Alternative to Piperidine in Fmoc Solid-Phase Synthesis

Hachmann, John; Lebl, Michal

doi:10.1021/cc050123l

Cited by 67 publications

(52 citation statements)

References 2 publications

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“…Diisopropylcarbodiimide was used for coupling (2 ϫ 1 h) and 20% 4-methylpiperidine (20,23) for Fmoc (N-(9-fluorenyl)methoxycarbonyl) group deprotection. Final deprotection and cleavage from the resin was performed by using Mixture K (82.5% TFA, 5% phenol, 5% H 2 O, 5% thioanisoee, 2.5% ethanedithioe) (24).…”

Section: Methodsmentioning

confidence: 99%

Targeting Host Cell Furin Proprotein Convertases as a Therapeutic Strategy against Bacterial Toxins and Viral Pathogens

Shiryaev

Remacle

Ratnikov

et al. 2007

Journal of Biological Chemistry

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100

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Pathogens or their toxins, including influenza virus, Pseudomonas, and anthrax toxins, require processing by host proprotein convertases (PCs) to enter host cells and to cause disease. Conversely, inhibiting PCs is likely to protect host cells from multiple furin-dependent, but otherwise unrelated, pathogens. To determine if this concept is correct, we designed specific nanomolar inhibitors of PCs modeled from the extended cleavage motif TPQRERRRKKR2GL of the avian influenza H5N1 hemagglutinin. We then confirmed the efficacy of the inhibitory peptides in vitro against the fluorescent peptide, anthrax protective antigen (PA83), and influenza hemagglutinin substrates and also in mice in vivo against two unrelated toxins, anthrax and Pseudomonas exotoxin. Peptides with Phe/Tyr at P1 were more selective for furin. Peptides with P1 Thr were potent against multiple PCs. Our strategy of basing the peptide sequence on a furin cleavage motif known for an avian flu virus shows the power of starting inhibitor design with a known substrate. Our results confirm that inhibiting furin-like PCs protects the host from the distinct furin-dependent infections and lay a foundation for novel, host cell-focused therapies against acute diseases.

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Section: Methodsmentioning

confidence: 99%

Targeting Host Cell Furin Proprotein Convertases as a Therapeutic Strategy against Bacterial Toxins and Viral Pathogens

Shiryaev

Remacle

Ratnikov

et al. 2007

Journal of Biological Chemistry

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100

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“…An additional 100 μl of DMF was added into the plate wells (beads sedimented) and the plate was centrifuged with a tilt of 6 degrees. A standard protocol was used for the synthesis to remove the Fmoc protecting group; 4-Methylpiperidine was used instead piperidine [16]. Individual Fmoc-protected amino acids (0.3M solution in 0.3M HOBt in DMF) were pipetted to the wells, and a solution of BOP (0.6M in DMF) and 1.2 M DIEA in DMF was delivered to each well.…”

Section: Methodsmentioning

confidence: 99%

Using support vector machine regression to model the retention of peptides in immobilized metal-affinity chromatography

Kermani

Kozlov

Melnyk

et al. 2007

Sensors and Actuators B: Chemical

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Retention of histidine-containing peptides in immobilized metal-affinity chromatography (IMAC) has been studied using several hundred model peptides. Retention in a Nickel column is primarily driven by the number of histidine residues; however, the amino acid composition of the peptide also plays a significant role. A regression model based on support vector machines was used to learn and subsequently predict the relationship between the amino acid composition and the retention time on a Nickel column. The model was predominantly governed by the count of the histidine residues, and the isoelectric point of the peptide.

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“…These amines trap very efficiently the DBF intermediate generating an adduct and, therefore, driving the deprotection step to completion [8][9][10][11]. The most used secondary amine is piperidine (pKa = 11.1, 25 °C), although one of the main reported problems with its use is the formation of aspartimide [12,13], which can be minimized by the use of other bases.…”

Section: Introductionmentioning

confidence: 99%

“…This fact implies that special permission is required for purchasing piperidine which, in certain institutions or countries, can cause administrative problems. In this regard, other cyclic secondary amines, 4-methyl piperidine, or piperazine (pKa = 10.78 and 9.73, 25 °C respectively) have been also proposed [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Deprotection Reagents in Fmoc Solid Phase Peptide Synthesis: Moving Away from Piperidine?

et al. 2016

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Abstract:The deprotection step is crucial in order to secure a good quality product in Fmoc solid phase peptide synthesis. 9-Fluorenylmethoxycarbonyl (Fmoc) removal is achieved by a two-step mechanism reaction favored by the use of cyclic secondary amines; however, the efficiency of the reaction could be affected by side reactions and by-product formation. Several aspects have to be taken into consideration when selecting a deprotection reagent: its physicochemical behavior, basicity (pKa) and polarity, concentration, and time of reaction, toxicity and disposability of residues and, finally, availability of reagents. This report presents a comparison of the performance of three strategies for deprotection using microwave-assisted Fmoc peptide synthesis. Four peptide sequences were synthesized using Rink amide resin with a Liberty Blue™ automated synthesizer and 4-methylpiperidine (4MP), piperidine (PP), and piperazine (PZ) as Fmoc removal reagents. In the first instance all three reagents behaved similarly. A detailed analysis showed a correlation between the hydrophobicity and size of the peptide with the yield and purity of the obtained product. The three reagents are interchangeable, and replacement of piperidine could be advantageous regarding toxicity and reagent handling.

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Alternative to Piperidine in Fmoc Solid-Phase Synthesis

Abstract: 4-Methylpiperidine is fully equivalent to piperidine in removal of Fmoc protecting group in solid phase synthesis. This reagent is not a controlled substance and its use does not require any paperwork.

Cited by 67 publications

References 2 publications

Targeting Host Cell Furin Proprotein Convertases as a Therapeutic Strategy against Bacterial Toxins and Viral Pathogens

Targeting Host Cell Furin Proprotein Convertases as a Therapeutic Strategy against Bacterial Toxins and Viral Pathogens

Using support vector machine regression to model the retention of peptides in immobilized metal-affinity chromatography

Deprotection Reagents in Fmoc Solid Phase Peptide Synthesis: Moving Away from Piperidine?

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