Peptide Having Anti-Diabetic And Anti-Obesity Effects, And Use Thereof
An anti-diabetes and diabetes technology, applied in the direction of medical preparations containing active ingredients, obesity gene products, peptides, etc., can solve the problems of high treatment costs and difficult treatment of diabetes, and achieve the reduction of cholesterol values, excellent functions, and blood sugar reduction Effect
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Synthetic example 1
[0077] Synthesis Example 1: Peptide Synthesis
[0078] The chlorotrityl chloride resin (Chloro trityl chloride resin; CTL resin, Novabiochem Cat No.01-64-0021) of 700mg is put into reaction vessel, and the methylene chloride (MC, methylenechloride) that adds 10ml is stirred 3 minute. The solution was removed, and after 10 ml of dimethylformamide (DMF, DimethylFormamide) was put in to stir for 3 minutes, the solvent was removed again. Put 10ml of dichloromethane (DCM, Dichloromethane) solution into the reactor, and put 200mmol of fluorenylmethoxycarbonyl-Fmoc-Asn (Trt)-OH) (Bachem company (Bachem), Switzerland (Swiss)) and 400 mmole of N,N-diisopropylethylamine (DIEA, N,N-Diisopropylethylamine) was then stirred to dissolve them uniformly, and then stirred for 1 hour to carry out the reaction. Wash after the reaction, dissolve methanol and DIEA (2:1) in DCM for 10 minutes of reaction, and wash with excess DCM / DMF (1:1). The solution was removed, and after stirring for 3 minut...
Embodiment 1
[0082] Example 1: Evaluation of lipogenesis inhibitory activity
[0083] 1-1. Inhibition of lipid accumulation in preadipocytes (Oil red O staining)
[0084] After culturing 3T3-L1 cells as pre-adipocytes to a confluent state, they were replaced with 1-methyl-3-isobutyric acid containing 10 μg / ml insulin, 0.1 μM dexamethasone and 0.5 μM The differentiation medium containing xanthine (IBMX) and the peptides were treated according to the concentration and cultured for 2 days, and then the medium containing 10 μg / ml insulin was exchanged every 2 days. After 10 days of differentiation induction, in order to confirm the intracellular small Oil red O staining was carried out using droplets. The prepared 3T3-L1 preadipocytes were washed with phosphate buffered saline (PBS), fixed with 3.7% formalin for 1 hour, washed with 60% isopropanol, treated with Oil Red O solution and Stain for 20 minutes at room temperature. After staining, the Oil Red O solution was removed, washed three t...
Embodiment 2
[0096] Example 2: Evaluation of lipolytic activity
[0097] 2-1. Increased expression of genes involved in lipid breakdown
[0098] Press 3×10 5 Cell density of cells / well, 3T3-L1 (preadipocytes) were seeded in 6-well plates. After culturing for 24 hours, the peptides were treated according to the concentration (0.1 μg / ml, 1 μg / ml and 10 μg / ml), and cultured in 37 ° C medium for 14 days (positive control group: 100 ng / ml tumor necrosis factor-α (Sigma Corporation (SIGMA))). After recovering the cultured cells, ribonucleic acid extraction solution (Easy Blue, Intron) was treated to prepare ribonucleic acid, and complementary deoxyribonucleic acid (cDNA) was synthesized using RT master mix (Intron). A polymerase chain reaction was performed using primers for the respective marker factors (adenylate-activated protein kinase α1, comparative firm gene-58) and a polymerase chain reaction master mix (Intron).
[0099] The target-specific primer sequences for lipogenesis marker fa...
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