Background Technology
Cagrilintide is a novel long-acting acylated amylin analog. As a non-selective agonist of the amylin receptor (AMYR) and the calcitonin G protein-coupled receptor (CTR), it is a cyclic polypeptide composed of 38 amino acids. Its sequence contains 38 amino acids and a pair of disulfide bonds. Cagrilintide can significantly reduce body weight and food intake, and has potential in obesity research. Amylin is another hormone related to hunger and satiety outside the GLP-1 signaling pathway.
Cagrilintide can reduce energy intake, regulate food choices and preferences, exert a glucose-regulating effect by being co-secreted with insulin, inhibit postprandial glucagon release, and delay gastric emptying. It has obvious advantages in the treatment of diabetes, obesity, metabolic syndrome, cardiovascular diseases and other aspects, and has broad demand and application prospects.
Chemical Structural Formula of Cagrilintide
Basic Information
English Name：Cagrilintide
Company Number：GT-F064
CAS：1415456-99-3
Sequence：γ-Glu-Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Arg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-Ile-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Pro-NH2(Disulfidebridge:cys4-cys9)
Molecular Formula：C194H312N54O59S2
Molecular Weight：4409.01
Raw materials used for Cagrilintide
Synthetic Route
Cagrilintide Preparation Diagram
(1)Preparation of Cagrilintide Peptide Resin
Take 1.5 mmol of the first protected amino acid and 1.5 mmol of HOBT, dissolve with an appropriate amount of DMF and cool to 0~15°C; take another 1.5 mmol of DIC, slowly add it to the DMF solution of the protected amino acid under stirring, and stir the reaction for 10 minutes at 0~15°C to obtain an activated protected amino acid solution, which is set aside.
Take 0.5 mmol of amino resin (substitution degree 0.42 mmol/g), swell it with DMF solution for 60 minutes, deprotect it with 20% Pip/DMF solution for 30 min, wash and filter to obtain the resin to be reacted.
Add the activated first protected amino acid solution to the resin to be reacted, carry out the coupling reaction at 25~35℃ for 120~300 minutes, filter and wash to obtain the resin containing 1 protected amino acid: Fmoc-Pro-Resin
Using the same method as above, sequentially access the corresponding 2nd to 38th protected amino acids or fragments mentioned above:Fmoc- Thr(tBu)-OH，Fmoc-Asn(Trt)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Gly-OH，Fmoc-Val-OH，Fmoc-Asn(Trt)-OH，Fmoc-Thr(tBu)-OH，Fmoc-Pro-OH·H2O，Fmoc-Pro-OH·H2O，Fmoc-Leu-OH，Fmoc-Ile-OH，Fmoc-Pro-OH·H2O，Fmoc-Phe-OH，Fmoc-Asn(Trt)-OH，Fmoc-Asn(Trt)-OH，Fmoc-Ser(tBu)-OH，Fmoc-Ser(tBu)-OH，Fmoc-His(Trt)-OH，Fmoc-Arg(Pbf)-OH，Fmoc-Leu-OH，Fmoc-Pro-OH·H2O，Fmoc-Glu(OtBu)-OH，Fmoc-Ala-OH·H2O，Fmoc-Leu-OH，Fmoc-Arg(Pbf)-OH，Fmoc-Gln(Trt)-OH，Fmoc-Ala-Thr(psi(Me,Me)pro)-OH，Fmoc-Cys(Trt)-OH，Fmoc-Ala-Thr(psi(Me,Me)pro)-OH，Fmoc-Thr(tBu)-OH，Fmoc-Asn(Trt)-OH，Fmoc-Cys(Trt)-OH，Fmoc-Lys(Boc)-OH，Fmoc-Glu-OtBu，Mono-tert-butyl eicosanedioate, yielding 4.98 g of carmegliptin peptide resin
(2)Preparation of crude Cagrilintide linear peptide
Take 4.49 g of carfilzomib peptide resin, add TFA mixed solution, the ratio of the mixed solution is TFA:EDT/Tis/phenol/H₂O = 87.5:2.5:5:2.5:2.5, and the dosage is 8 ml per gram of resin; stir the reaction at 20~30°C for 3 hours, filter the reaction mixture with a sand core funnel, collect the filtrate, wash the resin with a small amount of TFA 3 times, combine the filtrates, add MTBE for precipitation, wash the precipitate with MTBE 3 times, and pump dry to obtain an off-white powder, which is 2.08 g of crude linear carfilzomib peptide.
(3)Preparation of crude Cagrilintide cyclic peptide solution
Take 0.50 g of crude linear Cagrilintide peptide, prepare a crude linear peptide solution at a concentration of 5.0 mg/ml, add 5 ml of DMSO solution dropwise under stirring, and react at 20~30℃ for 20 h to obtain a crude cyclic cagrilintide peptide solution.
(4)Preparation of refined Cagrilintide peptide
Take the crude Carraglitide cyclic peptide solution, and filter it with a 0.45μm mixed microporous filter membrane；
Purify and prepare using high performance liquid chromatography, and collect the Carragliptin fractions；
High-performance liquid chromatography was used for buffer exchange. The main peak after buffer exchange was collected and its purity was detected by analytical liquid chromatography. The main peak solutions after buffer exchange were combined, concentrated under reduced pressure to obtain an aqueous acetic acid solution of carbotide, which was freeze-dried to obtain 152.18 mg of refined carbotide with a purity of 99.58%, a maximum single impurity of 0.09%, a total yield of 31.87%, and a molecular weight of 4409.0
HPLC diagram
MS diagram
The above examples show that, compared with the step-by-step coupling synthesis process, the product obtained by the method provided by the present invention has a purity of more than 99.0%, and each single impurity is less than 0.1%, which improves the product quality and total yield. The process is simple and easy to control, has a high degree of industrialization, and has extensive practical value and application prospects.
Contact Us
Phone: +86-18986204913
Tel: +86-027-83338163
E-Mail: info@cosperpharma.com
