See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/281935258

Comments to the Mechanism of Protective and Pharmacological Action of

Radioprotectors from the Family of Aminothiols

Article · March 2014

DOI: 10.12966/jrr.06.03.2014

CITATIONS READS

3 326

1 author:

Mikhail Vitalyevich Vasin

Russian Medical Academy of Continuous Professional Education, Moscow
142 PUBLICATIONS   336 CITATIONS   

SEE PROFILE

All content following this page was uploaded by Mikhail Vitalyevich Vasin on 20 September 2015.

The user has requested enhancement of the downloaded file.

Sciknow Publications Ltd. JRR 2014, 2(2):15-36
Journal of Radioprotection Research DOI: 10.12966/jrr.06.03.2014
©Attribution 3.0 Unported (CC BY 3.0)

Comments to the Mechanism of Protective and Pharmacological

Action of Radioprotectors from the Family of Aminothiols
Mikhail V. Vasin*
Department of M edicine of catastrophe, Russian M edical Academy of Post -graduated Education, M oscow, Russian Federation

*Corresponding author (Email: mikhail-v-vasin@yandex.ru)

Abstract - This review evaluates a characteristic features of the mechanism of radiation protective action of aminothiols hence
their pharmacological properties and their difference fro m the rest of antio xidants. Over the years, extensive experimental stud-
ies of radiation protective agents from aminothiols series were completed by discovery of radioprotectors cystamine and
amifostine used in medical p ractice. Characteristic features of the mechanism o f radiation protective act ion of aminothiols un-
like other antio xidants are potential chance of the development of comp lete neutralizat ion of o xygen effect as radiological
phenomenon, first of all, in DNA radio lysis. This peculiar action of aminothiols is connected with their chemical structure be-
ing able to squeeze DNA helix v ia diamine bond of aminothiol disulfide thereby decreasing availabilility of crit ical points o f
DNA mo lecules to radicals of oxygen species. Positive charge of aminothiol molecules has an advantage for the development
of high their concentration in negative DNA mo lecules. In addit ion, pharmacological properties of aminothiols can deierminate
complex polyfunctional action that is significant for the radiation protection, including the in fluence to mitochondria respira-
tion follo wing acute hypoxia, gene Mn-SOD expression, activity of the Tip60 acetylransferase, etc. However, the decrease of
DNA damage by aminothiols is basic factor in their p rotective action. The properties of aminothiols t o block transaminase fol-
lowing a develop ment of hypotension expand their application in clinical practice and the same thing is a cause for severe side
effect. Character of aminothiol pharmad inamics is necessary to take into consideration in its repeated a pplication. The scheme
of prophylactic application of aminothiols in radiotherapy and for potential victims of radiation accidents is analy zed.
Keywords - Aminothiols, Radioprotectors, Aminofostine, Cystamine, Radiation Protection, Pharmacodinamics

1951), ethanol (Paterson et al., 1951), sodium nitrite (Co le et

1. Introduction. History of Discovery of
the Mechanism of the Action of Ra-
dioprotectors
The possibility of reducing the radiation damage in
mammals by using sulfur containing compounds was first
demonstrated in the studies of the effects of the cysteine
administration by Patt et al.in 1949, on the examp le of
glutathione by Chapman et aland thiourea by Mole et al. and
Limperose et al. in 1950. By that time, the “o xygen” effect,
the universal radiobio logical phenomenon manifested in the
increased radiation damage to organic substances and the
liv ing organism as a whole in the presence of oxygen in the
environment had been discovered (Giles & Riley , 1950;
Limperose, 1950; Do wdy, 1950; Gray et al., 1953; Alper &
Howard-Flanders, 1956). It is noteworthy that the radiation
protective effect was at the same time found in the
pharmacological co mpounds which at certain doses caused a
hypoxic state of the organis m: cyanide (Bacq et al., 1950), n -
amino-propiophenone (Storer et al., 1950), mo rphine (Kahn,
al., 1952), serotonin, adrenaline, and hystamine (Gray et al.,
1952a,b; Bacq et al., 1952; van den Brenk & Elliot, 1958).
Studies by van den Brenk et al. (1959, 1962) first
demonstrated the possibility of elimination or a substantial
reduction of the radiation protective effect of adrenaline,
histamine, serotonine and aminothiols under the influence of
the high oxygen pressure.
In 1951, Z. Bacq discovered radioprotector cysteamine
and its disulfide - cystamine, which attracted the widespread
attention of radiobiologists because of their high act ivity that
could potentially find some practical applicat ions and be
used to protect humans against the effects of ionizing
radiation. Systematic long-term studies using cysteamine,
which were aimed at the discovery of new effective
radioprotectors from the group of aminothiols, were
launched by a number of large scientific centers and
laboratories around the world. In the subsequent years, the
search for new radioprotectors from d ifferent classes of
chemical co mpounds was intensively conducted. 2-A mino-
ethyl isothiuronium (A ET) (Doherty & Burnett, 1955;
Langendorff & Koch, 1956; Shapira et al., 1957), 2-
16 Journal of Radioprotection Research(2014) 15-36
aminoethylthiazoline (Shapira et al., 1957), some
dithiocarbamates (Van Bekku m et al., 1956), cysteamine S-
substituted compounds - 2-amino-ethyl thiosulfuric acid
(Holmberg et al., 1959, Kaluszyner et al., 1961) and the most
interesting of them - 2-amino-ethyl thiophosphate (WR-638)
(Akerfeldt et al., 1959, 1963, 1967; Hanson & Sorbo, 1961)
were equal in the effectiveness with cysteamine.
Cystamine, radioprotector fro m the group of
aminoalkylthiols represents itself a d isulfide of -
mercaptoethylamine (M EA ) or cysteamine. When entering
the blood, cystamine is rapid ly non-enzy matically reduced to
MEA and imp lements its radiation protective effect with the
participation of a free thiol group in the compound:
H2 N–(CH2 )2 –S–S–(CH2 )2 –NH2  2 H2 N–(CH2 )2 –SH
Radiat ion protective properties of sulfur containing
compounds are sufficiently increased in the p resence of the
mo lecule of the amino group in the structure. The amine
group can be replaced by the amid ine group. In this group,
the electron density on the nitrogen atoms increases, which
contributes to the effectiveness of radioprotectors.
2-A minoetilizotiuroniu m (A ET), an active radioprotector
which has a protective effect at super lethal doses of
radiation exposure, undergoes intramo lecular rearrange ments
(“transguanilation”) to mercapto-ethylguanidine in aqueous
milieu at pH = 7, i.e., under physiological conditions.
Transformat ion into the co mpounds with a free thiol group is
important for the manifestation of radiation protective
properties of sulfur containing hereto cyclic co mpounds:
thiazolines and thiazolid ines (Shapira et al., 1957). The
intermediate product of the rearrangement of isothiuronium
is 2-aminotiazo lin. Th iazo lid ines can be considered a
deposited form of M EA for their ability to split in the body
with the release of the latter substance. Apparently, this is
the reason for why they have a protective action when
administered orally.
Reducing properties of sulfur in the mo lecules of
dithiocarbamates also predetermine their radiation protective
activity (van Bekku m, 1956). They are closely related to
isothiuronium co mpounds in their structure. The ability to
bind and inactivate the hydroxyl radical, the most active
product of the radiolysis, predetermines the radiation
protective properties of dimethyl sulfo xide (СH3 )2 S=O
(Ashwood-Smith, 1961). The necessary conditions required
for the imp lementation of radiation protective properties of
sulfur containing radioprotectors include their mandatory
delivery into the cell and accumu lation in the particular sub-
cellu lar structures. The pharmacological drugs which fail to
penetrate into cells or those with a lo w rate of transition
through cellu lar membranes do not have the radiation
protective effect.
First tests of the most active rad ioprotectors (cystamine,
cysteamine, A ET, serotonin and mexamine) had been carried
out by this time on large animals (dogs and monkeys)
(Crough et al., 1957; Jacobus et al., 1959; Razorenova et
al.,1961; Benson et al., 1961; Semenov et al., 1963). They
showed that radiation protective agents had radiation
protective effects on large animals; however, the observed
activity of the pharmaco logical preparations was more
moderate as co mpared with the results obtained on small
laboratory animals and, as a rule, d id not exceed 50% for
sulfur-containing compounds and 20 - 30% in the testing of
serotonin and mexamine.
The history of the discovery of rad iation protective
compounds is closely related to the ach ievements in the field
of radiation chemistry of that time. The direct evidence for
the formation of the hydro xyl radical as a result of the effects
of ion izing radiation on water was obtained by Weiss in
1944. In 1947, studies of Lea provided the first exp lanation
of the mechanism for the formation of radicals fro m ions as a
result of their reco mb ination in the course of radiolysis.
Barron (1949) was the first who observed reparation of
irradiated macromolecules on the example of sulfhydryl
enzymes in the experiments in vitro using low molecular
weight SH - compounds. This research into the indirect
effects of radiation in biological systems on the model of the
enzy me solutions to establish that low molecular sulfur
containing compounds with a free thiol group (cysteine,
glutathione) can restore the activity of en zy mes after their
post-radiation inactivation. The first ideas of the plausible role
of the capture by the protector of water radiolysis products and
the successive reduction of the indirect effect of radiation were
proposed by Dale et al. (1949).
According to the hypothesis proposed by Bacq and
Alexander (1955), sulfur containing radiat ion protection
agents can enter into competitive relations with water
radiolysis products by capturing them. They were the first to
suggest the possible involvement of rad ioprotectors into the
chemical processes of the reduction of radicals in
macro molecules through donating hydrogen by the thiol
group in the mo lecule of a pharmacological preparat ion
(Alexander & Charlesby, 1954). This idea is based on the
possibility of the absorbed radiation energy to migrate from a
macro molecule to the protector molecule through protonation
by the radioprotector of the macromolecule radical with its
subsequent chemical reparation. The importance of the ability
of SH-groups of the protector to form dis ulfide bonds with
proteins in the mechanism of the radiation protective action of
cysteamine was justified in the studies by Eldjarn and Pihl
(1956).
The significance of low mo lecular weight thiols in the
implementation of the “o xygen effect” was first shown in the
in vitro experiments on DNA and enzy mes by Hutchinson
(1961), as well as on the bacteriophages by Howard-Flanders
(1960). The ability of thiols to cause chemical reparation of
radicals of the Rtype by donating hydrogen from the SH-
group could ensure the participation o f sulfur -containing
compounds in the competitive process between the fixat ion
of the potential radiation damage by oxygen with the
formation of pero xides of the ROO type and their correct ion
by reducing equivalents in the cell.
Formation of the hidden potential lesions in
macro molecules after exposure to radiat ion was previously
 Journal of Radioprotection Research(2014) 15-36
justified by Eidus and Ganassi (1959). According to his
viewpoint, thiol groups and other reparative components of
the cell virtually do not restore the structure of DNA through
the mechanism of chemical reparation, but only modify the
damage in a favourable for further enzy mat ic reparat ion
direction. These processes act as competitive in relation to
the damaging action of oxygen in the environment.
The improvement of the effectiveness of a number of
radioprotectors from the group of aminothiols in the absence
in that period of time o f any significant progress in the
search for new mo re active pharmaceutical drugs could be
made in two directions. First, it was possible to use the
synergy in the action of the radioprotectors with different
mechanis ms for the imp lementation of radiat ion protective
properties, and second, to reduce the toxicity of aminothiols
to provide the possibility of using higher doses of the
pharmacological preparation.
It was found that the radiation protective effect of sulfur-
containing radioprotectors enhanced after their co mbined use
with pharmacological drugs that cause hypoxia, or und er
conditions of exogenous hypoxic hypo xia (Mayer & Patt,
1953; Devik, 1954). The greatest success in radiat ion
protection was achieved when using the pharmaceutical
composition that contains AET, cysteine, glutathione, and
serotonin (Jacobus, 1959, 1961; Maisin et al., 1968). In the
experiments on mice and dogs, the radiation protective
effectiveness of the pharmaceutical co mposition reached the
maximu m theoretically possible value with DRF equal to 2.8.
The comb ined application of th is pharmaceutical formulat ion
with the bone marro w t reat ment resulted in the DRF of 3.7
for radiation protective agents (Maisin et al., 1968).
In fact, further develop ment of the prevention of
radiation damage has led to the creation of a new special
branch of science – radiation pharmacology, whose subject
of study now is not only the search for new radioprotectors
or research into the mechanisms for chemical protection
against ionizing radiation, but also the improvement of the
estimation criteria for the effectiveness of radiation
protective agents, as well as peculiar features of
pharmacodynamics and pharmacokinetics of medicinal
preparations in the exposed organism. A great contribution to
the development of radiation pharmaco logy has been made
by professor Saksonov (1968, 1975). He was the first
researcher to find in the 1950s the possibility of reducing the
toxicity of aminothiols with the maintenance of their activ ity
by investigating the effect of d ifferent pharmaceutical d rugs
on the tolerability and radiation protective properties of
radioprotectors (Belay et al., 1960). The pharmacolog ical
compositions based on these principles allowed the reduction
of the toxicity and increase in the effectiveness of
aminothiols with the achievement of the optimal radiat ion
protective effect of rad ioprotectors on large animals (Vasin et
al., 1970a).
In the 1970s, the search for new effective radioprotectors
was completed with the discovery of the most active to date
radiation protective agents, which in the experiments on
17
large animals (dogs, monkeys) had a 100% radiat ion
protective effect after exposures to radiation at the doses
close to LD100 . This concerns, above all, the radioprotectors
WR-2721 (Davidson et al., 1980) and indralin (Ilyin et al.,
1994; Vasin et al., 1977, 1997).
The preparation WR-2721 (A kerfeldt et al., 1967;Piper et
al., 1969, 1975) was developed in the Walter Reed Army
Institute of Research, Washington, USA and represents itself
3-aminopropyl-2-aminoethylthiophosphate (amifostine). The
most comp lete pharmaco logical estimat ion of its radiation
protective properties was presented by Yuhas et al. (1969,
1970, 1971, 1973) and Harris and Phillips (1971).
Amifostine fro m the group of aminoalkylthiophosphates
release the residues phosphoric acids in the process of their
metabolism in tissues, and act as corresponding
aminoalkylthiols in the cells. An example is the rapid
reduction in blood of the radioprotector WR-2721 to the
active metabolite WR-1065 with a free th iol group, the
concentration of which is maintained in the equilibriu m state
with its disulfide form:
RS-PO3 HNa (WR-2721) → RSH (WR-1065)
→RSSR(WR-33278), where R = H2 N(CH2 )3 NH(CH2 )2 .
Clear advantages relating to the protective activity of the
radioprotectors from the group of aminothiols over
endogenous compounds at the cellular level are likely to be
related to their differences in the sign and magnitude of the
charge number of their mo lecules. The ro le of the amino
group in the co mposition of the aminothiol molecu le is
mu ltifunctional: it elevates the reactive properties of the thiol
group (pK (SH) = 7.7 – 8.6), thus providing a positive charge
of the radioprotector molecule, wh ich increases its
concentration on a negatively charged DNA mo lecule, as
well as allows the establishment of stronger chemical bonds
between amino disulfides and a macro mo lecule (through a
diamine group). The optimal distance between the mercapto
and amine groups in the aminothiol molecu le is the carbon
chain consisting of 2 - 3 carbon atoms. A ll effect ive radiat ion
protective substances from the groups of aminothiols and
aminothiophosphates have a positive charge number (for
cysteamine Z = +1, WR-2721 Z = +2), while endogenous
thiol co mpounds are neutral or negatively charged (for
glutathione Z = ─ 1 and for cysteine Z = 0).
The second factor, whose important role has recently
been addressed, is the changes in the quaternary structure of
DNA as a result of neutralization of its molecu le by
aminothiols man ifested in the spring-like shrinkage of the
mo lecule and contraction of the channels in its strand with
the disguise of the sites where the access of the hydroxyl
radical to the C4 desoxiribose of DNA is available (Savoye et
al., 1997).
The third factor, which may underlie the biochemical
basis for the positive effect of aminothiols on the enzymatic
stage of the implementation of the oxygen effect, is
associated with the ability of d isulfides of aminothiols to
form d iamine bonds with the DNA mo lecule with its fixat ion
in the liquid crystal structure (Jellu m, 1965; Bro wn, 1967).
18 Journal of Radioprotection Research(2014) 15-36
Since the charge nu mber of the molecu le doubles in the
disulfide form of aminothiols, the concentration of radio -
protectors in the form of disulfides on the surface of DNA
significantly increases. For examp le, condensation of the
disulfide WR-1065 (WR-33278) with Z = +4 on the DNA is
20 times higher than that in the solution (Newton et al.,
1996). One mo lecule of WR-33278 fixes four DNA
nucleotides (Savoye et al., 1997).
Taking into account that the charge number of the DNA
mo lecule is negative, a positive charge of aminothiols
predetermines their electrostatic attraction to the DNA
mo lecule and their condensation on the surface of the
macro molecule with the simu ltaneous repulsion of the
endogenous protective compound of glutathione (Vasilescu
et al., 1980; Zheng et al., 1992). According to the data by
Newton et al. (1996), this leads to the increase in the
concentration of the thiol form WR-2721 (WR-1065) on the
surface area of the DNA by 6 t imes. Recently, new family of
aminothiols has been investigated (Copp et al., 2011, 2013;
Peebles et al., 2012). A minothiol PrC-210 [3-(methylamino)-
2-((methylamino)methyl)propane-1-thiol] has as highest
protective effect as amifostine, but neither decrease arterial
pressure (AP) nor induce vomiting (Soref et al., 2012).
2. Realization of Oxygen Effect in Cells
and Organism
The “oxygen effect” as a radiobiological phenomenon
man ifests itself at the molecular, cellu lar and organism level
in practically all biological objects as the acceleration of the
radiation damage with the increased o xygen concentration in
the mediu m as compared with anoxic or hypo xic conditions.
The essence of the “o xygen effect” wh ich finds its
man ifestation at different levels of the biolog ical
organization in the wo rld of plants and animals lies in th e
increased radiation damage o f DNA, membranes, proteins,
carbohydrates in the presence of oxygen and transition
valence ions (Fe +2 , Cu + , Zn +2 ). Reactive o xygen species are
formed as a result of rad iation-chemical reactions:
superoxide anion radical (О2 ¯), hydropero xide rad ical
(НО2 ), atomic o xygen, o xygen in the long-lived excited
form, singlet oxygen (О2 ´).
These reactions proceed at a high speed, which is crit ical
for the reactions limited by the diffusion of molecu les.
Reactive o xygen species are unstable compounds. The
lifetime o f the superoxide anion radical is equal to 10 ms, of
the singlet oxygen - 2 ms, the hydroxy l radical НО - the
fractions of microseconds.
Radiat ion-chemical absorption of oxygen in the
environment after its exposure to radiation at a dose of 10 Gy
is close to 5 microM/L; under hypoxic conditions, the
reduced absorption of o xygen in these reactions is observed
together with the decrease of the o xygen concentration in the
environment <1 microM.
Organic co mpounds become activated and reactive to
physical and chemical interaction with o xygen during the
radiolysis. The activated state of the macro molecules lasts
for no more than milliseconds.
The damage of the macro molecu le is terminated with the
formation of pero xy l radicals: R + O2  ROO (К ~
109 •M –1 •c–1 )
Radio lysis products of organic compounds do not differ
in their chemical activity regardless of the direct or indirect
mechanis m of their damage during their interaction with
oxygen. Pero xides and epo xides of nitrogenous bas es are
formed in the DNA in the presence of o xygen, after that they
are ruptured, thus increasing the total amount of damage of
DNA bases by more than 3 times. The number of single-
stranded and double-stranded DNA breaks can increase 2 -
2.5 times under the influence of o xygen (Frey et al., 1974;
Ward, 1975).
The formation of reactive o xygen species constantly
occurs in the cell under physiological conditions without any
exposure to the damaging doses of ionizing radiat ion and is
the result of the inco mplete one-electron (fo rmation of О2 ¯),
two-electron (formation of H2 O2 ) or three-electron
(format ion of NO) reduction of O2 instead of its complete
four-electron reduction with the formation of water H2 O.
ROS
О2  О 2
¯
 Н 2О 2  ОН  Н 2О
Fe
2+
 Fe
3+
c a ta la s e
G S H -p e ro x id a s e
Fig. 1. The process of incomplete reduction of oxygen
Figure 1 shows the pattern of events aimed at the o xygen
reduction in the cells of the organism.
О2 ¯ can pass through memb ranes, diffuses in the cell
over long distances in the complex with Са +2 and Zn +2 . О2 ¯
is a reducing agent of Fe +3 in cytochrome C, an o xidizing
agent in the case of interaction with ascorbic acid, tocopherol,
and hydroquinone. When cells are exposed to the action of
interleikin-1 and tu mor necrosis factor О2 ¯, it plays a ro le of
the intracellu lar secondary mediator in the implementation of
their impact on cells.
Superoxide an ion radical О2 ¯is reduced to hydrogen
peroxide H2 O2 under the action of enzyme superoxide
dismutase (SOD) (McCord et al., 1978): H2 O2 is destroyed
by catalase and glutathione peroxidase. The damaging effect
of О2 ¯ is limited by superoxide dismutase (SOD). In the
case of the abundant format ion of О2 ¯ exceed ing the
possibilit ies of SOD, super o xide an ion radical init iates by
the Haber-Veiss reaction and in the presence in the
environment of Fe ++ and H2 O2 the formation of НО, the
most active oxidizing agent.
НО radicals can also arise without the participation of
О2 ¯ by the Fenton reaction through semiquinone forms in
the presence of transition valence metals (mo re often with
the ions of iron Fe +2 , sometimes Cu +).
 Journal of Radioprotection Research(2014) 15-36 19

The presence in the organism of system antio xidants Oxygen enhancement (OE) of the radiation damage in cells
cerruloplas min and transferin which can fix and be a place to at different levels of oxygen is reliably described by the
deposit the data about the metal ions in the bound inactive equation proposed byAlper and Howard-Flanders(1956): OE
form prevents the formation of hydro xyl rad ical НО or the = (m [O] + K)/([О] + К), where m is the maximu m
development of free rad ical react ions. The limit ing factor for multiplicity of the possible OE in the given conditions, K - the
the formation o f НО is the en zy mes such as catalase, oxygen content in the medium, at which ED50 of the OE effect
glutathione peroxidase that convert H2 O2 to H2 O. is observed, [O] is the oxygen concentration under study in the
In addition to the cellular antio xidant en zy mes: SOD, mediu m. The K value for the bacterial cells is close to 4 - 6
catalase and glutathione peroxidase, and system antio xidants: mcM of o xygen, for animal cells - 8 -10 microM O2 . The
cerruloplas min and transferin binding the abundant transition maximu m enhancement (m) of the radiation damage for
metals (Fe, Cu), the antiradical system also includes natural mammalian cells is close to 3.
antioxidants: ubiquinone, glutathione, ascorbic acid, α-
tocopherol, β-carotene, bioflavonoids, reduced nicotinamide
nucleotides. 3. The Role of Endogenous Sulfur-
Some co mponents of cellu lar respiration can also play a Containing Compounds in the Radi-
part of endogenous protectors. Cytochrome C and o ther
oxidation-reduction enzy mes are capable of utilizing osensitivity of Cells
hydrated electrons and hydrogen atoms.
Oxygen enhancement of the radiation damage of biological
The realization of the “o xygen effect” at the cellular level
objects is associated with the concentration of oxygen and
depends on (1) the o xygen tension in the tissues of the
thiol groups in the cell, as well as with the value of dose and
organism, and (2) the state of its antioxidant system. The
dose rate and LET of ionizing radiat ion. Oxygen does not
oxygen content in cells, in its turn, is defined by the
alter the radiosensitivity of cells at low radiation doses of
vascularization level and the state of microcircu lation in
0.25 – 1.25 Gy , though at high doses it enhances the
tissues, the velocity of the local blood flow, the permeability
radiation damage 2-3 times. The modifying effect of o xygen
of the cell memb ranes for oxygen and its solubility in
is reduced with the higher LET of ionizing radiat ion, when
subcellular structures of the cell, as well as by the intensity
the role of reparative processes is reduced at the molecular
of the oxygen consumption by the cell.
and cellular levels.
The highest vascularitzat ion and associated oxygenation
The importance of thiols in the radioresistance of cells
take place in the brain t issues, kidneys, liver, spleen and
was first shown Bott and Lundgren (1964) in the
bone marro w (40 - 60 microM O2 ). A lower o xygen tension
experiments on bacteria. The concentration of glutathione is
is observed in the skin, testes (20 microM), and intestinal
some orders of magnitude higher than the level of radiolysis
mucosa (7 - 10 microM) (Travis, 1984).
products in the cell after the exposure to lethal doses of
In the case of the reduction of the oxygen content in the
radiation. In th is situation, glutathione reduces the maximu m
environment up to 15-25 microM animal and human cells
oxygen enhancement of radiation damage of the cell ~ two-
experience a state of the moderate hypoxia, at 7 - 12 microM
fold. If we consider the situation in a living cell, then it
and <5 microM – an expressed and deep hypoxia,
contains 0.5 - 2 mM of reduced glutathione and an average
correspondingly. The part of the NADH respiratory chain -
of 15 microM of o xygen. After irradiat ion at a dose of 10 Gy,
KoQ is the locus most sensitive to the lack of oxygen where
0.5 - 5 • 10-2 microM of radiolysis products of water are
there may occur difficulties in transferring electrons when the
formed (main ly rad ical НО and е), wh ile changes of the
oxygen concentration drops lower than <30 microM. Critical
DNA bases occur ~ twice less often.
levels of the oxygen concentration in the medium for the
The natural level of o xygen in the cell represents a
oxygen dependent energy processes are 10 - 20 microM. The
serious competition to glutathione for the interaction with the
reduction of the ATP synthesis is observed at the oxygen
DNA radicals after radiat ion, thus reducing the protective
concentration of <20 microM.
role of thiols more than twice on condition of their normal
ED50 of hypoxia by the shift of the NADH / NAD +
content in a cell. The deficiency of g lutathione in the cell
relation is equal to 12.6 microM, by the change of the ATP /
sufficiently limits the manifestation of the o xygen effect
ADP relation - 7 microM, KmO2 for the respiratory chain of
(Morse & Dahl, 1978; Edgren et al., 1980; Midander et al.,
cells is 1.9 microM of oxygen (Jones et al., 1978). There is the
1982).
oxygen intracellular concentration gradient. Provided the
Another endogenous source of free SH-groups in the
mediu m contains 6 microM of oxygen, then the cytoplasm has
cells is metallothioneins, which represent themselves low
2 - 5 microM, mitochondrial clusters <2 microM of oxygen.
mo lecular proteins capable of binding heavy metal ions
The dependence of the radiosensitivity of mammalian cells
(copper, zinc, cad miun, etc.) and containing up to 30% of
in vitro on the oxygen tension in culture was first described by
cysteine. Both glutathione and metallothioneins exogenously
Gray et al. (1953). The remarkable manifestation of the oxygen
introduced into the organism have a rad iation protective
effect is observed when the oxygen tension in the medium is
effect (Koterov et al., 1995).
above 40 microM.
20 Journal of Radioprotection Research(2014) 15-36

There are several possible mechanis ms for participation As for the cell nucleus, whose radiation damage is a
of low mo lecular weight SH-co mpounds in the protective determining factor for the viability of the cell, the reacti on
effect when liv ing cells are exposed to the action of ionizing of type I (Figure 2), that is the interception of water radicals
radiation: (1) the interception of water radicals, (2) the by natural protective compounds in the environment, is not
migrat ion of energy or charge fro m macro mo lecules to SH- determining because of the significantly higher level of the
compounds by means of the proton transfer fro m SH- DNA reactivity relatively to the hydro xyl rad ical НО (К ~
compounds to the radical of the macro mo lecule followed by 1012 М –1 с–1 ) than that in the endogenous thiol compounds (К
the chemical reparation of its init ial state, and (3) the ~ 1010 М –1 с–1 ).
formation of mixed disulfides. After the formation of DNA radicals following the direct or
indirect exposure to radiation, competitive reactions of type II
Т  + S H  Т Н + S  - c h e m ic a l re p a ra tio n  s u rv iv a l o f c e lls
for the interaction with them of endogenous thiol compounds
III and oxygen are most likely. The mechanism of the “oxygen
SH
effect” finds its realization in competitive relations between
I II molecular o xygen and SH-groups of endogenous compounds
SH SH for the interaction with macromolecular radicals (Ho ward-
e n z y m a tic re p a ra tio n  s u rv iv a l o f c e lls
Flanders, 1960; Hutchinson, 1961).
Reactions of type III are possible between free radicals of
Т + ОН  Т + О 2  Т О О   c e ll d e a th the protein, glutathione, oxygen and DNA macromolecule. The
portion of extra DNA lesions due to this mechanism will be
small. The reactivity of the secondary radicals has the
p h a rm a c o lo g ic a l h y p o x ia  s u rv iv a l o f c e lls following order: DNA radical > protein radical > О2 >
glutathione radical. The influence of the protein radical on
DNA results in the formation of the slowly reparable DNA -
Fig. 2. Scheme for realization of the oxygen effect at the
protein crosslinks. The reaction proceeds at high concentrations
level of cell
of protein in the absence of oxygen. Glutathione is a trap for all
Figure 2 shows the main crit ical links in the the types of radicals
man ifestation of the oxygen effect at the cellular level and Reactions of type IY procede with the participation of
where the realizat ion of the protective action of endogenous peroxide free radicals and DNA pero xides. Nine mo lecules
SH-compounds is possible. of o xygen are absorbed in the DNA per 100 eV of the
The value of each of the above noted mechanis ms for the absorbed radiation energy. The reactivity of pero xide free
protective effect of low mo lecular weight SH-co mpounds radicals of DNA is sufficiently lower than that of DNA
can vary significantly depending on the predominance of the radicals. DNA pero xides are involved in DNA
direct or indirect mechanis m of the radiation damage for intramolecular reactions leading to the breaks of DNA bases
different sub-cellular structures. and ribose phosphate bonds. When DNA peroxides interact
As a result of co mpart mentalization of low mo lecular with hydrogen donors, they can be converted to hydro
weight substances in the cell by intracellular hydrophobic peroxides which are unstable and break with the formation of
memb ranes, the differences in their concentration in free radicals НО and R=O. The possibilities for the
individual structures can differ by 1 - 2 orders. Thus, there is chemical reparation of DNA pero xide co mpounds are
some heterogeneity both in the radiation damage of extremely limited and are not co mpetit ive with the system of
individual cell structures and in the value of the radiation enzymatic DNA reparation.
damage of part icular structures for the post-radiation cell The important final phase in the scheme for the “o xygen
death. effect” (Figure. 2) is the en zy matic reparation of damaged
The chemical co mposition of the cell nucleus includes macro molecules (Holmg ren, 1979; Eidus & Korystov, 1980).
DNA (0.5 mM, 1%), histone and non-histone proteins (10 A modified DNA frag ment can be removed only enzy mat i-
mM , 20%), RNA (1%), lipids of the nuclear matrix and cally through the mechanis m of the excision reparat ion of the
memb ranes (2 - 3%). Low mo lecular weight compounds of damaged DNA strand. The radiation damage of DNA indu c-
the nucleus are largely represented by reduced glutathione es an increased functional activity of the tumo r suppressor
(1-5mm). protein p53, wh ich part icipates in the intensified generation
The effect of o xygen sensitizat ion main ly leads to the of reactive o xygen species (ROS). ROS have the signaling
reduction of the endogenous defense of DNA by glutathione function in the activation of the two different ways of intra-
and proteins. Suppression of the format ion of DNA cellu lar regulatory cascades associated with kinase regulated
superoxide is more remarkable under the influence of by extracellular signals (ERK ½) and с-Jun N - terminal ki-
glutathione than high molecular proteins. The radiation nase (JNK). The ERK-cascade leads to the activation of the
protective effect of glutathione is possible at concentrations transcription nuclear factor kB (NF-kB), which causes the
of about 1 mM in the presence of o xygen, and 0.1 mM in the state of growth / differentiat ion in the cell, while JNK-
hypoxic conditions. cascade initiates apoptosis.
 Journal of Radioprotection Research(2014) 15-36
The increase of the intracellular level of g lutathione in-
hibits the process of post-radiation apoptosis. The state of the
redox system of cells predetermines the level of their radio-
sensitivity. As previously stated by Eidus and Korystov
(1980), thiol groups and other reduction components of the
cell mod ify the damage to the direction which is favorable
for the further en zy matic reparat ion. These processes act as
competitive in relation to the damaging effect of oxygen in
the medium.
The potential protective possibilities of the above reac-
tion lie within the limits of the rad ioprotective effect of re-
duced glutathione close to the DRF = 1.2, wh ich can range
fro m 20% up to one third of the observed radiation protec-
tive effect in aminothiols.
4. The Modification of Radiation Dam-
ages on Physicochemical and Bio-
chemical Levels by Radioprotectors
Current ideas about the mechanism for the radiation protec-
tive action of radioprotectors have evolved historically as the
accumulat ion of the hypothetical notions about very i m-
portant scientific facts relating to the action of these co m-
pounds at different levels of the imp lementation of the radio-
biological effect. However, all o f the proposed hypotheses
explaining the radiation protective effect of pharmacolog ical
compounds, unfortunately, fail to provide a uniform and
generalized theoretical approach to the problem concerning
the mechanism for the action of radiation protectors as a
whole. The co mplexity of the problem is connected with the
possibility of the simu ltaneous implementation of several
mechanis ms for the radiat ion protective action (Bacq & A l-
exander, 1961). Nevertheless, it is important fro m a scientific
and practical point of view that the level of current theoreti-
cal develop ments allows quite a correct approach to the as-
sessment of the ro le of indiv idual plausible units, as well as
the sequence of stages in the imp lementation of the radiat ion
protective properties of different groups of radiation protec-
tive compounds.
The mechanism for the action of radioprotectors is man i-
fested either in the pharmacological reduction of the oxygen
content in the cell or in the direct participation of thiol
groups in the molecu les of sulfur containing radioprotectors
in co mpetitive reactions with o xygen for the products of ra-
diolysis of vitally impo rtant macro molecules of the cell (Ei-
dus & Korystov, 1980).
According to the hypothesis proposed by Bacq and Ale x-
ander (1955), sulfur containing rad iation protective agents
can enter into competitive relations with the products of wa-
ter radio lysis, capturing them and thus preventing the damag-
ing action of pero xide co mpounds on macro mo lecules of
cells. They were the first to suggest the possible involvement
of radioprotectors in the chemical processes of the reduction
of radicals in macro mo lecules by donating hydrogen from
the thiol group of the molecule of the pharmaceutical prep a-
21
ration (Alexander & Charlesby, 1954).
The significance of low mo lecular weight thiols for the
implementation of the “oxygen effect” was first shown in in
vitro experiments on the DNA and enzy mes by Hutchinson
(1961), as well as on the bacteriophage by Howard-Flanders
(1960). The ability of thiols to cause chemical reparation of
radicals of the R type by donating hydrogen fro m the SH-
group could ensure the participation of sulfur containing
compounds in the competitive process between the fixat ion
of the potential radiat ion damage by o xygen with the fo r-
mat ion of pero xides of the ROO type and their correction
by the reducing equivalents of the cells.
Recent research into the possible ro le of the direct
chemical interaction of sulfur-containing radioprotectors and
a DNA mo lecule indicates that they can play a key role in
the imp lementation of a high radioprotective efficiency
which is inherent to them.
The general scheme for the possible interference of
sulfur-containing radioprotectors at the level of the primary
radiation-chemical and biochemical processes in the cell and
the mechanis m of realization of their radiat ion protective
effect is analogous to that proposed above for endogenous
thiol compounds (Figure 2).
When a concentration of the radioprotector in the DNA
cell is 0.5 mM, it can have the radioprotective effect on
macro molecules in co mpetition with the active radiolysis
products of water at a concentration of no lower than 30 mM.
Dimethyl sulfo xide has a radiation protective effect at such
high concentrations (65 mM) due to the direct interaction
with the hydroxyl radical.
The additional protection of DNA due to the inactivation
of radiolysis products of water by other known effective
radioprotectors is insignificant (it does not exceed 20% at
their actual concentrations in the cell) because the doses
transferred by most of them do not exceed 3 mM . The basic
inactivation processes of radiolysis products of water in the
cell are associated with their reactions with proteins and
endogenous glutathione, whose concentrations reach 5-10
mM (Sapezhinskiy et al., 1992)
The main role in the realization of radioprotective
properties of the known most active sulfur-containing
radioprotectors at the level of radiation-chemical reactions is
assigned to their ability to interact directly with the radiolysis
products of macro molecules, thus causing their reparat ion
(reaction of type II in Figure 2), rather than to the
competitive relationships with macro molecu les for the
radiolysis products of water.
The protective effect of sulfur-containing radioprotectors
is associated with a free SH-group in the co mposition of
their mo lecule. The possibility to transfer the hydrogen atom
fro m the SH-group of the protector to the rad ical of the
macromolecules provides its chemical reparation:
М + HS–R  MH + R–S (Alexander & Charlesby, 1954).
Chemical reparat ion of the radica l of deo xyribose in
DNA by a hydroxyl radical under dehydrogenation of the
22 Journal of Radioprotection Research(2014) 15-36
mo lecule is of crucial importance for the reduction of the
frequency of chromosomal aberrat ions in the cells under the
influence of radioprotectors (Fahey et al., 1988; Held et al.,
1988): Deoxyribose (-H) • + RSH = deoxyribose + RS •
The rate constants for the transfer of a hydrogen atom
fro m cysteamine to the radical of a macro molecule are close
to 106 – 4•108 М –1 •с–1 (Оrmerod et al., 1967). The known
sulfur-containing radioprotectors in the reactions of
interaction with the radicals of macro molecu les and DNA
reach the rad iation protective effect at the concentrations ~ 2
- 3 t imes lower than it is possible under the influence of
endogenous thiols.
However, the interaction of aminothiols with the DNA
mo lecule can be more specific, than a simple adsorption on
the macro mo lecule. Jellu m (1965) showed the possibility of
forming a chemical comp lex between cystamine and DNA
through a diamine bond. According to his data, this co mplex
had a greater thermal stability than unprotected DNA
mo lecules. Also considered was a hypothesis (Jellu m, 1965;
Bro wn, 1967) about the ro le of stabilizing the DNA helix
regions which are not wrapped with h istones through a
diamine bond by using radioprotectors in the improvement
of its resistance to post-radiation degradation and
man ifestation in this way of rad iation protective
effectiveness of cystamine.
The role o f the increased level of endogenous thiols in
the realizat ion of the radiation protective effect of
aminothiols was first substantiated in the hypothesis by
Graevsky (1967). According to the viewpoint previously
expressed by Eidus and Korystov(1980), thiol groups and
other reparative components of the cell actually do not
restore the structure of DNA through the mechanis m of
chemical reparation; they only modify the damage in the
direction favorable for further enzy matic reparation. These
processes act as competitive in relation to the damaging
effects of oxygen in the environment.
This phenomenon is mainly associated with the positive
charge of aminothiols due to the presence in a molecule of
the amino group, which should be located no further than 2 -
3 carbon atoms fro m the free thiol group of the
radioprotector to provide the optimu m effect of the particu lar
chemical structure. Due to the positive charge of their
mo lecule and by means of electrostatic attraction aminothiols
can closely interact with the DNA mo lecule bearing a
negative charge (Zheng. et al., 1988, 1992; Spotheim-
Maurizot et al., 1991).
However, the interaction of aminothiols with the DNA
mo lecule can have a more specific character than a simple
adsorption on a macro mo lecule. Jellu m (1965) was the first
to show the possibility of forming a chemical co mp lex
between cysteamine and DNA through a diamine lin k. Due
to a high positive charge of the disulfide WR-1065 (Z = +4)
on the surface of DNA the amount of the radioprotector
fixed through a diamine bond is 3 times higher than by
means of its electrostatic adsorption (Newton et al., 1996).
Neutralization of the negative charge of DNA during the
formation of a new co mplex of disulfide WR-1065 with
DNA leads to the reduction of the DNA molecu le along the
axis with the formation of a liquid crystal spring -like
structure with the narrowing of the channels between the
DNA helices (Savoye et al., 1997). The latter mechanism can
be effective for the reduction of the radiat ion damage to
DNA due to the decrease in the number of the sensitive
places to be attacked by the hydroxyl radical at a density of
fixation of mo re than one mo lecule of the protector per 4
nucleotides of DNA (Savoye et al., 1997).
The second way to implement radiation protective
properties of aminothiols is associated with their ability to
raise the level of reduced equivalents in the cells and, in the
first place, endogenous thiols: in terms of the level of
glutathione and cysteine - fro m 2-3 up to 10 times (Ward man
et al., 1992). Moreover, this process takes place without the
involvement of the intracellular synthesis of glutathione de
novo, it is rather due to the increase of the transport of
cystine into the cell (Jokay et al., 1998). A miothiols inhubit
catalase (Lesort et al., 2003) and stimu late Mn-SOD activ ity
(Murley et al., 2006, 2007). The redo x potential of thiol
compounds is an important determinant for the manifestation
of their activity (Grdina et al., 1995).
The elevated level of endogenous thiols under the
influence of aminothiols causes the development in the cell
of the “recovery stress” or “biochemica l shock” (according
to the terminology of Z. Bacq), which, in particular,
man ifests itself as the swelling of mitochondria in the cells
(Lehninger & Schneider, 1959; Neubert & Lehninger, 1962).
One of the constituents of this reaction is the activation of
the p53 tumo r suppressor under the action of aminothiols
(Pluquet et al., 2003b). Th is reaction develops given the
interaction of ext racellular thiols with the cell membrane.
The transfer of the stress signal to p53 is carried out through
the c-Jun N-terminal kinase (Pluquet et al., 2003a).
It is likely that the conformational structure of DNA
undergoes some changes under the influence of aminothiols
associated with the condensation and stabilization of
chromatin, thus enhancing the interplay between the
transcriptional factors (nuclear factor - kappaB, protein-1
activator and tumor supressor p53) and DNA, which leads to
the subsequent activation of a number of genes. In addition,
aminothiols due to their ability to block topoisomerase II, as
well as under activation of p53 through the inhibitor of
cycline-dependent kinases p21waf-1 prevent the passage of
cells through the cell cycle in the G1 phase and provide more
opportunities for the reparation of the DNA radiat ion
damage (No rth et al., 2000). Th is effect underlies the
antimutagen effect of the radioprotector.
A characteristic pharmacodynamic property of sulfur
containing radioprotectors is almost a d irect correlat ion
between the level of exp ression of radiat ion protective
properties of aminothiols in terms of DRF and the
administered dose of the pharmaceutical preparation and its
concentration in the radiation-sensitive tissues (Patt et al.,
1953;Doherty, 1960;Hasegawa et al., 1970; Vasin et al.,
 Journal of Radioprotection Research(2014) 15-36
1970b) (Figure. 3).
The effect of “saturation” observed by a number of
authors (Doherty, 1960) in the studies when high doses of
aminothiols are ad min istered is eliminated to a certain extent
by reducing the toxicity of the pharmaceutical drug
(Saksonov, 1975). In fact, the main restriction in the
man ifestation of radiation protective activity of aminothiols
in hu mans up to a theoretically possible value is a poor
tolerability of the pharmaceutical drug, side and to xic effects
of the radioprotector.
Limitations in the manifestation of radiation protective
properties of radioprotectors with the increase in the
radiation dose are associated with their lower effect iveness
in the case of the intestinal syndrome and a practically
complete absence of the action under exposures to radiation
of CNS as co mpared with their high act ivity aimed at the
protection ofthe hematopoietic system in humans (Yuhas et
al., 1969).
Radioprotectors are of great practical interest since they
have the ability to rapid ly (within some minutes) increase the
resistance of the organism to radiation exposure and possess
high radiation protective properties, including during
exposures to radiation at super lethal doses (up to 10 - 15
Gy ), which are absent in other groups of radiation protective
agents.
An important common characteristic feature of radiat ion
protective agents is the value of the reduced severity of
radiation damage after their ad ministration, which is
theoretically limited to a certain threshold associated with
the possible modifications of the radiosensitivity of cells and
the whole body of animals and humans.
Radiat ion protective effect of radioprotectors can not
exceed in its intensity the radiosensibilising action of o xygen
dissolved in tissues, which can increase the radiat ion damage
to cells by 2 - 3 times.
Since the direction of the action of a radioprotector is
competitive relating to the radiobio logical effect of ionizing
radiation, the radiat ion protective activ ity of substances can
be expressed as a dose reduction factor (DRF). At present,
the dose modificat ion factor (DMF) is a more widely
accepted term, wh ich represents the ratio of the average
lethal dose (LD50 ) of rad iation in the experimental group
(with administration of the protective agents) to LD 50 of
radiation in the radiat ion control group. This index is the
most informat ive for a co mparative quantitative assessment
of the potentials of a radiation protective preparation,
because it, according to Ariens et al.(1957), to a certain
extent reflects the limits of manifestation of its “intrinsic
activity”.
Given the whole variety of plausible interpretations for a
particular participation of a radioprotector in the format ion
of a high radioresistance of the cell, the solution to the
fundamental question about the predominant mechanis m of
their action mainly depends on the availability of clear
quantitative estimates of any modifications in the cell, as
well as their interplay with the dose-dependent nature of the
23
pharmacological effect of a radioprotector.
When making a pharmacological analysis of the
protective effect of radioprotectors, it is noteworthy that the
substance should be present in the bio -object at the t ime of
irradiation to manifest its radiation p rotective effect, thus
indicating the importance of the direct participation of the
drug in the primary radiochemical reactions. Nonetheless, it
is important fro m the scientific and p ractical viewpoint that
the level of the current theoretical developments makes it
possible to provide quite a correct approach to the
assessment of the role of the particular possible links , the
sequence of stages in the realizat ion of the radiat ion
protective properties of various groups of radiat ion
protective compounds.
Given the whole variety of plausible interpretations for a
particular participation of a radioprotector in the format ion
of a high radioresistance of the cell, the solution to the
fundamental question about the predominant mechanis m of
their action mainly depends on the availability of clear
quantitative estimates of any modifications in the cell, as
well as their interplay with the dose-dependent nature of the
pharmacological effect of a radioprotector.
5. The Role of Pharmacological Hypox-
ia in Tissues in the Mechanism of the
Action of Radioprotectorsincluding
that from the Family of Aminothiols
The first informat ion about the ability of biogenic amines
having a pronounced radiation protective effect to reduce the
oxygen content in the hematopoietic tissues on the examp le
of serotonin allowed van der Meer and van Bekku m (1959,
1961) to propose a hypothesis about the role of
pharmacological hypoxia as the main mechanism of the
radiation protective act ion of these compounds. The studies
of van den Bren k et al. (1959, 1961, 1962) were the first to
show the possibility to eliminate or substantially attenuate
the radiation protective effect of adrenaline, histamine,
serotonin, and aminothiols under the influence of the
increased oxygen pressure.
The role of pharmaco logically induced disturbances of
the local b lood flo w in hemopoietic tissues in the
implementation of radiation protective action for many
derivatives of serotonin at different doses and methods of
application was confirmed later (Zherebchenko&Suvorov,
1963; Hasegawa et al., 1967; Iarmonenko et al., 1970; Vasin
et al.,1979, 1984, 1987, 1997, 1999).
The real opportunities for the modification of the
radiosensitivity of the organis m in the case of
pharmacologically induced disorders of the oxygen delivery
to tissues can be comprehended based on the radiation
protective effect of the hypoxic hypoxia in the experiments
on animals under conditions of the o xygen delivery to the
organism and its tension in the blood as low as twice or three
times. Table 2 shows the data available in the literature about
24 Journal of Radioprotection Research(2014) 15-36
the radioprotective effectiveness of normobaric hypoxic
hypoxia with respiration of gas hypoxic mixtures (GHM)
with the deficient o xygen content from 15% to 5%. The first
evidence in favor of the increased resistance of animals to
irradiation during respiration of GHM with the o xygen
content of 10, 7 and 5% was obtained by Dowdy et al. in
1950.
The effect of GHM in rats is lower than that in mice. In
the experiments on large animals (dogs) the radiation
protective action of GHM was even less pronounced and did
not exceed 50-60%, which is not higher than 1.2-1.3 in terms
of DRF (Hasegawa et al., 1967; Strelkov et al., 1974; Vasin,
1986, 1997).
The level of cell hypo xitension with the reduction of the
oxygen delivery to tissues is, in all probability, mainly
determined by the intensity of the tissue respiration, which,
for examp le, in dogs and rats is 5 and 2 times as low as that
in mice, wh ich causes the reduction of the radiation
protective action of GHM in these animal species (Vasin et
al., 1986, 1997).
Reduction of the radiation protective properties of
mexamine, an active derivative of serotonin, in the
experiments on rats and dogs, as compared with the effect on
small rodents is associated, as in the case of application of
hypoxic mixtures, with the influence of a lo wer in itial level
of the tissue respiration wh ich in many ways predetermines
the degree of a real decrease of pO2 in the cell (Vasin et al.,
1997).
The role of pharmacological stimu lation of cellu lar
respiration as a factor capable of inducing a significant
radiobiological t issue (bio-energetic) hypoxia was
demonstrated by the achievement of the 100% radiat ion
protective effect during the application of dinitrophenol, a
disconnector of the o xidative phosphorylation (Vacek et al.,
1964).
A high radioprotective efficiency of indralin in the large
animals (dogs, monkeys) is largely predetermined, in
addition to the vasoconstrictive effect, by its ability as an 1 -
adrenomimetic to stimu late the oxygen consumption in
tissues (Vasin et al., 1997).
Hypoxia in radiation-sensitive tissues can be also induced
by the blood circulation problems due to the pronounced
vasodilation, wh ich occurs under the influence of
radioprotectors of a number of aminothiols (Yuhas et al.,
1973). Studies by Allalunis -Turner et al. (1989, 1990) of the
estimation of the oxygen content in bone marro w cells o f
mice in v ivo have shown that following the application of
aminothiols, the hypoxic factor alone predetermines about
half of the observed radiation protective effects of amifostin.
A certain role in the develop ment of cellu lar hypoxia can
belong to the effect of aminothiols on the bioenergetics of
cells accompanied by their direct action on mitochondria
resulted in their swelling, the disturbances in o xidat ive
phosphorylation and the intensification of cellular respirat ion
(Lelievre et al., 1965; Firket et al., 1966; Skrede, 1966;
Vladimirov et al., 1970).
Finally, aminothiols in the course of their o xidation in the
cell are capable of reducing the intracellu lar content of
oxygen (Purdie et al., 1983). This can contribute to cellu lar
hypoxemia when aminothiols prevent the transport of
oxygen to the cell (Vachon et al., 1987).
The “o xygen effect” is taken into account in the pract ical
med icine when developing the ways of increasing the
radioresistance of the organism. Radiotherapy of cancer
patients currently utilizes hypoxic mixtures with the twice
lower o xygen content (GHM-10), which makes it possible to
reduce the radiation damage of the skin in the area of the
tumor project ion by 1 - 2 times. The most active
radioprotectors applied by man are known to have DMF
equal to 1.2 - 1.5. For so me tu mors, the application of
hyperbaric oxygenation during the therapeutic exposure of
patients makes it possible to increase to the same extent their
sensitivity to the action of ionizing radiation. The
phenomenon of the “oxygen effect” is also the basis for the
application of a nu mber o f rad io sensitizers fro m a series of
electron acceptor compounds.
Theoretically, the obstacle to prolongation of their
specific action is that the implementation of the radiat ion
protective effect at the cellular level is inevitably
accompanied by the development of drastic modifications in
its functional state, which is described in the radiobiological
literature as a “biochemical” shock (Bacqet al. , 1968).
Earlier, this phenomenon was discovered by Nasonov and
Aleksandrov (1940) as a nonspecific response of the cell to
the damaging action, which they named as “paranecrosis”.
Such state as this can only be maintained in warm-
blooded animals at normal body temperature for a certain
time period and results either in the develop ment of adaptive
reactions expanding the possibilities for the vital activ ity of a
cell in the new environ ment or in the irreversible shifts
leading to its death. According to the hypothesis proposed by
Z. Bacq, the development of “biochemical” shock is a
necessary condition for the imp lementation of the radiation
protective effect of a radioprotector. This is the reason for
why the termination of the adaptive processes in cells in
response to the action of radioprotectors and the withdrawal
fro m the state of “biochemical” shock should lead to the
reduction and cessation of the radiation protective action.
Another option is the death of cells fro m the to xic act ion of a
radiation protective agent. Thus, the duration of action of the
radioprotector is limited by the possible persistence of
viability of the cell under conditions of the severe restriction
to its metabolism.
Irrespective of all the variety of p lausible interpretations
of a particular participation of a radioprotector in the
formation of an enhanced radioresistance of the cell, the
answer to the principle question about the predominant
mechanis m of their action main ly depends on the definition
of the accurate quantitative estimates of any changes in the
cell and their relationships with the dose dependence of the
pharmacological effect of a radiation protective agent.
 Journal of Radioprotection Research(2014) 15-36
6. Pharmacological Properties of Ami-
nothiols
6.1. Pharmacokinetic of aminothiols
A rapid man ifestation of radiation protective properties of
aminothiols (within several minutes) after intravenous drug
administration is directly associated with the delivery of
radiation protective substances directly into the cells, first of
all, of radiosensitive tissues and achievement in them of the
maximu m concentration of the drug over a short period of
time (Eldjarn et al., 1956; Vladimirov, 1968; Titov, 1974;
Washburn et al., 1974; Utley et al., 1984).
The important peculiar feature of the pharmacokinetics of
aminothiols is their predominant accumu lation in
radiosensitive tissues, where their concentration is several
times higher than in the muscle tissue or blood of animals
Different pharmaceutical preparations of this group
(cystamine, A ET, cystafos, gammafos) have some
differences; however, their co mmon fundamental peculiar
feature is their predominant distribution in rad iosensitive
tissues. A comparative analysis of pharmacokinetics of
cystamine and amifostine given at equal doses shows that the
latter is less accumulated in the brain, wh ich is important for
its better tolerability by the organism as compared with
cystamine (Vladimirov et al., 1993; Utley et al., 1984). It
should be noted that aminothiols are mo re read ily
accumulated in the tissues with a higher level of
physiological regeneration. An act ive and selective delivery
of aminothiols into radiation sensitive tissues, in the liver
and kidneys, is most likely to be associated with the crucial
role of free SH-groups of endogenous thiols, especially
glutathione, in the maintenance of the cellular antio xidant
system involv ing glutathione (glutathione pero xidase and
glutathione reductase) in the tissues with a h igh metabolic
rate. The level of reduced glutathione in the liver and
kidneys is many t imes higher that its concentration in
muscles.
For example, a similar distribution in the tissue was
observed for 35 S-methionine, wh ich is not a radioprotector.
After exposure to radiation, the accumulation of 35 S-
cysteamine in t issues, except for the intestines, is reduced,
although the main outline of a characteristic pattern of its
distribution remains the same (Arbuzov et al., 1961).
The o xidation of sulfur-containing rad iation protectors, in
particular a free SH-group of aminothiols, which is
associated with the manifestation of radiation protective
properties of these compounds, occurs quite readily, and can
proceed non-enzymatically through free o xidation processes
in the presence in the med iu m o f o xygen or under the
influence of cellular oxygenases.
The duration of radiation protective action o f aminothiols
is maintained by high concentrations of the drug in
radiosensitive cells. This makes it clear why there is a direct
relationship between the expression and duration of the
radiation protective effect and the dose of aminothiols. In
humans treated with cystamine taken at a dose of 1.2 g, this
25
radioprotector is detected in the urine at the maximu m
concentration 1 hour after ad min istration in native or act ive
forms with free SH-groups, is secreted intensively for 2 - 3
hours and completely eliminated within 5 hours.
Amifostine (ethyol, gammafos) is dephosphorylated
under the action of alkaline phosphatase in the vascular
endotheliu m to the active metabolite WR-1065 with a free
thiol group, whose concentration is maintained in the state of
equilibriu m with its dis ulfide fo rm (Calabro-Jones et al.,
1985): RS-PO3 H2 (amifostine, W R-2721) → RSH (W R-
1065) ↔ RSSR
T1/ 2 of the native drug in the blood after its intravenous
infusion ad min istration for 15 minutes is 0.8 hour.
Amifostine is detected in the active form in the blood plasma
during 24 hours. T1/2 of WR - 1065 is 7.3 hours, T 1/2 of the
drug in the disulfide form is 8.4-13.4 hours (Korst et al.,
1997).
The drug penetrates into the cells of tissues in the
disulfide form through the polyamine transport system
(Newton et al., 1996). The concentration of the drug
accumulat ion in cells is 1.5 - 2 times higher than that in the
intercellular space.
The process of excretion of the degradation products of
aminothiols main ly occurs during the first 24 hours. Up to
one-third of the decomposition products of cysteamine
labeled with 35 S is excreted during 6 hours. The removal of
the degradation products of radioprotectors is delayed after
the exposure of the organism to radiation; however, their
composition undergoes no qualitative changes. The products
of the aminothiol transformat ion are excreted in the urine
(80%) and b ile (7%) main ly in the form of free sulfate and
taurine, and in s maller quantities in the form of hypotaurine
and cysteamine disulfoxide.
6.2. Influence of aminothiols on the process of cellular
respiration
Pharmacological effects of aminothiols at the cellu lar level
are directly related to the mechanis m for imp lementation of
radiation protective properties of radioprotectors. It is
accepted among radiation oncologists to define a nu mber of
cytophysiological and biochemical effects of aminothiols
which lead to significant changes in the functional state of
the cell as “biochemical shock” according to the hypothesis
proposed by Z. Bacq et al. (1968).
The first fact to which Bacq paid his attention was a
reaction of mitochondrial structures to the presence in the
med iu m o r aminothiols or reduced glutathione in the form of
a reversible swelling and changes in the structure of their
crysts (Lehninger et al., 1959; Firket, 1966). The swelling of
mitochondria is direct ly regulated by the state of the thiol-
disulfide equilibrium in the respiration chain.
Later, it was found that under usual conditions,
mitochondria normally undergo successively alternating
phases of swelling and reduction. Swelling is acco mpanied
by the release of energy (conversion of the chemical energy
of the electron transport through the membrane into the
26 Journal of Radioprotection Research(2014) 15-36
mechanical energy) and is stimu lated by calciu m ions, an
excess of oxidation substrate (succinate), thyroxine and other
hormones. Mitochondria shrink and restore their orig inal
structure under the effect of catalase, g lutathione peroxidase,
ADP, ATP and Mg 2+ ions. Under conditions of the
mitochondrial swelling, the nu mber of SH-groups increases,
while their amount drastically drops when mitochondria
shrink. An abundant reaction of the mitochondrial swelling
can lead to a destruction of the mitochondrial structure.
The changes in the structure of mitochondria in the form
of swelling are directly reflected in the operation of the
phosphorylation respiratory chain, namely , in the degree of
relationship between the processes of electron transport and
oxidative phosphorylation, wh ich is measured by the P/O
ratio. The maximu m values of the P/O ratio are observed
when the integrity of mitochondria is the highest. When
mitochondria swell, the d istance between the outer and inner
memb rane decreases and the flow of electrons through the
memb ranes accompanied by the release of energy in the
form of heat due to the intensification of the processes of
free oxidation increases.
The reduced level of the P/ O relat ionship represents itself
one of the ways of regulating the respiratory chain. A mild
and moderate uncoupling of oxidative phosphorylation is a
necessary condition for the accelerat ion of o xidation when
there is a physiological requirement for increased energy. A
high rate of the energy storage by the respiratory chain under
these conditions has a more b iological significance than the
optimu m effect iveness from the use of the oxidation
processes.
Thiol groups are involved both in the coupled and free
oxidation, and serve the basis for communication between
the two processes. Aminothiols elevated tissue respiration in
the experiments on mouse liver slices (Iarmonenko, 1961),
on the human, dog and pigeon erythrocytes (Kuznetsov et al.,
1964), and on the brain t issue homogenate (Kandasamy et al.,
1988)
Aminothiols have an effect on separation of respiration
and phosphorylation in cells, wh ich is acco mpanied by
mitochondrial swelling and intensification of free o xidation
processes (Lelievre et al., 1965; Firket et al., 1966; Skrede et
al., 1966; Vladimirov et al., 1970). A mifostine when added
to the mediu m was found to increase the temperature of the
tissue culture in vitro (Purdie et al., 1983).
The ad min istration of cystamine in hu mans at a dose of
0.6 g was found to increase the oxygen consumption by 9%
(Ku znetsov et al., 1966). Also of interest are the authors’
observations about the body temperature rise in hu mans (in
72.3% o f the cases) by 0.2 – 0.4 C for 1 - 2 hours 1 -1.5
hours after the preparation administration.
Cellu lar hypoxia under the influence of aminothiols in
combination with the gro wth of the level o f thio l groups in
the cell due to radioprotector molecu les and endogenous
sources is an important lin k in the mechanism for
implementation of radiation protective effect of sulfur-
containing pharmaceutical drugs.
6.3. The effect of aminothiols on the cardiovascular sys-
tem
The studies by Robbers (1937) were the first to reveal that
cystamine and cisteamine possess a hypotensive action
which is accompanied by an increased systolic and minute
blood volume and a decreased peripheral vascular resistance
(Lecomte, 1952; Bella & Bacq, 1953; Mundy et al., 1960,
1963; Beau mariage et al., 1966; Bacq et al., 1968). The
mechanis m for the hypotensive action of cystamine is related
to its ability to b lock the tissue transglutaminase activity
(Engholm et al., 2011). This property of cystamine is basis
for its application for treatment of neurodegenerative
disorders (Tremblay et al., 2006). Recently, new family of
aminothiols has been discovered (Copp et al., 2011,
2013;Peebles et al., 2012) which don’t decrease AP (Soref et
al., 2012).
In hu mans, oral ad min istration of cysteamine at a dose of
0.4 – 1.2 g was revealed to cause a slight decrease in the
maximu m and minimu m arterial b lood pressure by 5 - 10
mm Hg 30 - 60 minutes after the drug administration
(Ku znetsov et al., 1961). This effect is more pronounced in
the individuals who have high blood pressure.
Admin istration of amifostine to the patients as intravenous
infusions at doses of 500 - 600 mg/ m2 of the body surface
area (concentration of the drug is more than 60 mg / liter of
blood) results in ~ 10-20% of the cases in the severe
hypotensive reaction for 5 min acco mpanied by a drop in
systolic and d iastolic pressure by 20 - 45 mm Hg and 10 - 30
mm Hg, correspondingly, up to the level of 60/ 40 mm Hg
(Turrisi et al., 1983).
6.4. Effect on the function of the gastrointestinal tract
Aminothiols at high doses when internally ad ministered in
humans cause dispeptic phenomena, sometimes the emetic
response and diarrhea. Such reactions are observed following
oral and intravenous drug admin istration. Cystamine g iven
orally at a dose of 0.8 g caused the side effects man ifesting
themselves in the form of dyspeptic symptoms (nausea,
diarrhea) in 292 subjects and headache in 84 subjects (28.8%)
(Ku znetsov et al., 1961). A mifostine when administered
orally at the doses of 800 mg and intravenously at the doses
of 200 mg/ m2 of the body surface area caused nausea and
emet ic response. ED50 of ethyol ad min istered to the patients
by drip infusions is close to 500-750 mg/ m2 of the body
surface area by its emetic act ion. The emet ic response is
observed in 25-30% of the patients when ethyol is
administered at a therapeutic dose of 300-350 mg/ m2 of the
body surface area. Repeated emet ic response within 3 hours
after the drug administration is possible (Turrisi et al.,1983).
New family of aminothiols has been discovered (Copp et al.,
2011, 2013; Peebles et al., 2012) wh ich don’t caused
vomiting (Soref et al., 2012).
It is important fro m a practical point of view to know that
cystamine inhibits the passage of the contrast material
through the intestines, which is most pronounced when it is
administered orally (Saksonov & Chernenko, 1959).
 Journal of Radioprotection Research(2014) 15-36
Aminothiols have been found to slow down the evacuation
of the contrast material fro m the stomach due to the
prolonged spasm of the pylorus and prepyloric part of the
stomach (Krasnykh, 1971).
Cystamine stimulates the secretion of salivary, gastric
and pancreatic glands (Bocharova & Mozzhukhin, 1972).
Cystamine and amifostine increase the accumulation of fluid
in the stomach when a concentration of enzymes, acid and
bicarbonates is decreased.
6.5. Effect on renal function
The decrease in the urine output was not revealed when
cystamine was administered orally in humans (Fischer,
1953). Cystamine also had no negative effect on the kidney
function in the patients with renal impairment, no changes
were revealed in clinical variables of urine, including stress
testing (Kuznetsov et al., 1966).
6.6. Effect of aminothiols on the endocrine system
A daily intravenous admin istration of cysteamine at a dose of
100 - 500 mg for up to 10 days in hu mans was found to have
no effect on the endocrine system (Kuznetsov et al., 1966).
Cysteamine can reduce the level of somatostatine in the CNS
and prolactin in the pituitary gland and blood (Millard et al.,
1985).
When amifostine is ad ministered intravenously, clin ical
man ifestations of hypocalcaemia in the form of muscle
twitches in the region of wrists and feet are possible. The
mechanis m of this effect is associated with the direct action
of the pharmaceutical preparat ion on the tissue of
parathyroid gland, causing the inhibit ion of its hormone
secretion and increasing calcium excret ion in urine ( Glover
et al., 1983).
6.7. Effect on the blood system
A leukocytic reaction is observed in humans 3.5 hours after
administration of cystamine (the growth of leukocytes up to
15.22%) (Ku znetsov et al., 1966). Accu mulation of the side
effects on the blood with the increased incidence rate of
leucopenia (fro m 12.5 to 70.6%) is possible in the clin ical
practice when amifostine is used in the long-term radiat ion
chemotherapy treatment of cancer patients (Trog et al., 1999).
In the in vitro experiments, amifostine admin istered at a
concentration of 0.1 to 1000 mcM accelerates the growth of
colonies in hu man bone marrow cells of the myeloid,
erythroid, megakaryocyte and macrophage lineages, as well
as enhances the effect of IL-1 and IL-3 on their format ion
(List et al., 1998; Ramdas et al., 2003). A stimu lating effect
of aminothiols on the hematopoietic system plays an
important role in the realization of radiat ion protective and
cytoprotective properties of the drug. Also revealed is
upregulation of thymidine kinase messenger RNA and its
transcription under the action of amifostine, wh ich may play
an important role in post-radiation recovery processes of
radiosensitive tissues (Woloschak et al., 1995).
27
6.8. Biochemical reactions to the administration of amino-
thiols
Biochemical studies of blood in the hu mans after
administration of cystamine revealed no pathological shifts.
Amifostine when ad ministered intravenously causes
hypocalcaemia up to the level of 7.5 mg of calciu m per 100
ml of b lood for 2.5 hours accompanied by 2 - 3-fold increase
in urinary calciu m excretion (up to 10.7 mM/ min) (Tu rrisi et
al., 1983; Glover et al., 1983). Hypocalcaemia is associated
with the decrease in the b lood of the levels of the thyroid
hormones and magnesium (fro m 1.9 to 1.5 mg/ 100 ml).
Cystamine, being a tissue-specific transglutaminase inhibitor,
causes the experimental reduction of neurodegenerative
changes in the brain in the case of the brain haemorrhage
(Okauchi et al., 2009) or in the patients with Parkinson’s
disease (Tremblay et al., 2006). Cystamine also inhibits
caspase activity (Lesort et al., 2003). The mechanis m of this
phenomenon is related to the ability of cystamine to bind
transglutaminase mo lecules together (Jeitner et al., 2005).
Cystamine causes inhibition of the DNA synthesis and cell
division (Murley et al., 1995). W R1065 robustly activated
the Tip60 acetyltransferase, which is a key upstream
regulator of the ATM kinase.Direct regulat ion of Tip 60's
acetyltransferase activity by WR1065 makes a significant
contribution to the radioprotective effects of WR1065 ( Xu et
al., 2011).
6.9. Effects on the function of the central nervous system
In humans, cystamine does not cause any side effects or
affect their functional activit ies in everyday life (Kuznetsov
et al., 1961). In the course of the operator’s activity for
tracking the target, the velocity of the motor reaction after
administration of cystamine at a dose of 1.2 g is not
disturbed; however, introduction to the circuit of the inertial
delays in the realization of management leads to disturbances
in the forecasting processes of the situation (Vasin &
Lebedeva, 1975).
Amifostine when ad min istered intravenously in humans
at the doses of 500 mg/ m2 of the body surface area causes
sleepiness in 20 - 30% of the cases (Torrisi et al., 1983).
7. Pharmacodynamics of Aminothiols
as Radioprotectors
The manifestation of radiation protective effect of
aminothiols persists for a defin ite period of time without any
essential changes because of the maintenance of a higher
concentration of radioprotectors in radiosensitive tissues than
in the blood; in other words, there is a temporal p lateau in
the action of the radioprotector. Aminothiols can implement
their radiation protective act ion when their intracellu lar
concentration is in the range between 0.5 - 1 mM (Swenberg
et al., 1997).
An important pharmacodynamic characteristic of
aminothiols is a d irect relat ionship between the effect iveness
28 Journal of Radioprotection Research(2014) 15-36

of a radioprotector and the dose of the preparation (Doherty, some hours (Vasin & Antipov 1972) (Figure 4).
1960; Koch, 1967; Vasin et al., 1970; Hasegawa et al., 1970)
1.6
(Figure. 3).
1.5
1.5
y = 0,0033x + 1
1.4
1.4 1
2
1.3
1.3 3

1.2 1.2
4

1.1 1.1

1 1
0 1 2 3 4 5 7 10 24
0 50 100 150

Fig. 3. Dose-response relationship of radiation protective Fig. 4. Potentiation of radiation protective effect of cysta-
effect of cystamine (Vasin et al., 1970b) mine administered to mice twice before exposure to radiation
(Vasin & Antipov 1972).
Abscissa: the dose of cystamine, mg/kg. Ordinate: radiation protec-
tive effectiveness of cystamine, DRF. Abscissa: the time interval between administration of two doses of
cystamine, hours; value 0 stands for a single administration of the
Radioprotectors from the family of aminothiols reduce drug. Ordinate: radiation protective effectiveness of cystamine,
radiation damage of the hematopoietic tissue (Devik et al., DRF: 1 - 150 mg / kg, 2 - 100 mg / kg, 3 - 75 mg / kg, 4 - 50 mg /
1955; Dup lan et al., 1966; Smith et al., 1966; Ju raskova et al., kg. Note: cystamine was administered intraperitoneally, with the
1967; Yuhas, 1969, 1970; Vasin et al., 1977), intestines second dose injected 5-10 minutes prior to γ-irradiation.
(Yuhas, 1971), skin (Lo wy & Baker, 1972;Baker et al., 1975;
Table 1. Radiation protective properties of cystamine (200
Denekamp et al., 1982; Verly et al., 1983), salivary g lands
mg/kg) during its repeated oral administration to mice ex-
(Utley et al., 1978;Sodicoff et al., 1978;Menard et al.,
posed to fractionated γ-irradiation (Vasin et al., 1991)
1984;Trog et al., 1999), heart (Kruse et al., 2003), testis
(Po merantseva & Viklina, 1975) and other tissues, have Group n Survival, % LS, day
practically no influence on the manifestation of cerebral 10 Gy 100 11,0 12,9
Cystamine + 10 Gy 100 48,0* 14,4
syndrome of ARS (Yuhas, 1969). 6 Gy 3 times every 7 days
30 3,3 12,1
The highest activity of radioprotectors takes place in the Cystamine + 6 Gy 3 times
40 70,0* 18,3
hematopoietic t issue and salivary g lands (DRF = 1.5 - 2.0), every 7 days
2.9 Gy 7 times every other day
to a lesser extent in the skin (DRF up to 1.5), intestines (DRF Cystamine + 2.9 Gy 7 times
60 3,3 8,3
up to 1.3) and testis (DRF = 1.1 - 1.2). 60 28,3* 9,4
every other day
The implementation of the radiation protective effects of 2.0 Gy 10 times every day
140 5,7 7,6
Cystamine + 2 Gy 10 times
radioprotectors is primarily associated with the reduced every day
40 5.0 4,7*
radiation damage to the stem cells of the bone marrow
Note: * - P < 0.05 versus control to irradiation by Fisher exact test,
(Duplan et al., 1966;Smith et al., 1966; Vasin et al., 1977). n – number of mice, LS – average life span of deceased animals up
Co mparative studies in vitro showed that DRF of amifostine to 30 d after irradiation
for the reduction of post-radiation death of the cells forming Table 1 shows the data on the reduction of radiation
colonies of myeloid, megakaryocyte and erythroid lineages, protective properties of cystamine ad ministered repeatedly
as well as bone marrow fibroblasts was close to 2 (Ramdas et during exposures to fractionated radiation with the reduced
al., 2003). interval between radiation fractions and the administration of
Aminothiols also have low radiat ion protective effects the radioprotector fro m 7 to one day. As can be seen, the
when they are administered following exposures to radiation
effectiveness of cystamine ad min istered weekly was rather
(Shashkov et al., 1971). The mitigatory effect of aminothiols
expressed. The reduction of the interval between radiation
robustly increases in the condition of non-uniform irradiat ion,
fractions and administration of the drug up to 2 days resulted
especially with the partial shielding of abdomen (Maisin et in the ~ 2-fold decrease of rad iation protective properties of
al., 1953a,b). An important property of aminothiols, wh ich the radioprotector.
can affect the processes of post-radiation apoptosis and Cystamine ad ministered daily was ineffective and caused
thereby contribute to a post-radiation repair of rad iosensitive a more sever development of acute rad iation sickness with
tissues, is the ability to activate the expression of NF-kappaB the reduction of an average life span of the experimental
and MnSOD genes (Murley et al., 2001, 2006). These animals by 1.5 times (Table 1). A similar effect of a more
processes are probably a foundation of the prolonging and severe course of acute radiation sickness in mice was
the potentiating of radioprotective action of cystamine over
observed by Bacq et al. (1966) when giv ing the animals
some hours at repeated its ad min istration with an intervals in
exposed to a low-intensity γ-irradiat ion within 10 days a 0.5%
 Journal of Radioprotection Research(2014) 15-36 29

mixture of cystamine and forage . 0.8 g can cause dyspeptic phenomena (nausea, vomiting,
Daily parenteral ad min istration of amifostine diarrhea, and abdominal spasms), hypotensive response
(radioprotector WR -2721) in mice exposed to fractionated accompanied by the values of blood pressure of as lo w as
radiation during 4-5 days is revealed to reduce its radiation 60/ 40 mm Hg, rare ly fever, skin reactions, anaphylaxis,
protective properties: DRF of the radioprotector was twitches in the region of the wrists and feet, transient renal
decreased from 2.3 during a single exposure up to 1.3 – 1.45 dysfunctions (Czerwinski et al., 1972).
during a repeated exposure to radiat ion (Trav is et al., 1988; The therapeutic range for use of aminothiols defined by
Coia et al., 1989). It was also observed that amifostine the relationship between the average lethal toxic dose and the
administered to the animals exposed to fractionated radiat ion average effective dose of the radiation protective effect
had a decreased effectiveness according to the test of post- (LD50/ED50) is rather limited and has a tendency to its
radiation delay of the bone growth in young animals decrease fro m s mall laboratory animals to larger ones (Table
(Damron et al., 2001). 2).
The reduced radiation protective effect of cystamine Table 2. Therapeutic index [LD50 /ED50 ] of aminothiols ad-
administered to the individuals exposed to fractionated ministered parenterally (Vasin et al., 1971; Terekhov et al.,
radiation is most likely to be associated with the 1976; Piper et al., 1979; Vladimirov et al., 1993).
accumulat ion of somatogenic phase of toxicity of the drug Mice Rats Dogs
against the background of radiation. The increased cytostatic Drugs
LD50 TI LD50 TI LD50 TI
effect of the drug and post-radiation mitotic inhib ition of Cystamine 400 4 135 1,5 73.3 1,5
bone marrow cells can serve an example of such processes, Mercaptopropylamine 850 4 400 2 125 1,4
which can dramatically reduce or co mpletely suppress the AET 470 5 322 3 100 1,6
Cystafos 950 6 260 4 - -
post-radiation repopulation of radiosensitive tissues and, thus, Aminofostine 525 9 360 5 321 4
eliminate the possibility of manifestation of radiat ion
Note:LD 50 – average lethal dose of drugs, ED 50– average effective
protective effect of the radioprotector.
(radioprotective) dose of drugs
It should be taken into account that there is a possibility
8. Toxicity, Tolerability and Side Ef- of the increased cumulat ive to xic co mp licat ions caused by
fectsof Aminothiols aminothiols administered during the exposure to ionizing
radiation, as well as in a course of chemotherapy treatment,
In humans, intravenous administration of AET at a dose of which can lead not only to a decrease in the tolerance to the
10 mg / kg was observed to cause nausea, vomiting, and pharmaceutical drugs, but also suppress the manifestation of
tachycardia (Andrews et al., 1959). Intravenous their radiation protective properties and, eventually, the
administration of cysteamine at a dose of up to 500 mg effectiveness of the whole therapeutic scheme of treatment.
during 2 to 10 days was satisfactory tolerated by patients
(Bacq et al., 1952). According to Broglie and Heinsen (1937),
8
parenteral ad min istration of cystamine (subcutaneous and
7
intramuscular) was satisfactorily tolerated at the doses of up
6
to 10 mg / kg. The drug ad min istration at the doses of 20 - 30
5
mg / kg caused an unpleasant taste in the mouth, headache,
4
and dyspeptic phenomena. Pain and the inflammatory
3
response were observed in the place of cystamine
2
administration.
1
292 subjects who received cystamine orally at a dose of 0 1 2 3 4 5 6 7 8 9 10 11 12

0.8 g observed the side effects in the form of dyspeptic

symptoms (nausea, diarrhea), and 84 subjects (28.8%) had a
headache (Kuznetsov et al., 1961).
Intravenous administration of aminofostine (ethyol) at the
doses of 300 mg/ m2 of the hu man body surface area can
cause an immediate transient hypotensive response
man ifesting itself in fainting, frequently vomit ing, more
rarely sleepiness and dizziness, rarely sneezing, hiccups,
allergic reactions with redness of the skin, eosinophilia and
an increased temperature of the body for 5 hours,
hypocalcaemia with a convulsive reaction in the region of
wrists and feet. The level of magnesium in the blood may
drop (Turrisi et al., 1983; Glover et al., 1983).
Oral ad ministration of ethyol at the doses of higher than
Fig. 5. The effect of melatonin, ascorbic and succinic acids
on the accumulation of the toxic effect of amifostine repeat-
edly administered to mice at a daily dose of 500 mg / kg for
4 days (Vasin et al., 2004).
Abscissa: the time from the beginning of the repeated administra-
tion of gammafos, days. Ordinate: mortality from intoxication, pro-
bits. ■ - succinic acid (100 mg / kg) + gammafos, + - gammafos, ◊ -
melatonin (200 mg / kg) + gammafos, ○ - ascorbic acid (200 mg /
kg) + gammafos. The arrows indicate the timing of the gammafos
administration.
Figure 5 shows the data on the presence of cumulat ive
toxicity o f amifostine repeatedly administered to mice daily
at the optimu m rad iation protective dose of 500 mg/kg
30 Journal of Radioprotection Research(2014) 15-36
(Vasin et al., 2004).
Daily ad min istration of amifostine at a dose of 300
mg/ m2 for 6 weeks to the patients with inoperable cancer in
the region of the head in the course of radiat ion
chemotherapy was observed to result in the increased
man ifestation of leukopenia up to 70.6% of the cases as
compared to 12.5% in the group without amifostine
administration (Trog et al., 1999). It was established
experimentally that the acute toxicity of cystamine during
exposures to radiation is increased according to the shift of
LD50 by 20 - 30% (Vasin et al., 1971).
One of the approaches to the prevention of cu mulat ive
toxic effects in the scheme of rad iation chemotherapy
treatment of cancer patients may be the use of natural
antioxidants (ascorbic acid, bioflavonoids, selenium, etc.),
which are known, beginning fro m the studies by Saksonov
(1968), to have the ability to reduce the to xic effects of
aminothiols (Belay et al., 1960; Ganasoundari et al., 1998).
The use of pharmaco logical agents which reduce the toxic
man ifestations of aminothiols (such as ascorbic acid, unithiol,
reduced glutathione and others) expands the boundaries of
man ifestation of a higher act ivity of a pharmaceutical
preparation with the increase in its dose, completely
sustaining a direct correlation between the radiation
protective effect and the dose of the radioprotector
(Saksonov et al., 1968;Vasin et al., 1970a).
Figure 5 shows that the use of ascorbic acid or melatonin
30 minutes before the amifostine administration reduces the
accumulat ion of the toxic effect of the radioprotector. Given
that the toxic effect of aminothiols is accompanied by the
generation of rad ical processes in the cells: 2RSH + O 2 =
RSSR + H2 O2 (Kanabus-Kaminska et al., 1988;Meier et al.,
1995).The co mbined ad ministration antio xidants and
aminothiols is pathogenetically justified (Teixeira et al.,
2003).
At present, the scientists are searching for new
radioprotectors from the group of aminothiols that do not
have the side effects such as hypotension and dyspeptic
disorders and at the same time retain high radiat ion
protective properties (Soref et al., 2012).
9. Practical Application of Sulfur-
Containing Radioprotectors
9.1. Cystamine
Cystamine dihydrochloride is a wh ite crystalline powder
with a cream shade, highly soluble in water. The drug is
taken at one time in the dose of 1.2 g 30-60 minutes before
exposure to ionizing radiation. The repeated administration
of cystamine is possible in the interval of 4 hours, which
increases the effect of the drug up to 8 hours.
Radiat ion protective properties of cystamine
administered per orally to humans were estimated by th e
reduction of chro mosomal aberrations in the bone marrow
cells in the course of radiotherapy treat ment of the cancer
patients exposed to fractionated irradiation of sternum at a
dose of 2.5 Gy. According to the results of the research
conducted on 57 patients, DRF of cystamine administered
orally to humans at a dose of 1.2 g was close to 1.2
(Vladimirov, 1982).
9.2. Amifostine
Amifostine (ethyol “Schering.Plough”, WR-2721, USA ) by
amma-aminopropylaminoethyl-
thiophosphoric acid: H2 N.(CH2 )3 HN(CH2 )2 SPO3 H2 .
It is used in chemotherapy and radiotherapy treatment of
cancer patients to reduce the radiation damage to healthy
tissues which are exposed to ionizing radiation in the
radiotherapy treatment of tu mors, as well as to protect them
fro m the cytotoxic effects of chemo-therapeutic agents
(alkylating agents, mitomycin C, platinum preparations).
In the practice of chemotherapy for treat ment of cancer
patients, amifostine makes it possible to reduce the cytotoxic
effect of chemotherapeutic drugs on the bone marrow(Glick
et al., 1984; Glover J. et al., 1986, 1987; Poplin et al., 1994;
Capizzi et al., 1995; Betticher et al., 1995; Kemp et al.,
1996).
The first clinical studies of amifostine in radiotherapy
treatment of cancer patients provide evidence that the
radioprotector reduces radiation damage of skin, intestine,
lung, pelvis and salivary glands (Takahashi et al., 1986;
Kligerman et al., 1992; McDonald et al., 1994; Bohuslavizki
et al., 1999; Trog et al., 1999; Dunst et al., 2000; Antonadou
et al., 2001; Koukourakis et al., 2009, 2011; Katsanos et al.,
2010), and don’t decrease anticancer effect of radiotherapy
(Bourhis et al., 2011).
Much attention has been drawn in recent years to the
ability of amifostine to significantly reduce the incidence of
complications as a result of radiotherapy for treat ment of
head and neck cancer patients, which are associated with
radiation damage to salivary glands and manifest xerostomia
(McDonald et al., 1994; Brizel et al., 1998; Bohuslavizki et
al., 1999; Trog et al., 1999).
7
1
6
5 2
4
3
2
15 25 35 45 55
Fig. 6. Radiation protective effect of ethyol (300 mg/ m2 ) in
humans on the reduction of the symptoms of post-radiation
mucositis during the radiotherapy treatment (calculated by
the author based on the data provided by Trog et al., 1999).
Abscissa: the cumulative dose during the fractionated radiotherapy,
Gy Ordinate: the frequency of manifestation of post-radiation mu-
cositis, probits. 1 - radiotherapy alone, 2 - ethyol+radiotherapy;
ED50 for control to radiotherapy alone is 34.9 Gy (y = 7.09 D – 5.94,
 Journal of Radioprotection Research(2014) 15-36
r = 0.97 p <0.01). ED 50 for group treated with ethyol+radiotherapy
is 47.7 Gy(у = 6.72D – 6.28, r = 0.93, P <0.05). DM F for amifos-
tine is 1.37.
Clin ical findings by Trog et al. (1999) make it possible to
define that DRF of ethyol by the reduction of the severity of
the damage to the salivary glands in hu mans is equal to the
value of 1.37. (Figure 6). Positive effects of subcutaneous
administration of aminofostine have been observed in
hypofractionated radiotherapy for treat ment of breast cancer
(Koukourakis I. et al., 2009).
According to the meta-analysis, admin istration of
aminofostine for treat ment of more than 1,000 patients with
lung, head neck, and cervical cancers did not reduce the
effectiveness of radiochemotherapy (Bourhis et al., 2011).
10. Conclusion
Over the years, extensive experimental studies of radiation
protective agents fro m aminothiols series were co mp leted by
discovery of radioprotectors cystamine and amifostine used
in med ical practice. Characteristic features of the mechanism
of radiation protective action of aminothiols unlike other
antioxidants are potential chance of the development of
complete neutralization of o xygen effect as radiolog ical
phenomenon, first of all, in DNA rad iolysis. This peculiar
action of aminothiols is connected with their chemical
structure being able to squeeze DNA helix v ia diamine bond
of aminothiol d isulfide thereby decreasing availabilility of
critical points of DNA molecu les to radicals of o xygen
species. Positive charge of aminothiol mo lecules has an
advantage for the development of high their concentration in
negative DNA mo lecules. In addition, pharmacolog ical
properties of aminothiols can deierminate co mp lex
polyfunctional action which is significant for the radiat ion
protection, including the influence to mithochondria
respiration following acute hypoxia, gene Mn-SOD
expression, etc. However, the decrease of DNA damage by
aminothiols is basic factor in their protective action. The
properties of aminothiols to block transaminase following a
development of hypotension expand their application in
clin ical practice and the same thing is a cause for severe side
effect. Character of aminothiol pharmadinamics is necessary
to take into consideration in its repeated application in the
scheme of in radiotherapy.
Acknowledgment
The research cited in this article was fro m publicat ions
through April 2014. The opin ion contained in this paper is
the view of the author.
Declaration of Interests
The author declare no conflicts of interest and is alone
responsible for the content and writing of this paper.
31
References
Akerfeldt, S., Ronnback, C., Nelson, A. (1967). Radioprotective agents:
results with S-(3-amino-2-hydroxypropyl)phosphorothioate, ami-
dophosphorothioate and some related compounds. Radiation Research,
31, 850-855.
Akerfeldt, S. (1959). Preparation and determination of sodiumhydrogen S-
(2-aminoethyl)-phosphorthioate. Acta Chemica Scandinavia,13,1479-
1480.
Akerfeldt, S. (1963).Radioprotective effects of S-phosphorylated thiols.Acta
Radiologica,45, 465-469.
Alexander, P., Charlesby, A. (1954). Physico-chemical methods of pro-
tection against ionizing radiations.Radiobiological Symposium Liege.
London: Butterworth (pp. 49-59).
Allalunis-T urner, M. J., Walden, T. J., Sawich, C. (1989).Induction of mar-
row hypoxia by radioprotective agents.Radiation Research,118,581-
586.
Allalunis-T urner, M. J. (1990). Reduced bone marrow pO 2 following treat-
ment with radioprotective drugs. Radiation Research,122,262-267.
Alper, T ., Howard-Flanders, P. (1956).The role of oxygen in modifying the
radiosensitivity of E.coli B. Nature178, 978-979.
Andrews, J. R., Sneider, S. E. (1959). The modification of the radiation re-
sponse.American Journal of Roentgenology,81(3), 485-493.
Antonadou, D., Coliarakis, N., Synodinou, M., Athanassiou, H., Kouveli, A.,
Verigos, C., Georgakopoulos, G., Panoussaki, K., Karageorgis, P.,
T hrouvalas, N.,Clinical Radiation Oncology Hellenic Group. (2001).
Randomized phase III trial of radiation treatment +/- amifostine in pa-
tients with advanced-stage lung cancer. International Journal of Ra-
diation Oncology Biology Physics,51, 915-922.
Arbuzov, S. Ia., Vazanov, V. A., Nekachalova, I. Ia., Patalova, V. N., Pete-
lina, V. V., Shamova, E. K. (1961). The distribution of sulphur of
mercamine in organs and tissues of irradiated and non-irradiated ani-
mals.Memoires de la Academia des Chirurgiens de Paris, 6, 62-66.
Ariens, E. J., van Rossum, J. M., Simonis, A. M. (1957). Affinity, intrinsic
activity and drug interactions.Pharmacological Review, 9(2), 218-236.
Ashwood-Smith, M. J. (1961).The radioprotective action of dimethyl-
sulphoxide and various other sulphoxides.International Journal of
Radiation Biology,3(1), 41-48.
Bacq, Z. M., Alexander, P. (1955).Fundamentals of radiobiology. London:
Butterworth.
Bacq, Z. M.,Beaumariage, M. L., Goutier, R., Van Caneghem, P. (1968).
T he state of shock induced by cystamine and cysteamine. British
Journal of Pharmacology,34, 202-203.
Bacq, Z. M., Caneghem, P. (1966). The influence of cystamine administered
by mouth to mice irradiated with gamma-rays at a low dose-rate. In-
ternational Journal of Radiation Biology, 10,595-599.
Bacq, Z. M., Herve, A. (1952). The protective effect of amines against x-
irradiation.J Physiol, 118(2), 24-25.
Bacq, Z. M., Herve, A., Lecomte, J., Fischer, P. (1950). Cyanide Protection
against X-Irradiation. Science,111(2884), 356-357
Bacq, Z. M., Herve, A., Lecomte, J., Fischer, P., Blavier, J. (1951).
Protection contre le rayonnement X par la beta-mercaptoethylamine.
Archives Internationales de Physiologie, Biologie et de
Biophyysique,59, 442-447.
Baker, D. G., Leith, J. T. (1975). Protection of the skin of mice against irra-
diation with cyclotron accelerated Helium ions by2-beta-
mercaptoethylamine. Acta Radiologica, 14, 561-571.
Barron, E., Dickman, S., Muntz, I., Singer, T. R. (1949).Studies on the
mechanism of action of ionizing radiatuins.Journal of General Physi-
ology, 32(4), 537-552.
Beaumariage, M. L., van Caneghan, P., Bacq, Z. M. (1966). Study of phar-
macodinamic properties of cystamine and cysteamine in different an-
imal species.Strahlentherapie,131, 34-51.
Bella, D. D., Bacq, Z. M. (1953). Effect of cysteamine on isolated frog heart
and effect of vagal excitation in turtle.Naunyn Schmiedebergs Arch
Exp Pathol Pharmakol, 219(4), 366-370.
Belaya, V. E., Vasil’ev, P. V., Saksonov, P. P. (1960).Data to comparative
pharmacological characteristics of various salts of mer-
camine.Farmakologiia i Toksikologiia, 23, 450-453.
Benson, R. E., Michaelson, S. M., Downs, W. (1961). T oxicological and
radioprotection studies on S,beta-aminoethylisothiouronium bromide
(AET ). Radiation Research, 15, 561-567.
32 Journal of Radioprotection Research(2014) 15-36
Betticher, D. C., Anderson, H., Ranson, M., Meely, K., Oster, W., Thatcher,
N. (1995). Carboplatin combined with amifostine, a bone marrow pro-
tectant, in the treatment of non-small-cell lung cancer: a randomised
phase II study.British Journal of Cancer, 72(6), 1551-1555.
Bocharova, M. A., Mozzhukhin, A. S. (1972). The influence of cystamine to
secretion of gastro-intestinal tract.Farmakologiia i Toksikologi-
ia,35,310-315
Bohuslavizki, K. H. (1999). Salivary gland protection by amifostine in high-
dose radioiodine therapy of differentiale thyroid can-
cer.Strahlentherapie und Oncologie, 175, 57-61.
Bott, K. F., Lundgren, D. G. (1964).The relationship of sulfhydryl and disul-
fide constituents of bacillus cereus to radioresistance.Radiation Re-
search,21,195-211.
Brizel, D. M.,Movsas, B., Buentzel, J., Langendijk, J. A., Komaki, R., Swan
Leong, S., Levendag, P., Pignon, J. P. (2011). Effect of amifostine on
survival among patients treated with radiotherapy: a meta-analysis of
individual patient data. Clinical Oncology, 29,2590-2597.doi:
10.1200/JCO.2010.33.1454.
Brizel, D. M. (1998).Radiotherapy and concurrent chemotherapy for the
treatment of locally advanced head and neck squamous cell carcino-
ma.Seminars in Radiation Oncology,8(4), 237-246.
Broglie, M., Heinsen, H. A. (1937).Zur Frage der Cystaminwirkung auf
Blutdruck und Blutzucker bei Menschen.Archiv fur Experimentelle
Pathologie und Pharmakologie, 188,279-288.
Brown, P.E. (1967). Mechanism of action of aminothiol radioprotec-
tors.Nature,13,363-364.
Calabro-Jones, P. M., Fahey, R. C., Smoluk, G. D., Ward, J. F. (1985).
Alkaline phosphatase promotes radioprotection and accumulation of
WR-1065 in V79-171 cells incubated in medium containing WR-2721.
International Journal of Radiation Biology,47, 23-27.
Capizzi, R. L., Oster, W. (1995). Protection of normal tissues from cytotoxic
effects of chemotherapy and radiation by amifostine. Clinical experi-
ences. European Journal of Cancer,31A (Suppl) 1, 8-13.
Coia, L. R., Brown, D. Q. (1989). Protection on bone marrow by WR-2721
after fractionated irradiation. International Journal of Radiation On-
cology Biology Physics,17(4), 908-909.
Cole, L. J., Bond, V. P., Fischer, M. (1952). Protection of mice against X-
irradiation mortality by sodium nitrite.Science,115(2998), 644-646.
Copp, R. R., Peebles, D. D., Soref, C. M., Fahl, W. E. (2013). Radioprotec-
tive efficacy and toxicity of a new family of aminothiol ana-
logs.Internatiional Journal of Radiation Biology, 89, 485-492. doi:
10.3109/09553002.2013.770579.
Copp, R. R., Peebles, D. D., Fahl, W. E. (2011). Synthesis and growth regu-
latory activity of a prototype member of a new family of aminothiol
radioprotectors.Bioorganic Medical Chemistry Letters,21,7426-
7430.doi: 10.1016/j.bmcl.2011.10.011.
Crough, B. G., Overman R. R. (1957).Chemical protection against x-
radiation death in primates: a preliminary report.Science,125(3257),
1092.
Сzerwinski, A. W., Clark, M. L. (1972). Report MCA 1-33. U.S. Army Med.
Res. Develop. Command.
Dale, W. M., Davies, J. V., Meredith, W. J. (1949). Further observations on
the protection effect in radiation chemistry.British Journal of Can-
cer,3(1), 31-41.
Damron, T. A., Margulies, B., Biskup, D., Spadaro, J. A. (2001). Amifostine
before fractionated irradiation protects bone growth in rats better than
fractionation alone.International Journal of Radiation Oncology Biol-
ogy Physics,50(2), 479-483.
Davidson, D. E., Grenan, M. M., Sweeney, T. R. (1980). Biological charac-
teristics of some improved radioprotectors. In:L.W. Brady
(Ed),Radiosensitizers. NY:Masson Publ. pp. 309
Denecamp, J., Michael, B. D., Rojas, A., Stewart, F. A. (1982). Ra-
dioprotection of mouse skin by WR-2721: the critical influence of ox-
ygen tension.International Journal of Radiation Oncology Biology
Physics,8(3-4), 531-534.
Devik, F., Lothe, F. (1955).The effect of cysteamine, cystamine and hy poxia
on mortality and bone-marrow chromosome aberrations in mice after
total bogy roentgen irradiation.Acta Radiologica,44,243-248.
Doherty, D. G., Burnett. W. T. (1955). Pritective effect of S,beta-amino-
ethylisothiouronium Br*HBr and related compounds against X-
irradiation death in mice.Proceedings of Society for Experimental Bi-
ology and Medicine, 89(2), 312-314.
Doherty, D. G. (1960).Chemical protection to mammals against ionizing
radiation. In: Hollaender A (Ed),Radiation Protection and Recovery.
New York: Pergamon Press (pp. 45-86).
Dowdy, A. H., Bennett, L. R., Chastain, S. M. (1950).Protective action of
anoxia against total body roentgen irradiation of mam-
mals.Radiology,55(6), 879-885.
Duplan, J. F., Fuhrer, J. (1966). Estimation of the radioprotective effect of
aminoethylisothiouronium (AET) by enumeration of the splenic nod-
ules.Comptes rendus des séances de la Sociétéde biologie et de ses
filiales,160(6), 1142-1145.
Dunst, J. (2000). Postoperative simultaneous radiochemotherapy improves
survival in patients with cervical carcinoma.Strahlentherapie undOn-
kologie,176(10), 486-487.
Fahey, R. C. (1988). Protection of DNA by thiols.Pharmacology and Ther-
apeutics, 39(1-3), 101-108.
Firket, H., Lelievre, P. (1966). Effect de la cystamine sur la respiration, la
phosphorylation oxydative et l`ultrastructure des mitochon-
dries.International Journal ofRadiation Biology,10(4), 403-415.
Fischer, P., Lecomte, J. (1953). Effects de la beta-mercaptoethylamine sur la
diurese provoquee par les mercuriels. Archive of internationalPhar-
macodynamics and Therapeutics, 93, 118.
Frey, R., Hagen, U. (1974). Oxygen effect on gamma-irradiated
DNA.Radiation and Environmental Biophysics.11(2), 125-33.
Edgren, M., Larsson, A., Nilsson, K., Révész, L., Scott, O. C. (1980).Lack
of oxygen effect in glutathione-deficient human cells in cul-
ture.International Journal of Radiation Biology,37(3), 299-306.
Eidus, L. Kh., Ganassi, E. R. (1959). Studies on the mechanism of after-
effect of radiations in proteins.Biofizika, 4(2), 215-223.
Eĭdus, L. Kh., Korystov, Iu. N. (1980). "Oxygen effect" mechanism in the
action of ionizing radiation on cells.Radiobiologiia,20(4), 537-541.
Eldjarn, L., Pihl, A. (1956). On the mode of action of x-ray protective agents.
I. T he fixation in vivo of cystamine and cysteamine to pro-
teins.Journal of Biological Chemistry, 223(1), 341-352.
Ganasoundari, A., Devi, P. U., Rao, B. S. (1998). Enhancement of bone
marrow radioprotection and reduction of WR-2721 toxicity by Oci-
mum sanctum.Mutation Research,397(2), 303-312.
Giles, N. H., Riley H. P. (1950).Studies on the mechanism of the oxygen
effect on the radiosensitivity of T radescantia chromo-
somes.Proceedings of National Academy of Science of USA,,36(6),
337-344.
Glick, J. H., Glover, D., Weiler, C., Norfleet, L., Yuhas, J., Kligerman, M.
M. (1984). Phase I controlled trials of WR-2721 and cyclophospha-
mide.International Journal of Radiation Oncology Biology Phys-
ics,,10(9), 1777-1780.
Glover, D., Riley, L., Carmichael, K.,Spar, B.,Glick, J., Kligerman, M. M.,
Agus, Z. S., Slatopolsky, E., Attie, M., Goldfarb, S.(1983). Hy-
pocalcemia and inhibition of parathyroid hormone secretion after ad-
ministration of WR-2721 (a radioprotective and chemoprotective
agent).New English Journal of Medicine,309(19), 1137-1141.
Glover, D., Glick, J.H., Weiler, C., Fox, K., Turrisi, A., Kligerman, M.M.
(1986). Phase I/II trials of WR-2721 and cis-platinum.International
Journal of Radiation Oncology Biology Physics,12(8), 1509-1512.
Glover, D.,Glick, J. H., Weiler, C., Fox, K., Guerry, D. (1987). WR-2721
and high-dose cisplatin: an active combination in the treatment of
metastatic melanoma.Journal of Clinical Oncology,5(4), 574-578.
Graevskii, E.Ia. (1967). On the role of sulfhydryl groups in determining
natural radiosensitivity and its artificial variation.Radiobiologiia, 7(5),
715-727.
Gray, J. L., Moulden, E. J., T ew, J. T ., Jensen, H. (1952a). Protective effect
of pitressin and of epinephrine against total body X-
irradiation.Proceedings ofthe Society for Experimental Biology and
Medicine,79(3), 384-387.
Gray, J. L., T ew, J. T ., Jensen, H. (1952b). Protective effect of serotonin and
paraaminopropiophenon against lethal doses of X-
irradiation.Proceedings ofthe Society for Experimental Biology and
Medicine,80(4), 604-607.
Gray, L. H., Conger, A. D., Ebert, M. et al. (1953).The concentration of
oxygen dissolved in tissues at t he time of irradiation as a factor in ra-
diotherapy.British Journal ofRadiology,26, 638-648.
Grdina, D. J., Shigematsu, N., Dale, P., Newton, G. L., Aguilera, J. A.,
Fahey, R. C. (1995).Thiol and disulfide metabolites of the radiation
protector and potential chemopreventive agent WR-2721 are linked to
both its anti-cytotoxic and anti-mutagenic mechanisms of ac-
tion.Carcinogenesis, 16(4), 767-774.
 Journal of Radioprotection Research(2014) 15-36
Hanson, B., Sorbo, B. (1961).Radioprotective effects of aminoalkyl thio-
esters.Acta Radiologica,56, 141-144.
Harris, J. W., Phillips, T. L. (1971). Radiobiolgical and biochemical studies
of thiophophate radioprotective compounds related to cysteamine. Ra-
diation Research, 46, 362-379.
Hasegawa, A. T ., Landahl, H. D. (1967). Studies on spleen oxygen tension
and radioprotection in mice with hypoxia, serotonin and p-
aminopropiophenone.Radiation Research ,31,389–399.
Hasegawa, A. T ., Landahl, H. D. (1970). Dose-redaction factor for radiation
lethality in mice as function of dose of mercaptoethylamine.Radiation
Research,44, 738-747.
Held, K. D. (1988). Models for thiol protection of DNA in cells.Pharmacol
Ther. 39(1-3), 123-131.
Holmberg, B., Sorbo, B. (1959). Protective effect of 2-aminoethylthiosul-
phuric acid. Nature 183, 832.
Howard-Flanders, P. (1960). Effect of oxygen on the radiosensitivity of
bacteriophage in the presence of sulphydryl compounds.Nature,186,
485-487.
Hutchinson, F. (1961). Molecular basis for action of ionizing radia-
tions.Science,134(3478), 533-538.
Iarmonenko, S. P., Rampan, Iu. I., Karochkin, B. B., Berezhnova, L. I.,
Ovakimov, V. G. (1970). Oxygen tension kinetics in critical organs
under the effects of mexamine in comparison with its radiation-
protective effect.Radiobiologiia,10(5),700-705.
Iarmonenko, S.P. (1961).Influence of cystamine to tissue respira-
tion.Radiobiologiia, 1(6), 903.
Ilyin, L. A., Rudnyi, N. M., Suvorov, N. N., Chernov, G. A., Antipov, V. V.,
Vasin, M. V., Davydov, B. I., Mikhailov, P. P. (1994). Indralin – ra-
dioprotector of urgent action: radiation–protective property, pharma-
cology, mechanism of action, clinics. Moscow: Ministry of Health
Care of Russia.
Jacobus, D. P. (1959).Preprotection of the dog against ionizing radia-
tion.Federation Proceeding 18, 74.
Jeitner, T. M.,Delikatny, E. J., Ahlqvist, J., Capper, H., Cooper, A. J. (2005).
Mechanism for the inhibition of transglutaminase 2 by cyst a-
mine.Biochemical Pharmacology, 69, 961-970.
Jellum, E. (1965).Interaction of cysteamine and cystamine derivatives with
nucleic acids and nucleoproteins.International Journal of Radiation
Biology, 9, 185-200.
Jokay, I., Kelemenics, K., Gyuris, A., Minarovits, J. (1998). S-
methylthiocysteine and cystamine are potent stimulators of thiol pro-
duction and glutathione synthesis. Life Science,62, 27-33.
Jones, D. P., Mason, H. S. (1978). Gradients of O2 concentration in hepato-
cytes.Journal ofBiological Chemistry,235(13), 4874-4880.
Juraskova, V., T kadlecek, L. (1967).The effect of the interval between in-
traperitoneal administration of cystamine and exposure on the dose-
reduction factor for hematopoietic stem cells.Radiation Reearch, 30,
14-21.
Kahn, J. J. (1951). Modification of sensitivity to X-irradiation by morphine
sulfate.Proceedings of Society for Experimental Biology andMedi-
cine,78(3), 486-488.
Kaluszyner, A., Czerniak, P., Bergman, E. D. (1961). Thazolidines and
aminoalkylthiosulfuric acid as protecting agents against ionizing radi-
ation.Radiation Research,14,23-28.
Kanabus-Kaminska, J. M., Feely, M., Birnboin, H. (1988).Simultaneous
protective and damaging effects of cysteamine on intracellular DNA
of leukocites.Free Radical Biology and Medicine, 4, 141-145.
Kandasamy, S. B., Kumar, K. S., Hunt, W. A.,Weiss, J. F.(1988). Opposite
effects of WR-2721 and WR-1065 on radiation-induced hypothermia:
possible correlation with oxygen uptake. Radiation Research,114,240-
247.
Katsanos, K. H.,Briasoulis, E., Tsekeris, P., Batistatou, A., Bai, M., T olis,
C., Capizzello, A., Panelos, I., Karavasilis, V., Christodoulou, D.,
T sianos, E.V.(2010). Randomized phase II exploratory study of
prophylactic amifostine in cancer patients who receive radical radio-
therapy to the pelvis.Journal of Experimental and Clinical Cancer
Research,29, 68.
Kemp, G. I., Rose, P.,Lurain, J., Berman, M., Manetta, A., Roullet, B.,
Homesley, H., Belpomme, D., Glick, J. (1996). Amifostine pretreat-
ment for protection against cyclophosphamide-induced and cisplatin-
induced toxicities: results of a randomized control trial in patients
with advanced ovarian cancer.Journal ofClinical Oncology,14(7),
2101-2112.
33
Kligerman, M. M., Liu, T., Liu, Y., Scheffler, B., He, S., Zhang, Z.
(1992).Interim analysis of a randomized trial of radiation therapy of
rectal cancer with/without WR-2721.International Journal of Radia-
tion Oncology Biology Physics,22(4), 799-802.
Koch, R. (1967). Analysis Untersuchungen uber einen biologischen Strah-
lenschutz. 80 Mitteilung: Uber quantitative Beziehungen des chem-
ischen Strahlenschutzes zu seinen Wirkungsmechanismus. Strahlen-
therapie, 134, 102-106.
Korst, A.E., Eeltink, C.M., Vermorken, J.B., Van der Vijgh, W.J. (1997).
Pharmacokinetics of amifostine and its metabolites in pa-
tients.EuropeanJournal of Cancer Parta,33,1425-1429.
Koterov, A. N., Filippovich, I. V. (1995). The radiobiology of metallothi-
oneins.Radiatsionnaia Biologiia Radioecologiia, 35(2), 162-180.
Koukourakis, M. I., Kyrgias, G., Papadopoulou, A., Panteliadou, M.,
Giatromanolaki, A., Sivridis, E., Mavropoulou, S., Kalogeris, K.,
Nassos, P., Milioudis, N., T ouloupidis, S.(2011). Treatment of low-
risk prostate cancer with radical hypofractionated accelerated radio-
therapy with cytoprotection (HypoARC): an interim analysis of tox-
icity and efficacy. Anticancer Research, 31(5), 1745-1751.
Koukourakis, M. I.,Tsoutsou, P. G., Abatzoglou, I. M., Sismanidou, K.,
Giatromanolaki, A., Sivridis, E. (2009). Hypofractionated and accel-
erated radiotherapy with subcutaneous amifostine cytoprotection as
short adjuvant regimen after breast-conserving surgery: interim re-
port.International Journal of Radiation Oncology Biology Physics, 74,
1173-1180
Krasnykh, I. G., T Iutin, L. A. (1971). Influence of some radioprotective
agents to motive evacuative function of gastric intestinal tract. In:
Saksonov PP, Davydov BI, eds, Radiobiological aspects of the reac-
tivity of organism during space flights. Problems of space biology.Vol.
14. Moscow: Nauka. 1971. English translation accessed 1973 by
NASA T T F–721. Washington DC: National Aeronautics and Space
Administration (pp. 57–65).
Kruse, J. J., Strootman, E. G., Wondergem, J. (2003). Effects of amifostine
on radiation-induced cardiac damage.Acta Oncology, 42(1), 4-9.
Kuznetsov, V. I., Kushanovskii, M. S., Mikhasev, M. I. (1961). On antihy-
pertensive action of cystamine.Clinicheskaia Meditsina, 39, 71-76.
Kuznetsov, V. I., T ank, L. I. (1964).The change of oxygen consumption of
erythrocytes by cystamine.Radiobiologiia, 4, 284-288.
Kuznetsov, V. I., T ank, L. I. Pharmacology and clinic application of amino-
thiols. Moscow: Meditsina, 1966.
Langendorff, H., Koch, R. (1956). β-Aminoäthylisothiuronium als peroral
wirksame Strahlenschutzsubstanz. Naturwissenschaften, 43, 534-535.
Lea, D. E. (1947). T he action of radiations on dilute aqueous solutions: the
spatial distribution of H and OH. British Journal of Radiology, Suppl.
(1), 59-64.
Lecomte, J. (1952). Proprietes pharmacodynamiques de la cystinamine.
Archives Internationales de Physiologie, Biologie et de Biophyysique,
60, 179-180.
Lelievre, P. (1965). Action de la cysteamine sur la consommation d`oxygene
et phosphorylation oxydetive couplee mitochondries de foie de rat. In-
ternational Journal of Radiation Biology, 9, 107-113.
Lehninger, A., Schneider, M. (1959). Mitochondrial swelling induced by
glutathione.The Journal of Biophysical and Biochemical Cytology, 5,
109-115.
Lesort, M., Lee, M., Tucholski, J., Johnson, V. W. (2003). Cystamine inhib-
its caspase activity. Journal of Biological Chemistry, 278, 3825-3830.
Limperos, G., Mosher, W. A. (1950). Protection of mice against X-irra-
diation by thiourea.Science, 112, 86-87.
List, A. F., Heaton, R., Glinsmann-Gibson, B., Capizzi, R. L. (1998).
Amifostine stimulates formation of multipotent and erythroid bone
marrow progenitors. Leukemia, 12, 1596-1602.
Lowy, R. O., Baker, D. G. (1972). Protection against local irradiation injury
to the skin by locally and sistematically applied drugs. Radiology,105,
425-428
Maisin, J., Mandart, M., Lambert, G., Maisin, H. (1953a).Curative action of
beta-mercapto-ethylamine in the rat irradiated with the liver protected.
Comptes rendus des séances de la Sociétéde biologie et de ses filiales,
147, 362-364.
Maisin, J. H., Lambert, G., Mandart, M., Maisin, H. (1953b).Therapeutic
action of glutathione and beta-mercaptoethylamine against a lethal
dose of x-rays.Nature, 171, 971.
Мaisin, J. H., Maisin, H., Dunjic, A. (1954). Influence de l`injection de
mercaptoethylamine avant l`irradiation sur la survie des animaux
34 Journal of Radioprotection Research(2014) 15-36
injectes apres l`irradiation d`une suspension de moelle osseuse.
Comptes rendus des séances de la Sociétéde biologie et de ses filiales,
148, 1293-1297.
Maisin, J. H., Matterin, G., Fredman-Manduzio, A., van den Porren, L.
(1968). Reduction of short and longtern radiation lethality by mixtures
of chemical protectors.Radiation Research, 35, 26-44.
Mayer, S. H., Patt, H. M. (1953). Potentiation of cysteine protection against
X-radiation by dinitrophenol or hypoxia.Federation Proceed-
ings,12(2), 94-95.
McCord, J. M., Fridovich, I. (1978). The biology and pathology of oxygen
radicals.Annals of Internal Medicine, 89(1), 122-127.
McDonald, S., Meyerowitz, C., Smudzin, T ., Rubin, P. (1994). Preliminary
results of a piot study using WR-2721 before fractioned irradiation of
the head and neck produce salivary gland dysfunction. International
Journal of Radiation Oncology Biology Physics,29, 747-754.
Meier, T ., Issels, R. D. (1995).Degradation of 2-(3-aminopropylamino)-
ethanethiol (WR-1065) by Cu-dependent amine oxidases and influ-
ence on glutathione status of Chinese hamster ovary cells.Biochemical
Pharmacology,50,489-496.
Menard, T . W., Izutsu, K. T., Ensign, W. Y., Keller, P. J., Morton, T. H.,
T ruelove, E. L. (1984). Radioprotection by WR-2721 of gamma-
irradiated rat parotid gland: effect on gland weight and secretion at 8-
10 days post irradiation.International Journal of Radiation Oncology
Biology Physics,10(9),1555-1559.
Midander, J., Deschavanne, P. J., Malaise, E. P., Revesz, L. (1982). Survival
curves of irradiated glutathione-deficient human fibroblasts: induction
of a reduced enhancement of radiosensitivity by oxygen and misoni-
dazole.International Journal of Radiation Oncology BiologyPhys-
ics,8(3-4), 443-446.
Millard, W.J., Sagar, S.M., Martin, J.B. (1985). Cysteamine-induced deple-
tion of somatostatin and prolactin.Fediration Proceedings,44,2546-
2550.
Mole, R. H., Philpot, J. S., Hodges, C. R. (1950).Reduction in lethal effect
of X-irradiation by pretreatment with thiourea or sodium-
dithiophosphonate.Nature,166,515.
Morse, M. L., Dahl, R. H. (1978). Cellular glutathione is a key to the oxy-
gen effect in radiation damage.Nature, 271(5646), 660-662.
Mundy, R., Heiffer, M., Mehlman, B. (1963).Mechanism of beta-mer-
captoethylamine-induced hypotension in the dog.American Journal of
Physiology,204, 997-1000.
Mundy, R. L., Heiffer, M. H. (1960). The pharmacology of radioprotectant
chemicals.General pharmacology of -mercaptoethylamine.Radiation
Research, 13, 381-394.
Murley, J. S., Grdina, D. J. (1995). The effect of cycloheximide and WR-
1065 on repair processes: a mechanism for chemoprotection. Carcin-
ogenesis,16, 2699-2705.
Murley, J. S., Kataoka, Y., Weydert, C. J., Oberley, L. W.,Grdina, D. J.
(2006). Delayed radioprotection by transcription factor kB-mediated
induction of manganese superoxide dismutase in human microvascu-
lar endothelial cells after exposure to the free radical scavenger
WR1065.Free Radical Biology and Medicine, 40, 1004-1016.
Murley, J. S., Kataoka, Y., Baker, K. L., Diamond, A. M., Morgan, W. F.,
Grdina, D. J. (2007). Manganese superoxide dismutase (SOD2)-
mediated delayed radioprotection induced by the free thiol form of
amifostine and tumor necrosis factor alpha.Radiation Research,
167(4), 465-474.
Nasonov, D. N., Aleksandrov, V. Ia. (1940). Reaction of viable substance on
external influence. Akademiia Nauk SSSR. Moscow.
Neubert, D., Lehninger, A. (1962). The effect of thiols and disulfides on
water uptake and extrusion by rat liver mitochondria.Journal of Bio-
logical Chemistry, 237, 952-958.
Newton, G. L., Aguilera, J. A., Kim, T., Ward, J. F., Fahey, R. C. (1996a).
T ransport of aminothiol radioprotectors into mammalian cell: Passive
diffusion versus mediated uptake. Radiation Research,146, 206-215.
Newton, G. L., Aguilera, J. A., Ward, J. F., Fahey R. C. (1996b). Binding of
radioprotective thiols and disulfides in Chinese hamster V 79 cell nu-
clei. Radiation Research,146,298-305.
North, S., El-Ghissassi, F., Pluquet, O., Verhaegh, G.,Hainaut, P.
(2000).The cytoprotective aminothiol WR1065 activates p21waf-1
and down regulates cell cycle progression through a p53-dependent
pathway. Oncogene, 19, 1206-1214.
Okauchi, M., Xi, G., Keep, R. F., Hua, Y. (2009). Tissue-type transglutami-
nase and the effects of cystamine on intracerebral hemorrhage-
induced brain edema and neurological deficits. Brain Re-
search,1249,229-236.
Ormerod, M. G., Riesz, P. (1967). An unusual reaction of hydrogen sulphide
and hydrogen iodide with free radicals in irradiated dry deoxyribonu-
cleic acid.Biochimical and Biophysical Acta, 149(2), 451-458.
Paterson, E., Matthews, J. (1951). Protective action of ethyl alcohol on ir-
radiated mice.Nature,168(4287), 1126-1127.
Patt, H. M., Mayer, S., Straube, R., Jackson, E. (1953). Radiation dose re-
duction by cyseine.Journal of Cellular and Comparative Physiology,
42, 327-331.
Patt, H. M., Tyree, E. B., Straube, R. L., Smith, D. E. (1949). Cysteine
protection against X-irradiation.Science, 110, 213-214.
Peebles, D. D.,Soref, C. M., Copp, R. R., T hunberg, A. L., Fahl, W.
E.(2012). ROS-scavenger and radioprotective efficacy of the new
PrC-210 aminothiol.Radiation Research, 178, 57-68.
Piper, J., Rose, L., Johnston, T. P., Grenan, M. M. (1975). Hydroxy de-
rivatives of S-2-(3-aminopropylamino)ethyl dihydrogen phos-
phorothioate and related compounds as antiradiation agents. Journal
of Medical Chemistry,18, 803-812.
Piper, J., Stringfellow, C., Elliot, R., Johnston, T. (1969). S,2(omеga-
aminoalkylamino)-ethyl dihydrogen phosphorothioates and related
compounds as potential antiradiation agents. Journal of Medical
Chemistry, 12, 236-243.
Piper, J. R., Rose, L. M., Johnson.T. P. Grenan, M. M. (1979). S-2-omega-
Diaminoalkyl dihydrogen phosphorothioates as antiradiation
agents.Journal of Medical Chemistry, 22, 613-639.
Pluquet, O., North, S., Bhoumik, A., Dimas, K., Ronai, Z., Hainaut, P.
(2003a).The cytoprotective aminothiol WR1065 activates p53 through
a non-genotoxic signaling pathway involving c-Jun N-terminal kinase.
Journal of Medical Chemistry, 278, 11879-11887.
Pluquet, O., North, S., Richard, M. J., Hainaut, P. (2003b).Activation of p53
by cytoprotective aminothiol WR1065: DNA-damage-independent
pathway and redox-dependent modulation of p53 DNA-binding ac-
tivity. Biochemical Pharmacology,65, 1129-1137.
Pomerantseva, M. D., Viklina, G. A. (1975). The effect of cystamine on the
outcome of dominant lethal mutations and reciprocal translocations in
sex cells of mice subjected to gamma-irradiation in different dos-
es.Genetika,10(7), 55-61.
Poplin, E. A., Lorusso, P., Lokich, J. J., Gullo, J. J., Leming, P. D., Schulz, J.
J., Veach, S. R., McCulloch, W., Baker, L., Schein, P. (1994). Ran-
domized clinical trial of mitomycin-C with or without pretreatment
with WR-2721 in patients with advanced colorectal cancer. Cancer
Chemotherapy and Pharmacology, 33, 415-419.
Purdie, J. W., Inhaber, E. R., Schneider, H., Labelle, J. L. (1983). Interaction
of cultured mammalian cells in the WR-2721 and its thiol WR-1065:
implications for mechanisms of radioprotection. International Journal
of Radiation Biology,43,517-527.
Ramdas, J., Warrier, R. P., Scher, C., Larussa, V. (2003). Effects of ami-
fostine on clonogenic mesenchymal progenitors and hematopoietic
progenitors exposed to radiation. Pediatric Hematology and Oncolo-
gy,25, 19-26.
Razgovorov, B. L., Saksonov, P. P., Antipov, V. V., Shashkov, V. S., Moro-
zov, V. S. (1971). The reactivity to some pharmacochemical drugs by
some of body shielding during total irradiation. In: Saksonov PP, Da-
vydov BI, eds, Radiobiological aspects of the reactivity of organism
during space flights. Problems of space biology. Vol. 14. Moscow:
Nauka. 1971. English translation accessed 1973 by NASA T T F–721.
Washington DC: National Aeronautics and Space Administration(pp.
163-175).
Razorenova, V. A., Shcherbova, E. N. (1961).On preventive use of cysteam-
ine and cysteinamine in acute radiation sickness.Meditsinskaia Radi-
ologiia,6, 11-14.
Robbers, H. Die pharmakologische Wirrung des Cystamins, einer
blutdrucksenkenden Substanz. (1937). Archiv fur Experimentelle Pa-
thologie und Pharmakologie, 185, 461.
Saksonov, P. P., Chernenko, G. T . (1959). Effect of mercamine on motor
function of the gastrointestinal tract.Farmakologiia i Toksikologiia, 22,
550-554.
Saksonov, P. P. (1975). Protection against radiation (biological, pharmaco-
logical, chemical, physical). In: Foundation of Space Biology and
Medicine.Vol. 3. Washington: NASA, (pp. 316-333).
Sapezhinskiĭ, I. I., Lozovskaia, E. L. (1992). The kinetics of the chemical
modification of DNA radiation transformations in multicomponent
systems.An analytical review.Radiobiologiia,32(2), 172-179.
 Journal of Radioprotection Research(2014) 15-36
Savoye, C., Swenberg, C., Hugot, S., Sy, D., Sabattier, R., Charlier, M.,
Spotheim-Maurizot,M. (1997). Thiol WR-1065 and disulfide WR-
33278, two metabolites of the drug ethyol (WR-2721) protect DNA
against neutron-induced strand. Interational Journal of Radiation Bi-
ology,71, 193-202.
Semenov, L. F., Larionov, L. F., Petrova, M. F., Pukhalskaia, E. C., Zeituni-
an, K. A. (1963). On the use of serototnin in the prevention of acute
radiation sickness in minkey.Meditsinskaia Radiologiia, (Moskow),
186, 58-62.
Shapira, R. D., Doherty, D. G., Burnett, W. T. (1957). Chemical protection
against ionizing radiation. III. Mercaptoalkylguanidines and related
isothiouronium compounds with protective activity. Radiation Re-
search, 7(1), 22-34.
Shashkov, V. S., Anashkin, O. D., Suvorov, N. N., Manaeva, I. A. (1971).
Efficacy of serotonin, mexamine, AET and cystamine with repeated
administration after γ-irradiation.Radiobiologiia, 11, 621-623.
Skrede, S. (1966). Effect of cystamine and cysteamine on the adenosine-tri-
phosphotase activity and oxidative phosphorilation of rat liver mito-
chondria.Biochemical Journal, 98, 702-704.
Smith, W. W., Budd, R. A., Cornfield, J. (1966). Estimation of radiation
dose-reduction factor for beta-mercaptoethylamine by endogenous
spleen colony counts.Radiation Research, 27, 363-368.
Sodicoff, M., Conger, A. D., T repper, P., Pratt, N. E. (1978).Short-term
radioprotective effects of WR-2721 on the rat parotid
glands.Radiation Research, 75(2), 317-326.
Soref, C. M., Hacker, T. A., Fahl, W. E. (2012). A new orally active, amino-
thiol radioprotector-free of nausea and hypotension side effects at its
highest radioprotective doses.InternationalJournal of Radiation On-
cology Biology Physics, 82, 701-707.
Storer, J. B., Coon J. M. (1950).Protective effects of para-amino-propio-
phenone against lethal doses of X-irradiation.Proceedingsof Socie-
tyfor Experimental Biology and Medicine, 74(1), 202-204.
Strelkov, R. B., Briantseva, L. A., Ziia, A. V. (1974). Possibility of practical
use of a gaseous hypoxic mixture for prevention of radiation inju-
ries.Radiobiologiia,14(2), 297-301.
Swenberg, C. E., Vaishnav, Y. N., Li, B., T sao, H., Mao, B., Geacintov, N.
E. (1997). Single-strand breaks in oligodeoxyribonucleotides induced
by fission neutrons and gamma radiation and measured by gel elec-
trophoresis: protective effects of aminothiols.Journal of Radiation Re-
search,38(4), 241-254.
T akahashi, I., Nagai, T., Miyaishi, K., Maehara, Y., Niibe, H. (1986). Clini-
cal study of radioprotective effects of amifostine (YM-08310, WR-
2771) on chronic radiation injury.International Journal of Radiation
Oncology Biology Physics,12, 935-938.
T eixeira, A., Morfim, M. P., de Cordova, C. A., Charão, C. C., de Lima, V.
R., Creczynski-Pasa, T. B. (2003). Melatonin protects against pro-
oxidant enzymes and reduces lipid peroxidation in distinct membranes
induced by the hydroxyl and ascorbyl radicals and by peroxyni-
trite.Journal of Pineal Research,35(4), 262-268.
T erekhov, A. V., Besedina, L. N., Zherebchenko, P. G., Znamenskiĭ, V. V.,
Suslikov, V. I., T itov, B. A. (1976). T oxicity and antiradiation activity
of aminopropyl aminoethyl thiophosphate.Radiobiologiia,16, 249-252.
T itov, A. V., Golubentsev, D. A., Mikhaĭlova, E. G. (1974).Species charac-
teristics of cystamine metabolism in mice and rats.Radiobiologiia, 14,
907-909.
T ravis, E. L., Fang, M. Z., Basic, I. (1988). Protection of mouse bone mar-
row by WR-2721 after fractionated irradiation. International Journal
of Radiation Oncology Biology Physics, 15, 377-382.
T remblay, M. E., Saint-Pierre, M., Bourhis, E., Lévesque, D., Rouillard, C.,
Cicchetti, F. (2006). Neuroprotective effects of cystamine in aged
parkinsonian mice.Neurobiology of Aging, 27, 862-870.
T rog, D., Bank, P., Wendt, T. G., Koscielny, S., Beleites, E. (1999). Daily
amifostine given concomitantly to chemoradiation in head and neck
cancer.A pilot study.Strahlentherapie und Oncologie, 175, 444-449.
T urrisi, A. T ., Kligerman, M. M., Glover, D. J., Glick, J. H., Norfleet, L.,
Gramkowski, M. (1983). I phasa of clinical investignation of WR-
2721.Radioprotectors and anticarcinogens. In: Nygaard O.F. (Ed),
NY, Academic Press (pp. 681-694).
Utley, J. F., Quinn, C. A., White, F. E., Seaver, N. A., Bloor, C. M. (1981).
Protection of normal tissue against late radiation injury by WR-
2721.Radiation Research, 85, 408-415.
Utley, J. F., Seaver, N., Newton, G. L., Fahey, R. C. (1984). Pharmacokinet-
ics of WR-1065 in mouse tissue following treatment with WR-
35
2721.International Journal of Radiation Oncology Biology Physics,
10, 1525-1528.
Vacek, A., Rоtkovska, D. (1964). On protective effect of 2,4-
dinitrophenol.International Journal ofRadiation Biology,8(3), 285-
289.
Vachon, A., Roman, V., Lecomte, C., Folcher, G., Fatôme, M., Braquet, P.,
Berleur, F. (1987). A radioprotector: cysteamine, inhibits oxygen
transport in lipidic membranes. International Journal of Radiation Bi-
ology,52, 847-852.
van Bekkum, D. W. (1956). The protective action of dithiocarbamates
against lethal effect of x-irradiation in mice.Acta Physiologica et
Pharmacologica Neerlandica,4(4), 508-523.
van den Brenk, H., Moore, R. (1959). Effect of high oxygen pressure on the
protective action of cystamine and 5-hydroxytryptamine in irradiated
rats. Nature,183(4674), 1530-1531.
van den Brenk, H., Eliot, K. (1958). 5-Hydroxytriptamine, radioprotective
action.Nature, 182(4648), 1506-1507.
van den Brenk, H., Jamieson, D. (1962). Studies of mechanisms of chemical
radiation protection in vivo. II. Effect of pressure oxygen on radio-
protection in vivo and its relation to “Oxygen poisoning”. Interna-
tional Journal ofRadiation Biology,4(4), 379-402.
van der Meer, C., van Bekkum D.W. (1959). The mechanism of radiation
protection by hystamine and other biological amines.International
Journal ofRadiation Biology,1(1), 5-12.
Van der Meer, C., van Bekkum, D. W. (1961). A study on the mechanism of
radiation protection by 5-hydroxytryptamine and trypta-
mine.International Journal ofRadiation Biology,4,105-110.
Vasilescu, D., Rix-Montel, M. A. (1980). Interaction of sulfur-containing
radioprotectors with DNA: a spectrophotometric study.Physiological
Chemistry and Physics,12(1), 51-55.
Vasin, M. V., Antipov, V. V. (1972). Radioprotective efficacy of cystamine
at repeat administration before -irradiation.Radiobiologiia,12, 924-
927.
Vasin, M. V., Antipov, V. V., Chernov, G. A., Abramov, M. M., Gavriliuk,
D. N., L'vova, T. S., Suvorov, N. N.(1997). The role of the vasocon-
strictor effect in realizing the radioprotective properties of indralin in-
experiments on dogs.Radiatsionnaia Biologiia Radioecologiia,37(1),
46-55.
Vasin, M. V., Antipov, V. V., Suvorov, N. N., Abramov, M. M.,Gorelova,
N. V. (1984). Characteristics of the role of the hydroxyl group in sero-
tonin in the pharmacological and antiradiation effect of serotonin. Ra-
diobiologiia,24(3), 411-414.
Vasin, M. V., Chernov Iu.N., Semenova, L. A. (1991). Antiradiation proper-
ties of radioprotectors, immunomodulators and agents affecting tissue
metabolism in fractionated irradiation.Radiobiologiia,31, 271-275.
Vasin, M. V., Davydov, B. I., Antipov, V. V. (1971). Comparative elimina-
tion of radioprotective and toxic effect of cystamine.Radiobiologiia,
11, 517-521.
Vasin, M. V., L’vova, T. S., Antipov, V. V., Dadydov, B. I. (1977). Radio-
protective efficacy of cystamine and mexamine on mice hematopoiet-
ic stem cells in vivo.Radiobiologiia, 17, 254-258.
Vasin, M. V., L’vova, T. S., Antipov, V. V., Dadydov, B. I. (1979). Radio-
sensitiveness of animals irradiated in altered gaseous medium. 1. The
influence of respiration by normobaric oxygen during irradiation on
radioresistance and radioprotective efficacy of radioprotectors. Radio-
biologiia,19, 712-715.
Vasin, M. V., Lebedeva, N. N. (1975). Effect of cystamine on the perfor-
mance of human operators.Kosmicheskaia Biologiia i Aviakosmiches-
kaia Meditsina,9, 54-57.
Vasin, M. V., Saksonov, P. P., Shashkov, V. S., Antipov, V. V. (1970b).
Relation between the anti-radiation activity of aminothiols (cysta-
mine), the dose of the preparation and the duration of its use under
various conditions of gamma-irradiation.Radiobiologiia,10,380–385.
Vasin, M. V., Saksonov, P. P., Shashkov, V. S. (1970a). Combined applica-
tion of radioprotective agents.Farmakologiia i Toksikologiia,33, 501-
504.
Vasin, M. V., Suvorov, N. N., Abramov, M. M., Gordeev, E. N. (1987).
Changes in the therapeutic spectrum with respect to the pharmacolog-
ical and radioprotective activity after O-alkylation of serotonin and
5(2-hydroxyethoxytryptamine).Radiobiologiia,27( 5),700–703.
Vasin, M. V., Ushakov, I. B., Koroleva, L. V., Antipov, V. V. (1999). The
role of cell hypoxia in the effect of radiation protec-
tors.RadiatsionnaiaBiologiiaRadioecologiia,39, 238-248.
36 Journal of Radioprotection Research(2014) 15-36
Vasin, M. V., Ushakov, I. B., Kovtun, V. Iu., Komarova, S. N., Semenova,
L. A. (2004). Effect of melatonin, ascorbic acid, and succinic acid on
the cumulative toxic effect of repeated treatment with gammafos
(amifostine).Bulletin of Experimental Biology and Medicine,137, 450-
452.
Vasin, M. V. (1977). Development and research of new effective agents of
pharmaco-chemical protection from damage ionizing radiation expo-
sure. Doctoral Dissertation. Moscow.
Vasin, M. V. (1986).Comparative characteristics of the modification of
radiosensitivity of mice and rats by a hypoxic hypox-
ia.Radiobiologiia,26, 563-565.
Verly, L. J., Sedlacek, R. (1983). Determination of the radioprotective ef-
fects of topical applications of MEA, WR-2721 and N-acetylcysteine
on murine skin.Radiation Research, 93, 175-183.
Vladimirov, V. G., Dzharakyan T. K. (1982). Radioprotective effects with
animals and human. Moscow:Atomizdat.
Vladimirov, V. G., Libikova, N. I. (1970).Cystamine effect to oxidative
phosphorilation in liver mitochondria.Farmakologiia i Toksikologiia,
33, 76-77.
Vladimirov, V. G., Strel'n ikov, Iu. E., Smirnova, S. M., Tarnapol'skaia, L. G.
(1993). The radioprotective efficacy and pharmacological properties
of S,beta-amidinoethylthiophosphate.Radiobiologiia,33, 116-121.
Vladimirov, V. G. (1968). A comparative study of the intracellular distribu-
tion of S35-cystamine in the liver and spleen of white mice and
rats.Radiobiologiia,8,258-261.
Ward, J. F. (1975). Radiation-induced strand breakage in DNA.Basic Life
Science, 5B, 471-472.
Wardman, P., Dennis, M. F., Stratford, M. R., White, J. (1992). Extra-
cellular: intracellular and subcellular concentration gradients of thiols.
International Journal of Radiation Oncology Biology Physics,22, 751-
754.
View publication stats
Washburn, L. C., Carlton, I. E., Hayes, R. L. (1974). Distribution of WR-
2721 in normal and malignant tissues of mice and rats bearing solid
tumors: dependence on tumor type, drug dose and species. Radiation
Research,59, 475-483.
Weiss, J. (1944). Radiochemistry of aqueus solutions.Nature,153, 748-750.
Woloschak, G. E., Paunesku, T ., Chang-Liu, C. M., Grdina, D, J. (1995).
Expression of thymidine kinase messenger RNA and a related tran-
script is modulated by radioprotector WR1065. Cancer Research,55,
4788-4792.
Xu, Y., Parmar, K., Du, F., Price, B. D., Sun, Y. (2011). The radioprotective
agent WR1065 protects cells from radiation damage by regulating the
activity of the Tip60 acetyltransferase.International Journal of Bio-
chemistry andMolecular Biology, 2, 295-302.
Yuhas, J. M., Proctor, J. O., Smith, L. H. (1973). Some pharmacologic ef-
fects of WR-2721: their role in toxicity and radioprotection. Radiation
Research,54,222-233.
Yuhas, J. M., Storer, J. B. (1969). Chemoprotection against three modes of
radiation death in the mice.International Journal of Radiation Biology,
15, 233-237.
Yuhas, J. M. (1970). Biological factors affecting the radioprotective effi-
ciency of S,2-(3-aminopropylamino) ethylphosphorothioic acid (WR-
2721) LD50/30 doses. Radiation Research, 44,621-628.
Yuhas, J. M. (1971). Biological factors affecting the radioprotective effi-
ciency of S,2-(3-aminopropylamino) ethylphosphorothioic acid (WR-
2721) LD50/7 doses. Radiation Research,7, 226-229.
Zheng, S., Newton, G. L., Ward, J. F., Fahey, R. C. (1992). Aerobic radio-
protection of pBR322 by thiols: effect of thiol net charge upon scav-
enging of hydroxyl radicals and repair of DNA radicals.Radiation Re-
search, 130(2), 183-193.
Zherebchenko, P. G., Suvorov, N. N. (1963). On the relation between the
radioprotective and vasoconstrictive action of indolylalkalyla-
mines.Radiobiologiia, 3(4), 595-602.