AU2004212856B2 - Anticancer agent comprising LK8 protein as an active ingredient - Google Patents
Anticancer agent comprising LK8 protein as an active ingredient Download PDFInfo
- Publication number
- AU2004212856B2 AU2004212856B2 AU2004212856A AU2004212856A AU2004212856B2 AU 2004212856 B2 AU2004212856 B2 AU 2004212856B2 AU 2004212856 A AU2004212856 A AU 2004212856A AU 2004212856 A AU2004212856 A AU 2004212856A AU 2004212856 B2 AU2004212856 B2 AU 2004212856B2
- Authority
- AU
- Australia
- Prior art keywords
- protein
- cancer
- metastasis
- tumor
- cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
- 108090000623 proteins and genes Proteins 0.000 title claims description 88
- 102000004169 proteins and genes Human genes 0.000 title claims description 81
- 239000002246 antineoplastic agent Substances 0.000 title description 22
- 239000004480 active ingredient Substances 0.000 title description 7
- 206010028980 Neoplasm Diseases 0.000 claims description 42
- 206010027476 Metastases Diseases 0.000 claims description 27
- 230000009401 metastasis Effects 0.000 claims description 27
- 201000011510 cancer Diseases 0.000 claims description 16
- 206010009944 Colon cancer Diseases 0.000 claims description 13
- 210000004185 liver Anatomy 0.000 claims description 12
- 206010060862 Prostate cancer Diseases 0.000 claims description 10
- 201000001441 melanoma Diseases 0.000 claims description 10
- 206010038038 rectal cancer Diseases 0.000 claims description 10
- 210000004072 lung Anatomy 0.000 claims description 8
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 7
- 208000015634 Rectal Neoplasms Diseases 0.000 claims description 7
- 201000001275 rectum cancer Diseases 0.000 claims description 7
- 208000029742 colonic neoplasm Diseases 0.000 claims description 6
- 210000001072 colon Anatomy 0.000 claims description 5
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- 201000005202 lung cancer Diseases 0.000 claims description 4
- 208000020816 lung neoplasm Diseases 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 2
- 239000004615 ingredient Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 235000018102 proteins Nutrition 0.000 description 75
- 210000004027 cell Anatomy 0.000 description 32
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 241000699666 Mus <mouse, genus> Species 0.000 description 15
- 230000000694 effects Effects 0.000 description 12
- 238000002512 chemotherapy Methods 0.000 description 11
- 102100040214 Apolipoprotein(a) Human genes 0.000 description 9
- 241000700159 Rattus Species 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 101710115418 Apolipoprotein(a) Proteins 0.000 description 7
- 230000014509 gene expression Effects 0.000 description 7
- 102100036826 Aldehyde oxidase Human genes 0.000 description 6
- 101000928314 Homo sapiens Aldehyde oxidase Proteins 0.000 description 6
- 239000013604 expression vector Substances 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000000546 pharmaceutical excipient Substances 0.000 description 6
- 238000001959 radiotherapy Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 230000004614 tumor growth Effects 0.000 description 5
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000034994 death Effects 0.000 description 4
- 231100000517 death Toxicity 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 3
- 241000235058 Komagataella pastoris Species 0.000 description 3
- 241001529936 Murinae Species 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- 239000002257 antimetastatic agent Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 231100000371 dose-limiting toxicity Toxicity 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 201000005296 lung carcinoma Diseases 0.000 description 3
- 230000003387 muscular Effects 0.000 description 3
- 238000011580 nude mouse model Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 208000020615 rectal carcinoma Diseases 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 108010012927 Apoprotein(a) Proteins 0.000 description 2
- 238000011740 C57BL/6 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
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 101000889990 Homo sapiens Apolipoprotein(a) Proteins 0.000 description 2
- 108010007622 LDL Lipoproteins Proteins 0.000 description 2
- 102000007330 LDL Lipoproteins Human genes 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 230000033115 angiogenesis Effects 0.000 description 2
- 238000011888 autopsy Methods 0.000 description 2
- 238000010876 biochemical test Methods 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 235000014121 butter Nutrition 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 238000001415 gene therapy Methods 0.000 description 2
- 230000002489 hematologic effect Effects 0.000 description 2
- 238000001727 in vivo Methods 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
- 239000013028 medium composition Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- -1 olive oil Chemical class 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 230000008488 polyadenylation Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000012453 sprague-dawley rat model Methods 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 229910021654 trace metal Inorganic materials 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- 108700028369 Alleles 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
- 102000006991 Apolipoprotein B-100 Human genes 0.000 description 1
- 108010008150 Apolipoprotein B-100 Proteins 0.000 description 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 1
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 1
- 239000004606 Fillers/Extenders Substances 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
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 1
- 102100021866 Hepatocyte growth factor Human genes 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- 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
- 108010033266 Lipoprotein(a) Proteins 0.000 description 1
- 206010028116 Mucosal inflammation Diseases 0.000 description 1
- 201000010927 Mucositis Diseases 0.000 description 1
- 238000011794 NU/NU nude mouse Methods 0.000 description 1
- 208000003788 Neoplasm Micrometastasis Diseases 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102100027378 Prothrombin Human genes 0.000 description 1
- 108010094028 Prothrombin Proteins 0.000 description 1
- 239000012979 RPMI medium Substances 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 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
- 208000024313 Testicular Neoplasms Diseases 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 244000299461 Theobroma cacao Species 0.000 description 1
- 235000005764 Theobroma cacao ssp. cacao Nutrition 0.000 description 1
- 235000005767 Theobroma cacao ssp. sphaerocarpum Nutrition 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 1
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 1
- 230000007059 acute toxicity Effects 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- 238000011047 acute toxicity test Methods 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000003527 anti-angiogenesis Effects 0.000 description 1
- 230000001399 anti-metabolic effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 238000009534 blood test Methods 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000000038 chest Anatomy 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 238000011254 conventional chemotherapy Methods 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- 238000010410 dusting Methods 0.000 description 1
- 230000000487 effect on differentiation Effects 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000010005 growth-factor like effect Effects 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- VMPHSYLJUKZBJJ-UHFFFAOYSA-N lauric acid triglyceride Natural products CCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCC)COC(=O)CCCCCCCCCCC VMPHSYLJUKZBJJ-UHFFFAOYSA-N 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 230000001926 lymphatic effect Effects 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 208000037819 metastatic cancer Diseases 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 201000003152 motion sickness Diseases 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 108010085336 phosphoribosyl-AMP cyclohydrolase Proteins 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002250 progressing effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 201000001514 prostate carcinoma Diseases 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 229940039716 prothrombin Drugs 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000011780 sodium chloride 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
- 239000005720 sucrose Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 238000011521 systemic chemotherapy Methods 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 229960005356 urokinase Drugs 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- 239000007222 ypd medium Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Gastroenterology & Hepatology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Marine Sciences & Fisheries (AREA)
- Oncology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Description
WO 2004/073730 PCT/KR2004/000357 ANTICANCER AGENT COMPRISING LK8 PROTEIN AS AN ACTIVE INGREDIENT FIELD OF THE INVENTION The present invention relates to an anticancer agent comprising a certain protein as an active ingredient, more precisely, to an anticancer agent comprising a protein corresponding to kringle KV38 of apolipoprotein(a) as an active ingredient.
BACKGROUND
A tumor is developed by uncontrollable disordered abnormal cell proliferation. If this tumor shows a destructive growth, invasiveness and metastasis, it is regarded as a malignant tumor.
Invasiveness is a character to infiltrate or destroy surrounding tissues, and in particular, a basal layer forming a boundary of tissues is destroyed by the character, resulting in the local spread and sometimes inflow of a tumor through circulatory system. Metastasis means the spread of tumor cells from the original birthplace to other areas through lymphatic or blood vessels.
1 WO 2004/073730 PCT/KR2004/000357 In a broad sense, metastasis also means the direct extension of tumor cells through peritoneal cavity or other space.
As of today, surgical operation, radiotherapy and chemotherapy have been used for the treatment of a cancer singly or jointly. The surgical operation is a way to remove diseased tissues.
Thus, tumors in specific regions such as breast, colon and skin can be effectively removed by the surgical operation. However, a tumor in vertebra or dispersive tumor like leukemia cannot be properly treated by the surgical operation.
Chemotherapy blocks cell replication or metabolism, and has been used for the treatment of breast cancer, lung cancer and testis cancer.
Though, patients with cancers who have been treated by chemotherapy have seriously suffered from the side effects of systemic chemotherapy.
Motion sickness and vomiting, are common but serious examples of all. The side effects of chemotherapy can even affect the life of a patient since they might drop the adaptability of a patient rapidly. Besides, Dose Limiting Toxicity (DLT) is also one of major side effects of WO 2004/073730 PCT/KR2004/000357 chemotherapy, which draws a careful attention in the administration of a medicine. Mucositis is an example of DLT against anticancer agents such as which is an antimetabolic cytotoxic agent, and methotrexate, and anticancer antibiotics like doxorubicin. If a patient suffers seriously from such side effects of chemotherapy, he or she should be hospitalized and given an anodyne for reducing pain. So, side effects of chemotherapy and radiotherapy are the biggest problem for the treatment of cancer patients.
Therefore, it is an urgent need to develop an anticancer agent originated from a living creature in order to reduce side effects of chemotherapy.
In particular, among substances generated in a living creature, a promising subject has been found which does not attack cancer cells directly but prevents cancer cells from growing by working toward various endothelial cells helping cancer cell growth. So, an anticancer agent containing the subject can not only treat cancers but also prevent metastasis.
Kringle is a kind of a protein structure WO 2004/073730 PCT/KR2004/000357 which is composed of 80 amino acids and three intramolecular disulfide bonds. Kringle structure is found in many proteins such as prothrombin (Walz, D.A. et al., Proc. Natl. Acad. Sci., 74:1069-1073, 1977), urokinase (Pennica, D. et al., Nature, 301:579-582, 1983), hepatocyte growth factor (Lukker, N. A. et al., Protein Eng., 7:895- 903, 1994) and apolipoprotein(a) (refered as 'apo(a)' hereinafter) (McLean, J.W. et al., Nature, 330:132-137, 1987). Kringle is composed of an individual folding unit. The functions of kringle have not been clearly explained, yet.
Apo(a) includes two types of kringle domains, KIV and KV, and an inactive protease-like region.
The kringle domain KIV is divided into 10 subtypes (KIV-1 KIV-10) according to the homology of amino acids, and 15 40 copy numbers of the domain are found in various human alleles of the apo(a) gene. Apo(a) forms a lipoprotein(a) (referred as hereinafter) by covalent bond with apo B-100, a major protein component of lowdensity lipoprotein (LDL) (Fless, J. Biol.
Chem., 261:8712-8717, 1986). The increase of Lp(a) content in cytoplasm itself is a major risk factor of artherosclerosis (Armstrong, V.W. et al., WO 2004/073730 PCT/KR2004/000357 Artherosclerosis, 62:249-257, 1986; Assmann, G., Am. J. Cardiol., 77:1179-1184, 1996).
Based on the fact that artherosclerosis and cancer cell growth depend on angiogenesis, the present inventors have studied on an anticancer activity of KV38, one of human apo(a) kringles.
As a result, the present inventors have completed this invention by confirming that the protein can be effectively used as an anticancer agent because it inhibits angiogenesis by an endogenous growth factor like bFGF which is necessary for cancer cell growth.
SUMMARY OF THE INVENTION It is an object of this invention to provide an anticancer agent comprising a human apo(a) kringle KV38 (referred as 'LK8 protein' hereinafter) as an active ingredient.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS In order to achieve the above object of the present invention, the present invention provides WO 2004/073730 PCT/KR2004/000357 an anticancer agent comprising a LK8 protein as an active ingredient.
Hereinafter, the present invention is described in detail.
The anticancer agent of the present invention is characterized by including LK8 protein having an amino acid sequence represented by SEQ. ID. No 1 as an active ingredient. It is preferred to use the agent as a metastasis inhibitor, more preferably, it is used for the suppression of metastasis of colon carcinoma or rectal cancer to liver.
In addition, the anticancer agent is preferably used for the treatment of primary tumors. More preferably, the agent is used for the treatment of a cancer selected from a group consisting of prostate cancer, lung cancer, colon cancer and rectal cancer.
It is preferred for the anticancer agent of the present invention to contain LK8 protein by 0.1 100 mg/kg, more preferred to contain LK8 protein by 1 50 mg/kg, and administration times are 1 4 per day. But the composition is not WO 2004/073730 PCT/KR2004/000357 limited thereto, and is possibly changed according to the conditions of a patient, and types and progressing speed of a disease.
In this invention, "KV38" means an apo(a) kringle, and "LK8" means a recombinant protein of KV38. However, both KV38 and LK8 protein are called as LK8 protein in general unless mentioned specifically.
LK8 protein of the present invention is a domain corresponding to KV38 kringle of many apo(a) kringle domains and has an inhibiting effect on proliferation and differentiation of cancer cells and metastasis by suppressing the activity of endothelial cells in vitro and in vivo as well. As explained in the preferred embodiment of the present invention, the systemic administration of LK8 protein results in the inhibition of a primary tumor and its metastasis (see FIG. 2 FIG. Therefore, LK8 protein of the present invention can be effectively used as an anticancer agent especially for primary tumors and as a metastasis inhibitor owing to its functions of suppressing tumor growth and metastasis.
WO 2004/073730 PCT/KR2004/000357 The treatment effect will be enhanced if LK8 protein of the present invention is used together with conventional chemotherapy or radiotherapy.
Radiotherapy is performed to destroy a primary tumor, and if LK8 protein is administered during radiotherapy, metastasis can be prevented more effectively. As for chemotherapy, cytotoxicity caused by huge dosage of chemical anticancer agents is the biggest problem. If LK8 protein of the invention is administered during chemotherapy, the decreased amount of chemical anticancer agents will bring equivalent or even improved anticancer effects as well as lessen the cytotoxicity.
In conclusion, if the administration of LK8 protein is performed together with surgical operation, radiotherapy, chemotherapy or immunotherapy, the treatment effect will be maximized. And further, the continuous administration of LK8 protein extends dormancy of micrometastasis, suppresses the growth of a primary tumor and stabilizes conditions. Many conventional anticancer agents are designed for long term administration, causing troubles such as continuous production of a protein and high price WO 2004/073730 PCT/KR2004/000357 of the product. The alternative to them is gene therapy. And LK8 protein is also expected to maximize the effect of an anticancer agent or a metastasis inhibitor if it is used in gene therapy.
The anticancer agent comprising LK8 protein of the present invention can be administered orally or parenterally and be used in general forms of pharmaceutical formulation.
The anticancer agent can be prepared for oral or parenteral administration by mixing with generally used fillers, extenders, binders, wetting agents, disintegrating agents, diluents such as surfactants, or excipients. Solid formulations for oral administration are tablets, pills, dusting powders, granules and capsules.
These solid formulations are prepared by mixing one or more suitable excipients such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. Except for the simple excipients, lubricants, for example magnesium stearate, talc, etc, can be used. Liquid formulations for oral administrations are suspensions, solutions, emulsions and syrups, and the above mentioned formulations can contain various excipients such WO 2004/073730 PCT/KR2004/000357 as wetting agents, sweeteners, aromatics and preservatives in addition to generally used simple diluents such as water and liquid paraffin.
Formulations for parenteral administration are sterilized aqueous solutions, water-insoluble excipients, suspensions, emulsions, and suppositories. Water insoluble excipients and suspensions can contain, in addition to the active compound or compounds, propylene glycol, polyethylene glycol, vegetable oil like olive oil, injectable ester like ethylolate, etc.
Suppositories can contain, in addition to the active compound or compounds, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerinated gelatin, etc.
LD
50 of the LK8 protein is about 1,000 ig/kg, suggesting that the anticancer agent of the present invention is very much safe (see Table 2).
BRIEF DESCRIPTION OF THE DRAWINGS The application of the preferred embodiments of the present invention is best understood with reference to the accompanying drawings, wherein: WO 2004/073730 PCT/KR2004/000357 FIG. 1 is a schematic representation of the expression vector 'pMBRI-LK8 (8.25 kb)' of LK8 gene, in which LK8 cDNA (261 bp) is inserted between AOX1 promoter and AOX1 terminator, FIG. 2 is a set of photographs and a graph showing that the pulmonary metastasis of a murine melanoma cell line, B16F10, which was injected into mouse (C57BL/6) tail vein, is inhibited by the treatment of LK8 protein, Lung of the mouse treated with PBS only, Lung of the mouse treated with LK8 protein by 1 mg/kg, A graph showing that metastasis of B16F10 cells to lung of the mouse is inhibited by the treatment of LK8 protein, FIG. 3 is a set of photographs and a graph showing that the metastasis of liver by a mouse colorectal cancer cell line CT-26 transplanted into mouse spleen is inhibited by the treatment of LK8 protein, Liver of the mouse treated with PBS (control) and LK8 protein (10 mg/kg/day), A graph showing the colony number of CT- WO 2004/073730 PCT/KR2004/000357 26 cells metastasized into liver of the mouse treated with PBS (control) and LK8 protein (10 mg/ kg/day), photographs showing the distribution of CT-26 cancer cells metastasized into liver of the mouse treated with PBS (control) and LK8 protein mg/kg/day), which is observed by hematoxylin eosin staining, FIG. 4 is a set of graphs showing the changes of the size of a tumor in a mouse transplanted with human prostate carcinoma PC-3 cells, according to the administration of LK8 protein, A graph showing the changes of the size of a tumor after treating LK8 protein by 100 mg/kg /day, A graph showing the changes of the size of a tumor after treating LK8 protein by 50 mg/kg /day, FIG. 5 is a graph showing the changes of the size of a tumor in a mouse transplanted with human lung carcinoma A549 cells, according to the administration of LK8 protein, WO 2004/073730 PCT/KR2004/000357 FIG. 6 is a set of graphs showing the changes of the size of a tumor and the weight of a tumor in mice transplanted with human rectal and colon carcinoma LS 174T cells, according to the administration of LK8 protein.
EXAMPLES
Practical and presently preferred embodiments of the present invention are illustrative as shown in the following Examples.
However, it will be appreciated that those skilled in the art, on consideration of this disclosure, may make modifications and improvements within the spirit and scope of the present invention.
Example 1: Preparation of LK8 protein Construction of LK8 expression vector pMBRI- LK8 In order to prepare LK8 protein effectively, the present inventors constructed LK8 expression vector first.
WO 2004/073730 PCT/KR2004/000357 For the expression of LK8 gene, pPIC9 vector kb, Invitrogen, Netherland) was used as a basic vector. As seen in the representative map, pPIC9 expression vector includes promoter AOX1 which provides high expression by methanol, afactor secretion signal which enables the secretion of an expressed protein, 3'AOX1(TT), AOX1 polyadenylation signal, which enables effective termination of transcription and polyadenylation, and a DNA fragment coding histidinol dehydrogenase of wild type Pichia pastoris used as a selectable marker for a transformant of the above strain, in order.
At first, LK8 gene was amplified by PCR with primers 'LK8N-Xhol' represented by SEQ. ID. No 2 and 'LK8C-EcoRI' represented by SEQ. ID. No 3, using pET15b/LK8 (see PCT/KR99/00554) as a template. The PCR product was digested with restriction enzymes Xhol and EcoRI, followed by subcloning by inserting the product into pPIC9 vector which was digested with the same restriction enzymes. Finally, the LK8 gene expression vector 'pMBRI-LK8 (8.25 kb) was constructed (FIG. 1).
WO 2004/073730 PCT/KR2004/000357 Preparation of a transformant including pMBRI-LK8 Pichia pastoris, a methylotrophic yeast, was used as a host for the preparation of a recombinant transformant.
Particularly, LK8 gene expression vector 'pMBRI-LK8' was treated with a restriction enzyme SacI, leading to linearization. The vector was inserted into AOXI gene of the above host strain chromosome by homologous recombination. At that time, electroporation was performed for transformation. A recombinant yeast transformant was selected from histidine-deficient medium by examining colony forming. PCR was performed to confirm if LK8 cDNA was inserted into AOX1 region of chromosome of the selected recombinant transformant. And then, the recombinant transformant was cultured and the expression of LK8 gene was induced by methanol. As a result, the expression of LK8 gene was confirmed, suggesting that the protein was mass-secreted in the culture medium.
The secreted LK8 protein of the present invention was composed of amino acid sequence WO 2004/073730 PCT/KR2004/000357 represented by SEQ. ID. No 1.
Cultivation of a recombinant strain Seed culture In this invention, a recombinant strain was obtained by inserting LK8 gene into Pichia pastoris. The established strain was seedcultured for 24 hours to obtain appropriate biomass and activity (when it was diluted by times, ODeo6 was 0.8 1.2).
Seed culture was performed in YDP medium (1% yeast extract, 2% peptone, 2% dextrose) for 24 hours by shaking culture. 75 L fermenter was used.
The volume of a beginning medium was 20 L and the final volume of culture solution was adjusted to L by fed-batch culture method.
Main culture After completing seed culture in YPD medium, main culture was performed with seed culture solution as much as 30% out of the primary medium.
Main culture was performed in a fermenter containing ingredients seen in the below table 1.
When the ferment was saturated by the supply of 16 WO 2004/073730 PCT/KR2004/000357 methanol, the ferment was recovered more than for the production of LK8 protein. In the meantime, methanol was being supplied continuously to induce the continuous expression of the protein.
The procedure was repeated to produce LK8 protein.
Consumption speed of a carbon source was in proportion to the amount of cells. So, when some of ferment was recovered, the speed of methanol supply was automatically regulated not to be out of From the repetition of the above culture process, during which fermentation was continued for over 200 hours, the secreted LK8 protein was obtained by 250 mg per 1 L of culture solution.
<Table 1> Medium composition for main culture Kinds of medium Component Concentration Medium for main Methanol 500 g/L culture Trace metal 8 mW/L solution WO 2004/073730 PCT/KR2004/000357 Medium composition CuSO4 5H 2 0 4 g/L of trace metal KI 0.3 g/L solution MnS04 H 2 0 4 g/L 0.1 g/L NaMo •H 2 0 0.1 g/L H3BO 4 0.01 g/L
H
3 B0 4 oCl 2 0.1 g/L 3 g/L ZnC1 2 0 g/L FeSO 4 HzO 0.1 g/L Biotin 5 m1/L 5 m-K/L
H
2 S0 4 Example 2: Experimental lung metastasis by intravenous administration of B16F10 murine melanoma cells B16F10 cells (1.8 X 105), mouse melanoma (referred as 'melanoma cells' hereinafter) (American Type Culture Collection), were administered in tail vein of a C57BL/6 mouse (Charles River Japan, Inc.). From the next day, LK8 protein, prepared in the above <Example 1>, was injected subcutaneously twice (1 mg/kg/day, 0.2 mg/kg/day) a day for 14 days. A control group was injected with PBS instead of LK8 protein. On the 1 3 t h day from cell transplantation, the mouse was dissected, and lung was taken to count the number of colony of the metastasized cancer cells (melanoma cells).
WO 2004/073730 PCT/KR2004/000357 As a result, huge colonies were formed in lungs of the control group mice injected with PBS, suggesting the transfer of melanoma cells (FIG.
2a). On the contrary, the number and the size of colonies generated by the transfer of melanoma cells were much less and smaller in the experimental group mice injected with LK8 protein (FIG. 2b). In conclusion, the group treated with LK8 protein by 1 mg/kg showed 53% metastasis inhibition, comparing to the control group (FIG.
2c).
Example 3: Experimental liver metastasis by intrasplenic administration of murine colorectal cancer cell CT-26 CT-26 cells (American Type Culture Collection), mouse colorectal cancer cells, were injected in spleen to induce metastasis into liver.
Then, metastasis-inhibiting effect of LK8 protein was investigated. Particularly, CT-26 cells, which were 80% grown up on a plate, were washed with PBS, followed by singularizing with 0.02% WO 2004/073730 PCT/KR2004/000357 EDTA. The single cells were washed with PBS again, then resuspended in PBS carefully. The suspension was stained with Trypan blue to count the number of cells. Cell density was adjusted to 5 X which was injected into each mouse by 100 a0. 6 8 week old BALB/c mice (Charles River Japan, Inc.) were used. After the right side of abdomen was surgically excised, cancer cell suspension was carefully injected to the spleen using a needle. LK8 protein was subcutaneously injected to a experimental group twice a day by 10 mg/kg /day, and likewise, saline was injected as a control group. 14 days later, the mice were sacrificed to examine the liver (FIG. 3a), and the number of colonies seen on surface of the liver was counted, which was fixed in 10% formalin solution. There was no big difference in weight among groups. However, the number of colonies which were transferred to liver was much less in an experimental group which was administered with LK8 protein by 10 mg/kg/day than in a control group. The decrease of the number of colonies in an experimental group injected with LK8 protein by mg/kg/day was about 60%, comparing to a control group (FIG. 3b). Besides, liver tissue section WO 2004/073730 PCT/KR2004/000357 which was fixed in formalin solution was stained with H&E, and observed. As a result, a tumor region was much limited in an experimental group treated with LK8 protein comparing to a control group (FIG. 3c).
Example 4: Growth inhibition of a primary tumor In order to investigate the effect of LK8 protein on the anti-angiogenesis in vivo, a xenografted tumor model was used. Every relevant experiment was performed with 4 week old randomly crossed female Balb/c nu/nu nude mice (Charles River Japan, Inc.), which were raised under sterilized condition.
Human prostatic carcinoma (PC-3) Human prostatic carcinoma PC-3 cells (American Type Culture Collection) were cultured in RPMI 1640 medium (GIBCO
TM
Invitrogen Corporation) supplemented with 10% FBS, and about X 10 6 PC-3 cells were subcutaneously injected in central muscular region of the back of the nude mouse. Exactly 10 days after the transplantation, LK8 protein was injected by 100 mg/kg/day.
21 WO 2004/073730 PCT/KR2004/000357 Meanwhile, a control group was injected PBS only instead of LK8 protein. The treatment was continued for 30 days, then, the size of a tumor was measured every 3 or 4 days. As a result, tumor growth was inhibited by the treatment of LK8 protein, which was about 60% inhibition comparing to a control group (FIG. 4a). When the LK8 protein was injected by 50 mg/kg/day, tumor growth was inhibited in similar fashion over comparing to a control group (FIG. 4b).
Human lung carcinoma (A549) Human lung carcinoma A549 cells (American Type Culture Collection) were cultured in DMEM medium (GIBCO
T
Invitrogen Corporation) supplemented with 10% FBS. Then, 1 X 10 7 tumor cells were subcutaneously injected in central muscular region of the back of the nude mouse.
Exactly 5 days after the transplantation, LK8 protein was injected by 50 mg/kg/day. A control group was administered PBS only instead of LK8 protein. The treatment was continued for 46 days, then, the size of a tumor was measured every 3 or 4 days. As a result, tumor growth was inhibited by the treatment of LK8 protein, which was 61% 22 WO 2004/073730 PCT/KR2004/000357 inhibition comparing to a control (FIG. Human colon and rectal carcinoma (LS 174T) Human colon and rectal carcinoma LS 174T cells (American Type Culture Collection) were cultured in RPMI medium (GIBCOTM, Invitrogen Corporation) supplemented with 10% FBS. Then, X 106 tumor cells were subcutaneously injected in central muscular region of the back of the nude mouse. Exactly 5 days after the transplantation, LK8 protein was injected by 50 mg/kg/day. A control group was administered PBS only instead of LK8 protein. The treatment was continued for 34 days, then, the size of a tumor was measured every 3 or 4 days. As a result, tumor growth was inhibited by the treatment of LK8 protein, which was 64% inhibition comparing to a control (FIG.
6a). And, the weight of the tumor was decreased by the treatment of LK8 protein about 68.7%, which was measured on the final day of the experiment (FIG. 6b).
Example 5: Acute toxicity test of LK8 protein old SPF SD (Sprague Dawley) line rats 23 WO 2004/073730 PCT/KR2004/000357 were used in the test for acute toxicity. Rats were divided into 5 groups, and 5 rats per each group were administered once with LK8 protein by the dosage of 260 mg/kg, 364 mg/kg, 510 mg/kg, 714 mg /kg and 1000 -mg/kg, respectively, by intravenous injection (Table For 14 days after test material, LK8 protein, administration, death, clinical symptoms and weight change in rats were observed. The hematological tests and biochemical tests of blood were performed, and any abnormal signs in the gastrointestinal organs of chest and abdomen were examined visually during autopsy. As a result, a weak toxicity was detected in the group administered with 1000 mg/kg of LK8 protein, but neither toxicity nor death was found in other groups in most tests including weight changes, blood test, hematological tests and biochemical tests of blood, autopsy, etc. Therefore, the LK8 protein used in this experiment was evaluated to be safe substance since it did not cause any toxic change in rats up to the level of 1,000 mg/kg in rats and its estimated LD 50 values were much greater than 1,000 mg/kg in rats (Table 2) <Table 2> WO 2004/073730 PCT/KR2004/000357 The number of deaths according to days after LK8 protein administration Amou Days after LK 8 protein nt The administration of number 0 1 2 3 4 5 6 7 8 9 1 1 1 1 1 admi of 0 1 2 3 4 nist deaths rati /Total on test (ng/ rats kg) 260 0/5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 364 0/5 0 0 0 0 0 0 00 0 0 0 0 0 0 0 510 0/5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 714 0/5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1000 3/5 0 0 3 0 0 0 0 0 0 0 0 0 0 0 0 INDUSTRIAL APPLICABILITY As explained hereinbefore, LK8 protein has an inhibiting effect on metastasis, in particular, on the growth of human prostatic cancer, lung cancer, colon cancer and rectal cancer as being systemically administered. Thus, an anticancer agent containing LK8 protein of the present invention can be effectively used as a treatment agent for a primary tumor or a metastasis WO 2004/073730 PCT/KR2004/000357 inhibitor.
Those skilled in the art will appreciate that the conceptions and specific embodiments disclosed in the foregoing description may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present invention. Those skilled in the art will also appreciate that such equivalent embodiments do not depart from the spirit and scope of the invention as set forth in the appended claims.
Claims (5)
- 3. The pharmaceutical composition as set forth in claim 2, wherein the effective dose of LK8 protein is 1 50 mg/kg.
- 4. A pharmaceutical compostion when used to inhibit metastasis of cancer comprising LK8 protein represented by SEQ ID NO:1 as an effective ingredient. The pharmaceutical composition as set forth in claim 1, wherein the cancer is selected from the group consisting of colon cancer, melanoma and rectal cancer.
- 6. The pharmaceutical composition as set forth in claim 4, the metastasis is a metastasis of melanoma into lung or a metastasis of colon or rectal cancer into liver.
- 7. Use of LK8 protein represented by SEQ ID NO:1 in the manufacture of a medicament for treating a cancer selected from the group consisting of prostate cancer, colon cancer, rectal cancer and lung cancer or inhibiting a metastasis of said cancer.
- 8. Use as set forth in claim 7, wherein the metastasis is a metastasis of melanoma into lung or a metastasis of colon or rectal cancer into liver. W ftgeR750000 799999I753039\753039 Claims Jun O7 o
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020030010797A KR100595364B1 (en) | 2003-02-20 | 2003-02-20 | Anticancer agent comprising LK8 protein as an active ingredient |
KR10-2003-0010797 | 2003-02-20 | ||
PCT/KR2004/000357 WO2004073730A1 (en) | 2003-02-20 | 2004-02-20 | Anticancer agent comprising lk8 protein as an active ingredient |
Publications (2)
Publication Number | Publication Date |
---|---|
AU2004212856A1 AU2004212856A1 (en) | 2004-09-02 |
AU2004212856B2 true AU2004212856B2 (en) | 2007-07-19 |
Family
ID=36116045
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2004212856A Ceased AU2004212856B2 (en) | 2003-02-20 | 2004-02-20 | Anticancer agent comprising LK8 protein as an active ingredient |
Country Status (9)
Country | Link |
---|---|
EP (1) | EP1608395A4 (en) |
JP (1) | JP2006518342A (en) |
KR (1) | KR100595364B1 (en) |
CN (1) | CN100546646C (en) |
AU (1) | AU2004212856B2 (en) |
BR (1) | BRPI0407611A (en) |
CA (1) | CA2516172A1 (en) |
RU (1) | RU2306147C2 (en) |
WO (1) | WO2004073730A1 (en) |
Families Citing this family (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7285277B2 (en) | 2002-03-15 | 2007-10-23 | Mogam Biotechnology Research Institute | Anticancer agent |
CN1761680A (en) | 2002-12-26 | 2006-04-19 | 武田药品工业株式会社 | Metastin derivative and use thereof |
KR100681762B1 (en) * | 2004-01-09 | 2007-02-15 | 재단법인 목암생명공학연구소 | 68 8 Therapeutic agent for treatment of cancer comprising human apolipoprotein a kringles LK68 or LK8 genes as effective ingredient and method for treating cancer using the same |
CN101863971A (en) | 2004-06-25 | 2010-10-20 | 武田药品工业株式会社 | Metastin derivatives and use thereof |
TWI386417B (en) | 2005-12-22 | 2013-02-21 | Takeda Pharmaceutical | Metastin derivatives and use thereof |
US8404643B2 (en) | 2005-12-22 | 2013-03-26 | Takeda Pharmaceutical Company Limited | Metastin derivatives and use thereof |
TWI404726B (en) | 2006-10-25 | 2013-08-11 | Takeda Pharmaceutical | Metastin derivatives and use thereof |
KR100888022B1 (en) * | 2006-12-21 | 2009-03-09 | 재단법인 목암생명공학연구소 | Fusion Proteion of Imunoglobulin Fc and Human Apolipoproteina Kringle Fragment |
FR2919061B1 (en) | 2007-07-19 | 2009-10-02 | Biomerieux Sa | METHOD OF DOSING PLASTINE-I FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER. |
FR2919063B1 (en) | 2007-07-19 | 2009-10-02 | Biomerieux Sa | METHOD OF DETERMINING LEUCOCYTE ELASTASE INHIBITOR FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER. |
CA2693098C (en) | 2007-07-19 | 2019-06-18 | Yasemin Ataman-Onal | Dosage method for the carcino-embryonic antigen and the carbohydrate 19-9 antigen to hepatic fatty acid-binding protein for the diagnosis of colorectal cancer |
FR2919064B1 (en) * | 2007-07-19 | 2009-10-02 | Biomerieux Sa | METHOD OF ASSAYING APOLIPOPROTEIN ALL FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER |
FR2919065B1 (en) | 2007-07-19 | 2009-10-02 | Biomerieux Sa | METHOD FOR DETERMINING APOLIPOPROTEIN AI FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER |
FR2919062B1 (en) | 2007-07-19 | 2009-10-02 | Biomerieux Sa | METHOD OF DETERMINING AMINOACYLASE 1 FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER. |
FR2919060B1 (en) | 2007-07-19 | 2012-11-30 | Biomerieux Sa | METHOD OF DETERMINING EZRINE FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER |
FR2933773B1 (en) | 2008-07-10 | 2013-02-15 | Biomerieux Sa | METHOD FOR DETERMINING THE ISOMERASE DISULFIDE PROTEIN FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER |
WO2012067427A2 (en) * | 2010-11-16 | 2012-05-24 | 재단법인 목암생명공학연구소 | Pharmaceutical composition containing lk8 protein as an active ingredient for preventing or treating diabetic retinopathy or age-related macular degeneration |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001019868A1 (en) * | 1999-09-15 | 2001-03-22 | Mogam Biotechnology Research Institute | A novel angiogenesis inhibitor |
-
2003
- 2003-02-20 KR KR1020030010797A patent/KR100595364B1/en not_active IP Right Cessation
-
2004
- 2004-02-20 CA CA002516172A patent/CA2516172A1/en not_active Abandoned
- 2004-02-20 EP EP04713257A patent/EP1608395A4/en not_active Withdrawn
- 2004-02-20 CN CNB2004800048895A patent/CN100546646C/en not_active Expired - Fee Related
- 2004-02-20 WO PCT/KR2004/000357 patent/WO2004073730A1/en active IP Right Grant
- 2004-02-20 BR BRPI0407611-7A patent/BRPI0407611A/en not_active IP Right Cessation
- 2004-02-20 JP JP2006500650A patent/JP2006518342A/en not_active Withdrawn
- 2004-02-20 AU AU2004212856A patent/AU2004212856B2/en not_active Ceased
- 2004-02-20 RU RU2005129273/15A patent/RU2306147C2/en not_active IP Right Cessation
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001019868A1 (en) * | 1999-09-15 | 2001-03-22 | Mogam Biotechnology Research Institute | A novel angiogenesis inhibitor |
Also Published As
Publication number | Publication date |
---|---|
JP2006518342A (en) | 2006-08-10 |
CN100546646C (en) | 2009-10-07 |
KR100595364B1 (en) | 2006-07-03 |
CN1753685A (en) | 2006-03-29 |
RU2005129273A (en) | 2006-03-10 |
EP1608395A4 (en) | 2007-10-31 |
AU2004212856A1 (en) | 2004-09-02 |
RU2306147C2 (en) | 2007-09-20 |
WO2004073730A1 (en) | 2004-09-02 |
CA2516172A1 (en) | 2004-09-02 |
BRPI0407611A (en) | 2006-02-14 |
KR20040075270A (en) | 2004-08-27 |
EP1608395A1 (en) | 2005-12-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2004212856B2 (en) | Anticancer agent comprising LK8 protein as an active ingredient | |
Lin et al. | Astragaloside IV alleviates doxorubicin induced cardiomyopathy by inhibiting NADPH oxidase derived oxidative stress | |
Chowdhury et al. | Ultrasound-guided therapeutic modulation of hepatocellular carcinoma using complementary microRNAs | |
TWI362941B (en) | Methods for effecting regression of tumoer mass and size in a metastasized tumor | |
CN1252689A (en) | Combined tumor suppressor gene therapy and chemotherapy in treatment of neoplasms | |
EP3701949A1 (en) | Pharmaceutcal composition for preventing or treating cancer, containing streptonigrin and rapamycin as active ingredients | |
Kelland | Targeting established tumor vasculature: a novel approach to cancer treatment | |
EP4378453A1 (en) | Dual-targeting biomimetic liposome containing elemene and cabazitaxel, and preparation method therefor and use thereof | |
Vesely et al. | Novel therapeutic approach for cancer using four cardiovascular hormones | |
Wang et al. | A multifunctional non-viral vector for the delivery of MTH1-targeted CRISPR/Cas9 system for non-small cell lung cancer therapy | |
TWI547277B (en) | Honokiol for the treatment or prevention of bladder cancer growth and metastasis and improve the cachexia new use | |
HU229628B1 (en) | Angiogenesis and vascular permeability modulators and inhibitors | |
Kim et al. | Inhibitory effect of the salmosin gene transferred by cationic liposomes on the progression of B16BL6 tumors | |
Yao et al. | Tumor oxygenation nanoliposome synergistic hypoxia-inducible-factor-1 inhibitor enhanced Iodine-125 seed brachytherapy for esophageal cancer | |
EP1031352B1 (en) | Use of desmopressin in the preparation of a metastatic dissemination inhibitor medicament during cancer surgery | |
US7285277B2 (en) | Anticancer agent | |
CN1334738A (en) | Cerebrovascular regeneration/reconstruction promoters and nerve tissue secondary degeneration inhibitors comprising ginsenoside Rb1 | |
JPS61111693A (en) | An expression vector containing a λP▲L subscript▼ promoter and a restriction site engineered to conveniently replace the ribosome binding site, a plasmid containing this vector, and a plasmid containing this plasmid. Host, and related methods | |
Chen et al. | Endothelial progenitor cells combined with cytosine deaminase-endostatin for suppression of liver carcinoma | |
Luo et al. | A liposome-based combination strategy using doxorubicin and a PI3K inhibitor efficiently inhibits pre-metastatic initiation by acting on both tumor cells and tumor-associated macrophages | |
Chen et al. | An oncolytic system produces oxygen selectively in pancreatic tumor cells to alleviate hypoxia and improve immune activation | |
JPH10511677A (en) | Use of inositol triphosphate for drug preparation. | |
JP2003527337A (en) | Administration method of therapeutic agent based on anti-angiogenic schedule | |
JPWO2006035515A1 (en) | Pharmaceutical composition for treating or preventing superficial bladder cancer, and use thereof | |
Wu et al. | Caffeic acid phenethyl ester (cape) mediated decrease in metastasis of colon cancer cells: an in vitro and in vivo study |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
DA3 | Amendments made section 104 |
Free format text: THE NATURE OF THE AMENDMENT IS: DELETE CO-INVENTOR KIM, WON KYOUNG. ADD CO-INVENTOR PARK, JUNG HWAN |
|
FGA | Letters patent sealed or granted (standard patent) | ||
MK14 | Patent ceased section 143(a) (annual fees not paid) or expired |