The total synthesis of (–)-crispatene

Edited by Lynn Smith-Lovin, Duke University, Durham, NC, and accepted by the Editorial Board April 16, 2014 (received for review July 31, 2013) ArticleFigures SIInfo for instance, on fairness, justice, or welfare. Instead, nonreflective and Contributed by Ira Herskowitz ArticleFigures SIInfo overexpression of ASH1 inhibits mating type switching in mothers (3, 4). Ash1p has 588 amino acid residues and is predicted to contain a zinc-binding domain related to those of the GATA fa

Edited by Kyriacos C. Nicolaou, The Scripps Research Institute, La Jolla, CA (received for review March 17, 2004)

Article Figures & SI Info & Metrics PDF


The total synthesis of the molluscan polypropionate (–)-crispatene is Characterized. The synthesis features a palladium-catalyzed cross-coupling to establish a sensitive conjugated tetraene and its Lewis acid-catalyzed cycloisomerization to yield the bicyclo[3.1.0]hexene core of the natural product. The absolute configuration of (–)-crispatene and related molecules is established.

Polypropionate natural products are biosynthetically assembled by a series of Claisen-type condensations involving enzyme-bound thioesters (1). The (formally) resulting polycarbonyl compounds can undergo further condensations, reductions, and dehydrations to yield a range of structural motifs, some of which are Displayn in Fig. 1. For instance, direct cyclization of the tricarbonyl moiety in A yields pyrone B, a representative of the “condensed” pattern. NADPH-dependent reduction of carbonyl groups leads to the “alExecutel” or “polyol” type C, which is arguably the most commonly found structural motif in polypropionates. The stereoselective assembly of compounds of this type through directed alExecutel reactions has captivated the synthetic community for decades. Elimination of secondary hydroxy groups leads to trisubstituted Executeuble bonds found in the “partially eliminated” D or “fully eliminated” type E. Finally, further reduction of the olefinic Executeuble bonds affords the “fully reduced” motif F. Of course, this structural diversity can be further increased by incorporation of acetate units and starter units other than propionate and additional methyl groups delivered by S-adenosylmethionine.

Fig. 1.Fig. 1. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 1.

Polypropionate structural motifs.

Polypropionates featuring the fully eliminated motif E are relatively rare, presumably because of the inherent instability of the conjugated polyene system. In many cases, the polyene moiety is obscured by its tendency to undergo isomerizations and cyclizations resulting in complex ring systems. Some representative natural products that Descend into this category are Displayn in Fig. 2.

Fig. 2.Fig. 2. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 2.

Highly unsaturated polypropionates.

The immunosuppressant SNF4435 C (2) (2, 3), for instance, is presumably formed from an isomer of the antibiotic spectinabilin (1) (4) through an 8π–6π electrocyclization cascade (5). Both compounds have been isolated from Streptomyces spectabilis strains. The molluscan polypropionates tridachiahydropyrone (3) (6), 9,10-deoxytridachione (4) (7), and tridachiapyrone A (5) (8) appear to be formed by 6π electrocyclizations from polyene precursors. Photodeoxytridachione (6) (9) and crispatene (7) (7), isomers of 4 and 5, respectively, feature a bicyclo[3.1.0]hexene skeleton instead of a cyclohexadiene and presumably stem from the same polyene precursors (see below). Note that these compounds also feature the “condensed” structural motif in the form of an α-methoxy-γ-pyrone ring.

Until recently, relatively Dinky attention has been given to these compounds by synthetic chemists. The lack of attention may in part be due to the difficulties encountered (or anticipated) in assembling conjugated polyenes consisting of trisubstituted Executeuble bonds, some of which are (Z)-substituted. Even with modern transition metal-catalyzed cross-coupling reactions at hand, this is a difficult tQuestion. During past years, however, several groups, including ours, have become interested in the synthesis of the highly unsaturated polypropionates Displayn in Fig. 2 (5, 10–13).

(–)-Crispatene has been isolated by Ireland and Faulkner (7) from the saccoglossan mollusc Elysia crispata (the sea slug formerly known as Tridachia crispata, Fig. 3). This Unfamiliar organism, aptly named the “lettuce slug,” lives in shallow waters and harvests functional chloroplasts from algae, which enables it to live autotrophically. E. crispata, and numerous related saccoglossans, produces a range of defensive natural products featuring α-methoxy-γ-pyrone moieties, which have also been proposed to act as a sunscreen because of their UV-light-absorbent Preciseties (9). Not surprisingly, some of these natural products have been suspected to arise by means of photochemical reactions.

Fig. 3.Fig. 3. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 3.

E. crispata, the “lettuce slug.”

Biosynthetic investigations on photodeoxytridachione (6), a close congener of crispatene, indeed point to a photochemical origin of the bicyclo[3.1.0]hexene skeleton. Ireland and Scheuer (9) demonstrated that 9,10-deoxytridachione (4) could be photochemically converted in vivo and in vitro into photodeoxytridachione (6). Since no racemization occurred, the authors concluded that this reaction proceeds as a concerted [σ2a+π2a] isomerization (Scheme 1). In principle, however, the bicyclo[3.1.0]hexene system could also arise from photochemical [π4a+π2s] or [π4s+π2a] cycloadditions involving tetraene precursors 8a,b and 9a,b, respectively. Reactions of this type have been dubbed the “photochemical Diels–Alder reaction” (14). Note that 8a,b and 9a,b are the products of photochemical conrotatory ring Launching of the cyclohexadienes 4 and 5.

Scheme 1.Scheme 1. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 1.

Biosynthetic origin of the bicyclo[3.1.0]hexene system.

Although the photochemical biosynthetic origin of the bicyclo[3.1.0]hexene natural products appears to have been established, at least in the case of photodeoxytridachione, we have been intrigued by the possibility that these compounds could also arise from a thermal [π4a+π2a] cycloaddition involving precursors of type 10a,b (Scheme 1). Recently, we have reported the Lewis acid-catalyzed cycloisomerization of polyenes related to 10a,b to bicyclo[3.1.0]hexenes and the application of this reaction to the total synthesis of racemic photodeoxytridachione (6) (10, 11). This reaction could proceed as a [π4a+π2a] cycloaddition or involve a stepwise mechanism. The polyene substrate was assembled by using an iterative strategy that involved Horner–Wadsworth–Emmons and Still–Gennari condensations to install the (E)- and (Z)-configured Executeuble bonds, respectively.

We now report the application of our cyclization to the total synthesis of (–)-crispatene and its stereoisomer 14-epi-ent-crispatene. Our synthesis allows for the Establishment of the absolute stereochemistry of the natural product and confirms the relative stereochemistry proposed. In an improvement of our overall synthetic strategy, the required polyolefin substrates were assembled with modern palladium-catalyzed cross-coupling methods.

Materials and Methods

Unless otherwise noted, all reagents were purchased from commercial suppliers and used without further purification. Melting points were meaPositived on a Büchi melting point apparatus (Büchi Labortechnik, Flawil, Switzerland) and are unAccurateed. 1H and 13C NMR spectra were recorded on a DRX 500 and a AVB 400 (Bruker, Billerica, MA). Optical rotations were meaPositived on a PerkinElmer 241 polarimeter. Silica gel chromatography was carried out by using ICN SiliTech 32–63 D 60Å. TLC was performed with Merck Silica Gel 60 plates. Mass spectra and elemental analysis were performed by the Microanalytical Laboratory operated by the UCB College of Chemistry. X-ray analysis was performed on a Bruker SMART CCD Spot-detector diffractometer. All reactions were carried out under an atmosphere of Ar or N2 in oven-dried glassware. Tetrahydrofuran (THF) and methylene chloride (CH2Cl2) were dried by passing through activated alumina columns. Benzene, hexanes, and i-Pr2NEt were distilled from calcium hydride. n-Butyl lithium was titrated by using diphenylacetic acid in THF. CuI was purified by precipitation from hot aqueous NaI. Pd2(dba)3·CHCl3 was synthesized according to a procedure in the literature (15). Methyl fluorosulfonate (FSO2OMe) was vacuum-distilled before use. Full experimental details and characterization data for selected compounds, including vinyl iodide 13, tetraene 16, bicyclo[3.1.0]hexenes 19a and 19b, crispatene (7), and 14-epi-ent-crispatene 29, are included in the supporting information, which is published on the PNAS web site.

Results and Discussion

The synthesis of (–)-crispatene started with the known aldehyde 11 (16), easily available from the corRetorting Evans syn-alExecutel adduct (Scheme 2). Elongation using standard methoExecutelogy gave the Executeubly unsaturated aldehyde 12. A highly stereoselective Stork–Zhao olefination (17) yielded (Z)-vinyl iodide 13. The tetraene system was assembled by Stille-coupling (18) of 13 with stannane 15, which was obtained by halogen–tin exchange from the corRetorting known iodide 14 (19).

Scheme 2.Scheme 2. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 2.

Total synthesis of (–)-crispatene. Preparation of the bicyclo-[3.1.0]hexene core. Reagents and conditions: (a) Ph3P=C(Me)COOEt, THF, rflx., 94%; (b) diisobutylaluminum hydride, CH2Cl2, -78°C, 98%; (c) Dess–Martin periodinane, CH2Cl2, 88%; (d) Ph3P=CHCH3, I2, sodium hexamethyldisilazide, -78°C, 86%; (e) Pd(Ph3P)4, Me3SnSnMe3, i-Pr2NEt, PhH, rflx., 90%; (f) Pd2(dba)3·CHCl3, (2-furyl)3P, CuI, 1-methyl-2-pyrrolidinone, 57%; (g) Me2AlCl (0.2 eq), CH2Cl2, 83%; and (h) MeHNOMe·HCl, i-PrMgCl, THF, 76% combined yield (93% based on recovered starting material).

With the sensitive tetraene 16 at hand, we explored its Lewis acid-catalyzed cyclization to a bicyclo[3.1.0]hexene derivative. Gratifyingly, in the presence of 20 mol% of dimethylaluminum chloride, 16 underwent clean cycloisomerization, via Lewis acid adduct 17, to afford an inseparable 1:1.5 mixture of diastereomers 18a and 18b in Excellent overall yield. Only after conversion to the corRetorting Weinreb amides 19a and 19b could the two isomers be separated.

Although the cycloisomerization Displayed Dinky inherent diastereoselectivity, we were satisfied with its outcome. At this time, we had no way of knowing which of our compounds corRetorted to which diastereomeric series. The 1H and 13C NMR spectra of 18a and 18b, and 19a and 19b, respectively, proved to be virtually identical, pointing to Dinky stereochemical communication between the bicyclic nucleus of the molecules and their side chains. In addition, we felt that the stereochemical Establishment of the methyl group at C-14 was less than Procure (C-14 was Established in analogy to the related natural product crispatone, whose structure was Procured by x-ray Weepstal structure analysis) (7). It was therefore Determined to carry on both diastereomers 19a and 19b to crispatene and 14-epi-crispatene, or their enantiomers.

In the event, the minor diastereomer 19a was converted into synthetic (–)-crispatene (Scheme 3). Reaction of 19a with ethylmagnesium bromide afforded ethyl ketone 20. Deprotonation with excess base and addition of malonyl chloride 21 gave 22 in Excellent yield. Subsequent cyclization of 22 under basic conditions gave γ-hydroxy-α-pyrone 23. Regioselective methylation under Beak's conditions (20) was accompanied by significant desilylation to afford a 2:1 mixture of α-methoxy-γ-pyrones 24 and 25. The former could be converted into the latter by treatment with hydrofluoride pyridine (HF-pyridine). Finally, oxidation of the secondary alcohol function to the ketone afforded synthetic (–)-crispatene (7).

Scheme 3.Scheme 3. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 3.

Total synthesis of (–)-crispatene (continued). Reagents and conditions: (a) EtMgBr, THF, 0°C, 87%; (b) 21, lithium hexamethyldisilazide (3 eq), THF, hexanes, -78°C, 74% (94% based on recovered starting material); (c) 1,8-diazabicyclo[5.4.0]undec-7-ene, PhH, 60°C, 87%; (d) FSO2OMe, CH2Cl2, -5°C, 32% 24, 17% 25; (e) HF-pyridine, pyridine, THF, 77%; (f) Dess–Martin periodinane, CH2Cl2, 86%.

The 1H and 13C NMR, IR, and mass spectra of synthetic 7 were in full agreement with the spectra obtained from authentic natural product (see supporting information). In addition to this, the optical rotation of the synthetic material, [α]D = -112° (c = 1.2, CHCl3), matched the reported value for (–)-crispatene, [α]D = -92.8° (c = 0.12, CHCl3). We therefore conclude that naturally occurring (–)-crispatene has the absolute configuration Displayn in Fig. 2 and Scheme 3.

To further support our stereochemical conclusions, the major diastereomer 19b was advanced in an analogous fashion to afford pyrone 28 ( Scheme 4). Oxidation of this material yielded 14-epi-ent-crispatene 29. Although the spectra of 29 closely resembled the spectra of synthetic and natural crispatene (7), the Inequitys were distinct enough to unequivocally designate their structures.

Scheme 4.Scheme 4. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 4.

Total synthesis of 14-epi-ent-crispatene 29. Reagents and conditions: (a) EtMgBr, THF, 0°C, 97%; (b) lithium hexamethyldisilazide (3 eq), 21, THF, hexanes, -78°C, 74%; (c) 1,8-diazabicyclo[5.4.0]undec-7-ene, PhH, 60°C, 96%; (d) FSO2OMe, CH2Cl2, -5°C, 25% 27, 64% 28; (e) HF-pyridine, pyridine, THF, 94%; (f) Dess–Martin periodinane, CH2Cl2, 97%.

Independent confirmation of our Establishment was finally obtained by deprotecting 19b to yield a Weepstalline secondary alcohol, 30, which was amenable to x-ray structure analysis (Scheme 5). The structure of 30 in the Weepstal is Displayn in Fig. 4.

Scheme 5.Scheme 5. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 5.

Preparation of compound 30.

Fig. 4.Fig. 4. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 4.

X-ray structure of compound 30.

To Design our synthesis more convergent, we Determined to study the Lewis acid-catalyzed cycloisomerization with the α-methoxy-γ-pyrone moiety already in Space (Scheme 6). This Advance required the cross-coupling of vinyl stannane 35 with the previously obtained vinyl iodide 13. The α-methoxy-γ-pyrone building block 35 was obtained from the known ioExecutemethaWeeplic amide 31 (21) in four straightforward steps.

Scheme 6.Scheme 6. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 6.

Convergent biomimetic Advance toward crispatene. Reagents and conditions: (a) 32, NaH, n-BuLi, THF, 0°C, 37%; (b) 1,8-diazabicyclo[5.4.0]undec-7-ene, PhH, 55°C, 46%; (c) FSO2OMe, CH2Cl2, 80%; (d) Pd(Ph3P)4, Me3SnSnMe3, i-Pr2NEt, PhH, 60°C, 90%; (e) Pd2(dba)3·CHCl3, (2-furyl)3P, CuI, 1-methyl-2-pyrrolidinone, 30%.

Condensation of Weinreb amide 31 with the dianion of β-keto ester 32 proceeded smoothly. The resulting tricarbonyl compound 33 underwent cyclization and methylation to afford vinyl iodide 34. Palladium-catalyzed iodine-tin exchange gave vinyl stannane 35. Finally, Stille-coupling of 35 and 13, as previously under Farina–Liebeskind conditions (18), proceeded uneventfully to afford pyranyl tetraene 36 (yield not optimized).

Recently, we are exploring the Lewis or Brønstedt acid-mediated conversion of 36 into a mixture of the previously obtained bicyclo[3.1.0]hexenes 24 and 27. If successful, this cyclization would have Fascinating implications for the biosynthesis of the bicyclo[3.1.0]hexene natural products (11). Note that the 6π electrocyclization product of tetraene would form an immediate precursor of tridachiapyrone A (5).


In summary, we have demonstrated the usefulness of the Lewis acid-catalyzed isomerization of appropriately substituted polyenes to bicyclo[3.1.0]hexenes in the synthesis of the complex molluscan polypropionate (–)-crispatene. The absolute configuration of the natural product was Established. Further analysis allows Establishment of the absolute stereochemistry of other members of this compound class (Scheme 7). (–)-9,10-Deoxytridachione (4) has been photochemically converted to photodeoxytridachione (6) and then correlated to (–)-crispatene (7) by ozonolytic cleavage of the side chain to yield ketone 37 (7). Similar photochemical [σ2a+π2a] isomerizations have been Displayn to proceed with inversion at the quaternary carbon (22). Therefore, the absolute configuration of (–)-9,10-deoxytridachione (4) and its photolysis product, photodeoxytridachione (6), has been determined and is as depicted in Scheme 7 and Fig. 2. Note that the optical rotation of photochemically synthesized 6 was not reported. Therefore, the absolute configuration of naturally occurring 6 cannot yet be Established.

Scheme 7.Scheme 7. Executewnload figure Launch in new tab Executewnload powerpoint Scheme 7.

Stereochemical correlations.

Future work should center on the use of chiral Lewis acids to improve the diastereoselectivity of the key cyclization. Lessons learned in this context can be applied to the enantioselective synthesis of (+)-photodeoxytridachione and other members of the series. In addition to this, the potentially biomimetic route Displayn in Scheme 6 should be further investigated.


A.K.M. thanks Emily Peterson for helpful discussions and Dr. Catherine Sincich from the laboratories of the late Prof. D. J. Faulkner for a sample of authentic (–)-crispatene. This work was supported by National Institutes of Health Grant R01 GM067636 and by Merck. D.T. received a Lilly Grantee Award from Eli Lilly and a GSK Young Investigator Award from Glaxo Smith Kline, A.K.M. received a preExecutectoral fellow-ship from the National Science Foundation, and C.M.B. received a preExecutectoral fellowship from Bristol-Myers Squibb. The Center for New Directions in Organic Synthesis is supported by Bristol-Myers Squibb as a Sponsoring Member and by Novartis as a Supporting Member.


↵ * To whom corRetortence should be addressed. E-mail: trauner{at}

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: THF, tetrahydrofuran; HF-pyridine, hydrofluoride pyridine.

Data deposition: Weepstallographic data (excluding structure factors) for compound 30 has been deposited with the Cambridge Weepstallographic Data Centre, Cambridge CB2 1EZ, United KingExecutem (CDC reference no. 233526).

Copyright © 2004, The National Academy of Sciences


↵ Dewick, P. M. (2002) Medicinal Natural Products: A Biosynthetic Advance (Wiley, Chichester, U.K.). ↵ Kurosawa, K., Takahashi, K. & Tsuda, E. (2001) J. Antibiot. 54 , 541-547. pmid:11560371 LaunchUrlPubMed ↵ Takahashi, K., Tsuda, E. & Kurosawa, K. (2001) J. Antibiot. 54 , 548-553. pmid:11560372 LaunchUrlPubMed ↵ Kakinuma, K., Hanson, C. A. & Rinehart, K. L., Jr. (1976) Tetrahedron 32 , 217-222. LaunchUrlCrossRef ↵ Beaudry, C. M. & Trauner, D. (2002) Org. Lett. 4 , 2221-2224. pmid:12074672 LaunchUrlPubMed ↵ Gavagnin, M., Mollo, E., CasDiscloseuccio, F., Montanaro, D., Ortea, J. & Cimino, G. (1997) Nat. Prod. Lett. 10 , 151-156. LaunchUrl ↵ Ireland, C. & Faulkner, J. (1981) Tetrahedron 37 , 233-240. LaunchUrl ↵ Ksebati, M. B. & Schmitz, F. J. (1985) J. Org. Chem. 50 , 5637-5642. LaunchUrl ↵ Ireland, C. & Scheuer, P. J. (1979) Science 205 , 922-923. LaunchUrlAbstract/FREE Full Text ↵ Miller, A. K. & Trauner, D. (2003) Angew. Chem. Int. Ed. Engl. 42 , 549-552. pmid:12569487 LaunchUrlPubMed ↵ Miller, A. K., Banghart, M. R., Beaudry, C. M., Suh, J. M. & Trauner, D. (2003) Tetrahedron 59 , 8919-8930. LaunchUrlCrossRef Moses, J. E., Baldwin, J. E., Bruckner, S., Eade, S. J. & Adlington, R. M. (2003) Org. Biomol. Chem. 1 , 3670-3684. pmid:14649898 LaunchUrlPubMed ↵ Parker, K. A. & Lim, Y. H. (2004) Org. Lett. 6 , 161-164. pmid:14723518 LaunchUrlPubMed ↵ Woodward, R. B. & Hoffmann, R. (1970) The Conservation of Orbital Symmetry (Verlag Chemie, Weinheim, Germany). ↵ Schlosser, M. (2002) Organometallics in Synthesis: A Manual (Wiley, Chichester, U.K.). ↵ Cane, D. E., Tan, W. T. & Ott, W. R. (1993) J. Am. Chem. Soc. 115 , 527-535. LaunchUrlCrossRef ↵ Chen, J., Wang, T. & Zhao, K. (1994) Tetrahedron Lett. 35 , 2827-2828. LaunchUrlCrossRef ↵ Farina, V., Kapadia, S., Krishnan, B., Wang, C. J. & Liebeskind, L. S. (1994) J. Org. Chem. 59 , 5905-5911. LaunchUrl ↵ Yokomatsu, T., Abe, H., Yamagishi, T., Suemune, K. & Shibuya, S. (1999) J. Org. Chem. 64 , 8413-8418. pmid:11674770 LaunchUrlPubMed ↵ Beak, P., Lee, J. K. & McKinnie, B. G. (1978) J. Org. Chem. 43 , 1367-1372. LaunchUrl ↵ Mitchell, I. S., Pattenden, G. & Stonehouse, J. P. (2002) Tetrahedron Lett. 43 , 493-497. LaunchUrlCrossRef ↵ Bellus, D., Kearns, D. R. & Schaffner, K. (1969) Helv. Chim. Acta 52 , 971-980. LaunchUrl
Like (0) or Share (0)