Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies
Aim: To study the influence of natural L-asparagine on the efficacy of cytostatic therapy for malignant tumors in experimental investigations. Materials and Methods: Female C57B1/6 mice weighing 18–20 g were selected for the experiments. Lewis’ lung carcinoma (LLC) and melanoma B16 cells were used i...
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irk-123456789-1386312018-06-20T03:08:26Z Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies Urazova, L.N. Kuznetsova, T.I. Boev, R.S. Burkova, V.N. Original contributions Aim: To study the influence of natural L-asparagine on the efficacy of cytostatic therapy for malignant tumors in experimental investigations. Materials and Methods: Female C57B1/6 mice weighing 18–20 g were selected for the experiments. Lewis’ lung carcinoma (LLC) and melanoma B16 cells were used in the study. Animals were inoculated with tumor cells intramuscularly. Solution of L-asparagine in a volume of 0.2 ml per mouse (in appropriate doses) was administered to the animals using gastric probe, daily, for 14 days. Cyclophosphane was administered intraperitoneally in total doses of 180 mg/kg and 90 mg/kg on days 3 and 7 after tumor implantation. The percentage of tumor growth inhibition was calculated and inhibition index and frequency of metastasis were assessed. Results: It has been shown that despite low activity of L-asparagine with regard to primary tumor, the level of metastasis inhibition is rather high (up to 91% depending on experimental model, therapy regimen and follow-up period). The analysis of previously obtained data and our studies indicate that L-asparagine derived from burdock (Arctium lappa) root has not only its own antimetastatic activity but it is also able to increase antimetastatic activity of cyclophosphane partially reducing toxic effect of cyclophosphane on the organism without decreasing its antitumor and antimetastatic activities. Conclusion: L-asparagine derived from burdock (Arctium lappa) root can be effective in the complex anticancer therapy with the use of appropriate chemotherapy doses and regimens. 2011 Article Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies / L.N. Urazova, T.I. Kuznetsova, R.S. Boev, V.N. Burkova // Experimental Oncology. — 2011. — Т. 33, № 2. — С. 90-93. — Бібліогр.: 18 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/138631 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Original contributions Original contributions Urazova, L.N. Kuznetsova, T.I. Boev, R.S. Burkova, V.N. Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies Experimental Oncology |
| description |
Aim: To study the influence of natural L-asparagine on the efficacy of cytostatic therapy for malignant tumors in experimental investigations. Materials and Methods: Female C57B1/6 mice weighing 18–20 g were selected for the experiments. Lewis’ lung carcinoma (LLC) and melanoma B16 cells were used in the study. Animals were inoculated with tumor cells intramuscularly. Solution of L-asparagine in a volume of 0.2 ml per mouse (in appropriate doses) was administered to the animals using gastric probe, daily, for 14 days. Cyclophosphane was administered intraperitoneally in total doses of 180 mg/kg and 90 mg/kg on days 3 and 7 after tumor implantation. The percentage of tumor growth inhibition was calculated and inhibition index and frequency of metastasis were assessed. Results: It has been shown that despite low activity of L-asparagine with regard to primary tumor, the level of metastasis inhibition is rather high (up to 91% depending on experimental model, therapy regimen and follow-up period). The analysis of previously obtained data and our studies indicate that L-asparagine derived from burdock (Arctium lappa) root has not only its own antimetastatic activity but it is also able to increase antimetastatic activity of cyclophosphane partially reducing toxic effect of cyclophosphane on the organism without decreasing its antitumor and antimetastatic activities. Conclusion: L-asparagine derived from burdock (Arctium lappa) root can be effective in the complex anticancer therapy with the use of appropriate chemotherapy doses and regimens. |
| format |
Article |
| author |
Urazova, L.N. Kuznetsova, T.I. Boev, R.S. Burkova, V.N. |
| author_facet |
Urazova, L.N. Kuznetsova, T.I. Boev, R.S. Burkova, V.N. |
| author_sort |
Urazova, L.N. |
| title |
Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies |
| title_short |
Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies |
| title_full |
Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies |
| title_fullStr |
Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies |
| title_full_unstemmed |
Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies |
| title_sort |
efficacy of natural l-asparagine in the complex therapy for malignant tumors in experimental studies |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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2011 |
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Original contributions |
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| citation_txt |
Efficacy of natural L-asparagine in the complex therapy for malignant tumors in experimental studies / L.N. Urazova, T.I. Kuznetsova, R.S. Boev, V.N. Burkova // Experimental Oncology. — 2011. — Т. 33, № 2. — С. 90-93. — Бібліогр.: 18 назв. — англ. |
| series |
Experimental Oncology |
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2025-07-10T06:14:35Z |
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2025-07-10T06:14:35Z |
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1837239458842279936 |
| fulltext |
90 Experimental Oncology 33, 90–93, 2011 (June)
EFFICACY OF NATURAL L-ASPARAGINE IN THE COMPLEX
THERAPY FOR MALIGNANT TUMORS IN EXPERIMENTAL STUDIES
L.N. Urazova1,*, T.I. Kuznetsova1, R.S. Boev2, V.N. Burkova3
1Cancer Research Institute, Siberian Branch of the Russian Academy of Medical Sciences, Tomsk, Russia
2Institute of Petroleum Chemistry, Siberian Branch of the Russian Academy of Medical Sciences, Tomsk, Russia
3Biolit-Ltd, Tomsk, Russia
Aim: To study the influence of natural L-asparagine on the efficacy of cytostatic therapy for malignant tumors in experimental inves-
tigations. Materials and Methods: Female C57B1/6 mice weighing 18–20 g were selected for the experiments. Lewis’ lung carci-
noma (LLC) and melanoma B16 cells were used in the study. Animals were inoculated with tumor cells intramuscularly. Solution
of L-asparagine in a volume of 0.2 ml per mouse (in appropriate doses) was administered to the animals using gastric probe, daily, for
14 days. Cyclophosphane was administered intraperitoneally in total doses of 180 mg/kg and 90 mg/kg on days 3 and 7 after tumor
implantation. The percentage of tumor growth inhibition was calculated and inhibition index and frequency of metastasis were assessed.
Results: It has been shown that despite low activity of L-asparagine with regard to primary tumor, the level of metastasis inhibition
is rather high (up to 91% depending on experimental model, therapy regimen and follow-up period). The analysis of previously obtained
data and our studies indicate that L-asparagine derived from burdock (Arctium lappa) root has not only its own antimetastatic activ-
ity but it is also able to increase antimetastatic activity of cyclophosphane partially reducing toxic effect of cyclophosphane on the
organism without decreasing its antitumor and antimetastatic activities. Conclusion: L-asparagine derived from burdock (Arctium
lappa) root can be effective in the complex anticancer therapy with the use of appropriate chemotherapy doses and regimens.
Key Words: Lewis lung carcinoma, melanoma B-16, L-asparagine, cyclophosphane.
Despite many advances in cancer treatment, tra-
ditional standard methods (surgery, radiation therapy
and chemotherapy) do not result in a complete cure
of cancer patients [1, 2]. Up to XVIII century, tumor
diseases were treated mainly with herbal extracts
and many empirically obtained data were confirmed
by up-to-date medicine [3, 4]. Herbs are “soft” drugs,
which combine a high biological activity with minimal
side effects [5]. Many studies have shown that many
herbal extracts, having moderate inhibitory effect
on tumor development, reduce toxic effect of cyto-
static agents on blood cells and increase the functional
activity of cells-effectors of immunity system as well
as increase chemotherapy efficacy and protect against
progression of metastases during surgery [6–9].
Studies of foreign authors and folk medicine ex-
perience in the use of extracts from burdock (Arctium
lappa) for cancer treatment are of great interest. Bur-
dock seeds contain arctiin and arctigenin, which have
anticancer effects. Cytotoxicity of these compounds
has been proven on cell cultures and inoculated tu-
mors [10–12]. Arctigenin shows cytotoxicity in 100%
of cases during oxygen-glucose deprivation in tu-
mor cells. The antiproliferative activity of glycosides
is caused by apoptosis induction [13, 14]. Extracts
from burdock leaves and roots are used in medicine
as agents having diuretic, diaphoretic, blood purifying
and anti-inflammatory effects. Antineoplastic effects
of ethanol and dichloromethane extracts of burdock
roots have been proved in experimental studies [15].
Apoptosis is an important component in the mecha-
nism of some antitumor agents. It is known that as-
paragine can be inductor of apoptosis in nervous tissue
[16]. We have found that the concentrated juice ob-
tained from fresh burdock roots has not only cytostatic
but also apoptosis-induced activity. The burdock root
contains up to 10.1% of free amino acids, maximum
amount of asparagine (6.4% in terms of raw mate-
rial and up to 50.0% in terms of the amount of amino
acids) [17].
Thus, it seems rather logical and interesting
to study antitumor activity of L-asparagine as one
of the main components included into the composition
of the studied agents derived from the burdock root.
MATERIALS AND METHODS
Female C57B1/6 mice weighing 18–20 g were
obtained from the Pharmacology Research Institute
in Tomsk, Russia. Research was conducted in ac-
cordance with the principles set out in the Guide for
the Care and Use of Laboratory Animals (Moscow
Breeding Nursery, Russian Acad. Med. Sci.).
Tumor strains. Lewis lung carcinoma (LLC) and
melanoma B16 cells were used in the study. Strains
obtained from the Russian Cancer Research Center
(Russia) are highly adequate models for studying
anti-tumor effect of biologic reaction modifiers of plant
origin. Animals were inoculated with tumor cells intra-
muscularly at the doses of 1×106 (LLC) and 5 ×106 (B16)
tumor cells per mouse.
Agents. Natural L-asparagine (Tomsk). The agent
was dissolved in warm distilled water. The obtained so-
lution in a volume of 0.2 ml per mouse (at doses of 100,
250 and 500 mg/kg was administered to the animals
Received: January 12, 2011.
*Correspondence: E-mail: url@oncology.tomsk.ru
Аbbreviations used: LLC — Lewis’s lung carcinoma; D-16 — mela-
noma B-16; CPH — cyclophosphane; IMI — index of metastasis
inhibition; TGI — percentage of tumor growth inhibition.
Exp Oncol 2011
33, 2, 90–93
Experimental Oncology 33, 90–93, 2011 (June) 91
using gastric probe in therapeutic regimen (48 h after
tumor implantation) daily, for 14 days.
Cyclophosphane. Cyclophosphane (Ministry
of Medical Industry, Russia) was used in mono-and
combined therapy for experimental tumors. Cyclo-
phosphane was administered intraperitoneally at the
total doses of 180 mg/kg and 90 mg/kg on days 3 and
7 after tumor implantation.
Changes in tumor growth were assessed by mea-
suring the tumor volume according to the Shrek for-
mula: V = A × B × C, where A, B, and C are orthogonally
related tumor sizes.
Percentage of tumor growth inhibition was cal-
culated by the formula: TGI% = [(Vc-Ve/Vc] × 100%,
where TGI% is a coefficient of tumor growth inhibition;
Vc is the average tumor volume in the control group
of animals; Ve is the average tumor volume in experi-
mental group of animals.
Index of metastasis inhibition. Metastatic spread
into lungs was evaluated on days 10 and 20 after tumor
inoculation. Mice were killed by cervical dislocation,
and their lungs examined. Index of metastatic inhibition
characterized the extent of metastatic involvement.
It was calculated by the formula: IMI = (Ac × Bc)-
(A × B)/(Ac × Bc) × 100%, where Ac and A are the
numbers of animals with metastases in the control
and experimental groups; Bc and B are the average
numbers of metastases in animals of the control and
experimental groups.
Frequency of tumor metastasis is the percent
of animals with metastases with respect to the overall
number of animals in the group.
Statistical analysis of results was carried out using
STATISTICA 6.0 software. Comparison between the
groups was made by nonparametric Mann — Whitney
test.
RESULTS AND DISCUSSION
Influence of the combination therapy with
L-asparagine and cyclophosphane on the growth
of Lewis lung carcinoma. The ability of amino acid
of L-asparagine to inhibit tumor growth and modulate
cytostatic therapy for experimental malignant tumors
was studied on mice with hematogenous metastasis
of Lewis’s lung carcinoma. This model is suitable
for studying anti-tumor and antimetastatic activities
of agents. Double injection of cyclophospane with
a 96-h interval is one of the standard chemotherapy
regimens in therapy of experimental tumors, in par-
ticular, LLC [18]. We injected cyclophosphane intra-
peritoneally in total doses of 180 mg/kg or 90 mg/
kg on days 3 and 7 after tumor implantation. Therapy
for LLC with L-asparagine at a dose of 250 mg/kg ad-
ministered alone resulted in insignificant antineoplastic
activity of this agent at late stages of oncogenesis
(up to 21.5% on day 20). Cyclophosphane at a dose
of 180 mg/kg led to statistically significant inhibition
of tumor growth ( p < 0.05) only at early stages of the
experiment (on day 7 and 10 with a maximum rate
of up to 61.8% on day 10 (Table 1).) Combination
of L-asparagine and CPH at a dose of 180 mg/kg de-
monstrated insignificant inhibition of primary tumor
growth as compared to monotherapy with CPH. Thus,
it has been shown that L-asparagine has not only in-
significant activity in monotherapy regimen, but it also
has insignificant ability to enhance therapeutic effect
of CPH with regard to primary tumor. The attempt
to reduce the CPH dose to 90 mg/kg resulted in inef-
fective therapy. Antitumor effect of the combination
therapy (TGI up to 21.9% on day 20) was compared
to that of using asparagine in monotherapy regimen.
CPH administered in the reduced dose demonstrated
insignificant antitumor effect (on day 20, TGI was
up to 14.9% at a dose of 90 mg/kg versus 44.7%
at a dose of 180 mg/kg).
Therapy with CPH at the doses of 180 mg/kg and
90 mg/kg resulted in respectively 26.7 and 10.1-fold
reductions in the number of metastases on day
20 in animals with LLC, which received the combina-
tion therapy with L-asparagine and CPH compared
to animals receiving no this combination therapy
(Table 2). Mice treated with CPH alone at the doses
of 180 mg/kg and 90 mg/kg demonstrated 18.8 and
2.7-fold reductions in the number of metastases,
respectively. It should be noted that IMI in animals
receiving therapy with asparagine in combination with
CPH at the doses of 180 mg/kg and 90 mg/kg was
rather high (98.5% and 90.1%, respectively). Me-
tastases in animals treated with asparagine and CPH
at a dose of 180 mg/kg were observed 2.6 times more
frequently than in animals receiving the reduced dose
of cyclophosphane. LLC therapy with L-asparagine
in combination with 180 mg/kg CPH led to insignifi-
cant weight loss of the liver and statistically significant
weight loss of the spleen (Table 3). The data obtained
can be indirect confirmation of reduction in toxic ef-
fect of CPH given in the combined therapy. We previ-
ously showed the reduction in mice spleen cellularity
after CPH injection that was partially decreased after
Table 1. Influence of L-asparagine (dose of 250 mg/kg) on the growth of Lewis lung carcinoma
Days
Groups of animals
Non
treted
n=15
Asparagine
n=15
CPH (I)
n=15
Asparagine +CPH (I)
n=15
CPH (II)
n=15
Asparagine+CPH (II)
n=15
V V TGI % р V TGI % р V TGI % р V TGI % р V TGI % р
7 1.2±0.2 1.5±0.1 - 0.24 0.6±0.1 49.58 0.03 0.5±0.1 55,46 0.02 1.2±0.2 1.68 0.73 1.3±0.2 - 0.88
10 2.9±0.1 3.0±0.1 - 0.19 1.13±0.1 60.35 0.0008 1.09±0.1 61.75 0.0006 3.0±0.2 - 0.6 2.6±0.2 7.72 0.37
13 4.0±0.9 4.4±0.3 - 0.67 1.4±0.2 64.4 0.01 2.5±0.2 37,37 0.18 4.7±0.5 - 0.56 3.8±0.3 4.04 0.81
17 5.7±1.2 4.7±0.5 18.71 0.39 3.2±0.3 43.36 0.042 3.8±0.2 34,27 0.2 5.9±0.7 - 0.81 5.1±0.3 11.36 0.83
20 7.0±0.9 5.5±0.8 21.45 0.25 3.9±0. 44.74 0.021 4.3±0.3 39.06 0.027 6.0±0.6 14.91 0.39 5.5±0.2 21.88 0.22
Notes: CPH (I) — dose оf 180 mg/kg; CPH (II) — dose of 90 mg/kg, V — tumor volume (cm3), TGI — tumor growth inhibition, n — number of mice.
92 Experimental Oncology 33, 90–93, 2011 (June)
administration of asparagine, i.e. toxic effect of CPH
on the organism was reduced.
Table 2. Influence of L-asparagine (dose of 250 mg/kg) on metastatic
spread of Lewis lung carcinoma
Groups of animals
Average number of metastases, IMI
10 day 20 day
M ± SE IMI % р M ± SE IMI % р
Non-treated, n=15 1±0,58 - - 101.3±8.7 - -
Asparagine, n=15 1±1 50.7 0.83 66.5±4.4 34.4 0.034
CPH (I) , n=15 0±0 100.0 0.19 5.4±2.2 94.7 0.024
Asparagine + CPH (I), n=15 0.3±0.3 83.5 0.38 3.8±1.8 98.5 0.024
CPH (II), n=15 0±0 100.0 0.19 37.6±12.0 62.9 0.024
Asparagine + CPH (II), n=15 0±0 100.0 0.19 10±2.0 90.1 0.025
Notes: CPH (I) — dose оf 180 mg/kg; CPH (II) — dose of 90 mg/kg, V — tu-
mor volume (cm3), TGI — tumor growth inhibition, n — number of mice; IMI —
index of metastasis inhibition.
Since the combined administration of CPH and L-
asparagine more markedly inhibits metastatic spread
nearly not influencing on the primary tumor, it can
be suggested that in this case the immune-mediated
inhibition of metastases but not the direct effect of CPH
on tumor cells takes place.
Influence of monotherapy with L-asparagine
on the growth of melanoma B-16. A statistically sig-
nificant inhibition of melanoma-16 growth on days 10 and
15 of the experiment was found in animals, which received
L-asparagine at a dose of 500 mg/kg (Table 4). Similar
results were obtained on days 8 and 15 in animals, which
received L-asparagine at a dose of 250 mg/kg. It should
be noted that on day 13 after tumor inoculation the rate
of tumor growth inhibition reduced to 4–7% depending
on the dose, then, on the day 15 the rate of tumor growth
inhibition slightly increased to 8–11%. Tumor volumes
of the control group animals were significantly greater than
those of the examined animals (see Table 4).
The study of the influence of natural L-asparagine de-
rived from burdock roots on metastatic spread of melano-
ma B-16 showed a high anti-metastatic effect of this agent
(Table 5). A statistically significant inhibition of metastasis
was found on days 13 and 15 of the experiment (up to 80%
and 91%, respectively). The phenomenon of inverse rela-
tionship between the dose of the agent and antimetastatic
effect clearly detectable on day 15 is of great interest. The
metastatic frequency at all stages of the tumor process
was 100% in all groups of animals.
Studies of activity of L-asparagine derived from
the burdock root extract using the model of LLC
showed insignificant antitumor and antimetastatic
activities of this agent administered alone. However,
the combination of asparagine and CPH at a dose
of 180 mg/kg demonstrated statistically significant
increase in metastasis inhibition (up to 99% on day
20 of the experiment). Similar tendency was observed
using another experimental model (melanoma B-16).
It was shown that asparagine used alone had insignifi-
cant antitumor activity, however, the rate of metastasis
inhibition was extremely high (up to 91% depending
on the therapy regimen and follow-up period). Thus,
the analysis of previous and our study results indicate
that natural L-asparagine derived from burdock roots
has not only its own anti-tumor activity but it is also
able to increase anti-tumor activity of CPH (at a total
therapeutic dose of 180 mg/kg) partially reducing its
toxic effect on the organism.
Table 4. Influence of L-asparagine on the growth of melanoma B-16
Days
Groups of animals
Non-
treated
n=15
L-asparagine (500 mg/kg),
n=15
L-asparagine (250 mg/kg),
n=15
V V TGI % р V TGI % р
8 0.5±0.1 0.4±0.1 33.3 0.1 0.4±0.07 35.2 0.031
10 2.0±0.2 1.5±0.1 17.2 0.05 1.7±0.1 14.2 0.14
13 3.5±0.2 3.0±0.1 6.6 0.07 3.3±0.1 4.3 0.5
15 4.9±0.1 4.3±0.2 11.2 0.03 4.5±0.1 7.9 0.02
Notes: V — tumor volume (cm3); ITG — inhibition of tumor growth.
Table 5. Influence of L-asparagine on metastatic spread of melanoma 16
Groups of animals
Average number of metastases and IMI
13 day 15 day
M ± SE IMI % p M ± SE IMI % p
Non-treated, n=15 2.9±1.1 29.5±4.5
L-asparagine (500 mg/kg), n=15 0.6±0.3 75.0 0.02 8.5±3.4 42.0 0.03
L-asparagine (250 mg/kg), n=15 0.4±0.2 80.0 0.004 1.7±0.7 91.0 0.05
Notes: IMI — index of metastasis inhibition.
Considering previously conducted studies, it can
be concluded that the agents derived from the burdock
root have detectable antitumor and antimetastatic
activities. Asparagine as one of the active components
of these agents, therefore, can be promising for cancer
therapy. Since these agents are used as a rule in the
complex therapy for malignant tumors, we consider
that further experimental studies on the influence
of L-asparagine on chemotherapy efficacy and surgery
are required.
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CPH(I), n=8 CPH(II), n=8 Asparagine +
CPH(II), n=8
Spleen 0.1±0.01 0.41±0.05 0.34±0.04
р=0.22
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р=0.22
0.21±0.0
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0.4±0.02
р=0.17
0.43±0.03
р=0.17
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1.24±0.11
р=0.6
1.00±0.14
р=0.35
1.28±0.06
р=0.05
1.32±0.06
р=0.6
Notes: CPH (I) — dose оf 180 mg/kg; CPH (II) — dose of 90 mg/kg, V — tumor volume (cm3), TGI — tumor growth inhibition, n- number of mice.
Experimental Oncology 33, 90–93, 2011 (June) 93
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