CN114656525B - RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs - Google Patents
RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs Download PDFInfo
- Publication number
- CN114656525B CN114656525B CN202210442337.2A CN202210442337A CN114656525B CN 114656525 B CN114656525 B CN 114656525B CN 202210442337 A CN202210442337 A CN 202210442337A CN 114656525 B CN114656525 B CN 114656525B
- Authority
- CN
- China
- Prior art keywords
- cancer
- cyclic peptide
- rgd cyclic
- peptide
- rgd
- 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
- 108010069514 Cyclic Peptides Proteins 0.000 title claims abstract description 89
- 102000001189 Cyclic Peptides Human genes 0.000 title claims abstract description 89
- 102000006495 integrins Human genes 0.000 title claims abstract description 57
- 108010044426 integrins Proteins 0.000 title claims abstract description 57
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 49
- 201000011510 cancer Diseases 0.000 title claims abstract description 31
- 239000003814 drug Substances 0.000 title claims abstract description 14
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 229940079593 drug Drugs 0.000 title abstract description 12
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 32
- 235000018417 cysteine Nutrition 0.000 claims abstract description 25
- 150000001413 amino acids Chemical group 0.000 claims abstract description 21
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000004471 Glycine Substances 0.000 claims abstract description 16
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000004473 Threonine Substances 0.000 claims abstract description 16
- 229960002898 threonine Drugs 0.000 claims abstract description 16
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims abstract description 12
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims abstract description 8
- FDKWRPBBCBCIGA-REOHCLBHSA-N (2r)-2-azaniumyl-3-$l^{1}-selanylpropanoate Chemical compound [Se]C[C@H](N)C(O)=O FDKWRPBBCBCIGA-REOHCLBHSA-N 0.000 claims abstract description 8
- 239000004475 Arginine Substances 0.000 claims abstract description 8
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 claims abstract description 8
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims abstract description 8
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims abstract description 8
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims abstract description 8
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims abstract description 8
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims abstract description 8
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims abstract description 8
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims abstract description 8
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims abstract description 8
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims abstract description 8
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims abstract description 8
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims abstract description 8
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims abstract description 8
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims abstract description 8
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims abstract description 8
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims abstract description 8
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims abstract description 8
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims abstract description 8
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims abstract description 8
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000004472 Lysine Substances 0.000 claims abstract description 8
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims abstract description 8
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims abstract description 8
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims abstract description 8
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960003767 alanine Drugs 0.000 claims abstract description 8
- 235000004279 alanine Nutrition 0.000 claims abstract description 8
- 229960003121 arginine Drugs 0.000 claims abstract description 8
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960001230 asparagine Drugs 0.000 claims abstract description 8
- 235000009582 asparagine Nutrition 0.000 claims abstract description 8
- 229960005261 aspartic acid Drugs 0.000 claims abstract description 8
- 235000003704 aspartic acid Nutrition 0.000 claims abstract description 8
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 235000013922 glutamic acid Nutrition 0.000 claims abstract description 8
- 239000004220 glutamic acid Substances 0.000 claims abstract description 8
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960002885 histidine Drugs 0.000 claims abstract description 8
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960000310 isoleucine Drugs 0.000 claims abstract description 8
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960003136 leucine Drugs 0.000 claims abstract description 8
- 229930182817 methionine Natural products 0.000 claims abstract description 8
- 229960005190 phenylalanine Drugs 0.000 claims abstract description 8
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960002429 proline Drugs 0.000 claims abstract description 8
- 229960001153 serine Drugs 0.000 claims abstract description 8
- 229960004441 tyrosine Drugs 0.000 claims abstract description 8
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960004295 valine Drugs 0.000 claims abstract description 8
- 239000004474 valine Substances 0.000 claims abstract description 8
- FDKWRPBBCBCIGA-UWTATZPHSA-N D-Selenocysteine Natural products [Se]C[C@@H](N)C(O)=O FDKWRPBBCBCIGA-UWTATZPHSA-N 0.000 claims abstract description 6
- ZFOMKMMPBOQKMC-KXUCPTDWSA-N L-pyrrolysine Chemical compound C[C@@H]1CC=N[C@H]1C(=O)NCCCC[C@H]([NH3+])C([O-])=O ZFOMKMMPBOQKMC-KXUCPTDWSA-N 0.000 claims abstract description 6
- ZKZBPNGNEQAJSX-UHFFFAOYSA-N selenocysteine Natural products [SeH]CC(N)C(O)=O ZKZBPNGNEQAJSX-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229940055619 selenocysteine Drugs 0.000 claims abstract description 6
- 235000016491 selenocysteine Nutrition 0.000 claims abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 44
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 14
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 7
- 208000009956 adenocarcinoma Diseases 0.000 claims description 7
- 229910052757 nitrogen Inorganic materials 0.000 claims description 7
- 201000000849 skin cancer Diseases 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 101710095468 Cyclase Proteins 0.000 claims description 6
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- GHITVUOBZBZMND-UHFFFAOYSA-N 1,3,5-tris(bromomethyl)benzene Chemical group BrCC1=CC(CBr)=CC(CBr)=C1 GHITVUOBZBZMND-UHFFFAOYSA-N 0.000 claims description 5
- 206010009944 Colon cancer Diseases 0.000 claims description 5
- 208000029742 colonic neoplasm Diseases 0.000 claims description 5
- 102000005962 receptors Human genes 0.000 claims description 5
- 108020003175 receptors Proteins 0.000 claims description 5
- 206010005003 Bladder cancer Diseases 0.000 claims description 4
- 208000003174 Brain Neoplasms Diseases 0.000 claims description 4
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 4
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 4
- 208000015634 Rectal Neoplasms Diseases 0.000 claims description 4
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 4
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 4
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 4
- 208000002495 Uterine Neoplasms Diseases 0.000 claims description 4
- 201000011199 bladder lymphoma Diseases 0.000 claims description 4
- 201000010881 cervical cancer Diseases 0.000 claims description 4
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 4
- 206010017758 gastric cancer Diseases 0.000 claims description 4
- 201000007270 liver cancer Diseases 0.000 claims description 4
- 208000014018 liver neoplasm Diseases 0.000 claims description 4
- 201000005202 lung cancer Diseases 0.000 claims description 4
- 208000020816 lung neoplasm Diseases 0.000 claims description 4
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 4
- 201000001441 melanoma Diseases 0.000 claims description 4
- 201000002528 pancreatic cancer Diseases 0.000 claims description 4
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 4
- 206010038038 rectal cancer Diseases 0.000 claims description 4
- 201000001275 rectum cancer Diseases 0.000 claims description 4
- 201000011549 stomach cancer Diseases 0.000 claims description 4
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 4
- 206010046766 uterine cancer Diseases 0.000 claims description 4
- 208000024770 Thyroid neoplasm Diseases 0.000 claims description 3
- 239000003112 inhibitor Substances 0.000 claims description 3
- 201000002510 thyroid cancer Diseases 0.000 claims description 3
- XBAXJCUNAGGGLR-JEDNCBNOSA-N (2s)-2,6-diaminohexanoic acid;1h-pyrrole Chemical compound C=1C=CNC=1.NCCCC[C@H](N)C(O)=O XBAXJCUNAGGGLR-JEDNCBNOSA-N 0.000 claims description 2
- 229960002449 glycine Drugs 0.000 claims 5
- 229960002989 glutamic acid Drugs 0.000 claims 2
- 229960002743 glutamine Drugs 0.000 claims 2
- 229960003646 lysine Drugs 0.000 claims 2
- 229960004452 methionine Drugs 0.000 claims 2
- 229960004799 tryptophan Drugs 0.000 claims 2
- 230000027455 binding Effects 0.000 abstract description 18
- 206010027476 Metastases Diseases 0.000 abstract description 4
- 230000009401 metastasis Effects 0.000 abstract description 4
- 230000035755 proliferation Effects 0.000 abstract description 3
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 48
- 210000004027 cell Anatomy 0.000 description 48
- 238000006243 chemical reaction Methods 0.000 description 23
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 18
- 238000010828 elution Methods 0.000 description 18
- 239000000243 solution Substances 0.000 description 17
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 15
- 238000000034 method Methods 0.000 description 14
- AMLYAMJWYAIXIA-VWNVYAMZSA-N cilengitide Chemical compound N1C(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](C(C)C)N(C)C(=O)[C@H]1CC1=CC=CC=C1 AMLYAMJWYAIXIA-VWNVYAMZSA-N 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 230000015572 biosynthetic process Effects 0.000 description 10
- 229950009003 cilengitide Drugs 0.000 description 10
- 102000004196 processed proteins & peptides Human genes 0.000 description 10
- 239000011347 resin Substances 0.000 description 10
- 229920005989 resin Polymers 0.000 description 10
- 238000003786 synthesis reaction Methods 0.000 description 10
- 108090000250 sortase A Proteins 0.000 description 9
- 230000005764 inhibitory process Effects 0.000 description 8
- 229920001184 polypeptide Polymers 0.000 description 8
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 7
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 238000004128 high performance liquid chromatography Methods 0.000 description 7
- 238000007363 ring formation reaction Methods 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 230000009471 action Effects 0.000 description 6
- 150000001945 cysteines Chemical class 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 102000009123 Fibrin Human genes 0.000 description 5
- 108010073385 Fibrin Proteins 0.000 description 5
- 108010047852 Integrin alphaVbeta3 Proteins 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000012071 phase Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 4
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 238000006555 catalytic reaction Methods 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 210000002744 extracellular matrix Anatomy 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 125000003367 polycyclic group Chemical group 0.000 description 4
- 125000006239 protecting group Chemical group 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 102000016359 Fibronectins Human genes 0.000 description 3
- 108010067306 Fibronectins Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical group CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- 239000013067 intermediate product Substances 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 238000010587 phase diagram Methods 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 2
- -1 Hexafluorophosphate Chemical compound 0.000 description 2
- 102100022337 Integrin alpha-V Human genes 0.000 description 2
- 108010040765 Integrin alphaV Proteins 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 239000003875 Wang resin Substances 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000033115 angiogenesis Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 238000009833 condensation Methods 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 229950003499 fibrin Drugs 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 229910052740 iodine Inorganic materials 0.000 description 2
- 239000011630 iodine Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000011535 reaction buffer Substances 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000010532 solid phase synthesis reaction Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 125000003396 thiol group Chemical group [H]S* 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 210000003556 vascular endothelial cell Anatomy 0.000 description 2
- LOQBLYVTXQGAEG-UHFFFAOYSA-N 1,3,5-tribromo-5-methylcyclohexa-1,3-diene Chemical compound BrC1(C)CC(=CC(=C1)Br)Br LOQBLYVTXQGAEG-UHFFFAOYSA-N 0.000 description 1
- 206010004433 Benign ovarian tumour Diseases 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 102000000844 Cell Surface Receptors Human genes 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 108090000279 Peptidyltransferases Proteins 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- NERFNHBZJXXFGY-UHFFFAOYSA-N [4-[(4-methylphenyl)methoxy]phenyl]methanol Chemical compound C1=CC(C)=CC=C1COC1=CC=C(CO)C=C1 NERFNHBZJXXFGY-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000004820 blood count Methods 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000009137 competitive binding Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 108091036078 conserved sequence Proteins 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 description 1
- 108010045325 cyclic arginine-glycine-aspartic acid peptide Proteins 0.000 description 1
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229940125798 integrin inhibitor Drugs 0.000 description 1
- 238000006192 iodination reaction Methods 0.000 description 1
- JDNTWHVOXJZDSN-UHFFFAOYSA-N iodoacetic acid Chemical compound OC(=O)CI JDNTWHVOXJZDSN-UHFFFAOYSA-N 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- CMWYAOXYQATXSI-UHFFFAOYSA-N n,n-dimethylformamide;piperidine Chemical compound CN(C)C=O.C1CCNCC1 CMWYAOXYQATXSI-UHFFFAOYSA-N 0.000 description 1
- 238000005580 one pot reaction Methods 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 201000008016 ovarian benign neoplasm Diseases 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000002221 trityl group Chemical group [H]C1=C([H])C([H])=C([H])C([H])=C1C([*])(C1=C(C(=C(C(=C1[H])[H])[H])[H])[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention belongs to the technical field of biology, and particularly relates to RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs. The RGD cyclic peptide is rich in disulfide bonds formed by cysteine sulfhydryl groups, and the amino acid sequence of the RGD cyclic peptide is as follows: g n (C y RGDfV) m CLPXT, wherein n is a positive integer from 1 to 3, y is 1 or 2, m is 1 or 2, x is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine. RGD cyclic peptides are directed against alpha in cancer cells v β 3 And alpha v β 5 The two integrins have strong selective binding capability, and can better inhibit metastasis and proliferation of cancer cells through selective binding of RGD cyclic peptide and the integrins.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs.
Background
RGD is a short peptide containing arginine-glycine-aspartic acid (Arg-Gly-Asp) and is mainly found in extracellular matrix (ECM) and Fibronectin of blood, for example, fibrin (FB), fibronectin (FN), laminin (Vitronnectin, VN) and Collagen (Collagen), and contains RGD sequence. RGD is a conserved sequence of these proteins and integrin receptors on cell membranes (e.g., alpha v β 3 And alpha v β 5 ) Interactions occur that mediate the adhesion of cells to the extracellular matrix. The over-expression of related integrin receptors on the cell membranes of many cancer cells plays an important role in cancer cell metastasis, proliferation, signaling, etc. Among the various integrin molecules, alpha v β 3 And alpha v β 5 Is a specific marker of tumor neovascularization and is closely related to tumor-induced angiogenesis. Integrin alpha v β 3 And alpha v β 5 Through mediating the adhesion of vascular endothelial cells and tumor cells, participate in angiogenesis and tumor metastasis, play a vital role in tumor growth. When its function is inhibited, vascular endothelial cells undergo apoptosis, and tumor growth is inhibited or even regressed. Therefore, it is found that a drug blocking the surface-specific integrin of tumor cells has very important significance in tumor therapy. On the other hand, the expression type of integrin in various tumor tissues is not completely consistent, and is closely related to the cell origin of tumor tissues and the malignancy of tumors. For example integrin alpha v β 3 High expression in malignant melanoma and malignant ovarian tumor cells; but is not expressed or expressed in tumor cells such as colon cancer, pancreatic cancer, breast cancer or benign ovarian tumor, etc., and is expressed in alpha v β 5 Or other types of integrins are expressed predominantly, thus the discovery and development of high affinity, highly selective tumor targeting peptides targeting specific subtypes of integrins is of significant interest for tumor targeting diagnosis and for studying the mechanisms of specific subtypes of integrins in the course of tumorigenesis and progression.
RGD sequence polypeptide as universal recognition site for adhesion of extracellular matrix and integrin receptor, and is used for developing targeted tumor cell integrin inhibitionThe basis of the agent. The cyclopeptide molecule cilengitide cyclo (RGDfNMeVal) from merck is the most prominent, however this compound ends up failing in clinical trials in phase three, suggesting a highly expressed alpha in tumor cells v β 3 And alpha v β 5 Further research is needed for peptide drugs with integrins as targets. In addition, due to alpha v β 3 And alpha v β 5 The complex is very similar in structure and function, and the currently discovered cyclic RGD polypeptide molecule pair alpha with good biological activity v β 3 And alpha v β 5 The poor selectivity of these two integrins also suggests that development of polypeptide molecules selectively inhibit alpha v β 3 And alpha v β 5 The study of these two integrins remains a great challenge, and thus there is a strong need to develop integrin inhibitors that are both biologically active and selective and can be used in the treatment and diagnosis of related tumors.
Disclosure of Invention
The application provides RGD cyclic peptide for cancer cell integrin and application thereof in preparing tumor medicaments so as to solve the problem of alpha of the existing polypeptide v β 3 And alpha v β 5 The technical problem of poor selectivity of the two integrins.
In a first aspect, the present application provides an RGD cyclic peptide for cancer cell integrins, said RGD cyclic peptide being enriched in disulfide bonds formed by cysteine sulfhydryl groups, said RGD cyclic peptide having the amino acid sequence: g n (C y RGDfV) m CLPXT, wherein n is a positive integer from 1 to 3, y is 1 or 2, m is 1 or 2, x is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine.
Optionally, in the RGD cyclic peptide, the nitrogen end of glycine and the carbon end of threonine are connected to form a ring through peptide bonds.
Optionally, the amino acid sequence of the RGD cyclic peptide comprises: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT and GGCCRGDfVCRGDfVCLPXT.
Alternatively, in the RGD cyclic peptide, cysteine is bonded with cysteine, 1,3, 5-Tribromotoluene (TBMB) to form disulfide bond.
Alternatively, the cancer cell integrin comprises an integrin of any one cancer cell of adenocarcinoma, colon cancer, lung cancer, rectal cancer, uterine cancer, cervical cancer, skin cancer, stomach cancer, liver cancer, brain tumor, bladder cancer and lymphoma; wherein the adenocarcinoma comprises pancreatic cancer and/or thyroid cancer, and the skin cancer comprises melanoma.
Alternatively, the receptor for the cancer cell integrin comprises alpha v β 3 、α v β 5 、α 5 β 1 、α v β 1 、α v β 6 、α Πb β 3 、α 2 β 1 、α 3 β 1 And alpha 4 β 1 。
Alternatively, the synthetic component of the RGD cyclic peptide comprises a linear peptide having the amino acid sequence GGGCRGDfVCLPXTGGS, GGGCRGDfVCRGDfVCLPXTGGS and GGCCRGDfVCRGDfVCLPXTGGS.
Alternatively, the linear peptide cyclase synthesizes RGD cyclic peptides, and the cyclase is a Sortase A enzyme.
In a second aspect, the present application provides a use of an RGD cyclic peptide for cancer cell integrin, said use comprising a use in the preparation of a cancer cell integrin inhibitor, said RGD cyclic peptide being enriched in disulfide bonds formed by cysteine sulfhydryl groups, said RGD cyclic peptide having the amino acid sequence: g n (C y RGDfV) m CLPXT, wherein n is a positive integer from 1 to 3, y is 1 or 2, m is 1 or 2, x is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine.
Optionally, the amino acid sequence of the RGD cyclic peptide comprises: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT and GGCCRGDfVCRGDfVCLPXT.
Compared with the prior art, the technical scheme provided by the embodiment of the application has the following advantages:
the RGD cyclic peptide provided by the embodiment of the application is rich in disulfide bonds formed by cysteine sulfhydryl groups, and the amino acid sequence of the RGD cyclic peptide is as follows: g n (C y RGDfV) m CLPXT, wherein n is a positive integer of 1-3, y is 1 or 2, m is 1 or 2, RGD cyclic peptide is specific for alpha in cancer cells v β 3 And alpha v β 5 The two integrins have strong selective binding capability, and can better inhibit metastasis and proliferation of cancer cells through selective binding of RGD cyclic peptide and the integrins.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and together with the description, serve to explain the principles of the invention.
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it will be obvious to those skilled in the art that other drawings can be obtained from these drawings without inventive effort.
FIG. 1 is a schematic diagram and characterization of GGGCRGDfVCLPXT cyclopeptide provided in the examples of the present application;
FIG. 2 is a schematic diagram and characterization of GGGCRGDfVCRGDfVCLPXT cyclic peptide provided in the examples of the present application;
FIG. 3 is a schematic diagram and characterization of the GGCCRGDfVCRGDfVCLPXT cyclopeptide provided in the examples of the present application.
Detailed Description
For the purposes of making the objects, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions of the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is apparent that the described embodiments are some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by one of ordinary skill in the art without undue burden from the present disclosure, are within the scope of the present application based on the embodiments herein.
Throughout the specification, unless specifically indicated otherwise, the terms used herein should be understood as meaning as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification will control. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the scope of the present invention. For example, room temperature may refer to a temperature in the range of 10 to 35 ℃.
Unless otherwise specifically indicated, the various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or may be prepared by existing methods.
The technical scheme of the embodiment of the application aims to solve the technical problems, and the overall thought is as follows:
according to an exemplary embodiment of the present invention, there is provided an RGD cyclic peptide for cancer cell integrins, which is rich in disulfide bonds formed by cysteine sulfhydryl groups, the amino acid sequence of the RGD cyclic peptide being as follows: g n (C y RGDfV) m CLPXT, wherein n is a positive integer from 1 to 3, y is 1 or 2, m is 1 or 2, x is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine.
Specifically, a linear polypeptide with RGD can be synthesized on resin by adopting a solid-phase polypeptide synthesis technology, trifluoroacetic acid is used for removing a side chain protecting group of the linear polypeptide and then is incubated with a transpeptidase Sortase A (Srt A), so that the linear peptide is cut off in situ on the resin and cyclized to form an RGD cyclic peptide product, and cysteine sulfhydryl is oxidized to form disulfide bonds; finally purifying to obtain the disulfide bond RGD-rich polycyclic peptide pure product, wherein the specific sequence comprises the following steps: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT, GGCCRGDfVCRGDfVCLPXT, GGCCRGDfVCLPXT, GCRGDfVCRGDfVCLPXT, GCCRGDfVCCRGDfVCLPXT.
In some embodiments, the nitrogen end of glycine is linked to the carbon end of threonine in a peptide bond to form a loop in the RGD cyclic peptide.
In the embodiment of the application, the nitrogen end of glycine and the carbon end of threonine are connected into a ring through a peptide bond, disulfide bonds are formed through cysteine sulfhydryl groups, a more complex polycyclic structure is obtained, and the more rigid annular structure has the characteristics of enzymolysis resistance, thermal stability, strong selective binding capacity and the like.
In some embodiments, the amino acid sequence of the RGD cyclic peptide comprises: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT and GGCCRGDfVCRGDfVCLPXT.
Specifically, the amino acid sequence of the RGD cyclic peptide is specifically as follows in se.q ID NO:1-SEQ ID NO: 3.
| Sequence number | Amino acid sequence |
| SE.Q ID NO：1 | GGGCRGDfVCLPXT |
| SE.Q ID NO：2 | GGGCRGDfVCRGDfVCLPXT |
| SE.Q ID NO：3 | GGCCRGDfVCRGDfVCLPXT |
The 3 RGD cyclic peptides have space conformation and have multi-ring structure characteristics rich in disulfide bonds, so that the 3 RGD cyclic peptides have better selective binding target property, thus having selective property when being specifically bound with cancer cell integrin, enhancing the selective binding capacity with specific integrin subtype (for example, specific selective binding integrin alpha v β 3 )。
In some embodiments, in the RGD cyclic peptide, cysteine is bonded to cysteine, 1,3, 5-Tribromomethylbenzene (TBMB) to form a disulfide bond.
Through bonding cysteine with cysteine and 1,3, 5-tribromomethyl benzene (TBMB) to form disulfide bond, the purpose of forming a polycyclic structure is achieved, and the rigid polycyclic structure has the characteristics of enzymolysis resistance, thermal stability, strong selective binding capacity and the like, and can remarkably improve the specific binding capacity with a target receptor (integrin).
In some embodiments, the cancer cell integrin comprises an integrin of any one of adenocarcinoma, colon cancer, lung cancer, rectal cancer, uterine cancer, cervical cancer, skin cancer, stomach cancer, liver cancer, brain tumor, bladder cancer, and lymphoma; wherein the adenocarcinoma comprises pancreatic cancer and/or thyroid cancer, and the skin cancer comprises melanoma.
In some embodiments, the receptor for the cancer cell integrin comprises alpha v β 3 、α v β 5 、α 5 β 1 、α v β 1 、
α v β 6 、α Πb β 3 、α 2 β 1 、α 3 β 1 And alpha 4 β 1 。
In particular, the receptors for cancer cell integrins include alpha v β 3 、α v β 5 、α 5 β 1 、α v β 1 、α v β 6 、α Πb β 3 、
α 2 β 1 、α 3 β 1 And alpha 4 β 1 Can be combined with RGD cyclic peptide, but the inventionRGD cyclic peptides involved in the invention can specifically bind to integrin alpha v β 3 Integrin alpha v β 3 The binding capacity is superior to other integrins, so that the method has the advantage of selective binding, and the developed subsequent anti-tumor drugs can be specifically aimed at specific tumor types.
In some embodiments, the synthetic component of the RGD cyclic peptide comprises a linear peptide having the amino acid sequence GGGCRGDfVCLPXTGGS, GGGCRGDfVCRGDfVCLPXTGGS and GGCCRGDfVCRGDfVCLPXTGGS.
The reason for selecting linear peptide with amino acid sequence GGGCRGDfVCLPXTGGS, GGGCRGDfVCRGDfVCLPXTGGS and GGCCRGDfVCRGDfVCLPXTGGS to synthesize RGD cyclic peptide is that cyclase A recognition sequence is added into the sequence, so that cyclic peptide is easy to synthesize by chemical enzyme method, and is simple and easy to operate and has high yield. In addition, the linear peptide is easy to design a disulfide-rich RGD polycyclic peptide structure. Therefore, the linear peptide is easy to design RGD polycyclic peptide structure rich in disulfide bonds, and is simple and easy to synthesize.
In some embodiments, the linear peptide cyclase synthesizes RGD cyclic peptides, which cyclase is a Sortase a enzyme.
In order to realize the cyclization of linear peptide, the Sortase A enzyme is selected, and the method has the positive effects of simplicity, easiness, higher yield and environmental protection.
According to another exemplary embodiment of the present invention, there is provided a use of an RGD cyclic peptide for cancer cell integrin, which comprises a use in preparing an inhibitor of cancer cell integrin, the RGD cyclic peptide being rich in disulfide bonds formed by cysteine sulfhydryl groups, the amino acid sequence of the RGD cyclic peptide being as follows: g n (C y RGDfV) m CLPXT, wherein n is a positive integer from 1 to 3, y is 1 or 2, m is 1 or 2, x is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine.
In some embodiments, the amino acid sequence of the RGD cyclic peptide comprises: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT and GGCCRGDfVCRGDfVCLPXT.
The method of the present invention will be described in detail with reference to examples, comparative examples and experimental data.
Example 1
RGD linear peptide was synthesized on Wang resin: GGGCRGDfVCLPXTGGS
(1) Swelling of the resin: fmoc-Ser (otBu) -Wang resin (load 0.564 mmol/g) was weighed into a polypeptide solid phase synthesis tube, DMF was added to submerge the resin and swelled for 3h. The resin was washed 3 times with MeOH, dichloromethane (DCM) and N, N-Dimethylformamide (DMF) in this order and dried for use.
(2) Removal of fluorenylmethoxycarbonyl (Fmoc): the resin was added to 5mL of 20% piperidine DMF solution and the reaction was shaken for 15min and repeated once, and finally the resin was washed 3 times with DMF.
(3) Condensation of the remaining amino acids: the resin was transferred to a polypeptide synthesizer (CEM) to effect the condensation of the remaining amino acids. The condensing agent is O-benzotriazol-tetramethylurea Hexafluorophosphate (HBTU), and the organic base is N, N-Diisopropylethylamine (DIPEA).
(4) Removing side chain protecting groups: the resin was eluted with DCM, dried by suction with a water pump for 20min, 2mL of trifluoroacetic acid cutting solution was slowly added under ice bath conditions, the reaction was stirred at room temperature for 2h, the cutting solution was filtered off, and washed 5 times with 98% acetic acid, DCM (5% DIPEA), DCM, DMF each. And (5) pumping the washed resin for later use.
One-pot synthesis of GGGCRGDfVCLPXT cyclic peptide
The linear peptide GGGCRGDfVCLPXTGGS undergoes an enzymatic reaction with the Sortase a enzyme and cyclizes (Sortase a recognizes the LPXTGG sequence and cleaves from between the amino acids threonine and glycine, linking the N-terminus of glycine to the C-terminus of threonine to form a cyclic peptide, fig. 1A). A solution of the Sortase A enzyme was added to a substrate peptide solution of 0.5mM to give a reaction concentration of 5. Mu.M, and the reaction buffer was supplemented to 400. Mu.L. 40 mu L of reaction solution in 0min,1h,3h,6h and 8.5h are sequentially taken, reaction buffer solution is added in equal volume, HPLC analysis is carried out (HPLC analysis parameters: mobile phase is acetonitrile and water, detection wavelength is 220nm, flow rate is 0.5mL/min, C18 reversed phase chromatographic column is 20% -50% acetonitrile is eluted for 60 min), and newly generated elution peak is collected for ESI mass spectrum analysis. Purifying with C-18 semi-preparative chromatographic column, and lyophilizing.
Example 2
GnCRGDfVCRGDfVCLPXT (N-terminal G forms an amide bond with C-segment T, n=3, y=1, m=2) cyclopeptide synthesis
The examples differ from example 1 in that the disulfide bond formation of three cysteines in the GnCRGDfVCRGDfVCLPXT cyclic peptide is mediated by 1,3, 5-Tribromomethylbenzene (TBMB) synthesis, which reacts with TBMB to form a tricyclic peptide structure (fig. 2A). In 600. Mu.L of the reaction system, the linear peptide GGGCRGDfVCRGDfVCLPKTGGS was 0.5mM, the Srt A reaction concentration was 5. Mu.M, and the TBMB reaction concentration was 1mM. HPLC liquid phase monitoring analysis of linear peptides reacted with TBMB for 20min,1h,3h,5h and Sortase A enzyme mediated substrate peptide reactions for 0h,1h,3h,6h,21h,29h. The HPLC analysis conditions were: mobile phases were water and acetonitrile, both containing 0.1% tfa; the flow rate is 0.5mL/min; the detection wavelength is 220nm; acetonitrile was eluted from 20% to 60% of the initial concentration gradient for 70min with 10 μl of reaction solution per sample.
Example 3
GnCCRGDfVCRGDfVCLPXT (N-terminal G forms an amide bond with C-segment T, n=2, y=2, m=2) cyclopeptide synthesis
The difference between the embodiment and the embodiment 1 is that: the 4 cysteines in GGCCRGDfVCRGDfVCLPXT cyclic peptide are selected to form disulfide bonds, the 1 st and 3 rd cysteines are selected to form disulfide bonds, and the 2 nd and 4 th cysteines are selected to form disulfide bonds. In the case of linear peptide solid phase synthesis, the thiol groups of 1 st and 3 rd cysteines were deprotected by trityl (trt), and the thiol groups of 1 st and 3 rd cysteines were subjected to Acm protection (FIG. 3A). In 600. Mu.L of the reaction system, the linear peptide GGCCRGDfVCRGDfVCLPXTLPKTGGS was 0.5mM, and the Srt A reaction concentration was 5. Mu.M, 37 ℃ The lower reaction is carried out for 24 hours to cyclize and form a first pair of disulfide bonds (Cys) 3 -Cys 10 ) Then an equal volume of 10eq (relative to Acm) iodoacetic acid solution (in which elemental iodine was dissolved in acetic acid beforehand) was added and reacted at room temperature for 3h to form a second pair of disulfide bonds (Cys) 4 -Cys 16 ). And sampled at different time periods of the reaction,20 mu L of the reaction solution is taken each time, HPLC monitoring is carried out, and the elution peak is collected for mass spectrum identification. The HPLC analysis conditions were: mobile phases were water and acetonitrile, both containing 0.1% tfa; the flow rate is
0.5mL/min; the detection wavelength is 220nm; acetonitrile was eluted from 20% to 60% of the initial concentration gradient for 70min with 10 μl of reaction solution per sample.
Example 4
RGD cyclic peptide and integrins on cell membranes are subjected to competitive binding detection, and the detection method comprises the following steps:
(1) Cell resuscitation: the cell line containing the 3 RGD cyclic peptides prepared and expressed in examples 1-3 was frozen in liquid nitrogen, immediately placed in a 37℃water bath for rapid thawing, 5mL of medium (SKOV-3 cells using 1640 medium; HEK-293 and HT-29 cells using DMEM medium) was added to the thawed cell frozen stock, centrifuged at 1000rpm for 5min, the supernatant was removed, the collected cells were resuspended in 5mL of medium, and after sufficient mixing, added to 25cm 2 The cells were cultured in a cell incubator at 37℃and 5% CO 2.
(2) Cell passage: when the cell fusion rate reaches about 90%, the cells can be subjected to subculture. Pipette up the culture medium, add 5mL sterile PBS to rinse the cells once, 25cm 2 Adding 0.5mL pancreatin into culture flask, placing into cell incubator at 37deg.C for digestion for about 3min, observing digestion condition under microscope at any time, adding 5mL culture medium to stop digestion when cells begin to fall off from the wall of culture flask, blowing off non-falling cells, mixing, transferring into 15mL sterile centrifuge tube, centrifuging at 1000rpm for 5min, pouring out supernatant, adding fresh culture medium 5mL resuspended cells, mixing thoroughly, inoculating 75cm 2 In a flask, the cells were cultured in a cell incubator at 37℃with 5% CO 2.
(3) Protein ligand coated 96-well plates: the FB protein is dissolved in the coating buffer solution, 100 mu L of the FB protein is added into each hole, and the concentration is 5 mu g/mL; VN protein was also coated at 100. Mu.L per well with a final concentration of 1. Mu.g/mL. To prevent experimental errors due to evaporation, the 96-well plates were sealed with a sealing film, covered with a plate cover, and left at 4 ℃ overnight.
(4) Washing the plate: pouring the coating liquid after the coating time is over, and lightly buckling on the water absorbing paper for 2 times to remove the coating liquid as much as possible; then 200. Mu.L of PBST buffer was added to each well, the 96-well plate was gently swabbed, and the PBST buffer was decanted and repeated twice.
(5) Closing: 200. Mu.L of 1% BSA (PBST configuration) was added to each well and blocked at 37℃for 1h.
(6) Washing the plate: pouring the sealing liquid, lightly buckling on the absorbent paper for 2 times, and adding 200 mu L of PBST into each hole for washing once; then washed 2 times with PBS solution. The absorbent paper is buckled to be dry, and cells are inoculated for standby.
(7) Inoculating cells: the 96-well plate coated with FB protein is inoculated with integrin positive cell line human ovarian cancer cells SKOV-3 and human embryo kidney cells HEK-293, and the 96-well plate coated with VN protein is inoculated with HT-29 cells. Taking cells with good growth vigor, performing pancreatin digestion, performing centrifugal resuspension, fully and uniformly mixing, counting by using a blood cell counting plate, and finally inoculating 10000 cells per hole, wherein the inoculation volume is 25 mu L; meanwhile, the cyclic peptide drugs are prepared by a culture medium and diluted into different concentration gradients according to 10 times, three holes are repeated for each concentration, and the volume of the drugs which are expected to be added in each hole is 25 mu L; then mixing the cell sap with 25 μl of each drug to allow interaction of cell surface receptor protein and cyclopeptide drug for 30min, adding 96-well plate, and placing at 37deg.C CO 2 Culturing in an incubator for 2 hours. Cilengitide (Cilengitide) at different concentration gradients was used as a positive control, without any drug wells (other solvents were added) as a control, and without any drug and solvent as a blank.
(8) Thiazole blue (MTT) action: the syringe was carefully aspirated, 100. Mu.L of buffer (PBS) was added to each well, the 96-well plate was gently shaken to wash unbound cells, 20. Mu.L of MTT solution and 80. Mu.L of PBS were added to each well after the PBS was aspirated, and the 96-well plate was placed in an incubator at 37℃for 4 hours.
(9) And (3) light absorption value detection: after the MTT reaction was completed, the MTT solution was carefully removed, 100. Mu.L of DMSO was added, and the absorbance was measured at 490nm using a microplate reader after 10min at 37℃with shaking bed 120 rpm.
(10) And (3) data processing: cell adhesion inhibition ratio= (experimental group absorption value-control group absorption value)/control group absorption value. Statistical analysis was performed using the OriginPro 8 software (n=3) and the selection was calculated as the inhibition ratio of αvβ5 to αvβ3, data shown in table 1.
Table 1.
As can be seen from the table, the inhibition activity of cilengitide against integrin αvβ3 was 0.11, whereas the best inhibitory effect against αvβ3 in the 3 cyclic peptides of the present application was GGGCRGDfVCRGDfVCLPXT cyclic peptide, which had a value of 0.20, and the inhibition activity was close to that of the positive control cilengitide (0.20 vs 0.11). However, compared with the cilengitide, the GGGCRGDfVCRGDfVCLPXT cyclic peptide has better selective binding capacity, can specifically bind integrin alpha v beta 3, the selective value of the GGGCRGDfVCRGDfVCLPXT cyclic peptide reaches 151:1, and the selective binding capacity of the cilengitide class 3 cyclic peptide to alpha v beta 5/alpha v beta 3 is high. Wherein, in the detection, the types of X comprise glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenium cysteine and pyrrole lysine, and the verification is carried out one by one, and the average value is obtained.
In addition, it is clear from the table that the inhibition rate of adhesion of GGGCRGDfVCLPXT cyclic peptide to integrin αvβ3 is 1.54. Mu.M, the selective binding capacity of integrin αvβ5 to αvβ3 reaches 38.9:1, which is higher than that of cilengitide (3.1:1), indicating that the selective binding capacity of the cyclic peptide to integrin is better. The adhesion inhibition rate of GGGCRGDfVCRGDfVCLPXT cyclic peptide to integrin alpha v beta 3 is 0.20 mu M, the inhibition activity is close to that of positive control cilengitide (0.11 mu M), but the selective binding capacity of the integrin alpha v beta 5 to alpha v beta 3 is up to 151:1, which is far higher than that of cilengitide (3.1:1), the activity is also higher than that of other two cyclic peptides, which indicates that the tricyclic structure and disulfide bond-rich conformation in the cyclic peptide can promote RGD motif to selectively bind integrin, and the specific binding capacity is improved;
the inhibition rate of the GGCCRGDfVCRGDfVCLPXT cyclic peptide on the integrin alpha v beta 3 is 8.43 mu M, the selective binding capacity of the integrin alpha v beta 5 and alpha v beta 3 reaches 24.6:1, which is higher than that of the cilengitide (3.1:1), thus indicating that the selective binding capacity of the cyclic peptide on the integrin is better.
Detailed description of the drawings 1-3:
as shown in FIG. 1, FIG. 1A is a schematic representation of the synthesis of GGGCRGDfVCLPXT cyclic peptide by the linear peptide GGGCRGDfVCLPXTGGS under the catalysis of Sortase A. FIG. 1B is a liquid phase diagram of the synthesis process with elution times on the abscissa. The initial state of the Sortase A cyclization reaction is the initial state of the linear peptide GGGCRGDfVCLPXTGGS at 0min in the ordinate, wherein the elution time 26.787min is the elution peak of the linear peptide GGGCRGDfVCLPXTGGS. In the enzyme catalytic reaction for 15min, new elution peaks appear in the graphs for 33.058min and 35.079min respectively, and when the reaction is carried out for 1h and 9h, the sizes of elution peaks for 33.058min and 35.079min are converted, the elution peak for 35.079min is gradually reduced, and the elution peak for 33.058min is gradually increased, which means that the elution peak material for 35.079min is a cyclized intermediate product, which realizes head-to-tail cyclization, but disulfide bonds are not oxidized into bonds (specific mass spectrogram is shown in fig. 1C, molecular weight is 1391.64), the elution peak material for 33.058min is a final cyclized product GGGCRGDfVCLPXT, and the mass spectrogram of the cyclized product is shown in fig. 1D and is 1389.64.
As shown in FIG. 2, FIG. 2A is a schematic diagram of the synthesis of GGGCRGDfVCRGDfVCLPXTGGS as a linear peptide under Sortase A catalysis. FIG. 2B is a liquid phase diagram of the synthesis process, the abscissa shows the elution time, the ordinate shows the HPLC real-time monitoring graph of the linear peptide added with 1,3, 5-Tribromomethylbenzene (TBMB) for 25 minutes, 36.389min shows the product elution peak of the linear peptide GGGCRGDfVCRGDfVCLPXTGGS after the reaction of cysteine and TBMB, and 38.061min shows the mass spectrum data (see FIG. 2C). When the reaction proceeds for 5 hours, the linear peptide substrate is substantially converted to an intermediate product. And then adding Sortase A enzyme to realize end-to-end cyclization, adding enzyme to catalyze the reaction for 1h,6h and 29h, gradually increasing the peak area of an elution peak for 26.808min, and supposing that the substance of the peak is a cyclized final product GGGCRGDfVCRGDfVCLPXT, wherein the molecular weight of the peak is 2184.04 (figure 2D) through mass spectrum identification, and the molecular weight of the peak is consistent with that of a final cyclic peptide product GGGCRGDfVCRGDfVCLPXT, so that the elution peak is the final cyclized product.
As shown in fig. 3, fig. 3A is a schematic diagram of synthesis of GGCCRGDfVCRGDfVCLPXTGGS under the catalysis of Sortase a, fig. 3B is a liquid phase diagram of the synthesis process, the abscissa is the elution time, 0min in the ordinate is the initial state of the linear peptide enzymatic reaction, and the peak 27.671min is the linear peptide GGCCRGDfVCRGDfVCLPXTGGS. When the reaction proceeded to 15min, a new elution peak was seen for 32.55min, which was identified by mass spectrometry as a linear peptide end-to-end cyclization product (FIG. 3C), at which time neither pair of disulfide bonds was formed. As the reaction proceeds to 1h, oxygen in the air oxidizes the exposed cysteine side chain thiol to form disulfide bonds, a new elution peak appears for 30.24min, indicating the generation of a new product, which is identified by mass spectrometry as an intermediate cyclization product forming a pair of disulfide bonds (mass spectrometry see 3C), and all of the substrate is converted to such intermediate product as the reaction proceeds to 6 h. The other cysteine side chain sulfhydryl uses orthogonal protecting group Acm protection, and because the disulfide bond cannot be formed under the action of oxygen at the moment, the Acm protecting group is removed under the action of iodine simple substance, the exposed sulfhydryl forms disulfide bond, after iodination reaction for 3h, the eluting peak appears in 31.54min as the final cyclization product (mass spectrum is under 3C), is rich in two pairs of disulfide bonds, and is a selective bond.
It should be noted that in this document, relational terms such as "first" and "second" and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.
The foregoing is only a specific embodiment of the invention to enable those skilled in the art to understand or practice the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (7)
1. An RGD cyclic peptide for use in cancer cell integrins, wherein the RGD cyclic peptide is enriched in disulfide bonds formed by cysteine sulfhydryl groups, the amino acid sequence of the RGD cyclic peptide being: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT and GGCCRGDfVCRGDfVCLPXT;
in the RGD cyclic peptide, the nitrogen end of the first glycine and the carbon end of the last threonine are connected into a ring through peptide bonds; wherein X is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine and pyrrolysine.
2. The RGD cyclic peptide of claim 1, wherein in the RGD cyclic peptide, cysteine is bonded to cysteine, 1,3, 5-Tribromomethylbenzene (TBMB) to form a disulfide bond.
3. The RGD cyclic peptide according to claim 1, wherein the cancer cell integrin comprises an integrin of any one cancer cell of adenocarcinoma, colon cancer, lung cancer, rectal cancer, uterine cancer, cervical cancer, skin cancer, stomach cancer, liver cancer, brain tumor, bladder cancer and lymphoma; wherein the adenocarcinoma comprises pancreatic cancer and/or thyroid cancer, and the skin cancer comprises melanoma.
4. The RGD cyclic peptide of claim 1, wherein the receptor for the cancer cell integrin comprises a v β 3 。
5. The RGD cyclic peptide of claim 1, wherein the synthetic component of the RGD cyclic peptide comprises a linear peptide having the amino acid sequence GGGCRGDfVCLPXTGGS, GGGCRGDfVCRGDfVCLPXTGGS and ggccrgdcgfvcrgdclpxtggs; the nitrogen end of the first glycine and the carbon end of the last threonine in the RGD cyclic peptide are connected into a ring through peptide bonds.
6. The RGD cyclic peptide of claim 5, wherein the linear peptide is synthesized from RGD cyclic peptide by a cyclase, which is a Sortase a enzyme.
7. Use of an RGD cyclic peptide for cancer cell integrins for the preparation of a tumor medicament, the use comprising the use in the preparation of a cancer cell integrins inhibitor, the RGD cyclic peptide being enriched in disulfide bonds formed by cysteine sulfhydryl groups, the amino acid sequence of the RGD cyclic peptide being: GGGCRGDfVCLPXT, GGGCRGDfVCRGDfVCLPXT and GGCCRGDfVCRGDfVCLPXT; in the RGD cyclic peptide, the nitrogen end of the first glycine and the carbon end of the last threonine are connected into a ring through peptide bonds; x is any one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenium cysteine and pyrrole lysine; wherein the tumor is any one of adenocarcinoma, colon cancer, lung cancer, rectal cancer, uterine cancer, cervical cancer, skin cancer, gastric cancer, liver cancer, brain tumor, bladder cancer and lymphoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210442337.2A CN114656525B (en) | 2022-04-25 | 2022-04-25 | RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210442337.2A CN114656525B (en) | 2022-04-25 | 2022-04-25 | RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114656525A CN114656525A (en) | 2022-06-24 |
| CN114656525B true CN114656525B (en) | 2024-02-02 |
Family
ID=82037711
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210442337.2A Active CN114656525B (en) | 2022-04-25 | 2022-04-25 | RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114656525B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1002543A1 (en) * | 1998-11-12 | 2000-05-24 | Jcr Pharmaceuticals Co., Ltd. | Pharmaceutical compositions for the therapy of glioma or prostate cell proliferative disorders containing cyclic rgd compounds |
| CN101492494A (en) * | 2009-03-05 | 2009-07-29 | 浙江大学 | Integrin ligand cyclic peptide analogues and cyclization method |
| CN101638429A (en) * | 2009-08-28 | 2010-02-03 | 南方医科大学 | Small molecular peptide of specificity recognition cell surface integrin alphal3beta1 |
| CN108138204A (en) * | 2015-09-25 | 2018-06-08 | 豪夫迈·罗氏有限公司 | Use the method for sorting enzyme A production thioesters |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030125243A1 (en) * | 2000-07-20 | 2003-07-03 | Jun Liu | Synthesis of cyclic peptides |
| US10532106B2 (en) * | 2014-10-29 | 2020-01-14 | Bicyclerd Limited | Bicyclic peptide ligands specific for MT1-MMP |
| GB201900528D0 (en) * | 2019-01-15 | 2019-03-06 | Bicyclerd Ltd | Bicyclic peptide ligands specific for integrin AVB3 |
-
2022
- 2022-04-25 CN CN202210442337.2A patent/CN114656525B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1002543A1 (en) * | 1998-11-12 | 2000-05-24 | Jcr Pharmaceuticals Co., Ltd. | Pharmaceutical compositions for the therapy of glioma or prostate cell proliferative disorders containing cyclic rgd compounds |
| CN101492494A (en) * | 2009-03-05 | 2009-07-29 | 浙江大学 | Integrin ligand cyclic peptide analogues and cyclization method |
| CN101638429A (en) * | 2009-08-28 | 2010-02-03 | 南方医科大学 | Small molecular peptide of specificity recognition cell surface integrin alphal3beta1 |
| CN108138204A (en) * | 2015-09-25 | 2018-06-08 | 豪夫迈·罗氏有限公司 | Use the method for sorting enzyme A production thioesters |
Non-Patent Citations (3)
| Title |
|---|
| Chen Y,等.Sortase A-mediated cyclization of novel polycyclic RGD peptides for ανβ3 integrin targeting.《Bioorg Med Chem Lett.》.2022,全文. * |
| New potent and selective avb3 integrin ligands: Macrocyclic peptides containing RGD motif synthesized by sortase A-mediated ligation;Zhimeng Wu等;《Bioorganic & Medicinal Chemistry Letters》;第27卷;摘要，正文第1912页左栏第1段，图1，方案1，表2 * |
| 靶向整合素α_vβ_3放射性核素标记分子探针优化策略的研究进展;陈皓;吴华;;核技术(第09期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114656525A (en) | 2022-06-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Luo et al. | Designer D-form self-assembling peptide nanofiber scaffolds for 3-dimensional cell cultures | |
| WO2012121057A1 (en) | Novel cross-linked peptides containing non-peptide cross-linked structure, method for synthesizing cross-linked peptides, and novel organic compound used in method | |
| CN113292635B (en) | Polypeptide targeting CD47 and application thereof | |
| US10526370B2 (en) | Pentapeptide associated with integrin receptor alpha vbeta3 | |
| Taguchi et al. | Disulfide-driven cyclic peptide synthesis of human endothelin-2 with a solid-supported Npys-Cl | |
| CN111040020B (en) | Alkene thioether staple peptide and preparation method and application thereof | |
| Bucci et al. | On-resin multicomponent 1, 3-dipolar cycloaddition of cyclopentanone–proline enamines and sulfonylazides as an efficient tool for the synthesis of amidino depsipeptide mimics | |
| CN114656525B (en) | RGD cyclic peptide for cancer cell integrin and application thereof in preparation of tumor drugs | |
| Bazureau et al. | Ultrasound and microwave: recent advances in organic chemistry | |
| De Luca et al. | A Late-Stage synthetic approach to lanthionine-containing peptides via S-alkylation on cyclic sulfamidates promoted by molecular sieves | |
| CN112480211B (en) | Anti-tumor polypeptide targeting PKM2 protein and application thereof | |
| CN106749524B (en) | Anti-obesity heptapeptide NPVWKRK | |
| CN107446022B (en) | Polypeptide PIP-14 capable of antagonizing RNA binding activity of PARP1 protein and application thereof | |
| Todorovic et al. | Rationally designed amanitins achieve enhanced cytotoxicity | |
| CN109248324B (en) | GnRH analogue-anti-tumor drug conjugate, preparation method and application thereof | |
| JP4657721B2 (en) | Synthesis and characterization of novel guidance systems and vectorization of therapeutic molecules into target cells | |
| CN106699850A (en) | RBBP4 targeting polypeptide and anti-tumor polypeptide, and applications thereof | |
| CN114276411B (en) | Solid phase synthesis method of cyclic peptide compound containing trithioether bond | |
| CN112358531B (en) | Polypeptide targeting HER2 protein and application thereof | |
| CN111518171B (en) | Polypeptide targeting human hepatoma cells and application thereof | |
| CN110272471A (en) | A kind of preparation method of tumour medicine made of polypeptide, polypeptide and tumour medicine | |
| CN112830975B (en) | Alpha-helical conformation stable apoptosis-promoting bicyclic polypeptide and preparation method and application thereof | |
| CN117186186A (en) | Novel cyclic peptide compound and synthetic method and application thereof | |
| CN112390859B (en) | Self-assembly polypeptide probe for identifying Caspase protein, preparation method and application | |
| CN116947969A (en) | PD-L1 targeting polypeptide, preparation method, assembled nanosphere and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |