Medicinal applications of fullerenes : A literature review

Patrycjusz Cieślak

Introduction

Fullerenes (family of carbon allotropes) discovered in 1985 have many interesting

properties. Synthesized hydrophilic fullerene derivatives enable research in medical
applications of those molecules. C60 molecules can be used as inhibitors of HIV protease1.
Fullerene derivatives applied into DNA or lipid membranes exhibit interesting photochemical
properties. Exposed to light, fullerenes can produce singlet oxygen and can be used in cancer
therapy. Some fullerene derivatives are able to neutralize free radicals enabling usage of
fullerenes as antioxidants. In this review, I want to delve into the usage of fullerenes in
medicine, their biological and biochemical aspects, that have already been introduced.
Fullerenes are - besides graphite and diamond - the third allotropic variant of carbon.
This name covers the whole family of particles with the general formula C 2n (n > 16) in which
the carbon atoms are located only on the surface of the body. Among the large family of
fullerenes, the most common, and at the same time the best known, are the fullerenes
containing 60 or 70 carbon atoms. These particles show many interesting properties and their
electrochemical properties are particularly interesting and intensively studied. Multiatomic
carbon fullerene solids are completely empty inside and have an internal space large enough
to fill even the largest chemical element atoms. To date, researchers have been able to place
multidimensional metal atoms, including radioactive elements and noble gas atoms, inside the
carbon sphere. The most popular among scientists is the Fullerene C60 particle, because due
to its spatial structure it is considered to be "ideal". Fullerene particle C60 has the shape of a
spheroid, or more precisely, a truncated molded icosahedron, which has 60 vertices, each
being one carbon atom.
The main problems concerning the use of fullerenes in medical chemistry were
insolubility in polar solvents and forming aggregates in aqueous solvents. However, these
problems were solved by a number of chemical modifications of fullerene molecules.

History of fullerenes

Harold Kroto from the University of Sussex is considered to be the pioneer of the
discovery of fullerenes. In his PhD thesis he investigated the transformations of carbon
compounds taking place around extinct stars using spectroscopic methods and discovered
characteristic narrow spectral lines that corresponded to aromatic carbon compounds.
For the discovery of the Harold Kroto fullerenes from the University of Sussex in
Brighton (UK) and the team of R.E. Smalley and R.F. Curl jr. from the University of Rice in
Huston (Texas, USA), they received the 1996 Nobel Prize in chemistry. Harold Kroto
continued his research on fullerenes at the University of Sussex, including extracting them
from soot in a very time-consuming way and began to study their chemical properties. In
1990, German researchers W. Kratschmar and D. Huffman published for the first time a
relatively cheap and efficient method of fulerene synthesis through controlled burning of
carbon in an electric arc in a helium atmosphere, which opened the way for the practical
application of these compounds, but was not included in the Nobel Prize.
 In memory of the Dallas dome designer, they started to call each other "Bucky balls"
in jokes of this type of relationship (that is, in free translation "Bucky's testicles"), which in
the first publication was transformed into the more serious-sounding name "Buckminster
fulleren", from which the name for the whole group of compounds was invented.

Construction of fullerenes

C60 is made of 20 hexagonal walls and 12 pentagonal walls. Each of the 20 hexagons
borders on three pentagons and three hexagons. Each of the carbon atoms of Fullerene C60
has the same environment, so all the atoms are equal and the molecule does not contain weak
points of chemical interactions. Although the folding of the graphite layer into a carbon cage
is accompanied by stresses, due to symmetry they are evenly distributed throughout the
particle.
The special structural isomers of fullerenes are nanotubes, which are long cylinders
obtained from coiling a single graphite plane, closed on both sides with halves of fullerenes of
appropriate size. The shortest nanotube, from a formal point of view, is C70, while the longest
(state of the art in 2008) is over two centimetres long.
The IUPAC method of nameing Buckminsterfullerene, before it was finally
announced as C60, was too lenghty and complicated for general use:
Hentriacontacyclo[[Link].[Link].04.59.05,10.06,58.07,55.08,53.09,21.011,20.011,20.013,18.015,30.016,28
.017,25.019,24.022,52.023,50.026,49.027,47.029,45.032,44.033,60.034,57.035,43.036,56.037,41.038,54.039,51.040,48
.042,46]hexaconta-1,3,4(10),6,8,11,16(18,14,16,19,21,23,25,27,29(45),30,32,(44),33,35(43),36,
38(54),39(51),40(48),41,46,49,52,55,57,59-triacontaene2.

Application of fullerenes

The use of fullerenes and their derivatives Fullerenes are increasingly used primarily
in diagnostics and medicine, as well as in the electronics and energy industries. They show
potential antiviral effects, e.g. as inhibitors of aspartic proteases of the human
immunodeficiency virus (HIV), fitting into the centre of the active enzyme and interacting
with the van der Waals bonds. Moreover, fullerenes have a high antioxidant potential.
Already 1 molecule can neutralize more than 20 free radicals. The antioxidant properties are
caused by a large number of conjugated double bonds and a low lying unfilled molecular orbit
that can absorb the electron. This gives a chance to counteract oxidative stress and oxidative
damage to cells3,4. Fullerenes and their water-soluble derivatives can be used as radiation
protection factors in radiotherapy. They prevent DNA damage caused by ionizing radiation 5.
 Researchers are working on the possibility of using fullerenes as factors protecting
nerve cells from excess free radicals in neurodegenerative diseases such as Alzheimer's,
Parkinson's or ALS (atrophic lateral sclerosis). Fullerenes can also prevent cell apoptosis.
Research is ongoing on their potential use in other diseases such as atherosclerosis and
osteoporosis. Fullerenes can also protect cells from the harmful effects of UVA radiation.
They are also the subject of research in photochemical therapy, especially in the treatment of
skin cancer, thanks to the possibility of cutting the oligonucleotide chain after photon
excitation, which leads to the transformation of molecular oxygen into the 1O2 singlet excited
state. According to some researchers, fullerenes can be used as contrast agents in diagnostics.
It is also possible to use these particles, interesting in terms of structure and function, as
potential drug transmitters or gene carriers3, 4.

Toxicity of fullerenes and its derivatives

The absorption and distribution of fullerene is highly dependent on the route of
exposure. In inhaled animals, fullerene was mainly detected in different sections of the
respiratory system. From the tracheobronchial and bronchial sections, the nanoparticles were
removed together with mucus through the movements of ciliae of the epithelium. The
particles, which reached the lungs, were phagocytised by alveolar macrophages. Fuleren was
slightly absorbed from the gastrointestinal tract after intragastric administration, most of the
administered dose was excreted with faeces. No skin absorption was observed6.
In inhalation studies on rodents (mainly rats), fullerene caused temporary
inflammatory changes in the lungs. The inflammation was manifested by an increase in the
concentration of proinflammatory proteins in broncho-alveolar lavage fluid (BALF).
Histopathological studies also revealed the formation of inflammatory cell infiltration in the
lungs, cell proliferation or collagen fibrosis of the bubble walls; however, these changes
subsided one month after the exposure stopped. The oral exposure did not cause major
adverse effects7.
Fluorescence of Fullerene
Fluorescence properties of fullerenes and its derivatives are of considerable interest to
chemists and spectroscopists. Because C60 contains thirty conjugated double bonds, in the
UV-VIS [60]fullerene spectrum many absorption bands corresponding to electron transitions
S0→ Sn are observed. Fluorescence emission from fullerene is also expected, because the
structural stiffness maintained by double interlocking bonds usually favours deactivation of
the excited state by radiation emission rather than by internal conversion.
Despite extensive research, previous studies have not allowed the detection of
fullerene fluorescence at room temperature, probably due to the very low quantum
fluorescence yield. Fluorescence at room temperature was first detected for fullerene C 70. In
addition, recent studies have observed that the asymmetry of fullerene structure increases with
the increase of fluorescence in C60 derivatives at room temperature8.
The use of water-soluble, non-toxic fullerene derivatives arouses considerable interest
in relation to current research on the creation of nanomedical drug carriers in modern
anticancer therapies. The chemical modifications of [60]fullerene described earlier in the
scientific literature, including Bingel-Hirsch reactions, Prato reaction and fullerene
cycloaddition reactions, allow to conveniently create covalent connections with many drugs in
order to better deliver the drug to the molecular target9.
Carbon nanomaterials can now be used to determine biomolecules in cells to observe
the progression of disease and can also be introduced into living cells as contraindications10.

There are two distinct unimolecular mechanism for molecular fluorescence: prompt
fluorescence (PF) and thermally activated delayed fluorescence (TADF). Everything is based
on their mechanisms. In the PF – the emission occurs after Sn  S0 absorption and excited
state relaxation to S1. In contrast, TADF mechanism takes place by way of triplet manifold:
after excitation and once S1 is fulfilled, there is an intersystem move to the triplet manifold.
There is a possibility, that there is a move back to S1, before the fluorescence occurs. The
sequence S1T1S1 may be repeated several times before fluorescence finally takes place.
TADF is especially important only when the quantum yield of triplet formation and the
quantum yield of singlet formation are both high. This outcome in turn implies a small energy
gap between S1 and T1, a long T1 lifetime, and not too low a temperature. For a given
fluorophore, TADF is usually much weaker than its PF. Although known for many years,
TADF continues to be a rare phenomenon, with a few observations in some xanthene dyes,
aromatic ketones and thiones, metal porphyrins, and aromatic hydrocarbons11.

Water – soluble C60 fullerenes in anti-cancer therapies

Medical chemistry research on carbon nanomaterials carried out in the last 20 years
confirms that fullerene derivatives of C60 and C82 can inhibit the growth of malignant tumours.
Such behavior can be explained by high antioxidant activity and by blocking special receptors
such as epithelial growth factor12.
Thanks to a large number of synthetic methods allowing for strictly defined
functioning of fullerene C60, they can be used as convenient drug delivery systems, especially
for modern anticancer therapies. Thus obtained fullerene nanomaterials are located both in the
intracellular complex of nuclear pores and blood vessels of cancer cells13.
Moreover, recent studies on sugar derivatives of fullerene (glycofullerenes) describe
their phototoxic effects in pancreatic cancer cells. This is because cancer cells have an
increased need for glucose, which is metabolized to a much greater extent in order to obtain
energy, which is needed for the reproduction and increased activity of cancer cells. In
addition, the family of transporting proteins is largely prevalent in many types of cancers 13.

Antioxidant properties of fullerenes

The antioxidant properties of water-soluble [60]fullerenes have been described earlier
in numerous reports and are based on the fact that fullerenes have in their structure a large
number of double bonds with very low level of the lowest unfilled molecular orbits (LUMO),
which can easily absorb the electron. Due to the ability to locate in cellular mitochondria, as
well as in other parts of cells where the disease spreads, fullerenes have gained considerable
popularity in use as medical antioxidants14.
When fullerenes have been functionalized by adding a strong polar group, as with
polyhydroxyl fullerenes, they can penetrate human cells. In addition, due to their high water
solubility, they can pass through cell membranes and locate in the mitochondria, which can
generate large amounts of free radicals. This has made fullerenes useful in many branches of
medicine. These molecules have shown that they are excellent at counteracting many toxins,
which can cause a lot of damage to many types of cells, such as nerve cells, liver cells and
epithelial cells4.
Fullerenes are also used as cellular protection against UVA radiation. Ultraviolet radiation
generates oxidative units that have a clear effect on human skin cells, which can lead to
damage or even death. Once again, water-soluble fullerenes were used to protect human cells
or other mammals from oxidative stress15.

Conclusion

Fulerenes have many interesting properties. Due to the presence of delocalized

electrons they can take and give off electrons and exhibit both prooxidative and antioxidative
properties. Fullerenes can induce the conversion of molecular oxygen into a peroxide anino
radical. Under the influence of visible light, hydroxyl radical ( -OH) and superoxide anino
radical (O2-) may be formed in polar solvents and in non-polar solvents singlet oxygen (O2).
The controlled release of these reactive oxygen species gives a wide range of applications for
fullerenes, including photodynamic cancer therapy. On the other hand, water-soluble fullerene
derivatives, including hydroxyl substituted fullerols, show antioxidant properties and can act
as free radical scavengers (they are even called "radical sponges"). The activity and properties
of these molecules largely depend on substituents and even on the solvent itself . The
toxicokinetics of fullerenes, i.e. their absorption, distribution and excretion, also depends on
the properties of particles, substituents and even the solvents used. Fulerene C60 is
characterized by poor absorption and low toxicity. It does not pose a hazard in the working
environment.

The exponential increase in patent filing and publications indicate growing industrial interest
that parallels academic interest. The discovery of fullerenes has been compared to the
discovery of benzene by many researches. The hydrophobic spheroid structure and radical
sponge character of the fullerene molecule is responsible for activities in many fields. In
addition to some of the important medical applications as mentioned above, fullerenes are
being intensively investigated for practical utilization in a number of other fields, e.g., as
room temperature superconductors, as component of solar batteries, for production of
synthetic diamonds and as lubricants.

However, there are two important facts that are considered to be barriers to fullerene
applications. The first one is relative insolubility and instability of fullerenes to water. This
however is being successfully overcome with the development of numerous water-soluble
fullerene derivatives. The second and more important limitation is the high cost of fullerenes.
Compared to gold that is available at the cost of $10/g, the cost of fullerenes, as listed on web,
varies from $45 to $1000/g16, depending on degree of purification. This high cost, which is
mainly attributed to very high temperature involved in the production process, is restricting
the numerous potential applications of fullerenes. One possible development that may take
place in near future is creation of modest demand of fullerenes in medicine, as medicine is
considered to be the cost-insensitive application. This in turn will encourage larger production
leading to lower prices and still more demand. The preliminary biological investigations of
fullerene derivatives as discussed above do promise encouraging results. Hence research
efforts in both chemistry and biology should be to further investigate the potential
applications of these interesting spheroidal molecules.
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Lang=pl&region=PL.