733

Applications
Yoonkyung

of dendrimers

in bio-organic

chemistry

Kim and Steven

C Zimmerman*
This review will broadly focus on applications of dendrimers in bio-organic chemistry that have been reported in the past year; and, although somewhat different in focus, it updates earlier reviews published [3 ,4]. An excellent review of dendrimers that act as biological mimics was recently published by Smith and Diederich [S Beyond ]. discussing a couple of results in the area of biomimetic dendrimers, this paper will highlight work on dendrimers whose monomer units are derived from carbohydrates, amino acids or nucleotides, and also several promising biomedical applications under active investigation. For readers interested in a historical account, terminology, nomenclature, or other background material, an excellent monograph by Newkome etal. [l] is recommended.

Dendrimers represent a new class of highly branched polymers whose interior cavities and multiple peripheral groups facilitate potential applications in biomedicine and bio-organic chemistry. Major advances in the past year were made in the synthesis and study of new carbohydrate, nucleic acid, and peptide dendrimers, as well as in the use of dendrimers as magnetic resonance imaging contrast agents, as agents for cellular delivery of nucleic acids, and as scaffolds for biomimetic systems.

Addresses Department of Chemistry, 600 S Mathews Avenue, University of Illinois, Urbana, IL 61801, USA
*e-mail: sczimmer@uiuc.edu Current Opinion in Chemical Biology 1998, 2:733-742

http://biomednet.com/elecref/1367593100200733 0 Current Biology Ltd ISSN 1367-5931

Synthesis of dendrimers
Principle efforts in dendrimer chemistry over the past decade have focused on developing new synthetic strategies and on structural variation. With a remarkably diverse set of dendrimers available, it is logical and desirable that attention shift to physical studies and applications. Nonetheless, it remains imperative that more efficient and scalable syntheses be developed for physical property measurements and commercial applications to proceed at a reasonable pace. Currently, only the divergent synthetic approach [l] is capable of producing kilogram quantities of higher generation dendrimers, and only two classes of dendrimers (PAMAM and poly[propyleneimine], PPI) are commercially available. On the other hand, the convergent approach produces dendrimers with fewer defects. In normal convergent dendrimer syntheses the A group in the AB, monomer must be protected or require activation to avoid premature reaction with the B group (the roles of A and B are reversed in divergent syntheses). An improvement on Frechet s important branched monomer (convergent) approach to dendrimer synthesis [6] was recently described [7]. This approach, known as the orthogonal coupling strategy, uses two orthogonal monomers (AB, and A ,, where A reacts only with B and B A only with B) to obviate the need for the deprotection or activation steps, and thus represents the shortest synthetic approach in terms of steps. Whereas the AB, monomer (n = 2, 4) used in this approach [7] produced layered (i.e. block copolymer) dendrimers, Yu and co-workers [8 have ] shown that the same orthogonal strategy can also produce homogeneous poly(phenylene-vinylene) dendrimers. As outlined in Figure 2, this new dendrimer synthesis uses the Horner-Wadsworth-Emmons reaction of AB, monomer 1 in alternation with a Heck reaction using the second AB, monomer, 2. Equally critical to the need for short syntheses is the need for dendrimers with a small number of reactive functional groups at the surface or interior for subsequent, controlled

Abbreviations Boc ferf-butoxycarbonyl DTPA MAP NCA PAMAM PPI diethylenetriaminepentaacetic multiple antigen peptide N-carboxyanhydride poly(amidoamine) poly(propyleneimine) acid

Introduction
Dendrimers are a relatively new class of polymers with structures that depart rather dramatically from traditional linear polymers, which are built from so called AB monomers [l]. Because they are built from AB, monomers (where n 2 2, and is usually 2 or 3), dendrimers are highly branched and have three distinct structural features: a core, multiple peripheral (end-)groups, and branching units that link the two. The peripheral groups and branching units are together called dendrons, or informally, wedges . Branching units used to date have contained virtually every type of functional group ranging from those comprising pure hydrocarbons or aromatic groups to modified carbohydrates and nucleic acids. Dendrimers are synthesized iteratively so that the number of intervening branching units between the core and one end-group (i.e. the number of layers), called the generation number, is determined by the number of synthetic cycles. In the commercially available poly(amidoamine) (PAMAM) dendrimers, ammonia represents the zeroth generation (see Figure 1) [Z]. As the generation number increases from one to seven, the number of peripheral groups follows the geometric series 3, 6, 12 . . . 192. These peripheral groups largely control the solubility of the compound, so that even with highly hydrophobic internal units a dendrimer will dissolve in water if the end groups are sufficiently hydrophilic. As will be discussed here, this feature and the high local concentration of end groups enables a number of applications in bio-organic chemistry.

734

Model systems

Figure

sutface/end-groups

Currrent Opinion in Chemical

Biology

A generational series

of commercially available PAMAM dendrimers core. (a) Triple Michael addition with

chain extension

gives (c) the second-generation

dendrimer, and

with an ammonia-derived

then (d) the third-generation

dendrimer. The other major class of

methylacrylate followed by chain extension with ethylene diamine takes the zeroth generation (core) to (b) the first-generation dendrimer. Synthesis is iterative, so repeating Michael addition and

commercially available dendrimers are represented by (e) the thirdgeneration Meijer PPI dendrimer, which is derived from 1,4_butanediamine and acrylonitrile.

modification (e.g. attachment of ligands for cell surface binding). The PAMAM dendrimers have been used as a scaffold to hold two different proteins, for example. Singh [9] sequentially reacted a fifth-generation PAMAM dendrimer with N-hydroxysuccinimidyl iodoacetate, calf intestine alkaline phosphatase, and a Fab fragment (the antigen-binding fragment of an antibody) of an anti-creatine kinase MB isoenzyme antibody to create a multifunctional biochemical reagent. The number of proteins attached can be controlled to a degree but the relative positioning cannot. For example, in a third-generation PAMAM dendrimer (Figure Id), modification of one amino group on the surface leaves three distinct types of amino groups so the second modification can make three different compounds. The problem will be exacerbated for higher generation dendrimers, which have many more surface groups. For many applications, the reported level of control may be adequate, but clearly for many others,

especially those with a precise number of clustered a more advanced dendrimer will be needed.

groups,

The construction of dendrimer libraries will represent a very important new direction for optimization of desired properties, or for finding lead structures. At the time of writing, combinatorial libraries of discretely synthesized macromolecules were unknown. Dendrimers are ideal scaffolds for combinatorial chemistry because their synthesis is iterative and the monomeric building blocks are simple, so a structurally diverse set of AB, monomers can be readily prepared. The structural variation may be between layers, wedges, or within one or more layer. For example, Newkome et al. [lo] reacted a fourth-generation PPI dendrimer (which has 32 amino end-groups) with a mixture of two AB, isocyanates. Because the isocyanates were of comparable reactivity to one another, the newly added outer layer of tert-butoxycarbonyl

Applications

of dendrimers

in bio-organic

chemistry

Kim and Zimmerman

735

Figure

Br

Br

1:
Em-A pa
Ed 0 .OEt bP OEt

1:
Em-A pa
Ed 0 .OEt bP OEt

Pd(OAc)s Bu4NBr, K&O3 56%

Current Opinion in Chemical Biology

An extension

of the Zeng and Zimmerman s

[3 orthogonal ]

coupling

The second

orthogonal

reaction

involves palladium-catalyzed

Heck

approach to dendrimer synthesis that produces structurally homogeneous dendrimers. The synthesis of the poly(phenylene-vinylene) dendron bromide 6 proceeds from 3+4+5+6 without any deprotection or activation steps. One orthogonal reaction, the sodium hydride mediated 1 and an (NMP). Horner-Wadsworth-Emmons reaction of his-phosphonate aromatic aldehyde (e.g. 3) occurs in N-methylpyrrolidinone

coupling of dialkene 2 and an aryl aldehyde (e.g. 4). The disadvantage of this approach is the need for two different AB, monomers (in this case, 1 and 2, where n = 2) and two orthogonal coupling reactions; however, this strategy allows one generation to be added with each iteration, thereby with standard doubling the rate of dendrimer approaches. AC, acetyl; growth when compared Bu, butyl; Et, ethyl.

(Boc)-protected amino groups contained a mixture of groups derived from both isocyanates. These end-groups were subsequently deprotected and reacted with a mixture of three isocyanates to give a new, mixed layer of end-groups [lo]. The number of isomers produced in such a scheme is astronomical, so this intriguing chemistry is most likely to be used to create a single compound with a broad range of properties, or to compare the properties of compounds prepared from different reagent stoichiometries. Some effort has been invested in modifying groups on the interior of dendrimers. In fact, Caminade, Majoral and coworkers [ll] have shown that selective activation of internal functionality can lead to dendrimeric growth within the confines of a dendrimer. A third-generation dendrimer can be selectively activated at six internal phosphorous atoms, allowing iterative growth of internal dendrimers of the second to fourth generation (7, a fourth-

generation dendrimer; Figure 3). These results not only emphasize the considerable flexibility and void size of small dendrimers, but they also point the way to synthesizing novel hybrid dendrimeric structures with potential applications in complexation and catalysis.

Carbohydrate

dendrimers

The groups of Lindhorst, Okada, Roy, and Stoddart were instrumental in developing the chemistry of carbohydrate dendrimers, an area that has been admirably reviewed by Stoddart and co-workers [12]. Since the publication of this review, significant advances that have been reported include the chemoenzymatic synthesis of dendrimers with peripheral N-acetyl lactosamine units [13]. As shown in Figure 4, a chemically synthesized third-generation lysine dendrimer with unprotected peripheral N-acetyl glucosamine units (8) was enzymatically converted to N-acetyl lactosamine 9, in good yield. This particular disaccharide plays a role in disease states

736

Model systems

Figure 3

7
A third-generation dendrimer can be selectively activated at six internal bold arcs. The chemical structure

Current Opinion in Chemical

Biology

suggests

that this new growth end-groups are indicated by

phosphorous atoms (see bold bonds). These six positions become the core for the divergent growth of a second set of dendrimers. The new dendrimeric growth terminates in benzaldehyde units, indicated by the

actually engulfs the first dendrimer, whose the arrows. Me, methyl; Ph, phenyl.

(e.g. mouse colon cancer, thyroid disorders), but the broader importance of the work is that highly regioselective and stereoselective enzymatic saccharide couplings can obviate the chemical couplings, which are rarely 100% selective. A small isomeric impurity in

an early chemical coupling will be magnified as the dendrimer generation increases. For example, a 1% impurity (or defect) in an AB, monomer means that at the fifth generation over 25% of the compound will contain the defect.

Applications

of dendrimers

in bio-organic

chemistry

Kim and Zimmerman

737

Figure

H H&sJ~,y-~,y NHAc

H H&SdG,y_;,y/j NHAc

UDP-Glc GlcNac p-1,4-Gal-transferase UDP-Glc 4 -epimerase calf intestinal phosphatase

HOH&sdG,y_G,y NHAc \

/y=p-A a-oH

HOH&sdG,,.,j
NHAc \i H H&S&y-G,y NHAc i HOH&slG,y_G,) NHAc

9
Current ODinion in Chemical Bioloav

Zanini and Roy [13] chemoenzymatic conversion of a s monosaccharide to a disaccharide on the surface of a polylysine dendrimer. The reaction from 8 (a chemically synthesized, thirdgeneration lysine dendrimer, with unprotected peripheral N-acetyl gluocosamine [GlcNAc] units) to 9 (with peripheral N-acetyl

lactosamine units) proceeds in high yield and presumably with near perfect regioselectivity and stereoselectivity. The ability to carry out an enzymatic process on the surface of a dendrimer will obviate the need for involved protection and deprotection chemistry. AC, acetyl; Gal, galactose; Glc, glucose; IJDP, uridine monophosphate.

Okada and co-workers [14] has continued to develop the chemistry of these so-called sugar balls, and reported an interesting radial-growth polymerization . This process involves the 24,48, or 96 terminal amino groups of a third-, fourth- or fifth-generation PAMAM dendrimer, each initiating a ring-opening polymerization of a protected 0-glucosyl-L-serine-N-carboxyanhydride (NCA). These polymerizations were carried out at room temperature for 24-80 hours to give star-like block co-polymers (layers) with low polydispersity values and low degrees of polymerization (DP = 3-4). (In general, low polydispersities are advantageous in allowing one to work with material that has a narrower range of properties.) One of the most frequently mentioned applications of carbohydrate dendrimers is to increase the modest receptor binding affinities of monosaccharides through a multivalent interaction (known as the cluster effect ). Cell surfaces present one obvious target for such agents, and the biomedical implications are obvious. The success of this approach will require a balance between the degree of preorganization, water solubility, local concentration of the saccharide, and its accessibility. With these issues in mind, Meijer, Stoddart and co-workers [15] reported the synthesis of a PPI dendrimer with as many as 64 surface glucosyl units, but with linker arms of either one, five or ten methylene groups intervening between the sugar and the

dendrimer. This idea that multivalency will afford higher bioactivity for carbohydrate dendrimers relative to the analogous monosaccharides has received support: Zanini and Roy [16] reported that an a-thiosialosyl-coated fourthgeneration PAMAM dendrimer inhibited the binding of human a, acid glycoprotein to Limax flaws (slug lectin) at a concentration of 1 nM, representing a ZlO-fold increase over a mono-thiosialoside; a tetravalent, first-generation galabioside inhibited bacterial adhesion at 2 nM concentration, which is a 600-fold improvement over a monomeric galabioside derivative [17]. Other applications may stem from the ability of dendrimers constructed entirely of carbohydrates to bind small molecules on their interiors. Thus, these dendrimers may form cavities on their interiors, which are bounded by the hydrophobic surface of the sugar units in much the same way that cyclodextrins include hydrophobic molecules. In this vein, Stoddart and co-workers [18 has reported sig] nificant strides toward the synthesis of such dendrimers.

Peptide dendrimers
Many dendrimers contain amide linkages (e.g. the PAMAM class), but few are built from naturally occurring amino acids. Interestingly, one of the earliest reports of a dendrimer, a patent by Denkewalter et al. [Pl], reported the synthesis of lysine dendrimers up to the

738

Model systems

tenth generation. To convert other amino acids from ABtype to AB,-type monomers (lysine is already AB,), either the carboxylic acid group or the amino group must be derivatized to create a branch point. Liskamp and co-workers [19] have accomplished this by etherification of methyl 3,5-dihydroxybenzoate with Boc phenyl-alaninol. This monomer was iteratively converted to a third-generation dendrimer, and the authors noted their intention of using other amino acid monomers to create a combinatorial library of peptide dendrimers. Tam has pioneered the synthesis and study of a different type of peptide dendrimer (see [ZO] for lead references). These multiple antigen peptides (MAPS) contain a lysine dendrimer scaffold holding multiple copies of an antigenic peptide, which enhance its immunogenicity. All peptide dendrimers have potential applications that include their use as artificial enzymes, drug delivery vehicles, and pharmaceuticals. This is clearly an area ripe for development and it will be particularly interesting to see if peptide dendrimers based on natural amino acids can be engineered to fold into specific structures.

Dendrimers as vehicles for delivery of nucleic acids

The difficulty in getting nucleic acids into cells has hampered efforts to develop both antisense therapeutics and gene therapies. Considerable attention has focused on nonviral gene/antisense delivery methods using cationic liposomes and polymers such as polylysine and PAMAM dendrimers. Early studies were promising, showing that PAMAM dendrimers are significantly more efficient and less toxic than polylysine and that they stabilize oligonucleotides within the cell [3 ,26]. Thus, not surprisingly, considerable recent effort has sought to elucidate the mechanism by which dendrimers effect these improvements. DeLong et al. [27] showed that a third-generation PAMAM dendrimer forms stable complexes with phosphorothioatelinked oligonucleotides (which are useful in the antisense approach) with molecular weights that are mostly in the intermediate lo-100 kDa range. The 1:l and 2O:l complexes (dendrimer : oligonucleotide) increased the cellular uptake by three to fourfold and SO-fold, respectively. These results contrast those of Tomalia, Baker and co-workers [28] who found that efficient complexation and facilitated cellular uptake of DNA occurred only when higher generation (2 5) PAMAM dendrimers were used.

Dendritic nucleic acids

Two types of dendrimeric nucleic acids have been studied: those that are covalently connected and those that selfassemble via complementary base-pairing schemes. A theoretical study of those that self-assemble by complementary base-pairing was reported, wherein it was shown that unique oligonucleotide monomers could hybridize complementary single strands so that a sixth-generation dendrimer would be self-assembled from 1,457 monomer units [Zl]. An example of a covalent DNA dendrimer was reported by Shchepinov et al. [ZZ], wherein a second-generation phosphodiester-based dendrimer derived from pentaerythritol held nine pentathymidylate units on the end-groups and either a 15mer or 26-mer at the core. The authors showed that the core sequence hybridizes a complementary strand linked to a solid support, with the dendrimer portion giving an enhanced phosphorimage when phosphorylated with 32P. The core sequence was also competent as a PCR primer, and therefore could easily be used to amplify the dendritic DNA. The authors suggest that these types of dendrimeric nucleic acids may be useful DNA probes with advanced signaling potential and higher labeling capacity. In fact, Chiron Quantiplex assay s system (used to quantify viral DNA or RNA in blood or serum) uses branched DNA with a comb-like structure to hybridize target DNA sequences, and subsequently amplify the reporter signal [23,24]. Such amplification has been necessary for clinical and diagnostic applications because PCR techniques cannot reproducibly quantify the amount of DNA in a sample. Hudson etal. [ZS] have developed the chemistry to make dendrimers divergently and entirely of nucleic acids. In their systems, DNA oligomers serve as spacers to link the branch points, which are 2 and 3 -linked ribonucleotides.

Dendrimers as magnetic resonance imaging contrast agents

In a recent review [3 it was noted that Wiener etal. [29] ], developed dendrimeric polygadolinium chelates as improved magnetic resonance imaging contrast agents. The agents, prepared by the reaction of PAMAM dendrimers and gadolinium-chelated Z-(4-isothiocyanato-benzyl)-6methyl-diethylenetriaminepentaacetic acid (DTPA), showed enhanced proton relaxation when compared with other polymeric or monovalent chelators. Wiener et al. [30] improved the targeting of this early system to certain cancer cells by conjugating folic acid to a fourth-generation PAMAM dendrimer followed by linking to DTPA. A highaffinity folate receptor is expressed or upregulated on the surface of several types of cancers, including those of the ovary, endometrium and breast. In such cells, a 110% increase in longitudinal relaxation rate was directly attributed to folate recognition, indicating the utility of this approach in cancer treatment. Other recent advances include the conjugation of PAMAM dendrimers to a macrocyclic ligand, tetraazacyclododecane-4,7,10_triacetic acid, which forms complexes with Gd3+ that are kinetically more stable than those formed from DTPA [31]. To reduce the liver uptake and increase bioavailability, polyethylene glyco1 (PEG) chains were grafted onto the residual amino groups on the PAMAM periphery. (Liposomes and polymeric drug delivery systems coated with PEG sidechains are less likely to be carried to the liver through the reticuloendothelial system.)

Biomimetic dendrimers
One of the most active areas of dendrimer research involves their use in biomimetic systems. Dendrimers

Applications

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in bio-organic

chemistry

Kim and Zimmerman

739

with metalloporphyrin cores have been especially popular with major efforts reported by Aida and co-workers [32 ,33], Diederich and co-workers [34,35-l, Frechet and co-workers [36,37] and Moore, &slick and co-workers [38]. Differences between studies in these laboratories can be found in the type of dendrimer used and the synthetic approach. Diederich and co-workers used the divergent approach to synthesize 10 (Figure 5) and other lower-generation analogs holding Newkome-type [39] poly(ether-amide) dendrons. In contrast, the other investigators have used the convergent approach, which can limit the size (and yield) of the dendrimer produced, but generally leads to purer materials. Aida and co-workers [32 ,33] have attached first through fifth-generation Frechet-type dendrons to a porphyrin core (e.g. 11, M = Fe, R = CH,), as have Frechet and co-workers [36] (11, M = Zn, R = benzyl, up to fourth generation). Often, too little effort is invested in exploring the synthetic approaches to these sort of compounds, so it was encouraging that Frechet and co-workers [36] compared two different convergent approaches to the synthesis of 11. The first is similar to the approach used by Aida, Moore, &slick and their co-workers [32 ,33,38], in which the dendrons are attached to a preformed tetrakis(3,5dihywhereas the other involves droxyphenyl)porphyrin,
Figure 5

condensation the Lindsey

of pyrrole and a dendrimeric porphyrin synthesis.

aldehyde

via

Several investigators have examined how the globular dendrimeric shell in these models affects the redox potential of the metalloporphyrin [34]. The results of these studies from two different laboratories have been discussed recently by Frechet and co-workers [37] and will not be covered here. Other structures of bio-inorganic interest have been placed within a dendrimeric shell, most notably an electroactive iron-sulfur core, (Fe&&[SR],)2(where R = dendrimeric substituent), studied by Gorman et al. [40]. As the dendrimeric arms increase in size from the first to fourth generation, the reduction potential becomes more negative. The larger dendrons are believed to act as insulators. Another intriguing avenue of research has involved the use of dendrimeric porphyrins as reversible oxygen binders (i.e. hemoglobin mimics). Jiang and Aida [32 have shown that ] the fifth-generation analog of 11 (Figure 5) in water-saturated toluene, with l-methylimidazole, reversibly binds dioxygen and forms an 0, complex that is stable for more than 2 months. Collman, Diederich and co-workers [35 ] studied a first- and second-generation analog of 10 (M = Fe, R = CO[OCH,CH,],OMe, where Me is methyl) and found

Current Opinion in Chemical Biology

Several groups have examined the electrochemical properties, catalytic function, and oxygen-carrying ability of dendrimeric porphyrins. Compound 10 (R = CO&H,) and lower generation analogs were [34,35-l, while prepared and studied by Diederich and co-workers

dendrimeric porphyrin 11 and generational analogs were prepared and studied by Aida and co-workers [32 ,33] (M = Fe; R = CH,) and Frbchet and co-workers [36,37] (M = Zn; R = benzyl).

740

Model systems

extremely high-affinity binding of 0, in toluene with 1,2methylimidazole as an axial ligand. The 0, affinity was about 1,500-fold higher than that of hemoglobin (in its tense [T] state, with a lower affinity for oxygen), and, combined with the weaker binding of CO, gave a low M value (PI/~ [O,]/ Ps [CO]) of 0.08 (where Ps is partial pressure for half occupancy). The origin of this tight oxygen binding may result from the formation of a hydrogen bond from oxygen to the dendrimeric shell. The use of these compounds for biomimetic catalysis will certainly be explored. For oxidative chemistry, unreactive dendrimeric groups will be needed to prevent catalyst self-oxidation. Moore, &slick and co-workers [38] used poly(phenylester) dendrons that were inert to their shape-selective epoxidation reaction. Some effort has been focused on making dendrimers in which the monomer units are porphyrins. A first-generation system with five porphyrins was synthesized by Norsten and Branda [41] and is related to an earlier structure reported by Officer et al. [42]. Van Veggel, Reinhoudt and co-workers [43 have used their imagi] native palladium coordination chemistry with branched monomers to assemble a second-generation metallodendrimer with as many as 12 porphyrin units on the surface. This is an elegant approach that minimizes the number of synthetic steps. An exciting and important area that has received considerable attention, yet remains in an embryonic state, is the use of dendrimers as synthetic hosts for small molecules. Earlier reviews [3 ,4] highlighted Meijer demonstration s [44] that small molecules (e.g. Bengal rose) could be mechanically trapped within a PPI dendrimer, and then released upon chemical cleavage of surface functionality [44]. This system was examined computationally by Goddard and co-workers [45], with remarkable agreement found between experiment and theory. Cavallo and Fraternali [46] examined the PPI dendrimers computationally without the guest (i.e. ligand) molecules present, and found, for the higher generations, a denser packed structure with back-folded end-groups that do not totally fill the voids. It is argued that such a structure could entrap molecules. The fact that dendrimers function as hosts (receptors) to nonspecifically complex guest molecules within their interiors is well established. A recent report by Spinder, Tomalia and co-workers [47] described the reaction of aliphatic epoxides with PAMAM dendrimers, converting them into inverse micelles (for copper transport), similar to Meijer and co-workers [48] recent inverted micelle work using PPI dendrimers. An important finding, with obvious and broad ranging applications (such as removal of organic pollutants from waste water) is that PPI dendrimers that are surface-modified with fluorocarbon chains can dissolve in liquid carbon dioxide and extract polar organic molecules from water [49 Examples of ]. multisite recognition involve dendrimers with multiple

ferrocene units as end-groups. Kaifer and co-workers [SO] found that such dendrimers can simultaneously bind multiple cyclodextrins, while Astruc and co-workers [Sl] showed their potential use as selective redox sensors for common inorganic anions. With regard to specific recognition, it was shown that dendrimeric naphthyridines will bind amidinium ions at their core in organic solvents [SZ]. Remarkably, with two fourth-generation dendrons of either the Moore-type (rigid, apolar diphenylacetylene) or the Frechet-type (flexible, polar phenylbenzylether) at the 2,7-positions of naphthyridine, there was no influence on the complexation strength. Furthermore, increases in solvent polarity causes a decrease in binding strengths. The combined results suggest that the hosts are porous and filled with solvent. The observation that hydrogenbonding interactions can occur at dendrimer cores suggests the exciting possibility of using hydrophobic dendrons with water-solubilizing end-groups so that hydrogen-bond complexation might occur in water by virtue of the micellar effect (hydrophilic exterior, hydrophobic interior). If this is possible, the many synthetic hosts that bind their guests by hydrogen bonding in apolar organic solvents, might be derivatized so that they function well in an aqueous medium. In this regard, Diederich and co-workers [53] have shown specific binding of simple aromatic compounds and steroids by a dendrophane , a host built by attaching Newkome-type dendrons to a well studied cyclophanic host.

Conclusions
Progress over the past year clearly demonstrates that dendrimers are no longer just novel and aesthetically appealing polymers. Their unique structural features and properties make them ideally suited for both a wide range of biomedical applications and as platforms for biomimetic chemistry. Further advances in synthetic approaches will only hasten the rate at which practical uses for dendrimers are found. It is important to note that hyperbranched polymers, prepared by polymerization of AB, monomers, may offer a cheaper alternative to dendrimers. Methods for producing these less perfect macromolecules with lower polydispersities may advance to the level where their physical properties are nearly indistinguishable from their perfect counterparts, dendrimers.

Acknowledgement
This work was funded by the National Institutes of Health.

References and recommended

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l

reading
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the annual

of special

interest interest Moorefield CN, VGgtle F: Dendritic Weinheim: Molecules: VCH; 1996.

**of 1.

outstanding GR,

Newkome Concepts,

Syntheses, DA,

Perspectives.

2.

Tomalia

Naylor AM,

molecular-level

Goddard WA III: Starburst dendrimers: control of size, shape, surface chemistry,

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topology, and flexibility from atoms to macroscopic Chem ht Ed Engll990, 29:138-l 75. 3. .

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Nilesen TW, Grayzel J, Prensky W: Dendritic nucleic acid structures. Biol 1997, 1871273.284.

Zeng F, Zimmerman SC: Dendrimers in supramolecular chemistry: from molecular recognition to self-assembly. Chem Rev 1997, 97:1681-l 712. A comprehensive review of molecular recognition and self-assembly studies using dendrimers and their implications for applications in biomedicine and as advanced materials. 4. Zimmerman SC: Dendrimers assembly. Curr Opin Colloid in molecular recognition and selfSci 1997, 2:89-99.

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Shchepinov MS, Udalova IA, Bridgman AJ, Southern EM: Oligonucleotide dendrimers: synthesis and use as polylabelled DNA probes. Nucleic Acids Res 1997, 25:4447-4454. Urdea MS: Synthesis and characterization of branched DNA (bDNA) for the direct and quantitative detection of CMV, HBV, HCV, and HIV. C/in Cbem 1993, 39:725-726. Urdea MS: Branched DNA signal amplification. 1994, 12:926-927.
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Smith DK, Diederich FN: Functional dendrimers: unique biological mimics. Chemistry - Eur J 1998,4:1353-l 361. This is an excellent review of selected biomimetic systems based on dendrimers. 6. Wooley KL, Hawker CJ, Frechet JMJ: A branched-monomer approach for the rapid synthesis of dendrimers. Angew Chem Ed Engll994, 33:82-85. Zeng F, Zimmerman SC: Rapid synthesis of dendrimers orthogonal coupling strategy. J Am Cbem Sot 1996, 11815326.5327. by an

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Hudson RHE, Robidoux S, Damha M: Divergent solid-phase synthesis of nucleic acid dendrimers. Tetrahedron Lett 1998, 39:1299-l 302. Haensler J, Szoka FC Jr: Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjug Cbem 1993,4:372-379. DeLong R, Stephenson K, Loftus T, Fisher M, Alahari S, Nolting A, Juliano RL: Characterization of complexes of oligonucleotides with polyamidoamine starburst dendrimers and effects on intracellular delivery. J fbarmacol Sci 1997, 86:762-764. Kukowska-Latallo JF, Bielinska AU, Johnson J, Spindler R, Tomalia DA, Baker JR Jr: Efficient transfer of genetic material into mammalian cells using starbust polyamidoamine dendrimers. froc NatlAcad SC; USA 1996, 93:4897-4902. Wiener EC, Brechbiel MW, Brothers H, Magin RL, Gansow OA, Tomalia DA, Lauterbur PC: Dendrimer-based metal chelates: a new class of magnetic resonance imaging contrast agents. n/lagn Reson Med 1994,31 :l -8. Wiener EC, Konda S, Shadron A, Brechbiel M, Gansow 0: Targeting dendrimer-chelates to tumors and tumor cells expressing the high-affinity folate receptor. /west Radio/ 1997, 32:748-754. Margerum LD, Campion BK, Koo M, Shargill N, Lai J-J, Marumoto A, Sontum PC: Gadolinium(lll) D03A macrocycles and polyethylene glycol coupled to dendrimers. Effect of molecular weight on physical and biological properties of macromolecular magnetic resonance imaging contrast agents. J A//oys Compounds 1997, 249:185-l 90.

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