatic CRC, the inventors used a HT29 xenografted liver tumor model, since previous reports and the in vitro data demonstrate this line to be highly chemo-resistant mainly due to the V600E BRAF mutation. The animals were treated with OXA and 5-FU, a treatment close to the traditional FOLFOX protocol for patients, using two different schedules based on biodistribution data (
FIG. 1A ). The two schedules consisted of either two or four-hour intervals between the two treatments, since the maximum DT01 levels in the liver were observed at one and three hours after treatment. - As previously observed in vitro, the tumors were highly resistant to CT alone and DT01 had only a moderate effect when administered alone (
FIGS. 1B, 1C ). Interestingly, the association of DT01 to OXA and 5-FU significantly decreased the liver tumor size in both combination treated groups compared to CT alone when administered at two (mean volume: 525.80 vs 872.01 mm2, p=0.03) and four hours (mean volume: 501.05 vs 872.01 mm2, p=0.02) before CT (FIGS. 1B, 1C ). This effect was not observed when DT01 was associated with a single chemotherapy agent, either OXA or 5-FU. Detailed blinded histological analyses including measures of the viable tumor area, necrosis and apoptosis were assessed in haematoxylin-eosin-saffron (HES) stained sections, by an experienced pathologist. Both groups with DT01 and CT combined treatment showed higher treatment efficacy than the groups receiving single treatment, with a marked increase in necrosis in the group treated with a four-hour interval between CT and DT01 (p<0.0001) than two hours (p<0.01), compared to CT alone (FIG. 1D ). Furthermore, a high apoptotic index was apparent in both groups treated with DT01 and CT. Similar to other histological parameters, the extent of apoptosis was elevated in animals treated with a four-hour delay (p<0.0001). These histological findings were not apparent in the DT01 or chemotherapy alone treated groups. - For many solid tumors, proliferation and microvascularization are indispensable prerequisites for tumor development and metastasis. To further investigate these parameters, immunostaining for Ki67 and CD31, markers of cell proliferation and angiogenesis respectively, were performed in the viable tumor component (
FIGS. 1E, 1F ). Ki67 immunoreactivity indicated that tumors treated with either DT01 or chemotherapy alone were densely packed with a high degree of proliferation. - Treatment with a two-hour interval between DT01 and CT resulted in a moderate decrease in proliferating cells (p=0.02) (
FIG. 1E ). Strikingly, immunoreactivity of Ki67 was 10-fold reduced in the group treated with a four-hour interval between DT01 and CT (p<0.001) (FIG. 1E ). In this group, immunoreactivity was detected only in the tumor rim due to the high degree of necrosis observed in the center core region of the tumor. In addition, diminished intratumoral vessel densities were detected in groups treated with a combination of DT01 and CT, compared to CT alone (FIG. 1F ). However, the mean microvessel density was even more notably reduced in the group treated with a four-hour interval (p<0.001) compared to two hours (p=0.02). Despite similarities in the anti-tumor effect on tumor growth at both the two and four-hour treatment schedules, histologically the efficacy was significantly more pronounced at the four-hour time point, in terms of necrosis, apoptosis, proliferation and angiogenesis. - Unexpectedly, tumors treated with a delay of four hours between DT01 and CT and sampled 22 days post treatment presented with a proportion of lysed hepatocytes within the tumor and slight edema in the adjacent non-malignant liver, in the absence of further clinical signs of toxicity such as loss of weight. Histological analyses did not reveal morphological signs of toxicity in the other groups (
FIG. 1 ). In addition, liver enzyme tests did not reveal significant differences between the control and the combination treated groups. - Interestingly, no further edema was observed when animals receiving the same treatment were sacrificed between 30-65 days. This suggests that the edema observed at
day 22, is reversible over time. Despite the significant tumor efficacy observed 22 days post treatment, tumors monitored after this time point resumed progression. Histological analysis revealed that the proliferative component reached ˜50% at 30-45 days post treatment, only slightly below the level observed in non-treated tumors. - To confirm that combination treatment did not induce additional toxicity to the liver, the inventors analyzed the tolerability of DT01 in association with OXA or 5-FU for extended treatment cycles. They determined the toxicity of escalating doses of DT01 (total doses of 30, 50 or 80 mg) following systemic administration for two cycles (5xDT01 administrations per treatment cycle) associated to OXA or 5-FU in a cohort of 50 mice. No loss of weight was observed in animals during or post treatment. Similarly, other clinical signs of toxicity such as diarrhea or behavioral changes were not noted in these mice. At
autopsy 6 weeks post the second cycle of treatment, all abdominal organs, the thoracic cavity and contents appeared normal. No major variations in liver weights or histology were observed between the vehicle and combination treated groups. - These results suggest that the reversible edema detected after combined treatment in animals bearing hepatic tumor is likely an acute reaction to the tumor response to efficient combination treatment.
- Peritoneal Metastasis Treatment
- CRC often metastasizes to the liver and the peritoneum. Interestingly 90% of the mice intrahepatically xenografted with CRC tumors developed peritoneal metastasis. This property allowed the inventors to monitor the effect of DT01 not only on liver tumors but also on peritoneal metastasis. Animals receiving a combination of DT01 and chemotherapy displayed significantly decreased peritoneal tumor volumes when compared to chemotherapy alone at both the two (mean volume: 300.31 vs 867.20 mm2, respectively, p<0.01) and four hour time intervals (mean volume: 259.51 vs 867.20 mm2, respectively, p<0.01) (
FIG. 2A, 2B ). Although a slight decrease in tumor volume was observed in the group treated with DT01 alone, this did not reach statistical significance. - Discussion
- Approximately 50% of patients with CRC will present either with liver and/or peritoneal metastases or develop them throughout the course of their disease. A majority of patients with CRC hepatic metastases present with non-resectable disease and systemic chemotherapy represents the main if not the only form of therapy. However, the therapeutic window of chemotherapy is limited due to tumor resistance and high toxicity to non-targeted tissue. In such clinical situations, an aggressive chemotherapy regimen alone may not only fail to improve survival, but may also adversely affect the quality of life. Consequently the mortality of these patients remains high. Therefore development of new agents' specifically targeting DNA repair to circumvent chemoresistance and sparing healthy tissues is imperative in the treatment of these cancers. DT01 is an attractive drug candidate based on its central role in DNA repair.
- In the present study, the inventors showed for the first time that systemic DT01 treatment sensitizes CRC cells to conventional chemotherapies by in vitro and in vivo assays. In a CRC metastatic model, they demonstrated significant anti-tumor efficacy in the liver and the peritoneum (regarded as a terminal condition) with DT01 treatment in combination with OXA and 5-FU. It is of interest to note, that the significant anti-tumor effect was limited to DT01 association with both OXA and 5-FU and not with single agent chemotherapy. This demonstrates that in agreement with the clinical conventional setting, combination with DT01 must be associated to double chemotherapy rather than single-agent chemotherapy in the treatment of CRC metastases. This study further highlights that tumors receiving double chemotherapy combined with DT01 restart proliferation and re-growth at later time points (post 22 days). Therefore repeated cycles of treatment would be necessary to achieve long term disease control similar to current conventional chemotherapy protocols. This would be possible as no added toxicity was observed with DT01 alone or in combination with OXA or 5-FU.
- DT01 preferentially accumulate in the liver and intestines after systemic injection. Although the entire liver appeared to be uniformly fluorescent after Cy5-DT01 injection, the activation of DNA-PK revealed by the phosphorylation of H2AX was observed exclusively in tumor cells and not in the healthy tissue surrounding the tumor. This observation indicates that either DT01 does not enter non-tumor cells and/or that DT01 is not active in healthy liver tissue. DT01 was specifically designed by cholesterol conjugation firstly, in order to increase the bioavailability and secondly, to play on the difference in the substrate uptake between cancer and normal cells. Low density lipoproteins (LDL) are a major component of the cholesterol pathway. High requirement for LDL by malignant cells and thus the consequent overexpression of LDL receptors has been shown in many types of cancer cells making tumor cells specific targets of DT01. Additionally, an extensive analysis of normal and cancerous human tissues by immunohistochemistry revealed that either DNA-PKcs or Ku80 were consistently absent in the liver and the mammary epithelium, a specific post-transcriptional regulation that was not found in the other tissues and most of the tumors. Taken together, these data highlight that DT01 is likely to be an efficient drug for the treatment of liver cancers.
- In conclusion, there is an urgent need for new treatment options targeting secondary hepatic malignancies, a rapidly progressive disease with a poor prognosis and an alarming rate of mortality. The present study demonstrated that combining systemic administration of DT01 with conventional chemotherapy can be a safe and effective therapeutic strategy in the treatment of CRC metastasis of the liver and the peritoneum.
- Materials and Methods
- Cell Culture, Constructs, Dbait Molecules, Immunofluorescence and Western Blotting
- CRC cell lines; HT29 (mutated p53, ATCC: HTB-38) and HCT116 (wild-type p53, ATCC: CCL-247) were purchased directly from ATCC. These cells were authenticated by ATCC by generating human short tandem repeat profiles by simultaneously amplifying multiple STR loci and amelogenin (for gender determination) using the Promega PowerPlex® Systems. These cells were cultured in the laboratory for less than 6 months from the date of purchase in DMEM medium supplemented with 10% fetal bovine serum, 1% sodium pyruvate, 100 mg/ml streptomycin and 100 mg/ml penicillin (Invitrogen, Carlsbad, Calif.), when the current study was performed. HT29 cell line stably expressing luciferase was established in-house using a pGL4.5 luciferase reporter vector (luc2/CMV/Hygro) (Promega). HT29 luciferase cells were supplemented with 200μg/m1 hygromycin B. All cell lines were additionally subjected to mycoplasma testing in-house and were free of mycoplasma contamination (Biovalley, France).
- Cells were transfected with 2.5 μgs of Dbait (5′-GCTGTGCCCACAACCCAGCAAACAAGCCTAGA-(H) TCTAGGCTTGTTTGCTGGGTTGTGGGCACAGC-3′ SEQ ID No 9) (Eurogentec, Belgium) where H is a hexaethyleneglycol linker and underlined nucleotides are phosphorothioates. The cells were sham transfected with an 8bp oligonucleotide control (8H) complexed with 11 kDa polyethyleneimine (PEI) as previously described (Quantz et al, 2009, PloS one, 4, e6298).
- γH2AX immunofluorescence was performed as described previously using a monoclonal anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301 (1:500 dilution; 05-636, Millipore, USA) (9).
- In Vitro Proliferation Assay
- Cells were seeded at a density of 3×104 cells/60 mm dishes and transfected with Dbait. Following treatment, cells were washed and left untreated or treated with a combination of 5 μM of oxaliplatin (OXA, Sigma) and 2.5 μM of 5-fluouracil (5-FU, Sigma) and live cell counts were performed on
days - Clonogenic Assay
- Cells were transfected with Dbait and left untreated or treated with 5 μM of OXA and 2.5 μM of 5-FU for 1 hr. The cells were diluted, allowed to grow for 14 days and the clones were stained with crystal violet and counted.
- In Vivo Experiments
- The current study was carried out in strict accordance with the European Union guidelines for animal care. All animal experimentation was approved by the ethics committees of the Institut Curie and the French ministry. Surgical procedures were performed under anesthesia with local analgesia to minimize suffering.
- Animals
- Six week old female NMRINu/Nu mice (Janvier, France) weighing 20-22 g were housed in specific pathogen free environment on a 12 h light and 12 h dark schedule with food and water ad libitum. No more than 6 animals were housed per cage and they were acclimated for at least one week prior to initiating in vivo studies.
- Intrahepatic HT29L Grafting
- HT29 Luciferase (HT29L) cells were implanted by direct injection of cell suspensions (1×106/10 μL of PBS) onto the upper surface of the left lobe. Tumor growth was monitored through bioluminescence analysis (IVIS, Caliper sciences).
- DT01 Molecule
- For in vivo studies, DT01 (Dbait with a cholesterol tetraethylene glycol incorporated at the 5′-end) was used (Agilent technologies, Boulder, Colo.).
- Pharmacokinetics of DT01
- HT29L grafted mice were treated with a single intraperitoneal (IP, n=4) or intravenous (IV, n=3) injection of 5 mg of DT01. Blood samples were harvested prior to treatment and 1, 5, 10, 30 mins, 1 hr, 2 hrs, 4 hrs and 6 hrs post treatment. Plasma was recovered through centrifugation and assayed by ELISA.
- Fluorescence Measurement of Organs
- As the ELISA technique failed to produce reliable quantification in tissues, we used fluorescent imaging, a reliable technique for assessing molecule distribution (15). NMRINU/NU mice were injected with 1 mg of the DT01 fluorescent molecule (DT01-Cy5) through IP (n=3) or IV (n=3) administration.
- The fluorescent DT01 (DT01-Cy5) incorporates a
cyanine 5 at the thymidine located immediately after the linker. Six hours after injection, fluorescence imaging was performed using a Typhoon scanner (GE Helathcare). - DT01 and Chemotherapy Treatment
- HT29L grafted animals (n=49) were allocated into treatment groups and administered one cycle of treatment. DT01 was systemically administered through IP injection at a dose of 5 mgs/day for 5 consecutive days starting on day 0 (DO). OXA (6mg/kg, lx per cycle, Day 1) and 5-FU (25 mg/kg, 3x per cycle, Days 1-3) were administered 2 or 4 hours after DT01 treatment. These mice were sacrificed 22 days post treatment.
- An additional group treated with DT01 and OXA/5-FU at the 4 hour interval (n=10) were kept after treatment until the termination guidelines were met to assess the duration of treatment efficacy.
- Liver Function Assessment
- Blood samples were obtained through submandibular bleeding in lithium heparin tubes (Sarstedt) on
days - (ASAT), alkaline phosphatase (ALP), glutamyl transpeptidase (GGT), amylase (AMYL) and total bilirubin (TBIL) were measured using an MS-Scan II (Melet Schloesing Laboratories, France).
- Toxicity Assays
- NMRINU/NU mice (n=50) were treated with two cycles of DT01 at escalating doses of 3 mg/day (30 mg total), 5 mg/day (50 mg total) or 8 mg/day (80 mg total) through IP injection in combination with OXA or 5′-FU. OXA and 5′-FU were administered through systemic IP injection at doses of 1×6 mg/kg or 3×25mg/kg, 4 hours after DT01 treatment respectively. Animals were observed regularly for any adverse effects.
- Histology
- Hematoxylin, eosin, and saffron (HES) stained tumor sections were assessed by an experienced pathologist (Dr. Huerre, Institut Curie) in a blinded fashion. Viable and necrotic components (indicated by increased cell size, indistinct cell border, eosinophilic cytoplasm, loss or condensation of the nucleus, or associated inflammation) were expressed as a proportion (%) of the total tumor surface. Apoptosis was estimated (weak-<5%, moderate 5-10%, significant 10-20% and very significant 20-50%) from representative non-necrotic fields at high power.
- Digitization and image capture was performed using a whole-slide scanning system (Philips digital pathology solutions).
- Ki67 and CD31 Immunohistochemistry
- Immunohistochemistry was performed using rabbit anti-Ki67 (ab28364, 1/500; Abcam, UK) and rabbit anti-CD31 (ab15580, 1/500; Abcam, UK) antibodies. This was followed by a secondary biotinylated goat anti-rabbit IgG antibody (BA-1000; Vector, USA) and revealed using a rabbit specific HRP/DAB (ABC) detection kit. Images were captured using a fluorescence microscope (Eclipse 90i, Nikon). The average Ki67 index was scored by establishing a ratio between Ki67 +ve and -ve cells, in five randomly selected microscopic fields per section. Average micro-vessel density was determined by CD31 staining. CD31 positive vessels were counted in five randomly selected microscopic fields per section.
- Statistical Analysis
- In vitro experiments were performed with a minimum of two independent experiments. Two-sided unpaired t-tests were used for comparison of cell mortality and survival. Kruskal-Wallis tests were used to compare tumor volumes, and histological data. Error bars indicate standard error of the mean (SEM), except when specifically indicated. All statistical analyses were performed using StatEL software (adScience, France) and a P value of 0.05 was considered statistically significant.
- DT01 Effect Alone on a TNBC Model
- The objective of the present study was to demonstrate a systemic effect of DT01 alone in a model of breast cancer, in particular of triple negative breast cancer. The animal model is mice after 45 days of engraftment. Mice were subcutaneously grafted in mammary fat pad with MDA-MB-231 tumor cells.
- In previous experiments, the inventors demonstrated that DT01 could control effectively tumor growth in all tested triple negative breast cancer models (mice engrafted with BC227, BC173, MDA-MB-468 and MDA-MB-231 cell lines) by local administration.
- As shown in
FIG. 3 , the inventors compared intraperitoneal administration to local administration (intratumoral and peri-tumoral subcutaneous) in two TNBC xenografted models. They surprisingly observed that 3-5 fold more DT01 are required for similar efficacy than local administration. - The route of administration was intraperitoneal administration which mimics in mouse intravenous perfusion administration in human. The dose level of DT01 was 5 mg/animal/day. The DT01 intraperitoneal administration was performed during 3 sessions of 5 consecutive days with one week without treatment between each cycle. 13 mice were included, 7 of which receiving DT01. The control group received vehicle alone 0.9% NaCl.
- No toxicity was observed. Intraperitoneal DT01 administration is well tolerated.
- DT01 treatment showed a significant better tumor growth control and animal survival than the control group.
- The MDA-MB-231 triple negative breast cancer model was chosen because it was the most resistant to DT01 treatment in previous experiments using intratumoral and peri-tumor subcutaneous administrations.
- This experiment confirms that standalone administration of DT01 delays tumor growth in breast cancer tumor.
- Effect of the combination of DT01 with carboplatin on a TNBC model
- As shown in
FIG. 6 , each treatment cycle comprised of 5 consecutive days of administration with DT01 at a dose level of 5 mg/animal/day. Treatment was administered in 3 cycles with a 2 week gap between cycles either alone or in association with carboplatin. Only one dose of carboplatin was injected per cycle of DT01 treatment (2nd day of each cycle). The dose of carboplatin was 50 mg/kg per cycle. Treatments were performed over 7 weeks (3 cycles of treatment). - During the experiment no toxicity is observed. No sign of toxicity such as loss of weight in DT01 treated group. No increase in weight loss or toxicity in DT01+ carboplatin treated group was observed compared to the carboplatin group.
- No abnormal death occurred during the 177 days of the experiment, except one in the carboplatin alone treated group.
- Intraperitoneal DT01 administration is well tolerated.
- Antitumor activity was evaluated by measuring tumor volume during and after treatment. DT01 was administered intraperitoneally during 3 sessions of 5 days treatment with two weeks of rest between each session. Carboplatin was administered once a week on the second day of each DT01 treatment cycle.
- DT01+ carboplatin combination treatment showed a better tumor growth control compared to DT01 standalone treatment (
FIG. 4 ). The curve was discontinued on the day of the first death in each group. In addition, the DT01+ carboplatin combination also increases the survival. - In this study, the treatment of DT01 combining with carboplatin is efficient and leads to a better tumor growth delay than single treatments.
- Materials and Methods
- DT01 Molecule
- DT01, the cholesterol tetraethylene glycol incorporated form of Dbait was synthesized by automated solid-phase oligonucleotide synthesis (Agilent technologies, USA).
- Cells & Animals
- The MDA-MB231 cell line is derived from a human breast adenocarcinoma and can be ordered at the ATCC. The MDA-MB231 cells were grafted in the mammary fat pad with 10.106 cells re-suspended in 0.1 ml of DMEM with no additive. The athymic nude mouse is immunodeficient, thus enabling the xenotransplantation and growth of human tumors.
Claims (13)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/038,704 US20210023116A1 (en) | 2016-03-01 | 2020-09-30 | Treatment of cancer by systemic administration of dbait molecules |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP16305234.3 | 2016-03-01 | ||
EP16305234 | 2016-03-01 | ||
PCT/EP2017/054702 WO2017148976A1 (en) | 2016-03-01 | 2017-03-01 | Treatment of cancer by systemic administration of dbait molecules |
US201816081045A | 2018-08-30 | 2018-08-30 | |
US17/038,704 US20210023116A1 (en) | 2016-03-01 | 2020-09-30 | Treatment of cancer by systemic administration of dbait molecules |
Related Parent Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2017/054702 Continuation WO2017148976A1 (en) | 2016-03-01 | 2017-03-01 | Treatment of cancer by systemic administration of dbait molecules |
US16/081,045 Continuation US10821128B2 (en) | 2016-03-01 | 2017-03-01 | Treatment of cancer by systemic administration of Dbait molecules |
Publications (1)
Publication Number | Publication Date |
---|---|
US20210023116A1 true US20210023116A1 (en) | 2021-01-28 |
Family
ID=55588189
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/081,045 Active 2037-07-28 US10821128B2 (en) | 2016-03-01 | 2017-03-01 | Treatment of cancer by systemic administration of Dbait molecules |
US17/038,704 Abandoned US20210023116A1 (en) | 2016-03-01 | 2020-09-30 | Treatment of cancer by systemic administration of dbait molecules |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/081,045 Active 2037-07-28 US10821128B2 (en) | 2016-03-01 | 2017-03-01 | Treatment of cancer by systemic administration of Dbait molecules |
Country Status (7)
Country | Link |
---|---|
US (2) | US10821128B2 (en) |
EP (1) | EP3423068A1 (en) |
JP (1) | JP6949859B2 (en) |
KR (2) | KR20220127360A (en) |
CN (1) | CN109069528B (en) |
CA (1) | CA3016355A1 (en) |
WO (1) | WO2017148976A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20200407720A1 (en) * | 2018-03-13 | 2020-12-31 | Onxeo | A dbait molecule against acquired resistance in the treatment of cancer |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3461488A1 (en) * | 2017-09-27 | 2019-04-03 | Onxeo | Combination of a dbait molecule and a hdac inhibitor for treating cancer |
JP7450622B2 (en) * | 2018-12-21 | 2024-03-15 | ヴァレリオ・セラピューティクス | Novel conjugated nucleic acid molecules and their uses |
MX2021009863A (en) * | 2019-03-21 | 2021-11-12 | Onxeo | A dbait molecule in combination with kinase inhibitor for the treatment of cancer. |
WO2021148581A1 (en) | 2020-01-22 | 2021-07-29 | Onxeo | Novel dbait molecule and its use |
WO2021178295A1 (en) * | 2020-03-03 | 2021-09-10 | University Of Washington | Synthetic agonists of dna-pk and their use |
TW202210633A (en) * | 2020-06-05 | 2022-03-16 | 法商昂席歐公司 | A dbait molecule in combination with kras inhibitor for the treatment of cancer |
AR122644A1 (en) | 2020-06-19 | 2022-09-28 | Onxeo | NEW CONJUGATED NUCLEIC ACID MOLECULES AND THEIR USES |
CA3240558A1 (en) | 2021-12-16 | 2023-06-22 | Valerio Therapeutics | New conjugated nucleic acid molecules and their uses |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008084087A2 (en) * | 2007-01-12 | 2008-07-17 | Centre National De La Recherche Scientifique | Dbait and its standalone uses thereof |
US7595302B2 (en) * | 2003-10-24 | 2009-09-29 | Institut Curie | Nucleic acids useful for triggering tumor cell lethality |
WO2009126933A2 (en) * | 2008-04-11 | 2009-10-15 | Alnylam Pharmaceuticals, Inc. | Site-specific delivery of nucleic acids by combining targeting ligands with endosomolytic components |
US7741308B2 (en) * | 2003-10-24 | 2010-06-22 | Institut Curie | DBAIT and uses thereof |
US20130131155A1 (en) * | 2010-06-22 | 2013-05-23 | Dna Therapeutics | Optimized In Vivo Delivery System with Endosomolytic Agents for Nucleic Acid Conjugates |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2527440A1 (en) * | 2011-05-27 | 2012-11-28 | Institut Curie | Cancer treatment by combining DNA molecules mimicking double strand breaks with hyperthermia |
-
2017
- 2017-03-01 JP JP2018545659A patent/JP6949859B2/en active Active
- 2017-03-01 CN CN201780014979.XA patent/CN109069528B/en active Active
- 2017-03-01 WO PCT/EP2017/054702 patent/WO2017148976A1/en active Application Filing
- 2017-03-01 KR KR1020227030539A patent/KR20220127360A/en not_active Application Discontinuation
- 2017-03-01 US US16/081,045 patent/US10821128B2/en active Active
- 2017-03-01 CA CA3016355A patent/CA3016355A1/en active Pending
- 2017-03-01 KR KR1020187027868A patent/KR102441432B1/en active IP Right Grant
- 2017-03-01 EP EP17708463.9A patent/EP3423068A1/en active Pending
-
2020
- 2020-09-30 US US17/038,704 patent/US20210023116A1/en not_active Abandoned
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7595302B2 (en) * | 2003-10-24 | 2009-09-29 | Institut Curie | Nucleic acids useful for triggering tumor cell lethality |
US7741308B2 (en) * | 2003-10-24 | 2010-06-22 | Institut Curie | DBAIT and uses thereof |
WO2008084087A2 (en) * | 2007-01-12 | 2008-07-17 | Centre National De La Recherche Scientifique | Dbait and its standalone uses thereof |
WO2009126933A2 (en) * | 2008-04-11 | 2009-10-15 | Alnylam Pharmaceuticals, Inc. | Site-specific delivery of nucleic acids by combining targeting ligands with endosomolytic components |
US20130131155A1 (en) * | 2010-06-22 | 2013-05-23 | Dna Therapeutics | Optimized In Vivo Delivery System with Endosomolytic Agents for Nucleic Acid Conjugates |
US9687557B2 (en) * | 2010-06-22 | 2017-06-27 | Onxeo | Optimized in vivo delivery system with endosomolytic agents for nucleic acid conjugates |
US10363317B2 (en) * | 2010-06-22 | 2019-07-30 | Onxeo | Optimized in vivo delivery system with endosomolytic agents for nucleic acid conjugates |
Non-Patent Citations (1)
Title |
---|
Galluzzi et al. 2015. Autophagy in malignant transformation and cancer progression. EMBO J. 34(7):856-880 (Year: 2015) * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20200407720A1 (en) * | 2018-03-13 | 2020-12-31 | Onxeo | A dbait molecule against acquired resistance in the treatment of cancer |
Also Published As
Publication number | Publication date |
---|---|
US20190091254A1 (en) | 2019-03-28 |
CA3016355A1 (en) | 2017-09-08 |
KR20180118716A (en) | 2018-10-31 |
WO2017148976A1 (en) | 2017-09-08 |
EP3423068A1 (en) | 2019-01-09 |
CN109069528B (en) | 2021-11-05 |
KR20220127360A (en) | 2022-09-19 |
JP2019507763A (en) | 2019-03-22 |
JP6949859B2 (en) | 2021-10-13 |
CN109069528A (en) | 2018-12-21 |
KR102441432B1 (en) | 2022-09-07 |
US10821128B2 (en) | 2020-11-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20210023116A1 (en) | Treatment of cancer by systemic administration of dbait molecules | |
US10765758B2 (en) | Optimized in vivo delivery system with endosomolytic agents for nucleic acid conjugates | |
US9205101B2 (en) | Cancer treatment by combining DNA molecules mimicking double strand breaks with hyperthermia | |
US20230235327A1 (en) | A dbait molecule in combination with kras inhibitor for the treatment of cancer | |
AU2015202211B2 (en) | Optimized in vivo delivery system with endosomolytic agents for nucleic acid conjugates |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: INSTITUT CURIE, FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:DUTREIX, MARIE;BERTHAULT, NATHALIE;REEL/FRAME:053937/0297 Effective date: 20181217 Owner name: CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE, FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:DUTREIX, MARIE;BERTHAULT, NATHALIE;REEL/FRAME:053937/0297 Effective date: 20181217 Owner name: ONXEO, FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:DUTREIX, MARIE;BERTHAULT, NATHALIE;REEL/FRAME:053937/0297 Effective date: 20181217 Owner name: INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE, FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:DUTREIX, MARIE;BERTHAULT, NATHALIE;REEL/FRAME:053937/0297 Effective date: 20181217 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: APPLICATION DISPATCHED FROM PREEXAM, NOT YET DOCKETED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
AS | Assignment |
Owner name: VALERIO THERAPEUTICS, FRANCE Free format text: CHANGE OF NAME;ASSIGNOR:ONXEO;REEL/FRAME:065489/0815 Effective date: 20230606 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |