This study aimed to compare the antiangiogenic activity of guyabano compared with bevacizumab and paclitaxel, which are among the most common agents known to target angiogenesis in cancer patients.

1. A. muricata leaf extract, paclitaxel,
a n d b e v a c i z u m a b e x h i b i t e d
antiangiogenic activities of different
levels on CAM assay. After comparing
to the negative control 0.2% DMSO, it
was noted that A. muricata leaf extract
exhibited antiangiogenic effect, but was
the least antiangiogenic among the
other treatments. On the other hand,
bevacizumab exhibited the most
significant antiangiogenic activity.
These findings suggest that despite
the capability of A. muricata to exhibit
a n t i a n g i o g e n e s i s , i t i s s t i l l
recommended to use standard agents
like bevacizumab or paclitaxel in
treating cancer, instead of relying on the
minimal antiangiogenic activity of A.
muricata found in food supplements.
Angiogenesis is one of the hallmarks
of cancer and is a target of new drugs.
For the past years, Annona muricata
(guyabano) supplements (Fig.1) have
dominated the Philippine market after a
paper funded by the Department of
Science and Technology revealed that it
has anti-angiogenic activity on chick
chorioallantoic membrane (CAM)
assays. However, to date, there are no
human cancer studies to support this
claim. This study aimed to compare the
antiangiogenic activity of guyabano
compared with bevacizumab and
paclitaxel, which are among the most
common agents known to target
angiogenesis in cancer patients.
A. muricata leaf extract was obtained
using supercritical fluid system and CO2
as solvent. A concentration of 2mg/ml
was made with 0.2% dimethyl sulfoxide
(DMSO) as diluent. The anti-angiogenic
activity of paclitaxel (4.6x10-2
mg/ml) and
bevacizumab (11.92 mg/mL) were
compared with A. muricata. All of these
concentrations were taken from prior
studies.
Fertilized duck eggs were incubated
for 10 days at 37°C with 70% humidity.
On Day 5, 2-3mL of albumin was taken
from the egg to decrease the pressure
inside and detach the embryo from the
shell. Egg windowing was done for better
manipulation of the sample. The window
was sealed with sterile tape and the eggs
were returned to the incubator for another
4 days. On Day 9, they were divided into
four groups with 0.2% DMSO as negative
control. They were inoculated with 20ul of
the each treatment and incubated for
another 24 hours. On Day 10,
angiogenesis evaluation was done using
KERN Optios stereomicroscope (10x) and
analyzed using WIMASIS image analysis
web-based system.
Antiangiogenic activity of !
Annona muricata (guyabano) extract
versus paclitaxel and bevacizumab!
1De La Salle Health Sciences Institute, 2De La Salle University - Manila, 3De La Salle - University Medical Centre!
Correspondence: marymanalo_md@yahoo.com!
Table 1 shows the total branching
points in the duck egg after the
administration of different treatments.
Duck embryos treated with 0.2% DMSO
yielded the highest mean with 158.4583
blood vessel branching points, thus
being the least antiangiogenic of the
treatments. This is followed by A.
muricata extract which exhibited
antiangiogenic activity by a mean of
117.7083 branching points. Third is the
chemotherapy drug, paclitaxel, which
yielded an average of 9.2917 branching
points. Lastly, the antiangiogenic drug
bevacizumab, yielded a mean of 2.5417
branching points, being the treatment
that inhibited the most number of
branching points.
These results indicated that the
optimal concentration of A. muricata
extract can inhibit angiogenesis but
comparative analysis of the mean
branching points presented graphically
in Figure 2 showed that it is not
comparable to the anti-angiogenic level
of paclitaxel and bevacizumab.
Post hoc comparison using the
Tukey- Kramer HSD test in Trial 1
showed a significant difference
between the antiangiogenic activity of
A. muricata leaf extract and other
treatments having a mean difference of
-157.83, 103, 111.17 with DMSO,
p a c l i t a x e l a n d b e v a c i z u m a b ,
respectively. In Trial 2, the mean
difference was -154.17, 105.67 and
113. In Trial 3, it resulted to -147.17,
109 and 116.17. Lastly, in Trial 4, it
yielded a mean difference of -137.50,
116 and 120.33. All these data showed a
significant difference at p<0.05. All of our
data favor paclitaxel and bevacizumab in
terms of their antiangiogenic activity.
Treatment
Mean branching
point
N
Standard
deviation
0.2% DMSO 158.46 24 44.52354
A. muricata 117.71 24 21.13410
Paclitaxel 9.29 24 8.30128
Bevacizumab 2.54 24 2.41335
DC
BA
Figure 2. Blood vessel of duck embryo treated with 0.2% DMSO (A),
A. muricata (B), paclitaxel (C) and bevacizumab (D).!
Cauilan CC1, Teves A1, Amil MT1, Condalor CA1, Orodio KM1,!
Santos VE1, Mendoza CW1, Durian A2, Macawile MC2, Gallo J2, Auresenia J2, and Manalo - Igot MOU3!
BACKGROUND RESULTS
METHODS
A B
Figure 1. Annona muricata fruit with leaves (A).
Annona muricata food supplements with
no approved therapeutic claims (B).
Table 1. Mean branching points of 10-day old duck
chorioallontoic membranes after treatment with A.
muricata leaf extract, paclitaxel and bevacizumab.!
CONCLUSION
Figure 3. Tukey-Kramer HSD test showing mean differences in blood
vessels treated with A. muricata leaf extract as compared
with 0.2% DMSO, paclitaxel and bevacizumab in four trials.!