GLP-1 derivative
A technology of GLP-1 and derivatives, which is applied in the field of bioengineering, can solve the problems of inactivation and other problems, and achieve the effect of simple production method, mature technology and low cost
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Embodiment 1
[0037] The solid-phase chemical synthesis method synthesizes the GLP-1 derivative of the present invention, and the molecular structure formula of this derivative is: aGLP-1 (7-36), i.e. Seq ID No.2, wherein Xaa2=D-Ala, Xaa20=Ser, Xaa28=Ala, Xaa30=Cys, and the C-terminus of Cys is amide-NH2, operation steps:
[0038] 1. There are 16 amino acid monomers with protective groups, they are: Fmoc-L-Ala-OH, Fmoc-D-Ala-OH, Fmoc-L-His(Trt)-OH, Fmoc-L-Val -OH, Fmoc-L-Glu(OtBu)-OH, Fmoc-L-Tyr(tBu)-OH, Fmoc-L-Gly-OH, Fmoc-L-Leu-OH, Fmoc-L-Thr(tBu)- OH, Fmoc-L-Gln(Trt)-OH, Fmoc-L-Phe-OH, Fmoc-L-Ser(tBu)-OH, Fmoc-L-Ile-OH, Fmoc-L-Asp(OtBu)-OH , Fmoc-L-Trp-OH, Fmoc-L-Cys(Trt)-OH, where the abbreviation means:
[0039] Fmoc: 9-fluorenylmethoxycarbonyl
[0040] Trt: trityl, ie trityl
[0041] OtBu: tert-butyl ester
[0042] tBU: tert-butyl, ie tert-butyl;
[0043] 2. Required equipment and reagents
[0044] Instrument: SYMPHONY 12-channel peptide synthesizer, model: SYMPHONY, American p...
Embodiment 2
[0063] Embodiment 2 The GLP-1 derivative of the present invention is synthesized by solid-phase chemical synthesis. The molecular structural formula of the derivative is aGLP-1(7-37), i.e. Seq ID No.3, wherein Xaa2=D-Ala, Xaa20= Ser, Xaa28=Ala, Xaa30=Gly, Xaa31=Cys, and the C-terminus of Cys is carboxyl-OH, and Wang's resin is selected in this embodiment.
[0064] In the first step of the operation steps, weigh 1 mmol of amino acid monomers with protective groups and bottle them, and arrange them in the SYMPHONY type from the C-terminal to the N-terminal according to the amino acid sequence of the GLP-1 derivative, that is, Seq ID No.3. In the 12-channel peptide synthesizer:
[0065] Fmoc-L-Cys(Trt)-OH, Fmoc-L-Gly-OH, Fmoc-L-Gly-OH, Fmoc-L-Ala-OH, Fmoc-L-Val-OH, Fmoc-L-Leu-OH , Fmoc-L-Trp-OH, Fmoc-L-Ala-OH, Fmoc-L-Ile-OH, Fmoc-L-Phe-OH, Fmoc-L-Glu(OtBu)-OH, Fmoc-L-Ser( tBu)-OH, Fmoc-L-Ala-OH, Fmoc-L-Ala-OH, Fmoc-L-Gln(Trt)-OH, Fmoc-L-Gly-OH, Fmoc-L-Glu(OtBu)-OH , Fmoc-L-Leu...
Embodiment 3
[0067] Example 3 The GLP-1 derivative of the present invention was prepared by solid-phase chemical synthesis. The molecular structural formula of the derivative is aGLP-1(7-38), i.e. Seq ID No.4, wherein Xaa2=Ser, Xaa20=Gln, Xaa28=Asp, Xaa30=Gly, Xaa31=Gly, Xaa32=Cys, and the C-terminus of Cys is carboxyl-OH, Wang's resin is selected in this embodiment.
[0068] In the first step of the operation steps, weigh 1 mmol of the amino acid monomer with the protective group and bottle it, and arrange it in SYMPHONY according to the amino acid sequence of the GLP-1 derivative, namely Seq ID No.4, from the C-terminal to the N-terminal Type 12-channel peptide synthesizer:
[0069]Fmoc-L-Cys(Trt)-OH, Fmoc-L-Gly-OH, Fmoc-L-Gly-OH, Fmoc-L-Gly-OH, Fmoc-L-Asp(OtBu)-OH, Fmoc-L- Val-OH, Fmoc-L-Leu-OH, Fmoc-L-Trp-OH, Fmoc-L-Ala-OH, Fmoc-L-Ile-OH, Fmoc-L-Phe-OH, Fmoc-L-Glu( OtBu)-OH, Fmoc-L-Gln(Trt)-OH, Fmoc-L-Ala-OH, Fmoc-L-Ala-OH, Fmoc-L-Gln(Trt)-OH, Fmoc-L-Gly-OH , Fmoc-L-Glu(OtBu)-OH, Fm...
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