CN116747317A - 靶向b7-h3抗体-mmae偶联物及其应用 - Google Patents
靶向b7-h3抗体-mmae偶联物及其应用 Download PDFInfo
- Publication number
- CN116747317A CN116747317A CN202310475213.9A CN202310475213A CN116747317A CN 116747317 A CN116747317 A CN 116747317A CN 202310475213 A CN202310475213 A CN 202310475213A CN 116747317 A CN116747317 A CN 116747317A
- Authority
- CN
- China
- Prior art keywords
- antibody
- solution
- compound
- reaction
- drug
- 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.)
- Pending
Links
- 230000008685 targeting Effects 0.000 title description 11
- 239000000611 antibody drug conjugate Substances 0.000 claims abstract description 60
- 229940049595 antibody-drug conjugate Drugs 0.000 claims abstract description 60
- 150000001875 compounds Chemical class 0.000 claims abstract description 25
- 201000010099 disease Diseases 0.000 claims abstract description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 12
- 239000000243 solution Substances 0.000 claims description 46
- 238000006243 chemical reaction Methods 0.000 claims description 25
- 206010028980 Neoplasm Diseases 0.000 claims description 22
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 claims description 21
- 239000003814 drug Substances 0.000 claims description 20
- 229940079593 drug Drugs 0.000 claims description 18
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 14
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 14
- 239000002904 solvent Substances 0.000 claims description 14
- 238000002360 preparation method Methods 0.000 claims description 12
- 229940125782 compound 2 Drugs 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 230000002062 proliferating effect Effects 0.000 claims description 9
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 claims description 8
- IEDXPSOJFSVCKU-HOKPPMCLSA-N [4-[[(2S)-5-(carbamoylamino)-2-[[(2S)-2-[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-3-[[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate Chemical compound CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)c1ccccc1)OC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCc1ccc(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN2C(=O)CCC2=O)C(C)C)cc1)C(C)C IEDXPSOJFSVCKU-HOKPPMCLSA-N 0.000 claims description 8
- 108010093470 monomethyl auristatin E Proteins 0.000 claims description 8
- 230000035484 reaction time Effects 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 7
- 201000011510 cancer Diseases 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 206010018338 Glioma Diseases 0.000 claims description 5
- 239000000427 antigen Substances 0.000 claims description 5
- 102000036639 antigens Human genes 0.000 claims description 5
- 108091007433 antigens Proteins 0.000 claims description 5
- 229940125904 compound 1 Drugs 0.000 claims description 5
- 229940126214 compound 3 Drugs 0.000 claims description 5
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 5
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 claims description 5
- 235000010378 sodium ascorbate Nutrition 0.000 claims description 5
- 229960005055 sodium ascorbate Drugs 0.000 claims description 5
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 claims description 5
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 claims description 4
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 4
- 208000032612 Glial tumor Diseases 0.000 claims description 4
- 125000005647 linker group Chemical group 0.000 claims description 4
- YUOCYTRGANSSRY-UHFFFAOYSA-N pyrrolo[2,3-i][1,2]benzodiazepine Chemical compound C1=CN=NC2=C3C=CN=C3C=CC2=C1 YUOCYTRGANSSRY-UHFFFAOYSA-N 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 238000002626 targeted therapy Methods 0.000 claims description 4
- MFRNYXJJRJQHNW-DEMKXPNLSA-N (2s)-2-[[(2r,3r)-3-methoxy-3-[(2s)-1-[(3r,4s,5s)-3-methoxy-5-methyl-4-[methyl-[(2s)-3-methyl-2-[[(2s)-3-methyl-2-(methylamino)butanoyl]amino]butanoyl]amino]heptanoyl]pyrrolidin-2-yl]-2-methylpropanoyl]amino]-3-phenylpropanoic acid Chemical compound CN[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MFRNYXJJRJQHNW-DEMKXPNLSA-N 0.000 claims description 3
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 3
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 108010059074 monomethylauristatin F Proteins 0.000 claims description 3
- LGZKGOGODCLQHG-CYBMUJFWSA-N 5-[(2r)-2-hydroxy-2-(3,4,5-trimethoxyphenyl)ethyl]-2-methoxyphenol Chemical compound C1=C(O)C(OC)=CC=C1C[C@@H](O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-CYBMUJFWSA-N 0.000 claims description 2
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 claims description 2
- 229930126263 Maytansine Natural products 0.000 claims description 2
- 150000001540 azides Chemical class 0.000 claims description 2
- HXCHCVDVKSCDHU-LULTVBGHSA-N calicheamicin Chemical compound C1[C@H](OC)[C@@H](NCC)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@](C/3=C/CSSSC)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HXCHCVDVKSCDHU-LULTVBGHSA-N 0.000 claims description 2
- 229930195731 calicheamicin Natural products 0.000 claims description 2
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical class C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 claims description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 2
- LGZKGOGODCLQHG-UHFFFAOYSA-N combretastatin Natural products C1=C(O)C(OC)=CC=C1CC(O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-UHFFFAOYSA-N 0.000 claims description 2
- 229960000684 cytarabine Drugs 0.000 claims description 2
- 229960004679 doxorubicin Drugs 0.000 claims description 2
- 229960005277 gemcitabine Drugs 0.000 claims description 2
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 claims description 2
- WKPWGQKGSOKKOO-RSFHAFMBSA-N maytansine Chemical compound CO[C@@H]([C@@]1(O)C[C@](OC(=O)N1)([C@H]([C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(C)=O)CC(=O)N1C)C)[H])\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 WKPWGQKGSOKKOO-RSFHAFMBSA-N 0.000 claims description 2
- ANZJBCHSOXCCRQ-FKUXLPTCSA-N mertansine Chemical compound CO[C@@H]([C@@]1(O)C[C@H](OC(=O)N1)[C@@H](C)[C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(=O)CCS)CC(=O)N1C)\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 ANZJBCHSOXCCRQ-FKUXLPTCSA-N 0.000 claims description 2
- JFCFGYGEYRIEBE-YVLHJLIDSA-N wob38vs2ni Chemical compound CO[C@@H]([C@@]1(O)C[C@H](OC(=O)N1)[C@@H](C)[C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(=O)CCC(C)(C)S)CC(=O)N1C)\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 JFCFGYGEYRIEBE-YVLHJLIDSA-N 0.000 claims description 2
- 150000001345 alkine derivatives Chemical class 0.000 claims 1
- 239000004007 alpha amanitin Substances 0.000 claims 1
- 150000001412 amines Chemical class 0.000 claims 1
- 239000012153 distilled water Substances 0.000 claims 1
- VQNATVDKACXKTF-XELLLNAOSA-N duocarmycin Chemical compound COC1=C(OC)C(OC)=C2NC(C(=O)N3C4=CC(=O)C5=C([C@@]64C[C@@H]6C3)C=C(N5)C(=O)OC)=CC2=C1 VQNATVDKACXKTF-XELLLNAOSA-N 0.000 claims 1
- 229960005501 duocarmycin Drugs 0.000 claims 1
- 229930184221 duocarmycin Natural products 0.000 claims 1
- 239000008055 phosphate buffer solution Substances 0.000 claims 1
- 150000003839 salts Chemical class 0.000 abstract description 15
- 201000007983 brain glioma Diseases 0.000 abstract description 8
- 239000000562 conjugate Substances 0.000 abstract description 7
- 230000003463 hyperproliferative effect Effects 0.000 abstract description 3
- 230000003902 lesion Effects 0.000 abstract description 3
- 229960000074 biopharmaceutical Drugs 0.000 abstract description 2
- 239000012453 solvate Substances 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 54
- 241000699670 Mus sp. Species 0.000 description 12
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 10
- 239000000872 buffer Substances 0.000 description 9
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 9
- 239000002953 phosphate buffered saline Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- -1 ammonium cations Chemical class 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 241000282412 Homo Species 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 238000011033 desalting Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 3
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 3
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 230000005526 G1 to G0 transition Effects 0.000 description 3
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 108010019160 Pancreatin Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 150000001450 anions Chemical class 0.000 description 3
- 150000001768 cations Chemical class 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 3
- 229940055695 pancreatin Drugs 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- DOMDXTIMIZCSNC-UHFFFAOYSA-N (2Z)-2-[(2E,4E)-5-[3-[6-(2,5-dioxopyrrolidin-1-yl)oxy-6-oxohexyl]-1,1-dimethyl-6,8-disulfobenzo[e]indol-3-ium-2-yl]penta-2,4-dienylidene]-3-ethyl-1,1-dimethyl-8-sulfobenzo[e]indole-6-sulfonate Chemical compound CC1(C)C(C2=CC(=CC(=C2C=C2)S([O-])(=O)=O)S(O)(=O)=O)=C2N(CC)\C1=C/C=C/C=C/C(C(C1=C2C=C(C=C(C2=CC=C11)S(O)(=O)=O)S(O)(=O)=O)(C)C)=[N+]1CCCCCC(=O)ON1C(=O)CCC1=O DOMDXTIMIZCSNC-UHFFFAOYSA-N 0.000 description 2
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- XMIIGOLPHOKFCH-UHFFFAOYSA-N 3-phenylpropionic acid Chemical compound OC(=O)CCC1=CC=CC=C1 XMIIGOLPHOKFCH-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000000116 DAPI staining Methods 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 2
- 238000009509 drug development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 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 2
- 210000002216 heart Anatomy 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- SUMDYPCJJOFFON-UHFFFAOYSA-N isethionic acid Chemical compound OCCS(O)(=O)=O SUMDYPCJJOFFON-UHFFFAOYSA-N 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 238000002953 preparative HPLC Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- ZDPHROOEEOARMN-UHFFFAOYSA-N undecanoic acid Chemical compound CCCCCCCCCCC(O)=O ZDPHROOEEOARMN-UHFFFAOYSA-N 0.000 description 2
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical compound CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 2
- QBYIENPQHBMVBV-HFEGYEGKSA-N (2R)-2-hydroxy-2-phenylacetic acid Chemical compound O[C@@H](C(O)=O)c1ccccc1.O[C@@H](C(O)=O)c1ccccc1 QBYIENPQHBMVBV-HFEGYEGKSA-N 0.000 description 1
- MIOPJNTWMNEORI-GMSGAONNSA-N (S)-camphorsulfonic acid Chemical compound C1C[C@@]2(CS(O)(=O)=O)C(=O)C[C@@H]1C2(C)C MIOPJNTWMNEORI-GMSGAONNSA-N 0.000 description 1
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- YGTUPRIZNBMOFV-UHFFFAOYSA-N 2-(4-hydroxybenzoyl)benzoic acid Chemical compound OC(=O)C1=CC=CC=C1C(=O)C1=CC=C(O)C=C1 YGTUPRIZNBMOFV-UHFFFAOYSA-N 0.000 description 1
- WXHLLJAMBQLULT-UHFFFAOYSA-N 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-n-(2-methyl-6-sulfanylphenyl)-1,3-thiazole-5-carboxamide;hydrate Chemical compound O.C=1C(N2CCN(CCO)CC2)=NC(C)=NC=1NC(S1)=NC=C1C(=O)NC1=C(C)C=CC=C1S WXHLLJAMBQLULT-UHFFFAOYSA-N 0.000 description 1
- ALKYHXVLJMQRLQ-UHFFFAOYSA-N 3-Hydroxy-2-naphthoate Chemical compound C1=CC=C2C=C(O)C(C(=O)O)=CC2=C1 ALKYHXVLJMQRLQ-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- ZRPLANDPDWYOMZ-UHFFFAOYSA-N 3-cyclopentylpropionic acid Chemical compound OC(=O)CCC1CCCC1 ZRPLANDPDWYOMZ-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- AWQSAIIDOMEEOD-UHFFFAOYSA-N 5,5-Dimethyl-4-(3-oxobutyl)dihydro-2(3H)-furanone Chemical compound CC(=O)CCC1CC(=O)OC1(C)C AWQSAIIDOMEEOD-UHFFFAOYSA-N 0.000 description 1
- WDJHALXBUFZDSR-UHFFFAOYSA-N Acetoacetic acid Natural products CC(=O)CC(O)=O WDJHALXBUFZDSR-UHFFFAOYSA-N 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- LSPHULWDVZXLIL-UHFFFAOYSA-N Camphoric acid Natural products CC1(C)C(C(O)=O)CCC1(C)C(O)=O LSPHULWDVZXLIL-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 239000005639 Lauric acid Substances 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- IWYDHOAUDWTVEP-UHFFFAOYSA-N R-2-phenyl-2-hydroxyacetic acid Natural products OC(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-N 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical class 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 229960005261 aspartic acid Drugs 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 1
- 229940092714 benzenesulfonic acid Drugs 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 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 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- LSPHULWDVZXLIL-QUBYGPBYSA-N camphoric acid Chemical compound CC1(C)[C@H](C(O)=O)CC[C@]1(C)C(O)=O LSPHULWDVZXLIL-QUBYGPBYSA-N 0.000 description 1
- 150000001767 cationic compounds Chemical class 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- WOWBFOBYOAGEEA-UHFFFAOYSA-N diafenthiuron Chemical compound CC(C)C1=C(NC(=S)NC(C)(C)C)C(C(C)C)=CC(OC=2C=CC=CC=2)=C1 WOWBFOBYOAGEEA-UHFFFAOYSA-N 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- VFNGKCDDZUSWLR-UHFFFAOYSA-N disulfuric acid Chemical compound OS(=O)(=O)OS(O)(=O)=O VFNGKCDDZUSWLR-UHFFFAOYSA-N 0.000 description 1
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 1
- 230000002121 endocytic effect Effects 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 229960002598 fumaric acid Drugs 0.000 description 1
- 229950006191 gluconic acid Drugs 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 229940071870 hydroiodic acid Drugs 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 229910001411 inorganic cation Inorganic materials 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 229940098895 maleic acid Drugs 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 229960002510 mandelic acid Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- LNOPIUAQISRISI-UHFFFAOYSA-N n'-hydroxy-2-propan-2-ylsulfonylethanimidamide Chemical compound CC(C)S(=O)(=O)CC(N)=NO LNOPIUAQISRISI-UHFFFAOYSA-N 0.000 description 1
- YZMHQCWXYHARLS-UHFFFAOYSA-N naphthalene-1,2-disulfonic acid Chemical compound C1=CC=CC2=C(S(O)(=O)=O)C(S(=O)(=O)O)=CC=C21 YZMHQCWXYHARLS-UHFFFAOYSA-N 0.000 description 1
- KVBGVZZKJNLNJU-UHFFFAOYSA-N naphthalene-2-sulfonic acid Chemical compound C1=CC=CC2=CC(S(=O)(=O)O)=CC=C21 KVBGVZZKJNLNJU-UHFFFAOYSA-N 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 150000004968 peroxymonosulfuric acids Chemical class 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- IUGYQRQAERSCNH-UHFFFAOYSA-N pivalic acid Chemical compound CC(C)(C)C(O)=O IUGYQRQAERSCNH-UHFFFAOYSA-N 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 238000001946 ultra-performance liquid chromatography-mass spectrometry Methods 0.000 description 1
- 229940005605 valeric acid Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2827—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against B7 molecules, e.g. CD80, CD86
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/07—Tetrapeptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- 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
- A61K47/68—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6849—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
- C07K5/0205—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)3-C(=0)-, e.g. statine or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Cell Biology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本申请涉及生物制药及生物技术领域，特别涉及靶向B7‑H3抗体‑MMAE偶联物及其应用。本申请提供一种抗体‑药物偶联物，包括式Ⅰ所示的化合物或其药学上可接受的盐或溶剂合物：
Description
技术领域
本申请涉及生物制药及生物技术领域，特别涉及靶向B7-H3抗体-MMAE偶联物及其应用。
背景技术
B7-H3(CD276)，是一种跨膜蛋白，在血管生成和转移、肿瘤迁移侵袭以及肿瘤生长等中发挥着非常重要的作用。研究表明B7-H3在多种实体瘤中高表达，包括脑胶质瘤、非小细胞肺癌、肾细胞癌、乳腺癌以及前列腺癌等等，而在正常组织中B7-H3表达量低。因此B7-H3是一个合适的靶点用于抗癌药物研发。
抗体-药物偶联物(ADCs)由三部分组成：抗体、连接子以及高活性药物分子。通过抗体的靶向作用可以将高活性药物分子特异性的输送到肿瘤部位，从而起到高效低毒的作用，可用于肿瘤靶向治疗。目前已有14个ADCs被FDA被准上市，另外还有100多个ADCs处于临床研究。基于B7-H3的药物开发，目前已有抗体药物、双抗、CAR-T、小分子药物等多种手段，但到目前为止，尚未有针对该靶点的上市药物。靶向B7-H3的ADCs药物也开始逐渐兴起。
发明内容
为解决现有技术中的技术问题，本申请的目的在于提供靶向B7-H3抗体-MMAE偶联物及其应用。本发明提供一种通过桥联手段制备一种均一的靶向于B7-H3的抗体-药物偶联物(ADC)并将其用于在癌症靶向治疗中，尤其是脑胶质瘤的治疗中。
为实现上述目的及其他相关目的，本申请一方面提供一种抗体-药物偶联物，包括式Ⅰ所示的化合物或其药学上可接受的盐或溶剂合物：
其中，Anti-B7-H3为抗B7-H3抗体，D为药物，L为连接基团；n表示药物抗体比DAR，n为1～5的自然数。
在本申请的具体实方式中，抗体-药物偶联物选自SS-401，结构式为：
本申请另一方面提供一种抗体-药物偶联物的制备方法，包括以下步骤：
1)将化合物1、11-叠氮基-3,6,9-三氧代十一酸、N-甲基吗啉、1-(3-二甲胺基丙基)-3-乙基碳二亚胺和1-羟基苯并三氮唑以溶液形式反应，得到化合物2：
2)化合物2溶液中加入化合物3，无水硫酸铜和抗坏血酸钠，反应后过滤除去不溶物，分离得到化合物4：
3)将B7-H3单抗401溶液和三(2-羧乙基)膦溶液进行第一次振荡反应，再加入化合物4溶液进行第二次振荡反应，除去小分子化合物，得到抗体-药物偶联物SS-401：
本申请另一方面提供前述的抗体-药物偶联物在制备癌症靶向治疗产品或制备治疗增殖性疾病产品中的应用。
与现有技术相比，本申请的有益效果为：
1、本发明是基于双乙烯磺酰胺连接子技术，以及特异性靶向于B7-H3的抗体合成靶向于B7-H3的抗体-药物偶联物。
2、本发明基于抗体内在的硫-硫键断开后，再通过双乙烯磺酰胺连接子与药物桥联，可得到DAR为4左右的均一性好的偶联产物。
3、本发明提供的抗体-药物偶联物，在体内外具有很好的效果。
4、本发明提供了所制备的抗体药物偶联物在治疗高增殖性疾病或病灶的用途，尤其是在脑胶质瘤治疗中的用途。
附图说明
图1为本发明中涉及的典型的药物-连接子的制备路线及结构。
图2为本发明中涉及的ADC SS-401的HR-ESI-MS谱图。
图3为本发明中涉及的细胞U251,U87和U87/B7-H3 KO中B7-H3表达图。
图4为本发明中涉及的ADC SS-401、单抗401和MMAE对细胞U251的体外增殖抑制作用。
图5为本发明中涉及的ADC SS-401、单抗401和MMAE对细胞U87的体外增殖抑制作用。
图6为本发明中涉及的ADC SS-401、单抗401和MMAE对细胞U87/B7-H3 KO的体外增殖抑制作用。
图7为本发明中涉及的ADC SS-401和单抗401在不同浓度梯度下与细胞U87的亲和力。
图8为本发明中涉及的ADC SS-401和单抗401在细胞U87中的内吞效应。
图9为本发明中涉及的荧光标记偶联物Cy5.5-SS-401在脑胶质瘤异种移植瘤中的活体成像图。
图10为本发明中涉及的荧光标记偶联物Cy5.5-SS-401在脑胶质瘤异种移植瘤中，小鼠肿瘤部位、心脏、脾、肝、胃和肾脏的成像图。
图11为本发明中涉及的ADC SS-401和单抗401对脑胶质瘤U87异种移植瘤的抑制作用。
图12为本发明中涉及的ADC SS-401和单抗401对脑胶质瘤U87异种移植瘤小鼠体重的影响。
图13为本发明抗体-药物偶联物SS-401的结构图。
具体实施方式
为了使本申请的发明目的、技术方案和有益效果更加清晰，下面结合实施例对本申请作进一步说明。应理解，所述实施例只用于解释本申请，并非用于限定申请的范围。下述实施例中所使用的试验方法如无特殊说明，均为常规方法，熟悉此技术的人士可由本说明所揭露的内容容易地了解本申请的其他优点及功效。
本申请的发明人经过大量探索研究，发现了靶向B7-H3抗体-MMAE偶联物及其应用，在此基础上完成了本申请。
本申请一方面提供一种抗体-药物偶联物，包括式Ⅰ所示的化合物或其药学上可接受的盐或溶剂合物：
其中，Anti-B7-H3为抗B7-H3抗体，D为药物，L为连接基团；n表示药物抗体比DAR，n为1～5的自然数。药物抗体比(DAR)是指附着在单个单抗上的有效载荷分子的平均数量，通常在2到4个分子之间。本发明是基于双乙烯磺酰胺连接子技术，以及特异性靶向于B7-H3的抗体合成靶向于B7-H3的抗体-药物偶联物。
本发明中，术语“药学上可接受的盐”通常指当以适当的方式用于治疗时(特别是在人类和/或哺乳动物中应用或使用时)在生理学上可耐受的任何盐(通常来说，这意味着它是无毒的，特别是作为抗衡离子的结果是无毒的)。这些生理上可接受的盐可以是与阳离子或碱形成的，并且在本发明的上下文中，尤其是在人类和/或哺乳动物中施用时，它们应该被理解为由按照本发明所提供的至少一种化合物，通常为酸(去质子化的)，如阴离子和至少一种生理学上耐受的阳离子(优选无机阳离子)形成的盐。在本发明的上下文中，具体地可以包括与碱金属和碱土金属形成的盐、以及与铵阳离子(NH4 +)形成的盐，具体可以是包括但不限于与(单)或(二)钠、(单)或(二)钾、镁或钙形成的盐。这些生理上可接受的盐也可以是与阴离子或酸形成的，并且在本发明的上下文中，特别是在人类和/或哺乳动物中施用时，它们应该被理解为由按照本发明所提供的至少一种化合物，通常质子化的(例如在氮上)，如阳离子和至少一种生理上可耐受的阴离子形成的盐。在本发明的上下文中，具体地可以包括由生理上可耐受的酸形成的盐，即特定的活性化合物与生理上可耐受的有机或无机酸形成的盐，具体可以是包括但不限于与盐酸、氢溴酸、氢碘酸；硫酸、过硫酸、焦硫酸；磷酸、硝酸；甲磺酸、乙磺酸、苯磺酸、对甲苯磺酸、三氟甲磺酸；甲酸、乙酸、乙酰乙酸、三氟乙酸、丙酮酸、丙酸、丁酸、戊酸、己酸、庚酸、十一烷酸、月桂酸；苯甲酸、水杨酸、2-(4-羟基苯甲酰基)苯甲酸、樟脑酸、肉桂酸、环戊烷丙酸、二葡萄糖酸、3-羟基-2-萘甲酸、烟酸、果胶酸、3-苯基丙酸、苦味酸、特戊酸；2-羟基乙磺酸、氨基磺酸、十二烷基硫酸、2-萘磺酸、萘二磺酸、樟脑磺酸；柠檬酸、酒石酸、硬脂酸、乳酸、草酸、丙二酸、琥珀酸、苹果酸、肥酸、藻酸、马来酸、富马酸、D-葡萄糖酸、扁桃酸、抗坏血酸、天冬氨酸、磺基水杨酸形成的盐。
在本文中，术语“溶剂合物”是指本公开的化合物或其药学上可接受的盐，其包含在非共价分子之间通过力结合的化学计量或非化学计量的溶剂。优选的溶剂是挥发性和无毒的，并且可以以非常小的剂量给予人。溶剂的实例但不限于包括水、异丙醇、乙醇、甲醇、DMSO、乙酸乙酯、乙酸和乙醇胺。术语“水合物”是指溶剂分子是水的络合物。
本申请提供的抗体-药物偶联物中，L选自羧基、胺基、炔基活叠氮中的一种或多种的组合。
本申请提供的抗体-药物偶联物中，n为4。
本申请提供的抗体-药物偶联物中，Anti-B7-H3经内在的二硫键通过连接基团与药物相连。抗B7-H3抗体为抗B7-H3单克隆抗体。在本申请一具体实施例中，Anti-B7-H3为401(专利202210020231.3)。
本申请提供的抗体-药物偶联物中，药物选自一甲基澳瑞他汀E(MMAE)、一甲基澳瑞他汀F(MMAF)、康普瑞汀A、阿糖胞苷、吉西他滨、美登素、DM1、DM4、卡奇霉素及其衍生物、阿霉素、倍癌霉素及其衍生物、吡咯并苯并二氮杂卓(PBD)、SN-38、喜树碱及其类似物、α-安曼丁以及微管蛋白裂解素类似物。在本申请一具体实施例中，药物选自MMAE。
本申请提供的抗体-药物偶联物中，抗体-药物偶联物的结构式为：
也可以表示成：
也可以表示成图13所示的结构。
本申请另一方面提供前述的抗体-药物偶联物的制备方法，包括以下步骤：
1)将化合物1、11-叠氮基-3,6,9-三氧代十一酸、N-甲基吗啉(NMM)、1-(3-二甲胺基丙基)-3-乙基碳二亚胺(EDCl)和1-羟基苯并三氮唑(HOBT)以溶液形式反应，得到化合物2：
2)化合物2溶液中加入化合物3，无水硫酸铜和抗坏血酸钠，反应后过滤除去不溶物，分离得到化合物4：
3)将B7-H3单抗401溶液和三(2-羧乙基)膦溶液进行第一次振荡反应，再加入化合物4溶液进行第二次振荡反应，除去小分子化合物，得到抗体-药物偶联物：
本申请提供的制备方法中，步骤1)是指将化合物1、11-叠氮基-3,6,9-三氧代十一酸、NMM、EDCl和HOBT以溶液形式反应，得到化合物2。其中，溶液的溶剂为N,N-二甲基甲酰胺(DMF)。反应的温度为室温，反应的时间为2～24h；具体地，为2～10h、10～12h、12～16h、或16～24h等。
本申请提供的制备方法中，步骤2)是指化合物2溶液中加入化合物3，无水硫酸铜和抗坏血酸钠，反应后过滤除去不溶物，分离得到化合物4。其中，化合物2溶液的溶剂为叔丁醇(tBuOH)、H2O和DMF的混合溶液。在本申请一具体实施例中，tBuOH、H2O和DMF的体积比为1：1：1。反应的温度为室温，反应的时间为1～4h；具体地，为1～2h、2～3h、或2～3h等。分离是通过HPLC实现。HPLC的条件为：固定相：C-18柱，流动相：CH3CN/H2O＝10～100％，共30分钟。
本申请提供的制备方法中，步骤3)是指将B7-H3单抗溶液401和三(2-羧乙基)膦溶液进行第一次振荡反应，再加入化合物4溶液进行第二次振荡反应，除去小分子化合物，得到抗体-药物偶联物。其中，步骤3)是在微孔板中进行。B7-H3单抗溶液401的溶剂为磷酸盐缓冲溶液(PBS)，pH＝7.4。三(2-羧乙基)膦溶液的溶剂为纯水，pH为7.0。第一次振荡反应的反应温度为室温，反应时间为0.5～2h；具体地，为0.5～1h、1～1.5h、或1.5～2h等。化合物4溶液的溶剂为二甲基亚砜(DMSO)，第二次振荡反应的反应温度为室温，反应时间为2～18h；具体地，可以为2～6h、6～12h、或12～18h等。除去小分子化合物通过Zeba脱盐柱(ZebaTMSpin Desalting Columns,7K MWCO,0.5mL)进行。本发明基于抗体内在的硫-硫键断开后，再通过双乙烯磺酰胺连接子与药物桥联，可得到DAR为4左右的均一性好的偶联产物SS-401。
本申请另一方面提供前述的抗体-药物偶联物在制备癌症靶向治疗产品或制备治疗增殖性疾病产品中的应用。
本申请提供的应用中，癌症靶向治疗的靶点为B7-H3。增殖性疾病的特征为细胞生成一种抗原或靶点，抗原能与B7-H3的抗体特异性结合。进一步地，增殖性疾病为B7-H3表达的病灶。更进一步地，增殖性疾病为脑胶质瘤。本发明提供了所制备的抗体药物偶联物在在体内外具有很好的效果，可应用于治疗高增殖性疾病或病灶，尤其是治疗脑胶质瘤。
下面通过实施例对本申请予以进一步说明，但并不因此而限制本申请的范围。
实施例1
抗体-药物偶联物的制备
制备路线如图1所示，过程如下：
将化合物1(参考文献Bioorganic Chemistry 134(2023)106463)(100mg，0.139mmol)、11-叠氮基-3,6,9-三氧代十一酸(35.58mg，0.153mmol)、N-甲基吗啉NMM(140.6mg，1.39mmol)、EDCl(53.48mg，0.279mmol)和HOBt(37.92mg,0.279mmol)溶于DMF(5mL)中，室温下反应12h。用制备HPLC(固定相为C-18硅胶柱，流动相为CH3CN/H2O＝10～100％，30分钟)分离得到产物白色固体2(111mg)。
将化合物2(6mg,0.015mmol)溶解于1.5mL的tBuOH/H2O/DMF(1/1/1)混合溶液中，加入化合物3(20mg,0.015mmol)，无水硫酸铜(2mg,0.015mmol)和抗坏血酸钠(4mg,0.022mmol)。混合物在室温反应2小时后，过滤除去不溶物，用制备HPLC(固定相：C-18柱，流动相：CH3CN/H2O＝10～100％，共30分钟)分离得到白色固体4共13mg。.1H NMR(500MHz,DMSO)δ10.13–10.06(m,1H),8.44(q,J＝6.6,5.9Hz,1H),8.35(dd,J＝17.4,9.8Hz,2H),7.61–7.55(m,2H),7.39–7.22(m,8H),7.20–7.13(m,1H),6.83–6.67(m,2H),6.20–6.01(m,5H),5.03(ddd,J＝33.8,26.7,13.5Hz,2H),4.51–4.11(m,12H),3.95(qd,J＝18.4,15.8,4.9Hz,6H),3.76(dt,J＝11.6,5.5Hz,3H),3.58–3.49(m,10H),3.26–3.08(m,11H),3.07–2.82(m,10H),2.68(td,J＝11.1,10.5,5.3Hz,1H),2.15–1.90(m,4H),1.83–1.62(m,3H),1.63–1.40(m,3H),1.39–1.21(m,2H),1.10–0.96(m,6H),0.90–0.71(m,29H).13C NMR(151MHz,DMSO)δ10.3,14.0,15.0,15.3,15.5,15.9,17.9,18.0,18.5,19.2,22.1,23.1,24.3,25.3,26.8,28.7,29.0,29.3,29.9,31.1,31.3,33.3,34.5,40.4,41.8,43.2,43.7,44.4,44.6,45.0,46.2,47.1,47.2,47.5,49.2,49.4,49.8,53.2,54.1,54.2,54.4,56.6,57.2,58.2,58.7,60.3,60.9,67.9,68.7,69.6,69.6,69.7,70.3,74.8,81.6,85.4,119.0,123.0,126.4,126.5,126.7,126.7,127.4,127.6,127.7,127.8,128.2,129.7,129.8,132.6,132.7,135.6,135.6,143.7,144.5,158.9,168.0,168.1,168.8,169.0,170.5,170.7,172.4.ESI-HRMS calculated for C78H124N16O21S2[M+H]+:1684.8568,Found:1684.8512.
将化合物4溶于DMSO，配置成10mmol/L的溶液；401单抗溶于PBS(25mmol/L Tris，pH 7.4)缓冲溶液，配成3mg/mL的溶液；TCEP溶于水配成10mM的溶液，用NaOH/H3PO4调节pH为7.0。在330μL的96孔板中加入100μL 401单抗溶液，0.83μ的TCEP溶液，在微孔板振荡器上室温下孵育1h。随后加入1.67μL上述制备的化合物4，然后放在微孔板振荡器上室温下反应12小时后，通过Zeba脱盐柱(ZebaTM Spin Desalting Columns,7K MWCO,0.5mL)除去过量的小分子化合物。得到相应的抗体-药物偶联物ADC(SS-401)。用UPLC-MS(型号ABsciex 4600)分析反应结果。
实施例2
细胞B7-H3抗原表达检测
配制500mLFACS缓冲液(PBS+0.1％BSA)，将U87(人脑胶质瘤细胞)、U87-B7-H3-KO(在U87细胞中用常规技术手段敲除B7-H3基因)、U251(人胶质瘤细胞)细胞收集至15毫升离心管中，用FACS缓冲液洗涤后重悬细胞并计数分装到1.5毫升离心管中(保证每管活细胞数目充足，且每种细胞均设置三重复)，再次离心沉淀细胞；然后将所有细胞用浓度为100nM的200μL体积的FITC anti-Human CD276(B7-H3)溶液于冰上孵育30min。孵育结束后用FACS清洗2次，每个样品用200μL且浓度为1μg/mL的DAPI溶液重悬，用CytoFLEX Flow Cytometer流式细胞仪测量FITC的荧光强度。使用FlowJo 10软件分析处理数据。
结果如图3所示。
实施例3
细胞增殖抑制检测
将处于对数生长期的细胞接种于96孔板中，每孔培养基体积为100μL。对于悬浮细胞，每孔接种细胞2×104个，铺板后可直接加药孵育48h后处理；而对于贴壁细胞，每孔3×103个，需放置在培养箱中培养24h等待细胞完全贴壁后再加药。将实施例1制备得到的ADC(SS-401)，单抗401以及小分子毒性物质MMAE(毕得医药)以一定浓度梯度进行稀释后，加入细胞板孵育72小时，每个浓度设置3复孔，并且另外设置溶媒对照组(只铺细胞但不加药)以及空白对照组(只加等体积培养基)。药物处理结束后通过CCK-8试剂检测活细胞，并用酶标仪读取450nm波长的吸光度(OD值)。通过公式计算抑制率(％)＝(OD对照-OD给药)/(OD对照-OD空白)×100％。注意：细胞接种浓度及CCK8显色时间均需要摸索，最后显色时间保证OD值在1.0-1.5。
结果如图4～6所示。
实施例4
抗体-药物偶联物的亲和力作用测定
在此实验过程中所有样品的离心以及孵育过程均需保持在4℃的环境下(置于低温离心机中离心或冰上孵育)。
1、细胞收集：使用胰酶消化收集生长状态良好的U87细胞并转移至15mL离心管中，用FACS缓冲液洗涤，再次离心细胞并将其重悬计数后以每个离心管5×105个活细胞数分装至1.5mL离心管，2000rpm离心3min，去上清；
2、抗体/ADCs结合细胞：用PBS溶液稀释单抗401或者ADC(SS-401)，用不同浓度(0.01、0.03、0.1、0.3、1、3、10、30、100、300nM)的200μL单抗401或者SS-401溶液重悬细胞后形成细胞悬液，置于冰上孵育30min，在孵育期间需要对细胞进行多次吹吸防止细胞沉底；
3、洗涤：使用FACS缓冲液重复洗涤细胞两次；
4、孵育二抗：在每个样品中加入200μL体积以1:200比例稀释的二抗溶液GoatAnti-Human IgG H&L(650)，于冰上避光孵育30min；
4、洗涤：使用FACS缓冲液洗涤细胞两次；
6、DAPI染色并上机检测：用DAPI溶液(浓度为1μg/mL)重悬，15分钟后使用流式细胞仪进行检测。
结果如图7所示。
实施例5
抗体-药物偶联物内吞作用测定
在此实验过程中所有4℃对照组样品的离心以及孵育过程均需保持在4℃的环境下(置于低温离心机中离心或冰上孵育)。
1、细胞收集：按照贴壁细胞处理方式用胰酶消化收集生长状态良好的U87细胞并转移到15mL离心管，用FACS缓冲液洗涤；再次离心细胞并重悬计数后以每个离心管5×105个活细胞数分装至1.5mL离心管，2000rpm离心3min，去上清；设置4℃对照组以及37℃实验组。
2、抗体/ADCs结合细胞：用PBS溶液稀释单抗401或者SS-401，用100nM的200μL单抗401或者SS-401溶液重悬细胞后形成细胞悬液，置于冰上孵育30min，在孵育期间需要对细胞进行多次吹吸防止细胞沉底；
3、洗涤：使用FACS缓冲液重复洗涤细胞两次；
4、内化过程：向各离心管内加入200μL新鲜的完全培养基重悬细胞后，将37℃组转移至37℃恒温培养箱中开盖孵育3个小时，使结合于细胞表面的单抗401或者SS-401内化；
并将4℃组置于冰上孵育相同时间作为对照；
5、终止内化：将37℃组转移至冰上，并添加200μL冷PBS溶液终止内化。同时向4℃组加入等体积冷PBS，以2000rpm于4℃下离心3min后弃去上清液，并用FACS缓冲液洗涤一次；
6、孵育二抗：每个样品加入200μL 1:200稀释的二抗Goat Anti-Human IgG H&L(
650)，于冰上避光孵育30min；
7、洗涤未结合二抗：使用FACS缓冲液重复洗涤细胞两次；
8、DAPI染色并上机检测：用DAPI溶液(浓度为1μg/mL)重悬，15分钟后使用流式细胞仪进行检测。
内化效率计算公式：内化效率(％)＝(4℃组荧光强度-37℃组荧光强度)/4℃组荧光强度×100％。
结果如8所示。
实施例6
抗体-药物偶联物体内药效检测
采购SPF级别5周龄BALB/c裸鼠雄性小鼠，置于国家蛋白质中心SPF级动物房中饲养并适应环境一周，期间观察小鼠的身体情况及精神状态。同时复苏并扩增U87细胞，待细胞数目满足实验接种需求并且生长状态良好时用胰酶进行消化，然后使用PBS洗涤2次后重悬计数，并用1:2稀释的ABW基质胶调整细胞密度至1.5×107/mL。在小鼠后肢右侧腹股沟上方部血管丰富处给每只小鼠皮下接种3×106个细胞(200μL)，注射时需要多次轻吹细胞进行混匀。接种后每隔四天对小鼠肿瘤体积大小以及体重变化进行测量监测(使用游标卡尺及电子天平测量)。肿瘤体积大小公式为：肿瘤体积(mm3)＝长(mm)×宽2(mm2)×0.5。当观察到小鼠平均肿瘤体积长至约150mm3左右时，将满足标准的成瘤小鼠按照实验需求进行随机分组使各组小鼠的平均肿瘤体积接近。之后进行尾静脉注射治疗，依照实验设计选择适宜给药方案，本实验中尾静脉注射给药剂量主要分为2mg·kg-1和6mg·kg-1，并且给药体积保持200μL；将第一次给药记为第0天，在第0、4或7天各给药一次，连续给药2至3次，期间需每隔两天监测并记录肿瘤体积及小鼠体重变化，实验结束后对小鼠进行安乐死。
结果如图11～12所示。
实施例7
抗体-药物偶联物体内荧光成像检测
采用小鼠活体成像技术在U87异种移植模型中评估ADCs的肿瘤靶向能力及其在小鼠体内的分布代谢。使用Cy5.5 NHS酯将SS-401进行染色标记，当小鼠肿瘤体积生长至适宜大小时，将其随机分成两组，每组四只，用Cy5.5 NHS酯标记的SS-401以6mg·kg-1的剂量对小鼠进行尾静脉注射给药，并在给药结束后第24、48、72小时的时间点麻醉小鼠，放入小动物化学荧光成像系统中进行成像，观察不同时间点的荧光信号分布变化(每次成像时小鼠位置需保持一致)。并且在第72h后解剖小鼠，取出其心、肝、脾、肺、肾以及肿瘤组织进行局部成像以观察其器官荧光信号分布差异。使用Living Image软件对图像进行分析定量。
结果如图9～10所示。
上述实施例仅例示性说明本发明的原理及其功效，而非用于限制本申请。任何熟悉此技术的人士皆可在不违背本申请的精神及范畴下，对上述实施例进行修饰或改变。因此，举凡所属技术领域中具有通常知识者在未脱离本发明所揭示的精神与技术思想下所完成的一切等效修饰或改变，仍应由本申请的权利要求所涵盖。
Claims (10)
1.一种抗体-药物偶联物，其特征在于，包括式Ⅰ所示的化合物或其药学上可接受的盐或溶剂合物：
其中，Anti-B7-H3为抗B7-H3抗体，D为药物，L为连接基团；n表示药物抗体比DAR，n为1～5的自然数。
2.如权利要求1所述的抗体-药物偶联物，其特征在于，所述L选自羧基、胺基、炔基、或叠氮中的一种或多种的组合；
和/或，所述n为4；
和/或，所述Anti-B7-H3经内在的二硫键通过连接基团与药物相连；
和/或，所述抗B7-H3抗体为抗B7-H3单克隆抗体；
和/或，药物选自一甲基澳瑞他汀E、一甲基澳瑞他汀F、康普瑞汀A、阿糖胞苷、吉西他滨、美登素、DM1、DM4、卡奇霉素及其衍生物、阿霉素、倍癌霉素及其衍生物、吡咯并苯并二氮杂卓(PBD)、SN-38、喜树碱类似物、α-安曼丁以及微管蛋白裂解素类似物。
3.如权利要求2所述的抗体-药物偶联物，其特征在于，所述抗体-药物偶联物选自SS-401，结构式为：
4.如权利要求3所述的抗体-药物偶联物的制备方法，其特征在于，包括以下步骤：
1)将化合物1、11-叠氮基-3,6,9-三氧代十一酸、N-甲基吗啉、1-(3-二甲胺基丙基)-3-乙基碳二亚胺和1-羟基苯并三氮唑以溶液形式反应，得到化合物2：
2)化合物2溶液中加入化合物3，无水硫酸铜和抗坏血酸钠，反应后过滤除去不溶物，分离得到化合物4：
3)将B7-H3单抗401溶液和三(2-羧乙基)膦溶液进行第一次振荡反应，再加入化合物4溶液进行第二次振荡反应，除去小分子化合物，得到抗体-药物偶联物SS-401：
5.如权利要求4所述的制备方法，其特征在于，步骤1)中，所述溶液的溶剂为N,N-二甲基甲酰胺；所述反应的温度为室温，所述反应的时间为2～24h。
6.如权利要求4所述的制备方法，其特征在于，步骤2)中，所述化合物2溶液的溶剂为N,N-二甲基甲酰胺、叔丁醇和蒸馏水的体积比为1:1:1的混合溶液；
和/或，步骤2)中，所述反应的温度为室温；
和/或，步骤2)中，所述反应的时间为1～4h。
7.如权利要求4所述的制备方法，其特征在于，步骤3)是在微孔板中进行；B7-H3单抗401溶液的溶剂为磷酸盐缓冲溶液；三(2-羧乙基)膦溶液的溶剂为纯水，化合物4溶液的溶剂为二甲基亚砜；
和/或，步骤3)中，第一次振荡反应的反应温度为室温，反应时间为0.5～2h；
和/或，步骤3)中，第二次振荡反应的反应温度为室温，反应时间为2～18h；
和/或，步骤3)中，小分子化合物的分子量≤7000。
8.如权利要求1～3任一所述的抗体-药物偶联物在制备癌症靶向治疗产品或制备治疗增殖性疾病产品中的应用。
9.如权利要求8所述的应用，其特征在于，所述癌症靶向治疗的靶点为B7-H3。
10.如权利要求8所述的应用，其特征在于，所述增殖性疾病的特征为细胞生成一种抗原或靶点，抗原能与B7-H3的抗体特异性结合；优选地，所述的增殖性疾病为B7-H3表达的病灶；更优选地，所述增殖性疾病为脑胶质瘤。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310475213.9A CN116747317A (zh) | 2023-04-27 | 2023-04-27 | 靶向b7-h3抗体-mmae偶联物及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310475213.9A CN116747317A (zh) | 2023-04-27 | 2023-04-27 | 靶向b7-h3抗体-mmae偶联物及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116747317A true CN116747317A (zh) | 2023-09-15 |
Family
ID=87950321
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310475213.9A Pending CN116747317A (zh) | 2023-04-27 | 2023-04-27 | 靶向b7-h3抗体-mmae偶联物及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116747317A (zh) |
-
2023
- 2023-04-27 CN CN202310475213.9A patent/CN116747317A/zh active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10342846B2 (en) | Nanoparticle drug delivery systems | |
| JP7103953B2 (ja) | 薬物としてアマトキシンの誘導体を有する抗体薬物コンジュゲート | |
| CN114456186A (zh) | 一种喜树碱类衍生物及其配体-药物偶联物 | |
| JP2002537347A (ja) | 癌の標的付けられた治療のためのテキサン誘導体 | |
| EP2913064A1 (en) | Branched drug-linker conjugates for the coupling to biological targeting molecules | |
| WO2015038493A1 (en) | Synthesis of novel asymmetric bow-tie pamam dendrimer-based conjugates for tumor-targeting drug delivery | |
| CN108524919A (zh) | 用于改进的药物递送的载体 | |
| WO2020035027A1 (zh) | 连接子、含连接子的抗体偶联药物及连接子的用途 | |
| ES2701077T3 (es) | Conjugado de oligómero de ácido biliar para un nuevo transporte vesicular y uso del mismo | |
| JP2002518405A (ja) | ビタミンｂ１２誘導体及びそれらの製造方法 | |
| CN107335060A (zh) | 一类基于rgd多肽-化疗药物的小分子偶联物及其纳米前药系统 | |
| CN111001012A (zh) | 一种亲水碳酸酯型抗体偶联药物 | |
| US20220072147A1 (en) | Compound for preparation of antibody-payload conjugate and use thereof | |
| IL303707A (en) | Camptothecin antibody-drug conjugates and methods of using them | |
| TW202216724A (zh) | 一種喜樹鹼類藥物及其抗體偶聯物 | |
| US7314956B2 (en) | Multifunctional carrier for the delivery of a pharmacological agent or genetic material into a cell | |
| CN110759940A (zh) | 连接子、含连接子的抗体偶联药物及连接子的用途 | |
| Wei et al. | Site-specific construction of triptolide-based antibody-drug conjugates | |
| CN114053426A (zh) | 一种双药链接组装单元及双药靶向接头-药物偶联物 | |
| CN116747317A (zh) | 靶向b7-h3抗体-mmae偶联物及其应用 | |
| CN110507826B (zh) | 一种基于巨噬细胞的活细胞载药系统、其制备方法和应用 | |
| JP7153082B2 (ja) | システイン改変抗体－毒素複合体（ｔｄｃ）の部位特異的結合サイトのスクリーニング | |
| CN113713117B (zh) | 一种白蛋白结合型肿瘤环境响应型抗肿瘤前体药物及其制备方法和应用 | |
| CN116036303A (zh) | 一种抗体-药物偶联物及其制备方法和应用 | |
| WO2021213492A1 (zh) | 一类载药的大分子及其制备方法 |
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 |