Adaptations of Guest and Host in Expanded Self-Assembled Capsules

Adaptations of guest and host in expanded self-assembled capsules

Dariush Ajami and Julius Rebek, Jr.*

The Skaggs Institute for Chemical Biology and Department of Chemistry, The Scripps Research Institute MB-26, 10550 North Torrey Pines Road, La Jolla, CA 92037

Contributed by Julius Rebek, Jr., August 23, 2007 (sent for review August 2, 2007)

Reversible encapsulation complexes create spaces where two or more molecules can be temporarily isolated. When the mobility of encapsulated molecules is restricted, different arrangements in space are possible, and new forms of isomerism (‘‘social isomerism’’) are created: the orientation of one encapsulated molecule influences that of the other in the confined space. Expansion of a capsule’s length is possible through addition of small-molecule spacer elements. The expanded capsules have dimensions that permit the observation of social isomerism of two identical guests, and they adopt arrangements that properly fill the host’s space. The host also can adapt to longer guests by incorporating additional spacers, much as protein modules are added to a viral capsid in response to larger genomes. Arachidonic and related fatty acid derivatives act in this way to induce the assembly of further extended capsules having sufficient length to accommodate them.

encapsulation ͉ self-assembly ͉ social isomers

ncapsulation complexes are synthetic, self-assembled hosts Ethat more or less completely surround their guest molecules. They are dynamic and form reversibly in solution with lifetimes ranging from milliseconds to days. The capsules isolate molecules from the bulk solution, and they reveal behaviors that cannot be seen otherwise. They have become a tool of modern Fig. 1. The self-assembled cylindrical capule. (Upper) Chemical structure of physical organic chemistry (1). For example, cylindrical host 1.1 the cavitand component and calculated structure of the capsule 1.1 with (Fig. 1) self-assembles only in the presence of suitable guests to peripheral alkyl groups removed. (Lower) Cartoons of the capsule demon- give encapsulation complexes (2–4). Combinations of different strating the social isomers of benzene with p-ethyl toluene. guests (5) inside this capsule have allowed its use as a reaction chamber (6), a chiral receptor (7), and a space where single- molecule solvation can be observed (8). Elsewhere, reversible of the space (16). This matching of host space and guest size encapsulation complexes have been used to stabilize reactive drives much encapsulation and is especially good for arranging intermediates (9, 10) and even transition states (11, 12) as well bimolecular reactions inside (17, 18). It lies at the heart of the as to alter the course of reactions in the limited quarters (13). studies reported here. New forms of stereochemistry also have emerged from coen- Capsule 1.1 reversibly incorporates glycoluril spacers 2 (Fig. 2) capsulation studies (14). For example, with benzene and p-ethyl to give an expanded assembly 1.24.1 with suitable guests (19). toluene, two complexes coexist (Fig. 1). The two molecules are Four glycoluril spacers were added and increased the cavity’s too large to slip past each other, and the p-ethyl toluene is too volume by Ϸ200 Å3 and its length by Ϸ7 Å. A number of longer long to tumble freely while inside the capsule. Either of these alkanes were encapsulated in this host assembly, but the ex- motions could interconvert the complexes, but they are slow on panded capsule also offered broadened possibilities for isomer- the NMR time scale and separate spectra are seen for the two ism and accommodated other guests that we elaborate on here. isomers. These ‘‘social isomers’’ are different arrangements in The results indicate that highly complex molecular assemblies space that arise from the interactions of at least two encapsulated can emerge from only a few modules acting as hosts and guests molecules. They also are constitutional isomers, but mechanical (20, 21). barriers hold these arrays in place rather than covalent bond ‘‘connectedness.’’ Accordingly, the term supramolecular diaste- Results reomers better describes the relationships. This stereoisomerism Simple p-disubstituted benzene derivatives are bound in 1.24.1, is related to the carceroisomerism (15) observed in covalently and the NMR spectra of p-xylene and p-diethyl benzene show bound capsules with one guest. complexes of high symmetry with two guests inside. The spec- The self-assembly of the benzene/p-ethyl toluene pair owes its stability to the optimal filling of the capsule’s space. Two benzene molecules can assemble the capsule as guests in that Author contributions: D.A. performed research; and D.A. and J.R. wrote the paper. solvent and fill 41% of the space. With p-ethyl toluene, one The authors declare no conﬂict of interest. molecule fills only 33% of the space, but two molecules are too *To whom correspondence should be addressed. E-mail: [email protected]. long to be accommodated in the cavity. Accordingly, p-ethyl This article contains supporting information online at www.pnas.org/cgi/content/full/ toluene can be combined with a number of coencapsulation 0707759104/DC1. partners, of which the benzene is at near optimum at filling 53% © 2007 by The National Academy of Sciences of the USA

16000–16003 ͉ PNAS ͉ October 9, 2007 ͉ vol. 104 ͉ no. 41 www.pnas.org͞cgi͞doi͞10.1073͞pnas.0707759104 Downloaded by guest on September 25, 2021 Fig. 2. Formula of the glycoluril 2 spacer and semiempirical energy- minimized structure of the expanded capsule 1.24.1. Both enantiomers and hydrogen bonds are shown, but peripheral alkyl and aryl groups have been removed for viewing clarity. The cartoon schematic used elsewhere in the article is shown on the right.

1 Fig. 4. Upﬁeld region of the H-NMR spectra (600 MHz, mesitylene-d12)of p p-cymene encapsulated in 1.24.1 at various temperatures. The broadening at trum of encapsulated -ethyl toluene, however, shows that three higher temperatures is caused by interconversion of the two enantiomeric species are present (Fig. 3b Left). These are social isomers; they arrays of the glycoluril spacers in the capsules; the social isomers do not reveal that there are several ways to fill the space, but what rules interconvert on the NMR time scale. govern the arrangements? In the most favorable isomer, ethyl groups appear in the narrower center of the capsule, whereas the centrally positioned The chiral nature of the assembly is reflected in the doubling of methyl of the tolyl group penetrates deeper into the tapered the signals of the isopropyl group: its methyls are diastereotopic. At resorcinarene ends. This arrangement apparently creates stron- first glance, the asymmetric elements are far from the cavity’s ger COH/␲ interactions (22, 23) at the ends of the capsule than end, but the chiral arrangement of the glycolurils at the center does a (rotating) ethyl group. Surprisingly, the unsymmetrical of the assembly can shift the eight imide walls in a manner that isomer, although favored statistically, is the highest-energy form is transmitted effectively by the very rigidity of those walls (24). of the three arrangements. The energetic differences between At higher temperatures, the chiral arrangement of the glycolurils social isomers are small (⌬G° varies from Ϸ0.3 to 1.5 kcal/mol) begin to interconvert, and the signals of the diastereotopic and reflect not only the better ‘‘fit’’ in the space but also the methyls of the cymene guest coalesce.

interactions of the coencapsulated molecules. Simple alkanes that are too long to tumble freely in the capsule CHEMISTRY The social isomerism of two p-cymenes provided a surprise: a also show that multiple arrangements are accommodated. Fig. 5 higher stability for the isomer with the methyl toward tapered shows the spectra of 2-methyl heptane and 2-methyl hexane. The ends of the cavitand (as for p-ethyl toluene) was expected, but former shows two of the three possible isomers, whereas the this isomer was not detectable. Instead, the unsymmetrical latter shows only one in the capsule at room temperature. Unlike isomer was favored 2 to 1 over the isomer with both isopropyl alkyl aryls, alkanes with increased rotational degrees of freedom groups near the tapered ends (Fig. 4). Apparently, two isopro- prefer to keep their bulky side in the tapered end of capsule to pyls at the tapered ends or at the constricted middle of the minimize steric clashes in the middle. capsule create steric clashes and do not fit the space comfortably. The seams of hydrogen bonds at the center of the capsular assembly favor polar guests, and this preference was evident in the spectra of N,N-dimethyl-p-toluidine, alkyl anisoles, and aryl alkyl halides. In each case, the major (if not the exclusive) social isomer showed the heteroatoms near the glycolurils [Fig. 6 and supporting information (SI)]. This preference is reinforced by the constricted center of the assembly, which is too narrow to

Fig. 3. Encapsulation of p-dialkylated benzenes. (Left) Downfield and up- 1 field regions of the H-NMR spectra (600 MHz, mesitylene-d12)ofp-xylene (a), 1 p-ethyl toluene (b), and p-diethyl benzene (c) encapsulated in 1.24.1.(Right) Fig. 5. Upfield region of the H-NMR spectra (600 MHz, mesitylene-d12)of The social isomers of p-ethyl toluene related to the spectra in b Left. 2-methyl heptane and 2-methyl hexane in 1.24.1.

Ajami and Rebek PNAS ͉ October 9, 2007 ͉ vol. 104 ͉ no. 41 ͉ 16001 Downloaded by guest on September 25, 2021 Fig. 8. Structures of natural products and derivatives, including arachidonic acid, 4a; anandamide, 4b; oleamide, 5a; an omega-3 ester, 6; and palmitic ethanolamide, 7. All are encapsulated in 1.28.1.

available. The formulation is shown in Fig. 7 as 1.28.1. We probed the adaptability of the larger assembly with a number of natural products having long and narrow shapes and found that they are

Fig. 6. Encapsulation of polar guests. (Upper) Downﬁeld regions of the 1 H-NMR spectra (600 MHz, mesitylene-d12)ofN,N-dimethyl-p-toluidine, 4-methyl anisole, and 1-chloro-3-phenyl propane encapsulated in 1.24.1 and their social isomers in the extended capsule. (Lower) The NMR spectrum and cartoon of encapsulated capsaicin 3 also is shown.

comfortably accommodate substituted aryl groups. The irritant capsaicin 3 isolated from paprika also bears a terminal isopropyl group and is well encapsulated by 1.24.1. The broadened signal for this function suggests an intermediate rate of racemization of the glycoluril array. Preliminary results with normal hydrocarbons longer than C20 showed that an additional belt of glycolurils is incorporated to accommodate these guests when an excess amount of 2 is

Fig. 7. Hyperextension of the cylindrical capsule. (Upper) The energy- Fig. 9. Upﬁeld regions of the NMR spectra of encapsulated arachidonic acid, minimized structure of the doubly expanded capsule 1.28.1. Only one enan- 4a (a); ethyl arachidonate, 4c (b); arachidonyl cyclopropylamide, 4d (c); anan- tiomer is shown, and peripheral alkyl and aryl groups have been removed for damide, 4b (d); methyl docosohexaenoate, 6 (e); palmitoyl ethanolamide, 7 viewing clarity. (Lower) Calculated structure of anandamide 4b in 1.28.1. ( f); 9-octadecenamide, 5a (g); ethyloleate, 5b (h);and 13-docosenamide, 8 (i).

16002 ͉ www.pnas.org͞cgi͞doi͞10.1073͞pnas.0707759104 Ajami and Rebek Downloaded by guest on September 25, 2021 readily encapsulated by the cavitand/glycoluril system (Fig. 8). best interact with the hydrogen bonding seams, and the flexible These include arachidonic acid, 4a, the substrate for the biosyn- guests arrange themselves for a comfortable fit. The hosts do thesis of the prostaglandins, and several of its derivatives, likewise by incorporating additional spacers to provide the including anandamide, 4b, the endogenous ligand for the can- capacity. There are examples in biology that resemble this nabinoid receptor (25). Oleamide, 5a, one of the fatty acid behavior in those viral capsids that add protein units to expand amides involved in sleep induction (26), and its ethyl ester, 5b, the volume inside for longer genomes (28). For applications in also were encapsulated. The lengths of these guests in their the future, we note that two different molecules, each capable of extended conformations reached 26 Å, and they filled between social isomerism in 1.24.1, could result in four different arrange- 40% and 50% of the doubly extended capsule’s space. The ments. All combinations and the chiral nature of the assembly then expand the discrete possibilities to 20, and even many more omega-3 acid methyl ester, 6, showed that at least six cis double are possible if different glycolurils are used simultaneously. If bonds can be accommodated, and the ethanolamide of palmitic these arrangements can be assembled at will, maintained for acid, 7, a saturated fat derivative, also was a good guest. The longer time periods, and read out rapidly, then nanoscale longest guest, at 27 Å, was 8. In the solvent mesitylene-d12, information storage in these capsules would be possible. The slightly more than one equivalent of these molecules is needed encapsulation complexes of the natural products may be useful for complete encapsulation. Fig. 9 (and the SI) shows their NMR in situations where very slow release of biologically active agents, spectra; no evidence of coiling in the alkyl regions took place in one molecule at a time, is desirable. the capsule 1.28.1. The release of these natural products from the capsules can be Materials and Methods accomplished by the addition of hydroxylic solvents. A few Reagents. All reagents were obtained from commercial suppliers percent of methanol added to the mesitylene solvent disrupts the and used without further purification. NMR spectra were re- hydrogen bonds and destroys the assembly. The weakly basic corded on a Bruker DRX-600 spectrometer. The NMR samples sites of the glycolurils offer a site for protonation, and when acids were prepared as follows. For capsule 1.1, a 2 mM concentration are added to the mesitylene solvent, precipitation of the pro- of 1 and 1–20 mM concentrations of the appropriate guest were tonated glycolurils occurs with release of the guests. The assem- mixed with 0.6 ml of mesitylene-d12 in a NMR tube. The tube was bly can be restored by the subsequent addition of tertiary amines placed in an ultrasonic bath (230 W) and sonicated for 5–10 min. (27). Accordingly, the uptake and release of these guests can be Capsules 1.24.1 and 1.28.1 were prepared similarly to the above controlled through the polarity of the medium or acid/base procedure, except for the addition of glycoluril 2 (4 mM for 1.24.1 chemistry. and 8 mM for 1.28.1) to the mixture. Discussion 1H-NMR Spectra. Complete 1H-NMR spectra of encapsulated The multiple host/guest arrays shown by these assemblies differ N,N-dimethyl-p-toluidine, aryl alkyl halides, aryl alkyls, and natural products 3-8 are available in the SI. from a ‘‘lock and key’’ recognition paradigm; there are several energetically comparable ways to fill the space given a specified We thank Prof. B. Cravatt (The Scripps Research Institute) for samples ‘‘filling.’’ Rather, the term ‘‘induced fit’’ is more apt if it applies of the fatty acid derivatives. We are grateful to The Skaggs Institute and to the behavior of both partners; they mutually adapt for the the National Institutes of Health (Grant GM 50174) for financial CHEMISTRY optimal filling of space. The polar guests position themselves to support. D.A. is a Skaggs Postdoctoral Fellow.

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