1Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 31 | Number 01 | Article 349875
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
JOURNAL VITAE
School of Pharmaceutical and
Food Sciences
ISSN 0121-4004 | ISSNe 2145-2660
University of Antioquia
Medellin, Colombia
Filliations
1 Department of Biology, College of
Science, University of Baghdad,
Baghdad-Iraq
*Corresponding
Shaymaa Fadhel Abbas Albaayit
shaymaa_albaayit@yahoo.com
Received: 01 June 2022
Accepted: 15 February 2023
Published: 27 February 2024
Enzyme Inhibitory and Anti-cancer
Properties of Moringa peregrina
Propiedades inhibidoras Enzimáticas y Anticancerígenas de
la Moringa peregrina
Shaymaa Fadhel Abbas Albaayit 1
ABSTRACT
Background: Moringa peregrina is widely used in the traditional medicine of the Arabian
Peninsula to treat various ailments, because it has many pharmacologically active components
with several therapeutic effects. Objective: This study aimed to investigate the inhibitory
effect of Moringa peregrina seed ethanolic extract (MPSE) against key enzymes involved in
human pathologies, such as angiogenesis (thymidine phosphorylase), diabetes (α-glucosidase),
and idiopathic intracranial hypertension (carbonic anhydrase). In addition, the anticancer
properties were tested against the SH-SY5Y (human neuroblastoma). Results: MPSE extract
significantly inhibited α-glucosidase, thymidine phosphorylase, and carbonic anhydrase with
half-maximal inhibitory concentrations (IC 50 ) values of 303.1 ± 1.3, 471.30 ± 0.3, and 271.30 ±
5.1 μg/mL, respectively. Furthermore, the antiproliferative effect of the MPSE was observed on
the SH-SY5Y cancer cell line with IC 50 values of 55.1 μg/mL. Conclusions: MPSE has interesting
inhibitory capacities against key enzymes and human neuroblastoma cancer cell line.
Keywords: α-glucosidase, carbonic anhydrase, thymidine phosphorylase, neuroblastoma,
Moringa peregrina.
ORIGINAL ARTICLE
Published 27 February 2024
Doi: https://doi.org/10.17533/udea.vitae.v31n1a349875
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
JOURNAL VITAE
School of Pharmaceutical and
Food Sciences
ISSN 0121-4004 | ISSNe 2145-2660
University of Antioquia
Medellin, Colombia
Filliations
1 Department of Biology, College of
Science, University of Baghdad,
Baghdad-Iraq
*Corresponding
Shaymaa Fadhel Abbas Albaayit
shaymaa_albaayit@yahoo.com
Received: 01 June 2022
Accepted: 15 February 2023
Published: 27 February 2024
Enzyme Inhibitory and Anti-cancer
Properties of Moringa peregrina
Propiedades inhibidoras Enzimáticas y Anticancerígenas de
la Moringa peregrina
Shaymaa Fadhel Abbas Albaayit 1
ABSTRACT
Background: Moringa peregrina is widely used in the traditional medicine of the Arabian
Peninsula to treat various ailments, because it has many pharmacologically active components
with several therapeutic effects. Objective: This study aimed to investigate the inhibitory
effect of Moringa peregrina seed ethanolic extract (MPSE) against key enzymes involved in
human pathologies, such as angiogenesis (thymidine phosphorylase), diabetes (α-glucosidase),
and idiopathic intracranial hypertension (carbonic anhydrase). In addition, the anticancer
properties were tested against the SH-SY5Y (human neuroblastoma). Results: MPSE extract
significantly inhibited α-glucosidase, thymidine phosphorylase, and carbonic anhydrase with
half-maximal inhibitory concentrations (IC 50 ) values of 303.1 ± 1.3, 471.30 ± 0.3, and 271.30 ±
5.1 μg/mL, respectively. Furthermore, the antiproliferative effect of the MPSE was observed on
the SH-SY5Y cancer cell line with IC 50 values of 55.1 μg/mL. Conclusions: MPSE has interesting
inhibitory capacities against key enzymes and human neuroblastoma cancer cell line.
Keywords: α-glucosidase, carbonic anhydrase, thymidine phosphorylase, neuroblastoma,
Moringa peregrina.
ORIGINAL ARTICLE
Published 27 February 2024
Doi: https://doi.org/10.17533/udea.vitae.v31n1a349875
2Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 31 | Number 01 | Article 349875Shaymaa Fadhel Abbas Albaayit
RESUMEN
Antecedentes: La Moringa peregrina se utiliza ampliamente en la medicina tradicional de la Península Arábiga para tratar diversas
dolencias, ya que posee numerosos componentes farmacológicamente activos con varios efectos terapéuticos. Objetivo: Este
estudio tenía como objetivo investigar el efecto inhibidor del extracto etanólico de semillas de Moringa peregrina (MPSE) frente
a enzimas clave implicadas en patologías humanas, como la angiogénesis (timidina fosforilasa), la diabetes (α-glucosidasa) y la
hipertensión intracraneal idiopática (anhidrasa carbónica). Además, se comprobaron las propiedades anticancerígenas frente
al SH-SY5Y (neuroblastoma humano). Resultados: El extracto de MPSE inhibió significativamente la α-glucosidasa, la timidina
fosforilasa y la anhidrasa carbónica con concentraciones inhibitorias semimáximas (IC50) de 303,1 ± 1,3, 471,30 ± 0,3 y 271,30
± 5,1 μg/mL, respectivamente. Además, se observó el efecto antiproliferativo del MPSE en la línea celular del cáncer SH-SY5Y
con valores de IC50 de 55,1 μg/mL. Conclusiones: MPSE posee interesantes capacidades inhibitorias frente a enzimas clave y
línea celular de neuroblastoma canceroso humano.
Palabras clave: α-glucosidasa, anhidrasa carbónica, timidina fosforilasa, neuroblastoma, Moringa peregrina.
INTRODUCTION
In metabolism, various enzymes act as biocatalysts
that interact with foreign substances. In some
cases, enzymes are known to act as drug targets
(1) for human diseases. Treating these diseases with
synthetic enzyme inhibitors often fails due to toxicity
and side effects. Therefore, it is necessary to search
for non-cytotoxic and effective enzyme inhibitors
agents from natural sources (2).
Traditional herbal medicine has extensively used
different plant parts to treat various disorders such
as cardiovascular, gastrointestinal, diabetes, wound
healing, and microbial infections. (3-8). Moringa
peregrina (Forssk) Fiori is a well-known herbal
plant, and the wide range of pharmacological
proper ties of its seeds have been repor ted,
including immunomodulatory effect, antioxidant,
antimicrobial activity, analgesic, and cytotoxic
effects against several cancer cell lines (9,10),
which can be exploited in the development of
drug candidates. The seeds of M. peregrina are
traditionally used to control diabetes, wounds,
gastrointestinal diseases, anti-inflammatory, and
anticancer (11, 12). Some active phytochemicals in
M. peregrina (triterpenoids, tocopherols, flavonoids,
isothiocyanate, quercetin, and phenolics) have been
well-reported to enhance the immune system to
fight against cancer cells (10, 13-15).
In type II diabetes mellitus, cells resist insulin,
and glucose is not properly utilized, leading to
hyperglycemia. The α-glucosidase inhibitors
have been proposed as a promising approach to
treating diabetes. A drug that inhibits α-glucosidase
activity prevents disaccharides’ breakdown into
monosaccharides, lowering blood glucose levels.
This approach causes a decrease in glucose
absorption from the gastrointestinal tract, preventing
the rise in glucose levels after food consumption.
Therefore, researchers are looking for plant-based
products with anti-α-glucosidase properties (16).
Thymidine phosphor ylase (TP) is an enzyme
responsible for the initiation of angiogenesis. Hyper
activation of thymidine phosphorylase is involved in
tumor development by promoting endothelial cell
migration and the release of various angiogenic
fac tors from malignant and stromal cells in
the tumor microenvironment site and helps
in the evasion of apoptosis and immune cells.
Therefore, it is necessary to discover therapeutic
agents that can stop angiogenesis (17).
Carbonic anhydrase is a well-known drug-target
enzyme responsible for physiological processes
related to respiration and the transport of CO2 /
bicarbonate between metabolizing tissues and
the lungs. Carbonic anhydrase is involved in
biosynthetic reactions such as bone resorption,
calcification, gluconeogenesis, lipogenesis, and
ureagenesis. Carbonic anhydrase enables tumor
growth compared to normal tissues (18).
The rapid development of resistance in cancer
cells to prevailing anticancer drugs and ineffective
surgical or radiotherapy had often failed to reduce
the chances of recurrence and ultimately led to the
progression of cancer to the metastatic stage (19,
20). In the recent decades, researchers have paid
attention to natural products as active compounds
isolated from plants that are considered safe and
have pharmacological anticancer effects (21, 22).
The current research was the first-time report on
the effect of M. peregrina seed extract on the
enzyme inhibition potential for α-glucosidase,
thymidine phosphorylase, and carbonic anhydrase.
RESUMEN
Antecedentes: La Moringa peregrina se utiliza ampliamente en la medicina tradicional de la Península Arábiga para tratar diversas
dolencias, ya que posee numerosos componentes farmacológicamente activos con varios efectos terapéuticos. Objetivo: Este
estudio tenía como objetivo investigar el efecto inhibidor del extracto etanólico de semillas de Moringa peregrina (MPSE) frente
a enzimas clave implicadas en patologías humanas, como la angiogénesis (timidina fosforilasa), la diabetes (α-glucosidasa) y la
hipertensión intracraneal idiopática (anhidrasa carbónica). Además, se comprobaron las propiedades anticancerígenas frente
al SH-SY5Y (neuroblastoma humano). Resultados: El extracto de MPSE inhibió significativamente la α-glucosidasa, la timidina
fosforilasa y la anhidrasa carbónica con concentraciones inhibitorias semimáximas (IC50) de 303,1 ± 1,3, 471,30 ± 0,3 y 271,30
± 5,1 μg/mL, respectivamente. Además, se observó el efecto antiproliferativo del MPSE en la línea celular del cáncer SH-SY5Y
con valores de IC50 de 55,1 μg/mL. Conclusiones: MPSE posee interesantes capacidades inhibitorias frente a enzimas clave y
línea celular de neuroblastoma canceroso humano.
Palabras clave: α-glucosidasa, anhidrasa carbónica, timidina fosforilasa, neuroblastoma, Moringa peregrina.
INTRODUCTION
In metabolism, various enzymes act as biocatalysts
that interact with foreign substances. In some
cases, enzymes are known to act as drug targets
(1) for human diseases. Treating these diseases with
synthetic enzyme inhibitors often fails due to toxicity
and side effects. Therefore, it is necessary to search
for non-cytotoxic and effective enzyme inhibitors
agents from natural sources (2).
Traditional herbal medicine has extensively used
different plant parts to treat various disorders such
as cardiovascular, gastrointestinal, diabetes, wound
healing, and microbial infections. (3-8). Moringa
peregrina (Forssk) Fiori is a well-known herbal
plant, and the wide range of pharmacological
proper ties of its seeds have been repor ted,
including immunomodulatory effect, antioxidant,
antimicrobial activity, analgesic, and cytotoxic
effects against several cancer cell lines (9,10),
which can be exploited in the development of
drug candidates. The seeds of M. peregrina are
traditionally used to control diabetes, wounds,
gastrointestinal diseases, anti-inflammatory, and
anticancer (11, 12). Some active phytochemicals in
M. peregrina (triterpenoids, tocopherols, flavonoids,
isothiocyanate, quercetin, and phenolics) have been
well-reported to enhance the immune system to
fight against cancer cells (10, 13-15).
In type II diabetes mellitus, cells resist insulin,
and glucose is not properly utilized, leading to
hyperglycemia. The α-glucosidase inhibitors
have been proposed as a promising approach to
treating diabetes. A drug that inhibits α-glucosidase
activity prevents disaccharides’ breakdown into
monosaccharides, lowering blood glucose levels.
This approach causes a decrease in glucose
absorption from the gastrointestinal tract, preventing
the rise in glucose levels after food consumption.
Therefore, researchers are looking for plant-based
products with anti-α-glucosidase properties (16).
Thymidine phosphor ylase (TP) is an enzyme
responsible for the initiation of angiogenesis. Hyper
activation of thymidine phosphorylase is involved in
tumor development by promoting endothelial cell
migration and the release of various angiogenic
fac tors from malignant and stromal cells in
the tumor microenvironment site and helps
in the evasion of apoptosis and immune cells.
Therefore, it is necessary to discover therapeutic
agents that can stop angiogenesis (17).
Carbonic anhydrase is a well-known drug-target
enzyme responsible for physiological processes
related to respiration and the transport of CO2 /
bicarbonate between metabolizing tissues and
the lungs. Carbonic anhydrase is involved in
biosynthetic reactions such as bone resorption,
calcification, gluconeogenesis, lipogenesis, and
ureagenesis. Carbonic anhydrase enables tumor
growth compared to normal tissues (18).
The rapid development of resistance in cancer
cells to prevailing anticancer drugs and ineffective
surgical or radiotherapy had often failed to reduce
the chances of recurrence and ultimately led to the
progression of cancer to the metastatic stage (19,
20). In the recent decades, researchers have paid
attention to natural products as active compounds
isolated from plants that are considered safe and
have pharmacological anticancer effects (21, 22).
The current research was the first-time report on
the effect of M. peregrina seed extract on the
enzyme inhibition potential for α-glucosidase,
thymidine phosphorylase, and carbonic anhydrase.
3Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 31 | Number 01 | Article 349875
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
Fur thermore, the anticancer potential of M.
peregrina seed extract was investigated in the
human neuroblastoma cell line SH-SY5Y.
MATERIALS AND METHODS
Plant Preparation
M. peregrina seeds were submitted and authenticated
by Dr. Maha Kordofani (Resident Botanist) at the
Department of Botany, University of Khartoum. M.
peregrina seeds were kept at room temperature
until they were dried and ground to powder. 100 g
of powder was mixed s with 500 mL of ethanol and
left for 5 days for proper mixing. The macerates were
filtered through a muslin cloth and collected; then,
rotary evaporated to obtain the crude M. peregrina
seed extract (MPSE) (23).
Alpha-Glucosidase inhibition assay
20 μL of MPSE prepared in different concentrations
(0.125-1 mg/mL) were added to a 96-well plate.
Along with the test extract, 20 μL of enzyme solution
(0.2 U/mL) and 135 μL of 0.1 M phosphate buffered
saline (PBS) buffer (pH 6.8) were added to each well.
The plate was then incubated at 37 °C for 15 min.
After incubation, the absorbance was measured
before the substrate addition. The substrate (25 μL)
4-nitrophenyl-α-D-glucopyranoside (PNP-G)
(0.7 mM) was then added, and the change in
absorbance was measured for 30 min at 400 nm in
a spectrophotometer (Spectra Max, MD, USA) (24).
Acarbose served as the positive (drug) control.
Thymidine phosphorylase (TP) inhibition assay
We followed the protocol of Javaid et al. (25). Briefly,
10 μL of MPSE at different concentrations (0.125-1
mg/mL) were incubated with 20 μL of the enzyme
(0.058 unit/well) along with 150 μL of potassium
phosphate buffer (pH 7.0, 50 mM) for 10 min at 30°C.
After 10 min, 20 μL (1.5 mM) substrate (thymidine,
kmax; 265 nm) was added. Soon after, absorbance
was recorded at 290 nm using an ELISA plate reader
and continued for 10 min to monitor the change in
absorbance. 7-Deazaxanthine served as the positive
(drug) control.
Carbonic anhydrase inhibition assay
In this assay, 20 μL of different concentrations of
MPSE (1-12.5 mg/mL) were added to a well. Along
with the test extract, 20 μL enzyme carbonic
anhydrase solution (0.1 mg/mL) and 140 μL of 20
mM HEPES-tris buffer (pH 7.4) were added to each
well. The 96-well plate was then incubated at 37 °C
for 15 min. After incubation, the absorbance
was measured, and then 20 μL of the substrate
solution 4-nitrophenyl acetate (0.7 mM) prepared
in methanol was added to all wells. The amount
of product formed was monitored by the change
in absorbance for 30 min with intervals of 1 min at
400 nm in a spectrophotometer (Spectra Max, MD,
USA) (26). Acetazolamide served as the positive
(drug) control.
C y t o t o x i c i t y A n a l y s i s U s i n g 3 - ( 4 ,
5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium
bromide (MTT) colorimetric assay
The anticancer activity of the MPSE at different
concentration ranges (1000-10 μg/mL) was measured
in the human neuroblastoma cell line SH-SY5Y.
These cells were grown in Dulbecco’s Modified
Eagle Medium (DMEM) media supplemented with
10% fetal bovine serum and incubated in a 5%
CO2 incubator at 37°C. Upon 75-85% confluency,
cells were detached from the flask and seeded at
5,000 cells/well into 96-well plates. The next day,
after adherence of cells on the plate, these cells
were treated with MPSE at double-fold diluted
concentrations ranging from 100-1 μg/mL. After
24 h, cell viability was checked by adding 20 μL of
MTT (5 mg/mL) dye to each well and the plate was
incubated for 3-4 h. Soon after, formazan crystal
was dissolved by adding 100 μL of 0.1% dimethyl
sulfoxide (DMSO). The plate was shaken for 1 min,
and absorbance was measured at 540 nm using a
spectrophotometer (Tecan, California, USA) (27).
Statistical Analysis
IC 50 value of each fraction was compared with
positive drug control. One-way analysis of variance
(ANOVA) was used in the GraphPad Prism software
version 5.0 and analyzed with a level of significance
(p<0.05).
RESULT
Enzyme inhibitory activity
In the present study, MPSE was tested on different
enzyme systems to evaluate its potential to inhibit
enzymes that cause chronic diseases. At 0.5 mg/
mL, the MPSE revealed anti-enzyme activities
against α-glucosidase, thymidine phosphorylase,
and carbonic anhydrase with inhibitory percentage
values of 89 ± 3, 51.3 ± 0.3, and 80 ± 5 %
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
Fur thermore, the anticancer potential of M.
peregrina seed extract was investigated in the
human neuroblastoma cell line SH-SY5Y.
MATERIALS AND METHODS
Plant Preparation
M. peregrina seeds were submitted and authenticated
by Dr. Maha Kordofani (Resident Botanist) at the
Department of Botany, University of Khartoum. M.
peregrina seeds were kept at room temperature
until they were dried and ground to powder. 100 g
of powder was mixed s with 500 mL of ethanol and
left for 5 days for proper mixing. The macerates were
filtered through a muslin cloth and collected; then,
rotary evaporated to obtain the crude M. peregrina
seed extract (MPSE) (23).
Alpha-Glucosidase inhibition assay
20 μL of MPSE prepared in different concentrations
(0.125-1 mg/mL) were added to a 96-well plate.
Along with the test extract, 20 μL of enzyme solution
(0.2 U/mL) and 135 μL of 0.1 M phosphate buffered
saline (PBS) buffer (pH 6.8) were added to each well.
The plate was then incubated at 37 °C for 15 min.
After incubation, the absorbance was measured
before the substrate addition. The substrate (25 μL)
4-nitrophenyl-α-D-glucopyranoside (PNP-G)
(0.7 mM) was then added, and the change in
absorbance was measured for 30 min at 400 nm in
a spectrophotometer (Spectra Max, MD, USA) (24).
Acarbose served as the positive (drug) control.
Thymidine phosphorylase (TP) inhibition assay
We followed the protocol of Javaid et al. (25). Briefly,
10 μL of MPSE at different concentrations (0.125-1
mg/mL) were incubated with 20 μL of the enzyme
(0.058 unit/well) along with 150 μL of potassium
phosphate buffer (pH 7.0, 50 mM) for 10 min at 30°C.
After 10 min, 20 μL (1.5 mM) substrate (thymidine,
kmax; 265 nm) was added. Soon after, absorbance
was recorded at 290 nm using an ELISA plate reader
and continued for 10 min to monitor the change in
absorbance. 7-Deazaxanthine served as the positive
(drug) control.
Carbonic anhydrase inhibition assay
In this assay, 20 μL of different concentrations of
MPSE (1-12.5 mg/mL) were added to a well. Along
with the test extract, 20 μL enzyme carbonic
anhydrase solution (0.1 mg/mL) and 140 μL of 20
mM HEPES-tris buffer (pH 7.4) were added to each
well. The 96-well plate was then incubated at 37 °C
for 15 min. After incubation, the absorbance
was measured, and then 20 μL of the substrate
solution 4-nitrophenyl acetate (0.7 mM) prepared
in methanol was added to all wells. The amount
of product formed was monitored by the change
in absorbance for 30 min with intervals of 1 min at
400 nm in a spectrophotometer (Spectra Max, MD,
USA) (26). Acetazolamide served as the positive
(drug) control.
C y t o t o x i c i t y A n a l y s i s U s i n g 3 - ( 4 ,
5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium
bromide (MTT) colorimetric assay
The anticancer activity of the MPSE at different
concentration ranges (1000-10 μg/mL) was measured
in the human neuroblastoma cell line SH-SY5Y.
These cells were grown in Dulbecco’s Modified
Eagle Medium (DMEM) media supplemented with
10% fetal bovine serum and incubated in a 5%
CO2 incubator at 37°C. Upon 75-85% confluency,
cells were detached from the flask and seeded at
5,000 cells/well into 96-well plates. The next day,
after adherence of cells on the plate, these cells
were treated with MPSE at double-fold diluted
concentrations ranging from 100-1 μg/mL. After
24 h, cell viability was checked by adding 20 μL of
MTT (5 mg/mL) dye to each well and the plate was
incubated for 3-4 h. Soon after, formazan crystal
was dissolved by adding 100 μL of 0.1% dimethyl
sulfoxide (DMSO). The plate was shaken for 1 min,
and absorbance was measured at 540 nm using a
spectrophotometer (Tecan, California, USA) (27).
Statistical Analysis
IC 50 value of each fraction was compared with
positive drug control. One-way analysis of variance
(ANOVA) was used in the GraphPad Prism software
version 5.0 and analyzed with a level of significance
(p<0.05).
RESULT
Enzyme inhibitory activity
In the present study, MPSE was tested on different
enzyme systems to evaluate its potential to inhibit
enzymes that cause chronic diseases. At 0.5 mg/
mL, the MPSE revealed anti-enzyme activities
against α-glucosidase, thymidine phosphorylase,
and carbonic anhydrase with inhibitory percentage
values of 89 ± 3, 51.3 ± 0.3, and 80 ± 5 %
4Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 31 | Number 01 | Article 349875Shaymaa Fadhel Abbas Albaayit
respectively. The IC 50 of inhibitory activity of MPSE
is shown in Table 1.
Table 1: Enzyme inhibitory activity of the M. peregrina seed
ethanol extract
Enzyme Activity IC50 values
MPSE a standard
α-glucosidase 160.1 ± 5 μg/mL 875.75 ± 2.08 μM
Thymidine phosphorylase 500 ± 6 μg/mL 41.0 ± 1.63 μM
Carbonic anhydrase 180 ± 5 μg/mL 18.2 ± 1.23 μM
a The standard drug Acarbose was used in α-glucosidase assays, 7-Deazaxanthine
in Thymidine phosphorylase, Acetazolamide in Carbonic anhydrase
Anticancer activities
Cytotoxicity was measured by MTT assay. The
result showed that MPSE inhibited the growth of
cancer cells SH-SY5Y, with IC 50 values of 55.1 μg/mL
(Figure 1).
0
50
100
Control 12.5 25 50 100
Cell inhibition %
MPSE μg/ml
Figure 1. The proliferation of neuroblastoma, cell line SH-SY5Y
after treatment with M. peregrina seed extract (MPSE) for 48 h.
DISCUSSION
According to the World Health Organization (WHO)
report, chronic diseases are a silent global epidemic
that accounts for 71% of deaths worldwide (28).
Among these chronic diseases, diabetes is one of
the major risk factors for causing other diseases
related to cardiovascular diseases, respiratory
diseases, cancer, neurodegenerative disorders,
nephropathy, and neuropathy diseases (29). For
the progression of these chronic diseases, enzymes
like carbonic anhydrase, thymidine phosphorylase,
and α-glucosidase play a vital role in the slow
progression of the disease. Therefore, it would be
beneficial to inhibit these enzymes with various
active compounds. In response to these, researchers
have been screening plant extracts against these
enzymes to decrease the disease progression with
fewer side effects.
This present study concluded that MPSE at 0.5
mg/mL showed enzyme inhibitory activity of 89 ±
3, 51.3 ± 0.3, and 80 ± 5 % against α-glucosidase,
thymidine phosphorylase, and carbonic anhydrase,
respectively. Sardabi et al. (30) reported a 50%
inhibition of α-glucosidase by the defatted M.
peregrina seed pressed cake at 30 mg/mL. Ullah et
al. (31) reported the potential of the hydro-alcoholic
extract of M. peregrina to inhibit α-amylase and
α-glucosidase. They attributed these activities to
higher concentrations of total phenols, saponins,
and tannins. Many active compounds have been
reported from M. peregrina, such as polyphenolic
compounds, flavonoids, tocopherols, linoleic acid,
and oleic acid, which may possess enzyme inhibition
activity (32, 33). Koheil et al. (4) reported the
antidiabetic activity of M. peregrina seeds hydro-
alcoholic extract fraction in streptozotocin-induced
diabetic rats. This extract significantly decreased the
blood glucose level at 200 mg/kg of body weight.
The other two fractions (petroleum ether and
chloroform) also decrease the blood glucose level.
In many solid tumors, overexpression of thymidine
phosphorylase induces angiogenesis and prevents
apoptosis of cells leading to the metastatic stage.
Due to its critical function, thymidine phosphorylase
is an ideal target for discovering anti-angiogenic
compounds. In the present study, MPSE showed
inhibition of thymidine phosphorylase enzyme with
an IC 50 of 471 μg/mL.
In solid tumors like neuroblastoma, a hypoxic
condition leads to an acidic microenvironment;
therefore, to survive in this acidic environment, these
continuously dividing cells overexpress carbonic
anhydrase IX and XII isoforms enzymes (35). Over
expressions of these two isoforms maintains a
physiological intracellular pH due to which cells
could survive in acidic extracellular pH. Thus, tumors
progress to the metastatic stage. Due to the critical
role of carbonic anhydrase in the neuroblastoma
progression, it is considered a potential biomarker to
predict the survivability of neuroblastoma patients
(36). Therefore, many researchers have considered
carbonic anhydrase inhibition as the targeted
therapy. In the present study, M. peregrina seed
extract showed promising anti-carbonic anhydrase
inhibition with IC 50 of 271 μg/mL. Besides, MPSE
showed anti-neuroblastoma activity with IC 50 of 55
respectively. The IC 50 of inhibitory activity of MPSE
is shown in Table 1.
Table 1: Enzyme inhibitory activity of the M. peregrina seed
ethanol extract
Enzyme Activity IC50 values
MPSE a standard
α-glucosidase 160.1 ± 5 μg/mL 875.75 ± 2.08 μM
Thymidine phosphorylase 500 ± 6 μg/mL 41.0 ± 1.63 μM
Carbonic anhydrase 180 ± 5 μg/mL 18.2 ± 1.23 μM
a The standard drug Acarbose was used in α-glucosidase assays, 7-Deazaxanthine
in Thymidine phosphorylase, Acetazolamide in Carbonic anhydrase
Anticancer activities
Cytotoxicity was measured by MTT assay. The
result showed that MPSE inhibited the growth of
cancer cells SH-SY5Y, with IC 50 values of 55.1 μg/mL
(Figure 1).
0
50
100
Control 12.5 25 50 100
Cell inhibition %
MPSE μg/ml
Figure 1. The proliferation of neuroblastoma, cell line SH-SY5Y
after treatment with M. peregrina seed extract (MPSE) for 48 h.
DISCUSSION
According to the World Health Organization (WHO)
report, chronic diseases are a silent global epidemic
that accounts for 71% of deaths worldwide (28).
Among these chronic diseases, diabetes is one of
the major risk factors for causing other diseases
related to cardiovascular diseases, respiratory
diseases, cancer, neurodegenerative disorders,
nephropathy, and neuropathy diseases (29). For
the progression of these chronic diseases, enzymes
like carbonic anhydrase, thymidine phosphorylase,
and α-glucosidase play a vital role in the slow
progression of the disease. Therefore, it would be
beneficial to inhibit these enzymes with various
active compounds. In response to these, researchers
have been screening plant extracts against these
enzymes to decrease the disease progression with
fewer side effects.
This present study concluded that MPSE at 0.5
mg/mL showed enzyme inhibitory activity of 89 ±
3, 51.3 ± 0.3, and 80 ± 5 % against α-glucosidase,
thymidine phosphorylase, and carbonic anhydrase,
respectively. Sardabi et al. (30) reported a 50%
inhibition of α-glucosidase by the defatted M.
peregrina seed pressed cake at 30 mg/mL. Ullah et
al. (31) reported the potential of the hydro-alcoholic
extract of M. peregrina to inhibit α-amylase and
α-glucosidase. They attributed these activities to
higher concentrations of total phenols, saponins,
and tannins. Many active compounds have been
reported from M. peregrina, such as polyphenolic
compounds, flavonoids, tocopherols, linoleic acid,
and oleic acid, which may possess enzyme inhibition
activity (32, 33). Koheil et al. (4) reported the
antidiabetic activity of M. peregrina seeds hydro-
alcoholic extract fraction in streptozotocin-induced
diabetic rats. This extract significantly decreased the
blood glucose level at 200 mg/kg of body weight.
The other two fractions (petroleum ether and
chloroform) also decrease the blood glucose level.
In many solid tumors, overexpression of thymidine
phosphorylase induces angiogenesis and prevents
apoptosis of cells leading to the metastatic stage.
Due to its critical function, thymidine phosphorylase
is an ideal target for discovering anti-angiogenic
compounds. In the present study, MPSE showed
inhibition of thymidine phosphorylase enzyme with
an IC 50 of 471 μg/mL.
In solid tumors like neuroblastoma, a hypoxic
condition leads to an acidic microenvironment;
therefore, to survive in this acidic environment, these
continuously dividing cells overexpress carbonic
anhydrase IX and XII isoforms enzymes (35). Over
expressions of these two isoforms maintains a
physiological intracellular pH due to which cells
could survive in acidic extracellular pH. Thus, tumors
progress to the metastatic stage. Due to the critical
role of carbonic anhydrase in the neuroblastoma
progression, it is considered a potential biomarker to
predict the survivability of neuroblastoma patients
(36). Therefore, many researchers have considered
carbonic anhydrase inhibition as the targeted
therapy. In the present study, M. peregrina seed
extract showed promising anti-carbonic anhydrase
inhibition with IC 50 of 271 μg/mL. Besides, MPSE
showed anti-neuroblastoma activity with IC 50 of 55
5Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 31 | Number 01 | Article 349875
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
μg/mL. It was reported that carbonic anhydrase
inhibitor increases the anti-neuroblastoma effect
of cisplatin through a synergistic effect (37);
therefore, MPSE combined with cisplatin may show
a synergistic effect. Thus, this study paved the way
to study the potential of MPSE and its synergistic
effect with cisplatin in treating neuroblastoma.
M. peregrina seed extract inhibited three different
enzymes: α-glucosidase, carbonic anhydrase, and
thymidine phosphorylase. M. peregrina was also
reported to possess enzyme inhibition activity
against urease (23), dipeptidyl peptidase IV (31),
and angiotensin-converting enzyme (38). Thus, this
plant could be used in treating different chronic
diseases caused due to the overexpression of these
enzymes (39).
M. peregrina seeds have been reported for their
cytotoxic effect on cell lines HeLa, MCF-7, CaCO2,
and HepG2 (40). The anti-cancer activity showed
by MPSE against these cancer cell lines, and SH-
SY5Y might be due to the presence of phenolic/
flavonoid compounds (41-44) like catechin, catechol,
resveratrol, coumarin, naringin, rutin, quercetin,
kaempferol, hispertin, and apigenin, which induce
apoptosis by different pathways like activation of the
mitochondrial pathway, and caspase-3 dependent
apoptotic pathway, downregulation of anti-apoptotic
genes (Bcl-2, Bcl-xL), and upregulation of caspase3
with the release of cytochrome c., premature aging,
cell cycle arrest, and enhance the immune system
to engulf cancer cells (45,48).
CONCLUSION
M. peregrina seed extract showed good enzyme
inhibition against α-glucosidase and carbonic
anhydrase with IC 50 value below 200 μg/mL. M.
peregrina also revealed anti-neuroblastoma activity,
which supported its anticancer property against
other cancer cell lines. The results showed carbonic
anhydrase inhibitor along with anti-neuroblastoma
effect of the extract. Therefore, M. peregrina could
be combined with available anticancer drugs to
enhance their anticancer effects.
ACKNOWLEDGEMENT
The Corresponding author highly appreciate and
thankful to Prof. Dr. M. Iqbal Choudhary (Director,
ICCBS) for providing NAM-ICCBS fellowship in
ICCBS, University of Karachi, Pakistan. Author also
thankful to Dr. Rukesh Maharjan (ICCBS) for his help
and guidance.
REFERENCES
[1] Albaayit SFA. Enzyme inhibitory properties of Zerumbone. Pak
J Agri Sci. 2021;58:1207-1209. DOI: https://doi.org/10.21162/
PAKJAS/21.9759
[2] Rauf A, Jehan N. Natural products a as potential enzyme
inhibitors from medicinal plants. Enzyme Inhib Activators.
2017;165:177. DOI: https://doi.org/10.5772/67376
[3] Anand U, Jacobo-Herrera N, Altemimi A, Lakhssassi N. A
comprehensive review on medicinal plants as antimicrobial
therapeutics: potential avenues of biocompatible drug discovery.
Metabolites. 2019;9(11):258. DOI: https://doi.org/10.3390/
metabo9110258
[4] Albaayit SFA, Rasedee A, Noor MHM. Zerumbone-Loaded
Nanostructured Lipid Carrier Gel Enhances Wound Healing
in Diabetic Rats. BioMed Res Int. 2022. DOI: https://doi.
org/10.1155/2022/1129297
[5] Humadi S, Hassan SM, Ahjel SW. A Cross-Sectional Survey
of Iraqi Herbalist Practicing in the Middle Euphrates Area
with Recognition of their Knowledge, Practice and Attitude
(Conference Paper). Iraq Pharm Sci. 2022;31:178-187. DOI: https://
doi.org/10.31351/vol31issSuppl.pp178-187
[6] A lb aayit SFA . Evaluatio n of anti - met hicillin resis t ant
Staphylococcus aureus property of Clausena excavata leaves
by using atomic force microscopy and flowcytometry techniques.
Pak J Agri Sci. 2021;58:315-320. DOI: https://doi.org/10.21162/
PAKJAS/21.921
[7] Shaito A, Thuan DT, Phu HT, Nguyen TH, Hasan H, Halabi S,
Abdelhady S, Nasrallah GK, Eid AH, Pintus G. Herbal medicine
for cardiovascular diseases: efficacy, mechanisms, and safety.
Front pharmacol. 2020;11:422. DOI: https://doi.org/10.3389/
fphar.2020.00422
[8] Albaayit SFA, Maharjan R, Abdullah R, Noor MHM. Evaluation
of antimethicillin-resistant Staphylococcus aureus property of
zerumbone. J Appl Biomed. 2022;20:22-36.
[9] Senthilkumar A, Karuvantevida N, Rastrelli L, Kurup SS, Cheruth AJ.
Traditional uses, pharmacological efficacy, and phytochemistry
of Moringa peregrina (Forssk.) Fiori. A Review. Front Pharmacol.
2018;9:465. DOI: https://doi.org/10.3389/fphar.2018.00465
[10] Elbatran SA, Abdel-Salam OM, Abdelshfeek KA, Nazif NM,
Ismail SI, Hammouda FM. Phytochemical and pharmacological
investigations on Moringa peregrina (Forssk) Fiori. Nat Prod Sci.
2005;11(4):199-206. DOI: https://doi.org/10.32725/jab.2022.002
[11] El - H a d d a d A E, Ko h e i l M A , El - K h a l i k SM, O s m a n S .
Antihyperglycemic activity and nitrile glycosides of Moringa
peregrina (Forssk.) seeds. In Four th Euro-Mediterranean
Conference of Natural Products and Drug Discovery: Back to
Mother Nature (Cairo/Sharm El-Sheikh: BioNat-IV). 2002. DOI:
https://doi.org/10.13140/RG.2.2.25894.93763
[12] Albaayit SFA, Al-Khafaji ASK, Alnaimy HS. In vitro macrophage
nitric oxide and interleukin-1 beta suppression by Moringa
peregrina seed. Turk J Pharm Sci. 2019;16(3):362-365. DOI:
https://doi.org/10.4274/tjps.galenos.2018.52244
[13] Al-Owaisi M, Al-Hadiwi N, Khan SA. GC-MS analysis, determination
of total phenolics, flavonoid content and free radical scavenging
activities of various crude extracts of Moringa peregrina (Forssk.)
Fiori leaves. Asian Pacific J Trop Biomed. 2014;4(12):964-970. DOI:
https://doi.org/10.12980/APJTB.4.201414B295
[14] Asghari G, Palizban A, Bakhshaei B. Quantitative analysis
of the nutritional components in leaves and seeds of the
Persian Moringa peregrina (Forssk.) Fiori. Pharmacognosy Res.
2015;7(3):242. DOI: https://doi.org/10.4103/0974-8490.157968
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
μg/mL. It was reported that carbonic anhydrase
inhibitor increases the anti-neuroblastoma effect
of cisplatin through a synergistic effect (37);
therefore, MPSE combined with cisplatin may show
a synergistic effect. Thus, this study paved the way
to study the potential of MPSE and its synergistic
effect with cisplatin in treating neuroblastoma.
M. peregrina seed extract inhibited three different
enzymes: α-glucosidase, carbonic anhydrase, and
thymidine phosphorylase. M. peregrina was also
reported to possess enzyme inhibition activity
against urease (23), dipeptidyl peptidase IV (31),
and angiotensin-converting enzyme (38). Thus, this
plant could be used in treating different chronic
diseases caused due to the overexpression of these
enzymes (39).
M. peregrina seeds have been reported for their
cytotoxic effect on cell lines HeLa, MCF-7, CaCO2,
and HepG2 (40). The anti-cancer activity showed
by MPSE against these cancer cell lines, and SH-
SY5Y might be due to the presence of phenolic/
flavonoid compounds (41-44) like catechin, catechol,
resveratrol, coumarin, naringin, rutin, quercetin,
kaempferol, hispertin, and apigenin, which induce
apoptosis by different pathways like activation of the
mitochondrial pathway, and caspase-3 dependent
apoptotic pathway, downregulation of anti-apoptotic
genes (Bcl-2, Bcl-xL), and upregulation of caspase3
with the release of cytochrome c., premature aging,
cell cycle arrest, and enhance the immune system
to engulf cancer cells (45,48).
CONCLUSION
M. peregrina seed extract showed good enzyme
inhibition against α-glucosidase and carbonic
anhydrase with IC 50 value below 200 μg/mL. M.
peregrina also revealed anti-neuroblastoma activity,
which supported its anticancer property against
other cancer cell lines. The results showed carbonic
anhydrase inhibitor along with anti-neuroblastoma
effect of the extract. Therefore, M. peregrina could
be combined with available anticancer drugs to
enhance their anticancer effects.
ACKNOWLEDGEMENT
The Corresponding author highly appreciate and
thankful to Prof. Dr. M. Iqbal Choudhary (Director,
ICCBS) for providing NAM-ICCBS fellowship in
ICCBS, University of Karachi, Pakistan. Author also
thankful to Dr. Rukesh Maharjan (ICCBS) for his help
and guidance.
REFERENCES
[1] Albaayit SFA. Enzyme inhibitory properties of Zerumbone. Pak
J Agri Sci. 2021;58:1207-1209. DOI: https://doi.org/10.21162/
PAKJAS/21.9759
[2] Rauf A, Jehan N. Natural products a as potential enzyme
inhibitors from medicinal plants. Enzyme Inhib Activators.
2017;165:177. DOI: https://doi.org/10.5772/67376
[3] Anand U, Jacobo-Herrera N, Altemimi A, Lakhssassi N. A
comprehensive review on medicinal plants as antimicrobial
therapeutics: potential avenues of biocompatible drug discovery.
Metabolites. 2019;9(11):258. DOI: https://doi.org/10.3390/
metabo9110258
[4] Albaayit SFA, Rasedee A, Noor MHM. Zerumbone-Loaded
Nanostructured Lipid Carrier Gel Enhances Wound Healing
in Diabetic Rats. BioMed Res Int. 2022. DOI: https://doi.
org/10.1155/2022/1129297
[5] Humadi S, Hassan SM, Ahjel SW. A Cross-Sectional Survey
of Iraqi Herbalist Practicing in the Middle Euphrates Area
with Recognition of their Knowledge, Practice and Attitude
(Conference Paper). Iraq Pharm Sci. 2022;31:178-187. DOI: https://
doi.org/10.31351/vol31issSuppl.pp178-187
[6] A lb aayit SFA . Evaluatio n of anti - met hicillin resis t ant
Staphylococcus aureus property of Clausena excavata leaves
by using atomic force microscopy and flowcytometry techniques.
Pak J Agri Sci. 2021;58:315-320. DOI: https://doi.org/10.21162/
PAKJAS/21.921
[7] Shaito A, Thuan DT, Phu HT, Nguyen TH, Hasan H, Halabi S,
Abdelhady S, Nasrallah GK, Eid AH, Pintus G. Herbal medicine
for cardiovascular diseases: efficacy, mechanisms, and safety.
Front pharmacol. 2020;11:422. DOI: https://doi.org/10.3389/
fphar.2020.00422
[8] Albaayit SFA, Maharjan R, Abdullah R, Noor MHM. Evaluation
of antimethicillin-resistant Staphylococcus aureus property of
zerumbone. J Appl Biomed. 2022;20:22-36.
[9] Senthilkumar A, Karuvantevida N, Rastrelli L, Kurup SS, Cheruth AJ.
Traditional uses, pharmacological efficacy, and phytochemistry
of Moringa peregrina (Forssk.) Fiori. A Review. Front Pharmacol.
2018;9:465. DOI: https://doi.org/10.3389/fphar.2018.00465
[10] Elbatran SA, Abdel-Salam OM, Abdelshfeek KA, Nazif NM,
Ismail SI, Hammouda FM. Phytochemical and pharmacological
investigations on Moringa peregrina (Forssk) Fiori. Nat Prod Sci.
2005;11(4):199-206. DOI: https://doi.org/10.32725/jab.2022.002
[11] El - H a d d a d A E, Ko h e i l M A , El - K h a l i k SM, O s m a n S .
Antihyperglycemic activity and nitrile glycosides of Moringa
peregrina (Forssk.) seeds. In Four th Euro-Mediterranean
Conference of Natural Products and Drug Discovery: Back to
Mother Nature (Cairo/Sharm El-Sheikh: BioNat-IV). 2002. DOI:
https://doi.org/10.13140/RG.2.2.25894.93763
[12] Albaayit SFA, Al-Khafaji ASK, Alnaimy HS. In vitro macrophage
nitric oxide and interleukin-1 beta suppression by Moringa
peregrina seed. Turk J Pharm Sci. 2019;16(3):362-365. DOI:
https://doi.org/10.4274/tjps.galenos.2018.52244
[13] Al-Owaisi M, Al-Hadiwi N, Khan SA. GC-MS analysis, determination
of total phenolics, flavonoid content and free radical scavenging
activities of various crude extracts of Moringa peregrina (Forssk.)
Fiori leaves. Asian Pacific J Trop Biomed. 2014;4(12):964-970. DOI:
https://doi.org/10.12980/APJTB.4.201414B295
[14] Asghari G, Palizban A, Bakhshaei B. Quantitative analysis
of the nutritional components in leaves and seeds of the
Persian Moringa peregrina (Forssk.) Fiori. Pharmacognosy Res.
2015;7(3):242. DOI: https://doi.org/10.4103/0974-8490.157968
6Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 31 | Number 01 | Article 349875Shaymaa Fadhel Abbas Albaayit
[15] Afsharypuor S, Asghari G, Mohagheghzadeh A, Dehshahri S.
Volatile constituents of the seed kernel and leaf of Moringa
peregrina (Forssk.) Fiori, Agricolt. Cultivated in Chabahar (Iran).
Iran J Pharm Sci. 2010;6(2):141-144. DOI: https://www.ijps.ir/
article_2138_1bb26f237636da67948db0051e24476a.pdf
[16] Qaisar MN, Chaudhary BA, Sajid MU, Hussain N. Evaluation
of α-glucosidase inhibitory activity of dichloromethane and
methanol extracts of Croton bonplandianum Baill. Trop J Pharm
Res. 2014;13(11):1833-1836. DOI: https://doi.org/ 10.4314/tjpr.
v13i11.9
[17] Elamin YY, Rafee S, Osman N, Gately K. Thymidine phosphorylase
in cancer; enemy or friend?. Cancer Microenviron. 2016; 9(1):33-
43. DOI: https://doi.org/10.1007/s12307-015-0173-y
[18] Gaaib JN, Jumaa MG. Expression of Carbonic Anhydrase IX
(CA9) in Human Tumor Tissue of Patients with Ovarian Cancer.
Iraqi journal of biotechnology. 2017;16(2):81-88. DOI: https://jige.
uobaghdad.edu.iq/index.php/IJB/article/view/93/56
[19] Albaayit SFA, Mohammed MK. Antiangiogenic activity and ROS-
mediated lung cancer cell line injury of zerumbone. J Fac Pharm
Ankara. 2023;47(3). DOI: https://doi.org/10.33483/jfpau.1112778
[20] Albaayit SFA, Maharjan R, Abdullah R, Noor MHM. Anti-
Enterococcus Faecalis, Cytotoxicity, Phytotoxicity, and Anticancer
Studies on Clausena excavata Burum. f.(Rutaceae) Leaves.
BioMed Res Int. 2021. DOI: https://doi.org/10.1155/2021/3123476
[21] Albaayit SFA, Khan M, Abdullah R. Zerumbone induces
growth inhibition of Burkitt’s lymphoma cell line via apoptosis.
Nat Volatiles Essent Oils. 2021;8(3):56-63. DOI: https://doi.
org/10.37929/nveo.927770
[22] Albaayit SFA, Maharjan R, Khan M. Evaluation of Hemolysis
Activity of Zerumbone on RBCs and Brine Shrimp Toxicity.
Baghdad Sci J. 2021;18:65-69. DOI: https://doi.org/10.21123/
bsj.2021.18.1.0065
[23] Albaayit SFA, Ozaslan M. Cytotoxic and urease inhibition
potential of Moringa peregrina seed ethanolic extract. Int J
Pharmacol. 2019;15(1):151-155. DOI: https://doi.org/10.3923/
ijp.2019.151.155
[24] Maher S, Choudhary MI, Saleem F, Rasheed S, Waheed I, Halim
SA, Azeem M, Abdullah IB, Froeyen M, Mirza MU, Ahmad S.
Isolation of antidiabetic withanolides from Withania coagulans
Dunal and their in vitro and in silico validation. Biology.
2020;9(8):197. DOI: https://doi.org/10.3390/biology9080197
[25] Javaid S, Saad SM, Perveen S, Khan KM, Choudhary MI.
2-Ar ylquinazolin-4 (3H)-ones: A novel class of thymidine
phosphorylase inhibitors. Bioorg Chem. 2015;63:142-151. DOI:
https://doi.org/10.1016/j.bioorg.2015.10.006
[26] Avula SK, Rehman NU, Khan M, Halim SA, Khan A, Rafiq K,
Csuk R, Das B, Al-Harrasi A. New synthetic 1H-1, 2, 3-triazole
derivatives of 3-O-acetyl-β-boswellic acid and 3-O-acetyl-11-
keto-β-boswellic acid from Boswellia sacra inhibit carbonic
anhydrase II in vitro. Med Chem Res. 2021;30(6):1185-1198. DOI:
https://doi.org/10.1007/s00044-021-02723-8
[27] Hussain SS, Rafi K, Faizi S, Razzak ZA, Simjee SU. A novel, semi-
synthetic diterpenoid 16 (R and S)-phenylamino-cleroda-3, 13
(14), Z-dien-15, 16 olide (PGEA-AN) inhibits the growth and cell
survival of human neuroblastoma cell line SH-SY5Y by modulating
P53 pathway. Mol Cell Biochem. 2018;449(1):105-115. DOI:
https://doi.org/10.1007/s11010-018-3347-3
[28] WHO Noncommunicable Diseases. 2021. [(accessed on 31
December 2021)]. Available online: https://www.who.int/news-
room/fact-sheets/detail/noncommunicable-diseases
[29] Cole JB, Florez JC. Florez. Genetics of diabetes mellitus and
diabetes complications. Nat Rev Nephrol. 2010;16(7):377-390.
DOI: https://doi.org/10.1038/s41581-020-0278-5
[30] Sardabi F, Azizi MH, Gavlighi HA, Rashidinejad A. Potential
benefits of Moringa peregrina defatted seed: Effect of processing
on nutritional and anti-nutritional properties, antioxidant
capacity, in vitro digestibility of protein and starch, and inhibition
of α-glucosidase and α-amylase enzymes. Food Chem Adv.
2022:100034. DOI: https://doi.org/10.1016/j.focha.2022.100034
[31] Ullah MF, Bhat SH, Abuduhier FM. Antidiabetic Potential
of Hydro‐Alcoholic Extract of M oringa Peregrina Leaves:
Implication as Functional Food for Prophylactic Intervention in
Prediabetic Stage. J. Food Biochem. 2015;39(4):360-367. DOI:
https://doi.org/10.1111/jfbc.12140
[32] ŞöhretoŞlu D, Sari S. Flavonoids as alpha-glucosidase inhibitors:
Mechanistic approaches merged with enzyme kinetics and
molecular modelling. Phytochem Rev. 2010;19(5):1081-1092.
https://doi.org/10.1007/s11101-019-09610-6
[33] Su CH, Hsu CH, Ng LT. Inhibitory potential of fatty acids on key
enzymes related to type 2 diabetes. Biofactors. 2013;39(4):415-
421. DOI: https://doi.org/10.1002/biof.1082
[34] Koheil MA, Hussein MA, Othman SM, El-Haddad A. In-vivo
antioxidant activity of Moringa peregrina against STZ–induced
oxidative stress in type 2 diabetic rats. Mol Clin Pharmacol.
2013;4:65-75.
[35] Cimmino F, Pezone L, Avitabile M, Persano L, Vitale M, Sassi M,
Bresolin S, Serafin V, Zambrano N, Scaloni A, Basso G. Proteomic
alterations in response to hypoxia inducible factor 2α in normoxic
neuroblastoma cells. J. Proteome Res. 2016;15(10):3643-3655.
DOI: https://doi.org/10.1021/acs.jproteome.6b00457
[36] Ameis HM, Drenckhan A, Freytag M, Izbicki JR, Supuran CT,
Reinshagen K, Holland-Cunz S, Gros SJ. Carbonic anhydrase IX
correlates with survival and is a potential therapeutic target for
neuroblastoma. J Enzyme Inhib Med Chem. 2016;31(3):404-409.
DOI: https://doi.org/10.3109/14756366.2015.1029471
[37] Garbati P, Barbieri R, Cangelosi D, Zanon C, Costa D, Eva
A, Thellung S, Calderoni M, Baldini F, Tonini GP, Modesto
P. MCM2 and carbonic anhydrase 9 are novel potential
target s for neuroblastoma pharmacological treatment.
Biomedicines. 2020;8(11):471. DOI: https://doi.org/10.3390/
biomedicines8110471
[38] Gonçalves S, Romano A. Inhibition of angiotensin converting
enzyme by Rhazya stricta, Moringa peregrina and Achillea
fragrantissima, used in traditional system of medicine in Arabian
Peninsula: Implication in the management of hypertension.
J Med Plant Res. 2016;10(8):93-99. DOI: https://doi.org/10.5897/
JMPR2015.6043
[39] Gonçalves S, Romano A. Inhibitory properties of phenolic
compounds against enzymes linked with human diseases.
Phenolic compounds-biological activity. London: Intech Open.
2017;99-118. DOI: https://doi.org/10.5772/66844
[40] Albaayit SFA. In vitro evaluation of anticancer activity of
Moringa peregrina seeds on breast cancer cells. Eurasia J Math
Sci. 2020;11:163-166. DOI: http://www.epstem.net/en/pub/
issue/58065/838264
[41] Hazafa A, Rehman KU, Jahan N, Jabeen Z. The role of polyphenol
(flavonoids) compounds in the treatment of cancer cells. Nutr
Cancer. 2020;72(3):386-397. DOI: https://doi.org/10.1080/0163
5581.2019.1637006
[42] Abd Rani NZ, Husain K, Kumolosasi E. Moringa genus: a review of
phytochemistry and pharmacology. Front Pharmacol. 2018;9:108.
DOI: https://doi.org/10.3389/fphar.2018.00108
[43] Abo El-Fadl S, Osman A, Al-Zohairy AM, Dahab AA, Abo El Kheir
ZA. Assessment of total phenolic, flavonoid content, antioxidant
potential and hplc profile of three moringa species leaf extracts.
sci J flowers Ornam Plants. 2020;7(1):53-70. DOI: https://doi.
org/10.21608/sjfop.2020.91397
[15] Afsharypuor S, Asghari G, Mohagheghzadeh A, Dehshahri S.
Volatile constituents of the seed kernel and leaf of Moringa
peregrina (Forssk.) Fiori, Agricolt. Cultivated in Chabahar (Iran).
Iran J Pharm Sci. 2010;6(2):141-144. DOI: https://www.ijps.ir/
article_2138_1bb26f237636da67948db0051e24476a.pdf
[16] Qaisar MN, Chaudhary BA, Sajid MU, Hussain N. Evaluation
of α-glucosidase inhibitory activity of dichloromethane and
methanol extracts of Croton bonplandianum Baill. Trop J Pharm
Res. 2014;13(11):1833-1836. DOI: https://doi.org/ 10.4314/tjpr.
v13i11.9
[17] Elamin YY, Rafee S, Osman N, Gately K. Thymidine phosphorylase
in cancer; enemy or friend?. Cancer Microenviron. 2016; 9(1):33-
43. DOI: https://doi.org/10.1007/s12307-015-0173-y
[18] Gaaib JN, Jumaa MG. Expression of Carbonic Anhydrase IX
(CA9) in Human Tumor Tissue of Patients with Ovarian Cancer.
Iraqi journal of biotechnology. 2017;16(2):81-88. DOI: https://jige.
uobaghdad.edu.iq/index.php/IJB/article/view/93/56
[19] Albaayit SFA, Mohammed MK. Antiangiogenic activity and ROS-
mediated lung cancer cell line injury of zerumbone. J Fac Pharm
Ankara. 2023;47(3). DOI: https://doi.org/10.33483/jfpau.1112778
[20] Albaayit SFA, Maharjan R, Abdullah R, Noor MHM. Anti-
Enterococcus Faecalis, Cytotoxicity, Phytotoxicity, and Anticancer
Studies on Clausena excavata Burum. f.(Rutaceae) Leaves.
BioMed Res Int. 2021. DOI: https://doi.org/10.1155/2021/3123476
[21] Albaayit SFA, Khan M, Abdullah R. Zerumbone induces
growth inhibition of Burkitt’s lymphoma cell line via apoptosis.
Nat Volatiles Essent Oils. 2021;8(3):56-63. DOI: https://doi.
org/10.37929/nveo.927770
[22] Albaayit SFA, Maharjan R, Khan M. Evaluation of Hemolysis
Activity of Zerumbone on RBCs and Brine Shrimp Toxicity.
Baghdad Sci J. 2021;18:65-69. DOI: https://doi.org/10.21123/
bsj.2021.18.1.0065
[23] Albaayit SFA, Ozaslan M. Cytotoxic and urease inhibition
potential of Moringa peregrina seed ethanolic extract. Int J
Pharmacol. 2019;15(1):151-155. DOI: https://doi.org/10.3923/
ijp.2019.151.155
[24] Maher S, Choudhary MI, Saleem F, Rasheed S, Waheed I, Halim
SA, Azeem M, Abdullah IB, Froeyen M, Mirza MU, Ahmad S.
Isolation of antidiabetic withanolides from Withania coagulans
Dunal and their in vitro and in silico validation. Biology.
2020;9(8):197. DOI: https://doi.org/10.3390/biology9080197
[25] Javaid S, Saad SM, Perveen S, Khan KM, Choudhary MI.
2-Ar ylquinazolin-4 (3H)-ones: A novel class of thymidine
phosphorylase inhibitors. Bioorg Chem. 2015;63:142-151. DOI:
https://doi.org/10.1016/j.bioorg.2015.10.006
[26] Avula SK, Rehman NU, Khan M, Halim SA, Khan A, Rafiq K,
Csuk R, Das B, Al-Harrasi A. New synthetic 1H-1, 2, 3-triazole
derivatives of 3-O-acetyl-β-boswellic acid and 3-O-acetyl-11-
keto-β-boswellic acid from Boswellia sacra inhibit carbonic
anhydrase II in vitro. Med Chem Res. 2021;30(6):1185-1198. DOI:
https://doi.org/10.1007/s00044-021-02723-8
[27] Hussain SS, Rafi K, Faizi S, Razzak ZA, Simjee SU. A novel, semi-
synthetic diterpenoid 16 (R and S)-phenylamino-cleroda-3, 13
(14), Z-dien-15, 16 olide (PGEA-AN) inhibits the growth and cell
survival of human neuroblastoma cell line SH-SY5Y by modulating
P53 pathway. Mol Cell Biochem. 2018;449(1):105-115. DOI:
https://doi.org/10.1007/s11010-018-3347-3
[28] WHO Noncommunicable Diseases. 2021. [(accessed on 31
December 2021)]. Available online: https://www.who.int/news-
room/fact-sheets/detail/noncommunicable-diseases
[29] Cole JB, Florez JC. Florez. Genetics of diabetes mellitus and
diabetes complications. Nat Rev Nephrol. 2010;16(7):377-390.
DOI: https://doi.org/10.1038/s41581-020-0278-5
[30] Sardabi F, Azizi MH, Gavlighi HA, Rashidinejad A. Potential
benefits of Moringa peregrina defatted seed: Effect of processing
on nutritional and anti-nutritional properties, antioxidant
capacity, in vitro digestibility of protein and starch, and inhibition
of α-glucosidase and α-amylase enzymes. Food Chem Adv.
2022:100034. DOI: https://doi.org/10.1016/j.focha.2022.100034
[31] Ullah MF, Bhat SH, Abuduhier FM. Antidiabetic Potential
of Hydro‐Alcoholic Extract of M oringa Peregrina Leaves:
Implication as Functional Food for Prophylactic Intervention in
Prediabetic Stage. J. Food Biochem. 2015;39(4):360-367. DOI:
https://doi.org/10.1111/jfbc.12140
[32] ŞöhretoŞlu D, Sari S. Flavonoids as alpha-glucosidase inhibitors:
Mechanistic approaches merged with enzyme kinetics and
molecular modelling. Phytochem Rev. 2010;19(5):1081-1092.
https://doi.org/10.1007/s11101-019-09610-6
[33] Su CH, Hsu CH, Ng LT. Inhibitory potential of fatty acids on key
enzymes related to type 2 diabetes. Biofactors. 2013;39(4):415-
421. DOI: https://doi.org/10.1002/biof.1082
[34] Koheil MA, Hussein MA, Othman SM, El-Haddad A. In-vivo
antioxidant activity of Moringa peregrina against STZ–induced
oxidative stress in type 2 diabetic rats. Mol Clin Pharmacol.
2013;4:65-75.
[35] Cimmino F, Pezone L, Avitabile M, Persano L, Vitale M, Sassi M,
Bresolin S, Serafin V, Zambrano N, Scaloni A, Basso G. Proteomic
alterations in response to hypoxia inducible factor 2α in normoxic
neuroblastoma cells. J. Proteome Res. 2016;15(10):3643-3655.
DOI: https://doi.org/10.1021/acs.jproteome.6b00457
[36] Ameis HM, Drenckhan A, Freytag M, Izbicki JR, Supuran CT,
Reinshagen K, Holland-Cunz S, Gros SJ. Carbonic anhydrase IX
correlates with survival and is a potential therapeutic target for
neuroblastoma. J Enzyme Inhib Med Chem. 2016;31(3):404-409.
DOI: https://doi.org/10.3109/14756366.2015.1029471
[37] Garbati P, Barbieri R, Cangelosi D, Zanon C, Costa D, Eva
A, Thellung S, Calderoni M, Baldini F, Tonini GP, Modesto
P. MCM2 and carbonic anhydrase 9 are novel potential
target s for neuroblastoma pharmacological treatment.
Biomedicines. 2020;8(11):471. DOI: https://doi.org/10.3390/
biomedicines8110471
[38] Gonçalves S, Romano A. Inhibition of angiotensin converting
enzyme by Rhazya stricta, Moringa peregrina and Achillea
fragrantissima, used in traditional system of medicine in Arabian
Peninsula: Implication in the management of hypertension.
J Med Plant Res. 2016;10(8):93-99. DOI: https://doi.org/10.5897/
JMPR2015.6043
[39] Gonçalves S, Romano A. Inhibitory properties of phenolic
compounds against enzymes linked with human diseases.
Phenolic compounds-biological activity. London: Intech Open.
2017;99-118. DOI: https://doi.org/10.5772/66844
[40] Albaayit SFA. In vitro evaluation of anticancer activity of
Moringa peregrina seeds on breast cancer cells. Eurasia J Math
Sci. 2020;11:163-166. DOI: http://www.epstem.net/en/pub/
issue/58065/838264
[41] Hazafa A, Rehman KU, Jahan N, Jabeen Z. The role of polyphenol
(flavonoids) compounds in the treatment of cancer cells. Nutr
Cancer. 2020;72(3):386-397. DOI: https://doi.org/10.1080/0163
5581.2019.1637006
[42] Abd Rani NZ, Husain K, Kumolosasi E. Moringa genus: a review of
phytochemistry and pharmacology. Front Pharmacol. 2018;9:108.
DOI: https://doi.org/10.3389/fphar.2018.00108
[43] Abo El-Fadl S, Osman A, Al-Zohairy AM, Dahab AA, Abo El Kheir
ZA. Assessment of total phenolic, flavonoid content, antioxidant
potential and hplc profile of three moringa species leaf extracts.
sci J flowers Ornam Plants. 2020;7(1):53-70. DOI: https://doi.
org/10.21608/sjfop.2020.91397
7Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 31 | Number 01 | Article 349875
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
[44] Albaayit SFA, Rasedee A, Abdullah N, Abba Y. Methanolic
extract of Clausena excavata promotes wound healing via
antiinflammatory and anti-apoptotic activities. Asian Pac J Trop
Biomed. 2020;10(5):232-238. DOI: https://doi.org/10.4103/2221-
1691.281467
[45] Saidu NE, Valente S, Bana E, Kirsch G, Bagrel D, Montenarh M.
Coumarin polysulfides inhibit cell growth and induce apoptosis
in HCT116 colon cancer cells. Bioorg Med Chem. 2012;20(4):
1584-1593. DOI: https://doi.org/10.1016/j.bmc.2011.12.032
[46] Albaayit SFA, Khan M, Rasedee A, Noor MHM. Ethyl acetate extract
of Clausena excavata induces growth inhibition of non-small-lung
cancer, NCI-H460, cell line via apoptosis. J Appl Biomed. 2021;19:
40-47. DOI: https://doi.org/10.32725/jab.2021.007
[47] Assim MM, Saheb EJ. The Association of Severe Toxoplasmosis
and Some Cytokine Levels in Breast Cancer Patients. Iraqi J Sci.
2018;59:1189-1194. https://doi.org/0.24996/ijs.2018.59.3A.6
[48] Lafta FM, AL-Jumaily R MK, Rasoul LM. Global DNA Methylation
Levels in Epstein-Barr-Virus-Positive Iraqi Patients with Acute
Lymphoblastic Leukaemia. Iraqi J Sci. 2023;64(3):1109–1118.
Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina
[44] Albaayit SFA, Rasedee A, Abdullah N, Abba Y. Methanolic
extract of Clausena excavata promotes wound healing via
antiinflammatory and anti-apoptotic activities. Asian Pac J Trop
Biomed. 2020;10(5):232-238. DOI: https://doi.org/10.4103/2221-
1691.281467
[45] Saidu NE, Valente S, Bana E, Kirsch G, Bagrel D, Montenarh M.
Coumarin polysulfides inhibit cell growth and induce apoptosis
in HCT116 colon cancer cells. Bioorg Med Chem. 2012;20(4):
1584-1593. DOI: https://doi.org/10.1016/j.bmc.2011.12.032
[46] Albaayit SFA, Khan M, Rasedee A, Noor MHM. Ethyl acetate extract
of Clausena excavata induces growth inhibition of non-small-lung
cancer, NCI-H460, cell line via apoptosis. J Appl Biomed. 2021;19:
40-47. DOI: https://doi.org/10.32725/jab.2021.007
[47] Assim MM, Saheb EJ. The Association of Severe Toxoplasmosis
and Some Cytokine Levels in Breast Cancer Patients. Iraqi J Sci.
2018;59:1189-1194. https://doi.org/0.24996/ijs.2018.59.3A.6
[48] Lafta FM, AL-Jumaily R MK, Rasoul LM. Global DNA Methylation
Levels in Epstein-Barr-Virus-Positive Iraqi Patients with Acute
Lymphoblastic Leukaemia. Iraqi J Sci. 2023;64(3):1109–1118.