Received: 30 April 2021

| Revised: 31 May 2021

| Accepted: 6 June 2021

DOI: 10.1111/cbdd.13909

REVIEW

Alpha-­amylase as molecular target for treatment of diabetes

mellitus: A comprehensive review

Navjot Kaur | Vanktesh Kumar | Surendra Kumar Nayak | Pankaj Wadhwa |

Paranjit Kaur | Sanjeev Kumar Sahu

School of Pharmaceutical Sciences, Lovely

Professional University, Phagwara, Punjab,
Abstract
India The alpha (α)-­amylase is a calcium metalloenzyme that aids digestion by breaking
down polysaccharide molecules into smaller ones such as glucose and maltose. In
Correspondence
Sanjeev Kumar Sahu, Lovely Professional addition, the enzyme causes postprandial hyperglycaemia and blood glucose levels
University, Department of Pharmaceutical to rise. α-­Amylase is a well-­known therapeutic target for the treatment and main-
Sciences, Phagwara, Punjab, 144411 India.
tenance of postprandial blood glucose elevations. Various enzymatic inhibitors,
Email: [email protected]
such as acarbose, miglitol and voglibose, have been found to be effective in target-
ing this enzyme, prompting researchers to express an interest in developing potent
alpha-­amylase inhibitor molecules. The review mainly focused on designing differ-
ent derivatives of drug molecules such as benzofuran hydrazone, indole hydrazone,
spiroindolone, benzotriazoles, 1,3-­diaryl-­3-­(arylamino) propan-­1-­one, oxadiazole
and flavonoids along with their target–­receptor interactions, IC50 values and other
biological activities.

KEYWORDS
alpha-­amylase, alpha-­amylase inhibitors, antidiabetic drug, diabetes mellitus, molecular target

1 | IN T RO D U C T ION different types of amylase enzymes, one major category of

enzyme is pancreatic α-­amylase. These are basically calcium
As per WHO, the worldwide estimation of global pervasive- metalloenzymes (Samrot & Vijay, 2008) due to which cal-
ness for diabetes in 2019 was 463 million and it is expected to cium is an important cofactor in performing functions such
increase by 700 million in 2045. Diabetes mellitus is a met- as digestion of starch (Morris et al., 2011). Carbohydrates
abolic disease consisting of chronic hyperglycaemia, which when consumed firstly should break down into smaller frag-
results from defects in the secretion of insulin, abnormal in- ments such as monosaccharides, which would further absorb
sulin action or sometimes both. The critical symptoms of di- in the body. α-­Amylase engages the catalysing of α-­(1,4)-­
abetes depend upon type and duration of it given in Figure 1. D-­glycosidic linkages present in starch to hydrolyse them
Untreated patients may suffer from stupor, numbness, into smaller fragments and other polymers of glucose. By
coma and even death because of ketoacidosis (Kahanovitz performing the act of inhibiting α-­amylase enzyme, it helps
et al., 2017; Kharroubi & Darwish, 2015). in reducing hyperglycaemia, obesity and problems such as
Amylase is an enzyme produced by salivary glands overweight conditions. The polyphenolic inhibitors (diet
that digests starch molecules to give a breakdown product usually obtained from plants and fruits), proteinaceous and
such as maltose, which in turn cleaves into two molecules pseudosaccharides have also been reported for decreasing
of glucose. To continue digestion of incoming starch, the blood glucose levels (Mahmood, 2016). Salivary alpha-­
pancreatic duct sends a heavy amount of pancreatic amylase amylase was first named as ptyalin, an agent present in saliva
to the duodenum. Amylase serves both endocrine and exo- for disintegration of starch content in food. In recent times,
crine functions (Des Gachons & Breslin, 2016). Among the a plenty of information has been gathered and established

Chem Biol Drug Des. 2021;00:1–22. wileyonlinelibrary.com/journal/cbdd © 2021 John Wiley & Sons Ltd | 1
2
|    KAUR et al.

about the enzyme produced in saliva along with its interac- diabetes of the young) (Urbanová et al., 2018) and neonatal
tion towards carbohydrates. In humans, gene code AMY1 diabetes (Lemelman et al., 2018).
is denoted as salivary gene, which helps in the production Type 1 diabetes (T1D)/juvenile diabetes is an autoimmune
of enzyme salivary alpha-­amylase in the saliva. In chromo- disease where the body fails to produce insulin. Patients are
some 1, AMY1 is present on the shorter arm (Butterworth insulin-­dependent, and they must have to take insulin artifi-
et al., 2011; Smith et al. 2010). As α-­amylase is an important cially to live. Type 2 diabetes is defined as the type of dia-
target for the treatment of diabetes mellitus and development betes where it gets resistant to insulin due to its insensitivity;
of new drugs, scientists are showing their great interest in therefore, insulin production decreases, which leads to fail-
this enzyme. This review mainly focused on all the possi- ure of pancreatic beta-­cell and glucose transport to muscle
ble alpha-­amylase inhibitors reported from 2000 to till now. cells, liver and fat cells (Blair, 2016). Gestational diabetes
Currently, Acarbose and Miglitol are widely used; however, is a situation in which blood sugar level of a woman become
there are some side-­effects such as flatulence, diarrhoea, high during pregnancy. In such case, a woman without di-
bloating and abdominal discomfort (Hemlata et al., 2019). abetes cannot able to make and use all the insulin after the
Generally, carbohydrates are an immense source of energy pregnancy.
in human body with almost 40% to 80% of its portion in
a meal. They are mainly classified into four major cate-
gories according to the chemical structure and behaviour. 2 | POSSIBLE TARGETS FOR
These are non-­fermentable, fermentable, digestible and un- ANTIDIABETIC DRUGS
digested (absorbable). There is no requirement of digesting
some carbohydrates (monosaccharide) known as absorbable There are various targets that are affected by different
carbohydrates to provide the needed energy. They directly chemical agents to induce their antidiabetic effect (Table 1).
get absorbed into the body. All the higher units other than Synthetic derivatives acting on various novel protein tar-
monosaccharide must be digested; otherwise, they can get gets to treat diabetes may be classified on the basis of their
accumulated into the gut or ileum or intestine or colon (with- mode of action as insulinotropic or non-­insulinotropic agents
out fermentation) and remain undigested when administered (Kneuer et al., 2006; Tiwari et al. 2014).
with alpha-­amylase inhibitors or alpha-­glucose inhibitors.
The side-­effects such as GI distress, bloating and abdominal
pain can occur with these conditions (Jain et al., 1989; Tucci 3 | ALPHA-­A M YLASE AS
et al., 2010). However, taking 50% of gallic acid along with ANTIDIABETIC M OLECULAR
50% of alpha-­amylase inhibitor (acarbose) can reduce the TARGET
side-­effects associated with it, mainly oxidative stress (Oboh
et al., 2016). α-­Amylase is the starch hydrolase enzyme with various se-
The diabetes is categorized into three major categories quences of amino acids present in them. To yield glucose
as (i) type 1 diabetes or insulin-­dependent diabetes or ju- and maltose, α-­amylases cleave the α-­1,4 glycosidic link-
venile diabetes; (ii) type 2 diabetes or insulin-­independent ages in amylose. α-­Amylase is also named as Taka-­amylase
diabetes; and (iii) gestational diabetes. Further, several A, after the name of discoverer Takamine (Kitamoto, 2002).
studies have specified the diabetes on different bases such α-­Amylase enzyme is the major digestive enzyme in sa-
as steroid-­induced diabetes (Hwang & Weiss, 2014), double liva. The α-­1,4 glycosidic linkages are hydrolysed in starch
diabetes (Merger et al., 2016), brittle diabetes (Vantyghem & groups. Gastric acid in the stomach makes α-­amylase inac-
Press, 2006), LADA (latent autoimmune diabetes of adult- tive. That is why α-­amylase works best at a slightly alkaline
hood) (Pozzilli & Pieralice, 2018), MODY (maturity-­onset pH (Smith et al. 2010).

FIGURE 1 Symptoms of diabetes

KAUR et al.   
| 3

3.1 | Types of amylase enzymes hydrolyse molecules of starch to give different products such
as dextrin and glucose units (in a small polymer), which
There are different types of amylase enzymes mentioned in causes hyperglycaemia and development of T2D (Shankaraia
Figure 2, but out of them, α-­, β-­and γ-­amylase enzymes play & Reddy, 2011). This pancreatic α-­amylase involves hydrol-
important biological role in carbohydrate metabolism (Saini ysis of the α-­1,4 glycosidic linkages of the glycogen, amy-
et al., 2017). α-­Amylases are enzymes that have ability to lopectin starch, amylose, dextrin and maltose in body and is

TABLE 1 Different targets for anti-­diabetic agents with their mechanism with examples

Targets Mechanism of action Marketed drugs Drugs in clinical trials

Protein Tyrosine Inhibiting PTP1B Norathyroil O
OH
NH2 H3C O S
Phosphatase1 B increases “insulin H OH O
CH3 H
(PTP1B) (Goldstein, sensitizing”. CH3
O CH3 H
2001) H HN H3C
HN
HO H H
N OH
H H

MSI-1436
Erybraedin A

Dipeptidyl peptidase-­4 DPP-­4 inhibitors FF NH

F
(DPP-­4) (Crepaldi et al., control N
N S
F N
2007) hyperglycaemia by N N N O
N N N
stimulating insulin
NH2O
secretion. F
F Teneligliptin (MP-513)
Sitagliptin,
N
N

HO NH O

Vildagliptin

Free fatty acid receptors Increases insulin O

1 (FFAR1) activation secretion during OH
O
(Eleazu et al., 2018) increased glucose N S
HOOC
levels. O

α linolenic acid LY2922470

G protein-­coupled Inhibition decreases O HO O

receptors (GPCR) glucose-­level. OH

N N
O
H
(Sebastiani et al., 2018) O
N
H NH O
NH O
OH
Oleoylethanolamide, NHO
O NH2
O NH
OH NH O
H
O O O R HO N
N+ P N
H
O OH O
O O
NH

N NH2
H
Lysophosphatidylcholi
ne Ghrelin (azp-531)

Peroxisome Proliferator Enhance insulin S O O

activated receptor-­γ action by increasing NH
N O O
(PPARγ) (Derosa & uptake of glucose. O
O
Maffioli, 2012) OH O
Pioglitazone
O
Naveglitazar

(Continues)
4
|    KAUR et al.

TABLE 1 (Continued)

Targets Mechanism of action Marketed drugs Drugs in clinical trials

α-­glucosidase inhibitors Inhibit carbohydrate HO OH

(A GIs) (van de Laar, absorption from the HO O OH

2008) gut. O
O OH
HO
O OH
OH
HO
O OH

HN OH

HO OH

Acarbose
OH
OH
HO N

HO
HO

Miglitol

Aldose reductase Blocks glucose O O

inhibitors (Tomlinson breakdown by polyol N N
N F
et al., 1994) pathway. N S F
F N
O F
HN OH
NH O
O AT-001
Sorbinil
O
O
NH2

F O
NH
HN
O
Fidarestat
O F F
F
OH

N N
N
S
O

Zopolrestat

Glycogen phosphorylase Attenuate

N
inhibitors(GP) (Baker hyperglycaemia OH O O
HO O H H O
et al., 2005; L Kantsadi HO
N N H 2N N HO
HO H
et al., 2015) OH
O O O
Efpeglenatide
N-(3,5-Dimethyl-
Benzoyl)-N-(β-D-
Glucopyranosyl)urea.

(Continues)
KAUR et al.   
| 5

TABLE 1 (Continued)

Targets Mechanism of action Marketed drugs Drugs in clinical trials

Fructose-­1,6-­ Control O
O
bisphosphatase inhibitor gluconeogenesis HO H 2N N O P CO2Et
HO O N
(FBPase) (Kaur et al., (overproduction) HN H
2017) OH S
OH CO2Et
HO
O

Achyrofuran
MB06322 (CS-917)

Glucagon receptor Suppression of Cl F O O

Cl F
inhibitor (GCGr) (Lee glucagon F N OH
et al., 2016)
O H
O
N
H
N
O
O
O
O OH
N

LY2409021
Albiglutide

Phosphoenol pyruvate Control hepatic —­

carboxykinase (PEPCK) gluconeogenesis O
(Wall et al., 2015) HN O N
HN S
O NH N

Glipizide

Sodium glucose co-­ Inhibits secretion or O Cl —­

transporters (C Rizos & utilization of insulin O
OH
S Elisaf, 2014)
HO OH
OH

Dapagliflozin

O F
HO S

HO OH
OH

Canagliflozin
HO

OH
O
O OH
OH
O Cl

Empagliflozin

α-­amylase inhibitors Regulates blood HO OH —­

(Agarwal & Gupta, glucose and
HO O OH
2016) postprandial glucose O
levels O OH
HO
O OH
OH HO
O OH

HN OH

HO OH

Acarbose
6
|    KAUR et al.

effectively responsible for digestion of starch. Starch being (mashing) industry (Fox, 2018; Toldra & Kim, 2016). γ-­
a big molecule cannot cross the blood–­brain barrier. Thus, Amylase is an enzyme mainly found in plant and animal
α-­amylase cleaves the starch into smaller glucose unit as it tissues. To produce glucose from amylopectin or amylose,
is needed by brain. Excessive conversion can increase the γ-­amylase hydrolyses both the glycosidic linkages α-­1,4 and
blood sugar level in tissues, and due to overactivation of α-­ α-­1,6-­glucosidase (Mojsov, 2016).
amylase and deficiency of insulin, hyperglycaemia condition
can occur in some cases (Agarwal, 2016).
β-­
Amylase is a type of amylolytic enzymes (Zhang 3.2 | Role of α-­amylase inhibitors in
et al. 2010) or maltogenic amylase (Eck, 2013), which helps treatment of diabetes mellitus
in the breakdown of α-­1,4-­glucan linkage at 4.0–­5.5 pH by
hydrolysing it into maltose. The ‘beta’ term is used due to Molecules such as starch are large that is the reason they
anomeric configuration of freely available maltose groups cannot cross the blood–­brain barrier. But on the other side,
released initially and not due to hydrolysis of α-­1,4-­glucan glucose is required for the proper brain function. Hence, α-­
linkage. It is mainly found in the seeds of higher plants and amylase enzyme converts the large and complex molecule
microbes such as bacteria and fungi, and responsible for the of starch into small sugar fragments so as to make them
sweet taste of fruits. β-­Amylase can easily be seen in sweet easy to cross the BBB. When excess of the starch starts
potatoes and ripened fruits. The enzyme is used in the forma- converting into simpler sugar fragments, it increases the
tion of syrups high in maltose, and in distilling and brewing blood glucose levels. Postprandial glucose levels are also
increased by α-­amylase. This is the reason α-­amylase inhib-
itors are being used, and its mechanism is given in Figure 3.
α-Amylase Acarbose is an antidiabetic agent used for suppressing α-­
amylase enzyme (Agarwal, 2016). Our area of interest is
Endo- β-Amylase α-­amylase inhibitors, and we are going to survey all the
Amylase literature available about it so as to gather most of the in-
Amylase
formation. These inhibitors are described as good targets
for the treatment of disorders such as diabetes and obesity
Exo- γ-Amylase (Sales et al., 2012). This inhibitor makes an environment
Amylase in body, so that there is a delay in the breakdown of carbo-
hydrate, and reduces the postprandial blood glucose levels
(Kazeem et al., 2013).
FIGURE 2 Types of amylase enzymes

FIGURE 3 Mechanism of action α-­amylase inhibitors

KAUR et al.   
| 7

3.2.1 | Flavonoids as α-­amylase inhibitors flavonols, flavanols, isoflavones and flavones) are different
from one another mainly in the type of heterocycle of ring C,
Flavonoids play an important role in medicinal chemistry. and substitution of –­OCH3 and -­OH groups at the different
Thus, investigation and testing of twenty-­one flavonoid com- positions (R3, R5, R6 and R7) on ring A-­C and on ring B
pounds was done. Inhibitory activities of all were examined (R3′ and R4′). Only a few compounds of flavonol and fla-
against α-­amylase and α-­glucosidase. Luteolin has α-­amylase vone families showed promising results of amylase inhibition
inhibitory activity, but its potency was lower than acarbose. (Lo Piparo et al., 2008).
Different clinical tests and further evaluation were needed. A medicinal plant named as Vaccinium arctostaphylos has
Flavonol glycosides were found to have a α-­amylase inhibi- been mentioned for the treatment of diabetes mellitus by ex-
tory activity (Kim et al., 2000). amining the IC50 values (Table 4). The extract obtained from
As per the reported data by Matsui et al., the extraction its berries showed an inhibitory effect on α-­amylase in vitro.
of twelve anthocyanins was carried out and they were found Hence, the extract was purified, which leads to isolation of
to have a potent inhibitory activity towards α-­glucosidase an α-­amylase inhibitor, that is malvidin-­3-­O-­β-­glucosides,
(AGH). Extract of Pharbitis nil (SOA) was found to have the reported in Figure 5 (Kianbakht et al., 2013).
strongest inhibitory activity towards maltase as similar to the A dose-­ dependent reduction in the α-­amylase activity
extract of Ipomoea batata (YGM) (Table 2). [IC50 = 1910 (1,890–­1940) µg/ml] was observed from extract
Both extracts were reported to show inhibition of α-­ obtained from V. arctostaphylos berries. This was the first
amylase action and indicated the good characteristic of report showing the inhibitory effect produced by malvidin-­
suppressing postprandial blood glucose levels (Matsui 3-­O-­β-­glucoside on α-­amylase activity (Sudha et al., 2011).
et al., 2001). In a study, McCue et al. treated porcine pancre-
atic α-­amylase (PPA) herbal extracts containing rosmarinic
acid (RA). Activity for α-­amylase inhibition was noticed via
these in vitro experiments where phytochemicals were uti- R3'
lized. The results indicated that α-­amylase inhibitory activity
R4'
increases with increasing concentration of RA (P. P. McCue
& Shetty, 2004).
Structural requirements of flavonoids for management of R7 O
R5'
postprandial glycaemia and starch digestion were evaluated.
Out of 19 flavonoids, three flavones and four flavonols were R6'
found exhibiting IC50 values lower than 100 μm (Table 3 & R6
Figure 4). The five subclasses of flavonoids (flavanones, R5 O

FIGURE 4 General structure of flavonoids

T A B L E 2 Pharbitis nil (SOA) extract and Ipomoea batata
(YGM) extract with their IC50 value
T A B L E 4 Inhibitory activities for α-­amylase and IC50 values of
IC50 value the berry extract of Vaccinium arctostaphylos along with its active
Extracts (µg/ml) compound Mv-­3-­glc
Pharbitis nil (SOA) extract 350 µg/ml
Inhibition
Ipomoea batata (YGM) extract 360 µg/ml Concentration (%) IC50
Berry extract [µg/ml]
TABLE 3 Experimental maximum inhibition and IC50 values
3,600 84.95 1910 (1,890–­1,940) µg/ml
IC50 Maximum 2.880 74.97
Compound Family (μm) inhibition (%) 2.300 60.96
Acarbose Polyol 0.996 99.2 1,840 36.66
Quercetagetin Flavonol 10.2 97.6 1,470 19.48
Fisetin Flavonol 19.6 85.6 Mv-­3-­glc [μm] 0.329 (0.316–­0.342) μm
Quercetin Flavonol 21.4 82.1 0.913 72.60
Myricetin Flavonol 30.2 79.0 0.584 55.15
Scutellarein Flavone 9.64 98.4 0.374 44.05
Luteolin Flavone 18.4 88.8 0.239 21.79
Eupafolin Flavone 48.0 99.4 0.153 8.75
8
|    KAUR et al.

OCH3 was done for migration of acyl part of acteoside to isoacte-

OH oside (Lu et al., 2017). Their chemical structures and per-
2'
centage composition are reported in Figure 10 and Table 6,
HO 8 O+
OCH3 respectively.
6'
HO 2''
3'' OH
Conformational changes were observed when bind-
6
O 4'' OH ing of isoacteoside and acteoside to α-­amylase took place.
4 1'' O
OH 5'' 6'' OH Decreased α-­ amylase activity was observed because the
binding site of isoacteoside and acteoside was near to the
FIGURE 5 Chemical structure of malvidin-­3-­O-­β-­glucoside active site region of α-­amylase. In vitro α-­amylase inhibi-
tory activity was examined with preheated acteoside, which
Arcytophyllum thymifolium belongs to family Rubiaceae is shown in Table 7. Isoacteosides were less efficient and
is a flowering plant. Increasing order of polarity was taken effective than acteosides for α-­amylase inhibitory activity.
into consideration while choosing the solvent for extraction The main reasons are as follows: firstly, the interactions be-
of the aerial parts, thereby six new (three new coumarin com- tween α-­amylase and acteoside were more potent than inter-
pounds (1–­3), a prenylated flavanone (4) and two iridoids action between α-­amylase and isoacteoside; and secondly,
(5 and 6) and (7–­23)) and 17 known secondary metabolites it was reported in the docking studies that binding site of
were obtained, where only (1, 2, 7, 10, 13–­15, 18, 20 and isoacteoside was much more away as compared to binding
23) compounds showed a better IC50 value with different site of acteoside.
potentials. The chemical structures of all these identified In 2018, Gonzalez et al. evaluated the binding capacity
phytoconstituents are mentioned in Figure 6. Compound 13, of different flavonoids with pancreatic alpha-­amylase. These
that is rhamnetin (73.9 μm), and compound 15, that is aspe- flavonoids are quercetin (QUE), hesperetin (HES), catechin
rulosidic acid (69.4 μm), were found to have α-­amylase in- (CAT), luteolin (LUT) and rutin (RUT). By using techniques
hibitory effect comparable with acarbose (26.3 μm), which such as molecular docking, UV-­Visible spectroscopy and flo-
is shown in Figure 7 (Balan et al., 2016). The α-­amylase rescence, the resultant potent activity was examined. Luteolin
inhibitory activity of potent secondary metabolites obtained in comparison with the reference acarbose (ACA) was shown
from Arcytophyllum thymifolium is also reported in Figure 8 to have the better inhibitory action. Hence, alpha-­amylase had
graphically. a high affinity towards LUT. On the other hand, rutin (RUT)
Salacinol derivatives and their sulphur analogues were and catechin (CAT) were shown to have no inhibitory activity.
introduced as potential inhibitors of both intestinal and Docking and fluorescence analyses evaluated that interactions
pancreatic α-­amylase to slow down production of glucose between enzyme and flavonoid were controlled by the hydro-
from digestion of food (Table 5). Thus, these agents may phobic interactions. The activity of pancreatic α-­amylase was
provide an opportunity to develop them as alternate for measured in the absence and presence of flavonoids, HES,
reduction of blood glucose levels in T2D. The maltase glu- LUT, QUE, CAT and RUT, and the positive control ACA. The
coamylase (MGA) enzyme is involved in the final step of enzyme–­flavonoid (CAT, LUT, RUT and QUE) interactions
digestion of starch and release of glucose. Salacinol (a nat- were evaluated in the presence and absence of flavonoids.
urally occurring α-­glucosidase inhibitor), its derivatives The chemical structures of different flavonoids are shown in
and acarbose (antidiabetic agent) have been evaluated as Table 8 with their inhibitory activity. In silico docking studies
inhibitors of MGA through their binding to N-­terminal showed that Asp197, Glu233 and Asp300 amino acids were
catalytic domain of enzyme. The five salacinol derivatives closely bound in enzyme–­flavonoid interactions (Martinez-­
(1–­5) were supposed to have better inhibitory activity Gonzalez et al., 2019).
than acarbose and their structures, which is described in In the same year, to check structure–­activity relationship
Figure 9. Further, replacement of selenium with sulphur between 40 different flavonoid compounds, (Proenca et at.,
in salacinol derivatives in 5-­membered ring resulted in sa- 2019) conducted an evaluation test through kinetic analysis
lacinol analogues with variable MGA inhibitory activities and molecular docking. It was found that position and nature
(Rossi et al., 2006). of flavonoid substituents play a vital role in alpha-­amylase
Worst inhibition was seen in acarbose (Ki = 62 μM), enzyme inhibition. The positions 5 and 7 of A-­ring, positions
while best inhibition was seen in salacinol and compound (5) 30 and 40 of B-­ring that consist of –­OH group, and position
against MGA (Ki = 0.2 μM). 3 that consists of –­Cl made the flavonoid best for competitive
Lu et al. worked on acteoside, the polyphenol obtained amylase inhibition among all the 40 compounds. IC50 value
from small leaves of Kudingcha (Ligustrum purpurascens) of D11 is 44 ± 3 µm, and its chemical structure is reported in
(the Chinese tea) was found to exhibit enormous biologi- Figure 11 where the reference compound acarbose consists
cal activities. Treatment of acteoside with high temperature of 1.3 ± 0.2 µm.
KAUR et al.   
| 9

OCH
OH
R O 3 OCH
1 3
OH

O O O HO O O HO O O H3C O O

(1) R1= CH3 HO HO CH3

(3)
HOH C
2 (2) R1= H (7)
OH
H3C OHCH3 H3C CH2 (4) OH O

R
COOR 3
1
R OH
2
H3CO
R O O
O 1

R OH C HO O O
3 2
OH OR
2
(11)
HOH C OH O
2 HOH C
O HO 2
O
HO O
(12) R1= H R2= CH3
HO OH
O R3= OH
OH HO OH
(5) R1= CH3 R2= OH R3=
H3C (13)R1= CH3 R2= H R3= OH
(15) R1= H R2 = OH R3= COCH3 (14)R1= R3= H R2= HO HOH C
2
O
(16) R1= CH3 R2= OH R3= H CH3
(17) R1= CH3 R2= OH R3= COCH3 HO
(18) R1= R3= H R2= OH O HO HOH C
2 O
(21) R1= CH3 R2= OH R3= OH O
H3C HO OH

HO COOR 1

H CO O OR2 OR3
3
O
O OH

O O O
(22) R1= CH3 R2=
(9) O
H3C OH
O
H3C OH OH
R1OH2C
HOH C
(23) R1= R2 = H R3=
HO 2 O
O
HO OH
OH
OH
O
(6) R1 = OH
H3C

(19) R1 = H
(20) R1 = COCH3

F I G U R E 6 Different compounds with structure-­hedyotiscone A (8),toddanin (9), hedyotiscone B (10),fraxetin (11), quercetin 3-­O-­β-­D-­
galactopyranoside (12), rhamnetin (13), kaempferol 3-­O-­β-­D-­glucopyranosyl-­(1→2)-­β-­D-­galactopyranoside (14), asperulosidic acid (15), deacetyl
asperulosidic acid methyl ester (16), asperulosidic acid methyl ester (17), scandoside (18), deacetyl asperuloside (19), asperuloside (20), 10-­O-­p-­
hydroxybenzoyl scandoside methyl ester (21), neochlorogenic acid methyl ester (22) and chlorogenic acid (23).

3.2.2 | Acarviostatins as α-­amylase inhibitors therapeutic agent for diabetes. Until the date, Acarviostatin
III03 was the most potent known inhibitor. The oligomer was
ZG0656 stain is a novel variety of S. coelicoflavus with thought to be beneficial for finding connection of α-­amylase
good α-­amylase inhibitory activity. Acarviostatins III03 was with other inhibitors so as to get more choices for diabetes
found to show the most significant effect of lowering glucose treatment. Four chemically different α-­ amylase inhibitors
levels in blood and would be developed towards a possible were isolated from the filtrate obtained with help of strain
 10
|    KAUR et al.

O OH F I G U R E 7 Structure of rhamnetin (13)

HO and asperulosidic acid (15)
H
O
OH O
OH H3C O OH
H
O OH
H3C O O H
O
OH
OH
O HO
OH
(15)
(13)

F I G U R E 8 α-­Amylase inhibition
by extract obtained from Arcytophyllum
thymifolium
IC50 (μM)

TABLE 5 α-­Amylase inhibitors and their IC50 values was regarded as potent target for T2D. Different acarvios-
tatins obtained from Streptomyces coelicoflavus var. nan-
Inhibitor Ki (μm)
kaiensis formerly exhibited more potency for inhibiting
Acarbose 62.0 ± 13 pancreatic amylase (Table 9). Here, the acarviostatin in-
Salacinol (1) 0.19 ± 0.02 hibitor series (I03, II03, III03 and IV03) were complexed
2 0.49 ± 0.05 with HPA crystal structures. Acarviostatin I03 showed
3 0.26 ± 0.02 comparable similarities to acarbose in condensation re-
4 0.25 ± 0.02 actions and hydrolysis at HPA active sites. Hydrolysis re-
actions are shown by acarviostatin (II03, III03 and IV03)
5 0.17 ± 0.03
only. By combining structural analysis and kinetic assays,
it was demonstrated that the best-­fitted structures for oc-
cupying active site fully were found to have seven sugar
ZG0656 in culture. These inhibitors were named as acarvio- rings. These structures were found to show the most ef-
statins I03, II03, III03 and IV03. This strain was obtained fective inhibitory activity. The interaction between subsite
from a bacterium species S. coelicoflavus (Geng et al., 2009). 4 of the HPA active site and inhibitor is examined with
By incubation with porcine pancreas α-­amylase (PPA), pro- high-­resolution structures. These results provided import-
cessing products of acarbose, acarviostatin I03, acarviostatin ant information for treating obesity or T2D by designing
II03 and acarviostatin III03 were obtained. Their chemical new drugs (Qin et al., 2011).
structures are also shown in Figure 12. Interactions between AII03 (14.7), AIII03 (14.3) and
Again Qin et al. found that the D-­(1,4)-­glycosidic link- AIV03 (41.6 µM) occurring at subsite 4 of HPA were ob-
age was catalysed by human pancreatic α-­amylase (HPA), served as 50–­200 times better than interactions with acarbose
which hydrolysis it into starch. This is the reason that HPA (Qin et al., 2011).
KAUR et al.   
| 11

F I G U R E 9 Compounds with OH OH OH
complete α-­amylase inhibitory activity are
(1) salacinol, and compounds (2–­5) are OH
derivatives of salacinol having variations OH
in the length and stereochemistry of the S OSO3 S OSO3
aliphatic chain (Rossi et al., 2006)
HO HO

HO OH HO OH
(1) (2)

OH OH OH

OH OH
Se OSO3 S OSO3 OH
HO HO

HO OH HO OH
(3) (4)

OH OH

OH
S OSO3 OH
HO

HO OH
(5)

OH T A B L E 6 Composition of acyl-­migrated acteoside (%) at

OH respective temperatures
HO O O
O
OO OH Temperature Isoacteoside Acteoside
H3C
HO OH
HO O 100°C 10.21 87.79
OH 120°C 21.73 78.27
OH
Acteoside

HO O
O OH T A B L E 7 α-­Amylase inhibitory effect of isoacteoside-­preheated
HO O O
HO acteoside and acteoside
OH
O
H3C OH Compounds IC50 (µm)
HO O
Isoacteoside 510.39 ± 8.42
OH
OH Acteoside 100°C 213.21 ± 6.10
Isoacteoside Acteoside 120°C 257.86 ± 9.71
Acteoside 200.46 ± 7.87
FIGURE 10 Chemical structures of acteoside and isoacteoside
12
|    KAUR et al.

T A B L E 8 Chemical structures of
Name Chemical structure IC50 (µm)
different flavonoids along with their
Luteolin (LUT) OH 18.00 ± 1.00 inhibitory activity
OH

HO O

OH O
Quercetin (QUE) OH 12.70 ± 1.20
OH

HO O

OH
OH O

Hesperetin (HES) OH 20.10 ± 0.70

O

HO O

OH O
Catechin (CAT) OH Not
determined
HO O
OH

OH
OH
Rutin (RUT) OH Not
determined
HO O
OH
O OH
O O
O
OH O OH
OH OH OH
OH
Acarbose (ACA) OH 14.60 ± 1.70
HO OH

OH
HN
OH

O
OH OH
HO O

OH
O O
OH
HO
O
OH
OH
KAUR et al.   
| 13

3.2.3 | Indole and benzofuran hydrazone activity. It worked as α-­glucosidase and α-­amylase inhibitor
analogues as α-­amylase inhibitors (Altowyan et al., 2019). All these analogues are reported in
Table 11 with their chemical structures and IC50 values.
Synthesis of twenty indole hydrazone analogues was car- Four of the analogues were found to show potent inhibi-
ried out. After investigation, IC50 values in between 1.66 tory activity for α-­glucosidase and α-­amylase. All other ones
and 2.65μm were observed for the derivatives (Table 10). It were found to show mild-­to-­moderate effective inhibitors of
has been reported that nine compounds were found to show α-­amylase (Altowyan et al., 2019).
potent inhibitory effects when comparative studies were car- Taha and Syahrul have synthesized benzofuran hydrazone
ried out by taking acarbose as standard. All other compounds derivatives and carried out in vitro study for examining potency
were found to show good-­to-­moderate potency towards α-­ of α-­amylase inhibitors with IC50 values (1.245–­2.320 μm)
amylase. These nine compounds with their IC50 values and (Taha et al., 2017). The IC50 values and their chemical struc-
chemical structure are reported in Table 10 and Figure 13, tures are reported in Table 12 and Figure 14, respectively.
respectively (Noreen et al., 2017). Nine of the derivatives have been reported for their potent
In 2019, Altowyan et al. have reported eighteen spiroindo- inhibitory activity, while others were found to have good-­
lone analogues, which assured to have potent hypoglycaemic to-­moderate activity against α-­amylase when acarbose was
taken as a standard (Taha et al., 2017).
OH
3.2.4 | Benzotriazoles as α-­amylase inhibitors

Hameed et al. in the same year synthesized 34 benzotria-

HO O zole derivatives with good-­ to-­
moderate α-­ amylase and
α-­glucosidase inhibitory activity with IC50 values of 2.00–­
5.72 and 2.04–­5.60 μM, respectively, which is shown in
Cl OH Table 13. Out of thirty-­four, eight derivatives were car-
rying potent inhibitory activity (Hameed et al., 2019) and
OH O basic chemical nucleus for all these derivatives is shown in
Figure 15.
FIGURE 11 Flavonoid derivative (D11)

FIGURE 12 Chemical Structures of acarbose, acarviostatin I03, acarviostatin II03 and acarviostatin III03
14
|    KAUR et al.

TABLE 9 Acarvostatins and acarbose with their inhibition TABLE 11 Spiroindolone analogues and their IC50 value
constants
α-­amylase
Inhibitors Inhibition potency Ki (μm) Spiroindolone analogues IC50 value (μm) selectivity
Acarviostatin I03 2.07 1.2549 ± 0.0350 Cl 39.02 ± 1.73 0.74
Acarviostatin II03 176.81 0.0147 ± 0.0012
Acarviostatin III03 181.34 0.0143 ± 0.0002 N
Acarviostatin IV03 62.39 0.0416 ± 0.0098 HN
O
Acarbose 1.00 2.5932 ± 0.0125 O
O

T A B L E 1 0 Indole hydrazone derivatives with potent α-­amylase

inhibitory activity F
F F
A IC50 (μm)

3C O 2.23 Br 49.28 ± 1.09 0.79

HO OH 2.44 N
HN
O
O
Br
1.92 O

O – 2.49
N+ O
F
F F
N
1.66
Cl 37.22 ± 1.49 0.70
2.25
N
O 1.87 HN
– O
O
N+ O
O O

H 1.97
N
N NH2
H 1.83 22.61 ± 0.54 0.62
O N

N
Acarbose 1.05 HN
O
O
O
H
N
NH2
N NH Standard acarbose 0.75 ± 0.07 3.31
CH3 A Note: Four of the analogues were found to show potent inhibitory activity for α-­
O glucosidase and α-­amylase. All other ones were found to show mild-­to-­moderate
effective inhibitors of α-­amylase (60).
FIGURE 13 3-­Methylindole hydrazone derivative

3.2.5 | Arylamines and arylimines as α-­ series of molecules with α-­amylase inhibitory activity (Bashary
amylase inhibitors & Khatik, 2019). The chemical structures of all these deriva-
tives along with their IC50 values are reported in Table 14.
Aza-­ Michael reaction has been used for the synthesis of From the above six derivatives, it was found that com-
1,3-­diaryl-­3-­(arylamino) propan-­1-­one derivatives so as to get pound (e) showed more potency, compound (a) was the
KAUR et al.   
| 15

TABLE 12 Various benzofuran hydrazone derivatives with IC50 TABLE 13 Benzotriazole derivatives and their IC50 value
values
IC50
A IC50 (μm) (μm) for
R1 R2 R3 α-­amylase
OH 1.644 ± 0.128
Cl Cl 2.99
A

OH 2.320 ± 0.05
Cl

A H H 2.80
OH
F
OH 1.245 ± 0.25
OH
H H 2.98
A
OCH3
OH 1.843 ± 0.19
H H 2.92
A OH
OH NO2

O
CH3 1.350 ± 0.24
H H O 2.41
Cl
A
OH
H H Br 2.76
HO OH 1.629 ± 0.05 O

A
OH

O 1.353 ± 0.232
N+ –
O CH3 CH3 O 2.12
Cl
A

Cl 1.359 ± 0.119
CH3 CH3 O 2.51
A OCH3

Cl 1.488 ± 0.07
Standard acarbose 1.98 μm
A

Acarbose 0.62 ± 0.22

R1 N
N
R3
O R2 N
O
N N A FIGURE 15 Benzotriazole moiety

H 3.2.6 | Pyrrole and pyrrolidine derivatives as

CH3
α-­amylase inhibitors
FIGURE 14 Benzofuran hydrazone derivative
The new derivatives of N-­acetylpyrrolidine have been in-
vestigated with an inhibitory α-­amylase activity, which is
shown in Table 16. Out of all the derivatives, two of them
second most potent compound, and compound (d) showed were found to show good inhibitory activity as compared to
antioxidant activity (Bashary & Khatik, 2019). acarbose (Sansenya et al., 2020).
Imines are also found to have an important role in me-
dicinal chemistry. Thus, synthesis of different aryl imine
derivatives has been done. Five aryl imine derivatives 3.2.7 | Pyrole and pyrazolone heterocyclic
have been reported to have more α-­amylase inhibitory ac- compounds as alpha-­amylase inhibitors
tivity (Aispuro-­Pérez et al., 2020) whose chemical struc-
tures are mentioned in Table 15 and Figure 16 with their A report has been submitted about azoles, which were sub-
IC50 values. stituted with sulphanyl groups to produce novel compounds
16
|    KAUR et al.

T A B L E 1 4 Synthesis of various 1, 3-­diaryl-­3-­(arylamino) TABLE 15 Different aryl imine derivatives with their IC50 values
propan-­1-­one derivatives
S. No R′ R″ IC50 (µm)
IC50
1. 2-­C5H4N 4-­NO2 0.125
S. No Compound (µm)
2. 2-­CH3C6H4 4-­NO2 0.186
1. 0.085
3. 3-­CH3C6H4 4-­NO2 0.156
4. 4-­CH3C6H4 4-­NO2 0.217
O NH
5. C6H11 4-­NO2 0.151
Acarbose –­ –­ 0.095

(a)
2. CH3 0.716
R''
N
O NH R'
FIGURE 16 Aryl imine moiety

(b) TABLE 16 Derivatives of N-­acetylpyrrolidine with their IC50

values
3. 0.536
Compound IC50 (µm)
O NH
3,210 ± 0.65

OCH3 N
O
(c)
4. OCH3 0.757

O NH

N-(benzyl)-2-acetylpyrrolidine
2,720 ± 0.09
(d)
5. Cl 0.0689 N

O
O S O
O NH

(e)
6. Br 0.126

N-(tosyl)-2-acetylpyrrolidine
O NH

with excellent inhibitory activity towards alpha-­

amylase.
Four compounds are reported with their characteristics in
(f) Table 17 (Maksimov et al., 2016).
KAUR et al.   
| 17

A rationally prepared set of seven different derivatives extract 200 mg/kg = AM200; and A. megalacantha extract
containing pyrazole moiety on thiazoline-­4-­one scaffold was 400 mg/kg = AM400.
reported recently. Spectroscopic techniques were used for
determining the structure and its properties. An inhibition
percentage was noted at 50, 100 and 200 μg/ml of concentra- 3.2.9 | Oxadiazole derivatives as α-­
tion, and acarbose was preferred as standard for comparison amylase inhibitors
(Kumar et al., 2017). The chemical structures of pyrazole-­
substituted thiazoline-­4-­one derivatives and their percentage Conditions for synthesis of 1, 3, 4–­oxadiazole derivatives
inhibition are shown in Table 18 and Figure 17, respectively. were designed to examine α-­amylase inhibitory activity. X-­
To combat adverse effects, Yousuf et al worked on syn- ray studies were included to investigate structural informa-
thesizing 18 pyrazolone compounds (IC50 ranges between tion, and Gaussian 09 software was used for computational
1.61 and 2.38 μm) substituted with arylidene and aryl rings. calculations. The compounds were formed with a good
Their basic chemical nucleus is shown in Figure 18, and percentage yield of 70%–­80%. All the three compounds
these pyrazolone compounds are reported as type of mixed were investigated, and 2-­ (2-­
(trifluoromethyl) benzylthio)-­
inhibitors on the basis of their kinetic studies (Yousuf 5-­(4-­methoxyphenyl)-­1,3,4-­oxadiazole with IC50 value of
et al., 2018). (0.237 ± 0.23 µm) was found to have a better α-­amylase in-
Duhan et al have reported a series of thiazole compounds hibitory activity than others (Hamdani et al., 2020). Its chem-
clubbed with pyrazole ring for their inhibitory activity to- ical structure is also shown in Figure 22.
wards alpha-­amylase enzyme. A QSAR model was also pre-
pared using the Monte Carlo method of optimization and an
estimation regarding % inhibition carried out. The substi- 3.2.10 | Miscellaneous active constituents as
tution on basic nucleus for two most potent compounds is α-­amylase inhibitors
shown in Table 19 and Figure 19 with the best activity at
50 μg/ml (Duhan et al., 2019). In 2001, Andel et al. introduced α-­amylase inhibitors and a
Taha et al after searching many pharmacological activi- quite new group of oral antidiabetic drugs; that is, thiazo-
ties of pyrrole and thiophene heterocycles decided to work lidinediones were introduced simultaneously. Blood sugar
on producing new series with the combination of them so levels were monitored for 24 hr with the help of a subcu-
as to get some novel compounds with better alpha-­amylase taneous glucose sensor (Andel, 2001). Synthesis of phlo-
inhibitory activity. The best potent activity was seen in roglucinol, a derivative and two other known compounds
the underlying compound shown in Figure 20 (Eldebss that were isolated from Eisenia bicyclis (brown alga), was
et al., 2019). carried out. Inhibitory activity was observed on α-­amylase
and glycation (Okada et al., 2004). Further, it was found that
sprouted or bioprocessed soya beans were found to have
3.2.8 | Active constitutes of Aloe extract as α-­ improved inhibitory action and bioprocessing or long-­term
amylase inhibitors sprouting seemed to potentiate glucosidase inhibitory activ-
ity (P. McCue et al., 2005). Later, Itoh et al. have reported
Recently, Tekulu et al. have reported the antidiabetic activity aqueous extract of Vigna angularis (adzuki beans) as poten-
of two polar isolates (AM1 and AG1), each of Aloe monticola tial inhibitor of α-­amylase activity (Itoh et al., 2004). This
Reynolds and Aloe megalacantha baker extracts of leaf latex activity may be associated with L/D-­chiro-­inositol, ononitol,
via in vitro inhibition of α-­amylase at the IC50 value of 37.83 galactosylononitol, pinitol, sequoyitol, L/D-­bornesitol, and
and 56.95 μg/ml, respectively (Tekulu et al., 2019). Active L-­quebrachitol, which are present in germinating seeds of
constituents, 7-­O-­methyl-­6′-­O-­coumaroylaloesin and aloesin beans including Vigna angularis (Peterbauer et al., 2003).
of A. megalacantha baker leaf latex, have been shown to have Some of the plant extracts were investigated where a close
antidiabetic activity (Hiruy et al., 2019). The graphical presen- analysis of 126 extracts of 17 medicinal plants was taken
tation of in vitro study for A. monticola Reynold and A. meg- under consideration for the inhibition of porcine pancreatic
alacantha baker is shown in Figure 21 with their IC50 values. amylase. Only three of the isopropyl extracts were found to
In the same year, Hammeso et al. (2019) have also men- have more than 50% inhibitory concentration activity, which
tioned antihyperlipidaemic and antidiabetic effects of A. me- is reported in Table 21 (Milella et al., 2016).
galacantha extract obtained from leaf latex. Table 20 shows Silver nanoparticles were prepared from aqueous leaf ex-
the antidiabetic effects on streptozotocin (STZ)-­induced dia- tract of Lonicera japonica and silver nitrate. These nanopar-
betic model (Hammeso et al. 2019). ticles were found to be effective reversible non-­competitive
Diabetic control = DC; glibenclamide 5 mg/kg = GL5; A. inhibitors of α-­amylase and α-­glucosidase at IC50 values of
megalacantha extract 100 mg/kg = AM100; A. megalacantha 54.56 and 37.86 µg/ml, respectively (Lu et al., 2017).
18
|    KAUR et al.

T A B L E 1 7 Substituted azoles along

S. No. Compound structure Characteristics
with their characteristics
1. N Competitive inhibitor
NH
S
S
HN
N

2. N Competitive inhibitor
NH
S
S
HN .2HCL
N

3. N Competitive inhibitor
O
S
S
O
N

4. HN Non-­competitive
N inhibitor
S

T A B L E 1 8 Newly prepared alpha-­amylase inhibitors with

percentage inhibition

Compound % inhibition
S. No name R (at 50 μm)
1. 5a –­OCH3 75.94
FIGURE 18 Arylidene-­and aryl-­substituted pyrazolone moiety
2. 5b –­CH3 79.24
3. 5c –­H 48.08
TABLE 19 Derivatives of thiazole attached to pyrrole ring
4. 5d –­Cl 50.94
5. 5e –­F 77.12 S. No Compound R1 R2
6. 5f –­Br 52.12 1. 1 (4g) -­CH3 -­Br
7. 5g –­NO2 65.09 2. 2 (4h) -­CH3 -­Cl
8. Acarbose –­ 48.58

4 | CONCLUSION

R There have been numerous studies published on the inhibi-

tory activity of various compounds and their derivatives
on the alpha-­amylase enzyme in the treatment of diabetes.
N Amylase is an enzyme produced by acinar cells that breaks
Ph N down polysaccharide molecules into glucose in our bodies,
raising blood glucose levels and causing hyperglycaemia.
This is the reason that α-­amylase is a prominent therapeu-
tic target for the treatment and maintenance of postprandial
S O rise in blood glucose levels. Inhibition of α-­amylase is linked
to the use of drugs such as acarbose, miglitol and voglib-
NH
N ose in the treatment of diabetes. Different derivatives of drug
molecules such as benzofuran hydrazone, indole hydrazone,
FIGURE 17 Thiazoline-­4-­one substituted with pyrazole ring spiroindolone, benzotriazoles, 1,3-­diaryl-­3-­(arylamino)
KAUR et al.   
| 19

R2 T A B L E 2 0 Leaf latex extract of A. megalacantha and effect in

STZ-­induced diabetic mice

Blood glucose Blood glucose Blood glucose

R1 level (mg/dl) level (mg/dl) level (mg/dl)
N
Group Day 0 Day 7 Day 14
HN
S DC 264.33 266.83 268.83
N
GL5 256.33 219.50 191.67
AM100 265.33 241.50 203.83
N
N AM200 262.17 239.50 204.00
Ph AM400 262.17 241.17 202.00
Note: Diabetic control = DC; glibenclamide 5 mg/kg = GL5; A. megalacantha
FIGURE 19 Thiazole moiety attached with heterocyclic ring extract 100 mg/kg = AM100; A. megalacantha extract 200 mg/kg = AM200; A.
megalacantha extract 400 mg/kg = AM400.

O
SO2Ph
F3C
O
N N
S
O
N
Ph CH3
N O

H3C F I G U R E 2 2 2-­(2-­(trifluoromethyl) benzylthio)-­5-­(4-­

methoxyphenyl)-­1, 3, 4-­oxadiazole

IC50= 63.66 μM
T A B L E 2 1 Name of plants and their extracts expressing ≥50%
FIGURE 20 Pyrole and thiophene heterocyclic derivative PPA inhibition

Plant name Extracted compound

O. tenuiflorum Methyleugenol
Camphene
5-­Ethyl-­2-­heptanol
M. alba 9,12-­Octadecadienoic acid
Hexadecanoic acid
Napthelene
L. usitatisumum Cyclopentaneundecanoic acid
2-­Cyclopentene-­1-­undecanoic acid

of these drug derivatives have been determined by their IC50

values, drug–­receptor interactions, biological activities, and
a variety of other factors. Methodological and modern ap-
F I G U R E 2 1 In vitro study and IC50 of A. monticola Reynold proaches, such as computer-­aided drug designing and discov-
and A. megalacantha baker ery, may be very helpful in future to overcome the challenges
in development of drugs for the treatment of diabetes caused
by α-­amylase inhibition.
propan-­1-­one, oxadiazole and flavonoids, and others have
been developed by researchers. Flavonoids have a wide range ACKNOWLEDGEMENT
of interactions that inhibit alpha-­amylase. These pyrrole and The authors are thankful to the Head of the Pharmacy of
pyrazolone derivatives have piqued researchers' interest in re- Lovely Professional University, Jalandhar, Punjab, for pro-
cent years as potential new moieties. The therapeutic effects viding facilities to carry out the work.
20
|    KAUR et al.

CONFLICT OF INTEREST Eck, P. (2013). Recombinant DNA Technologies in Food Biochemistry of

All authors have no conflict of interests regarding the publi- Foods (pp. 503–­556). Elsevier.
Eldebss, T. M. A., Farag, A. M., Abdalla, M. M., & Khedr, A. A. (2019).
cation of this paper.
Synthesis of some new pyrazolone-­based heterocycles containing
sulphone moiety acting as α-­glucosidase and α-­amylase inhibitors.
R E F E R E NC E S Journal of Heterocyclic Chemistry, 56(3), 765–­ 780. https://doi.
Agarwal, P. (2016). Alpha-­amylase inhibition can treat diabetes melli- org/10.1002/jhet.3433
tus. Research and Reviews Journal of Medical and Health Sciences, Eleazu, C., Charles, A., Eleazu, K., & Achi, N. (2018). Free fatty acid
5(4), 1–­8. receptor 1 as a novel therapeutic target for type 2 diabetes mellitus-­
Agarwal, P., & Gupta, R. (2016). Alpha-­amylase inhibition can treat current status. Chemico-­biological Interactions, 289, 32–­39. https://
diabetes mellitus. Research and Reviews: Journal of Medical Health doi.org/10.1016/j.cbi.2018.04.026
Sciences, 5(4), 1–­8. Fox, G. (2018). Starch in brewing applications starch in food (pp. 633–­
Aispuro-­Pérez, A., López-­Ávalos, J., García-­Páez, F., Montes-­Avila, J., 659). Elsevier.
Picos-­Corrales, L. A., Ochoa-­Terán, A., Bastidas, P., Montaño, S., Geng, P., Bai, G., Shi, Q., Zhang, L., Gao, Z., & Zhang, Q. (2009).
Calderón-­Zamora, L., Osuna-­Martínez, U., & Sarmiento-­Sánchez, Taxonomy of the Streptomyces strain ZG0656 that produces
J. I. (2020). Synthesis and molecular docking studies of imines as acarviostatin α-­ amylase inhibitors and analysis of their ef-
α-­glucosidase and α-­amylase inhibitors. Bioorganic Chemistry, 94, fects on blood glucose levels in mammalian systems. Journal of
103491. https://doi.org/10.1016/j.bioorg.2019.103491 Applied Microbiology, 106(2), 525–­533. https://doi.org/10.1111/
Altowyan, M. S., Barakat, A., Al-­Majid, A. M., & Al-­Ghulikah, H. j.1365-­2672.2008.​04021.x
(2019). Spiroindolone analogues as potential hypoglycemic with Goldstein, B. J. (2001). Protein-­tyrosine phosphatase 1B (PTP1B): A
dual inhibitory activity on α-­amylase and α-­glucosidase. Molecules, novel therapeutic target for type 2 diabetes mellitus, obesity and
24(12), 2342.–­https://doi.org/10.3390/molec​ules2​4122342 related states of insulin resistance. Current Drug Targets-­Immune,
Andel, M. (2001). Diabetology at the threshold of the 21st century. Endocrine & Metabolic Disorders, 1(3), 265–­ 275. https://doi.
Vnitrni Lekarstvi, 47(5), 277–­280. org/10.2174/15680​08013​341163
Baker, D. J., Timmons, J. A., & Greenhaff, P. L. (2005). Glycogen Hamdani, S. S., Khan, B. A., Ahmed, M. N., Hameed, S., Akhter, K.,
phosphorylase inhibition in type 2 diabetes therapy: A systematic Ayub, K., & Mahmood, T. (2020). Synthesis, crystal structures,
evaluation of metabolic and functional effects in rat skeletal mus- computational studies and α-­amylase inhibition of three novel 1,
cle. Diabetes, 54(8), 2453–­ 2459. https://doi.org/10.2337/diabe​ 3, 4-­oxadiazole derivatives. Journal of Molecular Structure, 1200,
tes.54.8.2453 127085. https://doi.org/10.1016/j.molst​ruc.2019.127085
Balan, K., Qing, W., Wang, Y., Liu, X., Palvannan, T., Wang, Y., Ma, Hameed, S., Kanwal, X. X., Seraj, F., Rafique, R., Chigurupati, S.,
F., & Zhang, Y. (2016). Antidiabetic activity of silver nanoparti- Wadood, A., Rehman, A. U., Venugopal, V., Salar, U., Taha, M.,
cles from green synthesis using Lonicera japonica leaf extract. RSC & Khan, K. M. (2019). Synthesis of benzotriazoles derivatives and
Advances, 6(46), 40162–­ 40168. https://doi.org/10.1039/C5RA2​ their dual potential as alpha-­amylase and alpha-­glucosidase inhib-
4391B itors in vitro: Structure-­activity relationship, molecular docking,
Bashary, R., & Khatik, G. L. (2019). Design, and facile synthesis of and kinetic studies. European Journal of Medicinal Chemistry, 183,
1, 3 diaryl-­3-­(arylamino) propan-­1-­one derivatives as the potential 111677. https://doi.org/10.1016/j.ejmech.2019.111677
alpha-­amylase inhibitors and antioxidants. Bioorganic Chemistry, Hammeso, W. W., Emiru, Y. K., Ayalew Getahun, K., & Kahaliw, W.
82, 156–­162. https://doi.org/10.1016/j.bioorg.2018.10.010 (2019). Antidiabetic and antihyperlipidemic activities of the leaf latex
Blair, M. (2016). Diabetes Mellitus review. Urologic Nursing, 36(1). extract of Aloe megalacantha baker (Aloaceae) in streptozotocin-­
https://doi.org/10.7257/1053-­816X.2016.36.1.27 induced diabetic model. Evidence-­ Based Complementary and
Butterworth, P. J., Warren, F. J., & Ellis, P. R. (2011). Human α-­amylase Alternative Medicine. https://doi.org/10.1155/2019/8263786
and starch digestion: An interesting marriage. Starch -­Stärke, 63(7), Hemlata, B., Pornima, G., Tukaram, K., & Pankaj, B. (2019). In vitro
395–­405. https://doi.org/10.1002/star.20100​0150 anti-­amylase activity of some Indian dietary spices. Journal of
Crepaldi, G., Carruba, M., Comaschi, M., Del Prato, S., Frajese, G., Applied Biology & Biotechnology, 7(04), 70–­74.
& Paolisso, G. (2007). Dipeptidyl peptidase 4 (DPP-­ 4) inhibi- Hiruy, M., Bisrat, D., Mazumder, A., & Asres, K. (2019). Two chro-
tors and their role in Type 2 diabetes management. Journal of mones with antimicrobial activity from the leaf latex of Aloe mon-
Endocrinological Investigation, 30(7), 610–­ 614. https://doi. ticola Reynolds. Natural Product Research, 35, 1–­5. https://doi.
org/10.1007/BF033​46357 org/10.1080/14786​419.2019.1614581
Derosa, G., & Maffioli, P. (2012). Peroxisome proliferator-­activated re- Hwang, J. L., & Weiss, R. E. (2014). Steroid-­ induced diabetes: A
ceptor-­γ (PPAR-­γ) agonists on glycemic control, lipid profile and clinical and molecular approach to understanding and treatment.
cardiovascular risk. Current Molecular Pharmacology, 5(2), 272–­ Diabetes/metabolism Research and Reviews, 30(2), 96–­102. https://
281. https://doi.org/10.2174/18744​67211​20502​0272 doi.org/10.1002/dmrr.2486
Des Gachons, C. P., & Breslin, P. A. (2016). Salivary amylase: Digestion Itoh, T., Kita, N., Kurokawa, Y., Kobayashi, M., Horio, F., & Furuichi,
and metabolic syndrome. Current Diabetes Reports, 16(10), 102. Y. (2004). Suppressive effect of a hot water extract of adzuki beans
https://doi.org/10.1007/s1189​2-­016-­0794-­7 (Vigna angularis) on hyperglycemia after sucrose loading in mice
Duhan, M., Singh, R., Devi, M., Sindhu, J., Bhatia, R., Kumar, A., & and diabetic rats. Bioscience, Biotechnology, and Biochemistry,
Kumar, P. (2019). Synthesis, molecular docking and QSAR study of 68(12), 2421–­2426. https://doi.org/10.1271/bbb.68.2421
thiazole clubbed pyrazole hybrid as α-­amylase inhibitor. Journal of Jain, N. K., Boivin, M., Zinsmeister, A. R., Brown, M. L., Malagelada,
Biomolecular Structure and Dynamics, 39(1), 91–­107. https://doi. J. R., & DiMagno, E. P. (1989). Effect of ileal perfusion of car-
org/10.1080/07391​102.2019.1704885 bohydrates and amylase inhibitor on gastrointestinal hormones
KAUR et al.   
| 21

and emptying. Gastroenterology, 96(2 Pt 1), 377–­387. https://doi. Mahmood, N. (2016). A review of α-­amylase inhibitors on weight loss
org/10.1016/0016-­5085(89)91561​-­8 and glycemic control in pathological state such as obesity and diabe-
Kahanovitz, L., Sluss, P. M., & Russell, S. J. (2017). Type 1 diabe- tes. Comparative Clinical Pathology, 25(6), 1253–­1264. https://doi.
tes–­
a clinical perspective. Point of Care, 16(1), 37. https://doi. org/10.1007/s0058​0-­014-­1967-­x
org/10.1097/POC.00000​00000​000125 Maksimov, V., Zaynullin, R., Akhmadiev, N., Segura-­Ceniceros, E.
Kantsadi, A. L., Parmenopoulou, V., Bakalov, D. N., Snelgrove, P., Martínez Hernández, J. L., Bikbulatova, E., Akhmetova, V.,
L., Stravodimos, G. A., Chatzileontiadou, D. S. M., Manta, S., Kunakova, R., Ramos, R., & Ilyina, A. (2016). Inhibitory effect of 4,
Panagiotopoulou, A., Hayes, J. M., & Komiotis, D. D. (2015). 4′-­[ethane-­1,2-­diylbis(sulfandiylmethanediyl)]bis(3,5-­dimethyl-­1H-­
Glycogen phosphorylase as a target for type 2 diabetes: Synthetic, pyrazole) and its derivatives on alpha-­amylase activity. Medicinal
biochemical, structural and computational evaluation of novel N-­ Chemistry Research, 25(7), 1384–­ 1389. https://doi.org/10.1007/
acyl-­N´-­(β-­D-­glucopyranosyl) urea inhibitors. Current Topics in s0004​4-­016-­1574-­2
Medicinal Chemistry, 15(23), 2373–­2389. Martinez-­Gonzalez, A. I., Díaz-­Sánchez, Á., de La Rosa, L., Bustos-­Jaimes,
Kaur, R., Dahiya, L., & Kumar, M. (2017). Fructose-­1, 6-­bisphosphatase I., Alvarez-­Parrilla, E. J. S. A. P. A. M., & Spectroscopy, B. (2019).
inhibitors: A new valid approach for management of type 2 diabetes Inhibition of α-­amylase by flavonoids: Structure activity relationship
mellitus. European Journal of Medicinal Chemistry, 141, 473–­505. (SAR). Spectrochimica Acta Part A: Molecular and Biomolecular
https://doi.org/10.1016/j.ejmech.2017.09.029 Spectroscopy, 206, 437–­447. https://doi.org/10.1016/j.saa.2018.08.057
Kazeem, M. I., Ogunbiyi, J. V., & Ashafa, A. (2013). In vitro studies Matsui, T., Ueda, T., Oki, T., Sugita, K., Terahara, N., & Matsumoto,
on the inhibition of α-­amylase and α-­glucosidase by leaf extracts K. (2001). α-­Glucosidase Inhibitory Action of Natural Acylated
of Picralima nitida (Stapf). Tropical Journal of Pharmaceutical Anthocyanins. 1. Survey of Natural Pigments with Potent Inhibitory
Research, 12(5), 719–­725. Activity. Journal of Agricultural and Food Chemistry, 49(4), 1948–­
Kharroubi, A. T., & Darwish, H. M. (2015). Diabetes mellitus: The ep- 1951. https://doi.org/10.1021/jf001​251u
idemic of the century. World Journal of Diabetes, 6(6), 850–­867. McCue, P., Kwon, Y. I., & Shetty, K. (2005). Anti-­diabetic and anti-­
https://doi.org/10.4239/wjd.v6.i6.850 hypertensive potential of sprouted and solid-­state bioprocessed soy-
Kianbakht, S., Abasi, B., & Dabaghian, F. H. (2013). Anti-­hyperglycemic bean. Asia Pacific Journal of Clinical Nutrition, 14(2), 145–­152.
effect of Vaccinium arctostaphylos in type 2 diabetic patients: A McCue, P. P., & Shetty, K. (2004). Inhibitory effects of rosmarinic acid
randomized controlled trial. Forsch Komplementmed, 20(1), 17–­22. extracts on porcine pancreatic amylase in vitro. Asia Pacific Journal
https://doi.org/10.1159/00034​6607 of Clinical Nutrition, 13(1), 101–­106.
Kim, J. S., Kwon, C. S., & Son, K. H. (2000). Inhibition of alpha-­ Merger, S. R., Kerner, W., Stadler, M., Zeyfang, A., Jehle, P., Müller-­
glucosidase and amylase by luteolin, a flavonoid. Bioscience, Korbsch, M., & Holl, R. W. (2016). Prevalence and comorbidities
Biotechnology, and Biochemistry, 64(11), 2458–­2461. https://doi. of double diabetes. Diabetes Research and Clinical Practice, 119,
org/10.1271/bbb.64.2458 48–­56. https://doi.org/10.1016/j.diabr​es.2016.06.003
Kitamoto, K. (2002). Molecular Biology of the Koji Molds. In A. I. Milella, L., Milazzo, S., De Leo, M., Vera Saltos, M. B., Faraone, I.,
Laskin, J. W. Bennett, & G. M. Gadd (Eds.), Advances in applied Tuccinardi, T., & Braca, A. (2016). alpha-­Glucosidase and alpha-­
microbiology, Vol. 51 (pp. 129–­153). Academic Press. Amylase Inhibitors from Arcytophyllum thymifolium. Journal of
Kneuer, C., Ehrhardt, C., Radomski, M. W., & Bakowsky, U. (2006). Natural Products, 79(8), 2104–­ 2112. https://doi.org/10.1021/acs.
Selectins–­ potential pharmacological targets? Drug Discovery jnatp​rod.6b00484
Today, 11(21–­22), 1034–­1040. https://doi.org/10.1016/j. Mojsov, K. (2016). Aspergillus enzymes for food industries. New and
drudis.2006.09.004 future developments in microbial biotechnology and bioengineering
Kumar, P., Duhan, M., Kadyan, K., Sindhu, J., Kumar, S., & Sharma, (pp. 215–­222). Elsevier.
H. (2017). Synthesis of novel inhibitors of α-­amylase based on the Morris, C., Fichtel, S. L., & Taylor, A. J. (2011). Impact of calcium
thiazolidine-­4-­one skeleton containing a pyrazole moiety and their on salivary α-­amylase activity, starch paste apparent viscosity, and
configurational studies. MedChemComm, 8(7), 1468–­1476. https:// thickness perception. Chemosensory Perception, 4(3), 116. https://
doi.org/10.1039/C7MD0​0080D doi.org/10.1007/s1207​8-­011-­9091-­7
Lee, Y. H., Wang, M.-­Y., Yu, X.-­X., & Unger, R. H. (2016). Glucagon is Noreen, T., Taha, M., Imran, S., Chigurupati, S., Rahim, F., Selvaraj,
the key factor in the development of diabetes. Diabetologia, 59(7), M., Ismail, N. H., Mohammad, J. I., Ullah, H., javid, M. T., Nawaz,
1372–­1375. https://doi.org/10.1007/s0012​5-­016-­3965-­9 F., Irshad, M., & Ali, M. (2017). Synthesis of alpha amylase inhibi-
Lemelman, M. B., Letourneau, L., & Greeley, S. A. W. (2018). Neonatal tors based on privileged indole scaffold. Bioorganic Chemistry, 72,
Diabetes Mellitus: an update on diagnosis and management. 248–­255. https://doi.org/10.1016/j.bioorg.2017.04.010
Clinics in Perinatology, 45(1), 41–­ 59. https://doi.org/10.1016/j. Oboh, G., Ogunsuyi, O. B., Ogunbadejo, M. D., & Adefegha, S. A.
clp.2017.10.006 (2016). Influence of gallic acid on α-­amylase and α-­glucosidase in-
Lo Piparo, E., Scheib, H., Frei, N., Williamson, G., Grigorov, M., & hibitory properties of acarbose. Journal of Food and Drug Analysis,
Chou, C. J. (2008). Flavonoids for controlling starch digestion: 24(3), 627–­634. https://doi.org/10.1016/j.jfda.2016.03.003
Structural requirements for inhibiting human alpha-­ amylase. Okada, Y., Ishimaru, A., Suzuki, R., & Okuyama, T. (2004). A new phlo-
Journal of Medicinal Chemistry, 51(12), 3555–­3561. https://doi. roglucinol derivative from the brown alga Eisenia bicyclis: Potential
org/10.1021/jm800​115x for the effective treatment of diabetic complications. Journal of
Lu, Y., Zhou, W., Feng, Y., Li, Y., Liu, K., Liu, L., & Wu, X. (2017). Natural Products, 67(1), 103–­ 105. https://doi.org/10.1021/np030​
Acteoside and acyl-­ migrated acteoside, compounds in Chinese 323j
kudingcha tea, inhibit alpha-­amylase in vitro. Journal of Medicinal Sudha, P., Zinjarde, S. S., Bhargava, S. Y., & Kumar, A. R. (2011).
Food, 20(6), 577–­585. https://doi.org/10.1089/jmf.2016.3910 Potent alpha-­ amylase inhibitory activity of Indian Ayurvedic
22
|    KAUR et al.

medicinal plants. BMC Complementary and Alternative Medicine, as novel alpha-­amylase inhibitor. Bioorganic Chemistry, 75, 78–­85.
11, 5. https://doi.org/10.1186/1472-­6882-­11-­5 https://doi.org/10.1016/j.bioorg.2017.09.002
Peterbauer, T., Brereton, I., & Richter, A. (2003). Identification of a Tekulu, G. H., Araya, E. M., & Mengesha, H. G. (2019). In vitro alpha-­
digalactosyl ononitol from seeds of adzuki bean (Vigna angu- amylase inhibitory effect of TLC isolates of Aloe megalacantha
laris). Carbohydrate Research, 338(19), 2017–­ 2019. https://doi. baker and Aloe monticola Reynolds. BMC Complementary and
org/10.1016/S0008​-­6215(03)00339​-­2 Alternative Medicine, 19(1), 206. https://doi.org/10.1186/s1290​
Pozzilli, P., & Pieralice, S. (2018). Latent autoimmune diabetes in 6-­019-­2622-­5
adults: Current status and new horizons. Endocrinology and Tiwari, N., Thakur, A. K., Kumar, V., Dey, A., & Kumar, V. (2014).
Metabolism (Seoul, Korea), 33(2), 147–­159. https://doi.org/10.3803/ Therapeutic targets for diabetes mellitus: An update. Clinical
EnM.2018.33.2.147 Pharmacology & Biopharmaceutics, 3(1), 1. https://doi.
Proenca, C., Freitas, M., Ribeiro, D., Tomé, S. M., Oliveira, E. F. org/10.4172/2167-­065X.1000117
T., Viegas, M. F., Araújo, A. N., Ramos, M. J., Silva, A. M. S., Toldra, F., & Kim, S.-­ K. (2016). Marine enzymes biotechnology:
Fernandes, P. A., & Fernandes, E. (2019). Evaluation of a flavo- Production and industrial applications, part II-­marine organisms
noid’s library for inhibition of pancreatic α-­ amylase towards a producing enzymes. Academic Press.
structure-­activity relationship. J Enzyme Inhib Med Chem., 34(1), Tomlinson, D. R., Stevens, E. J., & Diemel, L. T. (1994). Aldose reduc-
577–­588. tase inhibitors and their potential for the treatment of diabetic com-
Qin, X., Ren, L., Yang, X., Bai, F., Wang, L., Geng, P., & Shen, Y. plications. Trends in Pharmacological Sciences, 15(8), 293–­297.
(2011). Structures of human pancreatic alpha-­amylase in complex https://doi.org/10.1016/0165-­6147(94)90010​-­8
with acarviostatins: Implications for drug design against type II di- Tucci, S. A., Boyland, E. J., & Halford, J. C. (2010). The role of lipid
abetes. Journal of Structural Biology, 174(1), 196–­202. https://doi. and carbohydrate digestive enzyme inhibitors in the management
org/10.1016/j.jsb.2010.11.020 of obesity: A review of current and emerging therapeutic agents.
Rizos, E. C., & Elisaf, M. S. (2014). Sodium-­glucose co-­transporter 2 Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy,
inhibition in diabetes treatment: Current evidence and future per- 3, 125–­143. https://doi.org/10.2147/dmsott.s7005
spectives. Current Pharmaceutical Design, 20(22), 3647–­3656. Urbanová, J., Brunerová, L., & Brož, J. (2018). Hidden MODY—­
Rossi, E. J., Sim, L., Kuntz, D. A., Hahn, D., Johnston, B. D., Looking for a Needle in a Haystack. Frontiers in Endocrinology,
Ghavami, A., Szczepina, M. G., Kumar, N. S., Sterchi, E. E., 9(355). https://doi.org/10.3389/fendo.2018.00355
Nichols, B. L., Pinto, B. M., & Rose, D. R. (2006). Inhibition van de Laar, F. A. (2008). Alpha-­glucosidase inhibitors in the early treat-
of recombinant human maltase glucoamylase by salacinol and ment of type 2 diabetes. Vascular Health and Risk Management,
derivatives. FEBS Journal, 273(12), 2673–­ 2683. https://doi. 4(6), 1189.
org/10.1111/j.1742-­4658.2006.05283.x Vantyghem, M. C., & Press, M. (2006). Management strategies for brit-
Saini, R., Saini, H. S., & Dahiya, A. (2017). Amylases: Characteristics tle diabetes. Annales D'endocrinologie, 67(4), 287–­294. https://doi.
and industrial applications. J. Pharmacogn. Phytochem, 6(4), org/10.1016/S0003​-­4266(06)72600​-­2
1865–­1871. Wall, S., Chow, J., Huang, J., Jiang, M., Yang, A., & Jones, K. (2015).
Sales, P. M., Souza, P. M., Simeoni, L. A., & Silveira, D. (2012). alpha-­ Phosphoenolpyruvate Carboxykinase: Possible Therapeutic Targets
Amylase inhibitors: A review of raw material and isolated com- for Insulin Resistant Type-­II Diabetes. The FASEB Journal, 29(1_
pounds from plant source. Journal of Pharmacy and Pharmaceutical supplement), LB67. https://doi.org/10.1096/fasebj.29.1_suppl​
Sciences, 15(1), 141–­183. https://doi.org/10.18433/​j35s3k ement.lb67
Samrot, A., & Vijay, A. (2008). A-­Amylase activity of wild and mutant Yousuf, S., Khan, K. M., Salar, U., Chigurupati, S., Muhammad, M. T.,
strains of Bacillus sp. The Internet Journal of Microbiology, 6(2), Wadood, A., Aldubayan, M., Vijayan, V., Riaz, M., & Perveen, S.
1–­4. (2018). 2ʹ-­Aryl and 4ʹ-­arylidene substituted pyrazolones: As poten-
Sansenya, S., Winyakul, C., Nanok, K., & Phutdhawong, W. S. (2020). tial α-­amylase inhibitors. European Journal of Medicinal Chemistry,
Synthesis and inhibitory activity of N-­ acetylpyrrolidine deriva- 159, 47–­58. https://doi.org/10.1016/j.ejmech.2018.09.052
tives on α-­glucosidase and α-­amylase. Research in Pharmaceutical Zhang, R., Yip, V. L., & Withers, S. G. (2010). Mechanisms of en-
Sciences, 15(1), 14–­25. https://doi.org/10.4103/1735-­5362.278711 zymatic glycosyl transfer. Comprehensive Natural Products II:
Sebastiani, G., Ceccarelli, E., Castagna, M. G., & Dotta, F. (2018). G-­ Chemistry and Biology, 8. https://doi.org/10.1016/B978-­00804​
protein-­coupled receptors (GPCRs) in the treatment of diabetes: 5382-­8.00167​-­2
Current view and future perspectives. Best Practice & Research
Clinical Endocrinology & Metabolism, 32(2), 201–­213. https://doi.
org/10.1016/j.beem.2018.02.005 How to cite this article: Kaur, N., Kumar, V., Nayak,
Shankaraia, P., & Reddy, Y. N. (2011). α-­amylase expressions in Indian S. K., Wadhwa, P., Kaur, P., & Sahu, S. K. (2021).
Type-­2 diabetic patients. Journal of Medical Sciences (Faisalabad), Alpha-­amylase as molecular target for treatment of
11, 280–­284. https://doi.org/10.3923/jms.2011.280.284
diabetes mellitus: A comprehensive review. Chemical
Smith, E., Dion, G., & Morton, M. D. (2010). The mouth, salivary
glands and oesophagus Margaret in The Digestive System. , 2nd ed.
Biology & Drug Design, 00, 1–­22. https://doi.
Taha, M., Shah, S. A. A., Imran, S., Afifi, M., Chigurupati, S., Selvaraj, org/10.1111/cbdd.13909
M., Rahim, F., Ullah, H., Zaman, K., & Vijayabalan, S. (2017).
Synthesis and in vitro study of benzofuran hydrazone derivatives