Purity | Size | Price | VIP Price | USA Stock *0-1 Day | Global Stock *5-7 Days | Quantity | |||||
{[ item.p_purity ]} | {[ item.pr_size ]} |
{[ getRatePrice(item.pr_usd, 1,1) ]} {[ getRatePrice(item.pr_usd,item.pr_rate,item.mem_rate) ]} |
{[ getRatePrice(item.pr_usd, 1,1) ]} | Inquiry {[ getRatePrice(item.pr_usd,item.pr_rate,item.mem_rate) ]} {[ getRatePrice(item.pr_usd,1,item.mem_rate) ]} | {[ item.pr_usastock ]} | Inquiry - | {[ item.pr_chinastock ]} | Inquiry - |
* Storage: {[proInfo.prStorage]}
CAS No. : | 86060-81-3 | MDL No. : | MFCD00038769 |
Formula : | C21H22N2O5S | Boiling Point : | - |
Linear Structure Formula : | - | InChI Key : | CSMYOORPUGPKAP-IBGZPJMESA-N |
M.W : | 414.47 | Pubchem ID : | 128799 |
Synonyms : |
|
Num. heavy atoms : | 29 |
Num. arom. heavy atoms : | 12 |
Fraction Csp3 : | 0.29 |
Num. rotatable bonds : | 11 |
Num. H-bond acceptors : | 5.0 |
Num. H-bond donors : | 3.0 |
Molar Refractivity : | 110.19 |
TPSA : | 130.03 Ų |
GI absorption : | High |
BBB permeant : | No |
P-gp substrate : | Yes |
CYP1A2 inhibitor : | No |
CYP2C19 inhibitor : | No |
CYP2C9 inhibitor : | Yes |
CYP2D6 inhibitor : | No |
CYP3A4 inhibitor : | Yes |
Log Kp (skin permeation) : | -6.88 cm/s |
Log Po/w (iLOGP) : | 1.92 |
Log Po/w (XLOGP3) : | 2.75 |
Log Po/w (WLOGP) : | 2.81 |
Log Po/w (MLOGP) : | 1.72 |
Log Po/w (SILICOS-IT) : | 2.63 |
Consensus Log Po/w : | 2.36 |
Lipinski : | 0.0 |
Ghose : | None |
Veber : | 1.0 |
Egan : | 0.0 |
Muegge : | 0.0 |
Bioavailability Score : | 0.56 |
Log S (ESOL) : | -3.72 |
Solubility : | 0.0785 mg/ml ; 0.000189 mol/l |
Class : | Soluble |
Log S (Ali) : | -5.14 |
Solubility : | 0.00303 mg/ml ; 0.00000732 mol/l |
Class : | Moderately soluble |
Log S (SILICOS-IT) : | -5.74 |
Solubility : | 0.000752 mg/ml ; 0.00000182 mol/l |
Class : | Moderately soluble |
PAINS : | 0.0 alert |
Brenk : | 1.0 alert |
Leadlikeness : | 2.0 |
Synthetic accessibility : | 4.45 |
Signal Word: | Warning | Class: | N/A |
Precautionary Statements: | P261-P305+P351+P338 | UN#: | N/A |
Hazard Statements: | H315-H319-H335 | Packing Group: | N/A |
GHS Pictogram: |
* 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 benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 2h; | All the peptides were assembled manually on Rink amideMBHA resin (0.15g, 0.4 mmol/g) using the conventional Fmoc chemistry in a glass reaction vessel. The coupling reactions were carried out by adding a pre-activated solution of N-alpha-Fmoc-protected amino acid (3 equiv), HOBT (3 equiv), HBTU (3 equiv), and DIEA (6 equiv) in anhydrous DMF (10 mL/g resin) into the resin (1 equiv) and swirling for 2 h. The progress of the coupling was monitored by Kaiser test. The Fmoc protecting groups were removed by two 10 min treatments with a piperidine/DMF solution (20percent). The resin was washed by swirling in methanol (1 min, 2X), followed by DMF (1 min, 6X). The resin bound di-Cys(Acm) peptide was swirled with TI(F3CCOO)3 (2 equiv) in DMF to form the disulfide peptide, washed with DMF (1 min, 2X), and finally EPO <DP n="40"/>deblocked with 20percent piperidine in DMF. All the crude products were purified by semi-preparative HPLC and identified by both ES-MS and analytical HPLC. They showed the corresponding [MH]+, [MH2]2+ and even triple [MH3]3* peaks in their ES-MS spectra. A mixture of Cypate (211.5 mg, 0.3 mmol), DIC (38.0 mg,0.3 mmol), HOBT (41.0 mg, 0.3 mmol) and the resin bound disulfide RGD peptide (-0.06 mmol) in anhydrous DMF (3mL) was swirled for 8 h. The resin was filtered, washed with DMF (1 min, 3X), methanol (1 min, 2X), and dried under vacuum. The resin was cleaved with TFA (3 mL, 1.5 h, 5X). The TFA filtrate was concentrated and added to cold MBTE (10 mL). The solid precipitate was collected by filtration, washed with petroleum ether, and purified by semi-preparative HPLC to afford the desired products. The protected RGD peptide sequence, i.e. Fmoc-Cys(Acm)-Arg(Pbf)-Gly-Asp(OBut)-XAA-Cys(Acm) [SEQ ID NO:16], was first assembled on Rink amide MBHA resin (1 equiv) using the conventional Fmoc chemistry (Scheme 1). It was swirled with TI(F3CCOOH)3 (2 equiv) in DMF for 2 h to form the cyclic disulfide peptide, i.e., Fmoc-cyclo[Cys-Arg(Pbf)-Gly- Asp(OBut)-XAA-Cys] [SEQ ID NO:17] The conjugation of cypate with peptide usually afforded the monomeric and dimeric conjugates simultaneously. To increase the yield of monomeric product, we used 5 equivalent Cypate with 3.5 equivalent DIC and HOBT. Finally, TFA cleavage afforded the desired product, i.e., Cypate-cyclo(Cys-Arg-Gly-Asp-XAA-Cys)-NH2 [SEQ ID NO: 18]. The other cyclic disulfide RGD peptide analogs (Table 2) were synthesized using the same protocol. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
The procedure described below is for the synthesis of Octreotate. The amino acid sequence of Octreotate is: D-Phe-Cys'-Tyr-D-Trp-Lys-Thr-Cys'-Thr (SEQ ID NO:1), wherein Cys' indicates the presence of an intramolecular disulfide bond between two cysteine amino acids. Other peptides of this invention were prepared by a similar procedure with slight modifications in some cases. [0080] The octapeptide was prepared by an automated fluorenylmethoxycarbonyl (Fmoc) solid phase peptide synthesis using a commercial peptide synthesizer from Applied Biosystems (Model 432A SYNERGY Peptide Synthesizer). The first peptide cartridge contained Wang resin pre-loaded with Fmoc-Thr on 25-mumole scale. Subsequent cartridges contained Fmoc-protected amino acids with side chain protecting groups for the following amino acids: Cys(Acm), Thr(t-Bu), Lys(Boc), Trp(Boc) and Tyr(t-Bu). The amino acid cartridges were placed on the peptide synthesizer and the product was synthesized from the C- to the N-terminal position. The coupling reaction was carried out with 75 mumoles of the protected amino acids in the presence of 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU)/N-hydroxybenzotriazole (HOBt). The Fmoc protecting group was removed with 20percent piperidine in dimethylformamide. After the synthesis was complete, the thiol group was cyclized with thallium trifluoroacetate and the product was cleaved from the solid support with a cleavage mixture containing trifluoroacetic acid (85percent):water (5percent):phenol (5percent):thioanisole (5percent) for 6 hours. The peptide was precipitated with t-butyl methyl ether and lyophilized with water:acetonitrile (2:3) mixture. The peptide was purified by HPLC and analyzed with LC/MS. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS procedure starting from 2-Cl-Trt-chloride resin. The first amino acid (Fmoc-Gly) is loaded on the resin as described in previous examples to obtain a loading of about 0.7 mmol/g of amino acid/resin. After washing of the resin and removal of the Fmoc group by treatment with piperidine/DMF, the second amino acid (Fmoc-Orn(Boc)) is introduced to continue sequence elongation. Fmoc protected amino acids are activated in situ using TBTU/HOBt and subsequently coupled to the resin over about 50 minutes. Diisopropylethylamine or collidine is used during coupling as an organic base. Completion of the coupling is indicated by ninhydrin test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20% piperidine in DMF for 20 min. These steps are repeated each time with another amino acid according to the peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Cys(Acm), Thr(tBu), Asn(Trt), and Orn(Boc). Three equivalents of the activated amino acids are used in the coupling reactions. At the end of the synthesis, the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS procedure starting from 2-Cl-Trt-chloride resin. The first amino acid (Fmoc-Trp) is loaded on the resin as described in previous examples to obtain a loading of about 0.7 mmol/g of amino acid/resin. After washing of the resin and removal of the Fmoc group by treatment with piperidine/DMF, the second amino acid (Fmoc-Cys(Acm)) is introduced to continue sequence elongation. Fmoc protected amino acids are activated in situ using TBTU/HOBt and subsequently coupled to the resin over about 50 minutes. Diisopropylethylamine or collidine is used during coupling as an organic base. Completion of the coupling is indicated by ninhydrin test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps are repeated each time with another amino acid according to the peptide sequence. All amino acids used are Fmoc-Nalpha protected, except the last amino acid, Boc-D-Phe. Trifunctional amino acids are side chain protected as follows: Cys(Trt) and Cys(Acm), Tyr(tBu), and Lys(Boc). Three equivalents of the activated amino acids are used in the coupling reactions. At the end of the synthesis, the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS procedure starting from 2-Cl-Trt-chloride resin. The first amino acid (Fmoc-Thr(tBu)) is loaded on the resin as described in previous examples to obtain a loading of about 0.7 mmol/g of amino acid/resin. After washing of the resin and removal of the Fmoc group by treatment with piperidine/DMF, the second amino acid (Fmoc-Cys(Acm)) is introduced to continue sequence elongation. Fmoc protected amino acids are activated in situ using TBTU/HOBt and subsequently coupled to the resin over about 50 minutes. Diisopropylethylamine or collidine is used during coupling as an organic base. Completion of the coupling is indicated by ninhydrin test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps are repeated each time with another amino acid according to the peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Cys(Trt) and Cys(Acm), Tyr(tBu), Thr(tBu), and Lys(Boc). Three equivalents of the activated amino acids are used in the coupling reactions. At the end of the synthesis, the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Chelmical synthesis: Peptides were synthesized on a Rink amide resin, 0.45 mmol/g [Fmoc-Cys(Trityl)-Wang; Novabiochem, San Diego, Calif.] usinig N-(9-fluorenyl)methoxycarboxyl chemistry and standard side chain protection except on cysteine residues. Cysteine residues were protected in pairs with either S-trityl on the first and third cysteines or S-acetamidomethyl on the second and fourth cysteines. Amino acid derivatives were from Advanced Chemtech (Louisville, Ky.). The peptides were removed from the resin and precipitated, and a two-step oxidation protocol was used to selectively fold the peptides as described previously (Luo et al., 1999). Briefly, the first disulfide bridge was closed by dripping the peptide into an equal volume of 20 mM potassium feliicyanide and 0.1 M Tris, pH 7.5. The solution was allowed to react for 30 min, and the monocyclic peptide was purified by reverse-phase HPLC. Simultaneous removal of the S-acetamidomethyl groups and closure of the second disulfide bridge was carried out by iodine oxidation. The monocyclic peptide and HPLC eluent was dripped into an equal volume of iodine (10 mM) in H20/trifluoroacetic acid/acetonitrile (78:2:20 by volume) and allowed to react for 10 min. The reaction was terminated by the addition of ascorbic acid diluted 20-fold with 0.1percent trifluoroacetic acid and the bicyclic product purified by HPLC. Mass Spectrometry: Measurements were performed at the Salk Institute for Biological Studies (San Diego, Calif.) under the direction of Jean Rivier. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and liquid secondary ionization mass spectrometry were used. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 2h; | All the peptides were assembled manually on Rink amideMBHA resin (0.15g, 0.4 mmol/g) using the conventional Fmoc chemistry in a glass reaction vessel. The coupling reactions were carried out by adding a pre-activated solution of N-alpha-Fmoc-protected amino acid (3 equiv), HOBT (3 equiv), HBTU (3 equiv), and DIEA (6 equiv) in anhydrous DMF (10 mL/g resin) into the resin (1 equiv) and swirling for 2 h. The progress of the coupling was monitored by Kaiser test. The Fmoc protecting groups were removed by two 10 min treatments with a piperidine/DMF solution (20percent). The resin was washed by swirling in methanol (1 min, 2X), followed by DMF (1 min, 6X). The resin bound di-Cys(Acm) peptide was swirled with TI(F3CCOO)3 (2 equiv) in DMF to form the disulfide peptide, washed with DMF (1 min, 2X), and finally EPO <DP n="40"/>deblocked with 20percent piperidine in DMF. All the crude products were purified by semi-preparative HPLC and identified by both ES-MS and analytical HPLC. They showed the corresponding [MH]+, [MH2]2+ and even triple [MH3]3* peaks in their ES-MS spectra. A mixture of Cypate (211.5 mg, 0.3 mmol), DIC (38.0 mg,0.3 mmol), HOBT (41.0 mg, 0.3 mmol) and the resin bound disulfide RGD peptide (-0.06 mmol) in anhydrous DMF (3mL) was swirled for 8 h. The resin was filtered, washed with DMF (1 min, 3X), methanol (1 min, 2X), and dried under vacuum. The resin was cleaved with TFA (3 mL, 1.5 h, 5X). The TFA filtrate was concentrated and added to cold MBTE (10 mL). The solid precipitate was collected by filtration, washed with petroleum ether, and purified by semi-preparative HPLC to afford the desired products. The protected RGD peptide sequence, i.e. Fmoc-Cys(Acm)-Arg(Pbf)-Gly-Asp(OBut)-XAA-Cys(Acm) [SEQ ID NO:16], was first assembled on Rink amide MBHA resin (1 equiv) using the conventional Fmoc chemistry (Scheme 1). It was swirled with TI(F3CCOOH)3 (2 equiv) in DMF for 2 h to form the cyclic disulfide peptide, i.e., Fmoc-cyclo[Cys-Arg(Pbf)-Gly- Asp(OBut)-XAA-Cys] [SEQ ID NO:17] The conjugation of cypate with peptide usually afforded the monomeric and dimeric conjugates simultaneously. To increase the yield of monomeric product, we used 5 equivalent Cypate with 3.5 equivalent DIC and HOBT. Finally, TFA cleavage afforded the desired product, i.e., Cypate-cyclo(Cys-Arg-Gly-Asp-XAA-Cys)-NH2 [SEQ ID NO: 18]. The other cyclic disulfide RGD peptide analogs (Table 2) were synthesized using the same protocol. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide was carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from 2-Cl-Trt resin (50 g). The first amino acid (Fmoc-Cys(Acm)) was loaded onto the resin in a preliminary step to provide loading of about 0.7 mmol/g. After resin washing, a second amino acid (Fmoc-Pro) was introduced to start the first coupling step. Fmoc protected amino acid was activated in situ using TBTU/HOBt and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or Collidine were used during coupling as an organic base. Completion of the coupling was indicated by ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine was removed with 20percent piperidine in DMF for 20 min. These steps were repeated each time with another amino acid according to peptide sequence. All amino acids used were Fmoc-Nalpha protected except the last building block in the sequence, Trt-Mpa. Trifunctional amino acids were side chain protected as follows: Asp(tBu), Har(Pbf), and Cys(Acm). Three equivalents of the activated amino acids were employed in the coupling reactions. At the end of the synthesis the peptide-resin was washed with DMF, followed by DCM, and dried under vacuum to obtain 80 g dry peptide-resin. The peptide, prepared as described above, was cleaved from the resin by washing with a solution of 1percent TFA in DCM. The resulted solution was neutralized by addition of DIPEA and concentrated to about 10percent peptide content. Amidation of the C-terminus was achieved by activation of the carboxy terminus with DCC/HOBt and coupling with ammonia solution in IPA. After removal of the solvent the protected peptide was precipitated in ether and dried. The protecting groups were removed using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product was precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum to obtain 30 g product |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from 2-Cl-Trt resin. The first amino acid (Fmoc-Cys(Acm)) is loaded on the resin in a preliminary step to provide loading of about 0.7 mmol/g. After washing of the resin the Fmoc group is removed by treatment with piperidine/DMF solution and second amino acid (Fmoc-Ser(tBu)) is introduced to start the first coupling step. Fmoc protected amino acids are activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine are used during coupling as an organic base. Completion of the coupling is indicated by ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps were repeated each time with another amino acid according to peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Lys(Boc), Thr(tBu), Ser(tBu), Cys(Trt) and Cys(Acm). Three equivalents of the activated amino acids are employed in the coupling reactions. At the end of the synthesis the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. The peptide, prepared as described above, is cleaved from the resin accompanied with simultaneous deprotection of acid-labile protecting groups using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product is precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from 2-Cl-Trt resin. The first amino acid (Fmoc-Phe) is loaded on the resin in a preliminary step to provide loading of about 0.7 mmol/g. After washing of the resin the Fmoc group is removed by treatment with piperidine/DMF solution and second amino acid (Fmoc-Gly) is introduced to start the first coupling step. Fmoc protected amino acids are activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine are used during coupling as an organic base. Completion of the coupling is indicated by ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps were repeated each time with another amino acid according to peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Tyr(tBu), Arg(Pbf), Ser(tBu), Cys(Trt) and Cys(Acm). Three equivalents of the activated amino acids are employed in the coupling reactions. At the end of the synthesis the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. The peptide, prepared as described above, is cleaved from the resin accompanied with simultaneous deprotection of acid-labile protecting groups using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product is precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from Rink amide resin. After removal of the Fmoc protecting group from the resin the first amino acid (Fmoc-Thr(tBu)) is loaded on the resin in a regular coupling step to provide loading of about 0.7 mmol/g. After washing of the resin and removal of the Fmoc protecting group the second amino acid (Fmoc-Cys(Acm)) is introduced to start the second coupling step. Fmoc protected amino acids are activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine are used during coupling as an organic base. Completion of the coupling is indicated by ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps are repeated each time with another amino acid according to peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Tyr(tBu), Lys(Boc), Thr(tBu), Cys(Acm), and Cys(Trt). Three equivalents of the activated amino acids are used in the coupling reactions. At the end of the synthesis the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. The peptide, prepared as described above, is cleaved from the resin together with removal of acid-labile protecting groups using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product is precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum to obtain crude product. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from Rink amide resin (50 g). After removal of the Fmoc protecting group from the resin the first amino acid (Fmoc-Gly) is loaded on the resin in a regular coupling step to provide loading of about 0.7 mmol/g. After washing of the resin and removal of the Fmoc protecting group the second amino acid (Fmoc-Arg(Pbf)) is introduced to start the second coupling step. Fmoc protected amino acids are activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine are used during coupling as an organic base. Completion of the coupling is indicated by ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps are repeated each time with another amino acid according to peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Tyr(tBu), Arg(Pbf), Cys(Acm) and Cys(Trt). Asn and Gln are used unprotected on the amide group. Three equivalents of the activated amino acids are employed in the coupling reactions. At the end of the synthesis the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain 110 g dry peptide-resin. The peptide, prepared as described above, is cleaved from the resin together with removal of acid-labile protecting groups using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product is precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum to obtain 36 g product. Residual TFA <0.25percent. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide is carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from Rink amide resin. After removal of the Fmoc protecting group from the resin the first amino acid (Fmoc-Trp) is loaded on the resin in a regular coupling step to provide loading of about 0.7 mmol/g. After washing of the resin and removal of the Fmoc protecting group the second amino acid (Fmoc-Cys(Acm)) is introduced to start the second coupling step. Fmoc protected amino acids are activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine are used during coupling as an organic base. Completion of the coupling is indicated by Ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine is removed with 20percent piperidine in DMF for 20 min. These steps are repeated each time with another amino acid according to peptide sequence. All amino acids used are Fmoc-Nalpha protected. Trifunctional amino acids are side chain protected as follows: Lys(Boc), Tyr(tBu), Cys(Acm) and Cys(Trt). Three equivalents of the activated amino acids are used in the coupling reactions. At the end of the synthesis the peptide-resin is washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. The peptide, prepared as described above, is cleaved from the resin together with removal of acid-labile protecting groups using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product is precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum to obtain crude product. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide was carried out by a stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from Thr(t-Bu)-ol-2-Cl-Trt resin (250 g, loading of 0.7 mmol on 1 g of preloaded resin). After washing of the resin the second amino acid (Fmoc-Cys(Acm)) was introduced to start the first coupling step. Fmoc protected amino acid was activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine was used during coupling as an organic base. Completion of the coupling was indicated by Ninhydrine test. After washing of the resin, the Fmoc protecting group on the -amine was removed with 20percent piperidine in DMF for 20 min. These steps were repeated each time with another amino acid according to peptide sequence. All amino acids used were Fmoc-N protected except the last amino acid in the sequence, Boc-D-Phe. Trifunctional amino acids were side chain protected as follows: Thr(t-Bu), Cys(Trt), Cys(Acm), and Lys(Boc). Three equivalents of the activated amino acids were employed in the coupling reactions. At the end of the synthesis the peptide-resin was washed with DMF, followed by DCM, and dried under vacuum to obtain 510 g dry peptide-resin. The peptide, prepared as described above, was cleaved from the resin using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product was precipitated by the addition of 10 volumes of ether (MTBE), filtered and dried in vacuum to obtain 201.7 g powder. It was identified by LC/MS as H-D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys(Acm)-Thr-ol. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide was carried out by a regular stepwise Fmoc SPPS procedure starting from Rink amide resin (200 g). The first amino acid (Fmoc-Gly) was loaded on the resin by a regular coupling procedure after removal of the Fmoc group from the resin. After washing of the resin the second amino acid (Fmoc-D-Arg(Pbf)) was introduced to continue sequence elongation. Fmoc protected amino acids were activated in situ using TBTU/HOBt and subsequently coupled to the resin over about 50 minutes. Diisopropylethylamine or collidine were used during coupling as an organic base. Completion of the coupling was indicated by ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine was removed with 20percent piperidine in DMF for 20 min. These steps were repeated each time with another amino acid according to peptide sequence. All amino acids used were Fmoc-Nalpha protected except the last building block in the sequence, Trt-Mpa. Trifunctional amino acids were side chain protected as follows: Gln(Trt), D-Arg(Pbf), Tyr(tBu) and Cys(Acm). Three equivalents of the activated amino acids were employed in the coupling reactions. At the end of the synthesis the peptide-resin was washed with DMF, followed by DCM, and dried under vacuum to obtain 460 g dry peptide-resin. The peptide, prepared as described above, was cleaved from the resin using a 89percent TFA, 5.0percent Phenol, 1.0percent TIS, 2.5percent EDT, 2.5percent water solution for 1.5 hours at room temperature. The product was precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum to obtain 115.0 g powder. It was identified by LC/MS as Mpa-Tyr-Phe-Gln-Asn-Cys(Acm)-Pro-D-Arg-Gly-NH2 |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 20℃; for 1h; | Synthesis of the peptide is carried out by a regular stepwise "solution synthesis" method. The second amino acid (Fmoc-D-Arg(Pbf)-OH) is dissolved in DMF and pre-activated by addition of TBTU/HOBt in the presence of DIPEA. The first amino acid (Gly-NH2) is dissolved in DMF, is added, and the reaction continues for about 1 h at room temperature. DMF is removed under low pressure and the residue is dissolved in ethylacetate. The organic solution is washed several times with aqueous HCl (1N), water and, NaHCO3 (5percent). After the solution is dried over Na2SO4, the solvent is evaporated to obtain Fmoc-D-Arg(Pbf)-Gly-NH2. Fmoc group is removed by dissolution in piperidine/DMF (20percent). The solution is concentrated and the crude di-peptide is precipitated in cold ether. By a similar procedure the rest of amino acids are added sequentially to obtain final protected linear peptide. Fmoc protected amino acids are activated in situ using TBTU/HOBt and subsequently coupled to the growing peptide chain. Diisopropylethylamine or collidine are used during coupling as an organic base. Completion of the coupling is determined by HPLC or TLC test. These steps are repeated each time with another amino acid according to the peptide sequence. All amino acids used are Fmoc-Nalpha protected except the last building block in the sequence, Trt-Mpa. Trifunctional amino acids are side chain protected as follows: Gln(Trt), D-Arg(Pbf), Tyr(tBu) and Cys(Acm). The peptide, prepared as described above, is deprotected from its acid-labile protecting groups using a 91.5percent TFA, 1.0percent TIS, 2.5percent EDT, 5.0percent water solution for 1.5 hours at room temperature. The crude product, Mpa-Tyr-Phe-Gln-Asn-Cys(Acm)-Pro-D-Arg-Gly-NH2, is precipitated by the addition of 10 volumes of ether, filtered, and dried in a vacuum to obtain fine powder. The product is identified by LC/MS |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
> 99.5% | With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; for 0.833333h; | Synthesis of the peptide was carried out by a regular stepwise Fmoc SPPS (solid phase peptide synthesis) procedure starting from Rink amide resin. After removal of the Fmoc protecting group from the resin, the first amino acid (Fmoc-Gly) was loaded on the resin in a regular coupling step to provide loading of about 0.7 mmol/g. After washing the resin and removing the Fmoc protecting group, the second amino acid (Fmoc-Leu) was introduced to start the second coupling step. Fmoc protected amino acids were activated in situ using TBTU/HOBt (N-hydroxybenzotriazole) and subsequently coupled to the resin for 50 minutes. Diisopropylethylamine or collidine were used during coupling as an organic base. Completion of the coupling was indicated by Ninhydrine test. After washing of the resin, the Fmoc protecting group on the alpha-amine was removed with 20percent piperidine in DMF for 20 min. These steps were repeated each time with another amino acid according to peptide sequence. All amino acids used were Fmoc-Nalpha protected. Trifunctional amino acids were side chain protected as follows: Tyr(tBu), Cys(Acm) and Cys(Trt). Three equivalents of the activated amino acids were used in the coupling reactions. At the end of the synthesis the peptide-resin was washed with DMF, followed by DCM, and dried under vacuum to obtain dry peptide-resin. The peptide, prepared as described above, was cleaved from the resin together with removal of acid-labile protecting groups using a 95percent TFA, 2.5percent TIS, 2.5percent EDT solution for 2 hours at room temperature. The product was precipitated by the addition of 10 volumes of ether, filtered and dried in vacuum to obtain crude product. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Ca. 9.5 mg | 200 mg (~0.05 mmol) of the resin obtained above was mixed with 4 mL of 20percent piperidine in DMF (20percent) and swirled for 20 min as described above. The resulting resin was washed with methanol (2X) and DMF (3X). Fmoc-Cys(Trt) (3 equiv, 0.15mmol), HOBT (3 equiv, 0.15mmol), HBTU (3 equiv, 0.15mmol) and DIEA (6 equiv, 0.3mmol) in DMF (2mL) was added into the resin. The resulting mixture was swirled for 2 h, filtered, and washed with methanol (2X), DMF (2X), and methanol (3X). The resin obtained above was mixed with 4 mL of 20percent piperidine in DMF (20percent) and swirled for 20 min as described above. The resulting resin was washed with methanol (2X) and DMF (3X). Acetic anhydride (200 uL, 2.1 mmol), DIEA (200uL, 1.2 mmoL) in DMF (2 mL) was added and the resulting mixture was swirled for 1 h, filtered, and washed with methanol (2X), DMF (2X), and methanol (3X). Finally, the obtained resin was mixed with 4 mL of a TFA cleavage solution consisting of TFA/thioanisole/TIS/water (85:5:5:5) and swirled for 2 h, filtered, and washed with TFA. The filtrate was collected and combined and added into 15mL of cooled ether solution. Half of the precipitated solid was collected by centrifugation and purified by HPLC. The desired product (~9.5mg, ~30percent yield) was identified by analytical HPLC and ES-MS (observed m/z for [MH]+ and [MH2]2+: 1050.3638 and 525.7218). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Ca. 8 mg | 200 mg (~0.05 mmol) of the resin obtained above was mixed with 4 mL of 20percent piperidine in DMF (20percent) and swirled for 20 min as described above. The resulting resin was washed with methanol (2X) and DMF (3X). Fmoc-Cys(Trt) (3 equiv, 0.15mmol), HOBT (3 equiv, 0.15mmol), HBTU (3 equiv, 0.15mmol) and DIEA (6 equiv, 0.3mmol) in DMF (2mL) was added into the resin. The resulting mixture was swirled for 2 h, filtered, and washed with methanol (2X), DMF (2X), and methanol (3X). The resin obtained above was mixed with 4 mL of 20percent piperidine in DMF (20percent) and swirled for 20 min as described above. The resulting resin was washed with methanol (2X) and DMF (3X). A solution of DOTA(3OBut)-COOH (0.15 mmol), HOBT (0.15mmol), HBTU (0.15mmol), and DIEA (0.3 mmoL) in DMF (2 mL) was added and the resulting mixture was swirled for 2 h, filtered, and washed with methanol (2X), DMF (2X), and methanol (3X). Finally, the obtained resin was mixed with 4 mL of a TFA cleavage solution consisting of TFA/thioanisole/TIS/water (85:5:5:5) and swirled for 2 h, filtered, and washed with TFA. The filtrate was collected and combined and added into 15mL of cooled ether solution. After the precipitated solid was collected by centrifugation, half of the amount was purified by HPLC. The desired product (~8mg, ~15percent yield) was identified by analytical HPLC and ES-MS (observed m/z for [MH]+ and [MH2]2+: 1435.3887 and 716.6744). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
100% | With dmap; diisopropyl-carbodiimide; In dichloromethane; at 20℃; | The hydrophobic tag 2 (756.7 mg, 1.00 mmol) was dissolved in CH2Cl2 (20 mL). N-Fmoc-Cys(Acm)-OH (621.7 mg, 1.50 mmol), DMAP (1.2 mg, 0.0100 mmol), and DIC (189.4 mg, 1.50 mmol) were then added to the solution. The reaction mixture was stirred at room temperature until the reaction completed. After the completion, CH3CN was added to the reaction mixture to give 3 quantitatively as a precipitate. |
100% | With dmap; diisopropyl-carbodiimide; In dichloromethane; at 20℃; for 0.333333h; | To a solution of 2,4-(didocosyloxy)benzyl alcohol (referredas ?hydrophobic tag? in the text) (1.51 g, 2.00 mmol) in dichloromethane (20mL), Fmoc-Cys(Acm)-OH (Acm: acetamidomethyl, 1.24 g, 3.00 mmol), N,N?-diisopropylcarbodiimide (DIPCI)(379 mg, 3.00 mmol), and 4-dimethylaminopyridine (DMAP) (12.2 mg, 0.10 mmol)were added. After stirring the resulting reaction mixture at room temperature for 20 min, large excess methanol was added, yielding a white precipitate. The precipitate was collected and washed with acetonitrile repeatedly to give the desired product quantitatively. Further peptide elongation cycle consists of Fmoc deprotection step using 2percent 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 2percentpiperidine in tetrahydrofuran (20 mL) (10 min at room temperature) and coupling step using a protected amino acid (1.20 mol equiv.),N-[1-(cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino(morpholino)]uronium hexafluorophosphate (COMU) (1.20 mol equiv.), and N,N-diisopropylethylamine(DIPEA) (2.40 mol equiv.) in tetrahydrofuran (20 mL) (20 min at room temperature). Some chemical structures of above mentioned substrates are shown in Scheme S1. Each tagged tripeptides (1, 3, 4) were characterized by HPLC and HRMS. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine; In tetrahydrofuran; at 20℃; | General procedure: H-Peptide-O-TAG was dissolved in THF (20 mL). N-Fmoc- or -Boc AAs (1.2 mol equiv.), HBTU (1.2 mol equiv.), HOBt (1.2 mol equiv.), and DIPEA (2.4 mol equiv.) were then added to the solution. The reaction mixture was stirred at room temperature until the reaction completed. After the completion, CH3CN was added to the reaction mixture to give N-Fmoc or -Boc-AA-Peptide-O-TAG quantitatively as a precipitate. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine; In tetrahydrofuran; at 20℃; | General procedure: The tagged peptide was dissolved in THF (20 mL). Amino acid (1.2 mol equiv), HBTU (1.2 mol equiv), HOBt (1.2 mol equiv), and DIPEA (2.4 mol equiv) were then added to the solution. The reaction mixture was stirred at room temperature until the reaction completed. After the completion, MeCN was added to the reaction mixture to give the coupled product as a precipitate. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
A solution of 2.58 grams of cesium carbonate in 2.5 ml of water was slowly added to a solution of 3.48 grams of Boc-L-Threonine (Bachem Calif., Torrance, Calif.) dissolved in 7 ml of methanol. The resulting mixture was stirred for about 1 hour at room temperature, and then all methanol and water were removed under reduced pressure yielding a dry powder of cesium salt of Boc-L-Threonine. 10 grams of Merrifield resin (chloromethylated polystyrene; 200-400 mesh, chloride incorporation of 1.3 meq/gram, Advanced ChemTech, Louisville, Ky.) was rinsed with dichloromethane (DCM), methanol (MeOH), and dimethylformamide (DMF) (2×70 ml each). The Boc-L-Threonine cesium salt powder was dissolved in 60 ml of dry DMF, and the solution was combined with the above washed resin. The slurry was gently mixed at about 50°-60° C. for about 85 to 90 hours under nitrogen atmosphere. The resin was filtered and rinsed well with DMF, deionized water, and finally MeOH. The Boc-Threonine resin was dried under reduced pressure at about 40° C. (Threonine incorporation=0.85±0.15 meq/gram of dry resin). B) H-D-beta-Nal-Cys(Acm)-Tyr-D-Trp-Lys-Val-Cys(Acm)-Thr-Resin (0085) 2.0 grams of Boc-Threonine resin from step A was added to a 50 ml glass reactor with a coarse sintered glass filter bottom (batch scale=1.74 mmole). The resin was rinsed two times for about 5 minutes each with DCM (20 ml), deblocked with 25percent TFA/DCM (30 ml) once for about 2 minutes and once for about 25 minutes, rinsed three times for about 2 minutes with DCM (20 ml), isopropanol (IPA) (20 ml), and DCM (20 ml), neutralized two times for about 5 minutes with 10percent triethylamine/DCM (20 ml), rinsed three times for about 2 minutes with DCM, and rinsed once for about 5 minutes with DMF (20 ml). (0086) The deblocked resin was coupled with 1.8 grams (4.35 mmole, 2.5 eq.) of Fmoc-L-Cysteine(Acm) (Bachem, Calif.) and 683 mul (4.35 mmole, 2.5 eq) diisopropylcarbodiimide (DIC) in 14 ml of 2:1 DCM/DMF for about 1 hour. The coupled resin was rinsed once for about 3 minutes with DMF (20 ml), three times for about 2 minutes with isopropanol (IPA), and three times for about 2 minutes with DCM (20 ml). The coupling was checked by Kaiser ninhydrin method. (0087) The coupled resin was then rinsed once with DMF and then deblocked with a solution of piperidine in DMF. The deblocked coupled resin was then rinsed with DMF and several times with both MeOH and DCM. The coupled resin was rinsed once for about 3 minutes with DMF (20 ml), three times for about 2 minutes with isopropanol (IPA) (20 ml), and three times for about 2 minutes each with DCM (20 ml). The coupling was checked by Kaiser ninhydrin method. (0088) Each of the following protected amino acids were coupled with the rinsed resin using DIC in DMF/DCM and deblocked in the following order as described above: Fmoc-L-Valine, Fmoc-L-Lysine(Boc), Fmoc-D-Tryptophan, Fmoc-L-Tyrosine(O-t-Bu), and Fmoc-L-Cysteine(Acm) (all from Bachem Calif.), Boc-D-2-Naphthylalanine (Synthetech, Albany, Oreg.). (0089) The completed peptide chain was deblocked and deprotected twice with 75:20:5 DCM/TFA/anisole (30 ml) for about 2 minutes and about 25 minutes, rinsed three times for about 2 minutes each with DCM (20 ml), IPA (10 ml), and DCM (20 ml), neutralized two times for about 5 minutes with 10percent triethylamine/DCM (20 ml), and rinsed three times for about 2 minutes with DCM (20 ml) and MeOH (20 ml). The resin was dried under reduced pressure. Dry weight=3.91 grams (103percent of theory). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
94% | In general, linear peptide 18 was synthesized by the solid-phase synthesis strategy employing N-(9-fluorenyl)methoxycarbonyl (Fmoc)-based chemistry and Fmoc-amino acid building blocks. 1-Hydroxy-benzotriazole (HOBt), N,N-diisopropylethylamine (DIPEA), diisoprropylcarbodiimide (DIC) were used as coupling and activating reagents, respectively. Synthesis (SPPS) carried out on the 2-chlortritilchlorid-resin.2-Chlorotritylchloride resin (0.56 g, 0.3 mmol) was swelled in DCM for about 15 min. The solvent excess was filtered off. The swelling and filtration steps were repeated for 2 more times before the coupling reactions. Fmoc-Thr(tert-Bu)-OH 12 (0.616 g, 1.55 mmol) was attached to Trt(2-Cl)-resin (1.0 g., 1.55 mmol) for 1.5 h in the presence of DIPEA (0.64 mL, 3.87 mmol) in methylene chloride (Scheme 2). After the coupling was completed, the reaction solution was filtered off, and the resin was washed twice with a mixture DCM/MeOH/DIPEA (17:2:1 v/v/v). Then the resin was washed with DMF (2 × 20 mL). After Fmoc deprotection with 25% diethylamine in DMF, the resin was washed with DMF (5 × 15 mL). Amino acids Fmoc-Cys(Acm)-OH (1.29 g, 3.1 mmol), Fmoc-Thr(tert-Bu)-OH (1.23 g, 3.1 mmol), compound 11 (1.31 g, 3.1 mmol), Fmoc-D-Trp-OH (1.32 g, 3.1 mmol), Fmoc-Phe-OH (1.20 g, 3.1 mmol), Fmoc-Cys(Acm)-OH (1.28 g, 3.1 mmol), Fmoc-D-Phe (0.82 g, 3.1 mmol) were coupled in the presence of DIC (0.53 ml, 3.41 mmol) and HOBt (0.46 g, 3.41 mmol) as condensing agents in DMF. Coupling time was 2 h. After the coupling was completed, the reaction mixture was filtered off, and the resin was washed with DMF (5 × 20 mL). Peptide cleavage from the resin was performed using 1% TFA in DCM |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
All peptides were synthesized manually by Fmoc solid-phase methodology using DIC and HOAt as coupling reagents. In brief, resin (294 mg, 0.34 mmol/g) was placed into a 10 mL HSW polypropylene syringe with frits on the bottom (Torviq, Niles, Mich.) and swollen in DCM (5 mL) for 30 min. After removal of the Fmoc protecting group (25% piperidine in NMP, 5 mL, 5 and 10 min), the resin was washed four times with NMP (5 mL per wash) and DCM (5 mL per wash), and the individual amino acids were coupled to the resin. For each coupling, 3 equivalents (0.3 mmol) of the amino acid, HOAt, and DIC were used, with 10 min preactivation in NMP. All couplings were performed for 1 h and monitored to completion by either the Kaiser test or the chloranil test. If necessary, the N-terminal amino group was acetylated with 20 equivalents of acetic anhydride and 2 equivalents of DIPEA in 5 mL of DCM for 30 min. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Starting from Fmoc-Rink Amide MBHA resin (0.39 mmol/g, 128 mg, 50 mumol), Fmoc-Gly-(Et)Cys(Trt)-Arg(Pbf)-Arg(Pbf)-NH-resin was prepared by the same manner as described above. The peptide chain corresponding to the RTD-1 sequence was elongated manually by DCC-HOBt method, and H-Phe-Cys(But)-Arg(Pbf)-Cys(Acm)-Leu-Cys(Tfe-Pocam)- Arg(Pbf)-Arg(Pbf)-Gly-Val-Cys(Tfe-Pocam)-Arg(Pbf)-Cys(Acm)-Ile-Cys(But)-Thr(But)-Arg(Pbf)-Gly-(Et)Cys(Trt)-Arg(Pbf)-Arg(Pbf)-NH-resin (373 mg) was obtained. A part of the resin (53 mg) was treated with a TFA cocktail (TFA/thioanisole/H2O/phenol/triisopropylsilane, 82.5/5/5/5/2.5, 1 mL) at room temperature for 2 h. TFA was removed under nitrogen stream and the peptide was precipitated with diethyl ether. After washing twice with ether, the precipitant was dried under vacuum. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
19% | Azidoacetyl-JR11 was synthesized via the Nalpha-Fmoc solid-phase peptide synthesis strategy starting from D-Tyr-Rink Amide MBHA resin. The resin was treated with 20% piperidine in DMF to remove the Nalpha-Fmoc protecting group. The following Fmoc-protected amino acids (3 equivalents) including Fmoc-Cys(Acm)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-D-Aph(Cbm)-OH, Fmoc-Aph(Hor)-OH, <strong>[168300-88-7]Fmoc-D-Cys(Acm)-OH</strong>, Fmoc-Cl-Phe-OH were subsequently coupled to the sequence in correct order. The coupling was carried out in NMP with standard in situ activating reagent HBTU/HOBT (3 equivalents) in the presence of DIEA (6 equivalents). Cyclization was performed by incubation of the resin with 2 equivalents of thallium(III) trifluoroacetate in DMF at room temperature for 90 min. Finally, azidoacetic acid (10 equivalents) was pre-activated with DIC (5 equivalents) in DCM for 10 min, filtered, and then coupled to the peptide sequence to provide the azide functional group at the N-terminus for click reaction. The peptide was de-protected and simultaneously cleaved from the resin by the treatment of a cocktail of trifluoroacetic acid/water/triisopropylsilane (95:2.5:2.5). After filtration, the peptide was precipitated by the addition of cold diethyl ether to the TFA solution. The crude product was filtered, dried, and purified by HPLC (Luna C18 semi-prep column, 4.5 mL/min, 30-35% MeCN (0.1% TFA) in water (0.1% TFA), RT=10). Azidoacetyl-JR11 was obtained in 19% yield. ESI-MS: calculated for Azidoacetyl-JR11 C60H73ClN18O15S2 1384.5. found [M+H]+ 1385.8. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
15% | Azidoacetyl-LM3 was synthesized via the Nalpha-Fmoc solid-phase peptide synthesis strategy starting from D-Tyr-Rink Amide MBHA resin. The resin was treated with 20% piperidine in DMF to remove the Nalpha-Fmoc protecting group. The following Fmoc-protected amino acids (3 equivalents) including Fmoc-Cys(Acm)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-D-Phe(Cbm)-OH, Fmoc-Tyr(tBu)-OH, <strong>[168300-88-7]Fmoc-D-Cys(Acm)-OH</strong>, Fmoc-Cl-Phe-OH were subsequently coupled to the sequence in correct order. The coupling was carried out in NMP with standard in situ activating reagent HBTU/HOBT (3 equivalents) in the presence of DIEA (6 equivalents). Cyclization was performed by incubation of the resin with 2 equivalents of thallium(III) trifluoroacetate in DMF at room temperature for 90 min. Bromoacetic acid (40 equivalents) was pre-activated with DIC (20 equivalents) in DCM for 10 min, filtered, and then coupled to the peptide sequence. Finally, the resin was treated with sodium azide (27.5 equivalents) in DMSO to provide the azide functional group at the N-terminus for click reaction. The peptide was de-protected and simultaneously cleaved from the resin by the treatment of a cocktail of trifluoroacetic acid/water/triisopropylsilane (95:2.5:2.5). After filtration, the peptide was precipitated by the addition of cold diethyl ether to the TFA solution. The crude product was filtered, dried, and purified by HPLC (Luna C18 semi-prep column, 4.5 mL/min, 20-35% MeCN (0.1% TFA) in Water (0.1% TFA) in 30 min, RT=30). Azidoacetyl-LM3 was obtained in 15% yield. ESI-MS: calculated for Azidoacetyl-LM3 C55H68ClN15O13S2 1245.4. found [M+H]+ 1246.9. |
Tags: 86060-81-3 synthesis path| 86060-81-3 SDS| 86060-81-3 COA| 86060-81-3 purity| 86060-81-3 application| 86060-81-3 NMR| 86060-81-3 COA| 86060-81-3 structure
[ 159680-21-4 ]
(S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-(((2-phenylacetamido)methyl)thio)propanoic acid
Similarity: 0.93
Precautionary Statements-General | |
Code | Phrase |
P101 | If medical advice is needed,have product container or label at hand. |
P102 | Keep out of reach of children. |
P103 | Read label before use |
Prevention | |
Code | Phrase |
P201 | Obtain special instructions before use. |
P202 | Do not handle until all safety precautions have been read and understood. |
P210 | Keep away from heat/sparks/open flames/hot surfaces. - No smoking. |
P211 | Do not spray on an open flame or other ignition source. |
P220 | Keep/Store away from clothing/combustible materials. |
P221 | Take any precaution to avoid mixing with combustibles |
P222 | Do not allow contact with air. |
P223 | Keep away from any possible contact with water, because of violent reaction and possible flash fire. |
P230 | Keep wetted |
P231 | Handle under inert gas. |
P232 | Protect from moisture. |
P233 | Keep container tightly closed. |
P234 | Keep only in original container. |
P235 | Keep cool |
P240 | Ground/bond container and receiving equipment. |
P241 | Use explosion-proof electrical/ventilating/lighting/equipment. |
P242 | Use only non-sparking tools. |
P243 | Take precautionary measures against static discharge. |
P244 | Keep reduction valves free from grease and oil. |
P250 | Do not subject to grinding/shock/friction. |
P251 | Pressurized container: Do not pierce or burn, even after use. |
P260 | Do not breathe dust/fume/gas/mist/vapours/spray. |
P261 | Avoid breathing dust/fume/gas/mist/vapours/spray. |
P262 | Do not get in eyes, on skin, or on clothing. |
P263 | Avoid contact during pregnancy/while nursing. |
P264 | Wash hands thoroughly after handling. |
P265 | Wash skin thouroughly after handling. |
P270 | Do not eat, drink or smoke when using this product. |
P271 | Use only outdoors or in a well-ventilated area. |
P272 | Contaminated work clothing should not be allowed out of the workplace. |
P273 | Avoid release to the environment. |
P280 | Wear protective gloves/protective clothing/eye protection/face protection. |
P281 | Use personal protective equipment as required. |
P282 | Wear cold insulating gloves/face shield/eye protection. |
P283 | Wear fire/flame resistant/retardant clothing. |
P284 | Wear respiratory protection. |
P285 | In case of inadequate ventilation wear respiratory protection. |
P231 + P232 | Handle under inert gas. Protect from moisture. |
P235 + P410 | Keep cool. Protect from sunlight. |
Response | |
Code | Phrase |
P301 | IF SWALLOWED: |
P304 | IF INHALED: |
P305 | IF IN EYES: |
P306 | IF ON CLOTHING: |
P307 | IF exposed: |
P308 | IF exposed or concerned: |
P309 | IF exposed or if you feel unwell: |
P310 | Immediately call a POISON CENTER or doctor/physician. |
P311 | Call a POISON CENTER or doctor/physician. |
P312 | Call a POISON CENTER or doctor/physician if you feel unwell. |
P313 | Get medical advice/attention. |
P314 | Get medical advice/attention if you feel unwell. |
P315 | Get immediate medical advice/attention. |
P320 | |
P302 + P352 | IF ON SKIN: wash with plenty of soap and water. |
P321 | |
P322 | |
P330 | Rinse mouth. |
P331 | Do NOT induce vomiting. |
P332 | IF SKIN irritation occurs: |
P333 | If skin irritation or rash occurs: |
P334 | Immerse in cool water/wrap n wet bandages. |
P335 | Brush off loose particles from skin. |
P336 | Thaw frosted parts with lukewarm water. Do not rub affected area. |
P337 | If eye irritation persists: |
P338 | Remove contact lenses, if present and easy to do. Continue rinsing. |
P340 | Remove victim to fresh air and keep at rest in a position comfortable for breathing. |
P341 | If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable for breathing. |
P342 | If experiencing respiratory symptoms: |
P350 | Gently wash with plenty of soap and water. |
P351 | Rinse cautiously with water for several minutes. |
P352 | Wash with plenty of soap and water. |
P353 | Rinse skin with water/shower. |
P360 | Rinse immediately contaminated clothing and skin with plenty of water before removing clothes. |
P361 | Remove/Take off immediately all contaminated clothing. |
P362 | Take off contaminated clothing and wash before reuse. |
P363 | Wash contaminated clothing before reuse. |
P370 | In case of fire: |
P371 | In case of major fire and large quantities: |
P372 | Explosion risk in case of fire. |
P373 | DO NOT fight fire when fire reaches explosives. |
P374 | Fight fire with normal precautions from a reasonable distance. |
P376 | Stop leak if safe to do so. Oxidising gases (section 2.4) 1 |
P377 | Leaking gas fire: Do not extinguish, unless leak can be stopped safely. |
P378 | |
P380 | Evacuate area. |
P381 | Eliminate all ignition sources if safe to do so. |
P390 | Absorb spillage to prevent material damage. |
P391 | Collect spillage. Hazardous to the aquatic environment |
P301 + P310 | IF SWALLOWED: Immediately call a POISON CENTER or doctor/physician. |
P301 + P312 | IF SWALLOWED: call a POISON CENTER or doctor/physician IF you feel unwell. |
P301 + P330 + P331 | IF SWALLOWED: Rinse mouth. Do NOT induce vomiting. |
P302 + P334 | IF ON SKIN: Immerse in cool water/wrap in wet bandages. |
P302 + P350 | IF ON SKIN: Gently wash with plenty of soap and water. |
P303 + P361 + P353 | IF ON SKIN (or hair): Remove/Take off Immediately all contaminated clothing. Rinse SKIN with water/shower. |
P304 + P312 | IF INHALED: Call a POISON CENTER or doctor/physician if you feel unwell. |
P304 + P340 | IF INHALED: Remove victim to fresh air and Keep at rest in a position comfortable for breathing. |
P304 + P341 | IF INHALED: If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable for breathing. |
P305 + P351 + P338 | IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. |
P306 + P360 | IF ON CLOTHING: Rinse Immediately contaminated CLOTHING and SKIN with plenty of water before removing clothes. |
P307 + P311 | IF exposed: call a POISON CENTER or doctor/physician. |
P308 + P313 | IF exposed or concerned: Get medical advice/attention. |
P309 + P311 | IF exposed or if you feel unwell: call a POISON CENTER or doctor/physician. |
P332 + P313 | IF SKIN irritation occurs: Get medical advice/attention. |
P333 + P313 | IF SKIN irritation or rash occurs: Get medical advice/attention. |
P335 + P334 | Brush off loose particles from skin. Immerse in cool water/wrap in wet bandages. |
P337 + P313 | IF eye irritation persists: Get medical advice/attention. |
P342 + P311 | IF experiencing respiratory symptoms: call a POISON CENTER or doctor/physician. |
P370 + P376 | In case of fire: Stop leak if safe to Do so. |
P370 + P378 | In case of fire: |
P370 + P380 | In case of fire: Evacuate area. |
P370 + P380 + P375 | In case of fire: Evacuate area. Fight fire remotely due to the risk of explosion. |
P371 + P380 + P375 | In case of major fire and large quantities: Evacuate area. Fight fire remotely due to the risk of explosion. |
Storage | |
Code | Phrase |
P401 | |
P402 | Store in a dry place. |
P403 | Store in a well-ventilated place. |
P404 | Store in a closed container. |
P405 | Store locked up. |
P406 | Store in corrosive resistant/ container with a resistant inner liner. |
P407 | Maintain air gap between stacks/pallets. |
P410 | Protect from sunlight. |
P411 | |
P412 | Do not expose to temperatures exceeding 50 oC/ 122 oF. |
P413 | |
P420 | Store away from other materials. |
P422 | |
P402 + P404 | Store in a dry place. Store in a closed container. |
P403 + P233 | Store in a well-ventilated place. Keep container tightly closed. |
P403 + P235 | Store in a well-ventilated place. Keep cool. |
P410 + P403 | Protect from sunlight. Store in a well-ventilated place. |
P410 + P412 | Protect from sunlight. Do not expose to temperatures exceeding 50 oC/122oF. |
P411 + P235 | Keep cool. |
Disposal | |
Code | Phrase |
P501 | Dispose of contents/container to ... |
P502 | Refer to manufacturer/supplier for information on recovery/recycling |
Physical hazards | |
Code | Phrase |
H200 | Unstable explosive |
H201 | Explosive; mass explosion hazard |
H202 | Explosive; severe projection hazard |
H203 | Explosive; fire, blast or projection hazard |
H204 | Fire or projection hazard |
H205 | May mass explode in fire |
H220 | Extremely flammable gas |
H221 | Flammable gas |
H222 | Extremely flammable aerosol |
H223 | Flammable aerosol |
H224 | Extremely flammable liquid and vapour |
H225 | Highly flammable liquid and vapour |
H226 | Flammable liquid and vapour |
H227 | Combustible liquid |
H228 | Flammable solid |
H229 | Pressurized container: may burst if heated |
H230 | May react explosively even in the absence of air |
H231 | May react explosively even in the absence of air at elevated pressure and/or temperature |
H240 | Heating may cause an explosion |
H241 | Heating may cause a fire or explosion |
H242 | Heating may cause a fire |
H250 | Catches fire spontaneously if exposed to air |
H251 | Self-heating; may catch fire |
H252 | Self-heating in large quantities; may catch fire |
H260 | In contact with water releases flammable gases which may ignite spontaneously |
H261 | In contact with water releases flammable gas |
H270 | May cause or intensify fire; oxidizer |
H271 | May cause fire or explosion; strong oxidizer |
H272 | May intensify fire; oxidizer |
H280 | Contains gas under pressure; may explode if heated |
H281 | Contains refrigerated gas; may cause cryogenic burns or injury |
H290 | May be corrosive to metals |
Health hazards | |
Code | Phrase |
H300 | Fatal if swallowed |
H301 | Toxic if swallowed |
H302 | Harmful if swallowed |
H303 | May be harmful if swallowed |
H304 | May be fatal if swallowed and enters airways |
H305 | May be harmful if swallowed and enters airways |
H310 | Fatal in contact with skin |
H311 | Toxic in contact with skin |
H312 | Harmful in contact with skin |
H313 | May be harmful in contact with skin |
H314 | Causes severe skin burns and eye damage |
H315 | Causes skin irritation |
H316 | Causes mild skin irritation |
H317 | May cause an allergic skin reaction |
H318 | Causes serious eye damage |
H319 | Causes serious eye irritation |
H320 | Causes eye irritation |
H330 | Fatal if inhaled |
H331 | Toxic if inhaled |
H332 | Harmful if inhaled |
H333 | May be harmful if inhaled |
H334 | May cause allergy or asthma symptoms or breathing difficulties if inhaled |
H335 | May cause respiratory irritation |
H336 | May cause drowsiness or dizziness |
H340 | May cause genetic defects |
H341 | Suspected of causing genetic defects |
H350 | May cause cancer |
H351 | Suspected of causing cancer |
H360 | May damage fertility or the unborn child |
H361 | Suspected of damaging fertility or the unborn child |
H361d | Suspected of damaging the unborn child |
H362 | May cause harm to breast-fed children |
H370 | Causes damage to organs |
H371 | May cause damage to organs |
H372 | Causes damage to organs through prolonged or repeated exposure |
H373 | May cause damage to organs through prolonged or repeated exposure |
Environmental hazards | |
Code | Phrase |
H400 | Very toxic to aquatic life |
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.
Home
* Country/Region
* Quantity Required :
* Cat. No.:
* CAS No :
* Product Name :
* Additional Information :