CN104837859B - 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 - Google Patents
细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 Download PDFInfo
- Publication number
- CN104837859B CN104837859B CN201380057740.2A CN201380057740A CN104837859B CN 104837859 B CN104837859 B CN 104837859B CN 201380057740 A CN201380057740 A CN 201380057740A CN 104837859 B CN104837859 B CN 104837859B
- Authority
- CN
- China
- Prior art keywords
- peptide
- cell
- application
- active constituent
- composition
- 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.)
- Active
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 247
- 239000000203 mixture Substances 0.000 title claims abstract description 57
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 title abstract description 25
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 title abstract description 25
- 239000000470 constituent Substances 0.000 claims abstract description 48
- 210000004027 cell Anatomy 0.000 claims description 99
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 43
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 38
- 150000001413 amino acids Chemical class 0.000 claims description 31
- 108090000623 proteins and genes Proteins 0.000 claims description 30
- 102000004169 proteins and genes Human genes 0.000 claims description 28
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 26
- 102000039446 nucleic acids Human genes 0.000 claims description 23
- 108020004707 nucleic acids Proteins 0.000 claims description 23
- 201000010099 disease Diseases 0.000 claims description 22
- 150000007523 nucleic acids Chemical class 0.000 claims description 21
- 239000002872 contrast media Substances 0.000 claims description 18
- 239000003814 drug Substances 0.000 claims description 18
- 239000002537 cosmetic Substances 0.000 claims description 15
- 229940079593 drug Drugs 0.000 claims description 15
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 239000004615 ingredient Substances 0.000 claims description 10
- 239000008194 pharmaceutical composition Substances 0.000 claims description 9
- 150000001875 compounds Chemical class 0.000 claims description 8
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 6
- 230000021615 conjugation Effects 0.000 claims description 6
- 239000002105 nanoparticle Substances 0.000 claims description 5
- 230000005298 paramagnetic effect Effects 0.000 claims description 4
- 101000655352 Homo sapiens Telomerase reverse transcriptase Proteins 0.000 claims description 3
- 235000013402 health food Nutrition 0.000 claims description 3
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 3
- 229910052742 iron Inorganic materials 0.000 claims description 3
- 239000003446 ligand Substances 0.000 claims description 3
- 150000002632 lipids Chemical class 0.000 claims description 3
- 239000011707 mineral Substances 0.000 claims description 3
- 238000002601 radiography Methods 0.000 claims description 3
- 210000000130 stem cell Anatomy 0.000 claims description 3
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 2
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 claims 1
- 235000021060 food property Nutrition 0.000 claims 1
- 239000000047 product Substances 0.000 claims 1
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 42
- 239000000562 conjugate Substances 0.000 description 38
- 229940024606 amino acid Drugs 0.000 description 36
- 235000001014 amino acid Nutrition 0.000 description 36
- 238000000034 method Methods 0.000 description 31
- 235000018102 proteins Nutrition 0.000 description 26
- 239000000126 substance Substances 0.000 description 25
- 239000000463 material Substances 0.000 description 20
- 230000000694 effects Effects 0.000 description 19
- 239000003550 marker Substances 0.000 description 18
- 108020004414 DNA Proteins 0.000 description 17
- -1 aromatic amino acid Chemical class 0.000 description 17
- 238000006243 chemical reaction Methods 0.000 description 16
- 239000011347 resin Substances 0.000 description 16
- 229920005989 resin Polymers 0.000 description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 239000002253 acid Substances 0.000 description 15
- 230000008859 change Effects 0.000 description 15
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 14
- 230000003834 intracellular effect Effects 0.000 description 14
- 230000032258 transport Effects 0.000 description 13
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 11
- 239000004472 Lysine Substances 0.000 description 11
- 235000013339 cereals Nutrition 0.000 description 11
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 10
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 10
- 241000700605 Viruses Species 0.000 description 10
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 9
- 208000024891 symptom Diseases 0.000 description 9
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 8
- 235000004279 alanine Nutrition 0.000 description 8
- 210000004899 c-terminal region Anatomy 0.000 description 8
- 230000007547 defect Effects 0.000 description 8
- 210000003470 mitochondria Anatomy 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 7
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 7
- 239000004473 Threonine Substances 0.000 description 7
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 7
- 230000001580 bacterial effect Effects 0.000 description 7
- 235000013922 glutamic acid Nutrition 0.000 description 7
- 239000004220 glutamic acid Substances 0.000 description 7
- 230000013595 glycosylation Effects 0.000 description 7
- 238000006206 glycosylation reaction Methods 0.000 description 7
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 6
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 6
- 229910021529 ammonia Inorganic materials 0.000 description 6
- 235000009582 asparagine Nutrition 0.000 description 6
- 229960001230 asparagine Drugs 0.000 description 6
- 238000004113 cell culture Methods 0.000 description 6
- 238000012377 drug delivery Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000013604 expression vector Substances 0.000 description 6
- 238000000684 flow cytometry Methods 0.000 description 6
- 208000015181 infectious disease Diseases 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000007758 minimum essential medium Substances 0.000 description 6
- 239000013612 plasmid Substances 0.000 description 6
- 108091033319 polynucleotide Proteins 0.000 description 6
- 102000040430 polynucleotide Human genes 0.000 description 6
- 239000002157 polynucleotide Substances 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 6
- 102000055501 telomere Human genes 0.000 description 6
- 108091035539 telomere Proteins 0.000 description 6
- 210000003411 telomere Anatomy 0.000 description 6
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 6
- 238000012384 transportation and delivery Methods 0.000 description 6
- 239000004475 Arginine Substances 0.000 description 5
- 239000004471 Glycine Substances 0.000 description 5
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 5
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 5
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 5
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 5
- 229930182555 Penicillin Natural products 0.000 description 5
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 5
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 5
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 5
- 235000003704 aspartic acid Nutrition 0.000 description 5
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 230000004700 cellular uptake Effects 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 229940049954 penicillin Drugs 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 238000010926 purge Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 208000011580 syndromic disease Diseases 0.000 description 5
- 239000004474 valine Substances 0.000 description 5
- 208000019932 Aciduria Diseases 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 241000725303 Human immunodeficiency virus Species 0.000 description 4
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 4
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 4
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 4
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 4
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 description 4
- 108010088535 Pep-1 peptide Proteins 0.000 description 4
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 230000007812 deficiency Effects 0.000 description 4
- 208000035475 disorder Diseases 0.000 description 4
- 239000012894 fetal calf serum Substances 0.000 description 4
- 239000005090 green fluorescent protein Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 4
- 229960000310 isoleucine Drugs 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- 238000010647 peptide synthesis reaction Methods 0.000 description 4
- 229920001184 polypeptide Polymers 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 208000010444 Acidosis Diseases 0.000 description 3
- 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 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 3
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 3
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 3
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 3
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 3
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- 108010017842 Telomerase Proteins 0.000 description 3
- 150000001720 carbohydrates Chemical group 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000006071 cream Substances 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000006073 displacement reaction Methods 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 206010015037 epilepsy Diseases 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 3
- WTFXARWRTYJXII-UHFFFAOYSA-N iron(2+);iron(3+);oxygen(2-) Chemical compound [O-2].[O-2].[O-2].[O-2].[Fe+2].[Fe+3].[Fe+3] WTFXARWRTYJXII-UHFFFAOYSA-N 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 3
- 238000002953 preparative HPLC Methods 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 229940054269 sodium pyruvate Drugs 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000007790 solid phase Substances 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
- JHALWMSZGCVVEM-UHFFFAOYSA-N 2-[4,7-bis(carboxymethyl)-1,4,7-triazonan-1-yl]acetic acid Chemical compound OC(=O)CN1CCN(CC(O)=O)CCN(CC(O)=O)CC1 JHALWMSZGCVVEM-UHFFFAOYSA-N 0.000 description 2
- HBBSDZXXUIHKJE-UHFFFAOYSA-N 6-hydrazinylpyridine-3-carboxylic acid Chemical compound NNC1=CC=C(C(O)=O)C=N1 HBBSDZXXUIHKJE-UHFFFAOYSA-N 0.000 description 2
- 101710103970 ADP,ATP carrier protein Proteins 0.000 description 2
- 101710133192 ADP,ATP carrier protein, mitochondrial Proteins 0.000 description 2
- 208000006096 Attention Deficit Disorder with Hyperactivity Diseases 0.000 description 2
- 208000036864 Attention deficit/hyperactivity disease Diseases 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 208000031229 Cardiomyopathies Diseases 0.000 description 2
- 108020004635 Complementary DNA Proteins 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 101710088194 Dehydrogenase Proteins 0.000 description 2
- 102100024746 Dihydrofolate reductase Human genes 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 2
- 206010048804 Kearns-Sayre syndrome Diseases 0.000 description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 201000000639 Leber hereditary optic neuropathy Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- 208000007014 Retinitis pigmentosa Diseases 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- 108020004459 Small interfering RNA Proteins 0.000 description 2
- 108091060271 Small temporal RNA Proteins 0.000 description 2
- 241001655322 Streptomycetales Species 0.000 description 2
- 208000006011 Stroke Diseases 0.000 description 2
- 102000019259 Succinate Dehydrogenase Human genes 0.000 description 2
- 108010012901 Succinate Dehydrogenase Proteins 0.000 description 2
- KKEYFWRCBNTPAC-UHFFFAOYSA-N Terephthalic acid Chemical compound OC(=O)C1=CC=C(C(O)=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-N 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 102100026383 Vasopressin-neurophysin 2-copeptin Human genes 0.000 description 2
- LBAFWCXLYPHUPY-UHFFFAOYSA-N acetic acid;n-(2-aminoethyl)-n-benzylhydroxylamine Chemical compound CC(O)=O.CC(O)=O.NCCN(O)CC1=CC=CC=C1 LBAFWCXLYPHUPY-UHFFFAOYSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- YVPYQUNUQOZFHG-UHFFFAOYSA-N amidotrizoic acid Chemical compound CC(=O)NC1=C(I)C(NC(C)=O)=C(I)C(C(O)=O)=C1I YVPYQUNUQOZFHG-UHFFFAOYSA-N 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- TZCXTZWJZNENPQ-UHFFFAOYSA-L barium sulfate Chemical compound [Ba+2].[O-]S([O-])(=O)=O TZCXTZWJZNENPQ-UHFFFAOYSA-L 0.000 description 2
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 2
- 230000002146 bilateral effect Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000010804 cDNA synthesis Methods 0.000 description 2
- 230000032823 cell division Effects 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000002591 computed tomography Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 201000010064 diabetes insipidus Diseases 0.000 description 2
- 229960005423 diatrizoate Drugs 0.000 description 2
- 108020001096 dihydrofolate reductase Proteins 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 238000004520 electroporation Methods 0.000 description 2
- 239000002532 enzyme inhibitor Substances 0.000 description 2
- 229940125532 enzyme inhibitor Drugs 0.000 description 2
- 239000003797 essential amino acid Substances 0.000 description 2
- 235000020776 essential amino acid Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- KTWOOEGAPBSYNW-UHFFFAOYSA-N ferrocene Chemical compound [Fe+2].C=1C=C[CH-]C=1.C=1C=C[CH-]C=1 KTWOOEGAPBSYNW-UHFFFAOYSA-N 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- IZOOGPBRAOKZFK-UHFFFAOYSA-K gadopentetate Chemical compound [Gd+3].OC(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O IZOOGPBRAOKZFK-UHFFFAOYSA-K 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 235000001497 healthy food Nutrition 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 125000001841 imino group Chemical group [H]N=* 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 208000003531 maternally-inherited Leigh syndrome Diseases 0.000 description 2
- 108091070501 miRNA Proteins 0.000 description 2
- 239000002679 microRNA Substances 0.000 description 2
- 230000002438 mitochondrial effect Effects 0.000 description 2
- 230000008811 mitochondrial respiratory chain Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 239000000863 peptide conjugate Substances 0.000 description 2
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 150000003053 piperidines Chemical class 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 229920000139 polyethylene terephthalate Polymers 0.000 description 2
- 239000005020 polyethylene terephthalate Substances 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000000750 progressive effect Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000000241 respiratory effect Effects 0.000 description 2
- 230000035939 shock Effects 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 229910001936 tantalum oxide Inorganic materials 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 108091006106 transcriptional activators Proteins 0.000 description 2
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 2
- 229940045145 uridine Drugs 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- SWZCTMTWRHEBIN-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(4-hydroxyphenyl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=C(O)C=C1 SWZCTMTWRHEBIN-QFIPXVFZSA-N 0.000 description 1
- JZTKZVJMSCONAK-INIZCTEOSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-hydroxypropanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CO)C(O)=O)C3=CC=CC=C3C2=C1 JZTKZVJMSCONAK-INIZCTEOSA-N 0.000 description 1
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- IZKGGDFLLNVXNZ-KRWDZBQOSA-N (2s)-5-amino-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-oxopentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCC(=O)N)C(O)=O)C3=CC=CC=C3C2=C1 IZKGGDFLLNVXNZ-KRWDZBQOSA-N 0.000 description 1
- KYBXNPIASYUWLN-WUCPZUCCSA-N (2s)-5-hydroxypyrrolidine-2-carboxylic acid Chemical compound OC1CC[C@@H](C(O)=O)N1 KYBXNPIASYUWLN-WUCPZUCCSA-N 0.000 description 1
- DHBXNPKRAUYBTH-UHFFFAOYSA-N 1,1-ethanedithiol Chemical compound CC(S)S DHBXNPKRAUYBTH-UHFFFAOYSA-N 0.000 description 1
- HSQUQVDLRZNEQQ-UHFFFAOYSA-N 1,3,7,10-tetrazacyclotetradec-9-ene Chemical compound N1=CCNCCCNCNCCCC1 HSQUQVDLRZNEQQ-UHFFFAOYSA-N 0.000 description 1
- HFDVRLIODXPAHB-UHFFFAOYSA-N 1-tetradecene Chemical compound CCCCCCCCCCCCC=C HFDVRLIODXPAHB-UHFFFAOYSA-N 0.000 description 1
- SLROPVABSCVNOL-UHFFFAOYSA-N 2,2,4,6,7-pentamethyl-3a,4-dihydro-3h-1-benzofuran Chemical class CC1C=C(C)C(C)=C2OC(C)(C)CC12 SLROPVABSCVNOL-UHFFFAOYSA-N 0.000 description 1
- MGHMWKZOLAAOTD-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(1h-indol-3-yl)propanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)NC(C(=O)O)CC1=CNC2=CC=CC=C12 MGHMWKZOLAAOTD-UHFFFAOYSA-N 0.000 description 1
- GRUVVLWKPGIYEG-UHFFFAOYSA-N 2-[2-[carboxymethyl-[(2-hydroxyphenyl)methyl]amino]ethyl-[(2-hydroxyphenyl)methyl]amino]acetic acid Chemical compound C=1C=CC=C(O)C=1CN(CC(=O)O)CCN(CC(O)=O)CC1=CC=CC=C1O GRUVVLWKPGIYEG-UHFFFAOYSA-N 0.000 description 1
- HHLZCENAOIROSL-UHFFFAOYSA-N 2-[4,7-bis(carboxymethyl)-1,4,7,10-tetrazacyclododec-1-yl]acetic acid Chemical compound OC(=O)CN1CCNCCN(CC(O)=O)CCN(CC(O)=O)CC1 HHLZCENAOIROSL-UHFFFAOYSA-N 0.000 description 1
- UXIVXGFGVAWZCH-UHFFFAOYSA-N 2h-triazepine Chemical compound N1N=CC=CC=N1 UXIVXGFGVAWZCH-UHFFFAOYSA-N 0.000 description 1
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- SLXKOJJOQWFEFD-UHFFFAOYSA-N 6-aminohexanoic acid Chemical compound NCCCCCC(O)=O SLXKOJJOQWFEFD-UHFFFAOYSA-N 0.000 description 1
- 208000013824 Acidemia Diseases 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- 201000003695 Alexia Diseases 0.000 description 1
- 241000752021 Alexia Species 0.000 description 1
- 206010003591 Ataxia Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 241000700199 Cavia porcellus Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 206010008190 Cerebrovascular accident Diseases 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 201000000915 Chronic Progressive External Ophthalmoplegia Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241000218691 Cupressaceae Species 0.000 description 1
- 102000000634 Cytochrome c oxidase subunit IV Human genes 0.000 description 1
- 108090000365 Cytochrome-c oxidases Proteins 0.000 description 1
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 229910052692 Dysprosium Inorganic materials 0.000 description 1
- 102000015782 Electron Transport Complex III Human genes 0.000 description 1
- 108010024882 Electron Transport Complex III Proteins 0.000 description 1
- 229910052691 Erbium Inorganic materials 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 229910052693 Europium Inorganic materials 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000287828 Gallus gallus Species 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
- 229930186217 Glycolipid Natural products 0.000 description 1
- 208000032087 Hereditary Leber Optic Atrophy Diseases 0.000 description 1
- 208000034604 Hereditary acrokeratotic poikiloderma Diseases 0.000 description 1
- 241000175212 Herpesvirales Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 108010070875 Human Immunodeficiency Virus tat Gene Products Proteins 0.000 description 1
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 1
- 206010020674 Hypermetabolism Diseases 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 description 1
- 208000035177 MELAS Diseases 0.000 description 1
- 208000035172 MERRF Diseases 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- 108020005196 Mitochondrial DNA Proteins 0.000 description 1
- 206010051403 Mitochondrial DNA deletion Diseases 0.000 description 1
- 208000035155 Mitochondrial DNA-associated Leigh syndrome Diseases 0.000 description 1
- 206010058799 Mitochondrial encephalomyopathy Diseases 0.000 description 1
- 201000002169 Mitochondrial myopathy Diseases 0.000 description 1
- 208000013300 Multiple mitochondrial DNA deletion syndrome Diseases 0.000 description 1
- 208000010428 Muscle Weakness Diseases 0.000 description 1
- 208000021642 Muscular disease Diseases 0.000 description 1
- 206010028372 Muscular weakness Diseases 0.000 description 1
- 102000006386 Myelin Proteins Human genes 0.000 description 1
- 108010083674 Myelin Proteins Proteins 0.000 description 1
- 208000036572 Myoclonic epilepsy Diseases 0.000 description 1
- 208000002033 Myoclonus Diseases 0.000 description 1
- 201000009623 Myopathy Diseases 0.000 description 1
- OKIZCWYLBDKLSU-UHFFFAOYSA-M N,N,N-Trimethylmethanaminium chloride Chemical compound [Cl-].C[N+](C)(C)C OKIZCWYLBDKLSU-UHFFFAOYSA-M 0.000 description 1
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 1
- 230000004988 N-glycosylation Effects 0.000 description 1
- 241000244206 Nematoda Species 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 1
- 102000019040 Nuclear Antigens Human genes 0.000 description 1
- 108010051791 Nuclear Antigens Proteins 0.000 description 1
- 206010061323 Optic neuropathy Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 208000013234 Pearson syndrome Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- 101710149951 Protein Tat Proteins 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 1
- 101100240886 Rattus norvegicus Nptx2 gene Proteins 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 108091027967 Small hairpin RNA Proteins 0.000 description 1
- 239000004902 Softening Agent Substances 0.000 description 1
- 241000399119 Spio Species 0.000 description 1
- 208000000389 T-cell leukemia Diseases 0.000 description 1
- 208000028530 T-cell lymphoblastic leukemia/lymphoma Diseases 0.000 description 1
- 101150006914 TRP1 gene Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- WDLRUFUQRNWCPK-UHFFFAOYSA-N Tetraxetan Chemical compound OC(=O)CN1CCN(CC(O)=O)CCN(CC(O)=O)CCN(CC(O)=O)CC1 WDLRUFUQRNWCPK-UHFFFAOYSA-N 0.000 description 1
- 241000710914 Totivirus Species 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- CWERGRDVMFNCDR-UHFFFAOYSA-N alpha-mercaptoacetic acid Natural products OC(=O)CS CWERGRDVMFNCDR-UHFFFAOYSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 1
- 230000001195 anabolic effect Effects 0.000 description 1
- 230000001166 anti-perspirative effect Effects 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 239000003213 antiperspirant Substances 0.000 description 1
- 201000007201 aphasia Diseases 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 125000000613 asparagine group Chemical group N[C@@H](CC(N)=O)C(=O)* 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 208000015802 attention deficit-hyperactivity disease Diseases 0.000 description 1
- 150000003851 azoles Chemical class 0.000 description 1
- 159000000009 barium salts Chemical class 0.000 description 1
- QRUDEWIWKLJBPS-UHFFFAOYSA-N benzotriazole Chemical compound C1=CC=C2N[N][N]C2=C1 QRUDEWIWKLJBPS-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 102000005936 beta-Galactosidase Human genes 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000021523 carboxylation Effects 0.000 description 1
- 238000006473 carboxylation reaction Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 201000006662 cervical adenocarcinoma Diseases 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- 231100000244 chromosomal damage Toxicity 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 150000004700 cobalt complex Chemical class 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000002826 coolant Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 239000007822 coupling agent Substances 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 108010082025 cyan fluorescent protein Proteins 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- GPRSOIDYHMXAGW-UHFFFAOYSA-N cyclopenta-1,3-diene cyclopentanecarboxylic acid iron Chemical compound [CH-]1[CH-][CH-][C-]([CH-]1)C(=O)O.[CH-]1C=CC=C1.[Fe] GPRSOIDYHMXAGW-UHFFFAOYSA-N 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 230000020176 deacylation Effects 0.000 description 1
- 238000005947 deacylation reaction Methods 0.000 description 1
- 230000006240 deamidation Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 150000001991 dicarboxylic acids Chemical class 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000006196 drop Substances 0.000 description 1
- 206010013932 dyslexia Diseases 0.000 description 1
- KBQHZAAAGSGFKK-UHFFFAOYSA-N dysprosium atom Chemical compound [Dy] KBQHZAAAGSGFKK-UHFFFAOYSA-N 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000001037 epileptic effect Effects 0.000 description 1
- UYAHIZSMUZPPFV-UHFFFAOYSA-N erbium Chemical compound [Er] UYAHIZSMUZPPFV-UHFFFAOYSA-N 0.000 description 1
- UKFXDFUAPNAMPJ-UHFFFAOYSA-N ethylmalonic acid Chemical compound CCC(C(O)=O)C(O)=O UKFXDFUAPNAMPJ-UHFFFAOYSA-N 0.000 description 1
- OGPBJKLSAFTDLK-UHFFFAOYSA-N europium atom Chemical compound [Eu] OGPBJKLSAFTDLK-UHFFFAOYSA-N 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
- 230000007661 gastrointestinal function Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 235000015220 hamburgers Nutrition 0.000 description 1
- 239000007902 hard capsule Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 239000003668 hormone analog Substances 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 150000002466 imines Chemical class 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000023692 inborn mitochondrial myopathy Diseases 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000001524 infective effect Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000001023 inorganic pigment Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- ICIWUVCWSCSTAQ-UHFFFAOYSA-M iodate Chemical compound [O-]I(=O)=O ICIWUVCWSCSTAQ-UHFFFAOYSA-M 0.000 description 1
- 150000002497 iodine compounds Chemical class 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 150000003951 lactams Chemical class 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 230000005291 magnetic effect Effects 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 208000028260 mitochondrial inheritance Diseases 0.000 description 1
- 230000023202 mitochondrion inheritance Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 210000005012 myelin Anatomy 0.000 description 1
- 230000007498 myristoylation Effects 0.000 description 1
- 229940031182 nanoparticles iron oxide Drugs 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 210000001577 neostriatum Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000001272 neurogenic effect Effects 0.000 description 1
- 230000002232 neuromuscular Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 229910000069 nitrogen hydride Inorganic materials 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 208000020911 optic nerve disease Diseases 0.000 description 1
- 239000012860 organic pigment Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- BPUBBGLMJRNUCC-UHFFFAOYSA-N oxygen(2-);tantalum(5+) Chemical compound [O-2].[O-2].[O-2].[O-2].[O-2].[Ta+5].[Ta+5] BPUBBGLMJRNUCC-UHFFFAOYSA-N 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 230000026792 palmitoylation Effects 0.000 description 1
- 208000024691 pancreas disease Diseases 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000006072 paste Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- BRBAEHHXGZRCBK-UHFFFAOYSA-N pentrinitrol Chemical compound [O-][N+](=O)OCC(CO)(CO[N+]([O-])=O)CO[N+]([O-])=O BRBAEHHXGZRCBK-UHFFFAOYSA-N 0.000 description 1
- 229950006286 pentrinitrol Drugs 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920003216 poly(methylphenylsiloxane) Polymers 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 150000003214 pyranose derivatives Chemical class 0.000 description 1
- 229940076788 pyruvate Drugs 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012106 screening analysis Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 239000002884 skin cream Substances 0.000 description 1
- 239000004055 small Interfering RNA Substances 0.000 description 1
- RBJZIQZDAZLXEK-UHFFFAOYSA-M sodium;3-hydroxy-2-methylpropanoate Chemical compound [Na+].OCC(C)C([O-])=O RBJZIQZDAZLXEK-UHFFFAOYSA-M 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 108010004034 stable plasma protein solution Proteins 0.000 description 1
- 230000037351 starvation Effects 0.000 description 1
- 208000023516 stroke disease Diseases 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- PXQLVRUNWNTZOS-UHFFFAOYSA-N sulfanyl Chemical class [SH] PXQLVRUNWNTZOS-UHFFFAOYSA-N 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- GUVRBAGPIYLISA-UHFFFAOYSA-N tantalum atom Chemical compound [Ta] GUVRBAGPIYLISA-UHFFFAOYSA-N 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- ZERJNAGZUCPHNB-UHFFFAOYSA-N tetrazane Chemical compound NNNN ZERJNAGZUCPHNB-UHFFFAOYSA-N 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000002054 transplantation Methods 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
- 210000005239 tubule Anatomy 0.000 description 1
- 229910052721 tungsten Inorganic materials 0.000 description 1
- 239000010937 tungsten Substances 0.000 description 1
- UDKYUQZDRMRDOR-UHFFFAOYSA-N tungsten Chemical compound [W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W][W] UDKYUQZDRMRDOR-UHFFFAOYSA-N 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1276—RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
- A61K49/0043—Fluorescein, used in vivo
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0056—Peptides, proteins, polyamino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/57—Compounds covalently linked to a(n inert) carrier molecule, e.g. conjugates, pro-fragrances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/04—X-ray contrast preparations
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/10—Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2458/00—Labels used in chemical analysis of biological material
- G01N2458/30—Electrochemically active labels
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Wood Science & Technology (AREA)
- Birds (AREA)
- Gastroenterology & Hepatology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Dermatology (AREA)
- General Engineering & Computer Science (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Pharmacology & Pharmacy (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
Abstract
本发明公开了细胞穿透肽、作为活性成分的缀合物及其应用。具体而言,本发明公开了具有SEQ ID NO:1至SEQ ID NO:156中一种序列的细胞穿透肽、包含SEQ IDNO:1至SEQ ID NO:156中一种序列的片段的肽的缀合物、或包含与所述肽序列具有至少80%同源性的肽的缀合物、和包含所述缀合物的组合物。
Description
技术领域
本发明涉及源自人端粒酶逆转录酶(human telomerase reversetranscriptase;hTERT)的细胞穿透肽、细胞穿透肽及活性成分的缀合物、以及包含该缀合物的组合物。
背景技术
尽管低分子量物质、核酸、蛋白质、纳米粒子等在分子水平上具有作为治疗物质的极大可能性潜力,但低分子量物质、核酸、蛋白质、粒子等的使用其应用由于不能无能力穿透组织及细胞膜而受限。输送此类物质至细胞内的的系统的开发展已成为过去二十年来活跃的研究领域。细胞内部运输物质已成为分子处理治疗方法中的的话题。低分子量物质、核酸或纳米粒子通过系通过多种若干试剂、电穿孔或热休克而运送到运输到细胞内。然而,难以找到一种适当方法在不破坏蛋白质的活性及完整性的情况下将蛋白质输送到在细胞内部输送蛋白质。在20世纪80年代,在对人类人免疫缺乏病毒(human immunodeficiencyvirus;HIV)的细胞穿透能力进行的研究中,已发现,由特定的11种个氨基酸构组成的的HIV-TAT蛋白质在于在向细胞内部的运输的过程中起重要作用。因此,在20世纪90年代,探索运输到细胞内蛋白质将蛋白质运输到细胞内的恰当方法的研究成为重点研究领域。
已知端粒是染色体末端发现的遗传物质重复序列,端粒防止染色体损伤或防止合并至其它染色体上。端粒的长度在每次细胞分裂时缩短,且在一定次数的细胞分裂的后,端粒长度极度缩短至细胞停止分裂且死亡的程度。在另一方面,已知端粒延伸可延长细胞的寿命。作为实例,癌细胞分泌称为端粒酶的酶,该酶防止端粒缩短,因此导致癌细胞增殖。
本发明的目标在于提供一种新型肽。
本发明的另一目标在于提供编码该新型肽的多核苷酸。
本发明的另一目标在于提供细胞穿透肽。
本发明的另一目标在于提供作为细胞内活性成分的载体的有用的肽。
本发明的另一目标在于提供活性成分与细胞穿透肽缀合成的缀合物。
本发明的另一目标在于提供包含活性成分与细胞穿透肽的缀合物的组合物。
本发明的另一目标在于提供包含活性成分与细胞穿透肽的缀合物的药物组合物。
本发明的另一目标在于提供包含活性成分与细胞穿透肽的缀合物的功能性化妆组合物。
本发明的另一目标在于提供包含活性成分与细胞穿透肽的缀合物的健康食品组合物。
本发明的另一目标在于提供包含活性成分与细胞穿透肽的缀合物的对比造影剂。
发明内容
根据本发明的一个实施方式的缀合物可为细胞穿透载体肽与活性成分的缀合物,其中所述载体肽为包含SEQ ID NO:1至SEQ ID NO:156中至少一个氨基酸序列的肽、与上述序列具有至少80%同源性的肽、或上述肽的片段,且其中具有至少80%同源性的所述肽和所述片段保留SEQ ID NO:1至SEQ ID NO:156中的任一氨基酸序列的细胞穿透能力。
根据本发明中的缀合物的另一实施方式,所述片段可由3个或更多氨基酸构成。
根据本发明中的缀合物的另一实施方式,所述载体肽可由30个或更少氨基酸构成。
根据本发明中的缀合物的另一实施方式,上述载体肽可为由SEQ ID NO:1至SEQID NO:156中任一氨基酸序列构成的肽。
根据本发明中的缀合物的另一实施方式,其包含选自以下组的任一氨基酸序列,所述组由SEQ ID NO:9、SEQ ID NO:37、SEQ ID NO:43、SEQ ID NO:48、SEQ ID NO:52、SEQID NO:55、SEQ ID NO:90、SEQ ID NO:92和SEQ ID NO:121组成。
根据本发明的一个实施方式的对比造影剂可包含上述的任一种缀合物。
根据本发明的一个实施方式的对比造影剂可用于细胞对比造影。
根据本发明的对比造影剂的另一实施方式,所述细胞可为干细胞。
根据本发明的一个实施方式的组合物可包含上述的任一种缀合物。
根据本发明的组合物的另一实施方式,所述活性成分可用于治疗或预防疾病,且所述组合物可为药物组合物。
根据本发明的组合物的另一实施方式,所述活性成分可为用于功能性化妆品的活性成分,且所述组合物可为化妆品组合物。
根据本发明的组合物的另一实施方式,所述活性成分可为用于功能健康食品的活性成分,且所述组合物可为健康食品组合物。
根据本发明方法的一个实施方式,所述方法可为用于输送活性成分至细胞内的方法,其中该方法包括对有需要的对象施用上述任何缀合物,且其中载体肽为将活性成分输送至细胞内的细胞穿透肽,且其中具有至少80%同源性的所述肽及上述肽的片段保留由SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列构成的肽的细胞穿透能力。
根据本发明的方法的另一实施方式,所述方法可用于将所述活性成分局部地输送至细胞内部的粒线体内。
根据本发明细胞穿透肽的另一实施方式,上述载体肽可以为具有SEQ ID NO:1至SEQ ID NO:156中任意一个或多个氨基酸序列的肽。
根据本发明的多核苷酸可编码上述细胞穿透肽。
根据本发明的载体可包含上述多核苷酸。
根据本发明的转化细胞可包含上述载体。
产业利用性
难以运输到细胞内的活性成分可通过使用本发明所公开的肽或肽及活性成分的缀合物而易于运输到细胞内。这意味着可增加活性成分的功效且因此可减少活性成分的剂量。因此,可最小化由于药物施用造成的副作用且可增加治疗的有效性。特别地,当将药物局部地输送至粒线体内时,可改善粒线体相关的疾病或病症,且可增加疾病预防及治疗疾病的有效性。在化妆品的情况中,通过少量活性成分可产生显著效果。通过使肽与对比造影物质缀合,可将该肽用作对比造影物质以监测细胞移植的过程或在细胞治疗中的移植细胞。特别地,该肽可实际上用作用于注入身体内的干细胞的对比造影物质。
附图说明
图1至图23描绘通过FACS分析的在FITC与SEQ ID No:1至SEQ ID No:156的肽Pep1融合后治疗的HeLa细胞中细胞摄入的细胞数量。对照细胞仅以FITC处理。
图24至图43描绘通过FACS分析的在FITC与SEQ ID NO:1至SEQ ID NO:156的肽融合后治疗的Huh7细胞中细胞摄入的细胞数量。对照细胞仅以FITC处理。
图44至图58描绘通过FACS分析的在FITC与SEQ ID NO:1至SEQ ID NO:156的肽融合后治疗的人T淋巴球细胞系(Jurkat细胞)中细胞摄入的细胞数量。对照细胞仅以FITC处理。
图59至图72描绘通过FACS(流式细胞术)分析的在FITC与SEQ ID No:1至SEQ IDNo:156的肽融合后治疗的HeLa细胞的毒性及细胞活性的结果。对照细胞仅以FITC处理。
具体实施方式
尽管蛋白质、核酸、肽或病毒等具有作为治疗物质的极大可能性潜力,但蛋白质、核酸、肽或病毒等的使用由于无能力不能穿透组织及细胞膜而受限。即使分子大小很小,但该等这种分子由于分子的结构或特性而无法穿透脂质双层。因此,试图经由通过使用电穿孔、热休克等将蛋白质、核酸、肽或病毒运输到细胞内;难以在既不破坏细胞膜又保持上述分子的活性状态的情况下转移彼等所述蛋白质、核酸、肽或病毒。已进行的许多研究显示源自人免疫缺乏病毒(Human Immuno-deficiency Virus;HIV)的反式转录活化因子(Trans-Activating Transcriptional activator;TAT activator)蛋白质可用作细胞穿透肽,该细胞穿透肽可将巨极大的活性物质到运输细胞内。具体地,已进行了关于下列物质的研究,与在细胞内部产生毒性的TAT蛋白质不同,该等该物质可在不产生任何毒性的情况下运输诸如蛋白质、核酸、肽或病毒的极巨大分子到细胞内。因此,本发明是通过本发明人发现源自端粒酶的肽具有作为细胞穿透肽的显著功效而无显著明显毒性而完成的。
肽在下表1中所示的SEQ ID NO:1至SEQ ID NO:156公开。SEQ ID NO:157是人端粒酶蛋白质的全长序列。下表1中的“名称”系用于区别肽。在本发明的不同特定实施方式中,在SEQ ID NO:1至SEQ ID NO:156中超过一个肽包括“合成肽”,即,端粒酶的选定区域的合成肽。在本说明书中,术语“pep”在本文系指具有SEQ ID NO:1至SEQ ID NO:156中任一氨基序列者的肽、包含与上述序列具有至少80%同源性的氨基酸序列的肽或上述肽的片段。
[表1]
在本发明的一个实施方式中,一种多核苷酸编码以下的肽:包含SEQ ID NO:1至SEQ ID NO:156中至少一个氨基酸序列的肽,与上述序列具有至少80%同源性的肽或作为上述肽的片段的肽。如上所述的多核苷酸能够使得所述肽大量地产生。举例而言,培养包括编码肽的多核苷酸的载体使得肽大量产生。
本文公开的肽可包括包含同源性超过80%、超过85%、超过90%、超过95%、超过96%、超过97%、超过98%、超过99%的氨基酸序列的肽。此外,本发明中所公开的肽可包括:包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列的肽或该肽的片段,及具有超过1个转化(transformed)氨基酸、超过2个转化氨基酸、超过3个转化氨基酸、超过4个转化氨基酸、超过5个转化氨基酸、超过6个转化氨基酸或超过7个转化氨基酸的肽。
在本发明的一个实施方式中,氨基酸序列中的改变属于肽的物理及化学特性的变化。举例而言,氨基酸转化可经执行而用于改善肽的热稳定性、改变底物特异性及改变最佳pH值。
术语“氨基酸”在本文不仅包括经天然引入到肽中的22种标准氨基酸,还包括D-异构体及转化氨基酸。因此,在本发明的特定实施方式中,本文的肽包括具有D-氨基酸的肽。在另一方面,肽可包括非标准氨基酸,诸如已经翻译后修饰的那些氨基酸。翻译后修饰的实例包括磷酸化、糖基化、酰化(包括乙酰化、豆蔻酰化、棕榈酰化)、烷化、羧化、羟化、糖化、生物素化、泛素化、化学性质的转化(例如,β-移除脱酰亚胺、脱酰胺)及结构转化(例如,形成双硫键)。同样,包括氨基酸的改变,氨基酸由于在为形成肽缀合物而与交联剂组合过程中发生化学反应而发生改变。
本文公开的肽可为已经识别且从天然源分离出的野生型肽。在另一方面,当与SEQID NO:1至SEQ ID NO:156中任一氨基酸序列的肽片段相比时,本文公开的肽可为人工突变株突变体,该人工突变株包含一个或多个经取代、删除缺失及/或嵌入添加的一或更多个氨基酸。在野生型多肽中变化(不仅在人工突变株中)的氨基酸变化包含不显著影响活性蛋白质折迭折叠及/或活性的氨基酸的保守取代。保守取代的实例属于由碱性氨基酸(精氨酸、赖氨酸及组氨酸)、酸性氨基酸(谷氨酸及天冬氨酸)、极性氨基酸(谷酰胺及天冬酰胺)、疏水性氨基酸(亮氨酸、异亮氨酸、缬氨酸及甲硫氨酸)、芳香族氨基酸(苯丙氨酸、色氨酸及酪氨酸)及小型氨基酸(甘氨酸、丙氨酸、丝氨酸及苏氨酸)组成的组。通常不改变特定活性的氨基酸取代在本技术领域中已知。最常发生的变化为丙氨酸/丝氨酸、缬氨酸/异亮氨酸、天冬氨酸/谷氨酸、苏氨酸/丝氨酸、丙氨酸/甘氨酸、丙氨酸/苏氨酸、丝氨酸/天冬酰胺、丙氨酸/缬氨酸、丝氨酸/甘氨酸、酪氨酸/苯丙氨酸、丙氨酸/脯氨酸、赖氨酸/精氨酸、天冬氨酸/天冬酰胺、亮氨酸/异亮氨酸、亮氨酸/缬氨酸、丙氨酸/谷氨酸、天冬氨酸/甘氨酸,及相反的变化。保守取代的另一实例显示在下表2中。
[表2]
肽的生物学性质的实质转化通过选择下列功效方面的显著不同的置换而执行:(a)保持置换区域中的多肽主链结构的功效,诸如片状或三维螺旋结构,(b)保持靶区域中分子的电荷或疏水性的功效,或(c)保持侧链的整体的功效。自然残基根据一般侧链性质划分成如下组:
(1)疏水性:正亮氨酸、甲硫氨酸、丙氨酸、缬氨酸、亮氨酸、异亮氨酸;
(2)中性亲水性:半胱氨酸、丝氨酸、苏氨酸;
(3)酸性:天冬氨酸、谷氨酸;
(4)碱性:天冬酰胺、谷氨酰胺、组氨酸、赖氨酸、精氨酸;
(5)影响链取向(chain orientation)的残基:甘氨酸、脯氨酸;和
(6)芳香性:色氨酸、酪氨酸、苯丙氨酸。
非保守置换可通过将上述种类的成员更换为不同种类的成员而进行。与保持肽的适当三维结构无关的任何半胱氨酸残基可通常经置换为丝氨酸,因此增加分子的氧化稳定性且防止不适当交联。反之,稳定性的改善可通过给肽添加一个或更多个半胱氨酸键而实现。
肽的氨基酸变体的经改变类型为已改变抗体糖基化方式的那些氨基酸。术语「改变」在本文是指删除在肽中发现的至少一个糖残基和/或添加在肽内不存在的至少一个糖基化残基。
肽中的糖基化作用通常经N连接或O连接。术语“N-连接”在本文是指将糖残基附着至天冬酰胺残基的侧链。作为三肽序列,天冬酰胺-X-丝氨酸和天冬酰胺-X-苏氨酸(其中X为除脯氨酸的外的任何氨基酸)为用于将糖残基酶法附着至天冬酰胺的侧链的识别序列。因此,在多肽中存在该三肽序列中的一种的情况下,创建可能的糖基化作用位点。“O-连接糖基化作用”意味着将糖N-乙酰半乳胺糖、半乳糖或木糖中的一种附着至羟基氨基酸。羟基氨基酸大部分通常为丝氨酸或苏氨酸,但也可使用5-羟基脯氨酸或5-羟基赖氨酸。
对肽添加糖基化作用位点通过改变氨基酸序列为含有上述三肽序列(用于N连结的糖基化作用位点)而便利地执行。该改变可通过给第一抗体序列添加至少一个丝氨酸或苏氨酸残基或通过以那些残基(用于O连结的糖基化作用位点)置换而进行。
在本发明的一个实施方式中,提供包含肽的细胞穿透肽,其中该肽包含SEQ IDNO:1至SEQ ID NO:156中任一氨基酸序列,该肽具有与上述序列超过80%同源性的氨基酸序列,或肽为上述肽的片段。
在本发明的一个实施方式中,提供药物组合物,该药物组合物包含肽作为运送一个以上活性成分的药物递送系统,其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,该肽具有与上述序列超过80%的同源性,或该肽为上述肽的片段。
包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列的肽、上述肽的片段或与上述序列具有超过80%同源性的肽是安全的且具有作为细胞穿透肽的显著功效。因此,肽可与药物缀合以在细胞内部运送药物。
在本发明的一个实施方式中,提供肽与待运送的活性成分的缀合物,其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,该肽为上述肽的片段或该肽与上述肽具有超过80%的同源性。在本发明的一个实施方式中,活性成分可为选自以下物质中的至少一种:蛋白质、核酸、肽、脂质、醣脂质、矿物质、糖、对比造影物质、药物和化合物。在本发明的一个实施方式中,活性成分可为肽。在本发明的一个实施方式中,活性成分可为细胞因子、抗体、抗体片段、治疗性酶、可溶性受体或配体。
本文揭示的细胞穿透肽意指可自体外和/或体内运送货物(cargo)至细胞内部的肽。本文揭示的「货物」包含可通过与细胞穿透肽的缀合作用在细胞内部运送的所有物质,例如,想要增加细胞穿透功效的所有物质,具体而言,药物、化妆品或健康食品的活性成分,更具体而言,无法通过一般途径在细胞内部运送的物质,更具体而言,糖、纳米颗粒、生物制剂、病毒、对比造影物质或其它化合物,该其它化合物可例如具有蛋白质、核酸、肽、矿物质、葡萄糖,但不限于那些物质。本文揭示的“药物”为宽泛概念,包括待运送用于缓和、预防、治疗或诊断疾病、创伤或特定症状的物质。
本文揭示的“载体肽”为可通过与活性成分的缀合作用运送活性成分至靶位点的肽。
在本发明的一个实施方式中,作为货物的蛋白质或肽包含以下的一或更多者:激素、激素类似物、酶、酶抑制剂、信号转移蛋白质(或肽)、抗体和疫苗,但不限于那些物质。在本发明的一个实施方式中,核酸为以下分子:可为自发或人工的、单链或双链的DNA分子或RNA分子。核酸分子可为相同类型(例如,具有相同的核苷酸序列)的一或更多个核酸或不同类型的核酸。核酸分子包含以下的一或更多者:DNA、互补DNA(cDNA)、诱饵DNA(decoy DNA)、RNA、小干扰RNA(siRNA)、微小RNA(miRNA)、小发夹式RNA(shRNA)、小时序RNA(stRNA)、小核仁RNA(snoRNA)、小胞核RNA(snRNA)、戊糖核酸(pentose nucleic acid;PNA)、反义寡聚物、质粒和其它修饰核酸,但不限于那些物质。在本发明的一个实施方式中,病毒包含全病毒或包括病毒的核酸的病毒核心。在本发明的一个实施方式中,化学物质为包含天然或合成物质的宽泛指示,该天然或合成物质可充当药物。
在本发明的一个实施方式中,可通过藉由细胞穿透肽运送到在细胞内的部运输之药物是可随包含一或更多个药物递送的运输载体,诸如脂质体、胶束微胞、奈米纳米粒子、磁性粒子或量子点。
本文揭示的术语“对比造影物质”为宽泛指示,该宽泛指示包含用来在医学影像中对比躯体内的结构或流体的所有物质。适当对比造影物质包含不透射线对比造影剂、顺磁对比造影剂、超顺磁对比造影剂、计算机断层扫描(computed tomography;CT)和其它对比造影物质,但不限于那些物质。举例而言,不透射线对比造影剂(用于X射线影像)将包含无机碘化合物和有机碘化合物(例如,泛影酸盐(diatrizoate))、不透射线金属和该不透射线金属的盐(例如,银、金、铂等)和其它不透射线化合物(例如,钙盐、诸如硫酸钡的钡盐、钽和氧化钽)。适当的顺磁对比造影物质(用于MR影像)包含钆二乙三胺五乙酸(gadoliniumdiethylene triaminepentaacetic acid;Gd-DTPA)和Gd-DTPA的衍生物、其它钆、锰、铁、镝、铜、铕、铒、铬、镍和钴复合体,例如,1,4,7,10-四氮杂环十二烷-N,N’,N”,N”’-四乙酸(1,4,7,10-tetraazacyclododecan-N,N’,N”,N”’-tetraacetic acid;DOTA)、乙二胺四乙酸(ethylenediaminetetraacetic acid;EDTA)、1,4,7,10-四氮杂环十二烷-N,-N’,N”-三乙酸(1,4,7,10-tetraazacyclododecan-N,-N’,N”-triacetic acid;DO3A)、1,4,7-三氮杂环壬烷-N,N’,N”-三乙酸(1,4,7-triazacyclononane-N,N’,N”-TRIACETIC ACID;NOTA)、1,4,8,10-四氮杂环十四烷-N,N’,N”,N”’-四乙酸(1,4,8,10-tetraazacyclotetradecane-N,N’,N”,N”’-tetraacetic acid;TETA)、羟基苄基乙二胺二乙酸(hydroxybenzylethylene-diamine diacetic acid;HBED)。适当的超顺磁对比造影物质(用于MR影像)包含磁铁矿、超顺磁氧化铁(super-paramagnetic iron oxide;SPIO)、超小超顺磁氧化铁(ultrasmallsuperparamagnetic iron oxide;USPIO)和单晶氧化铁。其它适当的对比造影物质为碘化、非碘化、离子和非离子的CT对比造影剂、类似旋转标记或诊断上有效制剂的对比造影物质。
对比造影物质的其它实例包含β-半乳糖苷酶、绿色荧光蛋白、青色荧光蛋白、荧光素酶(但不限于那些物质)和编码在表示于细胞内时可容易检测到的蛋白质的标识基因。可使用各种标记,诸如放射性同位素、flour、酶、酶基质、酶辅因子、酶抑制剂、配体(尤其是半抗原(heptan))。
在本发明的一实例中,对比造影物质为下文化学式2的二茂铁羧酸。二茂铁的结构系表示在化学式1中。
在本发明的一个实例中,细胞穿透肽与对比造影物质的缀合物为下文化学式3中表示的二茂铁羧基-pep。
在本发明的一个实施方式中,肽或组合物可与一个或更多个可检测标记融合。标记可为可在化学反应、物理反应或酶反应中检测到的化合物或在反应中直接或间接地产生信号的化合物。标记和检测随后可根据此技术领域所熟知的方法来执行(例如,Sambrook,J.和Russel,D.W.(2001);和Lottspeich,F.和Zorbas H.(1998)Bioanalytik,SpektrumAkademischer Verlag,海德堡/柏林,德国)。标记包含荧光标记、酶标记、显色标记、发光标记、辐射标记、半抗原、生物素、金属复合体、金属和胶体金,但不限于那些标记。该标记的所有形式在此工作领域是众所周知的,该标记的所有形式可自各供货商购得。
在本发明的一个实施方式中,货物可与肽直接结合。在本发明的另一个实施方式中,货物可通过诸如共价键或非共价键的各种类型的键与肽结合。例如,在本发明的一个实施方式中,货物可与肽的N端或C端结合。举例而言,货物可通过二硫键或共价键键结至肽。共价键为可将货物键结至N端谷氨酸的α-胺或C端赖氨酸残基的胺的键。同样,肽和货物可通过非共价键结合,此可使肽或货物可将彼此封装为胶囊形式。
在本发明的另一个实施方式中,肽可通过接头与货物结合。举例而言,肽可通过在将诸如联肼尼克酰胺(6-肼基吡啶-3-羧酸)接头(Hynic(6-hydrazinopyridine-3-carboxylic acid)linker)的接头引入N端谷氨酸的α-胺或C端赖氨酸残基的胺之后将货物结合至接头而与货物结合。
在本发明的另一个实施方式中,当货物为DNA或RNA时,将SH基团(硫醇基)引入肽,且将马来酰亚胺基引入DNA或RNA,随后,肽的SH基团和DNA或RNA的马来酰亚胺基结合,因此产生在货物与肽之间的结合。
在本发明的另一个实施方式中,当货物为肽或蛋白质时,表达货物的DNA与表达载体肽的DNA结合,且通过表达该DNA结合,可将货物与肽结合为融合蛋白质的形式。通过融合蛋白质结合的特定实例如下:当制造引物用于产生融合蛋白质时,编码载体肽的核苷酸经附着在表达货物的核苷酸前面,且使用限制酶将所获得核苷酸嵌入诸如聚对苯二甲酸乙二醇酯(Polyethylene Terephthalate;pET)载体的载体,且核苷酸系以转化到诸如BL-21(DE3)的细胞中来表达。此时,融合蛋白质将通过以类似异丙基-1-硫代-β-D-半乳糖吡喃糖苷(isopropyl-1-thio-β-D-galactopyranoside;IPTG)的表达诱导制剂处理该融合蛋白质而有效表达。随后,所表达的融合蛋白质系通过His标签净化来净化,且以PBS透析,且经添加至试剂盒以在2000rpm至4000rpm、5至20分钟的此类条件下通过离心分离作用而浓缩。
在本发明的一个实施方式中,载体肽与染色物质、荧光物质、特别是异硫氰酸荧光素(fluorescein isothiocyanate;FITC)或绿色荧光蛋白(Green Fluorescent Protein;GFP)结合。在本发明的一个实施方式中,FITC系与在载体肽的N端或C端的赖氨酸的胺基(NH3+)结合。在其中赖氨酸不存在于肽的端的肽情况中,肽可通过包括赖氨酸的接头与FITC结合。
本文揭示的载体肽可以1:1的摩尔分数与货物结合,但该载体肽也可以不同于1:1的摩尔分数与货物结合,该载体肽为包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列的肽,或具有与上述肽超过80%同源性的氨基酸序列的肽,或上述肽的片段。举例而言,CPP与货物的摩尔分数可大于2:1,具体而言,大于2:1、大于3:1、大于4:1、大于5:1、大于6:1、大于7:1、大于8:1、大于9:1或大于10:1。此意味着大量载体肽分子可与货物分子结合。大量载体肽分子可经串行或并行结合。“串行结合”意味着载体肽与货物分子将在端氨基酸处结合。“并行结合”意味着载体肽与货物分子将在不同于端氨基酸的位点结合。在另一方面,载体肽与货物的摩尔分数可大于1:2。此意味着载体肽分子可与大量货物分子结合。举例而言,载体肽与货物的摩尔分数可为1:2,具体而言,大于1:2、大于1:3、大于1:4、大于1:5、大于1:6、大于1:7、大于1:8、大于1:9或大于1:10。
可轻易发现与异硫氰酸荧光素结合的肽的移动途径。因此,在本发明的一个实施方式中的载体肽将用于细胞成像或检测在细胞内部的药物递送途径。
在本发明的一个实施方式中,提供肽作为运送一个以上活性成分的药物递送载体的用途,其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,或该肽为上述肽的片段,或该肽具有与上述肽超过80%同源性的氨基酸序列。用途可指治疗用途或非治疗用途。
在本发明的一个实施方式中,提供在将药物递送到受试对象的细胞内部的方法,该方法包含施用包含药物和肽的组合物的步骤;其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,或该肽为上述肽的片段,或该肽具有与上述肽超过80%同源性的氨基酸序列。
在本发明的一个实施方式中,提供检测药物递送途径的方法,该方法包含对受试对象应用肽和对比造影物质的步骤;其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,或该肽为上述肽的片段,或该肽具有与上述肽超过80%同源性的氨基酸序列。
在本发明的一个实施方式中,提供检测药物递送途径的方法,该方法包含对受试对象应用肽与对比造影物质的缀合物的步骤;其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,或该肽为上述肽的片段,或该肽具有与上述肽超过80%同源性的氨基酸序列。
在本发明的一个实施方式中,提供用于将药物递送至受试对象的细胞内的试剂盒,该试剂盒含有组合物和说明书,其中该组合物包含本发明的肽与用于递送的药物的缀合物,其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列或该肽为上述肽的片段,或该肽具有与上述肽超过80%同源性的氨基酸序列,其中该说明书包括以下的至少一种:给药剂量、给药途径、给药频率和组合物的指示。
在本发明的一个实施方式中,提供包含活性成分和肽的化妆品或食品组合物;其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,该肽具有与上述序列超过80%同源性的氨基酸序列,或该肽为上述肽的片段。在本发明的另一个实施方式中,提供包含肽与活性成分的缀合物的化妆品或食品组合物;其中该肽包含SEQ ID NO:1至SEQ IDNO:156中任一氨基酸序列,该肽具有与上述序列超过80%同源性的氨基酸序列,或该肽为上述肽的片段。
在本发明的一个实施方式中,提供具有将活性成分运送到细胞内部的显著能力的药物、化妆品或食品组合物,该药物、化妆品或食品组合物包含肽与活性成分的缀合物;其中该肽包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列,该肽具有与上述序列超过80%同源性的氨基酸序列,或该肽为上述肽的片段。
线粒体,作为真核细胞的能量代谢中的中心胞器,为与人类疾病有关的首先熟知的细胞内胞器(Luft R、Ikkos D、Palmieri G、Ernster L、Afzelius B:A case of severehypermetabolism of non thyroid origin with a defect in the maintenance ofmitochondrial respiratory control:a correlated clinical,biochemical,andmorphological study(在保持线粒体呼吸控制中具有缺陷的非甲状腺成因的严重代谢亢进的情况:相关的临床研究、生物化学研究和形态学研究),J Clin Invest 41:1776-804,1962年)。
因为线粒体在控制细胞的能量代谢和细胞凋亡中起重要作用,故该线粒体充当各种治疗药物的主要靶。同样,此胞器涉和控制细胞内部的钙浓度,线粒体呼吸链充当在能量产生中重要的电子运送系统,且该线粒体呼吸链导致产生活性氧物种。因此,异常线粒体作用与成人疾病具有密切关系,该成人疾病诸如尿崩症、心肌症、不孕症、失明、肾/肝疾病和中风(Modica-Napolitano KS,Singh KK:四月mitochondri as targets for detectionand treatment of cancer.(作为检测和治疗癌症的靶的线粒体。)Expert Rev Mol Med11:1-19,2002年)。同样,正建议将线粒体遗传突变包括在老化、变性神经元疾病和癌症等的爆发中。
根据本发明一个实施方式提供的粒线体靶向输送系统可包含包含上述的任一种缀合物,其中载体肽局部地移动至粒线体内,并且执行将所述活性成分局部细胞内粒线体输送的作用,其中与上述序列具有至少80%同源性的氨基酸序列的肽及片段是保持线粒体靶向递送系统的肽,上述线粒体靶向肽可以是具有SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列的肽。
可提供粒线体活性调节组合物,其中该组合物包含本发明的肽和递送用载体肽的缀合物,其中载体肽局部地移动至细胞内粒线体内,并且执行将所述活性成分局部细胞内粒线体输送的作用,其中与上述序列具有至少80%同源性的氨基酸序列的肽及该肽的片段为是保持线粒体靶向递送系统的肽,上述线粒体靶向肽可以是具有SEQ ID NO:1至SEQ IDNO:156中任一氨基酸序列的组合物。
本发明一个实施方式的粒线体活性调节组合物,所述组合物作为药物组合物用于治疗与粒线体有关之疾病或病症,预防、抑制疾病进展,或缓解症状;其中所述活性成分用于治疗与粒线体有关之疾病或病症,预防、抑制疾病进展,或缓解症状。
本文揭示的“线粒体相关疾病”包含亨汀顿氏疾病、肌萎缩性脊髓侧索硬化、线粒体脑肌肉病变合并乳酸血症和类中风症候群(Mitochondrial Encephalomyopathy withLactic Acidemia and Stroke-like episodes;MELAS);肌阵挛癫痫合并红色褴褛肌纤维症(Myoclonus,epilepsy,and myopathy with ragged red fibers;MERRF);神经性肌肉松弛、失调症、色素性视网膜炎/母体遗传莱氏症状(Neurogenic muscular weakness,ataxia,retinitis pigmentosa/Maternally inherited leigh syndrome;NARP/MILS);Leber氏视神经病变(Lebers hereditary optic neuropathy;LHON);Kearns-Sayre症候群(Kearns-Sayre Syndrome;KSS);皮尔森骨髓胰腺症(Pearson Marrow-PancreasSyndrome;PMPS);慢性渐进性眼外肌麻痹(Chronic progressive externalopthalnoplegia;CPEO);瑞氏症候群;阿尔珀斯氏症候群;多个线粒体DNA缺失症候群;线粒体DNA耗乏症候群;复合体Ⅰ缺陷;复合体Ⅱ(琥珀酸脱氢酶(succinatedehydrogenase;SDH))缺陷;复合体Ⅲ缺陷;细胞色素c氧化酶(COX,复合体Ⅳ)缺陷;复合体Ⅴ缺陷;腺嘌呤核苷酸转运体(Adenine nucleotide translocator;ANT)缺陷;丙酮酸脱氢酶(Pyruvatedehydrogenase;PDH)缺陷;具有乳酸血症的乙基丙二酸酸性尿;具有乳酸血症的3-甲基戊烯二酸酸性尿;体现为在传染期间的衰减的不应性癫痫;体现为在传染期间的衰减的阿斯伯格症候群;体现为在传染期间的衰减的自闭症;注意力不足过动症(Attention deficithyperactivity disorder;ADHD);体现为在传染期间的衰减的脑性麻痹;体现为在传染期间的衰减的失读症;母系遗传血小板减少症;白血病;MNGIE(线粒体肌病变、周围和自主神经病变、胃肠功能异常和癫痫);MARIAHS症候群(线粒体失调症、复发传染、失语症、低尿酸血症/髓磷脂减少症(hypomyelination)、癫痫发作和二羧酸酸性尿);ND6肌张力不全症;体现为在传染期间的衰减的周期性呕吐症状;具有乳酸血症的3-羟基异丁酸酸性尿;具有乳酸血症的尿崩症;尿苷反应性神经症状(Uridine reactive neural syndrome;URNS);家族双侧纹状体坏死(Familial bilateral striatum necrosis;FBSN);与胺基糖苷有关的听力损失;松驰心肌病;脾淋巴瘤;钨症状;多个线粒体DNA缺失症状;和肾小管酸血症/尿崩症/失调症症状,但不限于那些疾病。
在本发明的另一个实施方式中,提供编码上述多肽的核酸分子。例如,核酸分子具有碱基序列GAA GCG CGC CCG GCG CTG CTG ACC AGC CGC CTG CGC TTT ATT CCG AAA(SEQNO.:181)。核酸可根据熟习此项技术者所熟知的方法引入寄主细胞内。举例而言,熟知方法可为通过以下方法的转化方法:磷酸钙方法、脂质粒、电穿孔、接触病毒和细胞,或直接微注射至细胞内等。寄主细胞为较高等的真核细胞(例如,哺乳动物细胞),或较低等的真核细胞(诸如酵母细胞),或原核细胞(诸如细菌细胞)。适于转化的原核寄主细胞可为属于以下的物种:例如,大肠杆菌、枯草杆菌、沙门氏菌、假单胞菌、链霉菌和微细菌物种。
包括上述核酸分子的载体通常为重组表达载体,且该载体包含赋能寄主细胞转化的复制起点和可选择标记物(例如,用于真核细胞培养的二氢叶酸还原酶,或新霉素的容许度、四环素或氨苄西林在大肠杆菌中的容许度,酵母菌TRP1基因),和用于控制蛋白质涂布序列的转录的启动子。例如,可用表达载体为:熟知细菌质粒,诸如SV40、pcDNA的衍生物;和熟知细菌质粒,诸如colE1、pCR1、pBR322、pMal-C2、pET、pGEX(Smith等人,Gene 67:31-40(1988));质粒,诸如pMB9和pMB9的衍生物RP4;与噬菌体I的大量衍生物相同的噬菌体DNA,诸如NM989;诸如M13和丝状单股噬菌体DNA的噬菌体DNA;酵母质粒,例如,噬菌体DNA或自使用表达抑制序列的修饰质粒与噬菌体DNA的组合诱导的载体。哺乳动物表达载体包含复制起点、适当启动子和增强子。同样,该载体可包含强制核糖体结合位点、聚腺苷酸化位点、剪接供体和受体部分、转录终止序列和5’侧接(planking)非转录序列。哺乳动物表达载体可包含可诱导启动子,例如,含有二氢叶酸还原酶启动子的载体,含有DHFR表达盒或DHFR/甲氨喋呤共扩增载体的任何表达载体,诸如pED(Randal J,kaufman,1991,RandalJ.Kaufman,Current Protocols in Molycular Biology,16,12(1991))。或者,可使用以下载体:谷氨酰胺合成酶/甲硫氨酸磺酰亚胺共扩增载体,例如,pEE14(Celltech公司)、人类疱疹病毒第四型(Epstein-Barr-Virus;EBV);或受核抗原(EBNA)控制引导附加型表达的载体,例如,pREP4(Invitrogen公司)、pCEP4(Invitrogen公司)、pMEP4(Invitrogen公司)、pREP8(Invitrogen公司)、pREP9(Invitrogen公司)和pEBVHis(Invitrogen公司)。可选择哺乳动物表达载体为Rc/CMV(Invitrogen公司)和pRc/RSV(Invitrogen公司)等。可用于本发明的牛痘病毒哺乳动物表达载体为pSC11、pMJ601、pTKgptF1S等。
将用于本发明的酵母表达载体系统为非融合pYES2载体(Invitrogen公司)、融合pYESHisA、B、C(Invitrogen公司)、pRS载体等。
上述载体可经引入各种细胞,诸如哺乳动物细胞(特别是源自人类的细胞)或细菌、酵母、真菌、昆虫、线虫和植物细胞。适当细胞的实例为:VERO细胞;HELA细胞,例如ATCCNo.CCL2;CHO细胞系,例如ATCC No.CCL61;COS细胞,例如COS-7细胞和ATCC No.CRL 1650细胞;W138、BHK、HepG2、3T3,例如,ATCC No.CRL6361;A549、PC12、K562细胞;293细胞;Sf9细胞,例如ATCC No.CRL1711;和Cv1细胞,诸如ATCC No.CCL70等。
将用于本发明的其它适当细胞为原核寄主细胞菌株,例如,属于大肠杆菌(例如,DH5-α菌株)、枯草杆菌、沙门氏菌、假单胞菌、链霉菌和葡萄球菌的菌株。
在本发明的一个实施方式中,组合物可含有0.1μg/mg至1mg/mg、具体而言1μg/mg至0.5mg/mg、更具体而言10μg/mg至0.1mg/mg的以下肽:包含SEQ ID NO:1至SEQ ID NO:156中任一氨基酸序列的肽、包含与上述序列具有超过80%同源性的氨基酸序列的肽或上述肽的片段。当所包含的肽在上述范围内时,组合物的所有安全性和稳定性可经满足且在成本有效性方面为适当的。
在本发明的一个实施方式中,组合物可应用于所有动物,包括人类、狗、鸡、猪、母牛、羊、天竺鼠和猴。
在本发明的一个实施方式中,药物组合物可在骨髓、硬膜外或皮下手段中通过以下方式给药:口服、直肠给药、经皮给药、静脉给药、肌肉给药、腹膜内给药。
口服给药的形式可为但不限于片剂、药丸、软胶囊或硬胶囊、颗粒、粉末、溶液或乳液。非口服给药的形式可为但不限于注射剂、滴剂、洗剂、软膏、凝胶、乳霜、悬浮液、水乳液、栓剂、贴剂或喷剂。
在本发明的一个实施方式中,若有必要,药物组合物可含有添加剂,诸如稀释剂、赋形剂、润滑剂、黏合剂、崩解剂、缓冲剂、分散剂、表面活化剂、着色剂、芳香剂或甜味剂。在本发明的一个实施方式中,药物组合物可通过此技术领域中的习知工业方法制造。
在本发明的一个实施方式中,医学组合物的活性成分可根据以下方面变化:病人的年龄、性别、体重、病理和状态、给药途径或开药者的判断。基于该因子的剂量系在熟习此项技术者的水平内决定,且每日剂量例如可为但不限于,0.1μg/kg/天至1g/kg/天,具体而言1μg/kg/天至10mg/kg/天,更具体而言10μg/kg/天至1mg/kg/天,更具体而言50μg/kg/天至100μg/kg/天。在本发明的一个实施方式中,药物组合物可每天一至三次给药,但不限于此。
在本发明的一个实施方式中,化妆品组合物可以适于局部应用的所有形式来提供。举例而言,该形式可经提供为溶液、通过油相在水中的分散获得的水乳液、通过水在油相中的分散获得的水乳液、悬浮液、固体、凝胶、粉末、糊剂、泡沫、或气溶胶。该形式可通过此技术领域中的习知工业方法制造。
在本发明的一个实施方式中,化妆品组合物可在不会损害主效应的水平内包括可合意地增加主效应的其它成分。在本发明的一个实施方式中,化妆品组合物可另外包括润肤膏、软化剂、表面活化剂、UV吸收剂、防腐剂、杀真菌剂、抗氧化剂、pH值调节剂、有机颜料或无机颜料、芳香剂、冷却剂或止汗剂。上述成分的配方比可在不会损害本发明的目的和效应的水平内通过熟习此项技术者决定,且基于化妆品组合物的总重量的配方比可为以重量计0.01%至5%,具体而言以重量计0.01%至3%。
在本发明的一个实施方式中,食品组合物不局限于形式,但例如可为颗粒、粉末、液体和固体形式。每一形式可以由熟习此项技术者适当选取的常用于工业中的成分(除活性成分外)组成,且可增加具有其它成分的效应。
对于上述活性成分的剂量的判定系在熟习此项技术者的水平内,且每日剂量例如可为1μg/kg/天至10mg/kg/天,更具体而言10μg/kg/天至1mg/kg/天,更具体而言50μg/kg/天至100μg/kg/天,但不限于该数量且可根据年龄、健康状态、并发病和其它各种因子而变化。
本文使用的术语意欲用来描述实施例,而非用来限制本发明。前文数不胜数的术语不欲限制量而是表示可存在一个以上的所使用术语的事物。术语“包括”、“具有”、“组成”和“包含”应为开放解释(亦即,“包括但不限于”)。
使用数量的范围的提及代替说明范围内的分开数量,因此除非明确说明,否则每一数量可读作本文整合的分开数量。所有范围的端值系包括在范围内且可经独立组合。
除非另作说明或明显同上下文相矛盾,否则本文提及的所有方法可按适当顺序执行。除非包括在申请专利范围内,否则任一个实施方式和所有实施例或示例性语言(例如,使用“类似......”的语言)的使用系用来更清楚描述本发明,而不是限制本发明的范畴。在申请专利范围外的本文任何语言将不会解释为本发明的必需品。除非另外界定,否则本文使用的技术术语和科学术语具有本发明所归属的熟习此项技术者通常理解的意义。
本发明的优选实施方式系为执行本发明的发明者所熟知的最佳模式。对熟习此项技术者而言,先于优选实施例中的变化而读取声明之后,可为清楚的。本发明者希望熟习此项技术者可充分使用该变化且以不同于本文所列举的其它方式进行本发明。因此,如由专利法所允许,本发明包括在随附申请专利范围中所说明的关键点的等效物和等效物的变化。另外,在上述组分的任何组合内的所有可能变化系包括在本发明中,除非另外明确说明或同上下文相矛盾。尽管本发明系通过示例性实施例描述和表示,但熟习此项技术者将很好理解,在不脱离本发明的精神和范围的情况下可存在通过下文申请专利范围所界定的形式和细节上的各种变化。
实施例1:肽的合成
具有SEQ ID NO:1至SEQ ID NO:156的肽系根据固相肽合成的现有方法来合成。详细地,肽通过使用ASP48S(Peptron公司,大田市,大韩民国)利用Fmoc固相肽合成(solidphase peptide synthesis;SPPS)自C端偶联每一氨基酸而合成。那些肽经使用如下,那些肽在C端的第一氨基酸附着至树脂:
NH2-赖氨酸(Boc)-2-氯-三苯甲基树脂
NH2-丙氨酸-2-氯-三苯甲基树脂
NH2-精氨酸(Pbf)-2-氯-三苯甲基树脂
合成肽的所有氨基酸材料在N端通过Fmoc保护,且氨基酸残基通过可溶于酸的Trt、Boc、t-Bu(t-丁酯)、Pbf(2,2,4,6,7-五甲基二氢苯并呋喃-5-磺酰基)保护。诸如:
Fmoc-丙氨酸-OH、Fmoc-精氨酸(Pbf)-OH、Fmoc-谷氨酸(OtBu)-OH、Fmoc-脯氨酸-OH、Fmoc-亮氨酸-OH、Fmoc-异亮氨酸-OH、Fmoc-苯丙氨酸-OH、Fmoc-丝氨酸(tBu)-OH、Fmoc-苏氨酸(tBu)-OH、Fmoc-赖氨酸(Boc)-OH、Fmoc-谷氨酰胺(Trt)-OH、Fmoc-色氨酸(Boc)-OH、Fmoc-蛋氨酸-OH、Fmoc-天冬酰胺(Trt)-OH、Fmoc-酪氨酸(tBu)-OH、Fmoc-氨基己酸-OH、Trt-巯乙酸。
HBTU[2-(1H-苯并三唑-1-基)-1,1,3,3-六氟磷酸四甲铵]/HOBt[N-羟基苯并三唑]/NMM[4-甲基吗啡啉]用作偶联试剂。使用含20%哌啶的二甲基甲酰胺(dimethylformamide;DMF)移除Fmoc。为自残基移除保护或使所合成肽与树脂分离,使用分裂混合物[三氟乙酸(trifluoroacetic acid;TFA)/三异丙基硅烷(triisopropylsilane;TIS)/乙二硫醇(ethanedithiol;EDT)/H2O=92.5/2.5/2.5/2.5]。
肽通过使用固相分子框架通过以如下顺序过程添加每一氨基酸而合成:氨基酸保护、偶联反应、洗涤和去保护。在自树脂切断所合成肽之后,所合成肽通过高效液相层析法(High Performance Liquid Chromatography;HPLC)净化且通过质谱测定法(massspectrometry;MS)验证合成且随后冷冻干燥。
特定肽合成过程以SEQ ID NO:164的pep1(EARPALLTSRLRFIPK)的实施例描述如下。
1)偶联
将以NH2-赖氨酸(Boc)-2-氯-三苯甲基树脂保护的氨基酸(8当量)熔融在偶联剂HBTU(8当量)/HOBt(8当量)/NMM(16当量)中,且在添加DMF之后,将反应混合物在室温下培育达2小时,随后以DMF、MeOH和DMF顺序洗涤。
2)Fmoc去保护
在DMF中添加20%哌啶之后,将反应混合物在室温下培育达5分钟2次,随后以DMF、MeOH和DMF顺序洗涤。
3)通过反复重复反应1和反应2构成肽的碱性框架。
4)切割:添加分裂混合物至完全合成的肽且使肽与树脂分离。
5)将预冷却乙醚添加至混合物内,且随后使反应混合物离心分离以沉淀出肽。
6)在通过Prep-HPLC净化之后,通过LC/MS检查分子重量且冷冻干燥以获得粉末形式的肽。
实施例2:pep(CPP)-FITC缀合物的制备
(1)FITC-CPP缀合物的制备
如下制造与FITC结合的具有SEQ ID NO:1至SEQ ID NO:156的肽的缀合物,例如,pep1(SEQ ID NO:157)与FITC的缀合物,换言之,FITC-接头-pep1经制造如下。
根据实施例1中描述的制造方法获得的肽的碱性框架NH2-接头-E(OtBu)-A-R(Pbf)-P-A-L-L-T(tBu)-S(tBu)-R(Pbf)L-R(Pbf)-F-I-P-K(Boc)-2-氯-三苯甲基树脂与FITC反应。具体而言,将荧光素-5-异硫氰酸酯(FITC)(8当量)与N,N-二异丙基乙胺(N,N-Diisopropylethylamine;DIPEA)(16当量)熔融在DMF中。添加DMF溶液且在室温下反应达2小时,随后以DMF、MeOH和DMF顺序洗涤。因此,获得FITC-接头-E(OtBu)-A-R(Pbf)-P-A-L-L-T(tBu)-S(tBu)-R(Pbf)L-R(Pbf)-F-I-P-K(Boc)-2-氯-三苯甲基树脂。本文的接头为6-胺基己酸(Ahx)。TFA/TIS/H2O=95/2.5/2.5经添加至在树脂上组成的肽,且缀合物系与树脂分离。预冷却乙醚经添加至所获得混合物,且使用离心分离作用来沉淀肽缀合物。在通过Prep-HPLC净化之后,纯度系以分析HPLC确定且分子重量系通过LC/MS决定。如上所述合成的肽系通过由LC/MS确定分子重量而验证为FITC-pep1。随后将缀合物冷冻干燥。按照对SEQID NO:157的pep1所述相同的方式,还制备了SEQ ID NO:1至SEQ ID NO:156的肽与FITC融合的缀合物。
(2)CPP-FITC缀合物的制备
根据实施例21.(1)中描述的制造方法产生肽的碱性框架(NH2-E(OtBu)-A-R(Pbf)-P-A-L-L-T(tBu)-S(tBu)-R(Pbf)L-R(Pbf)-F-I-P-K(Dde)-2-氯-三苯甲基树脂)。为选择性引入FITC至肽的C端,肽的N端经保护不受Boc影响。随后,将二碳酸二叔丁酯(30当量)与DIPEA(30当量)熔融在DMF中。添加DMF溶液至肽且在室温下培育达2小时,且肽以DMF、MeOH和DMF顺序洗涤。因此,获得Boc-E(OtBu)-A-R(Pbf)-P-A-L-L-T(tBu)-S(tBu)-R(Pbf)L-R(Pbf)-F-I-P-K(Dde)-2-氯-三苯甲基树脂。使用含2%肼的DMF来移除Dde以添加FITC至赖氨酸的C端,该Dde为C端残基赖氨酸的保护基。随后,将FITC(8当量)和DIPEA(16当量)熔融在DMF中,该DMF经添加至肽反应混合物,且混合物在室温下经培育达2小时,随后以DMF、MeOH、DMF顺序洗涤。因此,获得Boc-E(OtBu)-A-R(Pbf)-P-A-L-L-T(tBu)-S(tBu)-R(Pbf)L-R(Pbf)-F-I-P-K(FITC)-2-氯-三苯甲基树脂。TFA/TIS/H2O=95/2.5/2.5经添加以使肽与树脂分离。预冷却乙醚经添加至混合物,且使用离心分离作用来沉淀肽。在通过Prep-HPLC净化之后,纯度是以分析HPLC确定且分子重量系以LC/MS确定。所获得物质通过LC/MS证实分子重量而验证为pep1-FITC。随后缀合物经冷冻干燥。按照与上述pep1-FITC相同的方式,还制备了SEQ ID NO:1至SEQ ID NO:156的肽-FITC缀合物。
实施例3:pep(CCP)-FITC缀合物的细胞穿透性实验
(1)在HeLa细胞系中的细胞穿透性实验
细胞培养
HeLa细胞系人宫颈腺癌细胞系购自ATCC。将细胞在37℃下在5%CO2培育箱中在含有10%胎牛血清(Invitrogen,美国)、厄尔氏盐、非必要氨基酸、丙酮酸钠、100μg/mL青霉素及10单位/mL链霉素的最低必需培养基(Minimum Essential Medium;MEM)中培养。
流式细胞术和对细胞穿透的共聚焦显微镜分析
执行流式细胞术和共焦显微镜分析,以比较以SEQ ID NO:1至SEQ ID NO:156的肽、pep(CPP)和对照处理的细胞的细胞摄入的程度。
将细胞系在6孔培养板中分裂且在含有10%胎牛血清(Invitrogen公司,美国)、100μg/ml青霉素、100单位/ml链霉素的培养基中在37℃下在5%CO2培育箱中培养达12小时。在以PBS洗涤细胞系之后,在最低必需培养基中诱导饥饿达一小时。20uM的每一载体肽经处理且在37℃下培养达一小时。在重复三次以PBS洗涤细胞的步骤之后,将胰蛋白-EDTA在37℃下处理达10分钟以分离在细胞外部的载体肽。细胞以致冷的PBS收集且执行离心分离作用以重复洗涤细胞的步骤达三次。随后,细胞经悬浮在含有4%多聚甲醛的0.5ml PBS中且细胞的荧光使用FACS Calibur(美国BD公司)分析。比较对照物和与FITC结合的各种肽的细胞摄入态样且通过平均荧光强度(Mean Fluorescence Intensity;MFI)分析。
结果在图1至图23中显示。在图1至图23中所示的分析结果在下表3中详细给出。
[表3]
(2)在Huh 7细胞系中的细胞穿透性
细胞培养
Huh7(人肝细胞癌)细胞系系购自美国细胞培养收藏中心(American Type CellCulture;ATCC)并作为悬浮细胞使用。将该细胞在37℃下在5%CO2培育箱中在具有10%胎牛血清(Invitrogen,美国)、厄尔氏盐、非必要氨基酸、丙酮酸钠、100μg/ml青霉素及10单位/ml链霉素的MEM培养基中培养。
通过流式细胞术所用的细胞穿透性的筛选分析
为确认肽的细胞穿透性,以SEQ ID NO:1至SEQ ID NO:156处理Huh7细胞系,并通过流式细胞仪进行分析。还通过实施例(1)针对HeLa细胞所描述的方式来对所述分析方法进行确认。分析的结果在图24至图43中显示。
(3)在人T淋巴细胞系中的细胞穿透性实验
细胞培养
Jurkat细胞系(人T细胞白血病细胞系)系购自ATCC并作为悬浮细胞使用。且细胞在37℃下在5%CO2培育箱中在补充有10%胎牛血清(Invitrogen,美国)、厄尔氏盐、非必要氨基酸、丙酮酸钠、100μg/ml青霉素及100单位/ml链霉素的RPMI 1640培养基中培养。从健康人血液(50ml)中分离人来源的淋巴细胞,然后利用Biocoll分离溶液(Biochrom AG,柏林,德国)收集外周血单核细胞(PBMC)和淋巴细胞的层。
利用流式细胞术对细胞穿透性进行筛选分析
为确认肽的细胞穿透性,以SEQ ID NO:1至SEQ ID NO:156处理人T细胞淋巴细胞系,并通过流式细胞仪分析。还通过实施例(1)针对HeLa细胞所描述的方式来对所述分析方法进行确认。分析的结果在图52至图69中显示。
(4)细胞活性及细胞毒性的分析
将细胞系分在96孔培养板中,并在37℃下在5%CO2培育箱中在含有10%胎牛血清(Invitrogen,美国)、100μg/ml青霉素及10单位/ml链霉素的培养基中培养12小时。在以PBS洗涤细胞后,在最低必需培养基中诱导饥饿。20μM的各载体肽在37℃处理并培育1小时。在细胞培养后,通过MTT测定分析细胞活性及细胞毒性。结果在图59至图72中显示。
Claims (16)
1.一种细胞穿透载体肽在制备组合物中的应用,所述组合物包含细胞穿透载体肽与活性成分的缀合物,
其中,所述细胞穿透载体肽是人端粒酶逆转录酶的片段,
其中,所述细胞穿透载体肽具有20个氨基酸,并且由SEQ ID NO:48的氨基酸序列构成,
其中所述细胞穿透载体肽将所述活性成分输送至细胞内。
2.如权利要求1所述的应用,其中,所述活性成分为选自蛋白质、核酸、肽、脂质、二醇-脂质、矿物质、糖、纳米粒子、生物制品、对比造影剂和药物中的至少一种。
3.如权利要求1所述的应用,其中,所述活性成分为化合物。
4.如权利要求1所述的应用,其中,所述载体肽通过共价键与所述活性成分直接缀合或通过共价键与所述活性成分经接头间接缀合。
5.如权利要求1所述的应用,其中,所述载体肽及所述活性成分通过非共价键缀合。
6.如权利要求2所述的应用,其中,所述活性成分为蛋白质或肽。
7.如权利要求2所述的应用,其中,所述活性成分为细胞因子、抗体、抗体片段、治疗酶、可溶受体或配体。
8.如权利要求2所述的应用,其中,所述对比造影剂选自由不透射线对比造影剂、顺磁对比造影剂、超顺磁对比造影剂及CT对比造影剂组成的组。
9.如权利要求2所述的应用,其中,所述对比造影剂基于铁。
10.如权利要求9所述的应用,其中,所述对比造影剂为二茂铁羧酸盐。
11.如权利要求1至10中任一项所述的应用,其中,所述组合物是对比造影剂。
12.如权利要求11所述的应用,其中,所述对比造影剂用于对比造影细胞。
13.如权利要求12所述的应用,其中,所述细胞为干细胞。
14.如权利要求1至10中任一项所述的应用,其中,所述活性成分用于治疗或预防疾病,且所述组合物为药物组合物。
15.如权利要求1至10中任一项所述的应用,其中,所述活性成分为功能性化妆品的活性成分,且所述组合物为化妆品组合物。
16.如权利要求1至10中任一项所述的应用,其中,所述活性成分为健康功能食品的活性成分,且所述组合物为健康食品组合物。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910066919.3A CN110074997B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
CN201910066931.4A CN110092817B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
Applications Claiming Priority (11)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR20120104144 | 2012-09-19 | ||
KR10-2012-0104144 | 2012-09-19 | ||
KR20120104173 | 2012-09-19 | ||
KR10-2012-0104173 | 2012-09-19 | ||
KR20120109207 | 2012-09-28 | ||
KR10-2012-0109216 | 2012-09-28 | ||
KR20120109216 | 2012-09-28 | ||
KR10-2012-0109207 | 2012-09-28 | ||
KR20130017169 | 2013-02-18 | ||
KR10-2013-0017169 | 2013-02-18 | ||
PCT/KR2013/008438 WO2014046478A1 (ko) | 2012-09-19 | 2013-09-17 | 세포 투과성 펩티드, 그를 포함한 컨쥬게이트 및 그를 포함한 조성물 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910066919.3A Division CN110074997B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
CN201910066931.4A Division CN110092817B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104837859A CN104837859A (zh) | 2015-08-12 |
CN104837859B true CN104837859B (zh) | 2019-02-22 |
Family
ID=50341699
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910066919.3A Active CN110074997B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
CN201910066931.4A Active CN110092817B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
CN201380057740.2A Active CN104837859B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910066919.3A Active CN110074997B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
CN201910066931.4A Active CN110092817B (zh) | 2012-09-19 | 2013-09-17 | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 |
Country Status (8)
Country | Link |
---|---|
US (4) | US9902945B2 (zh) |
EP (3) | EP2899200B1 (zh) |
JP (5) | JP6352923B2 (zh) |
KR (3) | KR102544207B1 (zh) |
CN (3) | CN110074997B (zh) |
ES (2) | ES2956510T3 (zh) |
TW (3) | TWI655204B (zh) |
WO (1) | WO2014046478A1 (zh) |
Families Citing this family (33)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150014483A (ko) | 2012-05-11 | 2015-02-06 | 주식회사 카엘젬백스 | 항염증 활성을 갖는 펩티드 및 이를 포함하는 조성물 |
JP6267190B2 (ja) | 2012-05-11 | 2018-01-24 | ジェムバックス アンド カエル カンパニー,リミティド | 悪液質の予防用または治療用組成物 |
WO2014010971A1 (ko) | 2012-07-11 | 2014-01-16 | 주식회사 카엘젬백스 | 세포 투과성 펩티드, 그를 포함한 컨쥬게이트 및 그를 포함한 조성물 |
US20150125438A1 (en) | 2012-07-20 | 2015-05-07 | Sang Jae Kim | Anti-Inflammatory Peptides and Composition Comprising the Same |
WO2014046481A1 (ko) | 2012-09-19 | 2014-03-27 | 주식회사 카엘젬백스 | 세포 투과성 펩티드, 그를 포함한 컨쥬게이트 및 그를 포함한 조성물 |
EP2899200B1 (en) | 2012-09-19 | 2020-05-06 | Gemvax & Kael Co., Ltd. | Cell penetrating peptide, conjugate comprising same, and composition comprising conjugate |
TWI616530B (zh) * | 2012-09-19 | 2018-03-01 | 傑姆維克斯&凱爾有限公司 | 穿膜胜肽以及包含該胜肽之共軛物及組成物(一) |
JP6517692B2 (ja) | 2012-09-19 | 2019-05-22 | ジェムバックス アンド カエル カンパニー,リミティド | 細胞透過性ペプチド、それを含んだコンジュゲート、及びそれを含んだ組成物 |
US9907838B2 (en) | 2013-04-19 | 2018-03-06 | Gemvax & Kael Co., Ltd. | Composition and methods for treating ischemic damage |
AU2014275610B2 (en) | 2013-06-07 | 2018-06-14 | Gemvax & Kael Co., Ltd. | Biological markers useful in cancer immunotherapy |
EP3011967B1 (en) | 2013-06-21 | 2020-06-17 | Gemvax & Kael Co., Ltd. | Hormone secretion regulator, composition containing same, and method for controlling hormone secretion using same |
JP6382972B2 (ja) | 2013-10-23 | 2018-08-29 | ジェムバックス アンド カエル カンパニー,リミティド | 前立腺肥大治療用及びその予防用の組成物 |
EP3072519B1 (en) | 2013-11-22 | 2020-08-19 | Gemvax & Kael Co., Ltd. | Peptide having angiogenesis inhibitory activity and composition containing same |
ES2809251T3 (es) | 2013-12-17 | 2021-03-03 | Gemvax & Kael Co Ltd | Composición para tratar cáncer de próstata |
JP6420459B2 (ja) | 2014-04-11 | 2018-11-07 | ジェムバックス アンド カエル カンパニー,リミティド | 線維症抑制活性を有するペプチド及びこれを含む組成物 |
KR102232320B1 (ko) | 2014-04-30 | 2021-03-26 | 주식회사 젬백스앤카엘 | 장기, 조직 또는 세포 이식용 조성물, 키트 및 이식 방법 |
KR102413243B1 (ko) | 2014-12-23 | 2022-06-27 | 주식회사 젬백스앤카엘 | 안질환 치료 펩티드 및 이를 포함하는 안질환 치료용 조성물 |
JP6751097B2 (ja) | 2015-02-27 | 2020-09-02 | ジェムバックス アンド カエル カンパニー,リミティド | 聴力損傷予防用ペプチド及びそれを含む組成物 |
KR20180004752A (ko) | 2015-05-26 | 2018-01-12 | 주식회사 젬백스앤카엘 | 신규 펩티드 및 이를 포함한 조성물 |
KR101647804B1 (ko) * | 2015-06-24 | 2016-08-11 | 한국과학기술연구원 | 신규 세포투과 펩타이드 및 이의 용도 |
KR102638286B1 (ko) | 2015-07-02 | 2024-02-20 | 주식회사 젬백스앤카엘 | 항바이러스 활성 효능을 가지는 펩티드 및 이를 포함하는 조성물 |
EP3341389B1 (en) | 2015-08-28 | 2021-10-06 | The Medical College of Wisconsin, Inc. | Peptide inhibitors of telomerase translocation and therapeutic uses thereof |
ES2949697T3 (es) | 2016-04-07 | 2023-10-02 | Gemvax & Kael Co Ltd | Péptido que tiene efectos de aumento de la actividad telomerasa y de extensión de los telómeros, y composición que contienen el mismo |
KR101710186B1 (ko) * | 2016-09-01 | 2017-03-10 | (주)에이씨티 | 6-아미노헥사노산 유도체 및 그 유도체를 함유하는 광반응성 화장료 조성물 |
KR101843830B1 (ko) * | 2016-09-27 | 2018-04-02 | (주)라온크리에이트 | 피부 상태 개선 장치 및 그의 사용 방법 |
CN107168012A (zh) * | 2017-07-11 | 2017-09-15 | 深圳市华星光电技术有限公司 | 一种彩色光刻胶及其制备方法 |
JP7262105B2 (ja) | 2019-03-29 | 2023-04-21 | 国立研究開発法人理化学研究所 | 細胞透過性配列を有するポリペプチド及びそれを含む組成物 |
CN114213508B (zh) * | 2020-09-03 | 2024-06-04 | 深圳厚存纳米药业有限公司 | 一种多肽及其多肽复合物纳米粒、核酸疫苗和应用 |
KR20220142954A (ko) | 2021-04-15 | 2022-10-24 | 한국생명공학연구원 | 세포 투과 펩티드, 항암 펩티드 및 이를 포함하는 암 예방 또는 치료용 약학적 조성물 |
KR102415717B1 (ko) * | 2021-12-29 | 2022-07-05 | 주식회사 펩스젠 | 혈액뇌장벽 투과능을 가지는 신규한 펩타이드 및 이의 용도 |
WO2023128122A1 (ko) * | 2021-12-29 | 2023-07-06 | 주식회사 펩스젠 | 혈액뇌장벽 투과능을 가지는 펩타이드 및 이의 용도 |
KR102541402B1 (ko) * | 2022-02-18 | 2023-06-14 | 주식회사 레메디 | 아토피 피부염 예방 또는 치료 활성을 가지는 펩타이드 |
WO2024117321A1 (ko) * | 2022-12-01 | 2024-06-06 | ㈜에빅스젠 | 세포막 투과 펩티드 및 이를 포함하는 세포내 전달체 |
Family Cites Families (28)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6093809A (en) * | 1996-10-01 | 2000-07-25 | University Technology Corporation | Telomerase |
US6610839B1 (en) * | 1997-08-14 | 2003-08-26 | Geron Corporation | Promoter for telomerase reverse transcriptase |
CA2266752A1 (en) * | 1996-10-01 | 1998-04-09 | Geron Corporation | Telomerase reverse transcriptase |
US7585622B1 (en) * | 1996-10-01 | 2009-09-08 | Geron Corporation | Increasing the proliferative capacity of cells using telomerase reverse transcriptase |
US6475789B1 (en) * | 1996-10-01 | 2002-11-05 | University Technology Corporation | Human telomerase catalytic subunit: diagnostic and therapeutic methods |
US7030211B1 (en) * | 1998-07-08 | 2006-04-18 | Gemvax As | Antigenic peptides derived from telomerase |
US7078491B1 (en) | 2000-09-21 | 2006-07-18 | Amgen Inc. | Selective binding agents of telomerase |
US7294708B2 (en) * | 2002-05-31 | 2007-11-13 | Beijing Institute Of Biotechnology | Telomerase reverse transcriptase fragments and uses thereof |
WO2005107818A2 (en) | 2004-04-30 | 2005-11-17 | University Of Florida | Nanoparticles and their use for multifunctional bioimaging |
EP1994942A1 (en) * | 2007-05-25 | 2008-11-26 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Pharmaceutical compositions comprising telomerase, and uses thereof |
US8362209B2 (en) | 2007-08-23 | 2013-01-29 | University Of Medicine And Dentistry Of New Jersey | Telomerase reverse transcriptase variant |
AU2008304019A1 (en) * | 2007-09-25 | 2009-04-02 | Pastoral Greenhouse Gas Research Ltd | Phage Phi-mru polynucleotides and polypeptides and uses thereof |
WO2010003520A2 (en) * | 2008-06-16 | 2010-01-14 | Genovax S.R.L. | Anti-tumor immunotherapy |
US8252282B2 (en) * | 2008-06-19 | 2012-08-28 | University Of Medicine & Dentistry Of New Jersey | Nuclear telomerase reverse transcriptase variant |
IT1393509B1 (it) * | 2008-07-08 | 2012-04-27 | Advanced Accelerator Applications S A | Peptidi e loro uso come carrier in cellule cancerose |
EP2337795A2 (en) | 2008-10-01 | 2011-06-29 | Dako Denmark A/S | Mhc multimers in cancer vaccines and immune monitoring |
KR101169030B1 (ko) * | 2009-01-21 | 2012-07-26 | 애니젠 주식회사 | 신규한 세포막 투과 도메인 및 이를 포함하는 세포내 전달 시스템 |
JP6042211B2 (ja) * | 2010-02-16 | 2016-12-14 | ウルティモバックス エーエス | ポリペプチド |
KR20150014483A (ko) | 2012-05-11 | 2015-02-06 | 주식회사 카엘젬백스 | 항염증 활성을 갖는 펩티드 및 이를 포함하는 조성물 |
US20150125438A1 (en) | 2012-07-20 | 2015-05-07 | Sang Jae Kim | Anti-Inflammatory Peptides and Composition Comprising the Same |
US20170327802A1 (en) | 2012-07-20 | 2017-11-16 | Gemvax & Kael Co., Ltd. | Anti-Inflammatory Peptides and Composition Comprising the Same |
EP2899200B1 (en) | 2012-09-19 | 2020-05-06 | Gemvax & Kael Co., Ltd. | Cell penetrating peptide, conjugate comprising same, and composition comprising conjugate |
JP6517692B2 (ja) | 2012-09-19 | 2019-05-22 | ジェムバックス アンド カエル カンパニー,リミティド | 細胞透過性ペプチド、それを含んだコンジュゲート、及びそれを含んだ組成物 |
WO2014046481A1 (ko) | 2012-09-19 | 2014-03-27 | 주식회사 카엘젬백스 | 세포 투과성 펩티드, 그를 포함한 컨쥬게이트 및 그를 포함한 조성물 |
AU2014275610B2 (en) | 2013-06-07 | 2018-06-14 | Gemvax & Kael Co., Ltd. | Biological markers useful in cancer immunotherapy |
CN105531284B (zh) | 2013-07-12 | 2021-03-23 | 珍白斯凯尔有限公司 | 细胞穿透肽和包含其的缀合物 |
WO2017066349A1 (en) | 2015-10-13 | 2017-04-20 | Symic Ip, Llc | Ve-cadherin binding bioconjugate |
CN106749537A (zh) * | 2017-02-04 | 2017-05-31 | 北京大学第医院 | 一种多肽及其作为siRNA递送载体的应用 |
-
2013
- 2013-09-17 EP EP13838181.9A patent/EP2899200B1/en active Active
- 2013-09-17 TW TW107100939A patent/TWI655204B/zh active
- 2013-09-17 EP EP20172569.4A patent/EP3736282B1/en active Active
- 2013-09-17 TW TW107142413A patent/TWI678374B/zh active
- 2013-09-17 JP JP2015532963A patent/JP6352923B2/ja active Active
- 2013-09-17 KR KR1020227004140A patent/KR102544207B1/ko active IP Right Grant
- 2013-09-17 US US14/429,637 patent/US9902945B2/en active Active
- 2013-09-17 WO PCT/KR2013/008438 patent/WO2014046478A1/ko active Application Filing
- 2013-09-17 KR KR1020237019231A patent/KR102578890B1/ko active IP Right Grant
- 2013-09-17 CN CN201910066919.3A patent/CN110074997B/zh active Active
- 2013-09-17 CN CN201910066931.4A patent/CN110092817B/zh active Active
- 2013-09-17 KR KR1020157009509A patent/KR102362341B1/ko active IP Right Grant
- 2013-09-17 CN CN201380057740.2A patent/CN104837859B/zh active Active
- 2013-09-17 ES ES20172569T patent/ES2956510T3/es active Active
- 2013-09-17 EP EP23177458.9A patent/EP4230644A3/en active Pending
- 2013-09-17 TW TW102133731A patent/TWI655287B/zh active
- 2013-09-17 ES ES13838181T patent/ES2802253T3/es active Active
-
2018
- 2018-01-12 US US15/869,518 patent/US10822595B2/en active Active
- 2018-06-07 JP JP2018109473A patent/JP6697028B2/ja active Active
-
2020
- 2020-04-23 JP JP2020076850A patent/JP7041187B2/ja active Active
- 2020-09-25 US US17/032,781 patent/US11773381B2/en active Active
-
2022
- 2022-03-10 JP JP2022037094A patent/JP7368522B2/ja active Active
-
2023
- 2023-08-17 US US18/451,280 patent/US20240084273A1/en active Pending
- 2023-10-12 JP JP2023176669A patent/JP2023171606A/ja active Pending
Non-Patent Citations (2)
Title |
---|
Mitochondrial Medicine:Pharmacological targeting of mitochondria in disease;JS Armstrong;《British Journal of Pharmaclolgy》;20070521(第151期);第1157页图1d和第1157页右栏倒数第1段-1158左栏第1段 |
细胞穿膜肽作为药物载体的研究进展;任锦等;《药学学报》;20100112;第45卷(第1期);全文 |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104837859B (zh) | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 | |
CN104768967B (zh) | 细胞穿透性肽、包含该肽的缀合物、及包含该缀合物的组合物 | |
CN104981478B (zh) | 细胞穿透肽、包含该肽的缀合物、及包含该缀合物的组合物 | |
CN104822698B (zh) | 细胞穿透肽以及包含该肽的缀合物和组合物 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
EXSB | Decision made by sipo to initiate substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
CB02 | Change of applicant information |
Address after: South Korea field wide area Applicant after: Jane white Kell Co., Ltd. Applicant after: Jin Shangzai Address before: South Korea field wide area Applicant before: Jim Vickers & Kyle Ltd Applicant before: Jin Shangzai |
|
COR | Change of bibliographic data | ||
GR01 | Patent grant | ||
GR01 | Patent grant |