Novel Exenatide Analogs and Their Uses
A technology of exenatide and analogs, applied in the field of new exenatide analogs, can solve the problems of excessive reagents and materials, many reaction steps, short half-life, etc., to achieve excellent blood sugar reduction, prevention or treatment of diabetes, Sustained blood sugar lowering effect
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Embodiment 1
[0129] [Example 1: Preparation method of Ex4(1-32) K-Cap]
[0130] The Ex4(1-32) K-Cap of the present invention comprises a lysine-fatty acid (Lysine-fatty acid) added to the sequence of serine at the 32nd position from the N-terminal of exenatide of the following chemical formula I A peptide of 33 amino acids.
[0131] Chemical formula Ⅰ:
[0132] His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-trp- Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser
[0133] In order to prepare Ex4 (1-32) K-Cap of the present invention, Fmoc-Lys (dde) and dimethylformamide (DMF, Dimethylformamide) are accumulated in trityl resin (trityl resin) to prepare Fmoc-Ser (tBu )-trityl resin. Added N,N-dimethylformamide (N,N-Dimethylformamide; DMF) containing 20% piperidine to the above Fmoc-Ser(tBu)-trityl resin and containing Fmoc-Pro-OH and hydroxybenzene N, N-dimethylformamide of triazole (HOBt, hydroxy-benzotriazole) to prepare Fmoc-Gly-...
Embodiment 2
[0148] [Example 2: Preparation and Effect Analysis of Short Exenatide-4 Analogs]
[0149] [Preparation of short exenatide-4 analogues and stability test against NEP24.11]
[0150] The C-terminus of Exenatide-4 has a 9-AA C-terminal sequence (nine-AAC-terminal sequence) that Glucagon-like Peptide-1 does not have, so it is not easy to decompose into neutral peptides such as NEP24.11 Neutral endopeptidase (reference: Doyle ME et al., Regulatory Peptides Volume 114, Issues 2-3, 15 July 2003, Pages 153-158).
[0151] In order to test the stability of NEP24.11, which is a peptidase, the sequence of the C-terminus of the conventional exendin-4 was changed, and various short exendin-4 (short -exenatide-4) analogs.
[0152] 【(a) Preparation method of Ex4(1-29)】
[0153] To prepare Ex4(1-29), accumulate Fmoc-Gly-OH, N,N-diisopropylethylamine (DIEA, N,N-Diisopropylethylamine) and dimethylformamide in trityl resin to prepare Fmoc -Gly-trityl resin. Add N,N-dimethylformamide containin...
Embodiment 3
[0197] [Example 3: Characteristic Analysis of Ex4(1-32) K(Cap)]
[0198] 【Ex4(1-32)K(Cap)(subcutaneous) glucose tolerance test using diabetic model mice】
[0199] After making male db / db mice (6-week-old to 12-week-old) fast for 18 hours, feed them to fasting mice at concentrations of 0.005 nmol / kg, 0.01 nmol / kg, 0.1 nmol / kg, 1 nmol / kg and 5 nmol / kg, respectively. The diabetic model mice were administered (subcutaneously) short exenatide-fatty acid conjugated Ex4(1-32)K(Cap), and 30 minutes later, glucose (1.5 g / kg) was intraperitoneally administered. After 0 minutes, 20 minutes, 40 minutes, 60 minutes, 90 minutes, 120 minutes, and 180 minutes, blood was collected from the tail end of the mice, and blood glucose was measured with a blood glucose meter. As a result, the concentration-dependent hypoglycemic effect of Ex4(1-32)K(Cap) was confirmed ( Figure 13a and Figure 13b ).
[0200] For comparison with Exenatide as a positive control group, Ex4(1-32)K(Cap) and Exenatide...
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