Basic & Clinical Pharmacology & Toxicology, 2013, 112, 222228

Doi: 10.1111/bcpt.12019

Pharmacological Evaluation of a b-Hydroxyphosphonate

Analogue of L-Carnitine in Obese Zucker fa/fa Rats
Jorge Reyes-Esparza1, Brissa Mendoza-Rivera1, Ricardo De la Cruz-Cordero2, Jorge L. Rosado3, Miguel . Duarte-Vzquez2,
Mario G. Solis4, Odn Vite-Vallejo1 and Lourdes Rodrguez-Fragoso1
1
Facultad de Farmacia, Universidad Autnoma del Estado de Morelos, Cuernavaca, Morelos, Mxico, 2Nucitec, S. A. de C. V., Peuelas,
Queretaro, Mexico, 3CINDETEC A.C., Parque Industrial Quertaro, Quertaro, Mexico and 4Department of Pathology, General Hospital of
Tlahuac, Villa Centroamericana y del Caribe, Tlahuac, Mexico

(Received 10 May 2012; Accepted 17 September 2012)

Abstract: In this study, we evaluated the effect of an analogue of L-carnitine on parameters involved with Metabolic Syndrome
in obese Zucker rats. Twenty-four rats were treated for 5 weeks with L-carnitine (300 mg/kg) and its analogue at two concentrations (100 and 250 mg/kg) to assess their impact on glucose, triglycerides and cholesterol in liver and blood samples, as well as
the amount of liver glycogen. Liver slices were also analysed. The analogue reduced the levels of glucose, triglycerides and cholesterol in liver and the level of triglycerides in serum. At 100 mg/kg, the analogue proved more effective than L-carnitine in
improving the biochemical alterations present in liver. The amount of liver glycogen content was higher in obese animals treated
with both L-carnitine and the analogue. No changes on insulin and leptin were observed in animals treated. L-carnitine and its
analogue reduced the microvesicular fatty infiltration in liver. This study demonstrated that the analogue tested is more potent
and efficient than L-carnitine and improves the pharmacological profile of L-carnitine.

Metabolic syndrome, syndrome X or insulin resistance syndrome has been the focus of attention during the past 80 years
[1]. Its importance lies in its multiple, interrelated risk factors
for metabolic origin, which can lead to the development of
diseases such as atherosclerosis, type 2 diabetes mellitus,
dyslipidaemia, high blood pressure, elevated plasma glucose,
prothrombotic state and pro-inflammatory state [2]. Other factors associated with metabolic syndrome are abdominal obesity, insulin resistance and cardiovascular diseases (CVD) [3].
The main causes for the increase in the disease are obesity
and diabetes mellitus among populations with a sedentary life
pattern [4,5].
L-carnitine is synthesized by mammals as a result of the
synthesis of amino acids such as lysine or methionine and is
also obtained from the diet [6,7]. Carnitine is an essential factor in transporting the long-chain fatty acids (acyl CoA) from
the cytoplasm into the mitochondria where the beta-oxidation
takes place [8]. A large body of evidence suggests that carnitine and its derivatives acetyl-L-carnitine and propionyl-L-carnitine
enhance glucose utilization by stimulating the activity of pyruvate dehydrogenase complex [9], which is a key enzymatic
complex in glucose oxidation, because intramitochondrial
acetyl-CoA can be converted with carnitine into acetylL-carnitine (ALC) via the carnitine acetyltransferase that is
then transported out of the mitochondria [10]. L-Carnitine is
an amine key substrate in a family of carnitine acyltransferases, transferred reversibly between active units of acyl carnitine and co-enzyme A to preserve homoeostasis for a wide

Author for correspondence: Lourdes Rodrguez-Fragoso, Flavio Garca

No. 32, Presidentes Ejidales CP 04470, Mexico, D.F. (fax + 0052 777
3297089, e-mail [email protected]).

range of traffic that are crucial for acyl intermediary metabolism and cellular regulation [11].
Various analogues of L-carnitine have been synthesized to
study their mode of action and the structural features of their
binding sites [12,13]. The phosphorus analogues of naturally
occurring amino acids, which are produced by certain organisms, are of great interest in bioorganic and medicinal chemistry. The replacement of the carboxylic acid functional
group in biologically important molecules by phosphonic
acids continues to attract much interest in these fields [14
16]. Much of the progress in this area has been associated
with the phosphorus analogues of amino acids. These compounds have a tetrahedral configuration due to the presence
of the phosphorus atom, so they serve as stable analogues of
the unstable tetrahedral intermediate formed in enzymatic
processes. In a previous work [17], we evaluated the efficacy
and safety of two analogues of L-carnitine in insulin-resistant
Wistar rats with a high fructose diet. That study demonstrated that both analogues maintained the pharmacological
properties of L-carnitine and could improve the alterations
present in insulin-resistant treats; however and according to
the results, they varied in potency, effectiveness and toxicity.
The purpose of this study was to evaluate the effect of bhydroxy phosphonate analogue of L-carnitine under different
parameters involved with metabolic syndrome in obese Zucker rats.
Materials and Methods
Chemical synthesis of L-carnitine analogue. The starting material was
b-hydroxyphosphonate 4, obtained by modifying the method reported
by Ordez et al. [18]. The b-hydroxyphosphonate 4 was used in the
next reactions directly without isolation as a diastereomeric mixture.

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PHARMACOLOGICAL EVALUATION OF AN ANALOGUE OF L-CARNITINE

OH

OH

MeOH, 37-40 C
NBn2
rac-4

O
P(OMe)2

P(OMe)2 H2, Pd/C 10%

NH2
rac-5

CH3I, K2CO3
MeOH, 37-39 C
OH

P(OMe)2
I- +NMe

OH

-hydroxyphosphonate 3
(Analog of L-carnitine)

Crystallization

O
P(OMe)2

I- +NMe
rac-6

223

Hepatic triglycerides were also quantified through a colorimetric

method (Triglycerides SL; ELITech). Insulin and leptin were
quantified through ELISA kits (Rat/ Insulin and Rat/Leptin Kits;
Millipore, MO, USA).
Histopathological analysis. Liver tissue fragments were fixed in 10%
formaldehyde solution, dissolved in phosphate-saline buffer (pH 7.4),
dehydrated in alcohol and embedded in paraffin. Four-micrometre
paraffin sections were stained with haematoxylin and eosin (HE) and
subjected to histopathological examination.

Fig. 1. Representative scheme for the synthesis of the b-hiydroxyphosphonate analogue.

A diastereomeric mixture of compound 4 was hydrogenolized and

methylated, producing a racemic mixture of compound 6, 2-R and 2-S,
respectively, in proportion 85:15. The b-hiydroxyphosphonate 3 was
obtained diastereomerically pure (determined by 1H NMR and 31P
NMR) by fractional crystallization. Figure 1 shows the process
involved in the synthesis of L-carnitine analogue. L-carnitine and Lcarnitine analogue were obtained from Nucitec S.A de C.V.
(Queretaro, Mexico) and dissolved in sterile deionized water prior to
experimental use.
Animals and type of diets. Male Zucker rats (obese and lean) were
obtained at 5 weeks of age from Harlan Laboratories Inc., Mexico
City, Mexico. The lean animals were not genotyped and could have
been either +/+ or +/ for the leptin receptor deletion. The animals
were housed in a temperature- and humidity-controlled environment
and were allowed food (Standard Purina Chow Diet, Mexico) and
water ad libitum. Body-weight, glucose, triglycerides and cholesterol
were monitored throughout the study. The experiments were
conducted in accordance with the Guide for the Care and Use for
Laboratory Animals [19]. The Institutional Animal Care and Use
Committee from the Biotechnology Institute of the National
Autonomous University of Mexico approved all procedures.
Pharmacological treatments. After acclimatization, the animals were
divided into the following groups, each one consisting of six rats. The
lean Zucker rats received a control diet and water ad libitum and
comprised the control group. The obese Zucker rats were divided into
four groups. One group received a control diet and water ad libitum, a
second group received L-carnitine (250 mg/kg body-weight/day p.o. in
500 ll of water), a third group received the analogue at 100 mg/kg
body-weight/day p.o. in 500 ll of water, and the fourth group
received the analogue at 250 mg/kg body-weight/day p.o. in 500 ll of
water. All treatments were administered 5 days a week for the
duration of 5 weeks. L-carnitine and its analogue were obtained from
NUCITEC S.A. de C.V. and dissolved in sterile deionized water prior
to experimental use. The cholesterol, glucose and triglyceride serum
levels were monitored every 15 days. Food consumption and bodyweight were quantified every week. An oral glucose tolerance test
(3 g/kg) was performed after 5 weeks of treatment. After treatments,
animals were starved overnight and killed under light chloroform
anaesthesia. Liver tissue and blood samples were collected from each
animal and kept at 4C until further studies.
Biochemical analysis. Plasma from blood samples was collected by
centrifugation, and triglycerides, cholesterol and glucose levels were
quantified by colorimetric methods (Triglycerides SL, Cholesterol
PAP SL and Glucose PAP SL; ELITech, Morelos, Mexico). Glycogen
was assessed using the antrone method described by Fong et al. [20].

Statistical analysis. Results are expressed as means standard error

of the mean (S.E.M.). Baseline and post-treatment variables [peak
and area under the curve (AUC) glucose] were compared using a
paired t-test. AUC was calculated using the trapezoidal method.
Significant differences were detected by one-way analysis of
variance using the Graph Pad Instat program (Graph Pad Software
V2.03; Graph Pad Software Inc., San Diego, CA, USA). The Tukey
Kramer multiple comparison test was applied when significant
variations were found. Differences were considered significant at
p < 0.05.

Results
Body-weight gain and food consumption of obese Zucker rats.
The obese Zucker fa/fa rats were characterized by weight gain
and an increase in liver and serum triglyceride and cholesterol
levels throughout the period of the experiment (data not
shown). Body-weight gain and food consumption for both the
control and the obese groups are shown in fig. 2. After
5 weeks, obese Zucker fa/fa rats had gained 239 9.8 g
(fig. 2A), an increase in 84% compared with lean rats
(p < 0.05). At the end of treatment, animals treated with L-carnitine
and the analogue (100 and 250 mg/kg) had gained 245 18,
245 20 and 230 17 g in each case, 7788% when compared with lean rats (p < 0.05); no significant differences were
found between the experimental groups. All animals had a
gradual increase in body-weight, as can be seen in fig. 2A,
even though food amounts remained constant throughout
entire study and regardless of treatment.
Biochemical findings in obese Zucker rats.
Figure 3A shows the serum and liver glucose levels in obese
Zucker rats treated with carnitine and the analogue. No changes
in glucose levels were observed in serum from animals treated
with carnitine and analogue at 100 mg/kg. However, a significant increase (14%) was observed in animals receiving the analogue in 250 mg/kg doses. As we can see, obese animals
tended to increase their hepatic glucose levels, but this was not
statistically significant. Obese animals treated with L-carnitine
for 5 weeks did not show significant changes in liver glucose
levels when compared with obese rats. A reduction in liver glucose was nevertheless observed in obese animals treated with
100 mg/kg (30%) of the analogue (p < 0.05). Figure 3B shows
the results of the analysis of liver glycogen content. As shown,
the amount of liver glycogen changed in obese rats, resulting
in a 3.4 times increase when compared with the control group
(p < 0.05). Animals treated with L-carnitine showed an
increase in 48.3% in liver glycogen (p < 0.05), while animals

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224
A

Fig. 4. Effects of the analogue on glucose plasma concentrations in

obese Zucker rats after administering 3 g/kg glucose. Each bar represents the mean S.E.M. in experiments performed in duplicate. All
groups consisted of six animals. For abbreviations, see fig. 2.
*p < 0.05: statistically significantly different from the control group;
#
p < 0.05: statistically significantly different from obese rats.
&
p < 0.05: statistically significantly different from L-carnitine group.

Fig. 2. Effect of the analogue on weight gain and food intake. (A) Corporal weight was quantified every week during 5 weeks. (B) Food consumption was monitored every day during 5 weeks. All groups
consisted of six animals. A100 = analogue at 100 mg/kg; A250 = analogue at 250 mg/kg. *p < 0.05: statistically significantly different from
the control group; #p < 0.05: statistically significantly different from
obese rats.

250

mg/dl

250

Serum

Hepatic

200

200
150

150
100

100

#&

50

50

0
Lean

Obese

Obese + Car

Obese + A100 Obese + A250

18
16

*#

14

g/100 g

mg/dl/g of protein

*#

*#

12

10
8
6
4
2
0
Lean

Obese

Obese + Car

Obese + A100

Obese + A250

Fig. 3. Pharmacological effects of the analogue on glucose and glycogen from obese Zucker rats. (A) Hepatic and serum levels of glucose.
(B) Hepatic glycogen content. Each bar represents the mean S.E.M.
in experiments performed in duplicate. All groups consisted of six animals. For abbreviations, see fig. 2. *p < 0.05: statistically significantly
different from the control group; #p < 0.05: statistically significantly
different from obese rats. &p < 0.05: statistically significantly different
from L-carnitine group.

treated with 100 and 250 mg/kg of the analogue also showed a
49.3% and 48% increase in glycogen liver content when compared with obese rats (p < 0.05).
To know the effect of treatment on oral glucose tolerance,
we administered a bolus of 3 g/kg of glucose to animals and
quantified the plasma glucose concentrations. As can be seen
in fig. 4, obese animals had high concentrations of glucose,
remaining almost constant between 15 and 120 min., when
compared with the control group (AUC 54926 3547 versus
8751 987) (p < 0.05). On the other hand, animals treated
with L-carnitine showed a similar pattern to that observed in
the control group (AUC 6711 1890 versus 8751 987),
and there was a reduction in 88% in AUC when compared
with obese rats (p < 0.05). The patterns shown by analoguetreated animals, however, depended on the dose. Those treated
with 100 mg/kg had a pattern similar to that of the control
group (AUC 6892 2100 versus 8751 987) and a reduction
in 87.5% in AUC was observed in comparison with obese rats
(p < 0.05). The pattern of those treated with 250 mg/kg, on
the other hand, was very similar to that of the obese rats
(AUC 48791 4598 versus 54926 3547).
Figure 5 shows the insulin and leptin levels found in all
animals. As we can see, the insulin levels in obese animals
showed an important increase (20 times) (p < 0.05). Animals
treated with L-carnitine did not show any changes when compared with obese rats. Animals treated with 100 or 250 mg/kg
of the analogue tended to reduce slightly the insulin levels;
however, they were not significant. On the other hand, the leptin levels in obese animals showed also an important increase
(13 times) (p < 0.05); however, neither L-carnitine nor analogue modified the leptin levels.
Figure 6A shows that cholesterol serum levels increased by
62% in obese rats when compared with the control group.
However, obese animals treated with L-carnitine and 100 or
250 mg/kg of the analogue did not show significant changes

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225

Fig. 5. Effects of the analogue on insulin and leptin serum levels in

obese Zucker rats. Each bar represents the mean S.E.M. in experiments performed in duplicate. All groups consisted of six animals. For
abbreviations, see fig. 2. *p < 0.05: statistically significantly different
from the control group; #p < 0.05: statistically significantly different
from obese rats. &p < 0.05: statistically significantly different from
L-carnitine group.

Fig. 7. Effects of the analogue on hepatic and serum triglycerides levels in obese Zucker rats. Each bar represents the mean S.E.M. in
experiments performed in duplicate. All groups consisted of six animals. For abbreviations, see fig. 2. *p < 0.05: statistically significantly
different from the control group; #p < 0.05: statistically significantly
different from obese rats. &p < 0.05: statistically significantly different
from L-carnitine group.

Fig. 6. Effects of the analogue on hepatic and serum cholesterol levels

in obese Zucker rats. Each bar represents the mean S.E.M. in experiments performed in duplicate. All groups consisted of six animals.
For abbreviations, see fig. 2. *p < 0.05: statistically significantly different from the control group; #p < 0.05: statistically significantly different from obese rats. &p < 0.05: statistically significantly different
from L-carnitine group.

in cholesterol levels when compared to obese rats. Figure 6B

shows cholesterol levels increased by 66% in livers from
obese rats when compared with the control group (p < 0.05).
Animals treated with L-carnitine showed a reduction in 15% in
hepatic cholesterol; however, animals receiving 100 and
250 mg/kg of the analogue showed, respectively, a 60% and
50% reduction in hepatic cholesterol levels when compared
with obese rats (p < 0.05).
Hepatic and serum triglyceride levels are shown in fig. 7. A
five times increase in triglyceride serum levels was observed
for obese rats when compared to the control group (p < 0.05).
Obese animals treated for 5 weeks with L-carnitine showed a

16% reduction in triglyceride levels when compared to

untreated obese rats (p < 0.05) (fig. 7A). On the other hand,
obese animals treated with 100 or 250 mg/kg of the analogue
showed a reduction in triglyceride levels of 36.6% and 30%,
respectively, when compared to obese rats (p < 0.05).
Figure 7B shows triglyceride levels in liver from the control
and obese groups. Hepatic triglycerides showed a 2.5 times
increase in obese rats (p < 0.05). No reductions were observed
in animals treated with L-carnitine. However, obese animals
treated with 100 and 250 mg/kg of the analogue showed,
respectively, a 25% and 23% reduction in hepatic triglycerides
when compared with obese animals (p < 0.05).
Histopathological findings.
Figure 8 shows representative histological changes in the livers of the different groups. By week 5, obese Zucker rats
showed hepatocellular injury characterized by centrilobular,
microvesicular fatty infiltration, ballooning degeneration and
pleomorphic nuclei (fig. 8B). Obese rats treated with L-carnitine
showed a reduction in centrilobular, microvesicular fatty
infiltration and nuclear changes, along with cellular degeneration and the presence of inflammatory cells (fig. 8C). However, obese rats treated with 100 mg/kg of the analogue
showed significant improvement in the degree of fat accumulation in the liver and liver specimens displayed normal liver
histology and scattered inflammatory cells (fig. 8D). Obese
rats treated with 250 mg/kg of the analogue also showed a
reduction in fat accumulation in liver, but in less extension
(fig. 8E).

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226
A

Fig. 8. Effect of the analogue on liver slices histopathology. (A) Lean rat, (B) Obese rat, (C) Obese + carnitine, (D) Obese + A100 and (E)
Obese + A250. Liver slices were taken after 5 weeks of treatment. Arrows means microvesicular fatty infiltration; asterisk means the presence of
inflammatory cells. Haematoxilin and Eosin stain, magnification 9250.

Discussion
Metabolic syndrome has been recognized in medical literature
for over 80 years. It does not constitute a single illness.
Instead, it can be defined as a group of health problems
caused by genetic and environmental factors and the common,
fundamental pathogenic component of which is insulin resistance. These problems may occur simultaneously or one by
one, but their joint appearance in a single individual is significant given that these patients are more prone to CVD in general and to coronary disease in particular [21,22].
L-carnitine, a constituent of plasma and tissues, is biosynthesized from lysine and methionine in the liver [6]. It plays an
essential role in the transportation of free fatty acids into the
mitochondrial matrix for subsequent b-oxidation [6]. L-carnitine
occurs naturally in food items, especially red meat and fish.
The amounts ingested in a normal diet are, however, insufficient to meet the requirements of the entire body [8]. A biosynthetic pathway for L-carnitine has already been identified in
mammals, including human beings. Our study evaluated a
b-hydroxyphosphonate analogue of L-carnitine to identify its
pharmacological effects on certain parameters of metabolic
syndrome.
Genetically, obese (fa/fa) Zucker rats have proven being a
useful model for the study of metabolic syndrome because
they display several metabolic alterations, including obesity,
impaired glucose tolerance and liver steatosis [2325]. The
present study showed that obese rats suffered an increase in
triglycerides and cholesterol (in both serum and liver) as well
as liver glycogen and fatty liver and that L-carnitine treatment
improved liver cholesterol, serum TG, glucogen and oral glucose tolerance in this model. The present results demonstrated
that the b-hydroxyphosphonate analogue improved the phar-

macological effects compared with the original molecule

(L-carnitine) but had differential effects depending on the dose.
Doses of 250 mg/kg of the analogue only modified triglyceride levels in serum as well as triglycerides and cholesterol in
liver; the 100 mg/kg dose had a more powerful effect on triglyceride levels in serum while also reducing glucose, triglycerides and cholesterol in liver. Our results suggest that low
doses of the analogue had a better in vivo pharmacological
effect than those of L-carnitine. According to these results,
100 mg/kg of the analogue was more effective and potent than
L-carnitine when tested on obese Zucker rats. Analogue doses
of 250 mg/kg were less effective.
The role of carnitine in the regulation of fatty acid and carbohydrate metabolism is very complex. It is an essential cofactor
in the transfer of fatty acyl groups into the mitochondrial matrix,
where they undergo beta-oxidation [6,12]. Carnitine also plays a
role in the transfer of acetyl and other short acyl groups
from peroxisomes to mitochondria for further oxidation [13]. It
has been shown that dietary L-carnitine supplementation
decreased liver triglycerides as a result of an enhanced capacity
for b-oxidation [26]. As a result, it may help stimulate hepatic
glucose output and, therefore, may improve glucose status.
Recent studies suggest that L-carnitine has a beneficial impact
on the liver in cases of insulin resistance; it has modulatory
effects on the post-receptor level because it is able to enhance
tyrosine phosphorylation status [27]. We cannot discard the possibility that the b-hydroxyphosphonate analogue of L-carnitine
may act through the same mechanism. Results show that both
doses of the analogue reduced triglyceride levels in blood and
liver. Therefore, we can assume that this analogue of L-carnitine
might interfere with processes involved in b-oxidation and accumulation of lipotoxic metabolites that might contribute to mitochondrial dysfunction and insulin resistance.

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PHARMACOLOGICAL EVALUATION OF AN ANALOGUE OF L-CARNITINE

Obesity is the most significant single risk factor for the

development of fatty liver, both in children and in adults;
obesity is also predictive of the presence of fibrosis, potentially progressing into advanced liver disease [28]. From a
pathogenic point of view, insulin resistance plays a central
role in the accumulation of triglycerides inside the hepatocytes
and the initiation of the inflammatory cascade. Here, we found
that 100 mg/kg of the analogue reduces liver damage and
microvesicular fatty infiltration. Carnitine has also proven
effective in reducing plasma lipids and liver triglycerides in
the livers of rats chronically treated with ethanol [29]. However, the exact mechanism through which carnitine reduces
steatosis is still unknown. Recent findings suggest that the
hepatoprotective mechanism of carnitine involves dampening
of Kupffer cell TNF-a production [30]. Therefore, we cannot
discard the possibility that the analogue can ameliorate liver
injury through that mechanism.
Obese Zucker rats are considered representative of obesityassociated type diabetes in human beings. They are hyperlipidaemic, grossly obese, hyperinsulinaemic, insulin-resistant and
not prone to ketosis [31]. These animals are not always hyperglycaemic but exhibit an abnormal glucose tolerance similar to
that observed in human beings [32]. These rat glucose levels
are actually normal, or only slightly higher than normal.
Therefore, these animals are not the best models to study
effective treatments to control alterations of glucose homoeostasis. Our study did not find important changes in the glucose
serum, insulin serum and liver levels of obese rats. But the
animals displayed an abnormal plasma glucose concentration
during test tolerance, which was associated with impaired glucose tolerance; in that sense, our results agreed with previous
reports. Although L-carnitine did not modify the glucose and
insulin levels in serum and liver after 5 weeks of treatment,
the animals showed a normal rate of glucose tolerance. By
contrast, 100 mg/kg of the analogue reduced not only glucose
concentrations in serum and liver but also resulted in a normal glucose tolerance. This means that, in low doses, the bhydroxyphosphonate analogue is able to modify carbohydrate
metabolism better than L-carnitine.
Most mammalian cells store glycogen as a reserve for the
production of glucose 6-phosphate as a metabolic fuel for glycolysis. In liver, glycogen is mainly stored as a glucose reservoir for other tissues [33]. There is ample evidence human
beings with type 2 diabetes suffer from excess hepatic glucose
production and an abnormal hepatic glycogen metabolism.
Studies with Zucker rats have also shown dramatic findings
relate to regarding abnormal glycogen synthesis and storage
[31]. Our results agree with this. It has been found that ATP
production and glycogen synthesis increase with the addition
of L-carnitine to cultured hepatocytes from new born rats [34].
L-carnitine improves insulin-mediated glucose disposal either
in healthy individuals or in type 2 diabetic patients. There are
two possible mechanisms in the metabolic effect of carnitine:
the first is a regulation of acetyl and acyl cellular trafficking
for correctly meeting the energy demand; the second is a control action not only in the synthesis of key glycolytic and gluconeogenic enzymes, but also all glycogen-metabolizing

227

enzymes. Our results showed that the amount of glycogen in

liver was higher in L-carnitine-treated Zucker rats than in those
treated with the analogue. These results suggest that the analogue may produce the same effect on glycogen metabolism.
Zucker rats have displayed markedly elevated circulating
leptin levels and develop severe obesity with hyperphagia,
defective nonshivering thermogenesis and preferential deposition of energy in adipose tissue [3538]. It has been observed
that L-carnitine reduces obesity caused by high fat in C57BL/
6J mice [39]. Clinically, it has also been found that patients
treated with sibutramine/carnitine achieve significant weight
loss with a good safety profile [40]. L-carnitine has an antiobesity action, which could modulate lipid metabolism by stimulating lipolysis and oxidation, accompanied by corresponding
changes in gene expression and the expression and suppression of the expression of adipogenic genes [41]. Our study
found that Zucker rats reached twice the body-weight of their
lean counterparts; their weight gain was associated with an
accumulation in central fat and an increase in liver weight
(data not shown). Neither L-carnitine nor the analogue modified weight gain in Zucker rats. The discrepancy between our
results and previous work may be due to the animal model
used in the present study. Zucker rats present a mutation in
the leptin receptor, which is the molecular base of their characteristic phenotype (loss of satiety) [42]. L-carnitine and its
analogue may have an influence on fat and carbohydrate
metabolism but not on the central nervous system mechanism
of obesity and hyperphagia in Zucker rats. Our result showed
that although food consumption was constant during 5 weeks,
there was a progressive body-weight gain. Therefore, weight
gain was not dependent on food intake, but on the metabolism
and genetic alterations of the obese rats, as was corroborate by
the leptin levels.
This study demonstrates that the b-hydroxyphosphonate
analogue employed is more potent and efficient than L-carnitine
and improves the pharmacological properties of L-carnitine, as
well as it can improve the alterations present in fa/fa Zucker
rats. According to the present data, the analogues mechanisms of action could be similar to those reported for
L-carnitine.
Acknowledgements
We wish to thank to CONACYT Mexico for its financial
support (Project 153324 and 179848).
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2012 The Authors

Basic & Clinical Pharmacology & Toxicology 2012 Nordic Pharmacological Society