Aggregation Behavior of Nucleoside-Boron Cluster Conjugates in

Aqueous Solutions
Pavel Matějı́ček,† Petr Cı́gler,‡,§ Agnieszka B. Olejniczak,| Agnieszka Andrysiak,|
Blazej Wojtczak,| Karel Procházka,† and Zbigniew J. Lesnikowski*,|
Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles UniVersity in Prague,
HlaVoVa 2030, 128 43 Prague 2, Czech Republic; Gilead Sciences and IOCB Research Center, Institute of
Organic Chemistry and Biochemistry, AS CR, FlemingoVo n. 2, 166 10 Prague 6, Czech Republic;
Department of Analytical Chemistry, Institute of Chemical Technology, Prague, Czech Republic; and
Laboratory of Molecular Virology and Biological Chemistry, Center of Medical Biology,
Polish Academy of Sciences, 106 Lodowa St., Lodz 93-232, Poland

ReceiVed September 13, 2007. In Final Form: NoVember 14, 2007

We report the first evidence that boron-containing nucleoside conjugates have a tendency to associate in water
solutions. The size, charge, and exoskeletal pattern of the boron cluster can strongly influence the aggregation. The
aggregation of nucleosides with attached boron clusters was observed using light scattering and atomic force microscopy
techniques. Although the species containing either the bulky amphiphilic [3-cobalt(III) bis(1,2-dicarbollide)]- anion
or the electroneutral dicarba-closo-dodecaboranyl moiety tend to form stable nanoparticles in aqueous solutions, the
compounds bearing the smaller, negatively charged dicarba-nido-undecaboranyl moiety as well as the unmodified
nucleosides do not aggregate. The light scattering measurements also showed that the aggregated species can interact
with nonionic surfactant Triton X-100 in solution. The partition coefficients P in the water-octanol system correlate
fairly well with the aggregation tendency observed by light scattering measurements. This finding allows us to predict
the association behavior of boron-cluster-containing nucleosides on a qualitative level. The observed phenomenon
can contribute to a better understanding of biological properties of boronated nucleosides and the design of boronated
nucleoside-based drugs such as boron carriers for boron neutron capture therapy of tumors (BNCT) and antiviral
agents.

Introduction construction of novel pharmaceuticals and biological probes.5,10

The original stimulus for the design and synthesis of boron-
containing nucleosides was their potential application as rich
boron carriers for boron neutron capture therapy (BNCT).1-4
More recently, it has been demonstrated that carboranes enhance
hydrophobic interactions between boron-coupled pharmaceuticals
and their receptors and increase the in vivo stability of compounds
that otherwise would be rapidly metabolized.5-8 The carborane
modification also facilitates cellular uptake because of the
increased lipophilicity and easier penetration of the carborane-
bearing compounds through cellular membranes.9 These proper-
ties, together with a diverse chemistry of boron clusters, make
the boron-containing nucleosides attractive synthons for the
* To whom correspondence should be addressed. E-mail:
zlesnikowski@cbm.pan.pl. Tel: 048-42-2723629. Fax: 048-42-2723630.
† Charles University in Prague.
‡ Institute of Organic Chemistry and Biochemistry.
§ Institute of Chemical Technology.
| Polish Academy of Sciences.
(1) Sjöberg, S.; Carlsson, J.; Ghaneolhosseini, H.; Gedda, L.; Hartman, T.;
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(5) Yamamoto, K.; Endo, Y. Bioorg. Med. Chem. Lett. 2001, 11, 2389.
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Chem. 2006, 17, 928.
Indeed, several antiviral nucleosides have already been modified
with carborane cages, and their biological properties have been
studied.2 Some carborane clusters have also been explored as
new and versatile modifying entities for DNA oligonucle-
otides.11,12 From this perspective, carborane-containing nucleo-
sides are desirable precursors for the preparation of modified
DNA oligomers. In addition, carboranes were used as the cores
from which a series of potent estrogen6 and retinoid7 agonists
or transthyretin dissociation inhibitors13 were constructed.
Another specific group of carborane derivatives is represented
by metallacarboranes. After a long period of neglecting the study
of their biological properties, a number of research groups have
recently started to investigate their anti-viral and anti-tumor
activities14-16 and their potential use as boron carriers in
BNCT.9,17-20 A special advantage of metallacarborane nucleo-
(10) Valliant, J. F.; Guenther, K. J.; King, A. S.; Morel, P.; Schaffer, P.;
Sogbein, O. O.; Stephenson, K. A. Chem. ReV. 2000, 232, 173.
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Pokorna, J.; Plesek, J.; Gruner, B.; Doleckova-Maresova, L.; Masa, M.; Sedlacek,
J.; Bodem, J.; Krausslich, H. G.; Kral, V.; Konvalinka, J. Proc. Natl. Acad. Sci.
U.S.A. 2005, 102, 15394.
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Grimes, R. N. Anticancer Res. 2000, 20, 2345.
(16) Kubát, P.; Lang, K.; Cı́gler, P.; Kožı́šek, M.; Matějı́ček, P.; Janda, P.;
Zelinger, Z.; Procházka, K.; Král, V. J. Phys. Chem. B 2007, 111, 4539.
(17) Lesnikowski, Z. J.; Paradowska, E.; Olejniczak, A. B.; Studzinska, M.;
Seekamp, P.; Schussler, U.; Gabel, D.; Schinazi, R. F.; Plešek, J. Bioorg. Med.
Chem. 2005, 13, 4168.
(18) Hao, E.; Vicente, M. G. Chem. Commun. 2005, 10, 1306.
(19) Hao, E.; Jensen, T. J.; Courtney, B. H.; Vicente, M. G. H. Bioconjugate
Chem. 2005, 16, 1495.

10.1021/la702852e CCC: $40.75 © xxxx American Chemical Society

Published on Web 02/08/2008 PAGE EST: 5.3
B Langmuir
sides is their application in the synthesis of metal-bearing DNA
oligomers for various applications.11,21
A strong hydrophobicity, or at least amphiphicity, is an inherent
property of most boron clusters used for the modification of
biologically important molecules. Surprisingly, in spite of
extensive biological and pharmaceutical research on boron-
cluster-containing conjugates, very little attention was paid to
the solution behavior of these molecules in aqueous media. It is
obvious that the aggregation process could reduce the concentra-
tion of active boron-containing molecules in the solution.
However, only very recent reports22-24 describe the strong
tendency of parental metallacarborane clusters to associate in
water. Furthermore, polymethylated dicarba-closo-dodecaborane
derivatives were designed as amphiphilic species that are able
to form supramolecular associates in water.25,26 The aggregation
of disaccharide conjugates with 1,2-dicarba-closo-dodecaborane
clusters was thoroughly studied in aqueous solutions.27-29
Because the incorporation of any drug into an aggregate
strongly affects its pharmacokinetics, interaction with biological
membranes,30 and biological activity in general,31,32 further studies
of the aggregation of boron-cluster-containing molecules are
needed. In this report, we demonstrate for the first time how the
size, charge, and exoskeletal substitution pattern of the boron-
containing cluster can influence the associative tendency of
modified nucleosides in aqueous solutions. We also suggest simple
rules for predicting the aggregation behavior of these compounds.
Experimental Section
Chemicals. 2′-Deoxyadenosine (dA) and thymidine (T) were
purchased from Pharma-Waldhof GmbH (Düsseldorf, Germany),
and ATP was purchased from Sigma-Aldrich (Sp. z o.o. Poznan,
Poland). 6-N-{5-[3-Cobalt(III) bis(1,2-dicarbollide)-8-yl]-3-oxa-
pentoxy}-2′-O-deoxyadenosine (1),33 and 5-(o-dicarba-closo-dode-
caborane-1-yl)-2′-deoxyuridine (CDU) (6)34 were obtained as
described previously. The syntheses of 8-[(p-dicarba-closo-dode-
caborane-2-yl)-ethynyl]-2′-deoxyadenosine (4), 2′-O-methyl-(1,2,3-
triazol-4-yl)-{4-N-{5-[3-cobalt(III) bis(1,2-dicarbollide)-8-yl]-3-oxa-
pentoxy}}-uridine (2), 3-N-propyl-(1,2,3-triazol-4-yl)-{4-N-{5-[3-
cobalt(III) bis(1,2-dicarbollide)-8-yl]-3-oxa-pentoxy}}-thymidine
(3), 8-{(1,2,3-triazol-4-yl)-4-N-{5-[(7,8-dicarba-nido-undecaborane)-
10-yl]-3-oxa-pentoxy}}-2′-O-deoxyadenosine (7), 2′-O-methyl-
(1,2,3-triazol-4-yl)-{4-N-{5-[(7,8-dicarba-nido-undecaborane)-10-
(20) Thirumamagal, B. T. S.; Zhao, X. B.; Bandyopadhyaya, A. K.;
Narayanasamy, S.; Johnsamuel, J.; Tiwari, R.; Golithly, D. W.; Patel, V.; Jehning,
B. T.; Backer, M. V.; Barth, R. F.; Lee, R. J.; Backer, J. M.; Tjarks, W. Bioconjugate
Chem. 2006, 17, 1141.
(21) Lesnikowski, Z. J. Curr. Org. Chem. 2007, 11, 355.
(22) Chevrot, G.; Schurhammer, R.; Wipff, G. J. Phys. Chem. B 2006, 110,
9488.
(23) Chevrot, G.; Schurhammer, R.; Wipff, G. Phys. Chem. Chem. Phys. 2007,
9, 1991.
(24) Matějı́ček, P.; Cı́gler, P.; Procházka, K.; Král, K. Langmuir 2006, 22,
575.
(25) Ma, L.; Hamdi, J.; Huang, J.; Hawthorne, M. F. Inorg. Chem. 2005, 44,
7249.
(26) Maderna, A.; Herzog, A.; Knobler, C. B.; Hawthorne, M. F. J. Am. Chem.
Soc. 2001, 123, 10423.
(27) Bonechi, C.; Ristori, S.; Martini, S.; Panza, L.; Martini, G.; Rossi, C.;
Donati, A. Biophys. Chem. 2007, 125, 320.
(28) Morandi, S.; Ristori, S.; Berti, D.; Panza, L.; Becciolini, A.; Martini, G.
Biochim. Biophys. Acta 2004, 1664, 53.
(29) Rossi, S.; Karlsson, G.; Martini, G.; Edwards, K. Langmuir 2003, 19,
5608.
(30) Schreier, S.; Malheiros, S. V. P.; de Paula, E. Biochim. Biophys. Acta
2000, 1508, 210.
(31) McGovern, S. L.; Helfand, B. T.; Feng, B.; Shoichet, B. K. J. Med. Chem.
2003, 46, 4265.
(32) McGovern, S. L.; Caselli, E.; Grigorieff, N.; Shoichet, B. K. J. Med.
Chem. 2002, 45, 1712.
(33) Olejniczak, A. B.; Plesek, J.; Lesnikowski, Z. J. Chem.sEur. J. 2007, 13,
311-318.
(34) Schinazi, R. F.; Goudgaon, N. M.; Fulcrand, G.; El Kattan, Y.; Lesnikowski,
Z. J.; Moravek, J.; Liotta, D. C. Int. J. Radiat. Oncol., Biol., Phys. 1994, 28, 1113.
Matějı́ček et al.
yl]-3-oxa-pentoxy}}-uridine (8), and 3-N-propyl-(1,2,3-triazol-4-
yl)-{4-N-{5-[(7,8-dicarba-nido-undecaborane)-10-yl]-3-oxa-pentoxy}}-
thymidine (9) were carried out using the “click chemistry” approach35
and will be described elsewhere. 2′-O-[(p-Dicarba-closo-dodecabo-
rane-1-yl)-propyleneoxymethyl]adenosine 5′-triphosphate (5) was
obtained from modified adenosine according to the general method
for nucleoside 5′-triphosphates synthesis.36 The countercation in
compounds 2, 3, 7, 8, and 9 was not unambiguously defined. Triton
X-100 was purchased from Merck KGaA (Darmstadt, Germany).
Methods. Preparation of Stock Solutions of ATP, T, dA, and
Compounds 1-9. All solvents and buffers were filtered using a 0.2
µm membrane filter before use. Stock solutions of 1-4 and 6-9
for DLS and SLS measurements were prepared in dimethylsulfoxide
(DMSO), diluted with water, and measured at a constant 0.5% (v/v)
DMSO concentration. Solutions of ATP, T, dA, and 5 were prepared
and measured in deionized water. In some cases specified below,
Triton X-100 was added to the solution so that the surfactant
concentration was either below the critical micellar concentration
cmc, (0.01%) or above it (0.25%).
Partition Coefficient (P) Measurement.37 Octanol (1 mL) was
added to a 100 µM solution of compounds 1-9 in deionized H2O
containing 5% CH3OH (1 mL) in a 2 mL Eppendorf tube. The
resulting mixture was shaken vigorously at room temperature (20-
22 °C) for 1 h to ensure that the compound’s transfer between the
two phases was at equilibrium; the mixture was then left standing
for 1 h in order to separate the phases. Each sample was subsequently
centrifuged for 5 min at 13 000 rpm, and then 0.9 mL of H2O or
organic solution was transferred to a 1 mL UV cell and the UV
absorption at λ ) 260 nm was measured. The partition coefficient
is defined as the ratio of the amount of compound present in the
organic phase to the amount present in the aqueous phase.
Dynamic Light Scattering (DLS), Static Light Scattering (SLS),
and Atomic Force Microscopy (AFM). The light scattering setup
(ALV, Langen, Germany) consisted of a 633 nm He-Ne laser, an
ALV CGS/8F goniometer, an ALV High QE APD detector, and an
ALV 5000/EPP multibit, multitau autocorrelator. DLS data analysis
was performed by fitting the measured normalized intensity
autocorrelation function g2(t) ) 1 + β|g1(t)|,2 where g1(t) is the
electric field correlation function, t is the lag time, and β is a factor
accounting for deviation from the ideal correlation. An inverse
Laplace transform of g1(t) with the aid of a constrained regularization
algorithm (CONTIN) provides the distribution of relaxation times,
τA(τ). Effective angle- and concentration-dependent hydrodynamic
radii, RH(q,c), were obtained from the mean values of relaxation
times, τm(q, c), of individual diffusive modes using the Stokes-
Einstein equation. To obtain true hydrodynamic radii, the data have
to be extrapolated to a zero scattering angle. To obtain information
on polydispersity, the analysis of the cumulant expansion of the
correlation function is performed by fitting a third-order polynomial
to the function ln(g2(t) - 1). The polynomial coefficients are converted
into the coefficients of the cumulant expansion of the field correlation
function g1(t). The polydispersity index (PDI) is calculated from the
second moment of third-order fitting. The PDI can be recalculated
in terms of polydispersity (PD) by the simple addition of unity. For
further information, see ref 38.
AFM measurements were performed in tapping mode under
ambient conditions using a Digital Instruments NanoScope Dimen-
sions 3 scanning probe microscope equipped with a Nanosensors
silicon cantilever. Details on the SLS, DLS, and AFM measurements
of the aqueous carborane solutions are given elsewhere.24
Results and Discussion
Light scattering measurements revealed important differences
in the association properties of investigated compounds in aqueous
(35) Kolb, H. C.; Finn, M. G.; Sharpless, K. B. Angew. Chem., Int. Ed. 2001,
40, 2004.
(36) Mishra, N. C.; Broom, A. D. J. Chem. Soc., Chem. Commun.1991, 1276.
(37) Dagle, J. M.; Andracki, M. E.; DeVine, R. J.; Walder, J. A. Nucleic Acids
Res. 1991, 19, 1805.
(38) Forster, S.; Zisenis, M.; Wenz, E.; Antonietti, M. J. Chem. Phys. 1996,
104, 9956.
Aggregation of Boronated Nucleosides
solutions in the concentration range of 5.0 × 10-7 to 1.0 × 10-4
mol/L. It is worth mentioning that 1-6 form relatively large
aggregates, whereas the other compounds (7-9, unmodified
nucleosides T and dA, and nucleotide ATP) did not form any
aggregates large enough to scatter visible light significantly. The
mean hydrodynamic radii RH of particles formed by the individual
compounds are listed in Table 1, together with relative light
scattering intensities and polydispersity indexes of RH distribu-
tions, both measured at a 90° scattering angle (where the scattering
intensity of the solvent is set equal to unity). It is obvious that
the species capable of forming the aggregates contain either the
bulky [3-cobalt(III) bis(1,2-dicarbollide)]- ion (compounds 1-3)
or the electroneutral dicarba-closo-dodecaboranyl moiety (com-
pounds 4-6).
Typical distributions of RH for aggregates obtained by DLS
are shown in Figure 1a,b for the both [3-cobalt(III) bis(1,2-
dicarbollide)]- ion-based and dicarba-closo-dodecaborane-based
compounds, respectively. The aggregates of 1 and 4 are fairly
monodisperse in dilute solutions. Their weight fractions are quite
high (on the basis of high intensities of scattered light), whereas
the weight fractions of 2, 3, 5, and 6 are on the edge of LS
detection. (See the LS intensities in Table 1.) The size of the
aggregates corresponds reasonably with the radii of 1,2-dicarba-
closo-dodecaborane cluster-disaccharide conjugates in dilute
solutions obtained by DLS as published by Morandi et al.28 Similar
values were also found for the parental metallacarborane cage
anions.24
As published earlier,24,28,29 the behavior of boron cage
aggregates can change with concentration. In accordance with
this, the experimental hydrodynamic radii and polydispersity of
compounds 1 and 4 depend strongly on their concentration. 4
forms relatively small, monodisperse aggregates in dilute solution
(RH ca. 140 nm at 10-6 mol/L). However, at concentrations larger
than 10-5 mol/L, secondary aggregation accompanied by partial
precipitation occurs. Dilute solutions of 1 (c < ca. 5 × 10-6 M)
contain an insignificant weight fraction of relatively small
particles, but above this limit, fairly monodisperse and quite
stable particles form. The size distribution of aggregates formed
by boron-modified ATP (5) is of interest. The experimental
distribution is distinctly bimodal (Figure 1b), which is an
interesting observation requiring additional study.
We point out that it is difficult to obtain reliable aggregation
numbers in dilute solutions of only partially aggregating low-
molar-mass compounds by LS measurements without information
provided by other experimental methods. However, a rough guess
concerning the apparent weight-average molar mass can be based
on a combination of RH and I rel/c values (Figure 2), where I rel
is the relative light scattering intensity extrapolated to zero
scattering angle and c is the analytical concentration.
In spite of the fact that the experimental average RH values
of compounds 2, 3, 5, and 6 are not negligible, the I rel values
are very low, indicating that the concentration of aggregated
molecules is low. However, Figure 2 shows that I rel/c values do
not change significantly with concentration. It suggests that the
aggregation number does not change with c and the fraction of
aggregates is proportional to the concentration within the range
of the experimental errors. Assuming that the aggregates appear
at a certain concentration (ca. 1 × 10-5 M) and the diameters
of the aggregates as well as the aggregation numbers change
very little with the concentration, the behavior of the compounds
seems to obey the mechanism of the closed association.
Nevertheless, a fraction of the aggregated molecules above the
apparent critical aggregation concentration (cac) is very low.
Langmuir C
In the case of solutions 1 and 4, the situation is different. The
dicarba-closo-dodecaborane-modified 2-deoxyadenosine (4) tends
to aggregate significantly even in very dilute solutions and
eventually starts to precipitate at concentrations of ca. 1 × 10-5
M. Because the values of I rel/c and RH are nearly unchanging
with c up to precipitation, it seems that the concentration of the
nonaggregated form remains roughly the same. In other words,
the apparent cac is below our limit of detection, and the size of
the aggregates is thermodynamically controlled below the
precipitation point.
The system containing 1 forms only an insignificant number
of nanoparticles, indicating that preaggregation takes place below
a concentration of 5 × 10-6 M. Above this limit, the fraction of
aggregates increases considerably. This concentration can be
considered to be the apparent cac, although the aggregation
number and the concentration of nonaggregated molecules seem
to change above the cac (curve 4 in Figure 2). Unfortunately we
cannot obtain the ratio of the aggregated and nonaggregated
molecules (1-6) by the LS measurements.24 Thus, these
conclusions are only on the qualitative level.
From the LS measurements, it is evident that solutions of
compounds listed in Table 1 contain aggregates; however, a
detailed study of their structure requires more direct experimental
techniques (e.g., atomic force microscopy, AFM). Although the
shape of colloidal particles is often deformed after their deposition
on the substrate, we have shown that the original size and shape
of stable nanoparticles in solution correlates with those on the
surface.24 A typical scan of particles formed by 1 deposited on
flat mica surfaces is shown in Figure 3a. We can assume from
the roughly round stains, which correspond to the aggregates,
that the particles could be spherical in solution. The AFM
technique was also used to image deposited 4 and 5. The scans
for 4 are very similar to those for 1 (not shown). The AFM
results verify the LS observations that particles formed by 5 are
very polydisperse (Figure 3b).
The aggregation behavior of the studied compounds can be
explained on the basis of boron-cluster properties and the overall
polarity of individual molecules. The dicarba-closo-dodecabo-
ranyl moieties are electroneutral and significantly more hydro-
phobic than the negatively charged dicarba-nido-undecaboranyl
ones. The [3-cobalt(III) bis(1,2-dicarbollide)]- cluster bears a
negative charge, but it retains strong hydrophobic character
because of its bulkiness. In light of the above consideration, one
can classify the individual compounds and rationalize the observed
behavior.
However, the hydrophobic character of all of the mentioned
clusters is not strong enough to cause the quantitative aggregation
of polar nucleosides, and the solutions of 2, 3, 5, and 6 contain
only a small fraction of the aggregates. Concerning 5, the tendency
to form a large aggregate is counterbalanced by the presence of
the hydrophilic triphosphate unit. This compound is strongly
amphiphilic and its behavior is peculiar, as evidenced by both
LS and AFM measurements. The distinctive propensity of 1 to
aggregate is probably due to the well-known interaction of the
[3-cobalt(III) bis(1,2-dicarbollide)]- cluster with amino groups,
resulting in sparingly soluble species.39,40 The amino group of
adenosine is a typical candidate for the formation of zwitterionic
species of this type. Moreover, we observed that all [3-cobalt-
(III) bis(1,2-dicarbollide)]--containing compounds are almost
insoluble in buffers containing molecules bearing amino groups
that are currently used in biology-oriented studies (such as tris-
(hydroxymethyl)aminomethane, TRIS).
(39) Plesek, J.; Base, K.; Mares, F.; Hanousek, F.; Stibr, B.; Hermanek, S.
Collect. Czech. Chem. Commun. 1984, 49, 2776.
(40) Plesek, J. Chem. ReV. 1992, 92, 269.
D Langmuir Matějı́ček et al.

Table 1. Characterization of the Aggregates in Solution by Light Scattering (Mean Hydrodynamic Radius RH, Relative Scattering
Intensity and Polydispersity Index (PDI) for a 90° Scattering Angle) and the Partition Coefficient (P) for Nucleoside-Boron Cluster
Conjugates 1-9 and Unmodified Nucleoside Counterparts

*For details, see Experimental Section (Chemicals). a The light scattering intensity is too low to obtain reliable data. b The solution does not contain
particles capable of scattering the light within the studied concentration range (5.0 × 10-7 to 1.0 × 10-4 mol/L). c The particles are very polydisperse
or contain a small fraction of very large aggregates. d The solution slowly precipitates. e Reference 17. f Drug Bank data: log P ) -1.08. g Drug
Bank data: log P ) -0.72. h Drug Bank data: log P ) -5.5. i Calculated44 (VCCLAB, http://www.vcclab.org): log P ) -1.23. j Calculated44
(VCCLAB, http://www.vcclab.org): log P ) -1.35.
Aggregation of Boronated Nucleosides Langmuir E

Figure 1. Typical distributions of hydrodynamic radii, RH, of (a)

1 (-), 2 (---), and 3 (‚‚‚) and (b) 4 (-), 5 (---), and 6 (‚‚‚) at a 90°
scattering angle. The concentrations of the individual species were
as follows: 3.16 × 10-6 M for 4, 1.78 × 10-5 M for 1, and 1.00
× 10-4 M for 2, 3, 5, and 6.

Figure 3. AFM scan on a mica surface for 1 deposited from the

solution with a concentration of 5.62 × 10-6 M (a) and 5 deposited
Figure 2. Relative light scattering intensity extrapolated to a zero from the solution with a concentration of 1.00 × 10-5 M (b).
angle divided by concentration, I rel/c, as a function of concentration
for compounds 1-6. applications. Therefore, we carried out preliminary light scattering
experiments with nonionic surfactant Triton X-100, which forms
micelles with a radius of ca. 3 nm. It is well known that
These results of LS and AFM studies can be compared with
hydrophobic compounds can disperse in the surfactant micelles.
a simple and (in biomedical sciences) commonly used charac-
Stable solutions of 1-4 and 6 were studied in the presence of
terization of the compound’s hydrophobicity based on the partition
Triton X-100 at a concentration of 0.25% by LS. This is well
coefficient P between water and octanol. Furthermore, P is useful
above the cmc (ca. 0.013%). 1 was dispersed in the surfactant
for potential biological and pharmaceutical applications and
micelles quantitatively as indicated by a significant decrease in
application-oriented studies. The results are summarized in Table
the LS intensity and the diminishing of the 1 peak in the RH
1 for all studied species together with their structural formulas.
distribution. Within the uncertainty due to low LS intensities,
The P values correlate fairly well with the aggregation tendency
the same is also true for 2, 3, and 6. The only peculiar exception
observed by LS. 1 and 4, which exhibit pronounced association
is 4, where the peak corresponding to the aggregate did not
in aqueous solutions, have quite high P values. The values for
disappear in the DLS experiment. However, other LS charac-
2 and 3 are moderate but still higher than unity. 6 seems to be
teristics such as the peak position and the overall LS intensity
a peculiar exception. It aggregates only weakly in spite of the
changed in the presence of surfactant micelles, indicating mutual
high value of P. However, the assumption that compounds with
interaction. Interactions of the studied compounds with the Triton
P > 1 form aggregates in the aqueous solutions is still fulfilled.
X-100 molecules were also observed in solutions below the cmc
5 and the rest of the studied compounds exhibit P < 1. This is
of Triton (0.01%). The radii of large nanoparticles of 1 and 4
due to the presence of a highly hydrophilic triphosphate moiety
decreased to ca. 105 and 75 nm, respectively, though the fractions
in 5, which makes the molecule more hydrophilic and lowers its
of the aggregated molecules remain quite high. None of these
solubility in octanol. It is a very useful finding that, at least for
observations are surprising when we assume that the considered
the series of studied compounds, partition coefficients (which
boron cluster compounds exhibit amphiphilic and surface-active
can be routinely measured) predict aggregation in aqueous
behavior22-24 and they are also predisposed to form double
solutions.
hydrogen bonds.43 The obtained results are so complex and
However, it should be stressed that although aggregators, in
interesting that the interaction of boron-containing compounds
general, often have higher P values than molecules that do not
aggregate, this criterion by itself must be used cautiously because (41) Seidler, J.; McGovern, S. L.; Doman, T. N.; Shoichet, B. K. J. Med.
in many cases it may not be sufficient to robustly distinguish Chem. 2003, 46, 4477.
molecules with different tendencies to aggregate.41,42 (42) Feng, B. Y.; Simeonov, A.; Jadhav, A.; Babaoglu, K.; Inglese, J.; Shoichet,
B. K.; Austin, C. P. J. Med. Chem. 2007, 50, 2385.
The fact that some boron-containing conjugates form ag- (43) Fanfrlik, J.; Lepsik, M.; Horinek, D.; Havlas, Z.; Hobza, P. ChemPhysChem
gregates in solution can deteriorate their properties in potential 2006, 7, 1100.
F Langmuir PAGE EST: 5.3
with the amphiphilic systems requires a separate extensive study,
which will be published soon.
In summary, we report the first observations concerning the
behavior of nucleosides substituted by boron clusters in aqueous
media. The combined LS and AFM study revealed an intricate
balance between hydrophobic and hydrophilic interactions and
the sensitivity of the behavior to the nature of boron clusters:
nucleosides with attached electroneutral or bulky charged clusters
aggregate spontaneously, whereas nucleosides containing smaller
and charged nido clusters as well as the unmodified nucleosides
remain in the form of true solutions or oligomolecular associates
that are not observable by LS. The aggregation is strongly
enhanced by the presence of amino groups because of the
formation of zwitterions, which are sparingly soluble in the
aqueous solution. We demonstrate that the partition coefficient
P enables the prediction of the association of boron-cluster-
containing nucleobases on a qualitative level.
(44) Tetko, I. V.; Gasteiger, J.; Todeschini, R.; Mauri, A.; Livingstone, D.;
Ertl, P.; Palyulin, V. A.; Radchenko, E. V.; Zefirov, N. S.; Makarenko, A. S.;
Tanchuk, V. Y.; Prokopenko, V. V. J. Comput.-Aided Mol. Des. 2005, 19, 453.
Matějı́ček et al.
Acknowledgment. This article is dedicated to Professor
Jaromı́r Plešek on the occasion of his 80th birthday in recognition
of his outstanding contributions to the areas of borane and
carborane chemistry. We express our gratitude to Dr. Milena
Špı́rková (Institute of Macromolecular Chemistry, Academy of
Science of the Czech Republic in Prague) for AFM measurements.
Software provided by www.nanotec.es was used for the imaging
of AFM scans. This work was supported in part by the Polish
Ministry of Science and Higher Education (grant no. N405 051
32/3592 to A.B.O., A.A., B.W., and Z.J.L.), the Grant Agency
of the Czech Republic (grant no. 203/06/P138 to P.M.), the Marie
Curie Research and Training Network (grant no. 505 027,
POLYAMPHI to P.M. and K.P.), and a grant from the Ministry
of Education (MSMT) of the Czech Republic within program
1M6138896301 (“Research Centre for New Antivirals and
Antineoplastics”) to P.C. P.M. and K.P. were supported by a
long-term research plan of the Ministry of Education of the Czech
Republic (grant no. 0021620857).
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