Structure of 133081-26-2
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CAS No. : | 133081-26-2 |
Formula : | C15H18N4O6 |
M.W : | 350.33 |
SMILES Code : | CC(C)(C)OC(=O)NNC1=NC=C(C=C1)C(=O)ON1C(=O)CCC1=O |
MDL No. : | MFCD03788694 |
InChI Key : | PHWZJVJAIAVOAP-UHFFFAOYSA-N |
Pubchem ID : | 15343047 |
GHS Pictogram: |
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Signal Word: | Warning |
Hazard Statements: | H315-H319-H335 |
Precautionary Statements: | P261-P305+P351+P338 |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 2h; | Dimethylformamide (DMF) (2 ml) and 60 mul of diisopropylethylamine is added to 186 mg of N-For-Met-Leu-Phe-diaminohexyl amide followed by 154 mg succinimidyl-6-t-BOC-hydrazinopyridine-3-carboxylic acid in 1 ml DMF. The mixture becomes yellow and the peptide dissolves within a short time. After 2 hours, ether is added to the reaction mixture and the upper layer is discarded. Water is added to the oily residue causing a solid to form. The solid is washed with 5% sodium bicarbonate, water and ethyl acetate, and the yield is determined. The t-BOC protecting group is removed by stirring the crude product with 5 ml of trifluoroacetic acid (TFA) containing 0.1 ml of p-cresol for 15 min. at 20 C. Prolonged treatment with TFA results in increased levels of a side product. The TFA is removed by rotary evaporation, and ether is added to the residue to precipitate the deprotected peptide. The product is purified by reverse phase HPLC on a 2.5×50 cm Whatman ODS-3 column is eluted with a gradient of acetonitrile in 0.1% TFA. Fractions containing the major component is combined and the solvent is removed to yield the desired product. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With phosphate buffer; In tetrahydrofuran; at 20℃; for 4h;pH 7.5; | In a 25-mL round bottomed flask, 50 mg of carboxyl-polyethylene glycol-amine (0.0147 mmole) (Shearwater) is diluted with 2.5 ml of 10 mM phosphate buffer, pH 7.5. To the resulting solution is added 15.5 mg of N-hydroxysuccinimidyl-6-BOC-hydrazinonicotinate (Solulink) in tetrahydrofuran (THF). The resulting reaction is stirred at room temperature under argon in the dark. After 4 hours, the aqueous phase is extracted with dichloromethane (DCM). The DCM phase is dried over MgSO4 and concentrated in vacuo to provide a liquid residue which is then precipitated using ether. The precipitated PEG-polymer is collected by filtration, and washed with 10 ml of cold (-20 C.) isopropyl alcohol (IPA) to provide carboxy-PEG-6-BOC-hydrazinonicotinate (CPN-Boc) which is then diluted with 1 ml of THF. To the resulting solution is added 15 mg of N,N'-dicyclohexylcarbodiimide (DCC), and 10.2 mg of N-hydroxysuccinimide (NHS) and the reaction mixture is stirred under argon for 3 hours. Dry silica gel (1 mg) is added to the solution and allowed to settle. The supernatant is separated from the solid precipitate and the solvent is then removed under vacuum to provide a residue which is resuspended in 0.5 ml of ethyl acetate, extracted with 0.5 ml of 3M HCl for 10 minutes (3 times). The organic phase is dried over MgSO4, concentrated in vacuo and extracted with ether. Cold IPA is then added to the ethereal solution to provide compound 2 as a precipitate. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
In 1,4-dioxane; water; sodium hydrogencarbonate; | 6-(BOC-Hydrazine)nicotinamido-S-(2-thio-5-nitropyridyl)-(L)-cysteine S-(2-Thio-5-nitropyridyl)-(L)-cysteine hydrochloride (220 mg, 0.70 mmol) was dissolved in saturated aqueous sodium bicarbonate (5 ml) and a solution of succinimidyl 6-(BOC-hydrazino)nicotinate (246 g, 0.70 mmol) in dioxane (5 ml) was added and the reaction mixture was stirred at room temperature for 3 hours. Water (25 ml) was added and the aqueous solution was washed with ethyl acetate to remove unreacted ester. The aqueous phase was acidified to pH 3.3 with 1N HCl then saturated with sodium chloride. The acidic aqueous solution was extracted with ethyl acetate (2*35 ml). The combined organic extracts were dried over magnesium sulphate, filtered and concentrated to give 145 mg of a yellow solid which was suspended in ether and isolated by filtration to give 35 mg of the desired product. The filtrate was concentrated and treated with ether/hexanes to yield a further 95 mg of desired product; total yield 125 mg (35%). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Subsequent to the automated assembly the peptide resin (13 mg, 0.01 mmole) was transferred to a Microwave vial and Boc-HYNIC-NHS ester (5mg, 0.015 mmoleJ/HOBt (2 mg, 0.015 mmole}/DIEA (8 mul, 0.0.6 mmole) in DMF (3 ml) was added. The resin mix was left standing for 15 min at RT and thereafter placed in a microwave for 2 h at 6O0C. The 20 resin was cleaved in TFA/TIS/H2O (10:0.25:0.25) for 2h then filtrated, triturated in diethylether, rotavapory evaporated and lyophilised. The crude peptide was analysed by LC-MS (20-40%B in 20 min). The analysis demonstrated the presence of two main peaks. The crude peptide mix was purified by prep HPLC (20-35% B in 40 min). One- - fraction "was collected, analysed and lyophfliseaTVield: 1 mg of white solid. HPLC purity: 25 98%. ESI-MS: Theoretical MW: 1857.2. Obtained (M+H)2+/2: 929.5. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
74% | With triethylamine; In dichloromethane; at 20℃; for 1h; | EXAMPLE 15[0115] Synthesis of 2-{3-[l-carboxy-5-({6-[N'-(4-fluoro-benzylidene)-hydrazino]- pyridine- 3-carbonyl}-amino)-pentyl]-ureido}-pentanedioic acid (15).; To a solution of compound 1 (0.040 g, 0.047 mmol) in CH2Cl2 (2 mL) was added triethylamine (0.020 mL, 0.14 mmol), followed by succinimidyl 6-(N'-tert-butoxycarbonyl-hydrazino)-nicotinate (0.020 g, 0.057 mmol, Bioconjugate Chem. 1991, 2, 333-336). After stirred for 1 hour at room temperature, the solvent was evaporated. The crude material was purified by a silica column using methanol/methylene chloride (10:90) to afford 0.032 g (74%) of compound 16. 1H NMR (400 MHz, CDCl3) £8.54 (s, IH), 7.90 (m, IH), 7.17-7.23 (m, 6H), 6.90-7.05 (m, 3H), 6.79- 6.84 (m, 6H), 6.55 (d, J= 8.8 Hz, 2H), 5.79 (m, 2H), 4.94-5.07 (m, 6H), 4.38-4.45 (m, 2H), 3.74 (m, 9H), 3.26 (m, 2H), 2.33 (m, 2H), 2.07 (m, IH), 1.85 (m, IH), 1.68 (m, IH), 1.18-1.55 (m, 13H). ESI-Mass calcd for C47H59N6O13 [M + H]+ 915.4 , found 914.9. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With sodium borate; In acetonitrile; at 4℃;pH 8.5; | TheLysine conjugate 7 (51.0 mg, 0.06mmol) was then treated with Boc-HYNIC-OSu (21.0 mg, 0.06 mmol) (Solulink, San Diego, CA).) in 2 mL acetonitrile-sodium borate buffer (0.1N, pH 8.5) (50/50, v/v) and incubated at 4 C overnight,followed by concentration under reduced pressure and TFA deprotection (1 mL TFAin 2 mL anhydrous CH2Cl2 |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
84% | With dicyclohexyl-carbodiimide; In N,N-dimethyl-formamide; at 25℃; for 17h;Inert atmosphere; | To a solution of 6-Boc-hydrazinopyridine-3-carboxylicacid 4b (0.57 g, 2.26 mmol) and N-hydrosuccinimide (0.27g, 2.26 mmol) in DMF DCC (0.47 g, 2.26 mmol) was added.The reaction mixture became cloudy after 1h. After 16h, thereaction mixture was concentrated to dryness and the residuewas purified by chromatography on silica gel with ethyl acetateas eluant to give a white solid (0.66 g, 84%). Mp =174C. IR (ATR, cm-1) 3321; 2979; 1729; 1594; 1365; 1240;1198; 1068; 974; 641. 1H NMR (DMSO) 9.46 (s, 1H), 9.20(s, 1H), 8.76 (d, 1H, J= 2.1 Hz), 8.11 (d, 1H, J= 8.1 Hz),6.67 (d, 1H, J= 9 Hz), 2.88 (s, 4H), 1.44 (s, 9H). 13C NMR(DMSO) 172.8; 170.5; 160.9; 155.4; 151.8; 110.0; 79.6;28.0; 25.5. MS (ESI+) [M+H]: 351.00. HRMS (ESI+)[M+H]+: 351.1299 observed, 351.1305 calculated forC15H19N4O6. |
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