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Search for reports by entering the product batch number.
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Search for reports by entering the product batch number.
Batch number can be found on the product's label following the word 'Batch'.
Search for reports by entering the product batch number.
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CAS No. : | 84624-27-1 |
Formula : | C26H32N2O6 |
M.W : | 468.54 |
SMILES Code : | O=C(O)[C@H](CCCCNC(OCC1C2=C(C3=C1C=CC=C3)C=CC=C2)=O)NC(OC(C)(C)C)=O |
MDL No. : | MFCD00062032 |
InChI Key : | JYEVQYFWINBXJU-QFIPXVFZSA-N |
Pubchem ID : | 10983524 |
GHS Pictogram: | ![]() |
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 |
---|---|---|
General procedure: The second portion of the CCK4 peptide and the SubP peptide were modified on resin as follows to yield test lipidated peptides (1-SubP-COOH and 1-CCK4-Gly-COOH). Spacers (these are AA's used between the peg linker and the peptide of interest) were introduced on the peptides before pegylation (KGG for SubP and GG for CCK4). The free N-terminus of the peptide on resin was first pegylated with N-Fmoc-PEG8-propionic acid using standard HBTU coupling conditions. The N-Fmoc protecting group was removed by treatment with 10% piperidine in DMF (N,N-Dimethylformamide) for 5 min. Palmitic acid was subsequently coupled with the N-terminal free amine of the pegylated peptide. Peptides were cleaved from the resin using high HF conditions with minor modifications to the usual procedure. For the SubP peptide, longer times were used to ensure removal of Arg(Tos) protecting group (90% anhydrous HF/10% anisole at 0 C. for 2 h). For the CCK-4 peptides, 1,3-propanedithiol was used in the HF cleavage mixture to ensure deprotection of the formyl protecting group and prevent oxidation of methionine to its sulfoxide derivative: 85% anhydrous HF/10% anisole/5% PDT (1,3-propaneithiol) at 0 C. for 2 h) (Matsueda, 1982). Following cleavage from resin, peptides were precipitated with cold Et2O. Unmodified peptides were extracted using 10% AcOH in water and the lipidated peptides were extracted using 10% AcOH in H2O followed by 10% AcOH in 50% EtOH/H2O. Crude peptides were purified by RP-HPLC [Vydac C18, 10 m, 22 mm×250 mm]. The purities of the peptides were assessed by analytical RP-HPLC [Vydac C18, 5 m, 4 mm×250 mm]. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Peptides illustrated in FIG. 9 were synthesized using the in-situ neutralization protocol for t-Boc chemistry (Schnolzer et al., In situ neutralization in Boc-chemistry solid phase peptide synthesis. Rapid, high yield assembly of difficult sequences, 1992, International journal of peptide and protein research 40:180-193) on PAM resin on a 0.5 mmol scale. Amino acids were used with the following side chain protecting groups: Arg(Tos), Asp(OBzl), Gln(Xan), Lys(Fmoc), Lys(2-Cl-Z) and Trp(For). Peptide coupling reactions were carried out with a 4-fold excess (2.0 mmol) of activated amino acid for at least 15 min. The t-Boc protecting group on the N-terminus was removed using trifluoroacetic acid (TFA). The PAM resin from the CCK4 peptide synthesis was split into two equal portions. One portion of the resin was used for synthesizing non-lipidated peptides. The CCK4 (s-CCK-Gly-COOH) peptide was left unmodified at the N-terminus. This peptide served as positive control for the lipidated counterparts. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
EXAMPLE 11 Synthesis of Glucagon Lactams (0299) 285 mg (0.2 mmole) methoxybenzhydrylamine resin (Midwest Biotech) was added to a 60 mL reaction vessels and the following sequence was assembled on a modified Applied Biosystems 430A peptide synthesizer using Boc DEPBT-activated single couplings. HSQGTFTSDYSKYLDERRAQDFVQWLMNT-NH2 (12-16 Lactam; SEQ ID NO: 12) (0300) The following side chain protecting groups were used: Arg(Tos), Asp(OcHx), Asn(Xan), Glu(OFm), His(BOM), Lys(Fmoc), Ser(Bzl), Thr(Bzl), Trp(CHO), Tyr(Br-Z). Lys(Cl-Z) was used at position 12 if lactams were constructed from 16-20, 20-24, or 24-28. The completed peptidyl resin was treated with 20% piperidine/dimethylformamide for one hour with rotation to remove the Trp formyl group as well as the Fmoc and OFm protection from Lys12 and Glu16. Upon confirmation of removal by a positive ninhydrin test, the resin was washed with dimethylformamide, followed by dichloromethane and than again with dimethylformamide. The resin was treated with 520 mg (1 mmole) Benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP) in dimethylformamide and diisopropylethylamine (DIEA). The reaction proceeded for 8-10 hours and the cyclization was confirmed by a negative ninhydrin reaction. The resin was washed with dimethylformamide, followed by dichloromethane and subsequently treated with trifluoroacetic acid for 10 minutes. The removal of the Boc group was confirmed by a positive ninhydrin reaction. The resin was washed with dimethylformamide and dichloromethane and dried before being transferred to a hydrofluoric acid (HF) reaction vessel. 500 muL p-cresol was added along with a magnetic stir bar. The vessel was attached to the HF apparatus (Peninsula Labs), cooled in a dry ice/methanol bath, evacuated, and approximately 10 mL of liquid hydrofluoric acid was condensed into the vessel. The reaction was stirred for 1 hour in an ice bath and the HF was subsequently removed in vacuo. The residue was suspended in ethyl ether; the solids were filtered, washed with ether, and the peptide was solubilized with 150 mL 20% acetonitrile/1% acetic acid. (0301) An analytical HPLC analysis of the crude solubilized peptide was conducted under the following conditions [4.6 X 30 mm Xterra C8, 1.50 mL/min, 220 nm, A buffer 0.1% TFA/10% ACN, B buffer 0.1% TFA/100% ACN, gradient 5-95%B over 15 minutes]. The extract was diluted twofold with water and loaded onto a 2.2 X 25 cm Vydac C4 preparative reverse phase column and eluted using an acetonitrile gradient on a Waters HPLC system (A buffer of 0.1% TFA/10% ACN, B buffer of 0.1% TFA/10% CAN and a gradient of 0-100% B over 120 minutes at a flow of 15.00 ml/min. HPLC analysis of the purified peptide demonstrated greater than 95% purity and electrospray ionization mass spectral analysis confirmed a mass of 3506 Da for the 12-16 lactam. Lactams from 16-20, 20-24, and 24-28 were prepared similarly. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
The title peptide was synthesized on an Applied Biosystems (Foster City, Calif.) model 430A peptide synthesizer which was modified to do accelerated Boc-chemistry solid phase peptide synthesis. See Schnolzer, et al., Int. J. Peptide Protein Res., 40:180 (1992). 4-methylbenzhydrylamine (MBHA) resin (Peninsula, Belmont, Calif.) with the substitution of 0.91 mmol/g was used. The Boc amino acids (Novabiochem, San Diego, Calif. and Chem-Impex, Wood Dale, Ill.) used were: Boc-Cha-OH, Boc-Asp(OFm)-OH, Boc-His(DNP)-OH, Boc-D-Phe-OH, Boc-Arg(Tos)-OH, <strong>[47355-10-2]<strong>[47355-10-2]Boc-Trp(For)</strong>-OH</strong>, Boc-Gaba-OH, and Boc-Lys(Fmoc)-OH. The synthesis was carried out on a 0.20 mmol scale. The Boc groups were removed by treatment with 100% TFA for 2×1 minute. Boc amino acids (2.5 mmol) were pre-activated with HBTU (2.0 mmol) and DIEA (1.0 mL) in 4 mL of DMF and were coupled without prior neutralization of the peptide-resin TFA salt. Coupling times were 5 minutes. (0223) At the end of the assembly of Boc-Asp(OFm)-His(DNP)-D-Phe-Arg(Tos)-Trp(For)-Gaba-Lys(Fmoc)-MBHA, the peptide-resin was transferred into a reaction vessel on a shaker. The resin was treated twice with 25% piperidine in DMF for 15 minutes per session, washed with DMF, and shaken with bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBrOP) (6 eq, 0.3 mmol), DIEA (1 mL), and 4-(dimethylamino)pyridine (DMAP) (24 mg) in DMF (2 mL) for 12 hours. After washing with DMF, the resin was treated twice with 100% TFA for 2 minutes per treatment, washed with DMF and DCM, and then dried under reduced pressure. One fourth of the peptide-resin (0.05 mmol) was used for the next coupling with Boc-Cha-OH (10 eq, 0.5 mmol) in the presence of HBTU (9 eq, 0.45 mmol) and DIEA (0.25 mL) in DMF for 10 minutes. After the deprotection with 100% TFA in two sessions lasting approximately 2 minutes each, the peptide-resin was then washed with DMF. The final capping step was done by shaking the resin with acetic anhydride (40 eq, 2.0 mmol) and DIEA (20 eq, 1.0 mmol) in DMF for 1 hour. After washing with DMF, the resin was treated twice with a solution of 20% mercaptoethanol/10% DIEA in DMF, each treatment lasting approximately 30 minutes, to remove the DNP group on the Histidine side chain. The formyl group on the side chain of Tryptophan was removed by shaking with a solution of 15% ethanolamine/15% water/70% DMF twice for 30 minutes per shaking. The peptide-resin was washed with DMF and DCM and dried under reduced pressure. The final cleavage was done by stirring the peptide-resin in 10 mL of HF containing 1 mL of anisole and dithiothreitol (30 mg) at 0 C. for 75 minutes. HF was removed by a flow of nitrogen. The residue was washed with ether (6×10 mL) and extracted with 4N HOAc (6×10 mL). (0224) The peptide mixture in the aqueous extract was purified on reverse-phase preparative high pressure liquid chromatography (HPLC) using a reverse phase VYDAC C18 column (Nest Group, Southborough, Mass.). The column was eluted with a linear gradient (10% to 50% of solution B over 40 minutes) at a flow rate of 10 mL/minute (Solution A=water containing 0.1% TFA; Solution B=acetonitrile containing 0.1% of TFA). Fractions were collected and checked on analytical HPLC. Those containing pure product were combined and lyophilized to dryness. 5.1 mg of a white solid was obtained. Yield was 8.9%. Purity was 94.5% based on analytical HPLC analysis. Electro-spray mass spectrometer (MS(ES))S analysis gave the molecular weight at 1148.5 (in agreement with the calculated molecular weight of 1148.3). (0225) Other peptides of the invention can be prepared by a person of ordinary skill in the art using synthetic procedures analogous to those disclosed generally hereinabove and/or to those disclosed specifically in the foregoing examples, as were the compounds depicted in Tables 1A and 1B. (0226) Other peptides of the invention can be prepared by a person of ordinary skill in the art using synthetic procedures analogous to those disclosed generally hereinabove and/or to those disclosed specifically in the foregoing examples, as were the compounds depicted in Tables 1A and 1B. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
The procedure for the synthesis of this group of compounds of the present invention is discussed below. The peptide H-Lys(hArg)-Pro-Dab-Arg-COOH was chosen as the example: 160 mg of pre-loaded Fmoc-L-Arg(Pbf) WANG resin was mixed in 4 ml of the anhydrous N,N- dimethylformamide (DMF) for four hours. After that time, the resin was filtered off and then mixed in 6 ml of 20percent (v/v) piperidine in DMF for 5 minutes, then the resin was filtered off and a fresh aliquot of 20percent piperidine in DMF was added (6 ml) and stirred for another 20 minutes. The next step of the synthesis was the alternately washing of the resin with 6 mL of dry DMF and 6 ml of isopropanol (IPA) until neutral pH, usually three times for one minute each. At the end of this step of the synthesis, the resin was washed three times (for one minute) with a fresh portion of anhydrous DMF. After the wash sequence, for monitoring of the completion of the Fmoc cleavage, the colorimetric assay was made. Usually, for primary amines, this is the Kaiser test. For this purpose a few resin beads were placed in small test tube and few drops (equal volumes) of the three solutions [(A): 5 g of ninhydrin in 100 ml ethanol; (B): 80 g phenol in 20 ml of ethanol; (C) 2 ml 0.001 M aqueous KCN in 98 ml pyridine] were added. The test tube was placed in a water bath and heated for 5 minutes at 100°C. A positive Kaiser test result (deep blue color of resin and solution) was obtained which allowed us to proceed to the next step wherein 88 mg (0.2 mmol) of Fmoc-Dab(Boc)-OH, 64 mg (0.2 mmol) TBTU and 51 mL (0.3 mmol) DIPEA were dissolved in 5 ml of anhydrous DMF - to form the preactivation mixture. The preactivation mixture was stirred for 10 minutes and added to the resin and mixed for another 10 minutes, then additional 51 mL (0.3 mmol) of DIPEA was added and mixing was continued for another four hours. After this time the resin was filtered off and washed four times (1 minute) with a fresh portion (6 ml) of anhydrous DMF, and then again the Kaiser test was performed with negative result (which means the completion of the coupling reaction). Next, the resin was mixed in 6 ml of 20percent (v/v) piperidine in DMF for 5 minutes, then the resin was filtered off and fresh aliquot of 20percent piperidine in DMF was added (6 ml) and stirred for another 20 minutes. The further step of the synthesis was the alternately washing of the resin with 6 mL of dry DMF and 6 ml of isopropanol (IPA) until neutral pH, usually three times (DMF and IPA) for one minute each. At the end of this step of the synthesis, the resin was washed three times (for one minute) with a fresh portion of anhydrous DMF. After the wash sequence, for monitoring of the completion of the Fmoc cleavage, the Kaiser test was made. Also in this case, the positive result of Kaiser test was obtained which allowed to proceed to the next step wherein 67 mg (0.2 mmol) of Fmoc-Pro-OH, 64 mg (0.2 mmol) of TBTU and 51 mL (0.3 mmol) of DIPEA were dissolved in 5 ml of anhydrous DMF - to form the preactivation mixture. The preactivation mixture was stirred for 10 minutes and added to the resin and mixed for another 10 minutes, then additional 51 mL (0.3 mmol) of DIPEA was added and mixing was continued for another four hours. After this time, the resin was filtered off and washed four times (1 minute) with a fresh portion (6 ml) of anhydrous DMF, and then again the Kaiser test was performed. The result of Kaiser test was negative, which means the completion of the coupling reaction. Next, the resin was mixed in 6 ml of 20percent (v/v) piperidine in DMF for 5 minutes, then the resin was filtered off and a fresh aliquot of 20percent piperidine in DMF was added (6 ml) and stirred for another 20 minutes. The further step of the synthesis was the alternately washing of the resin with 6 mL of dry DMF and 6 ml of isopropanol (IPA) until neutral pH, usually three times (DMF and IPA) for one minute each. At the end of this step of the synthesis, the resin was washed three times (for one minute) with a fresh portion of anhydrous DMF. For monitoring of the completion of the Fmoc cleavage from proline residue the acetaldehyde/chloranil test was performed. For this purpose a few resin beads were placed in small test tube and few drops (equal volumes) of the two solutions [(A): 2percent acetaldehyde in DMF; (B): 2percent chloranil in DMF] were added. After a short mixing the mixture was left at room temperature for 5 minutes. After that time, the dark blue color of beads of resin indicated the completion of the Fmoc cleavage. In the next step of the synthesis, the preactivation mixture consist of the 94 mg (0.2 mmol) of Boc-Lys(Fmoc)-OH, 64 mg (0.2 mmol) of TBTU,51 mL of DIPEA and dissolved in 5 mL of dry DMF was stirred for 10 minutes and added to the resin and mixed for another 10 minutes, then additional 51 mL (0.3 mmol) of DIPEA was added and mixing was continued for another four hours. After this time, the resin was filtered off... |
Tags: Boc-Lys(Fmoc)-OH | Carboxylic Acids | Carbamates | Organic Building Blocks | 84624-27-1
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H401 | Toxic to aquatic life |
H402 | Harmful to aquatic life |
H410 | Very toxic to aquatic life with long-lasting effects |
H411 | Toxic to aquatic life with long-lasting effects |
H412 | Harmful to aquatic life with long-lasting effects |
H413 | May cause long-lasting harmful effects to aquatic life |
H420 | Harms public health and the environment by destroying ozone in the upper atmosphere |
Sorry,this product has been discontinued.
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The concentration of the dissolution solution you need to prepare is mg/mL