Tromethamine hydrochloride → Citrate
Reaction equation
C(O)[C:2](N)([CH2:5][OH:6])[CH2:3][OH:4].Cl.C(SCCNC(=O)CCNC(=O)[C@H](O)C(C)(C)COP(O)(=O)OP(O)(=O)OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1OP(O)(O)=O)(=[O:18])CCCCCCC.[CH3:67][CH2:68][CH2:69][CH2:70][CH2:71][CH2:72][CH2:73][C:74]([O:76][CH2:77][C@H:78]([NH:285][C:286]([CH2:288][NH:289][C:290]([C@@H:292]([NH2:300])[CH2:293][CH2:294][CH2:295][NH:296][C:297]([NH2:299])=[NH:298])=[O:291])=[O:287])[C:79]([NH:81][C@H:82]([C:85]([NH:87][C@H:88]([C:96]([NH:98][C@H:99]([C:104]([NH:106][C@H:107]([C:110]([N:112]1[C@H:116]([C:117]([NH:119][C@H:120]([C:126]([NH:128][C@H:129]([C:136]([NH:138][C@H:139]([C:145]([NH:147][C@H:148]([C:154]([NH:156][C@H:157]([C:159]([NH:161][C@H:162]([C:168]([NH:170][C@H:171]([C:177]([NH:179][C@H:180]([C:188]([NH:190][C@H:191]([C:197]([NH:199][C@H:200]([C:206]([NH:208][C@H:209]([C:212]([NH:214][C@H:215]([C:221]([NH:223][C@H:224]([C:230]([N:232]2[C@H:236]([C:237]([N:239]3[C@H:243]([C:244]([NH:246][C@H:247]([C:249]([NH:251][C@H:252]([C:258]([NH:260][C@H:261]([C:266]([NH:268][C@H:269]([C:275]([N:277]4[C@H:281]([C:282]([OH:284])=[O:283])[CH2:280][CH2:279][CH2:278]4)=[O:276])[CH2:270][CH2:271][C:272]([NH2:274])=[O:273])=[O:267])[CH2:262][CH:263]([CH3:265])[CH3:264])=[O:259])[CH2:253][CH2:254][CH2:255][CH2:256][NH2:257])=[O:250])[CH3:248])=[O:245])[CH2:242][CH2:241][CH2:240]3)=[O:238])[CH2:235][CH2:234][CH2:233]2)=[O:231])[CH2:225][CH2:226][CH2:227][CH2:228][NH2:229])=[O:222])[CH2:216][CH2:217][CH2:218][CH2:219][NH2:220])=[O:213])[CH2:210][OH:211])=[O:207])[CH2:201][CH2:202][C:203]([OH:205])=[O:204])=[O:198])[CH2:192][CH2:193][CH2:194][CH2:195][NH2:196])=[O:189])[CH2:181][CH2:182][CH2:183][NH:184][C:185]([NH2:187])=[NH:186])=[O:178])[CH2:172][CH2:173][C:174]([NH2:176])=[O:175])=[O:169])[CH2:163][CH2:164][C:165]([NH2:167])=[O:166])=[O:160])[CH3:158])=[O:155])[CH2:149][CH2:150][CH2:151][CH2:152][NH2:153])=[O:146])[CH2:140][CH2:141][C:142]([NH2:144])=[O:143])=[O:137])[CH2:130][C:131]2[NH:135][CH:134]=[N:133][CH:132]=2)=[O:127])[CH2:121][CH2:122][C:123]([OH:125])=[O:124])=[O:118])[CH2:115][CH2:114][CH2:113]1)=[O:111])[CH2:108][OH:109])=[O:105])[CH2:100][CH:101]([CH3:103])[CH3:102])=[O:97])[CH2:89][C:90]1[CH:91]=[CH:92][CH:93]=[CH:94][CH:95]=1)=[O:86])[CH2:83][OH:84])=[O:80])=[O:75].[OH:301][C:302](CCCC[C@H]1[C@@H]2[C@@H](NC(N2)=O)CS1)=[O:303]>CS(C)=O>[C:74]([O-:76])(=[O:75])[CH2:73][C:5]([CH2:2][C:3]([O-:4])=[O:18])([C:302]([O-:303])=[O:301])[OH:6].[CH3:67][CH2:68][CH2:69][CH2:70][CH2:71][CH2:72][CH2:73][C:74]([O:76][CH2:77][C@H:78]([NH:285][C:286]([CH2:288][NH:289][C:290]([C@@H:292]([NH2:300])[CH2:293][CH2:294][CH2:295][NH:296][C:297]([NH2:299])=[NH:298])=[O:291])=[O:287])[C:79]([NH:81][C@H:82]([C:85]([NH:87][C@H:88]([C:96]([NH:98][C@H:99]([C:104]([NH:106][C@H:107]([C:110]([N:112]1[C@H:116]([C:117]([NH:119][C@H:120]([C:126]([NH:128][C@H:129]([C:136]([NH:138][C@H:139]([C:145]([NH:147][C@H:148]([C:154]([NH:156][C@H:157]([C:159]([NH:161][C@H:162]([C:168]([NH:170][C@H:171]([C:177]([NH:179][C@H:180]([C:188]([NH:190][C@H:191]([C:197]([NH:199][C@H:200]([C:206]([NH:208][C@H:209]([C:212]([NH:214][C@H:215]([C:221]([NH:223][C@H:224]([C:230]([N:232]2[C@H:236]([C:237]([N:239]3[C@H:243]([C:244]([NH:246][C@H:247]([C:249]([NH:251][C@H:252]([C:258]([NH:260][C@H:261]([C:266]([NH:268][C@H:269]([C:275]([N:277]4[C@H:281]([C:282]([OH:284])=[O:283])[CH2:280][CH2:279][CH2:278]4)=[O:276])[CH2:270][CH2:271][C:272]([NH2:274])=[O:273])=[O:267])[CH2:262][CH:263]([CH3:264])[CH3:265])=[O:259])[CH2:253][CH2:254][CH2:255][CH2:256][NH2:257])=[O:250])[CH3:248])=[O:245])[CH2:242][CH2:241][CH2:240]3)=[O:238])[CH2:235][CH2:234][CH2:233]2)=[O:231])[CH2:225][CH2:226][CH2:227][CH2:228][NH2:229])=[O:222])[CH2:216][CH2:217][CH2:218][CH2:219][NH2:220])=[O:213])[CH2:210][OH:211])=[O:207])[CH2:201][CH2:202][C:203]([OH:205])=[O:204])=[O:198])[CH2:192][CH2:193][CH2:194][CH2:195][NH2:196])=[O:189])[CH2:181][CH2:182][CH2:183][NH:184][C:185]([NH2:187])=[NH:186])=[O:178])[CH2:172][CH2:173][C:174]([NH2:176])=[O:175])=[O:169])[CH2:163][CH2:164][C:165]([NH2:167])=[O:166])=[O:160])[CH3:158])=[O:155])[CH2:149][CH2:150][CH2:151][CH2:152][NH2:153])=[O:146])[CH2:140][CH2:141][C:142]([NH2:144])=[O:143])=[O:137])[CH2:130][C:131]2[NH:135][CH:134]=[N:133][CH:132]=2)=[O:127])[CH2:121][CH2:122][C:123]([OH:125])=[O:124])=[O:118])[CH2:115][CH2:114][CH2:113]1)=[O:111])[CH2:108][OH:109])=[O:105])[CH2:100][CH:101]([CH3:103])[CH3:102])=[O:97])[CH2:89][C:90]1[CH:91]=[CH:92][CH:93]=[CH:94][CH:95]=1)=[O:86])[CH2:83][OH:84])=[O:80])=[O:75]Participants
Reactant 3
Product 2
Reagents, catalysts & solvents 1
Procedure
A compound solution (2 μL) diluted with an assay buffer containing 50 mM Tris-HCl (pH 7.5), 0.5% TWEEN20, and Complete to 15% DMSO and 90 μg/mL human GOAT-expressing microsome fraction (2 μL) were incubated at room temperature for 20 min, after which a substrate mixed solution (2 μL) containing 30 μM octanoyl-CoA (CHEM-IMPLEX) and 3 μM Ghrelin-biotin (Gly-Ser-Ser-Phe-Leu-Ser-Pro-Glu-His-Gln-Arg-Val-Gln-Gln-Arg-Lys-Glu-Ser-Lys-Lys-Pro-Pro-Ala-Lys-Leu-Gln-Pro-Arg-epsilon aminocaproic acid-biotin (Greiner Coutom synthesis)) was added to 384 white small volume plate (Greiner). After reaction for 20 min, 2 μL of citrate buffer (pH3) (mixed solution of 0.1 M citric acid (39.8 mL) and 0.2 M Na2HPO4 (10.2 mL)), 2 μL of 25 nM anti-active ghrelin antibody (produced in mouse), 5 μL of Eu(K)-Anti Mouse IgG Pab (Cis Bio) 100-fold diluted with HTRF detection buffer (Cis Bio) and 5 μL of streptavidin-Xlent (Cis Bio) 66.6-fold diluted with HTRF detection buffer (Cis Bio) were added. After incubation at room temperature for 1 hr or longer, the fluorescence values at 620 nm and 665 nm were measured by Envision (PerkinElmer). The data was calculated [(665 nm signal/620 nm signal)×104], and the inhibitory activity against human GOAT was shown by the data where 0% control was without compound, and 100% control was without enzyme. The results are shown in Table 34.
Workup
- additionwas added to 384 white small volume plate (Greiner)
- customAfter reaction for 20 min
Sources & evidence
- ORD record
- ord-865c6c65e89548928efc8807ca1aa113
- Source dataset
- uspto-grants-2016_01
- Patent number
- US09238639B2
Reaction notes
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