WO2023014206A1 - 방사선 치료와 병용되는 유전자 치료제로서 암 특이적 트랜스-스플라이싱 리보자임 - Google Patents
방사선 치료와 병용되는 유전자 치료제로서 암 특이적 트랜스-스플라이싱 리보자임 Download PDFInfo
- Publication number
- WO2023014206A1 WO2023014206A1 PCT/KR2022/011774 KR2022011774W WO2023014206A1 WO 2023014206 A1 WO2023014206 A1 WO 2023014206A1 KR 2022011774 W KR2022011774 W KR 2022011774W WO 2023014206 A1 WO2023014206 A1 WO 2023014206A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cancer
- gene
- pharmaceutical composition
- ribozyme
- present
- Prior art date
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 140
- 108090000994 Catalytic RNA Proteins 0.000 title claims abstract description 117
- 201000011510 cancer Diseases 0.000 title claims abstract description 117
- 108091092562 ribozyme Proteins 0.000 title claims abstract description 113
- 102000053642 Catalytic RNA Human genes 0.000 title claims abstract description 112
- 238000001959 radiotherapy Methods 0.000 title claims abstract description 33
- 238000001415 gene therapy Methods 0.000 title abstract description 9
- 230000005855 radiation Effects 0.000 claims abstract description 68
- 230000014509 gene expression Effects 0.000 claims abstract description 56
- 230000001124 posttranscriptional effect Effects 0.000 claims abstract description 7
- 208000016691 refractory malignant neoplasm Diseases 0.000 claims abstract description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 145
- 238000011282 treatment Methods 0.000 claims description 77
- 239000013598 vector Substances 0.000 claims description 58
- 108091007780 MiR-122 Proteins 0.000 claims description 51
- 239000008194 pharmaceutical composition Substances 0.000 claims description 50
- 108091051828 miR-122 stem-loop Proteins 0.000 claims description 43
- 210000001519 tissue Anatomy 0.000 claims description 34
- 108020004999 messenger RNA Proteins 0.000 claims description 32
- 108010017842 Telomerase Proteins 0.000 claims description 24
- 201000007270 liver cancer Diseases 0.000 claims description 24
- 208000014018 liver neoplasm Diseases 0.000 claims description 24
- 239000013604 expression vector Substances 0.000 claims description 20
- 230000008685 targeting Effects 0.000 claims description 19
- 238000001476 gene delivery Methods 0.000 claims description 17
- 241000701022 Cytomegalovirus Species 0.000 claims description 16
- 102100032938 Telomerase reverse transcriptase Human genes 0.000 claims description 14
- 239000003814 drug Substances 0.000 claims description 14
- 229940079593 drug Drugs 0.000 claims description 14
- 239000002773 nucleotide Substances 0.000 claims description 14
- 125000003729 nucleotide group Chemical group 0.000 claims description 14
- 150000007523 nucleic acids Chemical group 0.000 claims description 13
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 10
- 108700026220 vif Genes Proteins 0.000 claims description 10
- 230000006907 apoptotic process Effects 0.000 claims description 9
- 230000001105 regulatory effect Effects 0.000 claims description 9
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 claims description 8
- 102000012406 Carcinoembryonic Antigen Human genes 0.000 claims description 8
- 208000005017 glioblastoma Diseases 0.000 claims description 8
- 230000000295 complement effect Effects 0.000 claims description 7
- 230000002829 reductive effect Effects 0.000 claims description 7
- 101710087044 Cytoskeleton-associated protein 2 Proteins 0.000 claims description 6
- 102100028630 Cytoskeleton-associated protein 2 Human genes 0.000 claims description 6
- 108010072866 Prostate-Specific Antigen Proteins 0.000 claims description 6
- 108700008625 Reporter Genes Proteins 0.000 claims description 6
- 108010026331 alpha-Fetoproteins Proteins 0.000 claims description 6
- 102000013529 alpha-Fetoproteins Human genes 0.000 claims description 6
- 230000000890 antigenic effect Effects 0.000 claims description 6
- 206010006187 Breast cancer Diseases 0.000 claims description 5
- 208000026310 Breast neoplasm Diseases 0.000 claims description 5
- 206010009944 Colon cancer Diseases 0.000 claims description 5
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 5
- 241000700584 Simplexvirus Species 0.000 claims description 5
- 108700025716 Tumor Suppressor Genes Proteins 0.000 claims description 5
- 241001492404 Woodchuck hepatitis virus Species 0.000 claims description 5
- 208000006990 cholangiocarcinoma Diseases 0.000 claims description 5
- 208000019425 cirrhosis of liver Diseases 0.000 claims description 5
- 208000032839 leukemia Diseases 0.000 claims description 5
- 201000005202 lung cancer Diseases 0.000 claims description 5
- 208000020816 lung neoplasm Diseases 0.000 claims description 5
- 201000001441 melanoma Diseases 0.000 claims description 5
- 102000044209 Tumor Suppressor Genes Human genes 0.000 claims description 4
- 230000001772 anti-angiogenic effect Effects 0.000 claims description 4
- 231100000433 cytotoxic Toxicity 0.000 claims description 4
- 230000001472 cytotoxic effect Effects 0.000 claims description 4
- 108020005345 3' Untranslated Regions Proteins 0.000 claims description 3
- 229930195730 Aflatoxin Natural products 0.000 claims description 3
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 claims description 3
- 206010004593 Bile duct cancer Diseases 0.000 claims description 3
- 206010005003 Bladder cancer Diseases 0.000 claims description 3
- 206010005949 Bone cancer Diseases 0.000 claims description 3
- 208000018084 Bone neoplasm Diseases 0.000 claims description 3
- 208000003174 Brain Neoplasms Diseases 0.000 claims description 3
- 206010008909 Chronic Hepatitis Diseases 0.000 claims description 3
- 206010061825 Duodenal neoplasm Diseases 0.000 claims description 3
- 208000000461 Esophageal Neoplasms Diseases 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 206010016654 Fibrosis Diseases 0.000 claims description 3
- 208000022072 Gallbladder Neoplasms Diseases 0.000 claims description 3
- 241000700721 Hepatitis B virus Species 0.000 claims description 3
- 208000008839 Kidney Neoplasms Diseases 0.000 claims description 3
- 206010025323 Lymphomas Diseases 0.000 claims description 3
- 206010030155 Oesophageal carcinoma Diseases 0.000 claims description 3
- 206010033128 Ovarian cancer Diseases 0.000 claims description 3
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 3
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 3
- 206010060862 Prostate cancer Diseases 0.000 claims description 3
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 3
- 206010038389 Renal cancer Diseases 0.000 claims description 3
- 206010039491 Sarcoma Diseases 0.000 claims description 3
- 206010054184 Small intestine carcinoma Diseases 0.000 claims description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 3
- 208000024313 Testicular Neoplasms Diseases 0.000 claims description 3
- 206010057644 Testis cancer Diseases 0.000 claims description 3
- 102000006601 Thymidine Kinase Human genes 0.000 claims description 3
- 108020004440 Thymidine kinase Proteins 0.000 claims description 3
- 208000024770 Thyroid neoplasm Diseases 0.000 claims description 3
- 206010046431 Urethral cancer Diseases 0.000 claims description 3
- 206010046458 Urethral neoplasms Diseases 0.000 claims description 3
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 3
- 208000002495 Uterine Neoplasms Diseases 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000005409 aflatoxin Substances 0.000 claims description 3
- 208000026900 bile duct neoplasm Diseases 0.000 claims description 3
- 230000007882 cirrhosis Effects 0.000 claims description 3
- 201000000312 duodenum cancer Diseases 0.000 claims description 3
- 201000004101 esophageal cancer Diseases 0.000 claims description 3
- 208000024519 eye neoplasm Diseases 0.000 claims description 3
- 201000010175 gallbladder cancer Diseases 0.000 claims description 3
- 206010017758 gastric cancer Diseases 0.000 claims description 3
- 201000010536 head and neck cancer Diseases 0.000 claims description 3
- 208000014829 head and neck neoplasm Diseases 0.000 claims description 3
- 201000010235 heart cancer Diseases 0.000 claims description 3
- 208000024348 heart neoplasm Diseases 0.000 claims description 3
- 208000006454 hepatitis Diseases 0.000 claims description 3
- 201000010982 kidney cancer Diseases 0.000 claims description 3
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 claims description 3
- 201000008106 ocular cancer Diseases 0.000 claims description 3
- 201000002528 pancreatic cancer Diseases 0.000 claims description 3
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 3
- 201000002628 peritoneum cancer Diseases 0.000 claims description 3
- 201000002314 small intestine cancer Diseases 0.000 claims description 3
- 206010062261 spinal cord neoplasm Diseases 0.000 claims description 3
- 201000011549 stomach cancer Diseases 0.000 claims description 3
- 201000003120 testicular cancer Diseases 0.000 claims description 3
- 201000002510 thyroid cancer Diseases 0.000 claims description 3
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 3
- 206010046766 uterine cancer Diseases 0.000 claims description 3
- 241000711549 Hepacivirus C Species 0.000 claims description 2
- 230000004663 cell proliferation Effects 0.000 claims description 2
- 208000029742 colonic neoplasm Diseases 0.000 claims description 2
- 239000003112 inhibitor Substances 0.000 claims description 2
- 208000020984 malignant renal pelvis neoplasm Diseases 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 201000007444 renal pelvis carcinoma Diseases 0.000 claims description 2
- 102000007066 Prostate-Specific Antigen Human genes 0.000 claims 2
- 230000000694 effects Effects 0.000 abstract description 25
- 230000001093 anti-cancer Effects 0.000 abstract description 22
- 238000000034 method Methods 0.000 abstract description 18
- 238000011275 oncology therapy Methods 0.000 abstract 2
- 230000002411 adverse Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 106
- 241000701161 unidentified adenovirus Species 0.000 description 64
- IRSCQMHQWWYFCW-UHFFFAOYSA-N ganciclovir Chemical compound O=C1NC(N)=NC2=C1N=CN2COC(CO)CO IRSCQMHQWWYFCW-UHFFFAOYSA-N 0.000 description 38
- 229960002963 ganciclovir Drugs 0.000 description 37
- 101000655352 Homo sapiens Telomerase reverse transcriptase Proteins 0.000 description 21
- 230000001965 increasing effect Effects 0.000 description 18
- 241000699670 Mus sp. Species 0.000 description 17
- 239000007924 injection Substances 0.000 description 17
- 238000002347 injection Methods 0.000 description 17
- 210000004185 liver Anatomy 0.000 description 16
- 241000699666 Mus <mouse, genus> Species 0.000 description 12
- 241000700605 Viruses Species 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 230000005740 tumor formation Effects 0.000 description 12
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 11
- 108010082126 Alanine transaminase Proteins 0.000 description 11
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 11
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 11
- 210000003494 hepatocyte Anatomy 0.000 description 11
- GKOZUEZYRPOHIO-UHFFFAOYSA-N iridium atom Chemical compound [Ir] GKOZUEZYRPOHIO-UHFFFAOYSA-N 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 10
- 229910052741 iridium Inorganic materials 0.000 description 10
- 238000007920 subcutaneous administration Methods 0.000 description 10
- 239000013603 viral vector Substances 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 9
- 230000037396 body weight Effects 0.000 description 9
- 230000001939 inductive effect Effects 0.000 description 9
- 238000007912 intraperitoneal administration Methods 0.000 description 9
- 238000011284 combination treatment Methods 0.000 description 8
- 108010054624 red fluorescent protein Proteins 0.000 description 8
- 230000001225 therapeutic effect Effects 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 7
- 208000015181 infectious disease Diseases 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 231100000419 toxicity Toxicity 0.000 description 7
- 230000001988 toxicity Effects 0.000 description 7
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 6
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 6
- 206010067125 Liver injury Diseases 0.000 description 6
- 239000012830 cancer therapeutic Substances 0.000 description 6
- 239000005090 green fluorescent protein Substances 0.000 description 6
- 231100000234 hepatic damage Toxicity 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 6
- 230000008818 liver damage Effects 0.000 description 6
- 108091035539 telomere Proteins 0.000 description 6
- 102000055501 telomere Human genes 0.000 description 6
- 230000004614 tumor growth Effects 0.000 description 6
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 210000003411 telomere Anatomy 0.000 description 5
- 238000013518 transcription Methods 0.000 description 5
- 230000035897 transcription Effects 0.000 description 5
- 230000003612 virological effect Effects 0.000 description 5
- 108091005957 yellow fluorescent proteins Proteins 0.000 description 5
- 108091026890 Coding region Proteins 0.000 description 4
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 4
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 4
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 4
- 108091005941 EBFP Proteins 0.000 description 4
- 235000000177 Indigofera tinctoria Nutrition 0.000 description 4
- 229920000776 Poly(Adenosine diphosphate-ribose) polymerase Polymers 0.000 description 4
- 102100038358 Prostate-specific antigen Human genes 0.000 description 4
- 108091005948 blue fluorescent proteins Proteins 0.000 description 4
- -1 cationic lipid Chemical class 0.000 description 4
- 229920006317 cationic polymer Polymers 0.000 description 4
- 230000030833 cell death Effects 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 108091006047 fluorescent proteins Proteins 0.000 description 4
- 102000034287 fluorescent proteins Human genes 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 229940097275 indigo Drugs 0.000 description 4
- COHYTHOBJLSHDF-UHFFFAOYSA-N indigo powder Natural products N1C2=CC=CC=C2C(=O)C1=C1C(=O)C2=CC=CC=C2N1 COHYTHOBJLSHDF-UHFFFAOYSA-N 0.000 description 4
- 239000007928 intraperitoneal injection Substances 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 210000004940 nucleus Anatomy 0.000 description 4
- 239000002243 precursor Substances 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 108090000695 Cytokines Proteins 0.000 description 3
- 230000005778 DNA damage Effects 0.000 description 3
- 231100000277 DNA damage Toxicity 0.000 description 3
- 241000702421 Dependoparvovirus Species 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000001747 exhibiting effect Effects 0.000 description 3
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000008595 infiltration Effects 0.000 description 3
- 238000001764 infiltration Methods 0.000 description 3
- 210000003712 lysosome Anatomy 0.000 description 3
- 230000001868 lysosomic effect Effects 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 230000001172 regenerating effect Effects 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- 208000010370 Adenoviridae Infections Diseases 0.000 description 2
- 206010060931 Adenovirus infection Diseases 0.000 description 2
- 238000011729 BALB/c nude mouse Methods 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 201000009030 Carcinoma Diseases 0.000 description 2
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 2
- 206010008342 Cervix carcinoma Diseases 0.000 description 2
- 229920001661 Chitosan Polymers 0.000 description 2
- 102000000311 Cytosine Deaminase Human genes 0.000 description 2
- 108010080611 Cytosine Deaminase Proteins 0.000 description 2
- 208000009889 Herpes Simplex Diseases 0.000 description 2
- 102100037102 Homeobox protein MOX-2 Human genes 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 108010047761 Interferon-alpha Proteins 0.000 description 2
- 102000006992 Interferon-alpha Human genes 0.000 description 2
- 108060001084 Luciferase Proteins 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 108700011259 MicroRNAs Proteins 0.000 description 2
- 241000699660 Mus musculus Species 0.000 description 2
- 108700026223 Neurofibromatosis 1 Genes Proteins 0.000 description 2
- 108091000080 Phosphotransferase Proteins 0.000 description 2
- 241000714474 Rous sarcoma virus Species 0.000 description 2
- 241000713896 Spleen necrosis virus Species 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 102100033254 Tumor suppressor ARF Human genes 0.000 description 2
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 2
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- 208000011589 adenoviridae infectious disease Diseases 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 208000005266 avian sarcoma Diseases 0.000 description 2
- 238000002619 cancer immunotherapy Methods 0.000 description 2
- 230000022131 cell cycle Effects 0.000 description 2
- 201000010881 cervical cancer Diseases 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 239000000824 cytostatic agent Substances 0.000 description 2
- 230000001085 cytostatic effect Effects 0.000 description 2
- 230000002354 daily effect Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 210000001163 endosome Anatomy 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- XRECTZIEBJDKEO-UHFFFAOYSA-N flucytosine Chemical compound NC1=NC(=O)NC=C1F XRECTZIEBJDKEO-UHFFFAOYSA-N 0.000 description 2
- 229960004413 flucytosine Drugs 0.000 description 2
- 235000012631 food intake Nutrition 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000004068 intracellular signaling Effects 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 239000002679 microRNA Substances 0.000 description 2
- 230000011278 mitosis Effects 0.000 description 2
- 230000001338 necrotic effect Effects 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 238000011580 nude mouse model Methods 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 201000008968 osteosarcoma Diseases 0.000 description 2
- 108700025694 p53 Genes Proteins 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 102000020233 phosphotransferase Human genes 0.000 description 2
- 239000000651 prodrug Substances 0.000 description 2
- 229940002612 prodrug Drugs 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000002626 targeted therapy Methods 0.000 description 2
- 229940104230 thymidine Drugs 0.000 description 2
- 231100000167 toxic agent Toxicity 0.000 description 2
- 239000003440 toxic substance Substances 0.000 description 2
- 238000000844 transformation Methods 0.000 description 2
- 241001430294 unidentified retrovirus Species 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- VEEGZPWAAPPXRB-BJMVGYQFSA-N (3e)-3-(1h-imidazol-5-ylmethylidene)-1h-indol-2-one Chemical compound O=C1NC2=CC=CC=C2\C1=C/C1=CN=CN1 VEEGZPWAAPPXRB-BJMVGYQFSA-N 0.000 description 1
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- 101150084750 1 gene Proteins 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 108700001666 APC Genes Proteins 0.000 description 1
- 235000006491 Acacia senegal Nutrition 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 102100036464 Activated RNA polymerase II transcriptional coactivator p15 Human genes 0.000 description 1
- 108010056962 Adenovirus E4 Proteins Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 102400000068 Angiostatin Human genes 0.000 description 1
- 108010079709 Angiostatins Proteins 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 101150108519 CDK4 gene Proteins 0.000 description 1
- 108010076667 Caspases Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 102100031162 Collagen alpha-1(XVIII) chain Human genes 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 108010053187 Diphtheria Toxin Proteins 0.000 description 1
- 102000016607 Diphtheria Toxin Human genes 0.000 description 1
- 108010079505 Endostatins Proteins 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108090000982 GIR1 ribozyme Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 102100021519 Hemoglobin subunit beta Human genes 0.000 description 1
- 108091005904 Hemoglobin subunit beta Proteins 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 101710142888 Homeobox protein MOX-2 Proteins 0.000 description 1
- 101000746373 Homo sapiens Granulocyte-macrophage colony-stimulating factor Proteins 0.000 description 1
- 101001091385 Homo sapiens Kallikrein-6 Proteins 0.000 description 1
- 101000621309 Homo sapiens Wilms tumor protein Proteins 0.000 description 1
- 102100040018 Interferon alpha-2 Human genes 0.000 description 1
- 108010079944 Interferon-alpha2b Proteins 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 108090000174 Interleukin-10 Proteins 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 108050009288 Interleukin-19 Proteins 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 102100034866 Kallikrein-6 Human genes 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 101150115920 MTS1 gene Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000713333 Mouse mammary tumor virus Species 0.000 description 1
- 101100519207 Mus musculus Pdcd1 gene Proteins 0.000 description 1
- 208000001894 Nasopharyngeal Neoplasms Diseases 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 108700026224 Neurofibromatosis 2 Genes Proteins 0.000 description 1
- 206010061336 Pelvic neoplasm Diseases 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 108090000472 Phosphoenolpyruvate carboxykinase (ATP) Proteins 0.000 description 1
- 102100034792 Phosphoenolpyruvate carboxykinase [GTP], mitochondrial Human genes 0.000 description 1
- 101710179684 Poly [ADP-ribose] polymerase Proteins 0.000 description 1
- 102100023712 Poly [ADP-ribose] polymerase 1 Human genes 0.000 description 1
- 229920002873 Polyethylenimine Polymers 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 101000762949 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) Exotoxin A Proteins 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 102000009572 RNA Polymerase II Human genes 0.000 description 1
- 108010009460 RNA Polymerase II Proteins 0.000 description 1
- 230000004570 RNA-binding Effects 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 102100022151 Ragulator complex protein LAMTOR1 Human genes 0.000 description 1
- 108700025701 Retinoblastoma Genes Proteins 0.000 description 1
- 108010081734 Ribonucleoproteins Proteins 0.000 description 1
- 102000004389 Ribonucleoproteins Human genes 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 101150019443 SMAD4 gene Proteins 0.000 description 1
- 108091081021 Sense strand Proteins 0.000 description 1
- 241000251131 Sphyrna Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 102000003929 Transaminases Human genes 0.000 description 1
- 108090000340 Transaminases Proteins 0.000 description 1
- 108700009124 Transcription Initiation Site Proteins 0.000 description 1
- 101150046474 Vhl gene Proteins 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 239000002870 angiogenesis inducing agent Substances 0.000 description 1
- 239000004037 angiogenesis inhibitor Substances 0.000 description 1
- 229940121369 angiogenesis inhibitor Drugs 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 238000003782 apoptosis assay Methods 0.000 description 1
- FZCSTZYAHCUGEM-UHFFFAOYSA-N aspergillomarasmine B Natural products OC(=O)CNC(C(O)=O)CNC(C(O)=O)CC(O)=O FZCSTZYAHCUGEM-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 210000000013 bile duct Anatomy 0.000 description 1
- 201000009036 biliary tract cancer Diseases 0.000 description 1
- 208000020790 biliary tract neoplasm Diseases 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 235000012241 calcium silicate Nutrition 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000032677 cell aging Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 208000003167 cholangitis Diseases 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000013601 cosmid vector Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 239000002875 cyclin dependent kinase inhibitor Substances 0.000 description 1
- 229940043378 cyclin-dependent kinase inhibitor Drugs 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 231100000221 frame shift mutation induction Toxicity 0.000 description 1
- 230000037433 frameshift Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000008571 general function Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 238000010562 histological examination Methods 0.000 description 1
- 238000007489 histopathology method Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000001969 hypertrophic effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 108090000681 interleukin 20 Proteins 0.000 description 1
- 229940047122 interleukins Drugs 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 230000031146 intracellular signal transduction Effects 0.000 description 1
- 238000007917 intracranial administration Methods 0.000 description 1
- 230000002601 intratumoral effect Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 231100000832 liver cell necrosis Toxicity 0.000 description 1
- 230000007056 liver toxicity Effects 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 208000037819 metastatic cancer Diseases 0.000 description 1
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 230000000394 mitotic effect Effects 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 238000011369 optimal treatment Methods 0.000 description 1
- 108700042657 p16 Genes Proteins 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 201000010315 pericholangitis Diseases 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 208000015768 polyposis Diseases 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000000861 pro-apoptotic effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000005522 programmed cell death Effects 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000005549 size reduction Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 108010057210 telomerase RNA Proteins 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
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
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- 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
- A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
- C12N2710/10341—Use of virus, viral particle or viral elements as a vector
- C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
Definitions
- the present invention relates to a cancer-specific trans-splicing ribozyme as a gene therapy agent used in combination with radiation therapy.
- Telomerase is a ribonucleoprotein that repeatedly adds TTAGGG sequences to the ends of telomeres present at the 3' end of chromosomes to restore shortened telomere regions during DNA replication, resulting in cell perpetuity. enable proliferation. Telomerase is one of the most important enzymes that regulate the immortality and proliferative capacity of cancer cells. Hematopoietic cells and 80-90% of cancer cells have telomerase activity, while normal cells near cancer cells do not have the activity. Moreover, telomerase reactivation has a major effect on the permanent growth of advanced metastatic cancer.
- Human telomerase consists of two components: human telomerase RNA (hTR) acting as substrate RNA and human telomerase reverse transcriptase (hTERT) acting as a catalyst.
- the human telomerase reverse transcriptase (hTERT) gene is expressed in proportion to telomerase activity, and there is a strong correlation between the intracellular hTERT level and cellular telomerase activity. In particular, TERT activity is observed in more than 90% of cancer patients.
- trans-splicing ribozyme targeting hTERT attempts to develop cancer therapeutics using the trans-splicing ribozyme are being actively pursued.
- the combination of a trans-splicing ribozyme and a tissue-specific promoter exhibits high tissue specificity but very low expression efficiency, resulting in a problem in terms of therapeutic efficiency.
- telomerase activity is also shown in normal cells such as stem cells, hematopoietic stem cells, germ cells, and regenerating normal liver cells, treatment targeting hTERT is unlikely to cause toxicity to these cells.
- hepatocytes are weak, 5% of normal hepatocytes have telomerase activity, and regenerating liver is known to increase telomerase activity.
- hepatocellular carcinoma HCC is mostly accompanied by liver cirrhosis, and TERT is expressed at a low level in non-tumorous hepatocytes constituting regenerative nodules at the site of liver cirrhosis.
- Korean Patent Publication No. 10-2016-0038674 discloses a ribozyme containing a tissue-specific promoter, a trans-splicing ribozyme targeting a cancer-specific gene, and a target gene linked to the 3' exon of the ribozyme- Disclosed are a recombinant vector in which a nucleic acid sequence recognizing microRNA-122 (microRNA-122, miR-122) is additionally linked to a target gene expression cassette, and a use of a ribozyme expressed therefrom for preventing or treating liver cancer.
- the miR-122 is a microRNA known to be highly overexpressed in normal liver, and its expression decreases when liver cancer progresses.
- the prior art introduces the miR-122 target site into the 3' UTR region of the ribozyme expression vector, so that the ribozyme delivered to the liver is not expressed by the overexpressed miR-122 in the normal liver.
- the level of miR-122 is lowered, it is expressed and able to act.
- radiation therapy is widely used for various tumors as an effective method for cancer treatment, but due to problems that can cause damage to surrounding normal tissues, the upper limit of the irradiation dose is limited, and it is necessary to repeatedly irradiate several times at a low dose. Rather, there is a problem in that cancer cells acquire radiation resistance and immunosuppressive effects.
- radiation treatment for liver cancer has a problem in that radiation treatment and dose are limited due to proximity to major organs of the human body.
- the present invention is to solve the above problems, and to overcome the side effects and limitations of radiation therapy, cancer-specific trans-splicing ribozymes with excellent safety and expression efficiency as gene therapy combined with radiation therapy are provided. It has a purpose.
- cancer immunotherapy targeted therapy, and radiation therapy are used in combination.
- the present invention was completed by confirming that the side effects of each of the above treatment methods can be reduced and the cancer treatment effect can be improved when trans-splicing ribozymes targeting cancer-specific gene sequences are expressed or administered in cells did
- the present invention includes a trans-splicing ribozyme expression vector targeting a cancer-specific gene sequence, a gene delivery system comprising the vector, and/or a ribozyme expressed from the vector, and is It provides a pharmaceutical composition for preventing or treating cancer, characterized in that it is used in combination.
- the present invention provides a trans-splicing ribozyme expression vector targeting the cancer-specific gene sequence, a gene delivery system comprising the vector, and/or radiation therapy for cancer comprising the ribozyme expressed from the vector. It provides a pharmaceutical composition for enhancing the therapeutic effect.
- the present invention provides a trans-splicing ribozyme expression vector targeting a cancer-specific gene sequence, a gene delivery system comprising the vector, or a radiation-resistant cancer comprising a ribozyme expressed from the vector as an active ingredient. It provides a pharmaceutical composition for the treatment of.
- the expression vector may include a cytomegalovirus (CMV) promoter operably linked to the ribozyme gene, and splicing donor / at the 5' end of the ribozyme gene It may include a splicing donor/splicing acceptor sequence (SD/SA sequence), and may include a Woodchuck hepatitis virus Posttranscriptional Regulatory Element (WPRE) at the 3' end.
- CMV cytomegalovirus
- SD/SA sequence splicing donor/splicing acceptor sequence
- WPRE Woodchuck hepatitis virus Posttranscriptional Regulatory Element
- the expression vector may include a target gene linked to the 3' exon of the ribozyme gene.
- the expression vector contains a gene encoding a sequence complementary to part or all of microRNA-122 (microRNA-122, miR-122) at the 3'-UTR end of the ribozyme gene. can do.
- the cancer-specific gene sequence is essential for the growth, proliferation, and/or metastasis of cancer cells and is not limited as long as it is a gene that is overexpressed in cancer cells than in normal cells, but is preferably TERT (Telomerase Reverse Transcriptase) mRNA, alphafetoprotein (AFP) mRNA, carcinoembryonic antigen (CEA) mRNA, prostate-specific antigen (PSA) mRNA, cytoskeleton-associated protein 2 (CKAP2) mRNA, and mutant rat sarcoma (RAS) mRNA.
- TERT Telomerase Reverse Transcriptase
- AFP alphafetoprotein
- CEA carcinoembryonic antigen
- PSA prostate-specific antigen
- CKAP2 cytoskeleton-associated protein 2
- RAS mutant rat sarcoma
- the TERT mRNA sequence may consist of or include the nucleotide sequence of SEQ ID NO: 2.
- the trans-splicing ribozyme may consist of or include the nucleotide sequence of SEQ ID NO: 3.
- the target gene may be a cancer treatment gene or a reporter gene
- the cancer treatment gene may be a drug susceptibility gene, an apoptosis gene, a cell proliferation inhibitory gene, a cytotoxic gene, or a tumor suppressor gene. It may be at least one gene selected from the group consisting of a gene, an antigenic gene, a cytokine gene, and an angiogenesis inhibitor gene
- the drug susceptibility gene may be a Herpes Simplex Virus thymidine kinase (HSVtk) gene.
- HSVtk Herpes Simplex Virus thymidine kinase
- the HSVtk gene may consist of or include the nucleotide sequence of SEQ ID NO: 4.
- the reporter gene is luciferase, green fluorescent protein (GFP), modified green fluorescent protein (mGFP), enhanced green fluorescent protein (enhanced green fluorescent protein) EGFP), red fluorescent protein (RFP), modified red fluorescent protein (mRFP), enhanced red fluorescent protein (ERFP), blue fluorescent protein (BFP), enhanced blue fluorescent protein (EBFP), yellow fluorescent protein (YFP) ), enhanced yellow fluorescent protein (EYFP), indigo fluorescent protein (CFP) or enhanced indigo fluorescent protein (ECFP).
- GFP green fluorescent protein
- mGFP modified green fluorescent protein
- enhanced green fluorescent protein enhanced green fluorescent protein
- EGFP green fluorescent protein
- RFP red fluorescent protein
- mRFP modified red fluorescent protein
- ERFP enhanced red fluorescent protein
- BFP blue fluorescent protein
- EBFP blue fluorescent protein
- EBFP blue fluorescent protein
- EYFP enhanced yellow fluorescent protein
- CFP indigo fluorescent protein
- ECFP enhanced indigo fluorescent protein
- the gene encoding a sequence complementary to miR-122 may express a sequence complementary to part or all of miR-122, preferably a sequence complementary to all of miR-122.
- the sequence may be expressed, but is not limited to substitution, deletion or insertion of some bases as long as it binds to miR-122 in the cell and induces degradation of the ribozyme of the present invention.
- the gene encoding a sequence complementary to miR-122 may consist of or include the nucleotide sequence of SEQ ID NO: 5, and may repeatedly include the nucleotide sequence of SEQ ID NO: 5.
- the gene delivery system may be a viral vector including the expression vector, and the viral vector may be an adenoviral vector.
- the cancer to be prevented or treated according to the present invention is not limited as long as it is a disease in a state of transformation caused by abnormal proliferation of cells, and non-limiting examples thereof include liver cancer, thyroid cancer, and testicular cancer. , bone cancer, glioblastoma, ovarian cancer, brain cancer, gallbladder cancer, bile duct cancer, colorectal cancer, head and neck cancer, lymphoma, bladder cancer, leukemia, peritoneal cancer, small intestine cancer, esophageal cancer, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, eye cancer, urethral cancer, breast cancer, stomach cancer, prostate cancer, uterine cancer, lung cancer, spinal cord tumor, pancreatic cancer, and melanoma.
- the cancer may preferably be a cancer in which miR-122 is not substantially expressed in cancer tissue, and specifically, the copy number of miR-122 in cancer tissue is the copy of the ribozyme expressed in cancer tissue by the pharmaceutical composition. It can be cancer that is less than 100 times the number.
- the liver cancer is not limited to the etiology, and non-limiting examples of the etiology include hepatitis B virus, hepatitis C in which the expression of miR-122 is reduced in liver cancer tissue.
- Virus e.g., alcohol, chronic hepatitis, cirrhosis, non-alcoholic fatty liver, aflatoxin, and family history.
- the pharmaceutical composition may be administered by a route selected from the group consisting of intravenous, intraarterial, intracranial and subcutaneous, and may be administered in the form of an injection.
- the pharmaceutical composition may be administered concurrently or sequentially with radiation therapy, preferably after radiation therapy is preceded, and more preferably After performing radiation therapy 1 to 3 times, the pharmaceutical composition may be administered simultaneously with radiation therapy, sequentially, or individually several times a day.
- the present invention administers a trans-splicing ribozyme expression vector targeting a cancer-specific gene sequence in parallel with radiation therapy, a gene delivery system comprising the vector, or a ribozyme expressed from the vector to a subject It provides a cancer treatment method comprising the step of doing.
- the step in parallel with the radiation treatment may be performed simultaneously with or sequentially with the radiation treatment, but preferably, the administration step may be performed after the radiation treatment.
- the cancer treatment method of the present invention may include the following steps:
- step (2) may be performed after step (1), and step (1) may be performed after step (2). That is, the order of steps (1) and (2) above is free.
- the steps (1) to (2) may be independently repeated several times.
- step (2) is the first time at the same time when step (1) is performed once, and at the same time as the day of performing the last step (1) when step (2) is performed twice or more. can be performed
- step (1) when steps (1) and (2) are performed twice or more, step (1) may be performed every day, and step (2) may be performed every other day.
- step (2) when steps (1) and (2) are performed twice or more, respectively, step (2) may be performed for the first time on the day of performing the last step (1).
- step (1) may be to irradiate the subject with radiation of 1 to 4 Gy, preferably 1.5 to 2.5 Gy, and more preferably 2 Gy.
- the subject is not limited to any mammal in need of cancer treatment, but may be preferably a human.
- the present invention provides a trans-splicing ribozyme expression vector targeting the cancer-specific gene sequence for the production of a drug for cancer treatment used in combination with radiation therapy, a gene delivery system including the vector, or the vector Provided are the uses of ribozymes expressed from
- the trans-splicing ribozyme according to the present invention does not act on normal tissues, is specifically expressed in cancer tissues, has high safety, and has excellent expression efficiency at the post-transcriptional level, so it can be usefully used to treat cancer.
- the ribozyme according to the present invention exhibits an increased cancer treatment effect in combination with other cancer treatment methods, it can be used as a drug for the treatment of carcinoma resistant to radiation treatment, especially when used in combination with radiation treatment. As it shows a high anti-cancer effect with just a dose of radiation and a low dose of gene therapy, it can reduce side effects and increase access to treatment.
- FIG. 1 is a schematic diagram of an experiment to confirm the anti-cancer efficacy of adenovirus according to the composition of the CMV promoter-based hTERT-targeting trans-splicing ribozyme and the target gene expression cassette of the present invention and radiation combined treatment.
- FIG. 2 is a graph showing tumor size (a) and tumor weight (b) by inducing tumor formation by injecting glioblastoma cell line LN229 into nude mice and then administering ECRT-122T adenovirus.
- FIG. 3 is a graph showing tumor size by inducing tumor formation by injecting U87MG cell line, which is a glioblastoma cell, into nude mice and then administering ECRT-122T adenovirus.
- FIG. 4 is a graph showing the results of measuring AST and ALT levels after injecting ECRT-122T adenovirus into normal ICR mice.
- FIG. 5 is a graph showing the results of measuring body weight (a), food consumption (b), and liver weight (c) of mice after injecting ECRT-122T adenovirus into normal ICR mice.
- Figure 6 shows the results of histopathological examination of the liver performed after injecting different doses of ECRT-122T adenovirus into normal ICR mice.
- FIG. 7 is a graph showing the results of measuring AST and ALT levels after injecting ECRT-122T adenovirus into normal ICR mice and treating them with GCV.
- FIG. 8 is a graph showing the results of measuring body weight (a), food consumption (b), and liver weight (c) of mice after injection of ECRT-122T adenovirus into normal ICR mice and treatment with GCV.
- FIG. 9 shows the results of histopathological examination of the liver performed after injecting different doses of ECRT-122T adenovirus into normal ICR mice and treating them with GCV.
- FIG. 10 shows the results of comparing anticancer efficacy by inducing tumor formation by injecting SNU398 cells into a mouse xenograft subcutaneous model and then administering CRT-122T or ECRT-122T adenovirus.
- FIG. 11 is a schematic diagram of an experiment to confirm the anti-cancer efficacy of adenovirus according to radiation combination treatment.
- FIG. 12 shows the results of comparison of body weight changes of mice according to adenovirus administration and radiation treatment after tumor formation was induced by injecting SNU398 cells into a mouse xenograft subcutaneous model.
- FIG. 13 shows the results of comparison of changes in tumor tissue size according to adenovirus administration and radiation treatment after tumor formation was induced by injecting SNU398 cells into a mouse xenograft subcutaneous model.
- FIG. 14 shows results of comparison of tumor tissue size changes according to adenovirus administration and radiation treatment after tumor formation was induced by injecting Hep3B cells into a mouse xenograft subcutaneous model.
- 15 is a schematic diagram of an experiment for establishing an optimal adenovirus and radiation combination treatment protocol.
- Figure 16a shows the results of comparison of tumor tissue size changes after tumor formation was induced by injecting SNU398 cells into a mouse xenograft subcutaneous model, and then treated with adenovirus and/or irradiation with different usages and doses.
- 16B is a result of confirming the change in body weight of mice treated with different doses and doses of adenovirus administration and/or irradiation after tumor formation was induced by injecting SNU398 cells into a mouse xenograft subcutaneous model.
- FIG. 17 shows the results of immunofluorescence staining of rH2AX and p-ATM expression levels 4 and 48 hours after irradiation and/or RZ-001 infection in Hep3B human liver cancer cell line.
- One aspect of the present invention is
- a ribozyme-target gene expression cassette comprising a trans-splicing ribozyme targeting a cancer-specific gene sequence and a target gene linked to the 3' exon of the ribozyme;
- the expression cassette has a splicing donor/splicing donor sequence (SD/SA sequence) linked to the 5' end of the ribozyme-target gene expression cassette and WPRE linked to the 3' end,
- SD/SA sequence splicing donor/splicing donor sequence
- the recombinant vector is named ECRT-122T.
- the cytomegalovirus (CMV) promoter included in the recombinant vector according to the present invention can increase ribozyme expression in Hep3B, SNU398, and SNU449 cell lines more than the PEPCK promoter.
- CMV cytomegalovirus
- the SD/SA sequence and WPRE are included as components at both ends of the ribozyme and the target gene along with the CMV promoter, the expression efficiency of the ribozyme in vivo of the recombinant vector is higher.
- the CMV promoter, SD/SA sequence, and WPRE are simultaneously used, and a nucleic acid sequence (miR-122T) that recognizes miR-122 is additionally included to enable treatment of various cancer cells as well as liver cancer cells.
- vector is an expression vector capable of expressing a target gene in a suitable host cell, and refers to a gene construct containing essential regulatory elements operably linked to express a gene insert contained in the vector.
- operably linked refers to functional linkage between a nucleic acid expression control sequence that performs a general function and a nucleic acid sequence encoding a gene of interest.
- operably linking a ribozyme coding sequence to a promoter brings expression of the ribozyme coding sequence under the influence or control of the promoter.
- Two nucleic acid sequences (a ribozyme coding sequence and a promoter region sequence at the 5' end of the sequence) are operably linked when the action of a promoter is induced and the ribozyme coding sequence is transcribed, and the linkage between the two sequences changes the frame. If a frameshift mutation is not induced and the expression control sequence does not inhibit the expression of the ribozyme, it can be considered to be operably linked.
- Operable linkage with the recombinant vector can be prepared using genetic recombination techniques well known in the art, and site-specific DNA cutting and linking can use enzymes generally known in the art.
- the vector according to the present invention includes a signal sequence or leader sequence for membrane targeting or secretion in addition to expression control elements such as a promoter, operator, initiation codon, stop codon, polyadenylation signal, and enhancer, and can be prepared in various ways according to the purpose. there is.
- the vector's promoter may be constitutive or inducible.
- the expression vector may include a selectable marker for selecting host cells containing the vector, and may include an origin of replication in the case of a replicable expression vector. Vectors can replicate autonomously or integrate into host DNA.
- the vector according to the present invention may preferably be a plasmid vector, a cosmid vector or a viral vector, and most preferably a viral vector.
- the viral vector is preferably a retrovirus, for example, human immunodeficiency virus (HIV), mouse leukemia virus (MLV), avian sarcoma / leukemia virus (Avian sarcoma / leucosis virus (ASLV), spleen necrosis virus (SNV), Rous sarcoma virus (RSV), mouse mammary tumor virus (MMTV), adenovirus, adeno-related virus (Adeno-associated virus, AAV), or a vector derived from a herpes simplex virus (Herpes simplex virus, HSV), etc., but is not limited thereto.
- the recombinant vector according to the present invention may most preferably be a recombinant adenoviral vector.
- expression cassette used in the present invention includes a CMV promoter and a trans-splicing ribozyme-target gene, and at each end of the trans-splicing ribozyme-target gene, an SD/SA sequence and a WPRE sequence is present, and a nucleic acid sequence recognizing miR-122 is additionally linked to the 3' end of the WPRE, thereby trans-splicing ribozyme - refers to a unit cassette capable of expressing a target gene.
- the trans-splicing ribozyme-target gene expression cassette according to the present invention may further include a sequence regulating the transcription level, that is, a regulatory derivative, to the sequence linked to the trans-splicing ribozyme and the target gene.
- a sequence regulating the transcription level that is, a regulatory derivative
- SD/SA sequence splicing donor/splicing acceptor sequence
- WPRE Woodchuck hepatitis virus Posttranscriptional Regulatory Element
- a sequence recognizing miR-122 may be additionally linked to the 3' end of WPRE.
- the ribozyme-target gene expression cassette according to the present invention preferably has a splicing donor/splicing donor sequence (SD/SA) linked to the 5' end of the ribozyme, and WPRE at the 3' end of the target gene. and a sequence recognizing miR-122 may be linked to the 3' end of the WPRE.
- SD/SA splicing donor/splicing donor sequence
- SD/SA increases transcription initiation, processing of RNA polymerase II and nucleocytoplasmic export of mRNA from the nucleus to the cytoplasm, and the WPRE according to the present invention It can increase the level of pre-mRNA by increasing mRNA processing and transport from the nucleus to the cytoplasm, respectively.
- the RNA level of the ribozyme can be significantly increased in the cell to increase the death of cancer cells in vivo while allowing the specific expression of the ribozyme to be expressed in a cancer cell, thereby reducing toxicity to normal cells.
- the SD/SA sequence according to the present invention is a sequence corresponding to the beginning and end of an intron cut out in a splicing reaction to remove an intron of an RNA transcript.
- the SD sequence is a GU sequence at the 5' end of the intron
- the SA sequence may be an AG sequence at the 3' end of the intron.
- WPRE Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE)
- WPRE Posttranscriptional Regulatory Element
- the SD / SA sequence and the WPRE sequence may include the sequences of SEQ ID NO: 6 and SEQ ID NO: 7, respectively, but are not limited thereto as long as they are present in the target gene expression cassette and can promote the expression of the target gene.
- the nucleic acid sequence recognizing miR-122 is referred to as miR-122T (microRNA-122 target site) within the specification.
- the miR-122T may include the sequence of SEQ ID NO: 5 one or more times, for example, 1 to 10 times, preferably 1 to 5 times, more preferably 1 to 3 times.
- can miR-122 is normally expressed in normal hepatocytes, but its expression level is reduced in hepatoma cells. Using this, it is possible to develop a therapeutic agent with increased sensitivity and specificity for liver cancer cells.
- by linking a nucleic acid sequence recognizing miR-122 to a ribozyme to which a target gene is linked expression of a liver cancer cell-specific ribozyme can be achieved. made it possible.
- cancer-specific gene used in the present invention refers to a gene that is specifically expressed or markedly overexpressed only in cancer cells.
- the cancer-specific gene may add a feature that allows the ribozyme according to the present invention to act in a cancer-specific manner.
- cancer-specific genes are preferably TERT (Telomerase reverse transcriptase) mRNA, AFP (alphafetoprotein) mRNA, CEA (carcinoembryonic antigen) mRNA, PSA (Prostate-specific antigen) mRNA, CKAP2 (Cytoskeleton-associated protein 2) mRNA, or It may be a mutant rat sarcoma (RAS) mRNA, more preferably a telomerase reverse transcriptase (TERT) mRNA, and most preferably a human telomerase reverse transcriptase (hTERT) mRNA sequence.
- TERT Telomerase reverse transcriptase
- AFP alphafetoprotein
- CEA carcinoembryonic antigen
- PSA Prostate-specific antigen
- CKAP2 Cytoskeleton-associated protein 2
- RAS mutant rat sarcoma
- TERT telomerase reverse transcriptase
- hTERT human telomerase reverse
- the ribozyme of the present invention exhibits an anticancer effect even in cancer cell lines of tissues that do not substantially express miR-122, including the cancer-specific gene, and the present inventors are particularly interested in glioblastoma cell lines, colorectal cancer cell lines, melanoma cell lines, uterine When cervical cancer cell lines, lung cancer cell lines, osteosarcoma cell lines, breast cancer cell lines, and cholangiocarcinoma cell lines were treated with adenovirus expressing ECRT-122T, it was confirmed that cell death increased.
- TERT Telomerase reverse transcriptase
- hTERT mRNA including the sequence of SEQ ID NO: 2 may be used as a cancer-specific gene, but is not limited thereto.
- promoter is involved in the binding of RNA polymerase to initiate transcription as a portion of DNA. In general, it is located adjacent to and upstream of the target gene, and is a binding site for RNA polymerase or a transcription factor, a protein that induces RNA polymerase, and can induce the enzyme or protein to be located at the correct transcription start site. can That is, it is located at the 5' site of the gene to be transcribed in the sense strand and induces RNA polymerase to bind to the corresponding position directly or through a transcription factor to initiate mRNA synthesis for the target gene.
- a specific gene sequence have
- the promoter according to the present invention is preferably a cytomegalovirus (CMV) promoter comprising the sequence of SEQ ID NO: 1 from the viewpoint of increasing gene expression.
- CMV cytomegalovirus
- ribozyme used in the present invention is a molecule composed of an RNA molecule that acts like an enzyme or a protein containing the RNA molecule, and is also called an RNA enzyme or catalytic RNA.
- Some ribozymes inhibit their activity by cleaving their own or other RNA molecules, and other ribozymes are known to catalyze the activity of ribosome aminotransferases.
- Such ribozymes may include hammerhead ribozymes, VS ribozymes, and hairpin ribozymes.
- the ribozyme according to the present invention inhibits the activity of cancer-specific genes through the trans-splicing reaction of Group I introns, thereby exhibiting a selective anti-cancer effect, and is expressed in a conjugated form with a cancer therapeutic gene to treat cancer.
- genes can be activated. Therefore, any type of material may be used as long as it exhibits characteristics capable of inactivating cancer-specific genes and activating cancer therapeutic genes.
- the ribozyme according to the present invention may preferably be a ribozyme targeting hTERT mRNA as described above, targeting cancer cells in which hTERT is overexpressed, specifically cleaving hTERT mRNA to inhibit its expression, and generating a therapeutic gene. It can play a role in specific expression.
- trans-splicing means linking RNAs from different genes to each other.
- an hTERT target trans-splicing group I ribozyme verified for trans-splicing by recognizing cancer-specific mRNA of hTERT may be used.
- target gene used in the present invention refers to a gene whose expression is induced by being linked to mRNA of a cancer-specific gene by the ribozyme.
- the target gene according to the present invention may preferably be a gene for cancer treatment or a reporter gene, and most preferably a gene for cancer treatment.
- anti-cancer therapeutic gene refers to a polynucleotide sequence encoding a polypeptide that exhibits a therapeutic effect when expressed in cancer cells.
- the cancer therapeutic gene may be expressed in a conjugated form with the ribozyme or expressed independently to exhibit anticancer activity.
- These cancer therapeutic genes are preferably selected from the group consisting of drug susceptibility genes, apoptosis genes, cytostatic genes, cytotoxic genes, tumor suppressor genes, antigenic genes, cytokine genes, and anti-angiogenic genes. It may be one or more, and most preferably, it may be a drug susceptibility gene.
- the cancer treatment gene may be used alone or two or more genes may be used in combination.
- the drug-sensitizing gene according to the present invention is a gene encoding an enzyme that converts a non-toxic prodrug into a toxic substance, and is also called a suicide gene because cells into which the gene is introduced die. . That is, when a precursor that is not toxic to normal cells is systemically administered, the precursor is converted into a toxic metabolite only in cancer cells, thereby changing the sensitivity to the drug, thereby destroying cancer cells.
- These drug susceptibility genes are preferably HSVtk (Herpes simplex virus-thymidine kinase) genes using ganciclovir as a precursor, or E. coli using 5-fluorocytosine (5-FC) as a precursor. It may be a cytosine deaminase (CD) gene, and most preferably, it may be an HSVtk gene comprising the sequence of SEQ ID NO: 4.
- a proapoptotic gene according to the present invention refers to a nucleotide sequence that induces programmed cell death when expressed.
- Apoptosis genes known to those skilled in the art encoding p53, adenovirus E3-11.6K (derived from Ad2 and Ad5) or adenovirus E3-10.5K (derived from Ad), adenovirus E4 gene, p53 pathway gene and caspase genes may be included.
- a cytostatic gene according to the present invention refers to a nucleotide sequence that is expressed in cells and stops the cell cycle during the cell cycle. Examples include p21, retinoblastoma gene, E2F-Rb fusion protein gene, genes encoding cyclin-dependent kinase inhibitors (e.g., p16, p15, p18 and p19), growth arrest specific homeobox , GAX) gene, etc., but is not limited thereto.
- a cytotoxic gene according to the present invention refers to a nucleotide sequence that is expressed in a cell and exhibits a toxic effect. Examples include, but are not limited to, nucleotide sequences encoding Pseudomonas exotoxin, ricin toxin, diphtheria toxin, and the like.
- a tumor suppressor gene refers to a nucleotide sequence capable of suppressing a tumor phenotype or inducing apoptosis by being expressed in a target cell.
- tumor necrosis factor- ⁇ (TNF- ⁇ ), p53 gene, APC gene, DPC-4/Smad4 gene, BRCA-1 gene, BRCA-2 gene, WT-1 gene, retinoblastoma gene, MMAC- 1 gene, adenomatous polyposis coil protein, missing colon tumor (DCC) gene, MMSC-2 gene, NF-1 gene, nasopharyngeal tumor suppressor gene located on chromosome 3p21.3, MTS1 gene , CDK4 gene, NF-1 gene, NF-2 gene, VHL gene, or programmed death-1 (sPD-1).
- An antigenic gene according to the present invention refers to a nucleotide sequence that is expressed in a target cell and produces a cell surface antigenic protein that can be recognized by the immune system.
- antigenic genes known to those skilled in the art may include carcinoembryonic antigen (CEA) and p53.
- a cytokine gene according to the present invention refers to a nucleotide sequence that is expressed in a cell to produce a cytokine.
- GMCSF interleukins
- interleukins IL-1, IL-2, IL-4, IL-12, IL-10, IL-19, IL-20
- interferon ⁇ , ⁇ , ⁇ interferon ⁇ -2b
- interferon ⁇ Fusions such as -2 ⁇ -1 may be included.
- An anti-angiogenic gene according to the present invention refers to a nucleotide sequence that is expressed and releases the anti-angiogenic factor out of the cell.
- examples include angiostatin, vascular endothelial growth factor (VEGF) inhibitor, endostatin, and the like.
- HSV-tk Herpes simplex virus-thymidine kinase
- This enzyme is a representative example of a drug susceptibility gene that converts a non-toxic prodrug into a toxic substance, causing the transfected cell to die.
- the HSVtk gene is expressed in a form conjugated to the ribozyme according to the present invention and can be used as a cancer therapeutic gene exhibiting anticancer activity.
- HSVtk ⁇ ⁇ ⁇ (genbank) ⁇ AAP13943, P03176, AAA45811, P04407, Q9QNF7, KIBET3, P17402, P06478, P06479, AAB30917, P08333, BAB84107, AAP13885, AAL73990, AAG40842, BAB11942, NP_044624, NP_044492, It may be described in CAB06747 or the like.
- reporter gene is a gene used to monitor the introduction of a recombinant vector or the expression efficiency of a ribozyme according to an embodiment of the present invention, which can be monitored without damaging infected cells or tissues.
- Genes may be used without limitation.
- luciferase green fluorescent protein (GFP), modified green fluorescent protein (mGFP), enhanced green fluorescent protein (EGFP), red fluorescent protein (RFP), Modified red fluorescent protein (mRFP), enhanced red fluorescent protein (ERFP), blue fluorescent protein (BFP), enhanced blue fluorescent protein (EBFP), yellow fluorescent protein (YFP), enhanced yellow fluorescence protein (EYFP), indigo fluorescent protein (CFP) or enhanced indigo fluorescent protein (ECFP).
- the expression level of the cancer cell-specific ribozyme can be observed.
- the ribozyme of the present invention contains a promoter and a microRNA target site, so it is not expressed in normal cells and is specific to cancer cells. can manifest. It is obvious to those skilled in the art that it can be applied to diagnose whether cancer has occurred in a specific tissue using this.
- Another aspect of the present invention is a gene delivery system comprising a recombinant vector according to the present invention.
- the term “gene delivery system” refers to a system capable of increasing expression efficiency by increasing the transfer efficiency of a desired gene and/or nucleic acid sequence into a cell, and is a virus-mediated system. and non-viral systems.
- Virus-mediated systems use viral vectors such as retroviral vectors and adenovirus vectors, and have relatively higher efficiency of intracellular gene transfer than non-viral systems because they use the unique intracellular penetration mechanism of viruses that infect human cells. It is known.
- the non-viral vector after entering the cell, the non-viral vector has a problem in that the endosome fuses with the lysosome and then the genes are degraded in the endolysosome, but the viral vector does not pass through the lysosome and transfers the gene into the nucleus by a mechanism It has the advantage of high gene delivery efficiency because the loss of is small.
- Viral vectors that can be used in the present invention may be vectors derived from retroviruses, adenoviruses, adeno-associated viruses, and the like, as described above for recombinant vectors. These viral vectors can be assembled into viral particles and then introduced into cells by a transduction method such as infection.
- a recombinant adenovirus containing the above-described recombinant vector as an example of a gene transfer vehicle was constructed. That is, the recombinant adenovirus performs a function of delivering a recombinant vector expressing a trans-splicing ribozyme specific to a cancer-specific gene to a target cell (eg, cancer cell), and the recombinant delivered into the cell Vectors are expressed by the intracellular transcription system.
- the expressed trans-splicing ribozyme can insert a target gene linked to the ribozyme into the transcript of a cancer-specific gene present in large numbers in cancer cells.
- an adenovirus containing a recombinant vector (ECRT-122T) expressing the trans-splicing ribozyme of the present invention is named RZ-001.
- the non-viral system is a method using a cationic lipid delivery system or a cationic polymer carrier or the like as a delivery medium for nucleic acids and/or genes, or using an electroporation method.
- Cationic lipid carriers use the positive charge of nanometer-sized liposomes or lipid nanoparticles mainly composed of cationic lipids to form complexes with negatively charged genes, expression vectors or nucleic acids containing genes, and then form the complexes by phagocytosis. method of delivery into cells. Complexes delivered into cells are first delivered from endosomes to lysosomes and then released into the cytosol to be expressed.
- Cationic polymer carriers deliver genes in a similar way to cationic lipid carriers, except that polymers are used instead of lipids.
- Representative cationic polymers include polyethyleneimine, poly-L-lysine, and chitosan. (chitosan), etc.
- a complex formed by combining the recombinant vector of the present invention with a cationic lipid carrier or a cationic polymer carrier can be used as a gene carrier.
- the gene delivery system includes the recombinant vector described above, and both virus-mediated systems and non-viral systems may be used, but it is preferable to use a virus-mediated system.
- Another aspect of the present invention is a ribozyme expressed from a recombinant vector according to the present invention.
- Another aspect of the present invention is a recombinant vector according to the present invention, a gene delivery system comprising the recombinant vector, or a pharmaceutical composition for preventing or treating cancer comprising a ribozyme as an active ingredient.
- the pharmaceutical composition may be used in combination with radiation therapy, or may be used to enhance the efficacy of radiation therapy.
- cancer used in the present invention refers to a problem in the control function of normal division, differentiation, and death of cells, which abnormally proliferates and infiltrates surrounding tissues and organs to form lumps and destroy existing structures. denotes a deformed state.
- Non-limiting examples of cancer according to the present invention include liver cancer, thyroid cancer, testicular cancer, bone cancer, glioblastoma, ovarian cancer, brain cancer, gallbladder cancer, biliary tract cancer, colorectal cancer, head and neck cancer, lymphoma, bladder cancer, leukemia, peritoneal cancer, small intestine cancer. , esophageal cancer, renal pelvic cancer, kidney cancer, heart cancer, duodenal cancer, eye cancer, urethral cancer, breast cancer, stomach cancer, prostate cancer, uterine cancer, lung cancer, spinal cord tumor, pancreatic cancer, and melanoma, more preferably liver cancer, glioblastoma , and bile duct cancer, etc., preferably liver cancer.
- a cancer according to the present invention may be a cancer resistant to radiation therapy.
- the cancer according to the present invention may preferably have a copy number (expression level) of miR-122 expressed in cancer tissue that is less than 100 times the copy number of the ribozyme expressed in cancer tissue by the pharmaceutical composition. .
- the ratio of miR-122 to ribozyme will increase.
- the amount of adenovirus expressing the ribozyme can be determined by inferring the amount of ribozyme to exhibit the anticancer effect according to the expression level of miR-122 in the cancer tissue.
- the function (expression) of the ribozyme having a miR-122 target site is weakened. It was confirmed that high anticancer efficacy can be obtained when the copy number of the ribozyme expressed in cancer tissue by the pharmaceutical composition according to is less than 100 times.
- the cancer according to the present invention may preferably be a cancer in which miR-122 is not substantially expressed in cancer tissues.
- the above “cancer in which miR-122 is not substantially expressed in cancer tissue” means that although miR-122 is expressed in cancer tissue, it is expressed in cancer tissue to the extent that it does not substantially affect the function of the ribozyme having a miR-122 target site. It means cancer with a low copy number of miR-122.
- the ribozyme according to the present invention is used in colorectal cancer, glioblastoma, melanoma, cervical cancer, lung cancer, osteosarcoma, breast cancer and cholangiocarcinoma cell lines in which miR-122 is not substantially expressed in cancer tissues. Efficacy was confirmed.
- liver cancer according to the present invention is preferably hepatitis B virus, hepatitis C virus in which the expression of miR-122 is reduced in liver cancer tissues, alcohol, chronic hepatitis, cirrhosis, nonalcoholic fatty liver disease, aflatoxin It may be due to one or more causes selected from the group consisting of, and family history.
- the present invention can provide the above-described recombinant vector, gene delivery system, and ribozyme as a pharmaceutical composition used in combination with radiation therapy.
- the pharmaceutical composition of the present invention can be used in combination with radiation therapy to effectively induce tumor tissue size reduction even with a small dose of the pharmaceutical composition and a low radiation dose.
- by administering the pharmaceutical composition of the present invention after radiation treatment 1 Secondarily, by reducing the size of tumor tissues by radiation to increase tissue penetration of drugs, and by inducing death of cancer cells resistant to radiation by the pharmaceutical composition of the present invention, effective cancer treatment is possible.
- the pharmaceutical composition of the present invention may be administered simultaneously or sequentially with radiation therapy as a combination with radiation therapy, but preferably, the pharmaceutical composition of the present invention may be administered after radiation therapy, If radiation therapy is preceded, subsequent treatment may be performed at the same time as administration of the pharmaceutical composition of the present invention and radiation therapy.
- the pharmaceutical composition of the present invention can be provided for the treatment of radiation-resistant cancer.
- 'simultaneously' means that the radiation treatment and administration of the pharmaceutical composition of the present invention are performed within 24 hours, and 'sequentially' means that they are performed over 24 hours.
- prevention used in the present invention refers to any activity that inhibits the proliferation, metastasis, or acquisition of aggressiveness of cancer cells or delays the onset of cancer by administering the pharmaceutical composition according to the present invention.
- treatment refers to any activity in which cancer is improved or its symptoms are beneficially changed by administration of a composition containing a vector according to the present invention.
- the pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent.
- pharmaceutically acceptable carriers, excipients and diluents that can be used in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, and alginate.
- gelatin calcium phosphate, calcium silicate, calcium carbonate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like.
- the pharmaceutical composition of the present invention may be administered orally or parenterally depending on the desired method, but is preferably administered parenterally.
- the pharmaceutical composition according to the present invention can be directly administered intravenously, intraarterially, into cancer tissue or subcutaneously, or administered as an injection.
- the injection according to the present invention may be in a form dispersed in a sterile medium so that it can be used as it is when administered to a patient, or may be administered after dispersing in an appropriate concentration by adding distilled water for injection.
- it when prepared as an injection, it may be mixed with buffers, preservatives, analgesics, solubilizers, tonicity agents, stabilizers, etc., and may be prepared in unit dosage ampoules or multiple dosage forms.
- the dosage of the pharmaceutical composition of the present invention varies depending on the condition and body weight of the patient, the severity of the disease, the drug type, the administration route and time, but can be appropriately selected by those skilled in the art. Meanwhile, the pharmaceutical composition according to the present invention may be used alone or in combination with auxiliary treatment methods such as surgical treatment.
- a conventional trans-splicing ribonucleic acid targeting the +21 site of hTERT mRNA, having an antisense sequence (SEQ ID NO: 8) of 326 nucleotides in length, and having HSV-tk as a therapeutic gene A CMV promoter was introduced into the zyme. Transcription efficiency was increased by inserting an SV40 intron splicing donor/acceptor (SD/SA) sequence between the CMV promoter and the ribozyme.
- SD/SA SV40 intron splicing donor/acceptor
- WPRE a posttranscriptional regulatory element of Woodchuck Hepatitis Virus, was inserted into the 3' region of HSV-tk, a therapeutic gene, to increase protein expression efficiency of the therapeutic gene, and to the 3' end of the construct. 3 copies of miR-122T were inserted so that it could be regulated by miR-122.
- Tumor formation was induced by subcutaneous injection of 1x10 7 (100 ⁇ l) of LN229 cell line or 5x10 6 (100 ⁇ l) of U87MG cell line into 6-week-old male Balb/c-nunu mice.
- adenovirus expressing ECRT-122T was injected at a dose of 1.0x10 9 VP (100 ⁇ l) once every 2 days for a total of 3 times.
- GCV was injected intraperitoneally (Intraperitoneal injection, IP injection) (20 times in total) at a dose of 50 mg/kg twice a day for 10 days from 24 hours after the first virus injection.
- ECRT-122T ribozyme expressed from ECRT-122T can be effectively applied to various carcinomas that do not express miR-122 in addition to liver cancer.
- ECRT-122T when ECRT-122T is administered systemically or locally as an anticancer agent for other cancers other than liver cancer, it can be introduced into the normal liver. In this case, the action of miR-122T will suppress the induction of toxicity in the normal liver.
- ECRT-122T adenovirus was injected into normal ICR mice once, and AST and ALT levels were measured on days 15 and 29 after injection.
- mice Normal ICR mice were injected with ECRT-122T adenovirus, and GCV was intraperitoneally administered twice a day for 10 days, and then AST and ALT levels were measured.
- AST and ALT levels were intravenously administered at a dose of 2.5x10 10 VP/head, and GCV was administered twice a day for 10 days, hepatocyte necrosis and inflammation were induced, and liver damage was induced. appeared to be However, when 0.25x10 10 VP/head and 1.0 x 10 10 VP/head were administered alone or in combination with GCV, AST and ALT levels related to liver damage increased up to 15 days after administration, but histological examination performed on day 29 showed No significant toxicological changes considered to be related to adenovirus administration were observed in .
- Comparative Example 1 Comparison of anti-cancer efficacy of CRT-122T and ECRT-122T
- Tumor formation is induced by injecting SNU398 cells into a mouse xenograft subcutaneous model, and when the tumor grows beyond a certain size, adenovirus containing the CRT-122T or ECRT-122T vector is administered at a dose of 1x10 9 VP.
- adenovirus containing the CRT-122T or ECRT-122T vector is administered at a dose of 1x10 9 VP.
- a total of 2 times was injected into the cancer tissue (intratumoral injection, IT injection), and GCV was administered intraperitoneally at a dose of 50 mg/kg twice a day for 10 days from 24 hours after the first adenovirus injection (Intraperitoneal injection). , IP injection) (total 20 times).
- the tumor size and body weight were measured every 3 days while the mice were bred, and after 22 days, the mice were sacrificed and the final tumor size, liver weight, AST (aspartate transaminase) and ALT (alanine transaminase) levels were measured.
- FIGS. 10a and 10b it was confirmed that the anticancer efficacy of ECRT-122T was superior to that of CRT-122T. There was no significant difference in the body weight and liver weight of mice between the experimental groups, and it was found from the AST and ALT levels that adenovirus did not induce liver toxicity (FIGS. 10c to 10e).
- SNU398 (Fig. 13) cells and Hep3B (Fig. 14) were applied to a mouse xenograft subcutaneous model (6-7 week old male BALB/C nude mouse). Tumor formation was induced by injecting the cells, and when the tumors reached 100 mm 3 , 2 gray of iridium (IR) was injected every day for 3 days, followed by adenovirus administration. On the last day of radiation treatment, after the last radiation, an adenovirus vector was administered once a day, 3 times at 2-day intervals (Adenovirus dose: 1x10 9 VP per dose).
- GCV ganciclovir
- Tumor formation was induced by injecting SNU398 cells into a mouse xenograft subcutaneous model (6-7 week old male BALB/C nude mouse), and when tumors reached 100 mm 3 , adenovirus administration and irradiation were performed under various conditions. conducted. Radiation was administered at 2 Gy once, and adenovirus was administered at 1x10 9 VP/head. Irradiation and adenovirus administration of each group were conducted under the following conditions:
- G3 RZ-001 alone. A total of 3 doses at 2-day intervals
- G5 radiation alone. A total of 3 irradiation at 2Gy 1 day intervals
- G6 1 dose of RZ-001 on the same day after irradiation of 2Gy once
- G7 A total of 3 doses of RZ-001 every 2 days from the day after irradiation of 2Gy once
- G8 Radiation 2Gy 1 day intervals, a total of 3 times irradiation. One dose of RZ-001 on the same day after the last irradiation
- G9 Radiation 2Gy 1 day interval, a total of 3 times irradiation. From the day after the last irradiation, RZ-001 was administered a total of 3 times at 2-day intervals.
- GCV ganciclovir
- ganciclovir was intraperitoneally administered at 50 mg/kg daily for 10 days 24 hours after the last radiation treatment in the radiation-only treatment group and 24 hours after the first adenovirus treatment in the adenovirus-only or combined treatment group (FIG. 14).
- Adenovirus administration was RZ-001, and the experiment was divided into a radiation and adenovirus combined treatment group (IR+RZ-001) and a radiation or adenovirus alone treatment group, and the efficacy was compared by measuring the mouse weight and tumor size. did
- the radiation and adenovirus combination treatment group significantly reduced tumor growth compared to the untreated, radiation or adenovirus treatment group alone.
- Intracellular signaling changes after irradiation and/or RZ-001 infection in Hep3B human liver cancer cell line were investigated.
- the adenovirus-only treatment group infected Hep3B cells with 40 moi of adenovirus without GCV treatment, and the irradiation combination treatment group was irradiated with 4 Gy of radiation and then infected with 40 moi of adenovirus.
- Changes in intracellular signaling over time without GCV treatment confirmed. That is, intracellular changes by the single action of the hTERT-targeted ribozyme and the combined use with radiation were confirmed under radiation alone and GCV untreated conditions.
- RZ-001 expressing ECRT-122T and Ad-Mock not expressing ribozyme were used as a control group (FIG. 16(A)).
- the rH2AX expression level was confirmed by staining with an anti-rH2AX antibody.
- an increase in rH2AX spot was confirmed 48 hours after RZ-001 infection, indicating that genomic DNA instability increased due to the decrease in TERT expression by RZ-001 infection.
- an increase in rH2AX spot was confirmed 4 hours after irradiation in all samples, including radiation-only cells, indicating that genomic DAN instability increased by irradiation.
- genomic DAN repair proceeded through reduction of rH2AX spot in cells treated with radiation alone, but rH2AX spot was maintained in cells treated with radiation + RZ-001 infection. It can be seen that the genomic DNA instability effect continues (FIG. 16(B)). On the other hand, since the increase and maintenance of rH2AX spot was not observed in the radiation + Ad-mock treatment group, it can be seen that the result is a specific phenomenon caused by ribozyme expression rather than a non-specific reaction caused by adenovirus infection.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Veterinary Medicine (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
Histopathology /Groups |
G1 | G2 | G3 | G4 | G5 | G6 |
Vehicle (PBS) |
PBS+GCV | 1.0x1010
VP
+GCV |
2.5x1010
VP
+GCV |
0.25x1010
VP
+GCV |
1.0x1010 VP | |
No. examined | 5 | 5 | 5 | 5 | 5 | 5 |
No specific lesion | 5 (100) | 4 (80.0) | 3 (60.0) | 0 (0.00) | 5 (100) | 3 (60.0) |
Cell infiltration, mononuclear or mixed cells, multifocal | 0 (0.00) | 1 (20.0) | 1 (20.0) | 4 (80.0) | 0 (0.00) | 0 (0.00) |
Grades: Minimal | 0 | 1 | 1 | 0 | 0 | 0 |
Mild | 0 | 0 | 0 | 4 | 0 | 0 |
Microabcess, focal | 0 (0.00) | 0 (0.00) | 1 (20.0) | 0 (0.00) | 0 (0.00) | 0 (0.00) |
Grades: Minimal | 0 | 0 | 1 | 0 | 0 | 0 |
Hepatocytomegaly, diffuse | 0 (0.00) | 0 (0.00) | 0 (0.00) | 5 (100) | 0 (0.00) | 0 (0.00) |
Grades: Minimal | 0 | 0 | 0 | 1 | 0 | 0 |
Mild | 0 | 0 | 0 | 2 | 0 | 0 |
Moderate | 0 | 0 | 0 | 2 | 0 | 0 |
Necrotic hepatocytes | 0 (0.00) | 0 (0.00) | 0 (0.00) | 3 (60.0) | 0 (0.00) | 0 (0.00) |
Grades: Minimal | 0 | 0 | 0 | 2 | 0 | 0 |
Mild | 0 | 0 | 0 | 1 | 0 | 0 |
Hepatocytic mitosis, increased, diffuse | 0 (0.00) | 0 (0.00) | 0 (0.00) | 4 (80.0) | 0 (0.00) | 1 (20.0) |
Grades: Minimal | 0 | 0 | 0 | 2 | 0 | 1 |
Mild | 0 | 0 | 0 | 1 | 0 | 0 |
Moderate | 0 | 0 | 0 | 1 | 0 | 0 |
Oval cell hyperplasia | 0 (0.00) | 0 (0.00) | 0 (0.00) | 3 (60.0) | 0 (0.00) | 0 (0.00) |
Grades: Minimal | 0 | 0 | 0 | 3 | 0 | 0 |
Pericholangitis, (multi)focal | 0 (0.00) | 0 (0.00) | 0 (0.00) | 3 (60.0) | 0 (0.00) | 1 (20.0) |
Grades: Minimal | 0 | 0 | 0 | 3 | 0 | 1 |
Group | Treatment | Route of administration | Dose | Number of Dose | Dose volume |
G1 | PBS | i.t | - | 3 | 30 mL |
GCV | i.p | 50mg/kg | 10 | 100 mL | |
G2 | RZ-001 | i.t | 1 x 109 VP | 1 | 30 mL |
GCV | i.p | 50mg/ kg | 10 | 100 mL | |
G3 | RZ-001 | i.t | 1 x 109 VP | 3 | 30 mL |
GCV | i.p | 50mg/ kg | 10 | 100mL | |
G4 | IR | - | 2 Gy | 1 | - |
PBS | i.t | - | 30 mL | ||
GCV | i.p | 50mg/ kg | 10 | 100mL | |
G5 | IR | - | 2 Gy | 3 | - |
PBS | i.t | - | 30 mL | ||
GCV | i.p | 50mg/ kg | 10 | 100mL | |
G6 | IR | 2 Gy | 1 | - | |
RZ-001 | i.t | 1 x 109 VP | 1 | 30 mL | |
GCV | i.p | 50mg/ kg | 10 | 100 mL | |
G7 | IR | 2 Gy | 1 | - | |
RZ-001 | i.t | 1 x 109 VP | 3 | 30 mL | |
GCV | i.p | 50mg/ kg | 10 | 100mL | |
G8 | IR | 2 Gy | 3 | - | |
RZ-001 | i.t | 1 x 109 VP | 1 | 30 mL | |
GCV | i.p | 50mg/ kg | 10 | 100 mL | |
G9 | IR | 2 Gy | 3 | - | |
RZ-001 | i.t | 1 x 109 VP | 3 | 30 mL | |
GCV | i.p | 50mg/ kg | 10 | 100mL |
SEQ No. | Feature | Sequence (5' → 3') |
1 | CMV promoter sequence | gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgc |
2 | hTERT mRNA sequence | caggcagcgc tgcgtcctgc tgcgcacgtg ggaagccctg gccccggcca cccccgcgat gccgcgcgct ccccgctgcc gagccgtgcg ctccctgctg cgcagccact accgcgaggt gctgccgctg gccacgttcg tgcggcgcct ggggccccag ggctggcggc tggtgcagcg cggggacccg gcggctttcc gcgcgctggt ggcccagtgc ctggtgtgcg tgccctggga cgcacggccg ccccccgccg ccccctcctt ccgccaggtg tcctgcctga aggagctggt ggcccgagtg ctgcagaggc tgtgcgagcg cggcgcgaag aacgtgctgg ccttcggctt cgcgctgctg gacggggccc gcgggggccc ccccgaggcc ttcaccacca gcgtgcgcag ctacctgccc aacacggtga ccgacgcact gcgggggagc ggggcgtggg ggctgctgct gcgccgcgtg ggcgacgacg tgctggttca cctgctggca cgctgcgcgc tctttgtgct ggtggctccc agctgcgcct accaggtgtg cgggccgccg ctgtaccagc tcggcgctgc cactcaggcc cggcccccgc cacacgctag tggaccccga aggcgtctgg gatgcgaacg ggcctggaac catagcgtca gggaggccgg ggtccccctg ggcctgccag ccccgggtgc gaggaggcgc gggggcagtg ccagccgaag tctgccgttg cccaagaggc ccaggcgtgg cgctgcccct gagccggagc ggacgcccgt tgggcagggg tcctgggccc acccgggcag gacgcgtgga ccgagtgacc gtggtttctg tgtggtgtca cctgccagac ccgccgaaga agccacctct ttggagggtg cgctctctgg cacgcgccac tcccacccat ccgtgggccg ccagcaccac gcgggccccc catccacatc gcggccacca cgtccctggg acacgccttg tcccccggtg tacgccgaga ccaagcactt cctctactcc tcaggcgaca aggagcagct gcggccctcc ttcctactca gctctctgag gcccagcctg actggcgctc ggaggctcgt ggagaccatc tttctgggtt ccaggccctg gatgccaggg actccccgca ggttgccccg cctgccccag cgctactggc aaatgcggcc cctgtttctg gagctgcttg ggaaccacgc gcagtgcccc tacggggtgc tcctcaagac gcactgcccg ctgcgagctg cggtcacccc agcagccggt gtctgtgccc gggagaagcc ccagggctct gtggcggccc ccgaggagga ggacacagac ccccgtcgcc tggtgcagct gctccgccag cacagcagcc cctggcaggt gtacggcttc gtgcgggcct gcctgcgccg gctggtgccc ccaggcctct ggggctccag gcacaacgaa cgccgcttcc tcaggaacac caagaagttc atctccctgg ggaagcatgc caagctctcg ctgcaggagc tgacgtggaa gatgagcgtg cgggactgcg cttggctgcg caggagccca ggggttggct gtgttccggc cgcagagcac cgtctgcgtg aggagatcct ggccaagttc ctgcactggc tgatgagtgt gtacgtcgtc gagctgctca ggtctttctt ttatgtcacg gagaccacgt ttcaaaagaa caggctcttt ttctaccgga agagtgtctg gagcaagttg caaagcattg gaatcagaca gcacttgaag agggtgcagc tgcgggagct gtcggaagca gaggtcaggc agcatcggga agccaggccc gccctgctga cgtccagact ccgcttcatc cccaagcctg acgggctgcg gccgattgtg aacatggact acgtcgtggg agccagaacg ttccgcagag aaaagagggc cgagcgtctc acctcgaggg tgaaggcact gttcagcgtg ctcaactacg agcgggcgcg gcgccccggc ctcctgggcg cctctgtgct gggcctggac gatatccaca gggcctggcg caccttcgtg ctgcgtgtgc gggcccagga cccgccgcct gagctgtact ttgtcaaggt ggatgtgacg ggcgcgtacg acaccatccc ccaggacagg ctcacggagg tcatcgccag catcatcaaa ccccagaaca cgtactgcgt gcgtcggtat gccgtggtcc agaaggccgc ccatgggcac gtccgcaagg ccttcaagag ccacgtctct accttgacag acctccagcc gtacatgcga cagttcgtgg ctcacctgca ggagaccagc ccgctgaggg atgccgtcgt catcgagcag agctcctccc tgaatgaggc cagcagtggc ctcttcgacg tcttcctacg cttcatgtgc caccacgccg tgcgcatcag gggcaagtcc tacgtccagt gccaggggat cccgcagggc tccatcctct ccacgctgct ctgcagcctg tgctacggcg acatggagaa caagctgttt gcggggattc ggcgggacgg gctgctcctg cgtttggtgg atgatttctt gttggtgaca cctcacctca cccacgcgaa aaccttcctc aggaccctgg tccgaggtgt ccctgagtat ggctgcgtgg tgaacttgcg gaagacagtg gtgaacttcc ctgtagaaga cgaggccctg ggtggcacgg cttttgttca gatgccggcc cacggcctat tcccctggtg cggcctgctg ctggataccc ggaccctgga ggtgcagagc gactactcca gctatgcccg gacctccatc agagccagtc tcaccttcaa ccgcggcttc aaggctggga ggaacatgcg tcgcaaactc tttggggtct tgcggctgaa gtgtcacagc ctgtttctgg atttgcaggt gaacagcctc cagacggtgt gcaccaacat ctacaagatc ctcctgctgc aggcgtacag gtttcacgca tgtgtgctgc agctcccatt tcatcagcaa gtttggaaga accccacatt tttcctgcgc gtcatctctg acacggcctc cctctgctac tccatcctga aagccaagaa cgcagggatg tcgctggggg ccaagggcgc cgccggccct ctgccctccg aggccgtgca gtggctgtgc caccaagcat tcctgctcaa gctgactcga caccgtgtca cctacgtgcc actcctgggg tcactcagga cagcccagac gcagctgagt cggaagctcc cggggacgac gctgactgcc ctggaggccg cagccaaccc ggcactgccc tcagacttca agaccatcct ggactgatgg ccacccgccc acagccaggc cgagagcaga caccagcagc cctgtcacgc cgggctctac gtcccaggga gggaggggcg gcccacaccc aggcccgcac cgctgggagt ctgaggcctg agtgagtgtt tggccgaggc ctgcatgtcc ggctgaaggc tgagtgtccg gctgaggcct gagcgagtgt ccagccaagg gctgagtgtc cagcacacct gccgtcttca cttccccaca ggctggcgct cggctccacc ccagggccag cttttcctca ccaggagccc ggcttccact ccccacatag gaatagtcca tccccagatt cgccattgtt cacccctcgc cctgccctcc tttgccttcc acccccacca tccaggtgga gaccctgaga aggaccctgg gagctctggg aatttggagt gaccaaaggt gtgccctgta cacaggcgag gaccctgcac ctggatgggg gtccctgtgg gtcaaattgg ggggaggtgc tgtgggagta aaatactgaa tatatgagtt tttcagtttt gaaaaaaa |
3 | hTERT targeting trans-splicing ribozyme | ggcaggaaaa gttatcaggc atgcacctgg tagctagtct ttaaaccaat agattgcatc ggtttaaaag gcaagaccgt caaattgcgg gaaaggggtc aacagccgtt cagtaccaag tctcagggga aactttgaga tggccttgca aagggtatgg taataagctg acggacatgg tcctaaccac gcagccaagt cctaagtcaa cagatcttct gttgatatgg atgcagttca cagactaaat gtcggtcggg gaagatgtat tcttctcata agatatagtc ggacctctcc ttaatgggag ctagcggatg aagtgatgca acactggagc cgctgggaac taatttgtat gcgaaagtat attgattagt tttggagtac tcg |
4 | HSV-tk | atggcttcgt acccctgcca tcaacacgcg tctgcgttcg accaggctgc gcgttctcgc ggccatagca accgacgtac ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc cgcctggagc agaaaatgcc cacgctactg cgggtttata tagacggtcc tcacgggatg gggaaaacca ccaccacgca actgctggtg gccctgggtt cgcgcgacga tatcgtctac gtacccgagc cgatgactta ctggcaggtg ctgggggctt ccgagacaat cgcgaacatc tacaccacac aacaccgcct cgaccagggt gagatatcgg ccggggacgc ggcggtggta atgacaagcg cccagataac aatgggcatg ccttatgccg tgaccgacgc cgttctggct cctcatatcg ggggggaggc tgggagctca catgccccgc ccccggccct caccctcatc ttcgaccgcc atcccatcgc cgccctcctg tgctacccgg ccgcgcgata ccttatgggc agcatgaccc cccaggccgt gctggcgttc gtggccctca tcccgccgac cttgcccggc acaaacatcg tgttgggggc ccttccggag gacagacaca tcgaccgcct ggccaaacgc cagcgccccg gcgagcggct tgacctggct atgctggccg cgattcgccg cgtttacggg ctgcttgcca atacggtgcg gtatctgcag ggcggcgggt cgtggcggga ggattgggga cagctttcgg ggacggccgt gccgccccag ggtgccgagc cccagagcaa cgcgggccca cgaccccata tcggggacac gttatttacc ctgtttcggg cccccgagtt gctggccccc aacggcgacc tgtacaacgt gtttgcctgg gccttggacg tcttggccaa acgcctccgt cccatgcacg tctttatcct ggattacgac caatcgcccg ccggctgccg ggacgccctg ctgcaactta cctccgggat ggtccagacc cacgtcacca cccccggctc cataccgacg atctgcgacc tggcgcgcac gtttgcccgg gagatggggg aggctaactg a |
5 | miR-122T | caaacaccat tgtcacactc ca |
6 | SD/SA sequence | agatctgaac tgaaaaacca gaaagttaac tggtaagttt agtctttttg tcttttattt caggtcccgg atccggtggt ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt tacttctagg cctgtacgga agtgttactt ctgctctaaa agctgcggaa ttgtacccag g |
7 | WPRE sequence | aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgc |
8 | ribozyme antisense sequence | aaggccagca cgttcttcgc gccgcgctcg cacagcctct gcagcactcg ggccaccagc tccttcaggc aggacacctg gcggaaggag ggggcggcgg ggggcggccg tgcgtcccag ggcacgcaca ccaggcactg ggccaccagc gcgcggaaag ccgccgggtc cccgcgctgc accagccgcc agccctgggg ccccaggcgc cgcacgaacg tggccagcgg cagcacctcg cggtagtggc tgcgcagcag ggagcgcacg gctcggcagc ggggagcgcg cggcatcgcg ggggtggccg gggccagggc ttccca |
9 | Beta-globin poly(A) signal | aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctca |
10 | ribozyme forward primer | ttccggagga cagacacatc ga |
11 | ribozyme reverse primer | gcagataccg caccgtattg gc |
Claims (18)
- 암 특이적 유전자 서열을 표적으로 하는 트랜스-스플라이싱 리보자임 발현 벡터, 상기 벡터를 포함하는 유전자 전달 시스템, 또는 상기 벡터로부터 발현된 리보자임을 포함하며,방사선 치료와 병용되는 것을 특징으로 하는, 암의 예방 또는 치료용 약학적 조성물.
- 제1항에 있어서,상기 발현 벡터는 상기 리보자임 유전자와 작동가능하게 연결된 사이토메갈로바이러스 (cytomegalovirus, CMV) 프로모터를 포함하고,상기 리보자임 유전자 5' 말단 위치에 스플라이싱 공여/스플라이싱 수여 서열(splicing donor/splicing acceptor sequence, SD/SA sequence)를 포함하며,3' 말단 위치에 WPRE(Woodchuck hepatitis virus Posttranscriptional Regulatory Element)를 포함하는 것을 특징으로 하는, 약학적 조성물.
- 제1항에 있어서,상기 암 특이적 유전자 서열은 TERT(Telomerase Reverse Transcriptase) mRNA, AFP(alphafetoprotein) mRNA, CEA(carcinoembryonic antigen) mRNA, PSA(Prostate-specific antigen) mRNA, CKAP2(Cytoskeleton-associated protein 2) mRNA 및 돌연변이 RAS(Rat sarcoma) mRNA로 이루어진 군에서 선택되는 것인, 약학적 조성물.
- 제1항에 있어서,상기 트랜스-스플라이싱 리보자임은 서열번호 3의 핵산 서열을 포함하는 것인, 약학적 조성물.
- 제1항에 있어서,상기 발현 벡터는 상기 리보자임 유전자의 3' 엑손에 연결된 목적 유전자를 포함하는 것을 특징으로 하는, 약학적 조성물.
- 제5항에 있어서,상기 목적 유전자는 암 치료용 유전자 또는 리포터 유전자인 것을 특징으로 하는, 약학적 조성물.
- 제6항에 있어서,상기 암 치료용 유전자는 약제 감수성 유전자, 세포사멸 유전자, 세포증식 억제 유전자, 세포독성 유전자, 종양 억제인자 유전자, 항원성 유전자, 사이토카인 유전자 및 항신생 혈관 생성 유전자로 이루어진 군에서 선택되는 것인, 약학적 조성물.
- 제7항에 있어서,상기 약제 감수성 유전자는 HSVtk(Herpes Simplex Virus thymidine kinase) 유전자인 것을 특징으로 하는, 약학적 조성물.
- 제1항에 있어서,상기 발현 벡터는 리보자임 유전자 3'-UTR 말단 위치에 마이크로 RNA-122(microRNA-122, miR-122)의 일부 또는 전부와 상보적인 서열을 암호화하는 유전자를 포함하는 것을 특징으로 하는, 약학적 조성물.
- 제9항에 있어서,상기 miR-122와 상보적인 서열을 암호화하는 유전자는 서열번호 5의 염기서열을 1 카피 이상 포함하는 것을 특징으로 하는, 약학적 조성물.
- 제1항에 있어서,상기 암은 간암, 갑상선암, 고환암, 골암, 교모세포종, 난소암, 뇌암, 담낭암, 담도암, 대장암, 두경부암, 림프종, 방광암, 백혈병, 복막암, 소장암, 식도암, 신우암, 신장암, 심장암, 십이지장암, 안구암, 요도암, 유방암, 위암, 전립선암, 자궁암, 폐암, 척수종양, 췌장암, 및 흑색종으로 이루어진 군으로부터 선택되는 1종 이상의 암인 것을 특징으로 하는, 약학적 조성물.
- 제1항에 있어서,상기 암은 암 조직에서 miR-122가 실질적으로 발현되지 않는 암인 것을 특징으로 하는, 약학적 조성물.
- 제12항에 있어서,상기 암은 간암이고,상기 간암은 B형 간염 바이러스(Hepatitis B virus), 간암 조직에서 miR-122의 발현이 저하되는 C형 간염 바이러스, 알코올, 만성 간염, 간경변, 비알코올성 지방간, 아플라톡신, 및 가족력으로 이루어진 군으로부터 선택되는 어느 하나 이상을 원인으로 하는 것을 특징으로 하는, 약학적 조성물.
- 제1항에 있어서,상기 약학적 조성물은 방사선 치료와 동시에 또는 순차로 투여되는 것을 특징으로 하는, 약학적 조성물.
- 제14항에 있어서,상기 약학적 조성물은 방사선 치료 1 내지 3 회 수행 이후에 순차로 투여되는 것을 특징으로 하는, 약학적 조성물.
- 제15항에 있어서,상기 약학적 조성물은 1일 1회, 1 내지 3회 반복 투여되는 것을 특징으로 하는, 약학적 조성물.
- 제16항에 있어서,상기 약학적 조성물을 2회 이상 투여하는 경우 격일로 투여되는 것을 특징으로 하는, 약학적 조성물.
- 암 특이적 유전자 서열을 표적으로 하는 트랜스-스플라이싱 리보자임 발현 벡터, 상기 벡터를 포함하는 유전자 전달 시스템, 또는 상기 벡터로부터 발현된 리보자임을 유효성분으로 포함하는 방사선 내성 암의 치료용 약학적 조성물.
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202280067233.6A CN118139649A (zh) | 2021-08-06 | 2022-08-08 | 作为与放射线治疗联合使用的基因治疗剂的癌症特异性反式剪接核酶 |
EP22853581.1A EP4382135A1 (en) | 2021-08-06 | 2022-08-08 | Cancer-specific trans-splicing ribozyme as gene therapy product combined with radiotherapy |
JP2024507127A JP2024530190A (ja) | 2021-08-06 | 2022-08-08 | 放射線治療と併用される遺伝子治療剤として癌特異的なトランススプライシングリボザイム |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR20210103814 | 2021-08-06 | ||
KR10-2021-0103814 | 2021-08-06 | ||
KR1020220098089A KR20230022814A (ko) | 2021-08-06 | 2022-08-05 | 방사선 치료와 병용되는 유전자 치료제로서 암 특이적 트랜스-스플라이싱 리보자임 |
KR10-2022-0098089 | 2022-08-05 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2023014206A1 true WO2023014206A1 (ko) | 2023-02-09 |
Family
ID=85154660
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2022/011774 WO2023014206A1 (ko) | 2021-08-06 | 2022-08-08 | 방사선 치료와 병용되는 유전자 치료제로서 암 특이적 트랜스-스플라이싱 리보자임 |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP4382135A1 (ko) |
JP (1) | JP2024530190A (ko) |
WO (1) | WO2023014206A1 (ko) |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015194853A1 (ko) * | 2014-06-18 | 2015-12-23 | 한국원자력연구원 | 모링가잎 추출물을 포함하는 방사선 병용 처리용 조성물 및 이를 이용한 치료방법 |
KR20160038674A (ko) | 2014-09-29 | 2016-04-07 | 단국대학교 산학협력단 | 암 특이적 트랜스-스플라이싱 라이보자임 및 이의 용도 |
KR20190007488A (ko) * | 2016-05-13 | 2019-01-22 | 리제너론 파아마슈티컬스, 인크. | 암을 치료하기 위한 항-pd1 항체 및 방사선의 조합 |
KR20200102943A (ko) * | 2019-02-22 | 2020-09-01 | 알지노믹스 주식회사 | 암 특이적 트랜스-스플라이싱 리보자임 및 이의 용도 |
KR102195221B1 (ko) * | 2019-12-31 | 2020-12-24 | 서울대학교산학협력단 | 포스파티딜이노시톨 3-키나아제 억제제 및 프로그램화 세포 사멸 단백질 1 억제제를 포함하는, 삼중음성 유방암의 방사선 병용 치료용 약학적 조성물 |
-
2022
- 2022-08-08 EP EP22853581.1A patent/EP4382135A1/en active Pending
- 2022-08-08 WO PCT/KR2022/011774 patent/WO2023014206A1/ko active Application Filing
- 2022-08-08 JP JP2024507127A patent/JP2024530190A/ja active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015194853A1 (ko) * | 2014-06-18 | 2015-12-23 | 한국원자력연구원 | 모링가잎 추출물을 포함하는 방사선 병용 처리용 조성물 및 이를 이용한 치료방법 |
KR20160038674A (ko) | 2014-09-29 | 2016-04-07 | 단국대학교 산학협력단 | 암 특이적 트랜스-스플라이싱 라이보자임 및 이의 용도 |
KR20190007488A (ko) * | 2016-05-13 | 2019-01-22 | 리제너론 파아마슈티컬스, 인크. | 암을 치료하기 위한 항-pd1 항체 및 방사선의 조합 |
KR20200102943A (ko) * | 2019-02-22 | 2020-09-01 | 알지노믹스 주식회사 | 암 특이적 트랜스-스플라이싱 리보자임 및 이의 용도 |
KR102195221B1 (ko) * | 2019-12-31 | 2020-12-24 | 서울대학교산학협력단 | 포스파티딜이노시톨 3-키나아제 억제제 및 프로그램화 세포 사멸 단백질 1 억제제를 포함하는, 삼중음성 유방암의 방사선 병용 치료용 약학적 조성물 |
Non-Patent Citations (1)
Title |
---|
"Final report", 26 January 2018, DANKOOK UNIVERSITY, Korea, article LEE SEONGWOOK: "Development of self-regulation targeting gene therapy technology after gene transcription and transcription", pages: 1 - 154, XP093032293 * |
Also Published As
Publication number | Publication date |
---|---|
EP4382135A1 (en) | 2024-06-12 |
JP2024530190A (ja) | 2024-08-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR102252423B1 (ko) | 암 특이적 트랜스-스플라이싱 리보자임 및 이의 용도 | |
KR101712247B1 (ko) | 암 특이적 트랜스-스플라이싱 라이보자임 및 이의 용도 | |
US20010053768A1 (en) | Gene therapy using replication competent targeted adenoviral vectors | |
Van Putten et al. | Sitimagene ceradenovec: a gene-based drug for the treatment of operable high-grade glioma | |
Weihl et al. | Gene therapy for cerebrovascular disease | |
WO2018106068A1 (ko) | 세포외 기질 분해 인자를 발현하는 재조합 아데노바이러스를 포함하는 항암용 조성물 | |
US11078487B2 (en) | Cancer-specific trans-splicing ribozymes and use thereof | |
Bhattacharyya et al. | Gene therapy developments for pancreatic cancer | |
US7491525B2 (en) | Specific proliferation in tumour cell which can express antioncogene with high efficiency and the use of it | |
WO2023014206A1 (ko) | 방사선 치료와 병용되는 유전자 치료제로서 암 특이적 트랜스-스플라이싱 리보자임 | |
Shen et al. | Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene | |
WO2023128048A1 (ko) | 암 특이적 트랜스-스플라이싱 리보자임과 면역관문억제제를 포함하는 암 치료적 병용물 | |
Rosenfeld et al. | Gene therapy strategies for novel cancer therapeutics | |
KR20230022814A (ko) | 방사선 치료와 병용되는 유전자 치료제로서 암 특이적 트랜스-스플라이싱 리보자임 | |
KR20230030063A (ko) | 암 특이적 트랜스-스플라이싱 리보자임과 키나아제 억제제를 포함하는 암 치료적 병용물 | |
WO2016052851A1 (ko) | 암 특이적 트랜스-스플라이싱 라이보자임 및 이의 용도 | |
WO2020013617A1 (ko) | 항종양 조성물 | |
CN114144231A (zh) | 用于治疗癌症的crispr方法 | |
WO2022158891A1 (ko) | 면역관문 억제제를 발현하는 암 특이적 트랜스-스플라이싱 리보자임 및 이의 용도 | |
KR100357054B1 (ko) | 종양 특이증식 재조합 아데노바이러스 및 그를 이용하여 종양세포만을 특이적으로 형질전환시키는 방법 | |
WO2022060155A1 (ko) | 재조합 발생이 최소화된 유전자치료 벡터, 상기 벡터를 포함하는 재조합 레트로바이러스 및 상기 재조합 레트로바이러스를 포함하는 암의 예방 또는 치료용 약학 조성물 | |
US8945531B2 (en) | Recombinant herpes virus and pharmaceutical composition containing the same | |
CN118139649A (zh) | 作为与放射线治疗联合使用的基因治疗剂的癌症特异性反式剪接核酶 | |
Nagayama | Gene therapy for thyroid cancer | |
Zhang et al. | Section Review—Oncologic, Endocrine & Metabolic: Gene Therapy Strategies for Cancer |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 22853581 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2024507127 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2022853581 Country of ref document: EP Effective date: 20240306 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 202280067233.6 Country of ref document: CN |