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To this end, while a total of 564 amino acid sequences (supplementary file rewriteEverything.out ) of semaglutide backbone were designed with the sequence template in Figure 1, the 564 site-specific missense mutations were screened from a list of g(28,7) (Table 6) semaglutide analogue canditates whose GLP-1R ECD affinity first reached as strong as 1.0 × 10 −9 M, i.e., K d as low as 1.0 × 10 −9 M or lower (raw data not published)