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NPC 15199 is an anti-inflammatory agent. It increases intracellular Ca2+ levels.
Synonyms: Fmoc-leucine; N-FMOC-leucine; FMOC-Leu
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*For Research Use Only !
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Agrawal, Anushka ; Euliano, Erin M ; Pogostin, Brett H , et al. Cel. Mol. Bioeng.,2024,17,441-451.
Abstract: Introduction Multidomain peptides (MDPs) are amino acid sequences that self-assemble to form supramolecular hydrogels under physiological conditions that have shown promise for a number of biomedical applications. K2(SL)6K2 (“K2”), a widely studied MDP, has demonstrated the ability to enhance the humoral immune response to co-delivered antigen. Herein, we sought to explore the in vitro and in vivo properties of a peptide with the same sequence but opposite chirality (D-K2) since peptides composed of D-amino acids are resistant to protease degradation and potentially more immunostimulatory than their canonical counterparts. Methods K2 and D-K2 hydrogels were characterized and evaluated in vitro using circular dichroism, rheology, cryo-electron microscopy, and fuorescence recovery after photobleaching studies. In vivo experiments in SKH-1 mice were conducted to evaluate both ovalbumin release from the hydrogels and hydrogel degradation. The injection site of the hydrogels was analyzed using histology and humoral immunity was assessed by ELISA. Results In vitro, the enantiomeric hydrogels exhibited similar rheological properties, and fuorescence recovery after pho_x005f_x0002_tobleaching experiments demonstrated that the difusion of ovalbumin (OVA), a model antigen, was similar within both hydrogels. In vivo, K2 and D-K2 peptide hydrogels had similar OVA release rates, both releasing 89% of the antigen within 8 days. Both hydrogels elicited a similar antigen-specifc humoral immune response. However, the in vivo degradation of the D-K2 hydrogel progressed signifcantly slower than K2. After 4 weeks in vivo, only 23±7% of the K2 hydrogel remained at the injection site compared to 94±7% of the D-K2 hydrogel, likely due to their diferent protease susceptibilities. Conclusion Taken together, these data suggest that peptide chirality can be a useful tool for increasing hydrogel residence time for biomedical applications that would beneft from long persistence times and that, if an antigen releases over a suf_x005f_x0002_fciently short period, release can be largely independent of degradation rate, though slower-difusing payloads may exhibit degradation rate dependence.
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Keywords: Hydrogel ; Chirality ; Degradation ; Drug delivery ; Peptides ; Adjuvants
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Purchased from AmBeed: 73724-45-5 ; 35661-60-0 ; 114360-54-2 ; 110990-08-4 ; 116861-26-8
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CAS No. : | 35661-60-0 |
Formula : | C21H23NO4 |
M.W : | 353.41 |
SMILES Code : | CC(C)C[C@@H](C(O)=O)NC(OCC1C2=C(C3=C1C=CC=C3)C=CC=C2)=O |
Synonyms : |
Fmoc-leucine; N-FMOC-leucine; FMOC-Leu
|
MDL No. : | MFCD00037133 |
InChI Key : | CBPJQFCAFFNICX-IBGZPJMESA-N |
Pubchem ID : | 1549133 |
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 |
---|---|---|
All peptides were synthesized on a 0.2 mmol scale using manual Fmoc-SPPS chemistry under flow using a 3 minute cycle for each amino acid. Specifically, all reagents and solvents are delivered to a stainless steel reactor containing resins at a constant flow rate using HPLC pump; temperature of the reactor was maintained at 60 °C during the synthesis using water bath. Procedure for each amino acid coupling cycle included a 30 second coupling with 1 mmol Fmoc-protected amino acid, 1 mmol HBTU, and 500 of diisopropyl ethyl amine (DIEA) in 2.5 mL of DMF at a flow rate of 6 mL/min (note that for coupling of cysteine and tryptophan, 190 of DIEA was used to prevent racemization); 1 minute wash with DMF at a flow rate of 20 mL/min; 20 second deprotection with 50percent (v/v) piperidine in DMF at a flow rate of 20 mL/min; and 1 minute wash with DMF at a flow rate was 20 mL/min. After completion of the stepwise SPPS, the resin was washed thoroughly with DCM and dried under vacuum. The peptide is simultaneously cleaved from the resin and side-chain deprotected by treatment with 2.5percent (v/v) water, 2.5percent (v/v) 1 ,2- ethanedithiol (EDT), and 1percent (v/v) triisoproprylsilane in neat trifluoroacetic acid (TFA) for 2 hours at room temperature. The resulting solution containing peptide was evaporated by blowing a stream of nitrogen gas over its surface for 15 minutes, then triturated and washed with cold diethyl ether three times. The obtained gummy-like solid was dissolved in 50percent H20: 50percent acetonitrile containing 0.1percent TFA and lyophilized. These same solvent compositions were used in majority of experiments and will be referred to as A: 0.1percent TFA in H20 and B: 0.1percent TFA in acetonitrile. c. Peptide Purification The crude peptide was dissolved in 95percent A: 5percent B with 6 M guanidinium hydrochloride and purified by semi-preparative RP-HPLC (Agilent Zorbax SB C18 column: 21.2 x 250 mm, 7 mutaueta, linear gradient: 5-50percent B over 90 min, flow rate: 5 mL/min). 1 of each HPLC fraction was mixed with 1 mu^ of alpha-cyano-4-hydroxycinnamic acid (CHCA) matrix in 75percent A: 25percent B, spotted with MALDI, and checked for fractions with desired molecular mass. The purity of fractions was confirmed by analytical RP-HPLC (Agilent Zorbax SB C3 column: 2.1 x 150 mm, 5 muiotaeta, gradient: 0-2 minutes 5percent B, 2-11 minutes 5- 65percent B, 11-12 minutes 65percent B, flow rate: 0.8 mL/min). HPLC fractions containing only product materials were confirmed by LC-MS analysis, combined, and then lyophilized. Peptides synthesized using fast flow-based SPPS and purified by RP-HPLC are listed in Table SI . |
Tags: 35661-60-0 synthesis path| 35661-60-0 SDS| 35661-60-0 COA| 35661-60-0 purity| 35661-60-0 application| 35661-60-0 NMR| 35661-60-0 COA| 35661-60-0 structure
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