Ryanodine (1) is a plant-derived natural product with powerful pharmacological and insecticidal action, and is a potent modulator of intracellular calcium release channels. Compound 1 possesses a 1H-pyrrole-2-carboxylate ester at the C3-position of heptahydroxylated terpenoid ryanodol (2). Whereas 2 was readily obtained from 1 by basic hydrolysis, 1 has never been synthesized from 2, due to the extreme difficulty in selectively introducing the bulky pyrrole moiety at the severely hindered C3-hydroxyl group of heptaol 2. Here we report chemical conversion of 2 to 1 for the first time. The derivatization was realized through the use of a new protective group strategy and the application of on-site construction of the pyrrole-2-carboxylate ester from the glycine ester and 1,3-bis(dimethylamino)allylium tetrafluoroborate.Key words acylation; protecting group; regioselectivity; ryanodine; terpenoid Ryanodine (1, Chart 1) has been isolated from the wood of Ryania speciosa VAHL and shown to possess potent insecticidal and pharmacological actions.1) Compound 1 binds specifically to a high-conductance intracellular calcium channel known as the ryanodine receptor (RyR), altering its conductance.2,3) Since the RyR plays an important role in many biological processes and malfunctions in its action have been linked to skeletal and cardiac diseases, 4-7) 1 has been used as a biological probe for investigations of RyR function, and is regarded as a strong candidate in the development of therapeutic agents for various diseases. [8][9][10] In 1968, the structure of ryanodine (1) was determined to be the ester between 1H-pyrrole-2-carboxylic acid (3) and the C3-hydroxyl group of ryanodol (2). [11][12][13][14] Compound 2 is a diterpenoid containing the exceedingly complex five fused rings (ABCDE-rings) with eleven contiguous stereocenters. Interestingly, the affinity of 2 toward RyRs of vertebrate skeletal muscle is 1700 times weaker than that of 1 (K D =7 nM), [15][16][17] demonstrating that the pyrrole group is a primary determinant for strong interactions of 1. Because of its important biological activities and intriguing structural features, many studies attempting the chemical construction of 1 has been carried out, which resulted in the total syntheses of ryanodol (2) by the Deslongchamps group in 1979 18,19) and by our laboratory in 2014. [20][21][22] In 1951, before the complete structure of 1 was known, Wiesner and colleagues found that basic hydrolysis of ryanodine (1) generated 2 and 3.11) However, the reverse reaction has not been realized even to date. The difficulty of the transformation arises from the need to control the regioselectivity of the four tertiary (C2, 4, 6, 12), two secondary (C3, 10) and one acetalic (C15) alcohols within heptaol 2 and to accommodate the bulky pyrrole moiety at the hindered C3-hydroxyl group. The computer-generated 23) structure of 2 revealed that its axially oriented C3-OH was shielded by the proximal C14-hydrogen atom and by the C19-, 20-methyl groups (Fig. 1). In fact, direct esteri...