The tfa filtrate could be used in two ways, depending on the size of the peptide. Myers protective groups siliconbased protection of the. Protecting groups of hydroxyl groups consider the stability and effect of anomeric group. First the resin is separated from tfa by filtration through a mediumporosity fritted glass funnel and the resin is washed with 1 ml of tfa. I have seen some procedures of pmb deprotection of amine using trifluro acetic acid. Mild, selective deprotection of pmb ethers with triflic. The deprotection cleavage of the boc group is realized using a 50% solution of tfa in dcm.
Received 29 june 2011 revised 16 august 2011 accepted 18 august 2011 available online 25. Protection and deprotection of functional groups in. The reaction mechanism is unlike the s n 1 path in carbon chemistry as silyl cations are unstable. Specifically, we disclose onresin deblocking using trifluoroacetic acid tfatriisopropylsilane tis in etoacmecn used in a synthesis of a melanocortin receptor agonist comprising i 1d green spps ii 2d green spps by an onresin tfatis in etoacmecn deprotection of lysmtt and aspo2phipr followed by a lactamization using pybopdiea. A eight residue peptide containing phosphotyrosine was synthesized with boctyropo 3 bzl 2.
A practical, catalytic and selective deprotection of a boc group in n,n. The purity of each fraction was verified by analytical hplc and malditof and showed that the peptides were 94% pure. An interesting example of selective deprotection of the tertbutyl esters in the presence of nboc protecting groups of several amino acids using ceriumiii chloride as a lewis acid is reported. Deprotection akin to protection, the deprotection proceeds through pentacoodinated intermediates. Aggregation, racemization and side reactions in peptide. The phosphoamino acid was double coupled, the peptide cleaved and extracted using the tfmsa tfa procedure outlined in materials and methods. This protocol proceeds from the premise that dotatristbe has been conjugated to a molecule of interest e. Try it first, as it can take longer then you think also you need to track your products.
Deprotection of s10undecenyl thioacetate figure 2 dissolve s10undecenyl thioacetate 1 2. Consider the reactivity of different hydroxyl groups. I know boc can easily be deprotected using strong acid like tfa and hcl, but final product is salt of tfa or hcl, my question is that, how we can avoid formation of slat and obtained free amine group. The two sets of peptides were deprotected with either 55% tfadcm for 30 min or 100% tfa for 5 min. Consider the solubility of starting material the choice of solvent. Trifluoroacetic acid tfa and bromide, formed mainly by maternal metabolism of halothane, accumulated in fetus and amniotic fluid with time, and reached plateau levels in amniotic fluid between 4 and 24 hr. Comparison of 55% tfach2cl2 and 100% tfa for boc group. The co2 gas that forms during the reaction should be allowed to escape. Cleavage, deprotection, and isolation of peptides after fmoc.
Lewis acids such as znbr 2, ticl 4, me 2bbr 2 meoch 2ch 2och 2cl, ch 2cl 2, ipr 2etn stable to base and mild acid roh roch2och2ch2ome. The org layer was separated, dried mgso4, and concentrated in vacuo to provide the product as a brown solid. The pmb group was easily and selectively removed in 6898% yield using tfa in dcm. A necessary evil 3 note, however, that each protecting group incorporated in a multistep synthesis increases the synthesis by two nonproductive steps reducing the overall yield and efficiency of the synthesis. Ive done many insitu deprotection in dcm after a coupling. The workup of the deprotection reaction consists of precipitation of the peptide product from tfa. The usefulness of adding scavengers to trifluoroacetic acid in deblocking reactions is due to the removal of t. Rapid deprotection of n boc amines by tfa combined with. Protecting groups hydroxyl protecting groups 4 ethers 3. A novel series of 3,6bis4hdroxyphenyl1,2,4,5tetrazine alkanoate esters were.
Peptides are chemically synthesized by the condensation reaction of the carboxyl group of one amino acid to the amino group of another. The phosphoamino acid was double coupled, the peptide cleaved and extracted using the tfmsatfa procedure outlined in materials and methods. I need the product to be a tfa salt and not an hcl salt so i need to use tfa. The benzyl ester linkage between the cterminal amino acid and the polystyrene can be partially cleaved in the deprotection step and important losses can occur during the construction of a long peptide chain 3. General procedure for the deprotection of the bocgroup. Protection of the amine can also be accomplished in acetonitrile solution using 4. If the beads turn orange immediately, continue shaking for another 30 minutes and retest. Choose from 500 different sets of tfa chapter 14 flashcards on quizlet. Boc falls off fairly easily in some cases, but in your case you are better to use tfa. A practical, catalytic and selective deprotection of a boc.
New tfafree cleavage and final deprotection in fmoc solid. Treatment of peptides containing sacm protecting group with iodine results in simultaneous removal of the sulfhydryl protecting group and disulfide formation. In organic chemistry, peptide synthesis is the production of peptides, compounds where multiple amino acids are linked via amide bonds, also known as peptide bonds. The sacetamidomethyl acm or trityl trt protecting groups are widely used in the chemical synthesis of peptides that contain one or more disulfide bonds. This cton synthesis is opposite from protein biosynthesis, during which the nterminus of the incoming amino acid is linked to the cterminus of the protein chain ntoc. Tfa equal amount as the solvent was added and the solution was allowed to warm to room temperature. A selective, catalytic and practical method for removing a boc group from several n,n. Protic acids such as trifluoroacetic acid tfa, commonly used for. After stirring at room temperature until starting material was. The driving force of fluoridebased deprotection is the formation of sif bond, which is about 30 kcalmol stronger than sio bond. Eventually i pass ammonia gas to remove tfasalt but unfortunate atr is not showing any primary amine nh2 peak yet.
I have done bocdeprotection by tfa, and reaction was monitored by tlc. Tentagel 12 1987 rink and others introduction of various tfalabile linkers for the fmoctbu spps of peptide amides. What happens to the tbutyl cation in the tfa deprotection of. Even though the cleavage of the tbutylcarbamate boc group can be achieved at solvent reflux temperature, the deprotection process was significantly accelerated under microwaveassisted conditions. Thus, deprotection using tfa is now the final step to be accomplished. How to do workup of a boc deprotection reaction by tfa. Decarboxylation of the carbamic acid results in the free amine. Protecting group pg is a small molecule, to mask temporarily the a specific functional group of a molecule from undergoing reaction, allowing the rest of the functional groups present in the molecule to react without affecting the original reactivity and leave from the host molecule without affecting the rest of the functional groups. However, the excess iodine needs to be quenched or adsorbed as quickly as. The thermolytic deprotection of nboc compounds was accomplished using tfe 2,2,2trifluoroethanol or hfip hexafluoroisopropanol as solvents. Read 1 answer by scientists with 2 recommendations from their colleagues to the. Suspend the resin in 50% vv tfadichloromethane dcm, using 1 ml of tfadcm per gram of resin.
Another attempt of deprotection of 6a,b was performed by hydrogenolysis in thf using pdc 5 h, rt, which gave the deprotected derivatives 1a,b with excellent yields xzo. Selective deprotection of lysmtt cleavage with tfatisdcm. This procedure is suitable for the parallel deprotection of nboc amines. Isobutene reacts with trifluoroacetic acid and yields t.
Catalog technical deprotection pdf, forms catalog technical deprotection pdf, reference technical deprotection pdf, forms technical deprotection pdf. The purity of the peptides obtained when deprotecting with 55% tfa dcm was, on average, 9% higher than with 100% tfa. Method g with bu 4nf in thf was found to be a very good alternative 6a. If tbutyltrifluoroacetate is the byproduct, then the catalyst is consumed. Protecting groups in organic synthesis1 ready protecting groups are a sad fact of synthetic chemistry they are usually needed, but rarely desired many syntheses have stalled because of trouble putting on or removing protecting groups 4 basic questions to address when choosing a p. A theoretical study rationalizing the results with several metals i.
The boc group can be added to the amine under aqueous conditions using di tert butyl dicarbonate in the presence of a base such as sodium bicarbonate. Any suggestion on boc deprotection without using acid. Loss of the tertbutyl cation results in a carbamic acid. Novel practical deprotection of nboc compounds using. New tfafree cleavage and final deprotection in fmoc solidphase. Myers protective groups siliconbased protection of the hydroxyl group chem 115 general reference. In the initial step, protonation is more likely on the co oxygen. Read here what the tfa file is, and what application you need to open or convert it. All synthetic steps, other than the removal of the boc group, were performed simultaneously under identical experimental conditions. Study on the process of synthesizing methylenedinitramine by a novel method. Trimethylsilyl tms triethylsilyl tes triisopropylsilyl tips dimethylisopropylsilyl.
The tertbutyl cation will either be quenched by a suitable trapping agent, deprotonate to form isobutylene gas, or polymerize to form isobutylene oligomers. Tfa infused intravenously to the mother reached higher levels in amniotic fluid after long survival times, than in maternal plasma. Deprotection reaction an overview sciencedirect topics. Protonation of amine under the acidic conditions provides the pdt as the tfa salt. Its known that hexanes will form an azeotrope with acetic acid, so you could try redissolving in hexanes and spinning it down a few times.
Tunable acidsensitive ester protecting groups in oligosaccharide. Tentagel 12 1987 rink and others introduction of various tfa labile linkers for the fmoctbu spps of peptide amides. Protecting groups in solidphase organic synthesis acs. Cleavage and deprotection is one of the most crucial steps in. The purity of the peptides obtained when deprotecting with 55% tfadcm was, on average, 9% higher than with 100% tfa. This protocol outlines the use of tfa to remove tertbutyl ester groups from dotatristbe. Mild, selective deprotection of pmb ethers with triflic acid. More important, compounds that are sensitive to typical acidic conditions tfa and hcl such as 5d and 23 gave the corresponding deprotected product in.
Bocprotected amino groups organic chemistry portal. The tertbutyloxycarbonyl protecting group or tertbutoxycarbonyl protecting group boc group is a protecting group used in organic synthesis the boc group can be added to the amine under aqueous conditions using ditertbutyl dicarbonate in the presence of a base such as sodium bicarbonate. Deprotection of boc using taf to obtained free amine group. A method for removal of nboc protecting groups from. The boc group has also been cleaved with conditions that do not use tfa. The two sets of peptides were deprotected with either 55% tfa dcm for 30 min or 100% tfa for 5 min.
Deprotection of glutamic acid residues during hf cleavage can result in the formation of an acylium ion from the hf protonation and dehydration of the unprotected carboxyl moiety. How to confirm boc deprotection by tfa researchgate. Fmoc protecting group, which is stable to tfa deprotection, must be removed by a 20% solution of piperidine in dmf. Protection and deprotection of functional groups in organic. In bocbased synthesis, diketopiperazine formation can be suppressed by utilizing in situ neutralization protocols. A simple protocol for the deprotection of 1,3dithianes and 1,3dithiolanes showed tolerance for a number of phenol and amino protecting groups using 30% aqueous hydrogen peroxide activated by iodine catalyst 5 mol% in water in the presence of sodium dodecyl sulfate sds under essentially neutral conditions without any detectable overoxidation.
After the reaction of phenylalanine boc deprotection used tfa dcm1. Though it is difficult to see nh2 nmr peak, in ir primary amines rnh2 display. Mild, selective deprotection of pmb ethers with tri. The acylium ion can cyclize to produce a pyroglutamine residue or it can react with scavengers such as anisole to form an aryl ketone. After the reaction of phenylalanine boc deprotection used tfadcm1.
Albr3, etsh phse ph2p me3sii o o obz ome o o obz oh albr 3, etsh tl 1987, 28, 3659 methoxymethyl ether mom roh roch2ome stable to base. Reaction times of 36 h and overall yields of 7898% were achieved with the pmb group in place compared to no reaction without this protecting group. Br removal tbaf, 35% tfa pdpph 3 4, scavengers photolytic cleavage 320 nm stable basic conditions basic and acidic conditions acidic and mild basic conditions orthogonal fmoc boc, fmoc, zfmoc, boc 10. Peptide synthesis most often occurs by coupling the carboxyl group of the incoming amino acid to the nterminus of the growing peptide chain. Learn tfa chapter 14 with free interactive flashcards. New tfafree cleavage and final deprotection in fmoc solidphase peptide synthesis. If the fmoctbu protection strategy is utilized, performing the synthesis on 2chlorotrityl chloride resin is preferred when proline, pipecolic acid or tic is one of the first two amino acids. For an acetylated map peptide, the removal of the fmoc protecting group is followed by capping with acetic anhydride to give an acetylated map peptide. The respective peptide was dissolved in ch2cl 2 or in ch 2 cl 2.
The freebase amines are then obtained by scavenging the crude reaction mixture with the basic amberlyst a21 ionexchange resin. Kinetic studies show comparable reaction rates with thiophenol as scavenger. Scheme 1 efficient global removal of nap ethers in dodecyl maltoside 1 by tfa in. During cleavage of peptide after solid phase synthesis. The solvent was removed in vacuo and the crude was dissolved in 10% ipachloroform, washed with sat aq nahco3, brine, dried, and concentrated. The crude was purified by semipreparative hplcms using a linear gradient from 25 to 30% of mecn in 12 min of mecn containing 0.
In addition, removal of the nde group could be followed visually and monitored. Ntertbutyloxycarbonyl boc groups can be removed from peptides supported on rinks amide resin by treatment with trimethylsilyl triflate2,6lutidine. Excess 1 n naoh was added and the mixture was stirred vigorously for 15 min. The deprotection of nboc amines was rapidly accomplished using 5 equivalents of tfa in methylene chloride in a focused microwave instrument with irradiati. The process is clean, not needing a purification step. Intermediate intermediate diamine 7 was obtained as a yellow oil 22 mg, quant. A novel method for cleaving from resin and removing acidlabile protecting groups for the fmoc solidphase peptide synthesis is described. A pmethoxybenzyl pmb protectiondeprotection approach. Tfa 9 1985 houghten and others simultaneous parallel peptide synthesis, synthesis of peptide libraries tbags, pins, etc.