S. Yogisha, K. A. Raveesha, /Journal of Natural Products, Vol.

3(2010):76-79
ISSN 0974 5211
Dipeptidyl Peptidase IV inhibitory activity of Mangifera indica
Shivanna Yogisha*, Koteshwara Anandarao Raveesha
Microbiology Laboratory, Department of Studies in Botany, Manasagangotri,
Mysore-570006, Karnataa, !ndia
"#orresponding aut$or
%&ecei'ed () *une (00+, &e'ised (6 *une-07 September (00+, -ccepted 0. September (00+/
AS!RA"!
0$e insulinotropic $ormone, glucagon-lie peptide 1 %2L3-1/, 4$ic$ $as been
proposed as a ne4 treatment for type ( diabetes, is metaboli5ed e6tremely by
Dipeptidyl peptidase !7 %D33-!7/8 !n$ibitors of D33 !7 en$ance t$e le'el of 2L3-1,
4$ic$ $a'e impro'ed glucose tolerance and increased insulin secretion8 0o understand
t$e t$erapeutic activity of Mangifera indica for t$e treatment of non-insulin dependent
diabetes mellitus %9!DDM/8 0$e met$anolic e6tract of Mangifera indica lea'es 4ere
tested in-vitro for D33!7 in$ibitory acti'ity8 0$e e6tract s$o4ed potent acti'ity 4it$ an
!#
50
'alue of 1.(87:g;ml8 Diprotin-- 4as used as reference standard8 0$e results
confirm t$e in$ibitory effect of M. indica on D33!7, and t$e potential to be a no'el,
efficient and tolerable approac$ for t$e diabetes8
Keywords# 2L3-1, D33!7, 9!DDM, Diprotin--, !n-'itro8
I$!R%D&"!I%$
- no'el approac$ for treatment of type- ( diabetes is based on t$e gut $ormone
glucagon-lie peptide-1 %2L3-1/, 4$ic$ is ant-diabetic due to its combined action to
stimulate insulin secretion, increase beta-cell mass, in$ibit glucagon secretion, reduce
t$e rate of gastric emptying and induces satiety8 0$e peptide is rapidly inacti'ated by
t$e en5yme dipeptidyl peptidase-!7 %D33-)/, resulting in a $alf-life of acti'e 2L3-1 of
only appro6imately 1-( minutes8
!n$ibition of D33-!7 increases t$e le'els of endogenous acti'e 2L3-1 and prolongs its
$alf-life8 0$e studies on animals s$o4ed genetic deletion of D33-!7, 4$ic$ $a'e
impro'ed glucose tolerance and increased insulin secretion in response to oral glucose8
&ecent studies in sub<ects 4it$ type ( diabetes $a'e s$o4n t$at prolonged D33-!7
in$ibition for up to 1 year is anti-diabetogenic because fasting and postprandial glucose
as 4ell as =b-1c le'els are reduced8 =ence, D33-!7 in$ibition $as t$e potential to be a
no'el, efficient and tolerable approac$ to treat type ( diabetes %Mentlein, 1+++ and Bo
-$r>n, (005/8
Mangifera indica L8 %?amily-Anacardiaceae/ is one of t$e most popular of all
tropical fruits8 Most parts of t$e tree @?ruit, seeds, pulp, stem bar, roots and lea'esA
$a'e medicinal properties %Sat$ya'at$i, et al8, 1+.7/8 !t is nati'e to tropical -sia and
#opyrig$t B (00+, *ournal of 9atural 3roducts, !9D!-, Dr8 Sud$ans$u 0i4ari, -ll rig$ts reser'ed
76
Journal of Natural Products
Volume 3 (2010)
www.JournalofNaturalProducts.com
Research Paper
S. Yogisha, K. A. Raveesha, /Journal of Natural Products, Vol. 3(2010):76-79
$as been culti'ated in t$e !ndian subcontinent for o'er )000 years and is no4 found
naturali5ed in most tropical countries8 Mangifera indica contains 'itamins - and #, C-
carotene, 6ant$op$ylls, $umulene, elemene, indicine, terpinine, tannins, fla'onoids,
linalool, nerol, gallic acid, et$yl gallate, met$yl gallate D mangiferin %-deribigbe, et al8,
1+++ and Eagner, 1++6/8
0$e lea'es of Mangifera indica 4ere assessed for antidiabetic properties using
normoglycaemic, glucose-induced $yperglycemia and strepto5otocin %S0F/ induced
diabetic mice %Gme5a4a, et al8, 1+.)/8 0$is e6periment designed to e'aluate t$e anti-
diabetic property of Mangifera indica lea'es on D33!7 in$ibitory acti'ity8 0$e aims of
t$is study 4ere to clarify met$anolic e6tract Mangifera indica lea'es in$ibits D33!7 to
determine in$ibitory acti'ities and to compare t$ese acti'ites in Diprotin, a reference
standard8 0$e present study e6plains one of t$e targets for diabetes and it $elps in
impro'ed glucose tolerance and insulin secretion8
'A!(RIA)S A$D '(!*%DS
Chemicals used# D33!7 from porcine idney, 2ly-pro-p-nitroanilide, Diprotein-- %!le-
pro-ile/, 0ris-=#l Buffer8 -ll c$emicals 4ere purc$ased from M;s Sigma, St Louis,
GS-8
Sample preparation: ?res$ lea'es of Mangifera indica L8 %-nacardiaceae/ 4ere
collected from Bangalore in 9o'ember (007 and t$e sample 4as aut$enticated
@&eference no8M!;3&H;0);07A8 0$e po4dered plant materials of 600 g 4ere e6tracted
4it$ met$anol using So6$let apparatus8 0$e resulting e6tracts 4ere e'aporated in
'acuum and finally lyop$ili5ed into solid mass de'oid of sol'ent %Iield J (0 K/ and
stored in desiccators for future use8
Dipeptidyl peptidase IV assays (in-vitro): 0$e assay 4as performed as per Ko<ima et
al %7/8 !n brief, t$e assay 4as performed in +6 micro 4ell plates8 - pre-incubation
'olume (50l contained 100mM 0ris =#l buffer p= .8), 785l of D33 !7 en5yme
%08(G;ml/ and 'arious concentration of test material;reference in$ibitor8 0$is mi6ture
4as incubated at L7# for L0mins, follo4ed by addition of 10l of 18) mM 2ly-pro-p-
nitroanilide %substrate/8 0$e reaction mi6ture 4as incubated for L0 mins at L7# and
absorbance 4as measured at )10nm8 Diprotein-- %!le-3ro-!le/ 4as used as reference
in$ibitor %7/8
Statistical Analysis: -ll data are e6pressed as t$e mean M SNM8 0$e Statistical data
4ere e'aluated by using 2rap$ pad 3rism) soft4are8 0$e K in$ibition 4as calculated
using t$e formula, control O test;control 6 1008 0$e !#
50
'alue 4as determined by non-
linear regression cur'e fit using 2rap$ pad 3rism)8
R(S&)!S
0$e met$anolic e6tract of Mangifera indica lea'es 4ere tested in Dipeptidyl
peptidase !7 in$ibitory assay %in-'itro/8 - rapid and simple micro dilution tec$niPue on
+6-4ell micro plate based on en5yme in$ibition mec$anism 4as optimi5ed and
'alidated for screening of antidiabteic acti'ity8
0$e $ig$est concentration tested for t$e Mangifera indica in D33!7 assay 4as
L(0:g;ml8 0$e 50K in$ibition 4as e6$ibited at 160:g;ml8 Mangifera indica lea'es
met$anolic e6tract in$ibited porcine idney D33-!7 4it$ an !#
50
of 1.(87:g;ml8 0$e
data pertaining to t$e antidaibetic potential of t$e Mangifera indica lea'es are presented
in 0able 1 D ?ig 1 respecti'ely8 0$e +5K confidence inter'al indicates t$e !#
50
'alue
4as found to be 4it$in t$e range of tested concentration8
#opyrig$t B (00+, *ournal of 9atural 3roducts, !9D!-, Dr8 Sud$ans$u 0i4ari, -ll rig$ts reser'ed
77
S. Yogisha, K. A. Raveesha, /Journal of Natural Products, Vol. 3(2010):76-79
!able+,# DPPIV inhibitory activity of Mangifera indica - Diprotin A.
S. $o. !ested /aterial "oncentration
0g1/l
2 inhibition 3
S.(.'
I"45 60g1/l7
6842 ".I7
a


1







Mangifera indica
%nJL/




0 181( M 1810
5 +8)7 M 18(0
10 1)8(( M 1850
(0 (78)) M 08)5
)0 L+855 M 08.0
.0 ).81( M 1810
160 )+8(( M 186)
L(0 6.8(( M 181)


1.(87
%.(8L5-(.)861/






(






Diprotin--

%nJL/
%!le-3ro-!le/
positi'e control


0 18(1 M 18(0
(85 1L875 M (8L)
5 ((871M 18L)
10 )187( M 1867
(0 6(857 M 187)
L0 678.. M 18.0
)0 7L8.L M 08+)


1+871
%+87-L(87+/




9ig:re+,
DIS"&SSI%$
2L3-1 is a substrate for t$e en5yme Dipeptidyl peptidase !7 %D33-!7/, a serine
protease 4$ic$ degrades 2L3-1 into its inacti'e form8 N6ogenous 2L3-1
administration $as been s$o4n to be useful in t$e treatment of type ( diabetes8
=o4e'er, t$e s$ort $alf-life maes 2L3-1 unattracti'e for c$ronic t$erapy of type (
diabetes8 D33-!7 in$ibition is an approac$ to prolong t$e circulating $alf-life of
2L3-1, t$us maing D33-!7 in$ibitors a promising target for t$e treatment of type (
diabetes8
Dipeptidyl peptidase-!7 %D33-!7/ is in'ol'ed in t$e inacti'ation of glucagon-
lie peptide-1 %2L3-1/, a potent insulinotropic peptide8 0$us, D33-!7 in$ibition can be
an effecti'e approac$ to treat type ( diabetes mellitus by potentiating insulin secretion
%Gme5a4a, et al8, 1+.) D -deribigbe, et al8, (001/80$is study describes t$e biological
effects of a natural plant e6tracts Mangifera indica in-vitro8 !n addition, t$e M. indica
met$anolic e6tract in$ibited D33-!7 mediated degradation of 2L3-1 in- vitro8 M.
#opyrig$t B (00+, *ournal of 9atural 3roducts, !9D!-, Dr8 Sud$ans$u 0i4ari, -ll rig$ts reser'ed
7.
S. Yogisha, K. A. Raveesha, /Journal of Natural Products, Vol. 3(2010):76-79
indica met$anolic e6tract e6$ibited competiti'e type of en5yme in$ibition8 0$e anti-
diabetic property of Mangifera indica lea'es maes t$is study uniPue %Eagner, 1++6/8
0$e result e6plains in$ibitory acti'ities on D33 !7 and may $a'e t$erapeutic potential
on type ( diabetes8
0$e present study underlines t$at Mangifera indica in$ibits t$e D33!7 and
en$ances t$e 2L3-1 for type ( diabetes8 0$is study demonstrates t$at Mangifera indica
met$anolic lea'es e6tract could be a good lead for furt$er de'elopment as a ne4 anti-
diabetic agent8
Ac;nowledge/ents# 0$e aut$ors are grateful to Department of Studies in Microbiology D
Botany for pro'iding facilities8
R(9(R($"(S
Mentlein, &8, %1+++/Q Dipeptidyl-peptidase !7 %#D(6/Q &ole in t$e inacti'ation of regulatory
peptides8 Regulatory Peptides, ()Q.5-.+8
Bo -$r>n8, %(005/Q !n$ibition of Dipeptidyl 3eptidase-) %D33-)/Q - 9o'el -pproac$ to treat
type (diabetes8 Current Enzyme Inhibition 1Q65-7L8
Gme5a4a, =8, -oyagi, =8, Hga4a, K8, %1+.)/Q Diprotein - and B, in$ibitors of Dipeptidyl
amino peptidase !7, produced by Bacteria8 he !ournal of Antibiotics" (6Q)((-)(58
Sat$ya'at$i, 2878, 2upta, -8K8, 0andon, 98, %1+.7/Q Medicinal plants of !ndia, !ndian
#ouncil of Medical &esearc$, 9e4 Del$i, !ndia8 pp8 (0+-(1(8
-deribigbe, -8H8, Nmudianug$e, 08S8, La4al, B8-8, %1+++/Q -nti$yperglycaemic effect of
Mangifera indica in rat8 Phytotherapy Research." 1LQ50)-5078
Eagner, =8, Bladt, S8, %1++6/Q 3lant Drug -nalysis, Springer 7erlag, (
nd
edition, 2ermany8
Ko<ima, K8, =am, 08, Kato, 08, %1+.0/Q &apid c$romatograp$ic purification of Dipeptidyl
peptidase !7 in $uman subma6iallry gland8 !ournal of Chromatography" 1.+Q(LL-()08
Gme5a4a, =8, -oyagi, =8, Hga4a, K8, %1+.)/Q Diprotein - and B, in$ibitors of Dipeptidyl
amino peptidase !7, produced by Bacteria8 he !ournal of Antibiotics" (6Q)((-)(58
-deribigbe, -8H8, Nmudianug$e, 08S8, La4al, B8-8, %(001/Q N'aluation of t$e antidiabetic
action of Mangifera indica in mice8 Phytotherapy Research." 5Q )56-)5.8
#opyrig$t B (00+, *ournal of 9atural 3roducts, !9D!-, Dr8 Sud$ans$u 0i4ari, -ll rig$ts reser'ed
7+