NHS-LC-Biotin

Biotin-C5-NHS Ester is an alkyl/ether-based PROTAC linker that can be used in the synthesis of PROTACs[1].

Synthesis of Biotin-T-Epitope Peptide 27:Next, a mixture of EZ-Link® NHS-Biotin reagent (13 (16.9 mg, 0.0359 mmol), EZ-Link' NHS-LC-Biotin (17 .9 mg, 0.0394mmol) and DIEA (9.3 mg, l2.5f.IL, 0.0718 mmol) in DMF (0.5 mL) was stirred at rt for l h. Thereaction mixture was concentrated under reduced pressure and the resulting residue was purified bypreparatory TLC (CH2Cl2:MeOH-NH3 (7N), 10: I) to give 20.8 mg (72percent) ofPU-H71-biotin4. 1HNMR (500 MHz, CDC, ) o 8.22 (s, H), 7.52 (t, J = 5.6 Hz, IH), 7.36 (s, H), 7.03 (s, H), 6.66 (t, J= 5.5 Hz, H), 6.25 (br s, 2H), 6.03 (s, 2H), 4.47-4.52 (m, H), 4.28-4.33 (m, H), 4.25 (t, J = 6.8 Hz, 2H), 3.17-3.25 (m, 4H), 3.11-3.17 (m, IH), 2.90 (dd, J= 5.0, 12.9 Hz, H), 2.63-2.79 (m, H), 2.24 (t, J= 7.4 Hz, 2H), 2.13-2.19 (m, 2H), 1.94-2.02 (m, 2H), 1.58-1.74 (m, 6H), 1.48-1.56 (m, 2H), 1.31-1.46 (m, 4H); MS (ESI): mlz 810.3 [M+Ht.2 (15 mg, 0.0292 mmol), EZ-Link' NHS-LC-Biotin (14.6 mg, 0.0321mmol) and DIEA (7.5 mg, 10.2f.IL, 0.0584 mmol) in DMF (0.5 mL) was heated at 35 °C for 6 h. Thereaction mixture was concentrated under reduced pressure and the resulting residue was purified bypreparatoryTLC (CH2Cl2:MeOH-NH, (7N), 10:1) to give 10.3 mg (41percent) ofPU-H71-biotin7.lnaddition, 6.9 mg ofunreacted 2 was recovered to give an actual yield of77percent. 1H NMR (500 MHz, CDCh, 2 rotamers) o 8.26-8.29 (m, H), 7.29 (s, 0.4H), 7.28 (s, 0.6H), 6.87 (s, 0.4H), 6.85 (s, 0.6H), 6.76 (br s, 0.4H), 6.74 (br s, 0.6H), 6.51-6.63 (br s, 2H), 5.96-6.00 (m, 2H), 5.68 (hr s, 0.4H), 5.58 (br s, 0.6H), 4.56-4.64 (m, 0.4H), 4.45-4.52 (m, H), 4.28-4.36 (m, H), 4.20-4.27 (m, 2H), 4.01-4.09 (m, 0.6H), 3.08-3.32 (m, 5H), 2.86-2.94 (m, IH), 2.69-2.76 (m, H), 2.31-2.37 (m, H), 1.96-2.22 (m, 4H), 1.89-1.96 (m, H), 1.30-1.80 (m, 2H), 1.10-1.16 (m, 4H), 1.04-1.09 (m, 2H); MS (ESI): mlz852.3 [M+H]'. (4)Synthesis of 4 with high purity was achieved following the method of Wilchek and Bayer with critical changes in workup procedures. 6-[5-(2-Oxo-hexahydro-thieno[3, 4-d]imidazol-4-yl)-pentanoylamino]-hexanoic acid (3; 0.31 g, 0.87 mmol) was completely dissolved in DMF (20 mL) with gentle warming.After cooling the 3 solution to room temperature without reprecipitation, DCC (0.34 g, 1.65 mmol) and pyridine (0.07 mL, 0.83 mmol) were added to the solution while stirring for 5 min, followed by the addition of NHS (0.16 g, 1.46 mmol) with continuous stirring for 18 h.The reaction product was filtered and concentrated under reduced pressure.The remaining solid was redissolved in 2-propanol (40 mL) with gentle heating, cooled down to room temperature, and reprecipitated at 4° C. overnight.The product was scraped out of the glassware, washed with cold 2-propanol, and air-dried to obtain 4 (0.22 g; 56.8percent) as a white solid. 1H NMR (400 MHz) (DMSO) delta: 7.77 (s, 1H, CONH), 6.45 (s, 1H, CONH), 6.38 (s, 1H, CONH), 4.31 (t, 1H, CHN), 4.15 (t, 1H, CHN), 3.11 (dd, 1H, CHS), 3.03 (t, 2H, CH2NH), 2.83 (s, 4H, CH2 of NHS), 2.79 (d, 1H, CHHS), 2.67 (t, 2H, CH2COOH), 2.60 (d, 1H, CHHS), 2.06 (t, 2H, CH2CO), 1.04-1.75 (m, 12H).13C NMR (100.67 MHz) (DMSO) delta: 171.90, 170.36, 169.03, 162.79, 61.13, 59.27, 55.53, 38.14, 35.32, 30.24, 28.74, 28.32, 28.14, 25.54, 25.41, 24.05. HRMS (ESI) calculated for C20H30N4O6S, [M+Na]+ 477.1784 (calcd.), 477.1784 (found).

A biotinylation reagent incorporating an aminocaproyl “spacer”.This can reduce the steric hindrance in binding avidin to some biotinylated compounds when used to biotinylate both the antibody and the carrier in the Protein Avidin-Biotin Capture System.This system avoids the direct interaction of a captured antibody with solid surfaces, such as plastics, which may reduce antigenicity.It has also been used toenhance the detection of DNA on nitrocellulose.Has also been used as a cell surface labelling reagent

Molecular Weight:454.5
XLogP3:0.1
Hydrogen Bond Donor Count:3
Hydrogen Bond Acceptor Count:7
Rotatable Bond Count:13
Exact Mass:454.18860586
Monoisotopic Mass:454.18860586
Topological Polar Surface Area:159
Heavy Atom Count:31
Complexity:702
Defined Atom Stereocenter Count:3
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes