Combined chelation of lead (II) by deferasiroxand deferiprone in rats as biological model

F. Dahooee Balooch • S. J. Fatemi •

M. Iranmanesh

Received: 6 November 2013 / Accepted: 22 November 2013 / Published online: 6 December 2013

� Springer Science+Business Media New York 2013

Abstract In order to investigate the capability of

two chelators deferasirox (DFX or ICL670) and

deferiprone (L1) in removing lead from the body, the

present research was performed. Two does levels of 40

and 80 mg/kg body weight of lead (II) chloride was

given to rats as biological model for 45 days. After

45 days, some toxicity symptoms were observed in

rats such as loss of hair and weight, appearance of red

dots around eyes, weakness and irritability. After lead

application, chelation therapy with DFX and L1 as

mono and combined (DFX, L1 and DFX ? L1) was

done for 10 days. After chelation therapy, lead level in

different tissues reduced. The combined chelation

therapy results showed that these chelators are able to

remove lead from the body and toxicity symptoms

decreased. The combined therapy results (DFX ? L1)

show higher efficacy and lower toxicity compared to

single therapies.

Keywords Lead toxicity � Chelation therapy �Deferasirox � Deferiprone � Rats

Introduction

The excessive amount of pollutants such as heavy

metals in animal feed and feed stuffs are often due to

human actions, resulting from either agricultural,

industrial production, accidental or deliberate misuse

(Mohamed et al. 2009; Aboul-Enein et al. 2010; El-

Beltagi et al. 2010; Afify and El-Beltagi 2011). Lead

(Pb) is a heavy metal with no apparent biological

function. Lead is one of the toxic metals, it is

dangerous to most human body organ if exposure

exceed to tolerable levels. Lead can affect individuals

of any age, but it has a disproportionate effect on

children because their behavioral patterns place them

at higher risk for exposure to lead. Their bodies absorb

a larger percentage of the lead that they ingest and they

exhibit lead toxicity at lower level for exposure than

adults do. Accumulation of lead produces damaging

effects in the hematopoietical, hematic, renal and

gastrointestinal system (Ibrahim et al. 2012). The

chemical agents exposed in the work places affect

various organ and tissue systems, leading to chronic

diseases. Today, lead that is widely found in the nature

and has an increasing use with the industrial advance-

ments is used in printing, rubber, batteries, ceramics,

dye, porcelain manufacturing, accumulator industry

and as a gasoline additive. Especially, the workers of

these industrial branches, persons exposed to lead

environmentally develop disorders in the functions of

vital organs such as brain and kidneys related to the

toxic impact of lead (Hinc Y ilmaz et al. 2012).

F. D. Balooch � S. J. Fatemi (&)

Department of Chemistry, Faculty of Sciences, Shahid

Bahonar University of Kerman, 22 Bahman Blvd,

76169 Kerman, Iran

e-mail: fatemijam@yahoo.com

M. Iranmanesh

Department of Chemistry, Faculty of Sciences, Islamic

Azad University, Kerman Branch, Kerman, Iran

123

Biometals (2014) 27:89–95

DOI 10.1007/s10534-013-9689-0

Because of the size and charge similarities, Pb can

substitute Ca and can be deposited in the bone.

Especially, Children are susceptible to Pb-exposure

because the developing skeletal system requires high

calcium levels. Lead deposited in the bone is not

harmful, but may cause nephrotoxicity, neurotoxicity,

and hypertension (Kennish 1992). One of the major

concepts regarding the toxicity of heavy metals is

attributed to their ability to generate reactive oxygen

species (ROS), which cause oxidative stress (Ding

et al. 2001; Pande et al. 2001; Villeda-Hernandez et al.

2001; Hsu and Guo 2002) and consequently enhance

lipid peroxidation, decrease the saturated fatty acids

and increase the unsaturated fatty acid contents of

membranes (Malecka et al. 2001). Many authors

propose that the formation of free radicals is the most

important molecular mechanism of the lead toxicity

(Navarro-Moreno et al. 2009). Several studies have

shown that Pb can accumulate in the brain when its

concentration in the blood is elevated (Sansar et al.

2011). Only 1 h after intestinal absorption, lead

accumulates in bone, kidney, liver and other organs

(Navarro-Moreno et al. 2009). The toxicity of lead is

closely related to age, sex, and route of exposure level

of intake, solubility, metal oxidation state, retention

percentage, and duration of exposure, frequency of

intake, absorption rate, mechanisms and efficiency of

excretion. Lead has been associated with various

forms of cancer, nephrotoxicity, central nervous

system effects and cardiovascular diseases in human

(Ibrahim et al. 2012). In view of these considerations,

there is urgent need for development of a mechanism

for removal of toxic metal ions from the body.

Chelation therapy is the most effective way in

removing toxic element from the body. This kind of

treatment involves the use of chelating drugs that

coordinate to metal ion for the potentially fatal

conditions. These chelators increase the excretion

and subsequent depletion of transition metal in

biological system (Tubafard et al. 2010). In this

procedure, chelators are added to the blood through a

vein or administered orally in order to remove toxic

elements. Clinical investigations of some chelators in

removing various toxic metals in rats have been

previously published by Fatemi et al. (Amiri et al.

2007; Fatemi et al. 2007, 2009, 2012; Saljooghi and

Fatemi 2010; Iranmanesh et al. 2013). Deferasirox

{4-[3,5-bis(2-hydroxyphenyl)-1,2,4-triazol-1-yl]-ben-

zoic acid, ICL670, DFX, Fig 1a} is tridentate chelator

with high selectivity for Fe3? and indicates little

affinity for divalent ions such as Zn2? or Cu2?

(Saljooghi and Fatemi 2011). In vivo, this selectivity

is demonstrated by conserved plasma Zn and Cu levels

in patients taking DFX. DFX, in 2005, became the first

FDA approved oral alternative for treatment of iron

overload and subsequently was approved in the EU in

2006 (Yang et al. 2007). Its comparatively long half-

life before excretion allows once-daily dosage and

good overall patient compliance, as well as cost-

effectiveness. Deferasirox possesses a pFe3? value of

22.5 and can penetrate membranes easily and pos-

sesses good oral availability. Indeed, when orally

administered to hypertransfused rats, deferasirox pro-

motes the excretion of chelatable iron from hepato-

cellular iron stores four to five times more effectively

than desferrioxamine (Hershko et al. 2001). Deferi-

prone (1,2-dimethy1-3-hydroxypyride-4-one, L1,

Fig. 1a) was first reported as a potential orally active

iron chelator in 1984 (Hider et al. 1984) and demon-

strated to be active in man in 1987 (Kontoghiorghes

et al. 1987). L1 is water soluble and can be given orally

(Kontoghiorghes and Sheppard 1987). It possesses a

pFe3? value of 20.5 and its important property is its

ability to penetrate cells, coordinate iron, forming a

neutral complex, which is also capable of permeating

membranes (Glickstein et al. 2006). Recently suc-

cessful chelation therapy using both deferasirox and

deferiprone has been reported (Voskaridou et al.

2011). This kind of treatment by combining two

chelators is based on the assumption that various

chelating agents remove toxic metal from different

tissues and therefore better results are expected

(Tubafard et al. 2010). Results of this type of

combined therapy have been confirmed by Fatemi

NN

N

OH

OH

O

HO

N

OH

O

CH

CH

3

3 A B

Fig. 1 Chemical structures of deferasirox (a) and deferiprone

(b)

90 Biometals (2014) 27:89–95

123

et al. (2011, 2012) (Amiri et al. 2007; Tubafard and

Fatemi, 2008). The aim of present study was to

investigate the capability of DFX and L1 as chelating

agents in chelating and removing lead from the body.

Testing was performed by using a chronic poisoning

model on rats after lead application.

Experimental

Apparatus

Measurement of lead and iron in various tissues were

performed by a Varian flame atomic absorption

spectrometer (FAAS). A microwave oven, model

CEM MDS 200, was used to elimination the water

content present in organs and to facilitate digestion.

Also a Mettler analytical balance Model AE 160 was

used in this research.

Maintenance of the animals

Male Wistar rats were obtained from Kerman Neuro-

science Research center (Kerman, Iran). They kept in

animal house at this Center. The animals were kept

under a controlled light: dark (12:12 h) schedule at

23 ± 1 �C and humidity 50 %. The rats were divided

randomly to control and treated groups and were

maintained in well-cleaned sterilized cages. This

research was allowed by the ethics committee of

Shahid Bahonar University and Kerman Neuroscience

Research Center (Kerman, Iran).

Materials

Lead (II) chloride, L1 and other materials were

purchased from Merck Chemicals Co. and DFX was

purchased from Novartis Co. (Basel, Switzerland).

Methods

In our study, lead at two doses of 40 and 80 mg/kg

body weight was given to rats for 45 days.

In order to investigate side effects of different doses

of lead and evaluate abilities of chelators (DFX and

L1) in removing various doses of lead, we used two

doses of this metal (low and high doses). Animals were

classified as follows (Table 1): the first group (control

group) was given normal food and distilled water to

drink. Rats in this group were killed at the end of lead

administration (day 45). The concentrations of lead

and iron in tissues in control group were compared

with the groups that received lead and chelators. The

treated groups (low and high doses drinking of lead)

were given water containing 40 and 80 mg Pb2?/kg

body weight, respectively. The given dose volume for

rats was calculated based upon their weight. The

control group involved five animals and each of

treated groups had 25 animals. After 45 days, intox-

ication signs appeared and low and high doses groups

were divided to 5 sub-groups: 1. Before chelation

therapy 2. Without chelation therapy 3. Single therapy

with DFX 4. Single therapy with L1 and 5. Combined

therapy with DFX ? L1. In order to compare the lead

and iron concentrations in tissues, before and after

chelation therapy, one group was selected as before

chelation therapy group (vehicle). The rats, in this

group, were anesthetized with ether vapors and were

killed. Kidney, heart, liver, intestine and spleen

samples were weighed, dried and collected for deter-

mination of lead and iron content. In order to

investigate the effect of passing time in removing

lead from rat organs, without chelation therapy group

was selected. In order to investigate the capability of

two chelators (DFX and L1) to remove lead from the

body, chelation therapy was done. Chelators were

given immediately after lead application for 10 days.

Chelators DFX and L1 were given orally as single and

combined therapies. Doses of DFX and L1 were 140

and 300 mg/kg body weight, respectively. Lead

toxicity symptoms appeared in rats, were removed in

short term after drug application. Experiments were

done on 7-week-old Wistar male rats. There wasn’t

notable difference between the groups in the initial

body weight of the rats (mean 230 g), but at the end of

lead application test, those given lead in their diet had

remarkable weight loss (Table 2). Investigation of the

weights in this study indicates that dietary treatment

affected the food intake, whereby rats given normal

diet consumed more food than those given lead. At the

end of chelation therapy, animals were killed and

tissues were taken, dried and collected for determina-

tion of Pb and Fe concentrations. The samples were

put in an oven at 60 �C for 3 days. Then 1 g of each

samples were digested by 1 ml of HNO3. After

digestion, the solutions were vaporized with the

addition of 0.5 ml of H2O2 under the hood. After-

wards, the fragment was diluted with distilled water to

Biometals (2014) 27:89–95 91

123

10 ml volume. Determination of lead and iron in

samples were performed by atomic absorption spec-

trometry. The values are expressed as mean values (at

least three separate determinations) ± standard error

of mean (SEM). The data were subjected to statistical

analysis by Student’s t test; P \ 0.05 was considered

significant.

Result

Comparison of the animals weights indicates that dietary

treatment affects the food intake, whereby animals given

normal diet consumed more food than those given lead. A

significant difference between control and treated groups

was observed. Results of lead raising and iron reduction

in organs of treated groups were statistically different.

The lead accumulation in tissues at the 80 mg/kg dose

(high dose group) was greater than the group at 40 mg/kg

(low dose group). The toxicity symptoms appeared after

45 days of lead administration. Lead toxicity signs in

animals were observed as follows: appearance of red dots

around eyes, weakness, irritability, flaccid, decline

weight and loss of hair. Also, our results shown, as lead

concentration increased in tissues, iron concentration

decreased. The maximum amount of lead accumulation

was found in kidney followed by liver and spleen. After

the chelation therapy, the obtained results indicated that

present lead concentration in all tissues was significantly

reduced and the toxicity signs also decreased. There is

statistical difference between DFX and L1 in reducing the

amount of lead in various tissues. The t-test was applied

to the results assuming the certified values were the true

values. When we compared single therapy efficiencies of

chelators in this study, DFX was more effective in

decreasing lead level in all tissues, whereas, L1 was to be

more effective in kidneys. Comparison of single and

combined therapy shows combined group (DFX ? L1)

is more effective in reducing the lead concentration in all

tissues. The effects of chelators (DFX and L1) application

on lead concentration in the various tissues are shown in

Tables 3 and 4.

The difference between iron concentrations before

and after chelation therapies is remarkable. Iron

concentration was lowest in the group that had the

highest lead level, which is probably due to an

interference that could take place by lead through iron

uptake mechanism. Furthermore, iron concentration

after chelation therapy significantly decreased thus,

consumption of iron tablet is recommended to return

iron level to its normal state. Combination of deferasi-

rox ? deferiprone shows more efficiency in decreasing

iron level. The results of iron concentrations before and

after chelation therapies are shown in Tables 5 and 6. In

order to investigate the spontaneous elimination of lead

from the body by the biological system, one group was

treated without chelation therapy (Mohamed et al. 2009;

Navarro-Moreno et al. 2009). Removal of lead by the

biological system in this group is not noticeable.

Discussion

In order to investigate the abilities of DFX and L1

chelators in removing lead from the body, the

Table 1 Classification of animals

Control group

All

rats

Low and/or high lead

doses drinking

group

Before chelation therapy

Without chelation therapy

(vehicle)

Single therapy with DFX

(140 mg/kg body weight)

Single therapy with L1

(300 mg/kg body weight)

Combined therapy with DFX

(70 mg/kg body

weight) ? L1 (150 mg/kg

body weight)

Table 2 Body weights over 45 days for rats in different groups

Group Control Low dose drinking High dose drinking

Initial body weighta (g) 227 ± 6 (5) (1 day) 225 ± 4 (5) (1 day) 230 ± 6 (5) (1 day)

Final body weighta (g) 285 ± 5 (5) (45 day) 260 ± 7 (5) (47 day) 257 ± 4 (5) (47 day)

Values are mean for the number of observation in parenthesesa Mean of five determination ± standard deviation

92 Biometals (2014) 27:89–95

123

Table 3 Concentration of lead (mg/kg) in control and low dose groups

Group Control Before chelation

therapy

Without chelation

therapy

Chelation therapy

with DFX

Chelation therapy

with L1

Combined

chelation therapy

Heart 0.048 ± 0.008 0.510 ± 0.023 0.509 ± 0.019 0.175 ± 0.012 0.231 ± 0.014 0.108 ± 0.015

Kidney 0.032 ± 0.005 0.913 ± 0.023 0.911 ± 0.016 0.469 ± 0.023 0.421 ± 0.016 0.132 ± 0.014

Liver 0.037 ± 0.006 0.632 ± 0.018 0.631 ± 0.024 0.361 ± 0.017 0.473 ± 0.015 0.154 ± 0.016

Intestine 0.042 ± 0.011 0.378 ± 0.021 0.377 ± 0.017 0.316 ± 0.031 0.345 ± 0.025 0.215 ± 0.019

Spleen 0.052 ± 0.023 0.502 ± 0.012 0.498 ± 0.025 0.308 ± 0.018 0.387 ± 0.012 0.189 ± 0.014

The number of rats in each group was five; Results are represented as arithmetic mean ± SEM

Significant at P \ 0.05 when compared with control

Table 4 Concentration of lead (mg/kg) in control and high dose groups

Group Control Before chelation

therapy

Without chelation

therapy

Chelation therapy

with DFX

Chelation therapy

with L1

Combined

chelation therapy

Heart 0.048 ± 0.008 0.631 ± 0.017 0.629 ± 0.021 0.389 ± 0.019 0.434 ± 0.023 0.132 ± 0.012

Kidney 0.032 ± 0.005 1.182 ± 0.024 1.181 ± 0.018 0.674 ± 0.025 0.681 ± 0.019 0.183 ± 0.021

Liver 0.037 ± 0.006 0.853 ± 0.013 0.853 ± 0.021 0.281 ± 0.023 0.477 ± 0.018 0.203 ± 0.027

Intestine 0.042 ± 0.011 0.659 ± 0.022 0.658 ± 0.012 0.376 ± 0.027 0.411 ± 0.14 0.156 ± 0.23

Spleen 0.052 ± 0.023 0.787 ± 0.021 0.785 ± 0.018 0.456 ± 0.019 0.512 ± 0.011 0.153 ± 0.016

The number of rats in each group was five; Results are represented as arithmetic mean ± SEM

Significant at P \ 0.05 when compared with control

Table 5 Concentration of iron (mg/kg) in control and low dose groups

Group Control Before chelation

therapy

Without chelation

therapy

Chelation therapy

with DFX

Chelation therapy

with L1

Combined chelation

therapy

Heart 6.52 ± 0.33 6.32 ± 0.26 6.34 ± 0. 18 5.43 ± 0.32 5.57 ± 0. 11 5.21 ± 0.18

Kidney 4.85 ± 0.28 3.61 ± 0.24 3.61 ± 0.26 3.05 ± 0.23 3.21 ± 0.31 2.87 ± 0.23

Liver 6.53 ± 0.33 5.61 ± 0.29 5.64 ± 0.21 4.88 ± 0.24 5.12 ± 0.27 4.42 ± 0.27

Intestine 4.00 ± 0.24 3.86 ± 0.25 3.83 ± 0.27 3.51 ± 0.31 3.77 ± 0.21 3.11 ± 0.29

Spleen 4.36 ± 0.31 4.02 ± 0.28 4.05 ± 0.24 3.37 ± 0.29 3.67 ± 0.27 3.01 ± 0.35

The number of rats in each group was five; Results are represented as arithmetic mean ± SEM

Significant at P \ 0.05 when compared with control

Table 6 Concentration of iron (mg/kg) in control and high dose groups

Group Control Before chelation

therapy

Without chelation

therapy

Chelation therapy

with DFX

Chelation therapy

with L1

Combined chelation

therapy

Heart 6.52 ± 0.33 6.06 ± 0.23 6.08 ± 0. 27 5.23 ± 0.29 5.56 ± 0. 27 5.17 ± 0.33

Kidney 4.85 ± 0.28 3.75 ± 0.26 3.71 ± 0.31 3.31 ± 0.35 3.54 ± 0.29 3.15 ± 0.25

Liver 6.53 ± 0.33 5.73 ± 0.25 5.75 ± 0.28 5.37 ± 0.31 5.61 ± 0.24 5.10 ± 0.35

Intestine 4.00 ± 0.24 3.81 ± 0.31 3.80 ± 0.25 3.33 ± 0.22 3.59 ± 0.21 3.12 ± 0.28

Spleen 4.36 ± 0.31 4.01 ± 0.28 4.00 ± 0.19 3.67 ± 0.29 3.81 ± 0.34 3.43 ± 0.31

The number of rats in each group was five; Results are represented as arithmetic mean ± SEM

Significant at P \ 0.05 when compared with control

Biometals (2014) 27:89–95 93

123

distribution of lead was carried out and amount of

accumulation and toxic effects of lead on rats as

biological model was studied. In this study, a short-

term experimental model was used to speed up the

preliminary testing procedure. Lead is disharmonic

element with contexture of body. In contrast to other

vital elements such as zinc and selenium, tissues of

body identify lead as poison. Most of lead content

accumulate in kidney and liver. Chelation therapy is

one of the most effective ways to remove toxic metals

from the biological system. It has been reported that

the chelating agents having higher stability constants

with a metal in aqueous solution may also prove

successful in reducing the body burden of the metal

(Iranmanesh et al. 2013). Many studies have now

reported the high absorption/distribution, long-term

efficacy and safety of deferasirox and deferiprone in

removing some toxic metal ions and treating iron

overload in patients with b–thalassaemia major (Cap-

pellini 2008; Neufeld 2006; Hoffbrand et al. 2003).

After administration of chelators, the lead content

reduced, which shows that both of chelators (DFX, L1)

increases the removal of lead in tissues and also

toxicity symptoms were decreased. A comparison of

the results obtained from with and without chelation

therapies indicate that combined therapy (DFX ? L1)

enhances the removal of lead from rat organs consid-

erably and treat the side effects and general toxicity

symptoms caused by lead. Also, the toxicity and side

effects of combined therapy are lower than single

therapy. At present, combination therapy with deferi-

prone and desferrioxamine, that is highly selective for

iron(III) under biological conditions (pFe3? = 26.6),

is reported to be the most effective treatment for many

patients (Galanello et al. 2010). Desferrioxamine with

a higher pFe3? values acting as a sink. Presumably

deferasirox, also possessing a higher pFe3? values

than deferiprone, behaves in a similar manner.

Recently successful chelation therapy using both

deferasirox and deferiprone has been reported (Vosk-

aridou et al. 2011). In order for a chelating agent to

exert its pharmacological effect, a drug must be able to

reach the target sites at sufficient concentration. Each

of chelators (DFX and L1) has a different target tissue;

therefore, combination of them can help to remove

lead from various tissues effectively. The combined

therapy procedure is likely to increase metal excretion,

target specific metal tissues, minimize side effects (by

virtue of the use of lower doses) and improve

compliance. L1 by virtue of its bidentate nature and

the ability for iron(III) at neutral pH values, is highly

concentration dependent and at relatively low con-

centrations (\5 lM) the iron deferiprone complex will

donate iron to competing ligands (Devanur et al. 2008)

such as deferasirox. DFX, by virtue of its small size

and the ability to penetrate cells (Ma et al. 2012), has

capability of efficiency scavenging excess iron. By

considering of these characteristics, combination of

these chelators is more effective than single one in

removing of lead from rat organs. The important

finding that deferiprone leaves tissue iron at a level

close to normal is fundamental and would suggest that

the proposed use of this chelator will not be highly

toxic. The reason for this important observation is that

deferiprone is able to redistribute iron in mammals

(Evans et al. 2012). From the previous results obtained

by Fatemi et al. (2011, 2012) (Amiri et al. 2007;

Tubafard and Fatemi 2008) and our present results, it

can be concluded that two chelators (DFX ? L1) are

more effective as combined therapy than single

therapy in removal of lead from the body. Therefore,

DFX ? L1 combination seems to be a promising drug

of lead-mobilizing agent. This study might be effec-

tive for preliminary testing of the ability of chelating

agent in removing lead. Therefore after essential

preclinical experiments, this could be suggested for

human administration.

References

Aboul-Enein AM, AbouElella FN, Abdullah ES (2010) Moni-

toring of some organochlorines and organophosphorus

residues in imported and locally raised chicken and bovine

muscles in Egypt. J Appl Sci Res 6(6):600–608

Afify AMR, El-Beltagi HS (2011) Effect of the insecticide cy-

anophos on liver function in adult male rats. Fresen Envi-

ron Bull 20(4a):1084–1088

Amiri A, Fatemi SJ, Fatemi SN (2007) Removal of thallium by

combining desferrioxamine and deferiprone chelators in

rats. Biometals 20(2):159–163

Cappellini MD (2008) Long-term efficacy and safety of defer-

asirox. Blood Rev 22:35–41

Devanur LD, Neubert H, Hider RC (2008) The fenton activity of

iron(III) in the presence of deferiprone. J Pharm Sci

97:1454–1467

Ding Y, Gonick HC, Vaziri ND, Liang K, Wei L (2001) Lead-

induced hypertension. III. Increased hydroxyl radical pro-

duction. Am J Hypertens 14(2):169–173

El-Beltagi HS, Mohamed AA, Rashed MM (2010) Response of

antioxidative enzymes to cadmium stress in leaves and

94 Biometals (2014) 27:89–95

123

roots of radish (Raphanus sativus L.). Not Sci Biol

2(4):76–82

Evans RW, Kong X, Hider RC (2012) Iron mobilization from

transferrin by therapeutic iron chelating agents. Biochim

Biophys Acta 1820:282–290

Fatemi SJ, Amiri A, Bazargan MH, Tubafard S, Fatemi SN

(2007) Clinical evaluation of desferrioxamine (DFO) for

removal of thallium ions in rat. Int J Artif Org 30:902–905

Fatemi SJ, Tubafard S, Nadi B (2009) Evaluation of the effect of

cadmium on rat organs and investigation of diethyl car-

bamate as an oral drug in treatment of cadmium toxicity.

Med Chem Res 18(3):179–186

Fatemi SJ, Sh Saljooghi A, Dahooee BF, Iranmanesh M, Gol-

bafn MR (2011) Chelation of cadmium by combining

deferasirox and deferiprone in rats. Toxicol Ind Health

27(4):371–377

Fatemi SJ, Saljooghi AS, Balooch FD, Iranmanesh M, Golbafan

MR (2012) Removal of cadmium by combining deferasi-

rox and desferrioxamine chelators in rats. Toxicol Ind

Health 28(1):35–41

Galanello R, Agus A, Campus S, Danjou F, Giardina PJ, Grady

RW (2010) Combined iron chelation therapy. Ann NY

Acad Sci 1202:79–86

Glickstein H, BenEl R, Link G, Breuer W, Konijn AM, Hershko

C, Nick H, Cabantchik ZI (2006) Action of chelators in

iron-loaded cardiac cells: accessibility to intracellular labile

iron and functional consequences. Blood 108:3195–3203

Hershko C, Konijn AM, Nick HP, Breuer W, Cabantchik ZI,

Link G (2001) ICL670A: a new synthetic oral chelator:

evaluation in hypertransfused rats with selective radioiron

probes of hepatocellular and reticuloendothelial iron stores

and in iron-loaded rat heart cells in culture. Blood

97(4):1115–1122

Hider RC, Kontoghiorghes G, Silver J (1984) Pharmaceutical

compositions. UK patent 2,118,176A

Hinc Y ilmaz MD, Alper Keten MD, Emre Karacaoglu MD,

Engin Tutkun MD, Ramazan Akcan MD (2012) Analysis

of the hematological and biochemical parameters related to

lead intoxication. J Forensic Legal Med 19(8):452–454

Hoffbrand AV, Cohn A, Hershko C (2003) Role of deferiprone

in chelation therapy for transfusional iron overload. Blood

102:17–24

Hsu PC, Guo YL (2002) Antioxidant nutrients and lead toxicity.

Toxicology 180(1):33–44

Ibrahim NM, Eweis EA, El-Beltagi HS, Abdel-Mobdy YE

(2012) Effect of lead acetate toxicity on experimental male

albino rat. Asian Pac J Trop Biomed 2(1):41–46. doi:10.

1016/S2221-1691(11)60187-1

Iranmanesh M, Fatemi SJ, Ebrahimpour R, Dahooee FB (2013)

Chelation of chromium (VI) by combining deferasirox and

deferiprone in rats. Biometals 26(3):465–471

Kennish MJ (1992) Ecology of estuaries: anthropogenic effects.

CRC Press, Boca Raton. ISBN 9780849380419

Kontoghiorghes GJ, Sheppard L (1987) Simple synthesis of the

potent chelators 1-alkyl-3- hydroxy-2-mehylpyrid-4-ones.

Inorg Chim Acta 136:111–112

Kontoghiorghes GJ, Aldouri MA, Hoffbrand AV (1987)

Effective chelation of iron in beta thalassaemia with the

oral chelator 1,2-dimethyl-3-hydroxypyrid-4-one. Br Med

J 295:1509–1512

Ma Y, Zhou T, Kong X, Hider RC (2012) Chelating agents for

the treatment of systemic iron overload. Curr Med Chem

19:2816–2827. doi:10.2174/092986712800609724

Malecka A, Jarmuszkiewicz W, Tomaszewska B (2001) Anti-

oxidative defense to lead stress in sub cellular compart-

ments of pea root cells. Acta Biochim Pol 48:687–698

Mohamed AA, El-Beltagi HS, Rashed MM (2009) Cadmium

stress induced change in some hydrolytic enzymes, free

radical formation and ultrastructural disorders in radish

plant. Electron J Environ Agric Food Chem 8(10):967–983

Navarro-Moreno LG, Quintanar-Escorza MA, Gonzalez S,

Mondragon R, Cerbon-Solorzano J, Valdes J, Calderon-

Salinas JV (2009) Effects of lead intoxication on intercel-

lular junctions and biochemical alterations of the renal

proximal tubule cells. Toxicol In Vitro 23(7):1298–1304

Neufeld EJ (2006) Oral chelators deferasirox and deferiprone

for transfusional iron overload in thalassemia major: new

data, new questions. Blood 107(9):3436–3441

Pande M, Mehta A, Pant BP, Flora SJ (2001) Combined

administration of a chelating agent and an antioxidant in

the prevention and treatment of acute lead intoxication in

rats. Environ Toxicol Pharm 9(4):173–184

Saljooghi AS, Fatemi SJ (2010) Clinical evaluation of Defer-

asirox for removal of cadmium ions in rat. Biometals

23(4):707–712

Saljooghi AS, Fatemi SJ (2011) Removal of thallium by def-

erasirox in rats as biological model. J Appl Toxicol

31(2):139–143

Sansar W, Ahboucha S, Gamrani H (2011) Chronic lead

intoxication affects glial and neural systems and induces

hypoactivity in adult rat. Acta Histochem 113(6):601–607.

doi:10.1016/j.acthis.2010.06.005

Tubafard S, Fatemi SJ (2008) Chelation of bismuth by com-

bining desferrioxamine and deferiprone in rats. Toxicol Ind

Health 24(4):235–240

Tubafard S, Fatemi SJ, Saljooghi AS, Torkzadeh M (2010)

Removal of vanadium by combining desferrioxamine and

deferiprone chelators in rats. Med Chem Res 19:854–863.

doi:10.1007/s00044-009-9235-3

Villeda-Hernandez J, Barroso-Moguel R, Mendez-Armenta M,

Nava-Ruiz C, Huerta-Romero R, Rios C (2001) Enhanced

brain regional lipid peroxidation in developing rats

exposed to low level lead acetate. Brain Res Bull

55(2):247–251

Voskaridou E, Christoulas D, Terpos E (2011) Successful che-

lation therapy with the combination of deferasirox and

deferiprone in a patient with thalassaemia major and per-

sisting severe iron overload after single-agent chelation

therapies. Br J Haematol 154:654–656

Yang LP, Keam SJ, Keating GM (2007) Deferasirox: a review of

its use in the management of transfusional chronic iron

overload. Drugs 67(15):2211–2230

Biometals (2014) 27:89–95 95

123