CN116840467A - Method for preparing conjugate - Google Patents
Method for preparing conjugate Download PDFInfo
- Publication number
- CN116840467A CN116840467A CN202310810455.9A CN202310810455A CN116840467A CN 116840467 A CN116840467 A CN 116840467A CN 202310810455 A CN202310810455 A CN 202310810455A CN 116840467 A CN116840467 A CN 116840467A
- Authority
- CN
- China
- Prior art keywords
- glucose
- digoxin
- phosphate dehydrogenase
- mutant
- buffer
- 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
- 238000000034 method Methods 0.000 title claims abstract description 13
- 102100031126 6-phosphogluconolactonase Human genes 0.000 claims abstract description 32
- 108010029731 6-phosphogluconolactonase Proteins 0.000 claims abstract description 32
- 108010018962 Glucosephosphate Dehydrogenase Proteins 0.000 claims abstract description 32
- 230000035772 mutation Effects 0.000 claims abstract description 9
- LTMHDMANZUZIPE-PUGKRICDSA-N digoxin Chemical class C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)[C@H](O)C5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O LTMHDMANZUZIPE-PUGKRICDSA-N 0.000 claims description 58
- 239000000872 buffer Substances 0.000 claims description 22
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 12
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 9
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 4
- 208000034953 Twin anemia-polycythemia sequence Diseases 0.000 claims description 4
- 239000000010 aprotic solvent Substances 0.000 claims description 3
- RZQXOGQSPBYUKH-UHFFFAOYSA-N 3-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]azaniumyl]-2-hydroxypropane-1-sulfonate Chemical compound OCC(CO)(CO)NCC(O)CS(O)(=O)=O RZQXOGQSPBYUKH-UHFFFAOYSA-N 0.000 claims description 2
- 108700016232 Arg(2)-Sar(4)- dermorphin (1-4) Proteins 0.000 claims description 2
- 239000007983 Tris buffer Substances 0.000 claims description 2
- 238000011033 desalting Methods 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 21
- 238000002360 preparation method Methods 0.000 abstract description 7
- 238000011002 quantification Methods 0.000 abstract 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 229960005156 digoxin Drugs 0.000 description 49
- LTMHDMANZUZIPE-AMTYYWEZSA-N Digoxin Natural products O([C@H]1[C@H](C)O[C@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@](C)([C@H](O)C4)[C@H](C4=CC(=O)OC4)CC5)CC3)CC2)C[C@@H]1O)[C@H]1O[C@H](C)[C@@H](O[C@H]2O[C@@H](C)[C@H](O)[C@@H](O)C2)[C@@H](O)C1 LTMHDMANZUZIPE-AMTYYWEZSA-N 0.000 description 39
- LTMHDMANZUZIPE-UHFFFAOYSA-N digoxine Natural products C1C(O)C(O)C(C)OC1OC1C(C)OC(OC2C(OC(OC3CC4C(C5C(C6(CCC(C6(C)C(O)C5)C=5COC(=O)C=5)O)CC4)(C)CC3)CC2O)C)CC1O LTMHDMANZUZIPE-UHFFFAOYSA-N 0.000 description 39
- 239000003153 chemical reaction reagent Substances 0.000 description 25
- 102000004190 Enzymes Human genes 0.000 description 21
- 108090000790 Enzymes Proteins 0.000 description 21
- 239000000427 antigen Substances 0.000 description 13
- 102000036639 antigens Human genes 0.000 description 13
- 108091007433 antigens Proteins 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 10
- 238000010168 coupling process Methods 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 7
- 230000008878 coupling Effects 0.000 description 7
- 238000005859 coupling reaction Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 102000010705 glucose-6-phosphate dehydrogenase activity proteins Human genes 0.000 description 7
- 108040005050 glucose-6-phosphate dehydrogenase activity proteins Proteins 0.000 description 7
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 6
- 238000003018 immunoassay Methods 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 229940126586 small molecule drug Drugs 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- BAWFJGJZGIEFAR-NNYOXOHSSA-O NAD(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-O 0.000 description 4
- 238000003149 assay kit Methods 0.000 description 4
- 229940098773 bovine serum albumin Drugs 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 229940088597 hormone Drugs 0.000 description 4
- 239000005556 hormone Substances 0.000 description 4
- 230000002163 immunogen Effects 0.000 description 4
- 239000004816 latex Substances 0.000 description 4
- 229920000126 latex Polymers 0.000 description 4
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 4
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 4
- 239000003755 preservative agent Substances 0.000 description 4
- 230000002335 preservative effect Effects 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 230000000890 antigenic effect Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- -1 bromoacetyl Chemical group 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 238000003908 quality control method Methods 0.000 description 3
- 102200061299 rs1064794096 Human genes 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 229940035722 triiodothyronine Drugs 0.000 description 3
- 238000004879 turbidimetry Methods 0.000 description 3
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 101100013805 Aspergillus niger gsdA gene Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 206010003671 Atrioventricular Block Diseases 0.000 description 2
- NBSCHQHZLSJFNQ-GASJEMHNSA-N D-Glucose 6-phosphate Chemical compound OC1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H](O)[C@H]1O NBSCHQHZLSJFNQ-GASJEMHNSA-N 0.000 description 2
- XUIIKFGFIJCVMT-GFCCVEGCSA-N D-thyroxine Chemical compound IC1=CC(C[C@@H](N)C(O)=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-GFCCVEGCSA-N 0.000 description 2
- 101710088194 Dehydrogenase Proteins 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- VFRROHXSMXFLSN-UHFFFAOYSA-N Glc6P Natural products OP(=O)(O)OCC(O)C(O)C(O)C(O)C=O VFRROHXSMXFLSN-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- 206010019280 Heart failures Diseases 0.000 description 2
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102400000064 Neuropeptide Y Human genes 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- AUYYCJSJGJYCDS-LBPRGKRZSA-N Thyrolar Chemical compound IC1=CC(C[C@H](N)C(O)=O)=CC(I)=C1OC1=CC=C(O)C(I)=C1 AUYYCJSJGJYCDS-LBPRGKRZSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 230000004520 agglutination Effects 0.000 description 2
- 150000001540 azides Chemical class 0.000 description 2
- 229940097217 cardiac glycoside Drugs 0.000 description 2
- 239000002368 cardiac glycoside Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 229940125782 compound 2 Drugs 0.000 description 2
- 229940125898 compound 5 Drugs 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 229940045189 glucose-6-phosphate Drugs 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- LPMXVESGRSUGHW-HBYQJFLCSA-N ouabain Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1C[C@@]2(O)CC[C@H]3[C@@]4(O)CC[C@H](C=5COC(=O)C=5)[C@@]4(C)C[C@@H](O)[C@@H]3[C@@]2(CO)[C@H](O)C1 LPMXVESGRSUGHW-HBYQJFLCSA-N 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 229930002534 steroid glycoside Natural products 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- ZFXYFBGIUFBOJW-UHFFFAOYSA-N theophylline Chemical compound O=C1N(C)C(=O)N(C)C2=C1NC=N2 ZFXYFBGIUFBOJW-UHFFFAOYSA-N 0.000 description 2
- 229940034208 thyroxine Drugs 0.000 description 2
- XUIIKFGFIJCVMT-UHFFFAOYSA-N thyroxine-binding globulin Natural products IC1=CC(CC([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-UHFFFAOYSA-N 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- DNXHEGUUPJUMQT-UHFFFAOYSA-N (+)-estrone Natural products OC1=CC=C2C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 DNXHEGUUPJUMQT-UHFFFAOYSA-N 0.000 description 1
- UIKROCXWUNQSPJ-VIFPVBQESA-N (-)-cotinine Chemical compound C1CC(=O)N(C)[C@@H]1C1=CC=CN=C1 UIKROCXWUNQSPJ-VIFPVBQESA-N 0.000 description 1
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 description 1
- PROQIPRRNZUXQM-UHFFFAOYSA-N (16alpha,17betaOH)-Estra-1,3,5(10)-triene-3,16,17-triol Natural products OC1=CC=C2C3CCC(C)(C(C(O)C4)O)C4C3CCC2=C1 PROQIPRRNZUXQM-UHFFFAOYSA-N 0.000 description 1
- 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
- PUWBXDQHRRUKNY-CKOVTTQDSA-N (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal phosphoric acid Chemical compound OP(O)(O)=O.OP(O)(O)=O.OP(O)(O)=O.OP(O)(O)=O.OP(O)(O)=O.OP(O)(O)=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O PUWBXDQHRRUKNY-CKOVTTQDSA-N 0.000 description 1
- JWUBBDSIWDLEOM-XHQRYOPUSA-N (3e)-3-[(2e)-2-[1-(6-hydroxy-6-methylheptan-2-yl)-7a-methyl-2,3,3a,5,6,7-hexahydro-1h-inden-4-ylidene]ethylidene]-4-methylidenecyclohexan-1-ol Chemical compound C1CCC2(C)C(C(CCCC(C)(C)O)C)CCC2\C1=C\C=C1/CC(O)CCC1=C JWUBBDSIWDLEOM-XHQRYOPUSA-N 0.000 description 1
- FMNQRUKVXAQEAZ-JNRFBPFXSA-N (5z,8s,9r,10e,12s)-9,12-dihydroxy-8-[(1s)-1-hydroxy-3-oxopropyl]heptadeca-5,10-dienoic acid Chemical compound CCCCC[C@H](O)\C=C\[C@@H](O)[C@H]([C@@H](O)CC=O)C\C=C/CCCC(O)=O FMNQRUKVXAQEAZ-JNRFBPFXSA-N 0.000 description 1
- UCTWMZQNUQWSLP-VIFPVBQESA-N (R)-adrenaline Chemical compound CNC[C@H](O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-VIFPVBQESA-N 0.000 description 1
- 229930182837 (R)-adrenaline Natural products 0.000 description 1
- GMRQFYUYWCNGIN-ZVUFCXRFSA-N 1,25-dihydroxy vitamin D3 Chemical compound C1([C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CCCC(C)(C)O)C)=CC=C1C[C@@H](O)C[C@H](O)C1=C GMRQFYUYWCNGIN-ZVUFCXRFSA-N 0.000 description 1
- NVKAWKQGWWIWPM-ABEVXSGRSA-N 17-β-hydroxy-5-α-Androstan-3-one Chemical compound C1C(=O)CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 NVKAWKQGWWIWPM-ABEVXSGRSA-N 0.000 description 1
- DBPWSSGDRRHUNT-UHFFFAOYSA-N 17alpha-hydroxy progesterone Natural products C1CC2=CC(=O)CCC2(C)C2C1C1CCC(C(=O)C)(O)C1(C)CC2 DBPWSSGDRRHUNT-UHFFFAOYSA-N 0.000 description 1
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 1
- SFRDXVJWXWOTEW-UHFFFAOYSA-N 2-(hydroxymethyl)propane-1,3-diol Chemical compound OCC(CO)CO SFRDXVJWXWOTEW-UHFFFAOYSA-N 0.000 description 1
- IEQAICDLOKRSRL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-dodecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO IEQAICDLOKRSRL-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- WDBQJSCPCGTAFG-QHCPKHFHSA-N 4,4-difluoro-N-[(1S)-3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-pyridin-3-ylpropyl]cyclohexane-1-carboxamide Chemical compound FC1(CCC(CC1)C(=O)N[C@@H](CCN1CCC(CC1)N1C(=NN=C1C)C(C)C)C=1C=NC=CC=1)F WDBQJSCPCGTAFG-QHCPKHFHSA-N 0.000 description 1
- KFGOFTHODYBSGM-IJCBKZNRSA-N 6-Keto-prostaglandin F1a Chemical compound CCCCC[C@H](O)C=C[C@H]1[C@H](O)C[C@H](O)[C@@H]1CC(=O)CCCCC(O)=O KFGOFTHODYBSGM-IJCBKZNRSA-N 0.000 description 1
- KFGOFTHODYBSGM-UHFFFAOYSA-N 6-Oxoprostaglandin F1alpha Natural products CCCCCC(O)C=CC1C(O)CC(O)C1CC(=O)CCCCC(O)=O KFGOFTHODYBSGM-UHFFFAOYSA-N 0.000 description 1
- 206010000084 Abdominal pain lower Diseases 0.000 description 1
- PQSUYGKTWSAVDQ-ZVIOFETBSA-N Aldosterone Chemical compound C([C@@]1([C@@H](C(=O)CO)CC[C@H]1[C@@H]1CC2)C=O)[C@H](O)[C@@H]1[C@]1(C)C2=CC(=O)CC1 PQSUYGKTWSAVDQ-ZVIOFETBSA-N 0.000 description 1
- PQSUYGKTWSAVDQ-UHFFFAOYSA-N Aldosterone Natural products C1CC2C3CCC(C(=O)CO)C3(C=O)CC(O)C2C2(C)C1=CC(=O)CC2 PQSUYGKTWSAVDQ-UHFFFAOYSA-N 0.000 description 1
- 108010064733 Angiotensins Proteins 0.000 description 1
- 102000015427 Angiotensins Human genes 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 206010003658 Atrial Fibrillation Diseases 0.000 description 1
- NOWKCMXCCJGMRR-UHFFFAOYSA-N Aziridine Chemical compound C1CN1 NOWKCMXCCJGMRR-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108010075254 C-Peptide Proteins 0.000 description 1
- 235000021318 Calcifediol Nutrition 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 101800001982 Cholecystokinin Proteins 0.000 description 1
- 102100025841 Cholecystokinin Human genes 0.000 description 1
- 102000011022 Chorionic Gonadotropin Human genes 0.000 description 1
- 108010062540 Chorionic Gonadotropin Proteins 0.000 description 1
- 102100021809 Chorionic somatomammotropin hormone 1 Human genes 0.000 description 1
- 244000060011 Cocos nucifera Species 0.000 description 1
- 235000013162 Cocos nucifera Nutrition 0.000 description 1
- 206010010305 Confusional state Diseases 0.000 description 1
- 208000002330 Congenital Heart Defects Diseases 0.000 description 1
- 101800000414 Corticotropin Proteins 0.000 description 1
- UIKROCXWUNQSPJ-UHFFFAOYSA-N Cotinine Natural products C1CC(=O)N(C)C1C1=CC=CN=C1 UIKROCXWUNQSPJ-UHFFFAOYSA-N 0.000 description 1
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 description 1
- 229930105110 Cyclosporin A Natural products 0.000 description 1
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 1
- 108010036949 Cyclosporine Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FMGSKLZLMKYGDP-UHFFFAOYSA-N Dehydroepiandrosterone Natural products C1C(O)CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CC=C21 FMGSKLZLMKYGDP-UHFFFAOYSA-N 0.000 description 1
- 208000020401 Depressive disease Diseases 0.000 description 1
- 241000208011 Digitalis Species 0.000 description 1
- WDJUZGPOPHTGOT-OAXVISGBSA-N Digitoxin Natural products O([C@H]1[C@@H](C)O[C@@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@@](C)([C@H](C6=CC(=O)OC6)CC5)CC4)CC3)CC2)C[C@H]1O)[C@H]1O[C@@H](C)[C@H](O[C@H]2O[C@@H](C)[C@@H](O)[C@@H](O)C2)[C@@H](O)C1 WDJUZGPOPHTGOT-OAXVISGBSA-N 0.000 description 1
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000012286 ELISA Assay Methods 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 108090000394 Erythropoietin Proteins 0.000 description 1
- 102000003951 Erythropoietin Human genes 0.000 description 1
- DNXHEGUUPJUMQT-CBZIJGRNSA-N Estrone Chemical compound OC1=CC=C2[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 DNXHEGUUPJUMQT-CBZIJGRNSA-N 0.000 description 1
- 208000010201 Exanthema Diseases 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 102000012673 Follicle Stimulating Hormone Human genes 0.000 description 1
- 108010079345 Follicle Stimulating Hormone Proteins 0.000 description 1
- 101800002068 Galanin Proteins 0.000 description 1
- 102000019432 Galanin Human genes 0.000 description 1
- 102400000921 Gastrin Human genes 0.000 description 1
- 108010052343 Gastrins Proteins 0.000 description 1
- 108010007979 Glycocholic Acid Proteins 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 239000007821 HATU Substances 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- DOMWKUIIPQCAJU-LJHIYBGHSA-N Hydroxyprogesterone caproate Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(C)=O)(OC(=O)CCCCC)[C@@]1(C)CC2 DOMWKUIIPQCAJU-LJHIYBGHSA-N 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- 241001468196 Leuconostoc pseudomesenteroides Species 0.000 description 1
- 102000009151 Luteinizing Hormone Human genes 0.000 description 1
- 108010073521 Luteinizing Hormone Proteins 0.000 description 1
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 101710151321 Melanostatin Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- LFZAGIJXANFPFN-UHFFFAOYSA-N N-[3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-thiophen-2-ylpropyl]acetamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CCC(C=1SC=CC=1)NC(C)=O)C LFZAGIJXANFPFN-UHFFFAOYSA-N 0.000 description 1
- RFDAIACWWDREDC-UHFFFAOYSA-N Na salt-Glycocholic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCC(O)=O)C)C1(C)C(O)C2 RFDAIACWWDREDC-UHFFFAOYSA-N 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 101800003845 Neuropeptide Y Proteins 0.000 description 1
- 101800001814 Neurotensin Proteins 0.000 description 1
- 102000050267 Neurotensin Human genes 0.000 description 1
- 102000004067 Osteocalcin Human genes 0.000 description 1
- 108090000573 Osteocalcin Proteins 0.000 description 1
- 108090000445 Parathyroid hormone Proteins 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- CXOFVDLJLONNDW-UHFFFAOYSA-N Phenytoin Chemical compound N1C(=O)NC(=O)C1(C=1C=CC=CC=1)C1=CC=CC=C1 CXOFVDLJLONNDW-UHFFFAOYSA-N 0.000 description 1
- 108010003044 Placental Lactogen Proteins 0.000 description 1
- 239000000381 Placental Lactogen Substances 0.000 description 1
- 239000004353 Polyethylene glycol 8000 Substances 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 102100027467 Pro-opiomelanocortin Human genes 0.000 description 1
- 108010048233 Procalcitonin Proteins 0.000 description 1
- 108010076181 Proinsulin Proteins 0.000 description 1
- 102000003946 Prolactin Human genes 0.000 description 1
- 108010057464 Prolactin Proteins 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
- 241000700159 Rattus Species 0.000 description 1
- 102100028255 Renin Human genes 0.000 description 1
- 108090000783 Renin Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010086019 Secretin Proteins 0.000 description 1
- 102100037505 Secretin Human genes 0.000 description 1
- 206010040738 Sinus arrest Diseases 0.000 description 1
- 208000032140 Sleepiness Diseases 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 108010056088 Somatostatin Proteins 0.000 description 1
- 102000005157 Somatostatin Human genes 0.000 description 1
- 102100038803 Somatotropin Human genes 0.000 description 1
- 206010041349 Somnolence Diseases 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 208000003734 Supraventricular Tachycardia Diseases 0.000 description 1
- 206010049447 Tachyarrhythmia Diseases 0.000 description 1
- 208000001871 Tachycardia Diseases 0.000 description 1
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 description 1
- XNRNNGPBEPRNAR-UHFFFAOYSA-N Thromboxane B2 Natural products CCCCCC(O)C=CC1OC(O)CC(O)C1CC=CCCCC(O)=O XNRNNGPBEPRNAR-UHFFFAOYSA-N 0.000 description 1
- 102000011923 Thyrotropin Human genes 0.000 description 1
- 108010061174 Thyrotropin Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 229920004896 Triton X-405 Polymers 0.000 description 1
- 208000024780 Urticaria Diseases 0.000 description 1
- 108010004977 Vasopressins Proteins 0.000 description 1
- 102000002852 Vasopressins Human genes 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229960002478 aldosterone Drugs 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 229960005260 amiodarone Drugs 0.000 description 1
- IYIKLHRQXLHMJQ-UHFFFAOYSA-N amiodarone Chemical compound CCCCC=1OC2=CC=CC=C2C=1C(=O)C1=CC(I)=C(OCCN(CC)CC)C(I)=C1 IYIKLHRQXLHMJQ-UHFFFAOYSA-N 0.000 description 1
- AEMFNILZOJDQLW-QAGGRKNESA-N androst-4-ene-3,17-dione Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 AEMFNILZOJDQLW-QAGGRKNESA-N 0.000 description 1
- 229960003473 androstanolone Drugs 0.000 description 1
- 229960005471 androstenedione Drugs 0.000 description 1
- AEMFNILZOJDQLW-UHFFFAOYSA-N androstenedione Natural products O=C1CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 AEMFNILZOJDQLW-UHFFFAOYSA-N 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- KBZOIRJILGZLEJ-LGYYRGKSSA-N argipressin Chemical compound C([C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@@H](C(N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N1)=O)N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(N)=O)C1=CC=CC=C1 KBZOIRJILGZLEJ-LGYYRGKSSA-N 0.000 description 1
- 206010003119 arrhythmia Diseases 0.000 description 1
- 206010003668 atrial tachycardia Diseases 0.000 description 1
- 239000007982 barbital buffer Substances 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 150000003943 catecholamines Chemical class 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- AOXOCDRNSPFDPE-UKEONUMOSA-N chembl413654 Chemical compound C([C@H](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](C)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@@H](N)CCC(O)=O)C1=CC=C(O)C=C1 AOXOCDRNSPFDPE-UKEONUMOSA-N 0.000 description 1
- 229940107137 cholecystokinin Drugs 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 229960001265 ciclosporin Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 208000028831 congenital heart disease Diseases 0.000 description 1
- IDLFZVILOHSSID-OVLDLUHVSA-N corticotropin Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](N)CO)C1=CC=C(O)C=C1 IDLFZVILOHSSID-OVLDLUHVSA-N 0.000 description 1
- 229960000258 corticotropin Drugs 0.000 description 1
- 229950006073 cotinine Drugs 0.000 description 1
- ZOOGRGPOEVQQDX-KHLHZJAASA-N cyclic guanosine monophosphate Chemical compound C([C@H]1O2)O[P@](O)(=O)O[C@@H]1[C@H](O)[C@H]2N1C(N=C(NC2=O)N)=C2N=C1 ZOOGRGPOEVQQDX-KHLHZJAASA-N 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- FMGSKLZLMKYGDP-USOAJAOKSA-N dehydroepiandrosterone Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC=C21 FMGSKLZLMKYGDP-USOAJAOKSA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 1
- 229960000648 digitoxin Drugs 0.000 description 1
- WDJUZGPOPHTGOT-XUDUSOBPSA-N digitoxin Chemical compound C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)CC5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O WDJUZGPOPHTGOT-XUDUSOBPSA-N 0.000 description 1
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 description 1
- 239000002934 diuretic Substances 0.000 description 1
- AFOSIXZFDONLBT-UHFFFAOYSA-N divinyl sulfone Chemical compound C=CS(=O)(=O)C=C AFOSIXZFDONLBT-UHFFFAOYSA-N 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 229960005139 epinephrine Drugs 0.000 description 1
- 229960001123 epoprostenol Drugs 0.000 description 1
- KAQKFAOMNZTLHT-VVUHWYTRSA-N epoprostenol Chemical compound O1C(=CCCCC(O)=O)C[C@@H]2[C@@H](/C=C/[C@@H](O)CCCCC)[C@H](O)C[C@@H]21 KAQKFAOMNZTLHT-VVUHWYTRSA-N 0.000 description 1
- 229940105423 erythropoietin Drugs 0.000 description 1
- 229960005309 estradiol Drugs 0.000 description 1
- 229930182833 estradiol Natural products 0.000 description 1
- 229960001348 estriol Drugs 0.000 description 1
- PROQIPRRNZUXQM-ZXXIGWHRSA-N estriol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H]([C@H](O)C4)O)[C@@H]4[C@@H]3CCC2=C1 PROQIPRRNZUXQM-ZXXIGWHRSA-N 0.000 description 1
- 229960003399 estrone Drugs 0.000 description 1
- 201000005884 exanthem Diseases 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
- 238000001914 filtration Methods 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 229940028334 follicle stimulating hormone Drugs 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- RFDAIACWWDREDC-FRVQLJSFSA-N glycocholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 RFDAIACWWDREDC-FRVQLJSFSA-N 0.000 description 1
- 229940099347 glycocholic acid Drugs 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 208000018578 heart valve disease Diseases 0.000 description 1
- 229960001340 histamine Drugs 0.000 description 1
- 229940084986 human chorionic gonadotropin Drugs 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 229950000801 hydroxyprogesterone caproate Drugs 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 208000021803 junctional tachycardia Diseases 0.000 description 1
- GUWHRJQTTVADPB-UHFFFAOYSA-N lithium azide Chemical compound [Li+].[N-]=[N+]=[N-] GUWHRJQTTVADPB-UHFFFAOYSA-N 0.000 description 1
- 229940040129 luteinizing hormone Drugs 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 1
- 229960002510 mandelic acid Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 201000003102 mental depression Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- SLZIZIJTGAYEKK-CIJSCKBQSA-N molport-023-220-247 Chemical compound C([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)CN)[C@@H](C)O)C1=CNC=N1 SLZIZIJTGAYEKK-CIJSCKBQSA-N 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- 230000001452 natriuretic effect Effects 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- PCJGZPGTCUMMOT-ISULXFBGSA-N neurotensin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1NC(=O)CC1)C1=CC=C(O)C=C1 PCJGZPGTCUMMOT-ISULXFBGSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229960002748 norepinephrine Drugs 0.000 description 1
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- URPYMXQQVHTUDU-OFGSCBOVSA-N nucleopeptide y Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 URPYMXQQVHTUDU-OFGSCBOVSA-N 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 239000000199 parathyroid hormone Substances 0.000 description 1
- 230000001314 paroxysmal effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 229960002036 phenytoin Drugs 0.000 description 1
- 229920000768 polyamine Polymers 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 235000019446 polyethylene glycol 8000 Nutrition 0.000 description 1
- 229940085678 polyethylene glycol 8000 Drugs 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000009090 positive inotropic effect Effects 0.000 description 1
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 description 1
- 229960002847 prasterone Drugs 0.000 description 1
- 230000001536 pro-arrhythmogenic effect Effects 0.000 description 1
- CWCXERYKLSEGEZ-KDKHKZEGSA-N procalcitonin Chemical compound C([C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCSC)NC(=O)[C@H]1NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@@H](N)CSSC1)[C@@H](C)O)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 CWCXERYKLSEGEZ-KDKHKZEGSA-N 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 229960003387 progesterone Drugs 0.000 description 1
- 239000000186 progesterone Substances 0.000 description 1
- 229940097325 prolactin Drugs 0.000 description 1
- 150000003180 prostaglandins Chemical class 0.000 description 1
- 229940001470 psychoactive drug Drugs 0.000 description 1
- 239000004089 psychotropic agent Substances 0.000 description 1
- 206010037844 rash Diseases 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229960002101 secretin Drugs 0.000 description 1
- OWMZNFCDEHGFEP-NFBCVYDUSA-N secretin human Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(N)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)C1=CC=CC=C1 OWMZNFCDEHGFEP-NFBCVYDUSA-N 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 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 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 1
- 229960000553 somatostatin Drugs 0.000 description 1
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- ADNPLDHMAVUMIW-CUZNLEPHSA-N substance P Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CCCN=C(N)N)C1=CC=CC=C1 ADNPLDHMAVUMIW-CUZNLEPHSA-N 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 1
- 229960001967 tacrolimus Drugs 0.000 description 1
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 229960000278 theophylline Drugs 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- DSNBHJFQCNUKMA-SCKDECHMSA-N thromboxane A2 Chemical compound OC(=O)CCC\C=C/C[C@@H]1[C@@H](/C=C/[C@@H](O)CCCCC)O[C@@H]2O[C@H]1C2 DSNBHJFQCNUKMA-SCKDECHMSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 1
- 229960000281 trometamol Drugs 0.000 description 1
- 230000002227 vasoactive effect Effects 0.000 description 1
- 229960003726 vasopressin Drugs 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
- 208000003663 ventricular fibrillation Diseases 0.000 description 1
- 206010047302 ventricular tachycardia Diseases 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The present application relates to a process for the preparation of conjugates. Specifically, the mutant glucose 6-phosphate dehydrogenase of the present application comprises one mutation or a combination thereof selected from the group consisting of: d306C, D375C, G426C. The detection kit prepared by using the glucose 6-phosphate dehydrogenase mutant has the advantages of strong specificity, high sensitivity, convenient operation, short detection time, accurate quantification and suitability for high-throughput detection.
Description
The application relates to a patent application 202010004879.2 '6-phosphoglucose dehydrogenase mutant and application thereof in preparing digoxin detection reagent', which are filed 1 month and 3 days in 2020.
Technical Field
The application relates to the field of biological detection, in particular to mutant enzyme glucose 6-phosphate dehydrogenase (G6 PDH for short) and application thereof in a digoxin detection kit.
Background
Hapten, some small molecule substances (molecular weight less than 4000 Da) alone are not able to induce an immune response, i.e. are not immunogenic, but are immunogenic when crosslinked or conjugated to a carrier such as a macromolecular protein or non-antigenic polylysine, inducing an immune response. These small molecule substances can bind to the response effect products, are antigenic, are only immunoreactive, are not immunogenic, and are also called incomplete antigens.
Hapten can bind to the corresponding antibody to generate antigen-antibody reaction, and antigen which can not independently excite human or animal body to generate antibody can not be generated. It is only immunoreactive, not immunogenic, also known as incomplete antigen. Most polysaccharides, lipids, hormones and small molecule drugs belong to the hapten group. If hapten is chemically bound to a protein molecule (carrier), new immunogenicity is obtained and the animal is stimulated to produce the corresponding antibody. Hapten, once bound to a protein, constitutes an antigenic cluster of the protein. Some chemically active substances (such as penicillin, sulfonamides, etc.) which have a smaller molecular weight than the general hapten but a specific structure are called simple haptens.
Small molecule antigens or haptens, which lack two or more sites available for sandwich methods, cannot be assayed by the double antibody sandwich method, and are often in competition mode. The principle is that the antigen in the specimen and a certain amount of enzyme-labeled antigen compete for binding with the solid phase antibody. The more the antigen content in the specimen, the less the enzyme-labeled antigen is bound on the solid phase, and the lighter the color development. ELISA assay for small molecule hormones, drugs and the like is commonly used.
Digoxin (Digoxin) has the structural formula shown below:
digoxin is a cardiac glycoside drug which has positive inotropic effect on heart, slows down heart rate and inhibits heart conduction, and is used for treating acute and chronic cardiac insufficiency such as hypertension, valvular heart disease, congenital heart disease and the like. Is particularly suitable for cardiac insufficiency associated with atrial fibrillation with rapid ventricular rate.
Common adverse reactions include: proarrhythmic effects, nausea, vomiting, lower abdominal pain, abnormal weakness, weakness; rare reactions include: vision blur, central nervous system response (e.g., mental depression or confusion); rare reactions include: somnolence, headache, rash, hives (allergic reactions). Among the toxic manifestations of digitalis, tachyarrhythmia is most important, and secondarily atrioventricular block, paroxysmal or accelerating junctional tachycardia, paroxysmal atrial tachycardia with atrioventricular block, ventricular tachycardia, sinus arrest, ventricular fibrillation, etc.
Therefore, attention is paid to monitoring adverse drug reactions. And because of differences of individual metabolisms of the drugs, the drug administration method should be combined with blood concentration monitoring during clinical use to formulate a reasonable dosing scheme so as to avoid adverse reactions as much as possible.
The currently known digoxin detection methods mainly comprise the following steps: high Performance Liquid Chromatography (HPLC), chemiluminescence immunity, enzyme-linked immunosorbent assay (ELISA), homogeneous enzyme immunoassay, latex agglutination turbidimetry, etc. The HPLC method requires complex sample pretreatment, and has the advantages of long operation period and high cost; the luminous immunoassay method has the disadvantages of high reagent cost, inapplicability to detection of conventional therapeutic drugs and inapplicability to large-scale popularization. The existing homogeneous enzyme immunoassay and latex agglutination turbidimetry are often limited in application due to complex preparation process and large batch-to-batch difference.
The prior art CN108593905a describes a digoxin assay kit and a method for preparing the same. However, the prior art methods rely on activation of the reactive groups carried by the small molecule drug (digoxin) itself before reaction with the enzyme. Such coupling methods can occur when there are multiple digoxigenin linked to the same glucose hexaphosphate dehydrogenase, and it is difficult to ensure consistency of the coupling sites, and it is difficult to ensure orientation between the small molecule drug and the enzyme 1:1, resulting in large batch-to-batch variation.
Disclosure of Invention
In view of the needs in the art, the present application provides a novel glucose 6-phosphate dehydrogenase mutant and its use in preparing digoxin assay kits.
According to some embodiments, a glucose 6-phosphate dehydrogenase mutant is provided. Unlike the mutants of glucose 6 phosphate dehydrogenase of the prior published patent US006090567A (Homogeneous immunoassays using mutant glucose-6-phosphate dehydrogenases), the glucose 6-phosphate dehydrogenase mutant of the present application comprises a mutation selected from the group consisting of: d306C, D375C, G426C.
According to some embodiments, there is provided a glucose 6-phosphate dehydrogenase mutant, the glucose 6-phosphate dehydrogenase mutant being represented by a sequence selected from the group consisting of: SEQ ID No.2, SEQ ID No.3, SEQ ID No.4.
According to some embodiments, a polynucleotide encoding a glucose 6-phosphate dehydrogenase mutant of the present application is provided.
According to some embodiments, there is provided an expression vector comprising a polynucleotide of the application.
According to some embodiments, there is provided a host cell comprising an expression vector of the application. The host cell may be prokaryotic (e.g., bacteria) or eukaryotic (e.g., yeast).
According to some embodiments, there is provided a conjugate which is a glucose 6-phosphate dehydrogenase mutant of the present application and a hapten in a molar ratio of 1: and n is coupled.
In some embodiments, n is 1 to 50, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50.
In some specific embodiments, the molar ratio of glucose 6-phosphate dehydrogenase mutants of the present application to hapten is preferably 1:1.
in some specific embodiments, the hapten has a molecular weight of 100Da to 4000Da, for example: 100. 150, 200, 250, 300, 350, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 520, 550, 570, 600, 620, 650, 700, 750, 800, 850, 900, 950, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000.
According to the present application, the skilled person will understand that "hapten" also includes the form of its derivative. In order to facilitate the coupling with glucose-6-phosphate dehydrogenase, haptens (e.g., digoxin) that do not themselves carry a coupling group (e.g., a group that reacts with a thiol group) may be engineered to carry a linker for covalent binding to the thiol group. Thus, in the present application, hapten derivatives refer to haptens engineered to bear a thiol-reactive group.
The hapten is selected from the group consisting of: small molecule drugs (e.g., antibiotics, psychotropic drugs), hormones, metabolites, sugars, lipids, and amino acids.
Hapten such as, but not limited to: theophylline, phenytoin, vitamin D, 25 hydroxy vitamin D, 1, 25 dihydroxyvitamin D, folic acid, cardiac glycoside (including digoxin, digitoxin), zymophenolic acid, lei Paming, cyclosporin A, amiodarone, methotrexate, tacrolimus, serum amino acids, bile acids, glycocholic acid, phenylalanine, ethanol, the product of the uronictin metabolite, cotinine, uromorphine, uromonohydric phenol derivatives, neuropeptide tyrosine, plasma galanin, polyamines, histamine, thyroid stimulating hormone, prolactin, placental lactogen, growth hormone, follicle stimulating hormone, luteinizing hormone, adrenocorticotropin, antidiuretic hormone, calcitonin, procalcitonin, parathyroid hormone, thyroxine, triiodothyronine, anti-triiodothyronine, free thyroxine, free triiodothyronine, cortisol urine 17-hydroxycortic steroids, urine 17-ketosterols, dehydroepiandrosterone and sulfates, aldosterone, uronolamine mandelic acid, plasma renin, angiotensin, erythropoietin, testosterone, dihydrotestosterone, androstenedione, 17 alpha-hydroxyprogesterone, estrone, estriol, estradiol, progesterone, human chorionic gonadotropin, insulin, proinsulin, C peptide, gastrin, plasma prostaglandin, plasma 6-keto prostaglandin f1α, prostacyclin, epinephrine, catecholamine, norepinephrine, cholecystokinin, natriuretic acid adenosine cyclophosphate, cyclic guanosine monophosphate, vasoactive peptides, somatostatin, secretin, P-substance, neurotensin, thromboxane A2, thromboxane B2, 5 hydroxytryptamine, neuropeptide Y, osteocalcin.
In a specific embodiment, the hapten is digoxin or a derivative thereof.
In a specific embodiment, the hapten is a digoxin derivative having a sulfhydryl reactive group such as, for example, a maleimide, bromoacetyl, vinyl sulfone, or aziridine.
In a specific embodiment, the hapten is a digoxin derivative, as shown in formula I:
wherein ,
in some embodiments, m is an integer from 1 to 10, preferably an integer from 1 to 5, such as 1, 2, 3, 4, 5.
In some specific embodiments, the digoxin derivative has a structure represented by formula I-1:
wherein ,
according to some embodiments, there is provided an agent comprising a conjugate of the application.
According to some embodiments, there is provided the use of a glucose 6-phosphate dehydrogenase mutant of the application in the preparation of a digoxin assay reagent.
According to some embodiments, there is provided the use of a conjugate of the application in the preparation of a digoxin assay reagent.
In specific embodiments, the detection reagent is selected from the group consisting of: ELISA detection reagent, chemiluminescent detection reagent, homogeneous ELISA detection reagent and latex enhanced turbidimetry detection reagent.
In a specific embodiment, the detection reagent is preferably a reagent for competition-based detection.
According to some embodiments, there is provided the use of a conjugate of the application in the preparation of a digoxin assay device.
In particular embodiments, the detection device may be prepared in the form of a well plate (e.g., 96-well plate), such as a plate coated with reagents according to the application.
In a specific embodiment, the detection device may be prepared in the form of particles (e.g. latex, magnetic beads), such as particles coated with the reagent according to the application.
According to some embodiments, there is provided a digoxin assay kit comprising:
-a first reagent comprising a substrate, a buffer and a digoxin antibody; the substrate is a substrate for glucose-6-phosphate dehydrogenase;
-a second agent comprising a conjugate of the application and a buffer;
-optionally, a calibrator comprising 10mM to 500mM buffer, 0ng/ml to 5ng/ml digoxin (e.g. 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5ng/ml or any value in between); and
-optionally, a quality control comprising 10mM to 500mM buffer, 0.7ng/ml to 4.5ng/ml (e.g. 0.7, 0.8, 1.5, 1.8, 2, 3, 4, 4.5ng/ml or any value in between) digoxin.
According to one embodiment, there is provided a digoxin assay kit comprising:
a first reagent comprising:
10mM to 500mM buffer,
5mM to 50mM substrate,
0.01 to 10. Mu.g/ml of digoxin antibody (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.5, 2, 3, 4, 5. Mu.g/ml),
0.1g/L to 5g/L of stabilizer,
0.1g/L to 5g/L of surfactant,
0.1g/L to 5g/L preservative;
a second reagent comprising:
10mM to 500mM buffer,
0.01. Mu.g/ml to 10. Mu.g/ml of the conjugate according to the application (0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0. Mu.g/ml),
0.1g/L to 5g/L of stabilizer,
0.1g/L to 5g/L of surfactant,
0.1g/L to 5g/L preservative.
In some embodiments, the buffer is selected from one or a combination of the following: TAPS, tromethamine buffer, phosphate buffer, tris-HCl buffer, citric acid-sodium citrate buffer, barbital buffer, glycine buffer, borate buffer, and trimethylol methane buffer; preferably, a phosphate buffer; the concentration of the buffer is 10mmol/L to 500mmol/L, preferably 50 to 100mM; the pH of the buffer is 7 to 8.
In some embodiments, the stabilizer is selected from one or a combination of the following: bovine serum albumin, trehalose, glycerol, sucrose, mannitol, glycine, arginine, polyethylene glycol 6000, polyethylene glycol 8000; bovine serum albumin is preferred.
In some embodiments, the surfactant is selected from one or a combination of the following: brij23, brij35, triton X-100, triton X-405, tween20, tween30, tween80, coconut fatty acid diethanolamide, AEO7, preferably Tween20.
In some embodiments, the preservative is selected from one or a combination of the following: azide, MIT, biological preservative PC (such as PC-300), merthiolate; the azide is selected from: sodium azide, lithium azide, PC-300.
In some embodiments, the substrate comprises: glucose-6-phosphate, beta-nicotinamide adenine dinucleotide.
In some specific embodiments, the digoxin antibody is derived from: mice, rats, cats, dogs, primates, cows, horses, sheep, camelids, birds, humans.
In some specific embodiments, the digoxin antibody is selected from the group consisting of: monoclonal antibodies, polyclonal antibodies, recombinant antibodies, chimeric antibodies, and antigen binding fragments.
According to some embodiments, there is provided a method of preparing a conjugate comprising the steps of:
1) Providing a digoxin derivative according to the present application, particularly in an aprotic solvent (e.g., without limitation, acetonitrile, dimethylformamide, dimethylsulfoxide);
2) Providing a glucose 6-phosphate dehydrogenase mutant, preferably in a buffer (which provides a reaction environment such as, but not limited to PBS, tris, TAPS, TAPSO, said buffer having a pH of 6.0 to 8.0);
3) At 18 ℃ to 28 ℃, the glucose 6-phosphate dehydrogenase mutant and the digoxin derivative are prepared according to the digoxin derivative: enzyme = 500:1 to 1:500 (preferably 50:1 to 1:50) for 1 to 4 hours (1, 1.5, 2, 2.5, 3, 3.5, 4 hours, or any value in between, preferably 2 hours to 3 hours) such that the digoxin derivative and the glucose 6-phosphate dehydrogenase mutant are coupled to give the seed conjugate;
4) The seed conjugate is optionally purified, e.g., desalted, etc., as desired.
In some embodiments, the hapten and enzyme are contacted in a molar ratio of 1: n, where n is 1 to 500, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 100, 200, 300, 400, 500, and ranges between any of the foregoing values thereof.
In some specific embodiments, steps 1) and 2) may be interchanged or in parallel.
In some specific embodiments, the glucose 6-phosphate dehydrogenase comprises one or more free sulfhydryl groups prior to coupling, thereby allowing for a directed reaction with digoxin.
Wild-type glucose 6-phosphate dehydrogenase does not contain a free thiol group, and thus in some specific embodiments, the glucose 6-phosphate dehydrogenase is genetically engineered to have an amino acid mutation at a particular site (306, 375, or 426) to a cysteine, thereby carrying a free thiol group.
Drawings
FIG. 1G 6PDH (wild type) amino acid sequence (SEQ ID No. 1); is derived from Leuconostoc pseudomesenteroides Leuconostoc pseudomesenteroides.
FIG. 2G 6PDH (D306C) amino acid sequence (SEQ ID No. 2).
FIG. 3G 6PDH (D375C) amino acid sequence (SEQ ID No. 3).
FIG. 4G 6PDH (G426C) amino acid sequence (SEQ ID No. 4).
Detailed Description
Examples
EXAMPLE 1 Synthesis of digoxin derivatives
1. Synthesis of Compound 2
1.0g of digoxin was dissolved in 95% ethanol (80 ml), followed by adding a solution of periodic acid (1.0 g) in water (10 ml) thereto, and stirring at room temperature (18-28 ℃ C.) for 1 hour. The residue was removed by filtration, the solvent was removed under reduced pressure, and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure to give compound 2 (white solid, 0.99g, 99%).
2. Synthesis of Compound 4
Compound 2 (900 mg,1.16 mmol) was dissolved in 10ml of dry methanol, and compound 3 (318 mg,1.0 mmol) was added to the reaction system and stirred at room temperature for 5 minutes. Sodium cyanoborohydride (146 mg,2.32 mmol) was added and stirred at room temperature for about 12 hours. The solvent was removed under reduced pressure and purified by direct column chromatography to give compound 4 (white solid, 585mg, 66%).
3. Synthesis of Compound 5
Compound 4 was dissolved in 15ml of dichloromethane and stirred at room temperature under nitrogen for 30 minutes, then 10ml of piperidine was added and stirred at room temperature for 2 hours. The solution was removed under reduced pressure, and purified by column chromatography to give compound 5 (460 mg, 78%).
4. Synthesis of digoxin derivatives
Compound 5 (88 mg,0.11 mmol) and compound 6 (17 mg,0.11 mmol) were dissolved in DCM (8 mL), triethylamine (33 mg,0.33 mmol) was added dropwise thereto, then HATU (50 mg,0.13 mmol) was added and stirred at room temperature for 2h to give the digoxin derivative (white solid, 50mg, 45%).
5. The structure of the digoxin derivative is correct through mass spectrum and nuclear magnetism identification.
This example allows digoxin to carry a group that can bind to enzymes.
EXAMPLE 2 coupling of digoxin derivatives to G6PDH molecules
1. The coupling method of the application
The G6 PDH-digoxin conjugates according to the application were coupled as follows: thiol-reactive groups (such as but not limited to maleimide groups) on digoxin derivative molecules are covalently bound to thiol groups on G6PDH molecules.
1. The digoxin derivative prepared in example 1 was dissolved in N, N-dimethylformamide (10 mg/ml);
g6pdh solution: g6pdh (e.g. mutant of the application) was dissolved in PB 100mmol, naCl 100mmol, ph=8.0;
3. 200 mu l G of 6PDH solution was added to 750. Mu.l buffer (0.05M Na 2 HPO 4 、150mM NaCl、10mM EDTA、0.1% NaN 3 Ph=7.2); then 50. Mu.l of an N, N-dimethylformamide solution of a digoxin derivative was added thereto;
4. the mixed solution is fully vibrated for 2-3 hours at room temperature (18-28 ℃), desalted and protein peaks are collected, and the obtained product is the G6 PDH-digoxin conjugate.
2. Control coupling method (refer to CN 108593905A)
Accurately weighing 100-300mg digoxin, and dissolving with 5-15mL absolute ethyl alcohol;
dropwise adding 5-15mL of 10-200mM sodium periodate into the solution, slightly oscillating, and stirring at room temperature for reaction for 0.5-2 hours;
dripping 0.5-2M glycol 0.5-1mL, stirring at room temperature, and reacting for 5-10 min;
dropwise adding the reaction mixture into 5-15mL of 2-3% G6PDH solution under stirring, regulating the pH of the solution to 9.0-9.5, continuously stirring and reacting for 0.5-2 hours, and stabilizing the pH of the solution;
adding 100-200mg of sodium tetrahydroborate, and stirring and reducing for 12-24 hours;
purifying by a G-25 gel chromatographic column to obtain the G6 PDH-digoxin conjugate.
EXAMPLE 3 preparation of the kit
The following kit for detecting digoxin was prepared, comprising:
reagent R1 comprising:
TAPS buffer 100mM, pH 7.0
15mM glucose 6-phosphate
15mM beta-nicotinamide adenine dinucleotide
0.5 μg/ml digoxin antibody (commercial antibody, without special restrictions)
1g/L bovine serum albumin
1g/L Brij
1g/L sodium azide;
reagent R2, comprising:
phosphate buffer 200mM, pH8.0
0.1 μg/ml G6 PDH-digoxin conjugate
100mM NaCl
1g/L bovine serum albumin
1g/L Brij
1g/L sodium azide;
calibration material: 20mM HEPES buffer, 0ng/ml, 0.5ng/ml, 1ng/ml, 2ng/ml, 3ng/ml, 5ng/ml digoxin (or added as needed);
quality control product: 20mM HEPES buffer, 0.6-0.8ng/ml, 1.6-2.0ng/ml, 3.8-4.2ng/ml digoxin (or added as needed).
The reagent (optionally containing quality control materials and calibrator) is assembled into the digoxin homogeneous enzyme immunoassay kit.
Test case
Principle of homogeneous enzyme immunoassay: in a liquid homogeneous reaction system, an enzyme-labeled antigen (such as G6 PDH-digoxin) competes with a non-labeled antigen (digoxin) for binding with a quantitative antibody (digoxin antibody), and when the more the antibody is bound with the non-labeled antigen, the more activity the enzyme-labeled antigen releases, and the more NAD+ which is a substrate for enzyme catalysis generates NADH.
Detecting the absorbance change of NADH at 340nm wavelength to calculate the content of digoxin in the liquid.
TABLE 1 full automatic Biochemical instrument parameters
Test example 1. Performance of the kit of the application
1. Calibration experiment
TABLE 2 Digoxin detection kit scaled absorbance
2. Precision experiments
TABLE 3 Total imprecision
3. Repeatability of
TABLE 4 repeatability
4. Recovery test
TABLE 5 recovery data
5. Linear experiments
TABLE 6 linearity
Test example 2 on-board stability
The reagent (D375C mutant) has the advantages that the calibrated absorbance is reduced by less than 10% after the reagent is accelerated for 7 days at 37 ℃, and the calibrated absorbance is obviously reduced after the reagent is accelerated for 7 days at 37 ℃ in comparison with the reagent.
TABLE 7 accelerated stability at 37℃
Detection example 3 antibody inhibition Rate
1. Principle of detection of antibody inhibition
When the antibody is combined with the G6 PDH-digoxin conjugate, the activity of the G6PDH enzyme is influenced due to steric hindrance, so that the efficiency of catalyzing NAD to be converted into NADH is reduced, and the difference between an added antibody and an experimental group without the added antibody is compared by detecting the change of the NADH amount, wherein the difference is expressed as the inhibition capability of the antibody on the G6 PDH.
2. Reaction system
TABLE 8 preparation of reagents for detection of antibody inhibition
3. Results
And comparing the absorbance measurement value of the G6 PDH-digoxin conjugate when the antibody is added with the antibody is not added, and obtaining the inhibition condition of the antibody on the G6 PDH.
Antibody inhibition = change in absorbance of G6 PDH-digoxin conjugate with antibody/change in absorbance of G6 PDH-digoxin without antibody x 100%.
Compared with the published mutation site (A45C), the mutant of the application has obvious improvement in enzyme activity retention, and can reach more than 40% (G426C: 40%; D375C: 49%), up to 62% (D306C). Published mutation sites (e.g., A45C, K55C) were prepared as G6 PDH-digoxin conjugates with only 33% and 40% inhibition by reference to the methods of the application.
While not being limited to a particular theory, it may be explained in part as: in comparison with the G6PDH mutant (A45C, K C) in the prior art, the mutation site (i.e. the site for introducing free sulfhydryl) in the enzyme mutant is the coupling site with hapten (such as hormone, small molecule drug, etc.). When hapten is combined with hapten specific antibody at this position, the steric hindrance formed has the greatest effect on the activity of G6PDH enzyme, and after mutation is introduced, the steric folding of the molecule cannot be substantially influenced. Therefore, the position of this mutation site is very important, and it is necessary to combine the activity of the G6PDH enzyme, the spatial folding of the coupling molecule, and the sufficient exposure of the hapten epitope.
The mutant of the enzyme has obvious improvement on the inhibition rate of the antibody. After the conjugate of the enzyme mutant and the digoxin is prepared into the kit, the reagent has obvious performance improvement in the aspects of the inter-batch variation coefficient, linearity, repeatability, stability and the like.
Claims (2)
1. A method of preparing a conjugate comprising the steps of:
1) Providing a digoxin derivative;
2) Providing a glucose 6-phosphate dehydrogenase mutant;
3) The glucose 6-phosphate dehydrogenase mutant is coupled with the digoxin derivative;
the glucose 6-phosphate dehydrogenase mutant comprises a D306C mutation compared to the wild-type glucose 6-phosphate dehydrogenase; the glucose 6-phosphate dehydrogenase mutant is shown in SEQ ID No. 2;
the digoxin derivative is represented by formula I:
wherein ,
m is an integer from 1 to 10, preferably an integer from 1 to 5.
2. The method according to claim 1, comprising the steps of:
1) Providing the digoxin derivative, preferably in an aprotic solvent;
2) Providing the glucose 6-phosphate dehydrogenase mutant, preferably providing the glucose 6-phosphate dehydrogenase mutant in a buffer;
3) Contacting the digoxin derivative and the mutant glucose 6-phosphate dehydrogenase at 18 ℃ to 28 ℃ for 1 hour to 4 hours, preferably 2 hours to 3 hours, such that the digoxin derivative and the mutant glucose 6-phosphate dehydrogenase are coupled to obtain the conjugate;
4) Optionally, purifying, preferably desalting, the conjugate;
step 1) and step 2) can be interchanged or parallel;
the buffer is selected from one of the following: PBS, tris, TAPS, TAPSO the number of the individual pieces of the plastic,
the buffer pH is 6.0 to 8.0;
the aprotic solvent is selected from one or a combination of the following: acetonitrile, dimethylformamide, dimethyl sulfoxide;
preferably, prior to step 3), the glucose 6-phosphate dehydrogenase mutant has a free thiol group at position 306.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2019100177644 | 2019-01-09 | ||
CN201910017764 | 2019-01-09 | ||
CN2019104231224 | 2019-05-21 | ||
CN201910423122.4A CN110174363A (en) | 2019-01-09 | 2019-05-21 | Glucose-6-phosphate dehydrogenase mutant and its purposes in preparation detection reagent |
CN202010004879.2A CN111487207B (en) | 2019-01-09 | 2020-01-03 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010004879.2A Division CN111487207B (en) | 2019-01-09 | 2020-01-03 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116840467A true CN116840467A (en) | 2023-10-03 |
Family
ID=67691613
Family Applications (52)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910423122.4A Pending CN110174363A (en) | 2019-01-09 | 2019-05-21 | Glucose-6-phosphate dehydrogenase mutant and its purposes in preparation detection reagent |
CN202310257027.8A Pending CN116144619A (en) | 2019-01-09 | 2019-12-26 | Theophylline detection kit |
CN201911365439.3A Active CN111239060B (en) | 2019-01-09 | 2019-12-26 | 6-phosphoglucose dehydrogenase mutant and application thereof in preparing theophylline detection reagent |
CN202310217235.5A Pending CN116359146A (en) | 2019-01-09 | 2019-12-26 | Method for preparing conjugate |
CN201911372535.0A Active CN112285038B (en) | 2019-01-09 | 2019-12-27 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digitoxin detection reagent |
CN201911372147.2A Active CN112285037B (en) | 2019-01-09 | 2019-12-27 | 6-phosphoglucose dehydrogenase mutant and application thereof in preparing detection reagent |
CN202310811210.8A Pending CN116626281A (en) | 2019-01-09 | 2019-12-27 | Digitoxin detection kit |
CN202310811498.9A Pending CN116698772A (en) | 2019-01-09 | 2019-12-27 | Method for preparing conjugate |
CN202310811212.7A Pending CN116735512A (en) | 2019-01-09 | 2019-12-27 | Use of conjugates in the preparation of detection reagents |
CN202211151405.6A Pending CN115791649A (en) | 2019-01-09 | 2019-12-27 | Glycocholic acid detection kit |
CN202211151264.8A Pending CN116008201A (en) | 2019-01-09 | 2019-12-27 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of detection reagent |
CN202211153004.4A Pending CN115808398A (en) | 2019-01-09 | 2019-12-27 | Method for preparing conjugate |
CN202310726493.6A Pending CN116773795A (en) | 2019-01-09 | 2019-12-31 | Preparation method of conjugate |
CN202310726498.9A Pending CN116559472A (en) | 2019-01-09 | 2019-12-31 | Cortisol detection kit |
CN202310364266.3A Pending CN116718764A (en) | 2019-01-09 | 2019-12-31 | Use of conjugate in preparation of phenytoin detection reagent |
CN202310724066.4A Pending CN116773827A (en) | 2019-01-09 | 2019-12-31 | Use of conjugates in the preparation of detection reagents |
CN202310365160.5A Pending CN116355873A (en) | 2019-01-09 | 2019-12-31 | Preparation method of conjugate |
CN202310364997.8A Pending CN116298257A (en) | 2019-01-09 | 2019-12-31 | Phenytoin detection kit |
CN201911404154.6A Active CN111504920B (en) | 2019-01-09 | 2019-12-31 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of cortisol detection reagent |
CN201911403882.5A Active CN111650135B (en) | 2019-01-09 | 2019-12-31 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of phenytoin detection reagent |
CN202010000321.7A Active CN111487206B (en) | 2019-01-09 | 2020-01-02 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of vancomycin detection reagent |
CN202311025752.9A Pending CN117074335A (en) | 2019-01-09 | 2020-01-02 | Use of conjugates in the preparation of detection reagents |
CN202311025762.2A Pending CN117054643A (en) | 2019-01-09 | 2020-01-02 | Vancomycin detection kit |
CN202311025756.7A Pending CN117030640A (en) | 2019-01-09 | 2020-01-02 | Method for preparing conjugate |
CN202310811497.4A Pending CN116840468A (en) | 2019-01-09 | 2020-01-03 | Use of conjugates in the preparation of detection reagents |
CN202310810479.4A Pending CN116819060A (en) | 2019-01-09 | 2020-01-03 | Digoxin detection kit |
CN202310810455.9A Pending CN116840467A (en) | 2019-01-09 | 2020-01-03 | Method for preparing conjugate |
CN202010004879.2A Active CN111487207B (en) | 2019-01-09 | 2020-01-03 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
CN202010009771.2A Active CN111504921B (en) | 2019-01-09 | 2020-01-06 | 6-glucose phosphate dehydrogenase mutant and application thereof in preparation of gentamicin detection reagent |
CN202310320137.4A Pending CN116148198A (en) | 2019-01-09 | 2020-01-06 | Preparation method of gentamicin detection reagent |
CN202310452740.8A Pending CN116430056A (en) | 2019-01-09 | 2020-01-06 | Method for preparing conjugate |
CN202310320729.6A Pending CN116297271A (en) | 2019-01-09 | 2020-01-06 | Use of conjugates in the preparation of kits |
CN202310452946.0A Pending CN116559425A (en) | 2019-01-09 | 2020-01-06 | Use of conjugates in the preparation of detection reagents |
CN202010009570.2A Active CN111537451B (en) | 2019-01-09 | 2020-01-06 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of tacrolimus detection reagent |
CN202310453290.4A Pending CN116338215A (en) | 2019-01-09 | 2020-01-06 | Tacrolimus detection kit |
CN202310318754.0A Pending CN116124721A (en) | 2019-01-09 | 2020-01-06 | Gentamicin detection kit |
CN202310508418.2A Pending CN116381253A (en) | 2019-01-09 | 2020-01-07 | Method for preparing conjugate |
CN202010013644.XA Active CN111487208B (en) | 2019-01-09 | 2020-01-07 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of methotrexate detection reagent |
CN202310554774.8A Pending CN116679047A (en) | 2019-01-09 | 2020-01-07 | Method for preparing conjugate |
CN202010013174.7A Active CN111678874B (en) | 2019-01-09 | 2020-01-07 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of cyclosporine A detection reagent |
CN202310507880.0A Pending CN116754756A (en) | 2019-01-09 | 2020-01-07 | Methotrexate detection kit |
CN202310508217.2A Pending CN116298330A (en) | 2019-01-09 | 2020-01-07 | Use of conjugates in the preparation of detection reagents |
CN202310555230.3A Pending CN116718761A (en) | 2019-01-09 | 2020-01-07 | Cyclosporine A detection kit |
CN202310553479.0A Pending CN116699125A (en) | 2019-01-09 | 2020-01-07 | Use of conjugates in the preparation of detection reagents |
CN202310702870.2A Pending CN116840462A (en) | 2019-01-09 | 2020-01-08 | Method for preparing conjugate |
CN202010017376.9A Active CN111537452B (en) | 2019-01-09 | 2020-01-08 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of amikacin detection reagent |
CN202310725902.0A Pending CN116577494A (en) | 2019-01-09 | 2020-01-08 | Use of conjugates in the preparation of detection reagents |
CN202010016535.3A Active CN111693473B (en) | 2019-01-09 | 2020-01-08 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of rapamycin detection reagent |
CN202310726230.5A Pending CN116577495A (en) | 2019-01-09 | 2020-01-08 | Method for preparing conjugate |
CN202310726069.1A Pending CN116754761A (en) | 2019-01-09 | 2020-01-08 | Amikacin detection kit |
CN202310702858.1A Pending CN116859035A (en) | 2019-01-09 | 2020-01-08 | Use of conjugates in the preparation of detection reagents |
CN202310702860.9A Pending CN116699122A (en) | 2019-01-09 | 2020-01-08 | Rapamycin detection kit |
Family Applications Before (26)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910423122.4A Pending CN110174363A (en) | 2019-01-09 | 2019-05-21 | Glucose-6-phosphate dehydrogenase mutant and its purposes in preparation detection reagent |
CN202310257027.8A Pending CN116144619A (en) | 2019-01-09 | 2019-12-26 | Theophylline detection kit |
CN201911365439.3A Active CN111239060B (en) | 2019-01-09 | 2019-12-26 | 6-phosphoglucose dehydrogenase mutant and application thereof in preparing theophylline detection reagent |
CN202310217235.5A Pending CN116359146A (en) | 2019-01-09 | 2019-12-26 | Method for preparing conjugate |
CN201911372535.0A Active CN112285038B (en) | 2019-01-09 | 2019-12-27 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digitoxin detection reagent |
CN201911372147.2A Active CN112285037B (en) | 2019-01-09 | 2019-12-27 | 6-phosphoglucose dehydrogenase mutant and application thereof in preparing detection reagent |
CN202310811210.8A Pending CN116626281A (en) | 2019-01-09 | 2019-12-27 | Digitoxin detection kit |
CN202310811498.9A Pending CN116698772A (en) | 2019-01-09 | 2019-12-27 | Method for preparing conjugate |
CN202310811212.7A Pending CN116735512A (en) | 2019-01-09 | 2019-12-27 | Use of conjugates in the preparation of detection reagents |
CN202211151405.6A Pending CN115791649A (en) | 2019-01-09 | 2019-12-27 | Glycocholic acid detection kit |
CN202211151264.8A Pending CN116008201A (en) | 2019-01-09 | 2019-12-27 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of detection reagent |
CN202211153004.4A Pending CN115808398A (en) | 2019-01-09 | 2019-12-27 | Method for preparing conjugate |
CN202310726493.6A Pending CN116773795A (en) | 2019-01-09 | 2019-12-31 | Preparation method of conjugate |
CN202310726498.9A Pending CN116559472A (en) | 2019-01-09 | 2019-12-31 | Cortisol detection kit |
CN202310364266.3A Pending CN116718764A (en) | 2019-01-09 | 2019-12-31 | Use of conjugate in preparation of phenytoin detection reagent |
CN202310724066.4A Pending CN116773827A (en) | 2019-01-09 | 2019-12-31 | Use of conjugates in the preparation of detection reagents |
CN202310365160.5A Pending CN116355873A (en) | 2019-01-09 | 2019-12-31 | Preparation method of conjugate |
CN202310364997.8A Pending CN116298257A (en) | 2019-01-09 | 2019-12-31 | Phenytoin detection kit |
CN201911404154.6A Active CN111504920B (en) | 2019-01-09 | 2019-12-31 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of cortisol detection reagent |
CN201911403882.5A Active CN111650135B (en) | 2019-01-09 | 2019-12-31 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of phenytoin detection reagent |
CN202010000321.7A Active CN111487206B (en) | 2019-01-09 | 2020-01-02 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of vancomycin detection reagent |
CN202311025752.9A Pending CN117074335A (en) | 2019-01-09 | 2020-01-02 | Use of conjugates in the preparation of detection reagents |
CN202311025762.2A Pending CN117054643A (en) | 2019-01-09 | 2020-01-02 | Vancomycin detection kit |
CN202311025756.7A Pending CN117030640A (en) | 2019-01-09 | 2020-01-02 | Method for preparing conjugate |
CN202310811497.4A Pending CN116840468A (en) | 2019-01-09 | 2020-01-03 | Use of conjugates in the preparation of detection reagents |
CN202310810479.4A Pending CN116819060A (en) | 2019-01-09 | 2020-01-03 | Digoxin detection kit |
Family Applications After (25)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010004879.2A Active CN111487207B (en) | 2019-01-09 | 2020-01-03 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
CN202010009771.2A Active CN111504921B (en) | 2019-01-09 | 2020-01-06 | 6-glucose phosphate dehydrogenase mutant and application thereof in preparation of gentamicin detection reagent |
CN202310320137.4A Pending CN116148198A (en) | 2019-01-09 | 2020-01-06 | Preparation method of gentamicin detection reagent |
CN202310452740.8A Pending CN116430056A (en) | 2019-01-09 | 2020-01-06 | Method for preparing conjugate |
CN202310320729.6A Pending CN116297271A (en) | 2019-01-09 | 2020-01-06 | Use of conjugates in the preparation of kits |
CN202310452946.0A Pending CN116559425A (en) | 2019-01-09 | 2020-01-06 | Use of conjugates in the preparation of detection reagents |
CN202010009570.2A Active CN111537451B (en) | 2019-01-09 | 2020-01-06 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of tacrolimus detection reagent |
CN202310453290.4A Pending CN116338215A (en) | 2019-01-09 | 2020-01-06 | Tacrolimus detection kit |
CN202310318754.0A Pending CN116124721A (en) | 2019-01-09 | 2020-01-06 | Gentamicin detection kit |
CN202310508418.2A Pending CN116381253A (en) | 2019-01-09 | 2020-01-07 | Method for preparing conjugate |
CN202010013644.XA Active CN111487208B (en) | 2019-01-09 | 2020-01-07 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of methotrexate detection reagent |
CN202310554774.8A Pending CN116679047A (en) | 2019-01-09 | 2020-01-07 | Method for preparing conjugate |
CN202010013174.7A Active CN111678874B (en) | 2019-01-09 | 2020-01-07 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of cyclosporine A detection reagent |
CN202310507880.0A Pending CN116754756A (en) | 2019-01-09 | 2020-01-07 | Methotrexate detection kit |
CN202310508217.2A Pending CN116298330A (en) | 2019-01-09 | 2020-01-07 | Use of conjugates in the preparation of detection reagents |
CN202310555230.3A Pending CN116718761A (en) | 2019-01-09 | 2020-01-07 | Cyclosporine A detection kit |
CN202310553479.0A Pending CN116699125A (en) | 2019-01-09 | 2020-01-07 | Use of conjugates in the preparation of detection reagents |
CN202310702870.2A Pending CN116840462A (en) | 2019-01-09 | 2020-01-08 | Method for preparing conjugate |
CN202010017376.9A Active CN111537452B (en) | 2019-01-09 | 2020-01-08 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of amikacin detection reagent |
CN202310725902.0A Pending CN116577494A (en) | 2019-01-09 | 2020-01-08 | Use of conjugates in the preparation of detection reagents |
CN202010016535.3A Active CN111693473B (en) | 2019-01-09 | 2020-01-08 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of rapamycin detection reagent |
CN202310726230.5A Pending CN116577495A (en) | 2019-01-09 | 2020-01-08 | Method for preparing conjugate |
CN202310726069.1A Pending CN116754761A (en) | 2019-01-09 | 2020-01-08 | Amikacin detection kit |
CN202310702858.1A Pending CN116859035A (en) | 2019-01-09 | 2020-01-08 | Use of conjugates in the preparation of detection reagents |
CN202310702860.9A Pending CN116699122A (en) | 2019-01-09 | 2020-01-08 | Rapamycin detection kit |
Country Status (1)
Country | Link |
---|---|
CN (52) | CN110174363A (en) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110174363A (en) * | 2019-01-09 | 2019-08-27 | 北京九强生物技术股份有限公司 | Glucose-6-phosphate dehydrogenase mutant and its purposes in preparation detection reagent |
CN113046335B (en) * | 2019-12-27 | 2023-05-26 | 中国科学院天津工业生物技术研究所 | Bionic coenzyme-favored glucose 6-phosphate dehydrogenase mutant and application thereof |
JP2023509037A (en) * | 2020-01-07 | 2023-03-06 | 北京九▲強▼生物技▲術▼股▲フン▼有限公司 | Glucose-6-phosphate dehydrogenase variants and their use in preparing detection reagents |
CN112114127A (en) * | 2020-09-09 | 2020-12-22 | 武汉生之源生物科技股份有限公司 | Glycocholic acid homogeneous enzyme immunoassay kit and preparation method and application thereof |
CN112225795A (en) * | 2020-10-14 | 2021-01-15 | 湖南苏阳医疗科技有限公司 | 6-hydroxy sulfate melatonin derivative and preparation method and application of immunogen and specific antibody thereof |
CN112574969A (en) * | 2020-12-28 | 2021-03-30 | 郑州伊美诺生物技术有限公司 | G6PDH mutant and application thereof |
CN113567662A (en) * | 2021-07-08 | 2021-10-29 | 重庆中元汇吉生物技术有限公司 | Kit for determining glycocholic acid and preparation method thereof |
CN113736744B (en) * | 2021-10-14 | 2023-07-18 | 江南大学 | Digitoxin monoclonal antibody hybridoma cell strain and application thereof |
CN115236216B (en) * | 2022-06-07 | 2024-03-01 | 合肥和合医疗科技有限公司 | Kit for detecting immunosuppressant in whole blood by high performance liquid chromatography tandem mass spectrometry, preparation method and detection method thereof |
Family Cites Families (62)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2116301B1 (en) * | 1970-12-07 | 1974-08-30 | Brun Lab Sa Le | |
US4190496A (en) * | 1971-05-14 | 1980-02-26 | Syva Company | Homogeneous enzyme assay for antibodies |
US3997525A (en) * | 1974-01-16 | 1976-12-14 | Bio-Tec, Inc. | Tetra-125 iodo-di-tyramine of digitalis derivative and process for making the same |
DE2901218A1 (en) * | 1979-01-13 | 1980-07-17 | Byk Gulden Lomberg Chem Fab | THEOPHYLLIN |
JPS5618983A (en) * | 1979-07-25 | 1981-02-23 | Eisai Co Ltd | Theophylline derivative and its preapration |
US4262089A (en) * | 1980-04-07 | 1981-04-14 | Syva Company | Theophylline antigens and antibodies |
US4341866A (en) * | 1980-06-02 | 1982-07-27 | Syva Company | Antienzyme termination in enzyme immunoassays |
JPS57178159A (en) * | 1981-04-27 | 1982-11-02 | Banyu Pharmaceut Co Ltd | Chemical reagent for detection of amicacin and its determination |
US4608336A (en) * | 1981-08-27 | 1986-08-26 | Miles Laboratories, Inc. | #3B theophylline immunoassay employing 9-theophylline reagents |
US4469797A (en) * | 1982-09-23 | 1984-09-04 | Miles Laboratories, Inc. | Digoxigenin immunogens, antibodies, labeled conjugates, and related derivatives |
EP0119767B1 (en) * | 1983-03-11 | 1990-11-22 | FUJIREBIO KABUSHIKI KAISHA also trading as FUJIREBIO INC. | Method of measuring ligands |
IT1199088B (en) * | 1984-03-09 | 1988-12-30 | Miles Italiana | SPECIFIC BOND TEST BY USING ANTI-G6PDH AS A MARKER |
US4622294A (en) * | 1985-02-08 | 1986-11-11 | Kung Viola T | Liposome immunoassay reagent and method |
US5068198A (en) * | 1986-03-26 | 1991-11-26 | Syntex (U.S.A.) Inc. | Liquid single reagent for assays involving confining gels |
EP0399127A1 (en) * | 1989-05-23 | 1990-11-28 | Pharmacia ENI Diagnostics Inc. | Homogeneous immunochemical method for determining haptens by means of ion selective electrodes |
DE3919915A1 (en) * | 1989-06-19 | 1990-12-20 | Boehringer Mannheim Gmbh | AMINOALKYLMALEIMIDES AND DERIVED HAPTEN AND ANTIGEN DERIVATIVES AND CONJUGATES WITH PEPTIDES OR PROTEINS |
JPH0833394B2 (en) * | 1990-10-03 | 1996-03-29 | 三洋化成工業株式会社 | Method for producing enzyme-labeled hapten |
CA2055812A1 (en) * | 1990-11-20 | 1992-05-21 | Carl N. Skold | Method of stabilizing enzyme conjugates |
ES2181734T3 (en) * | 1990-11-20 | 2003-03-01 | Dade Behring Marburg Gmbh | IMMUNOLOGICAL TEST OF CYCLOSPORINE. |
CA2087397A1 (en) * | 1992-01-22 | 1993-07-23 | Kazuhisa Kubotsu | Immunoassay and reagents used therefor |
CA2156397C (en) * | 1993-04-08 | 2007-05-15 | Valerie Quesniaux | Rapamycin assay |
US6455288B1 (en) * | 1993-04-08 | 2002-09-24 | Dade Behring Marburg Gmbh | Homogeneous immunoassays using mutant glucose-6-phosphate dehydrogenases |
US5747352A (en) * | 1994-05-23 | 1998-05-05 | Beckman Instruments, Inc. | Reagents and methods for the rapid and quantitative assay of pharmacological agents |
WO2000022000A1 (en) * | 1998-10-09 | 2000-04-20 | Isotechnika, Inc. | Methods for the production of antibodies to specific regions of cyclosporine and cyclosporine metabolites |
AUPP751398A0 (en) * | 1998-12-04 | 1999-01-07 | Commonwealth Scientific And Industrial Research Organisation | Methotrexate derivatives |
US7078495B1 (en) * | 1999-08-03 | 2006-07-18 | Dade Behring Inc. | Monoclonal antibodies to tacrolimus and immunoassay methods for tacrolimus |
US6653456B2 (en) * | 2001-07-31 | 2003-11-25 | Roche Diagnostics Corporation | Site-specific aminoglycoside derivatives and their use in immunodiagnostic assays |
US20050176080A1 (en) * | 2004-02-10 | 2005-08-11 | Vani Bodepudi | Hapten, immunogens and derivatives of ascomycin useful for preparation of antibodies and immunoassays |
US20060046273A1 (en) * | 2004-08-27 | 2006-03-02 | Lin-Zhi International Inc. | Homogeneous enzyme immunoassay for oral fluid |
TW200635954A (en) * | 2004-12-17 | 2006-10-16 | Isotechnika Inc | Metabolites of cyclosporin analogs |
US7189582B2 (en) * | 2005-04-27 | 2007-03-13 | Dade Behring Inc. | Compositions and methods for detection of sirolimus |
JP4746926B2 (en) * | 2005-06-29 | 2011-08-10 | シスメックス株式会社 | Glucose-6-phosphate dehydrogenase-containing reagent and glucose-6-phosphate dehydrogenase stabilization method |
CN101638640B (en) * | 2009-09-07 | 2011-01-12 | 北京利德曼生化股份有限公司 | Glucose-6-phosphoric acid dehydrogenase and nucleotide sequence, recombinant vector, recombinant host cell and kit thereof |
JP2014503197A (en) * | 2010-11-24 | 2014-02-13 | ディーエイチ テクノロジーズ デベロップメント プライベート リミテッド | High-throughput, sensitive detection of glucose-6-phosphate dehydrogenase |
CN102565399B (en) * | 2010-12-07 | 2015-06-03 | 北京望尔生物技术有限公司 | Method for detecting hydrocortisone and special enzyme-linked immunosorbent assay kit thereof |
CN102807618A (en) * | 2011-08-10 | 2012-12-05 | 重庆金域医学检验所有限公司 | Preparation method of phenytoin homogeneous enzyme immunoassay kit and phenytoin polyclonal antibodies |
JP5896375B2 (en) * | 2011-09-09 | 2016-03-30 | 池田食研株式会社 | Modified glucose dehydrogenase gene |
CN102424829B (en) * | 2011-10-26 | 2013-10-16 | 苏州汉酶生物技术有限公司 | Method for synthesizing temsirolimus through enzyme catalysis |
US8771964B2 (en) * | 2012-02-02 | 2014-07-08 | Siemens Healthcare Diagnostics Inc. | Compositions and methods for detection of methadone metabolite |
CN104619350A (en) * | 2012-06-14 | 2015-05-13 | Ambrx公司 | Anti-psma antibodies conjugated to nuclear receptor ligand polypeptides |
CN103242446A (en) * | 2012-07-25 | 2013-08-14 | 苏州博源医疗科技有限公司 | Theophylline immunogen and preparation method and application thereof |
CN102757391B (en) * | 2012-08-01 | 2015-08-26 | 苏州博源医疗科技有限公司 | A kind of Phenobarbital derivatives and its preparation method and application |
EP3083657B1 (en) * | 2013-12-17 | 2022-01-26 | Siemens Healthcare Diagnostics Inc. | Preparation of multi-hapten mutant g6pdh conjugates and their use for detection of multiple analytes |
CN104016923B (en) * | 2014-01-08 | 2016-08-31 | 南开大学 | Phenytoin derivant and its production and use |
CN103739703B (en) * | 2014-02-11 | 2015-07-15 | 苏州博源医疗科技有限公司 | Glycocholic acid immunogen, anti-glycocholic acid specific antibody and detection reagent |
CN103760348B (en) * | 2014-02-11 | 2015-03-11 | 苏州博源医疗科技有限公司 | Glycocholic acid immunodetection reagent and preparing method and detecting method thereof |
JP6398295B2 (en) * | 2014-04-30 | 2018-10-03 | ニプロ株式会社 | Mutant glucose-6-phosphate dehydrogenase |
CN104447745B (en) * | 2014-11-06 | 2016-03-30 | 济南金域医学检验中心有限公司 | A kind of theophylline homogeneous enzyme immunoassay detects tests test kit and preparation method thereof |
CN104569373B (en) * | 2015-01-27 | 2016-08-17 | 苏州博源医疗科技有限公司 | A kind of methotrexate homogeneous enzyme immunoassay detectable and preparation thereof and detection method |
US10330683B2 (en) * | 2015-02-04 | 2019-06-25 | Genentech, Inc. | Mutant smoothened and methods of using the same |
CN105131105A (en) * | 2015-07-27 | 2015-12-09 | 苏州博源医疗科技有限公司 | Cortisol immunogen, derivative, antibody, detection reagent and preparation method |
CN106565809B (en) * | 2016-07-08 | 2018-05-01 | 北京九强生物技术股份有限公司 | A kind of enzyme donor conjugate of beta galactosidase and its purposes in glycocholic acid detection |
CN106226512B (en) * | 2016-07-29 | 2018-10-16 | 胡清 | A kind of detection method of kit, the preparation method of kit and the peripheral blood glycocholic acid realized using the kit |
CN106190996B (en) * | 2016-08-30 | 2019-05-21 | 美康生物科技股份有限公司 | A kind of G 6 PD mutant |
CN109797143B (en) * | 2016-09-22 | 2021-02-12 | 北京九强生物技术股份有限公司 | Reagent containing escherichia coli beta galactosidase receptor |
CN106872681B (en) * | 2017-01-23 | 2019-11-19 | 四川精卫食品检测科技有限公司 | Amikacin and the two-in-one quick detection enzyme linked immunological kit of kanamycins and its application |
CN108593905A (en) * | 2017-12-22 | 2018-09-28 | 太原瑞盛生物科技有限公司 | A kind of digoxin immune detection reagent and its preparation and detection method |
CN108586562B (en) * | 2018-05-08 | 2019-09-06 | 苏州博源医疗科技有限公司 | A kind of cortex 01 derivatives and the preparation method and application thereof |
CN108717117A (en) * | 2018-05-23 | 2018-10-30 | 太原瑞盛生物科技有限公司 | A kind of vancomycin immunologic function test reagent and its preparation and detection method |
CN109111494A (en) * | 2018-08-30 | 2019-01-01 | 苏州博源医疗科技有限公司 | Derivatives of estradiol, immunogene, antibody, enzyme mark conjugate, detection reagent and preparation method thereof |
CN110174363A (en) * | 2019-01-09 | 2019-08-27 | 北京九强生物技术股份有限公司 | Glucose-6-phosphate dehydrogenase mutant and its purposes in preparation detection reagent |
CN112574969A (en) * | 2020-12-28 | 2021-03-30 | 郑州伊美诺生物技术有限公司 | G6PDH mutant and application thereof |
-
2019
- 2019-05-21 CN CN201910423122.4A patent/CN110174363A/en active Pending
- 2019-12-26 CN CN202310257027.8A patent/CN116144619A/en active Pending
- 2019-12-26 CN CN201911365439.3A patent/CN111239060B/en active Active
- 2019-12-26 CN CN202310217235.5A patent/CN116359146A/en active Pending
- 2019-12-27 CN CN201911372535.0A patent/CN112285038B/en active Active
- 2019-12-27 CN CN201911372147.2A patent/CN112285037B/en active Active
- 2019-12-27 CN CN202310811210.8A patent/CN116626281A/en active Pending
- 2019-12-27 CN CN202310811498.9A patent/CN116698772A/en active Pending
- 2019-12-27 CN CN202310811212.7A patent/CN116735512A/en active Pending
- 2019-12-27 CN CN202211151405.6A patent/CN115791649A/en active Pending
- 2019-12-27 CN CN202211151264.8A patent/CN116008201A/en active Pending
- 2019-12-27 CN CN202211153004.4A patent/CN115808398A/en active Pending
- 2019-12-31 CN CN202310726493.6A patent/CN116773795A/en active Pending
- 2019-12-31 CN CN202310726498.9A patent/CN116559472A/en active Pending
- 2019-12-31 CN CN202310364266.3A patent/CN116718764A/en active Pending
- 2019-12-31 CN CN202310724066.4A patent/CN116773827A/en active Pending
- 2019-12-31 CN CN202310365160.5A patent/CN116355873A/en active Pending
- 2019-12-31 CN CN202310364997.8A patent/CN116298257A/en active Pending
- 2019-12-31 CN CN201911404154.6A patent/CN111504920B/en active Active
- 2019-12-31 CN CN201911403882.5A patent/CN111650135B/en active Active
-
2020
- 2020-01-02 CN CN202010000321.7A patent/CN111487206B/en active Active
- 2020-01-02 CN CN202311025752.9A patent/CN117074335A/en active Pending
- 2020-01-02 CN CN202311025762.2A patent/CN117054643A/en active Pending
- 2020-01-02 CN CN202311025756.7A patent/CN117030640A/en active Pending
- 2020-01-03 CN CN202310811497.4A patent/CN116840468A/en active Pending
- 2020-01-03 CN CN202310810479.4A patent/CN116819060A/en active Pending
- 2020-01-03 CN CN202310810455.9A patent/CN116840467A/en active Pending
- 2020-01-03 CN CN202010004879.2A patent/CN111487207B/en active Active
- 2020-01-06 CN CN202010009771.2A patent/CN111504921B/en active Active
- 2020-01-06 CN CN202310320137.4A patent/CN116148198A/en active Pending
- 2020-01-06 CN CN202310452740.8A patent/CN116430056A/en active Pending
- 2020-01-06 CN CN202310320729.6A patent/CN116297271A/en active Pending
- 2020-01-06 CN CN202310452946.0A patent/CN116559425A/en active Pending
- 2020-01-06 CN CN202010009570.2A patent/CN111537451B/en active Active
- 2020-01-06 CN CN202310453290.4A patent/CN116338215A/en active Pending
- 2020-01-06 CN CN202310318754.0A patent/CN116124721A/en active Pending
- 2020-01-07 CN CN202310508418.2A patent/CN116381253A/en active Pending
- 2020-01-07 CN CN202010013644.XA patent/CN111487208B/en active Active
- 2020-01-07 CN CN202310554774.8A patent/CN116679047A/en active Pending
- 2020-01-07 CN CN202010013174.7A patent/CN111678874B/en active Active
- 2020-01-07 CN CN202310507880.0A patent/CN116754756A/en active Pending
- 2020-01-07 CN CN202310508217.2A patent/CN116298330A/en active Pending
- 2020-01-07 CN CN202310555230.3A patent/CN116718761A/en active Pending
- 2020-01-07 CN CN202310553479.0A patent/CN116699125A/en active Pending
- 2020-01-08 CN CN202310702870.2A patent/CN116840462A/en active Pending
- 2020-01-08 CN CN202010017376.9A patent/CN111537452B/en active Active
- 2020-01-08 CN CN202310725902.0A patent/CN116577494A/en active Pending
- 2020-01-08 CN CN202010016535.3A patent/CN111693473B/en active Active
- 2020-01-08 CN CN202310726230.5A patent/CN116577495A/en active Pending
- 2020-01-08 CN CN202310726069.1A patent/CN116754761A/en active Pending
- 2020-01-08 CN CN202310702858.1A patent/CN116859035A/en active Pending
- 2020-01-08 CN CN202310702860.9A patent/CN116699122A/en active Pending
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111487207B (en) | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
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 |