Total synthesis and structural elucidation of azaspiracid-1. Final assignment and total synthesis of the correct structure of azaspiracid-1.

The molecular structure of azaspiracid-1, a neurotoxin isolated from mussels, has been elucidated by total synthesis which also enriched its supplies. The degradatively derived fragments of this marine biotoxin, compounds 5 (EFGHI), 6 (FGHI), and 40 (ABCD), were matched with synthetic materials, thus confirming their structural identities. Based on this detective work, a new structure of azaspiracid-1 (i.e., 1) was proposed and constructed by total synthesis. The final strategy for the total synthesis of azaspiracid-1 featured a dithiane anion (C(21)-C(27) fragment) reacting with a pentafluorophenol ester (C(1)-C(20) fragment) followed by a Stille-type union of an advanced allylic acetate substrate (C(1)-C(27) fragment) with a vinyl stannane as the main coupling processes to assemble the carbon skeleton of the molecule. In addition to the total synthesis of azaspiracid-1 (1), the syntheses of its C(1)-C(20) epimer (2) and of several truncated analogues for biological investigations are described.

The effects of acute exercise on serum adiponectin and resistin levels and their relation to insulin sensitivity in overweight males.

The purpose of this study was to investigate the effects of a submaximal aerobic exercise bout on adiponectin and resistin levels as well as insulin sensitivity, until 48 h post-exercise in healthy overweight males. Nine subjects performed an exercise bout at an intensity corresponding to approximately 65% of their maximal oxygen consumption for 45 min. Adiponectin, resistin, cortisol, insulin, glucose and insulin sensitivity were measured prior to exercise, immediately after exercise as well as 24 and 48 h after exercise. Data were analyzed using repeated measures ANOVA while Pearson's correlations were performed to identify possible relationship among the assessed variables. There were no significant differences for adiponectin (microg ml(-1)) [pre, 3.61(0.73); post, 3.15(0.43); 24 h, 3.15(0.81); 48 h, 3.37(0.76)] or resistin (ng ml(-1)) [pre, 0.19(0.03); post, 0.13(0.03); 24 h, 0.23(0.04); 48 h, 0.23(0.03)] across time. Insulin sensitivity increased and insulin concentration decreased significantly only immediately after exercise. Furthermore, no significant correlations were observed among the variables assessed except for the expected between insulin level and insulin sensitivity. These results indicate that a submaximal aerobic workout does not result in significant changes in adiponectin and resistin up to 48 h post-exercise. Furthermore, it appears that adiponectin or resistin is not associated with insulin sensitivity.

Total synthesis and structural elucidation of azaspiracid-1. Construction of key building blocks for originally proposed structure.

Syntheses of the three key building blocks (65, 98, and 100) required for the total synthesis of the proposed structure of azaspiracid-1 (1a) are described. Key steps include a TMSOTf-induced ring-closing cascade to form the ABC rings of tetracycle 65, a neodymium-catalyzed internal aminal formation for the construction of intermediate 98, and a Nozaki-Hiyama-Kishi coupling to assemble the required carbon chain of fragment 100. The synthesized fragments, obtained stereoselectively in both their enantiomeric forms, were expected to allow for the construction of all four stereoisomers proposed as possible structures of azaspiracid-1 (1a-d), thus allowing the determination of both the relative and absolute stereochemistry of the natural product.

Total synthesis and structural elucidation of azaspiracid-1. Synthesis-based analysis of originally proposed structures and indication of their non-identity to the natural product.

The key building blocks (6, 7, and 8) for the intended construction of the originally proposed structures of azaspiracid-1, a potent marine-derived neurotoxin, were coupled and the products elaborated to the targeted compounds (1a,b) and their C-20 epimers (2 and 3). The assembly of the three intermediates was accomplished by a dithiane-based coupling reaction that united the C(1)-C(20) (7) and C(21)-C(27) (8) fragments, followed by a Stille-type coupling which allowed the incorporation of the C(28)-C(40) fragment (6) into the growing substrate. Neither of the final products (1a,b) matched the natural substance by TLC or (1)H NMR spectroscopic analysis, suggesting one or more errors in the originally proposed structure for this notorious biotoxin.

Second-generation total synthesis of azaspiracids-1, -2, and -3.

The naturally occurring but scarce marine neurotoxins azaspiracids-1, -2, and -3 have been synthesized from five key building blocks by a convergent strategy that involved dithiane and Stille coupling reactions. The ABCD fragments were constructed through a cascade reaction involving deprotection/self-assembly of the precursors, while the FGHI fragment was forged by a neodymium triflate-induced cyclization. The final ring closure (ring G) was achieved, after the union of all fragments, through an iodoetherification reaction followed by reductive removal of the facilitating iodine residue. These improved, second-generation routes confirm the absolute structures and render all three azaspiracids readily available for biological studies.

Comparison between leg and arm eccentric exercises of the same relative intensity on indices of muscle damage.

Many exercise models have demonstrated associations between eccentric muscle actions and muscle damage. However, the magnitude of muscle damage varies among the models. It appears that responses to eccentric exercise are different between leg and arm muscles but this has not been systematically clarified. This study compared leg and arm eccentric exercises of the same relative intensity for indices of muscle damage. Eleven healthy untrained males [Age: 21.2 (1.0) years, Height: 179.4 (3.0) cm, Weight: 78.4 (3.1) kg] performed a sub-maximal eccentric exercise of the knee extensors (LEGS) and the elbow flexors (ARMS), separately. Both LEGS and ARMS consisted of six sets of 12 repetitions with an intensity corresponding to 75% of the predetermined maximal eccentric peak torque (EPT) of each muscle. Range of motion (ROM), delayed onset muscle soreness (DOMS), serum creatine kinase (CK) and lactate dehydrogenase (LDH) activities, myoglobin (Mb) concentration, and muscle strength [EPT and isometric peak torque (IPT)] were assessed before and 24, 48, 72, and 96 h following exercise. Significant (P < 0.05) changes in DOMS and ROM were observed up to 96 h after both exercise bouts, and the magnitude of the change was similar between LEGS and ARMS. Increases in CK and Mb were significantly (P < 0.05) larger after ARMS than LEGS at 72 and 96 h post-exercise. EPT and IPT were significantly (P < 0.05) lower than the baseline up to 96 h post-exercise for ARMS but were fully recovered by 96 h post-exercise for LEGS. Decreases in muscle strength were significantly (p < 0.05) larger following ARMS than LEGS at 48, 72, and 96 h post-exercise for EPT, and from 24 h to 96 h post-exercise for IPT. These results suggest that the magnitude of muscle damage is greater and the recovery of muscle function was slower after eccentric exercise of arm elbow flexors than the knee extensors.