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Inhibition of Insulin Degrading Enzyme to Control Diabetes Mellitus and its

Applications on some Other Chronic Disease: a Critical Review

Article  in  Pharmaceutical Research · April 2022

DOI: 10.1007/s11095-022-03237-7

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Pharmaceutical Research
https://doi.org/10.1007/s11095-022-03237-7

EXPERT REVIEW

Inhibition of Insulin Degrading Enzyme to Control Diabetes Mellitus

and its Applications on some Other Chronic Disease: a Critical Review
Md. Shofiul Azam1   · Md. Wahiduzzaman2 · Md. Reyad‑ul‑Ferdous3 · Md. Nahidul Islam4 · Mukta Roy5

Received: 15 January 2022 / Accepted: 14 March 2022

© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022

Abstract
Purpose  This review aims to provide a precise perceptive of the insulin-degrading enzyme (IDE) and its relationship to type
2 diabetes (T2D), Alzheimer’s disease (AD), obesity, and cardiovascular diseases. The purpose of the current study was to
provide clear idea of treating prevalent diseases such as T2D, and AD by molecular pharmacological therapeutics rather
than conventional medicinal therapy.
Methods  To achieve the aims, molecular docking was performed using several softwares such as LIGPLOT+, Python, and
Protein-Ligand Interaction Profiler with corresponding tools.
Results  The IDE is a large zinc-metalloprotease that breakdown numerous pathophysiologically important extracellular
substrates, comprising amyloid β-protein (Aβ) and insulin. Recent studies demonstrated that dysregulation of IDE leads
to develop AD and T2D. Specifically, IDE regulates circulating insulin in a variety of organs via a degradation-dependent
clearance mechanism. IDE is unique because it was subjected to allosteric activation and mediated via an oligomer structure.
Conclusion  In this review, we summarised the factors that modulate insulin reformation by IDE and interaction of IDE and
some recent reports on IDE inhibitors against AD and T2D. We also highlighted the latest signs of progress of the function
of IDE and challenges in advancing IDE- targetted therapies against T2D and AD.

KEY WORDS  amyloid-peptide · IDE · insulin · T2D and Alzheimer’s disease

Introduction
Insulin protease and insulinase is an insulin-degrading
enzyme (IDE), a zinc metalloprotease that attached and
changed several types of bioactive peptides by correspond-
ing their sequences and structures, inhibiting peptide aggre-
gation formation to the numerous subcellular compartments
(1). IDE quantified to the 110 kDa zinc protease of the M16
family, which is involved in the degradation and release of
highly preserved insulin (2, 3), amyloid-β (Aβ) peptides
(4), Insulin-like growth factor II (IGF-II) (5), glucagon (6),
amylin (7) as well as somatostatin (8). It plays an active
role in reducing glucagon, β-amyloid, and chemokine ligand
3, among other peptide substrates, through a high insulin

2
* Md. Shofiul Azam Bio‑Med Big Data Center, CAS Key Laboratory
shofiul@duet.ac.bd of Computational Biology, CAS‑MPG Partner Institute
for Computational Biology, Shanghai Institute of Nutrition
Md. Wahiduzzaman
and Health, University of Chinese Academy of Sciences,
wahid@picb.ac.cn
Chinese Academy of Sciences, Shanghai 200031, China
Md. Reyad‑ul‑Ferdous 3
Department of Endocrinology and Metabolism, Shandong
rockyreyad@sdu.edu.cn
Provincial Hospital affiliated to Shandong University,
Md. Nahidul Islam Shandong University, Jinan 250021, Shandong, China
nahidul.islam@bsmrau.edu.bd 4
Department of Agro‑Processing, Bangabandhu Sheikh
Mukta Roy Mujibur Rahman Agricultural University, Gazipur 1706,
muktaroy-fet@sust.edu Bangladesh
5
1 Department of Food Engineering and Tea Technology,
Department of Chemical and Food Engineering, Dhaka
Shahjalal University of Science and Technology,
University of Engineering & Technology, Gazipur 1707,
Sylhet 3114, Bangladesh
Bangladesh

13
Vol.:(0123456789)

sensitivity. The IDE was present in the cellular membranes,
cytoplasm, and several cell organelles (peroxisomes, mito-
chondria, and endosomes) and collected from the extracel-
lular region (9–11). Also, it is settled in Escherichia coli
from mammals and is expressed in both insulin-sensitive/
non-sensitive cells, intimating a multifunctional protein (9).
IDE was discovered by Arthur Mirsky in 1949. Mirsky
argued that IDE’s inhibitors would be a perfect anti-diabetic
medication, allowing them to limit insulin breakdown. Mir-
sky also discovered that when the liver brought out the
capacity of an IDE inhibitor and administered it to rabbits,
they improved insulin action. Mirsky and others explored to
produce efficacious IDE inhibitors as potential medications
(12, 13).
IDE inhibitor depreciated the hypoglycemia action of
insulin, indicating potential therapeutic against IDE (14).
Despite these attempts, only a few chemicals that selectively
block IDE are now accessible, except IDE substrates such as
insulin (15). The active site of the IDE reveals in unparal-
leled aspect the crystal structure imparted by Noinaj, et al.
(16), which holds the key to carrying through a significant
and all-embracing IDE inhibitor. Insulin activity demon-
strated to be potentiated by IDE inhibitors both in cultured
cells and in vivo. Highly selective IDE inhibitors showed
substantial anti-diabetic benefits attributed to decreased
insulin catabolism (17).
The implications of apolipoprotein E (APOE) risk alleles
on cerebral, lipid, and cholesterol metabolism leading to
CVD-mediated insulin resistance (IR) are highlighted, as
well as the consequences of cardiovascular risk factors
leading to IR and amyloid formation. IR, genetic, and car-
diovascular risk factors all play a role in amyloid aggrega-
tion, which can lead to the late onset of AD and DM (18).
Interleukin-6 (IL-6) is a proinflammatory cytokine that con-
trols differentiation, migration, proliferation, and cell death
and hence plays a key role in the development of insulin
resistance and the pathogenesis of type 2 diabetes mellitus
(T2DM) (19). Impairment of IDE function is associated with
the pathophysiology of both type-2-diabetes (T2D) and Alz-
heimer’s disease (AD), according to the atlas database from
human and animal modeling studies as well as molecular
epigenetics (20–22). The IDE is a prototypical member of
the zinc-metal prototype of a distinct superfamily that has
various characteristics that differentiate it from the ordinary
metallic prototypes, having a zinc-binding motif (HxxEH)
(inverted) (23) and an unusual tertiary structure (24).
The subcellular localization of IDE is another distinctive
feature: in the cytosol, most of the massive IDE is found,
with minor aggregates found in the mitochondria, peroxi-
somes, and endosomes. The tiny amount of IDE ranges
between 3% to 10% transported to the extracellular space,
where it communicates with recognized IDE substrates like
insulin and Aβ (25). This study focuses on the molecular
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mechanisms of insulin-degrading enzymes and their roles in
pathogenesis and discusses the potential therapeutic options
based on IDE inhibition in human diseases.
Role of Insulin‑Degrading Enzyme (IDE)
Working Mechanism of Insulin‑Degrading Enzyme
(IDE)
In the enzymatic model that refers to 130/159 atoms, the
IDE performs its catalytic action on two peptides of distinct
lengths using zinc-binding proteases, mimicking a part of
the B chain of insulin, using concentration functional theo-
retical methods and hybrid exchange at the gas stage and
interactions in protein environments effective is B3LYP (26).
According to the authors ‘crystal structures, IDE matches
a clamshell, besides two bowl-shaped sections joined using
an amenable hinge (Fig. 1). The protease can transit between
open and closed states with this arrangement. The elongated
hydrogen bonding on both parts of IDE provides a “latch”
that tends to remain the peptide closed (Fig. 1A), according
to Shen, et al. (28). Researchers were able to boost the effi-
cacy of peptides in cleaving a test substrate by up to 40-fold
by addressing the mutations to the enzyme that disrupts the
hydrogen-bond make fast (Fig. 1B). The breakdown of insu-
lin and amyloid protein was efficient (13).
So, what’s the mechanism behind the mutant IDE’s pro-
nounced enzyme activation? Consider a basic double stair
model of the enzyme response to understand that. At first,
an enzyme-substrate complex developed by combining
enzymes and substrates in the reverse process. Then, the
substrate is catalytically cleaved while the reaction products
are simultaneously released. Mutations that boost the pro-
tease’s open state, approve binding substrate for increasing
the reaction’s productivity by speeding up the creation of the
enzyme-substrate complex (29).
Source and Nomenclature of IDE
Although well known as IDE insulin, it is a highly preserved
M16A ­Zn2+ metalloprotein developed in the tissues of bacte-
ria, fungi, plants, animals, and humans brain, liver, and mus-
cles that were first selected by its capacity to rapidly break
down insulin (30). IDE is unique becasue it has a strong
affinity for substrates with a wide range of sequences and
structures. The 200 remnants of the N-terminal also con-
tain some inverted zinc-metalloprotease core motifs (e.g.,
HXXEH). This is why IDE is classified as a member of
the “inverzincins” family. It’s a thiol zinc-metalloendopepti-
dase with a molecular weight of 110 kDa that’s found in
the cytosol, peroxisomes, endosomes, and cell surface (31).
Insulin, amylin, and β-amyloid protein are among the tiny
Pharmaceutical Research

Fig. 1  Insulin-degrading enzyme (IDE) activation pathway. (a) Insulin-degrading enzyme (IDE) is an enzyme that breaks down compounds
linked to Alzheimer’s disease and diabetes. The crystal form of IDE showed a, ‘latch’ mechanism (Red) that holds the enzyme in a closed state,
delaying the entry of the substrate or exit of the cleavage products. (b) Mutations (Blue) that disrupt the latch promote the open conformation of
the enzyme. All of the mutations are more willing to receive the substrates and discharge consequences the typically appearing the IDE, which is
more active (27).

proteins that IDE catalyzes the breakdown of insulin (32).
IDE homologs play the most vital role in the selection of bud
sites and mating in budding yeast (33). The IDE features a
one-of-a-kind regulation mechanism. It has an inferior for-
mulation and sphere organization with yeast mitochondria
refining proteinase (17).
Prospect of IDE Degradation on Insulin
The ability of IDE to damage the A and B chains of the
hormone insulin, primarily at the locations Leu13-Tyr14
and Tyr14-Gln15 (A chain) and Ser9-His10, His10-Leu11,
Glu13-Ala14, Tyr15-Leu16, and Phe25-Tyr26 (B chain), (B
chain was first discovered). This result revealed that IDE
and pitrilysin, a bacterial protease, have a high degree of
similarity and a likely similar proteolytic process. In terms
of primary sequence, the M16A subfamily enzymes are very
similar. About 30% of identity exists between human IDE
and pitrilysin. This resemblance can be found during the
whole of the length of selected proteins; however, it is com-
monly more prominent in the N-terminal portion (34).
Some researchers looked at mice that didn’t have any
activity in the insulysin gene. They discovered that the
insulysin enzyme destroys amyloid peptides as well as insu-
lin. Moreover, even a little reduction in insulysin activity
raised amyloid peptide levels in the brain (35). The involve-
ment of IDE in Alzheimer’s disease is now well understood;
nevertheless, the biological roles of IDE and many other
M16A enzymes are still poorly understood. Because IDE is
found in the cytosol, peroxisomes, endosomes, proteasome
complexes, and the surface of cerebrovascular endothelial
cells, its roles may be reversed (36).
The Substrate Recognition and Catalytic Mechanism
of IDE
All the structural solutions of IDE in complex with the insu-
lin B-chain, glucagon, and amylin shed light on how IDE
interacts with its substrate and how it cleaves. When bound
to IDE, all three substrates have a total of 70 residues and
fit entirely within the crypt, transforming from a -helix to a
-strand shape (28). The substrate can engage non-covalently
with two IDE-N areas: the catalytic site and the postulated
exosite. The N-terminus of the substratum is attached to the
exosite of the enzyme by engaging with a sheet of IDE-N
once it has been trapped (28). The exosite is a highly con-
served area of IDE that may have a regulatory role in sub-
strate selectivity and catalysis (28, 37). This connection acts
in the way that a molecular chain, allowing the C-terminal
end of the substrate to be properly positioned at the catalytic
region where cleavage occurs (38).
The presence of disulfide bonds is a distinguishing feature
of some IDE substrates (Fig. 2). Insulin, for example, has
three disulfide bonds. The cleavage of insulin by IDE, sur-
prisingly, does not necessitate the decrease and disintegra-
tion of these disulfide bonds. Several other IDE substrates
(e.g., amylin, insulin growth factor I and II) have disulfide
linkages, which could affect how they interact with the cata-
lytic chamber of the enzyme. However, disulfide linkages
are not present in all of IDE’s substrates (e.g., glucagon,

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Fig. 2  Human native and

recombinant IDEs are depicted
schematically. The proteom-
ics sequence inferred from the
human IDE cDNA sequence is
referred to as amino acid posi-
tions (GenBank Accession No.
M21188). Where amino acid
sequences that are found in both
natural and recombinant IDEs
are represented with hatched
boxes. The sequence between
the first putative initiation
methionine (Met 1) and IDE’s
likely the start codon which is
denoted by the black box (Met
42). The tag sequences of the
recombinant IDEs are repre-
sented by the shadow color and
open boxes.

amyloid-β). It’s unclear how the presence of disulfide bonds
affects substrate interaction in the crypt, and therefore more
research is needed (39).
IDE Crystal Structure
The active site of zinc-binding is kept within a proteolytic
cavity produced by defined N- terminal and C-terminal
units linked with a loop including 28 amino acid residues,
according to studies aimed at determining the crystal struc-
ture of IDE. The catalytic chamber is large enough to bind
and destroy insulin-like amyloid (A) layers, which contain
amino acid residues 39–43 and respectively (39).
In these circumstances, the active site anchors the N-ter-
minus of peptides with an exosite that is about 30 degrees
out of the way from the catalytic core. This allows the sub-
strates to be properly located, allowing cleavage at the cata-
lytic site. However, because smaller peptides cannot bind the
exosite and the catalytic core simultaneously, the anchoring
and degrading mechanism for substrates less than around
12 amino acids are unknown. In any case, a researcher has
found that the percentages of catalysis of IDE are higher
for small substrates (> 2000 ­s−1) than for lengthy substrates
likely insulin (about 0.56 min −1) (39).
Configuration of IDE
There are two possible configurations for the IDE. The
first is an open conformation, which lets substrates and
products enter and depart. The other is more difficult to
reach the active site in a closed structure because it is
stored in chambers formed by the two concave domains.
The closed-to-open transfer of catalytic conversion cav-
ity facilitates entry into the middle layer. A significant
boost (approximately 40-fold) in the competent catalytic
of the enzyme is disclosed if mutations inhibit the closed
conformation (37).
This finding showed that altering IDE’s structural prefer-
ence to the open state could be a potential Alzheimer’s treat-
ment. This should speed up the breakdown of Aβ, preventing
aggregation and neuron death (40).
The Function of IDE as a Protease of Insulin
The ability of the IDE to impair insulin into multiple frag-
ments in vitro, giving the principle and trivial outcomes,
was its first distinguishing feature. The first cleavage
events take place in the center of the insulin A and B
chains, with no necessary particular amino acids, implying
that IDE recognizes substrates in terms of the sequence of
amino acid on the tertiary structure rather than primary
(41). As a competitive inhibitor, the poor hydrolysis sub-
stratum is proinsulin which is sustained at slow rates, but
IDE has a high sympathy for insulin (Km 0.1 M) (42).
Insulin-like growth factor I (IGF-I) and IGF-II are both
IDE substrates, with IGF-II degrading faster than IGF-I.
Despite this, they were operating as IDE inhibitors (42).
The leader peptide of rat prethiolase B (P27 peptide),
found in peroxisomes, is another powerful inhibitor of
IDE’s insulin-degrading activity (43).

13
Pharmaceutical Research
Molecular and Biochemical Characteristics of IDE
A gene on the human chromosome 10 q23–q25 and mouse
chromosome 19 give rise to IDE as a unique polypeptide
with a molecular weight of approximately 110 kDa. In the
Goto–Kakizaki rat model, IDE coding alterations have been
linked to the development of T2D (15). Fakhrai-Rad, et al. (15)
discovered two missense variants in IDE (H18R and A890V)
that reduce the ability of transfected cells to digest insulin by
31%. They found that the two mutations had a synergistic
effect on insulin breakdown that is perplexing conferred, where
H18R is existent in the intentional mitochondrial region of
IDE generated via substitute translation initiation (10). Farris,
et al. (44) discovered that recombinant IDE with the A980V
mutation alone had a lower level of catalytic ability for both
insulin and Aβ degradation, implying that this mutation is par-
ticularly important for the proteolytic function. Intriguingly,
there was no payoff on insulin breakdown in cell lysates from
IDE-transfected COS cells, implying the outcome is contingent
on ligand-receptor hormone internalization (43).
IDE Is Heat Shock Protein
HSR is a genetic response to a variety of environmental and
physiological stresses that cause the encoding of genes to
incorporate molecular chaperone, proteases, and other pro-
teins (mostly from the heat shock gene superfamily) that
incorporate and incorrectly combine proteins needed to pro-
tect against cellular damage. The Hsp100, Hsp90, Hsp70,
Hsp60, Hsp40 (J-domain proteins), with short heat shock
protein (sHSP) lineages are arranged in terms of molecular
structure with a magnitude of category in the heat shock gene
superfamily. The maximum number of small molecules act
namely molecular chaperones, causing structural changes
which are necessary for protein synthesis, folding, transloca-
tion, assembly, and destruction. Most of the HSPs are stabi-
lizing detachable proteins and resell deformed proteins under
stressful conditions. If proteins are irrevocably switched,
molecular chaperones assist in their transfer to the cell’s pro-
teolytic machinery, namely the ubiquitin-proteasome path-
way. We show that IDE behaves like a heat shock protein and
that up-regulated amounts of the enzyme can be seen in CNS
malignancies in vivo. According to the findings, IDE performs
a vital role in neuroblastoma cell proliferation via interacting
with the ubiquitin-proteasome system (30).
Factors that Affect IDE Activity
Zinc stimulates IDE enzyme activity, while copper, alu-
minium, and nitric oxide hinder it (45). In mice, knock-
ing down the IDE gene causes hyperinsulinemia and
insulin counteraction. In Diet-Induced Obese (DIO) mice,
pioglitazone increased IDE activity that declared a mod-
ern algorithm of thiazolidinedione act in insulin resist-
ance treatment. The data indicated that the high-fat diet
enhanced the IDE activity in the liver of DIO mice and
IDE’s movement control by substrate-specific inhibitors
(46, 47). Pioglitazone induced IDE’s movement, implying
a novel medicinal effect in the treatment of T2D.
Induction of IDE Protein by Glucagon and Forskolin
The cells are processed with forskolin, a small molecule
protein kinase A (PKA) activator, which showed a similar
model of IDE protein completion that shows the gluca-
gon promoted IDE protein via the cAMP/PKA signaling
pathway. Forskolin’s PKA pathway had the analogous gain
as glucagon. As a result, the cAMP pathway is probably
obstructed in glucagon-induced IDE upregulation (47).
Inhibition of IDE Protein by TNF‑α
TNF-α raised IDE protein moderately at one hour, then dropped
dramatically after that in the 8-h treatment. In hepatocytes,
TNF-α decreased IDE mRNA. TNF-α seems to have a consid-
erable inhibitory effect on IDE expression in hepatocytes (48).
TNF-α increases insulin resistance in adipocytes and peripheral
tissues via affecting insulin signaling via serine phosphoryla-
tion, leading to the development of T2DM (49, 50).
IDE Inhibitors
IDE inhibitors were first discovered in 1949 and allowed
to treat T2DM (51). Mirsky, and his colleagues, identi-
fied IDE endogenous inhibitors from rat tissue extracts
(52). Partly purified insulin-degrading protease function
they entitled insulinase, the precise identification of these
inhibitors remained unknown. Nevertheless, Mirsky and col-
leagues employed functional properties of IDE inhibitors to
evaluate the hypothesis that IDE antagonists might be an
alternative to treat diabetes inhibit the endogenous insulin
breakdown rather than injected exogenous insulin (53). A
recent study has been found a physiologically active mac-
rocycle that carefully inhibits the IDE in diabetes mellitus
patients, potentially providing fresh therapy alternatives.
The inhibitor significantly enhanced glucose tolerance and
slowed stomach emptying in both lean and obese animals.
The inhibitor modulated circulation levels of glucagon and
amylin, two hormones that are regulated by glucose and
blood sugar levels, in the view of collection to increase insu-
lin levels in mice. The anti-diabetic action of IDE inhibitors
is mediated by several hormones, according to the original
article (54).
13

It has been reported that non-proteinaceous insulinase
inhibitors exhibited hypoglycemic effects both in rabbit and
rodent models (53). Taken together, several antidiabetic drug
moieties have been developed since the 1950s (55–57).
Several studies have been conducted to develop nontarget
specific IDE used like thiol-alkylating compounds, zinc-chela-
tors, cyclic peptide bacitracin, and cyclic peptide bacitracin to
access the functional effect of IDE in insulin catabolism before
the development of selective IDE inhibitors. Non-specific IDE
inhibitors suppress insulin degradation in rat L6 myoblasts,
mouse BC3H1 muscle cells, and HepG2 cells (58–60).
Although robust and rationally selective, Ii1 owned
the drawback of synthesizing. Leissring and colleagues
developed a series of truncated variants, including a 455-
Da allyl ester comprising only the 2-Nap and Arg resi-
dues to achieve the goal (61). Surprisingly, these modi-
fications exhibited unusual substrate selectivity, turning
in potency by up to 300-fold for various substrates, not
existing actives it directed compounds (61). Abdul-Hay,
et al. (61) reported Peptide Hydroxamate (MW > 740) is
target specific IDE inhibitor, which is difficult for chemical
synthesis. This study revealed that, in at least a few cases,
the inhibition of IDE might depend on tertiary interac-
tions amongst the enzyme, substrate, and inhibitor, lend-
ing assistance to the feasibility of uncovering substrate-
selective drug moieties (Table I).
Effect of Hepatic IDE on Insulin
Insulin dictation of IDE operation is intriguing since fat
causes hyperinsulinemia. The relationship is built on the dif-
ferent systems. To find the answer to this query, a research
investigation looked at IDE’s movement in 1c1c7 cells after
pushing the insulin. For 8 h, the cells were given 200 nM
insulin. In both protein and mRNA, IDE activity was meas-
ured. In the insulin-treated cells, no significant change in IDE
expression was found, implying that insulin does not affect the
straightway of IDE’s expression in hepatocytes (48). Under
a liver cell model, researchers reported insulin regulates IDE
activity and this regulation is lost in high glucose circum-
stances. This disruption in IDE protein expression or splicing
cannot be explained by corresponding changes. Furthermore,
it has been found that insulin engagement is associated with
the rise in IDE mRNA levels as well as a trend toward higher
IDE protein levels in high glucose circumstances (62).
ATP Inhibits Insulin‑Degrading Enzyme Activity
The ability of ATP to suppress the IDE enzyme’s degrading
activity in vitro was investigated. Several chromatographic
methods apply to purify the enzyme from rat skeletal mus-
cle. In a polyacrylamide gel, the final purification stage
revealed two bands of 110 and 60 kDa. Insulin breakdown
13
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activity, substratum opposition of unlabeled to labeled insu-
lin, enzyme inhibitor outline, and visualization by a par-
ticular antibody were all used to characterize the enzyme.
Insulin breakdown was inhibited dose-dependently by 1 to
5 mM ATP (Trichloroacetic acid determined by precipitation
and insulin antibody binding). 3 mM adenosine 5-diphos-
phate, adenosine 5-monophosphate, guanosine 5-triphos-
phate, pyrophosphate, −methyleneadenosine 5-triphosphate,
adenosine 5-O-(3thiotriphosphate), and dibutiryl cyclic
adenosine 5-monophosphate inhibited 74%, 4%, 38%, 46%,
65%, 36%, respectively and because the plot of 1/v (insu-
lin degradation) versus ATP concentration non-linear and
the Hill coefficient was greater than one, a kinetic study of
ATP inhibition suggested an allosteric impact (1.51 and
2.44) (Fig. 3). The allosteric inhibition binding constant
was Ki T = 1.5107  M, indicating a decrease in enzyme
affinity caused by ATP. We concluded that ATP inhibits the
enzyme’s insulin breakdown activity (63).
Role of Metal Ions in IDE
Biometals like copper, aluminium, and zinc play an impor-
tant part including Alzheimer’s disease and diabetes melli-
tus. Zinc stimulates IDE enzyme activity, while copper, alu-
minium, and nitric oxide hinder it. In mice, knocking down
the IDE gene causes hyperinsulinemia and insulin resistance
(35, 64, 65). The findings of several experimental techniques
reveal that copper (I) inhibits IDE irreversibly, but that it can
still process its substrates when linked to copper (II) (66).
Metal ions have the potential to influence IDE proteolytic
activity in at least three different ways: Metal ions can trig-
ger conformational changes on IDE substrates, which can
result in proteolytic resistance forms. In this regard, zinc,
but not copper, has been shown to cause the development
of A aggregates that are resistant to IDE proteolytic activity
(67, 68). External influences, such as high metal concen-
trations that encourage Aβ aggregate formation, may con-
tribute to Aβ buildup by lowering the peptide’s sensitivity
to proteolytic degradation; ii. Direct metal ion binding to
IDE may affect the enzyme’s function. Metal ions have long
been known to help enzymes stay stable and active (69). The
binding of metal cofactors in various oxidation states, such
as calcium, magnesium, manganese, copper, zinc, and iron,
has been demonstrated to boost the thermal stabilities of
proteins (70) and to have a significant impact on their struc-
ture and activity (71). Modulation of IDE’s proteolytic activ-
ity could thus occur via a variety of mechanisms, including
exogenous ligand coordination, substitution, or removal
of the catalytic metal (72). Furthermore, metals may bind
to residues in the active site, preventing substrate interac-
tion, or to residues outside the active site, compromising
the enzyme’s structural integrity (71); iii. In vivo, several
metals may influence IDE expression and localization (66).
Pharmaceutical Research

Table I  Structure and common name of insulin degrading enzyme, IDE (Protein/receptor) Potential inhibitor candidates.
Compoun Lipinski’s rule of five
Molecul
d Name / Molecular structure and Referenc
ar Pocket Binding(3D)
ID Interaction with IDE(2D) Properties Value es
formula
Number

Molecular weight
(<500 639.1
Da)

LogP (<5) 2

C42 H86
li1 (103)
O3

H-Bond donor (5) 3

H-bond acceptor (<10) 37

Molecular weight
(<500 757.9
Da)

LogP (<5) 3.5

C41H55N
6bK (104)
7O7

H-Bond donor (5) 7

H-bond acceptor (<10) 8

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Table I  (continued)

Molecular weight
(>500 733.9
Da)

LogP (>5) 7.8

C44H55N (105,106
NTE-1
5O5 )

H-Bond donor (5) 3

H-bond acceptor (<10) 7

Molecular weight
(<500 447.5
Da)

LogP (<5) 2.5

BDM4476 C24H22F
(14)
8 N5O3

H-Bond donor (5) 3

H-bond acceptor (<10) 6

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Table I  (continued)

Molecular weight
(<500 378
Da)

LogP (<5) 0

15349090
B35 (107)
4

H-Bond donor (5) 0

H-bond acceptor (<10) 7

Molecular weight
(<500 479.5
Da)

LogP (<5) 1.6

C21H22F
ML345 (108)
N3O5S2

H-Bond donor (5) 0

H-bond acceptor (<10) 9

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Table I  (continued)

Molecular weight
(<500 735.9
Da)

LogP (<5) 8.6

C44H54F
NTE-2 (109)
N5O4

H-Bond donor (5) 3

H-bond acceptor (<10) 7

Molecular weight
(<500 319.9
Da)

LogP (<5) 4.6

Quinoline C18H26C
(110)
2 lN3

H-Bond donor (5) 1

H-bond acceptor (<10) 3

13
Pharmaceutical Research

Table I  (continued)

Molecular weight
(<500 375.4
Da)

LogP (<5) 5.6

C23H15F
Brequinar (111)
2NO2

H-Bond donor (5) 1

H-bond acceptor (<10) 5

Molecular weight
(<500 324.4
Da)

LogP (<5) 2.9

C20H24N
Quinine (112)
2O2

H-Bond donor (5) 1

H-bond acceptor (<10) 4

­ IGPLOT+, Python, and Protein-Ligand Interaction Profiler with corresponding

The study of molecular docking required software names are L
tools.

Fig. 3  Effect of ATP on IDE

on T2D.

13

Role of Metformin in IDE
Several literature findings suggested that in addition to
Aβ synthesis and conveyance, several signalling path-
ways could entangle in metformin’s impact. In both APP
(amyloid precursor protein, Mo/HuAPP695swe) and
PS1(a mutant human presenilin 1) transgenic mice, the
anti-diabetic medication metformin was found to success-
fully alleviate AD symptoms. In the brains of both APP
and PS1 mice, Glucophage arouses AMPK and enhances
IDE. The metformin could activate IDE to ameliorate
Aβ-induced pathology, mainly AD (73).
In older persons, sporadic cerebral amyloid angiopa-
thy (CAA), marked by cerebrovascular Aβ deposits, pro-
duces brain haemorrhages and dementia. Metformin has
been used to treat T2DM patients, and animal and clinical
research have shown that it can help people with Alz-
heimer’s disease (Fig. 4). Metformin reduced the sever-
ity of CAA by increasing Aβ-cleaving IDE expression.
The clinical application of metformin in CAA treatment
may lead to a novel therapeutic strategy, particularly in
patients with T2D (74–76).
Resveratrol Sustains IDE
When compared to the control, resveratrol causes a sig-
nificant increase in Aβ42 fragmentation, indicating the
polyphenol promotes IDE contingent decay of Aβ42 and
its parts. We observed that implying resveratrol and IDE-
based treatment and prevention methods for sporadic AD
may be promising (77).
Somatostatin Modulates Insulin‑Degrading‑Enzyme
Metabolism
Somatostatin (SST) is a substrate and allosteric regulator
of IDE mobility, allowing a synthetic Aβ-peptide to be
Fig. 4  Effect of metformin on
IDE to treat AD.
13
Pharmaceutical Research
processed more efficiently. Memory and learning prob-
lems are linked to somatostatin inanition in the cortex and
hippocampus of Alzheimer’s patients as well as the devel-
opment of Alzheimer’s disease appears to be linked to a
significant decrease of somatostatin in the mouse CA1
hippocampus region. Somatostatin appears to represent a
role of the principal activity of neprilysin via influencing
its expression and synaptic location (78).
IDE Application in some Therapeutic
Diseases
Schematic diagram for application of IDE in different
chronic diseases shown in Fig. 5.
Inhibition Compared to Activation of IDE
Recent discoveries raise doubts about the wisdom of shut-
ting down the IDE. The discovery that IDE physically
destroys the amyloid protein which saves up pathologi-
cally in Alzheimer’s disease is one of the most signifi-
cant. Rather than inhibiting IDE, it would be preferable
to activate it, a method that has already been shown to
be useful in mice approaches of the particular disorder.
Furthermore, studies from many animal patterns put sus-
picion on the idea of permanently blocking IDE to cure
diabetics (12).
Despite intensive research into IDE’s role as an insulin
protease, there is little evidence that it plays a significant
impact on hepatic insulin clearance in vivo. The effects
of IDE inhibitors on insulin clearance in vivo, such as the
NTE-1 inhibitor, have not been studied or have shown
no effect. The following are the fundamental proof that
implicates IDE in hepatic endosomal insulin proteolysis:
first, its bluff relation for insulin, implying the selec-
tive enzyme for the hormone; second, the discovery that
cleavage insulin feedback achieved from isolated hepatic
endosomes are similar to those produced by purified
Pharmaceutical Research

IDE; and third, the detection of small pools of IDE in
endosomes (81).
Mechanism of IDE in Liver Regulation
In an 8-h investigation, pioglitazone (5 mM) elevated
IDE protein and mRNA in a time-reliant way in the
mouse hepatoma cell line (Hepa 1c1c7 cells), reveal-
ing the formulation of IDE standing order in the liver.
The presence of a free fatty acid (palmitate 300 mM)
increased IDE’s small molecule but decreased mRNA.
IDE protein was reduced by glucagon and TNF-α. Insu-
lin showed no operation in the same circumstance. We
can conclude that pioglitazone and FFA directly boosted
IDE activity in hepatocytes, but Glucagon and TNF-α
reduced it (43, 48).
insulin clearance function. Insulin clearance is mediated by
IDE, which is a significant rate-limiting enzyme. In gene
knockout mice, inactivation of IDE activity causes hyperin-
sulinemia. Insulin and numerous intermediate-size peptides
are degraded by IDE. In diabetic patients, both insulin clear-
ance and IDE activity are diminished, while the cause-effect
link in humans is unknown (81).
Because of IDE’s parts in insulin clearance, IDE inhibi-
tors are utilized to control insulin levels in T2D patients
(83). IDE created certain specific insulin fragments that
have been found to have peculiar biological effects that are
not evident for the entire hormone under the experimental
settings used. The optimum IDE inhibitor for treating T2D
should prioritize insulin and amylin clearance while leav-
ing other IDE substrates, such as glucagon, unaffected (17).

Effect of IDE on Diabetics Treatment and Insulin Advance Prospect of IDE against Alzheimer’s

Clearance Disease (AD)

The liver is an important part of insulin clearance, as it is a. Gene therapy. Insulin signalling and IDE levels are
responsible for eliminating 70% of the insulin produced on reduced in mice fed a high-fat diet, indicating a physi-
the islet (82). Hyperinsulinemia is caused by a decrease in ological relationship in an AD animal model. Increased

Fig. 5  Schematic diagram for

application of IDE in differ-
ent chronic diseases. T2DM,
insulin resistance, obesity,
and neurodegeneration are all
symptoms of T2DM. In both
T2DM and AD, metabolic
excess, chronic inflammation,
and oxidative stress cause cel-
lular dysfunction. Because brain
IR can arise without diabetes,
it’s possible that Alzheimer’s
disease develops in the early
phases of insulin resistance.
Chronic inflammation and
oxidative stress are thought to
be two main mechanisms that
link diabetes and Alzheimer’s
disease (79, 80).

13

Fig. 6  IDE inhibitor prevents disruption of different proteins.
IDE gene dosage has been shown to prevent and treat
the development of AD pathology in a transgenic
mouse model (84, 85). As gene therapy biotechnology
advances, this strategy could become a viable treatment
option for AD. These findings suggest that methods to
improve insulin signalling could be employed as a treat-
ment for Alzheimer’s disease (20, 86). Furthermore, IDE
increasing insulin signalling, and increasing IDE would
avoid the issue of lower insulin neurotrophic effects
induced by elevated IDE (23, 87).
b. Induction of enzymes. Hersh’s team discovered a new
peptide that stimulates IDE activity. Other research
groups are looking for comparable compounds as well.
This method is less invasive and maybe the best strategy
to prevent Alzheimer’s disease.
c. T2D treatment. At present urgent demand is required
against insulin resistance for the subset of T2D patients
with hyperinsulinemia rather than taking exogenous
insulin. Reduced insulin level leads to an increased level
of IDE to initiate degradation, lowering the likelihood
of Alzheimer’s disease (88).
d. Increasing the PPAR/AMPK signalling pathway to
treat T2D and AD. The activators of the PPAR/AMPK
signalling pathway dramatically raised the expression
level of IDE in mice with T2D and AD. They reduced
the buildup of A40 and A42, as well as the impairments
in spatial learning and recognition. To some extent,
PPAR and AMPK activators could be used to treat peo-
ple with T2D and Alzheimer’s disease (54).
IDE for Tumor Suppressor and Anti‑Inflammatory
Protein
In 1994, the researcher hypothesized that IDE inhibits malig-
nant transformation by degrading insulin, preventing this main
growth-stimulatory hormone from attaching to the retinoblas-
toma tumor suppressor protein (RB) and so inactivating it.
13
Pharmaceutical Research
They uncovered a carboxyterminal RB amino acid sequence
that resembled the catalytic core of IDE ten years later. This
structural similarity indicates that insulin degradation is a
common tumor-suppressive mechanism in both RB and IDE.
Following that, a first immunohistochemistry examination of
the differential expression of human IDE in normal tissues,
primary tumors, and their associated lymph node metastases
backed up the initial hypothesis that IDE was an anticancer
molecule. According to this study, IDE contains ankyrin
repeat-like amino acid sequences through which it might bind
and so inhibit the pro-inflammatory factor NF-kappaB and
cyclin-dependent kinases (CDKs). As seen above, IDE also
has two RXL cyclin-binding domains, which could contribute
to its supposed suppression of CDKs. These new findings
imply that IDE may decrease inflammation and oncogenesis
through a variety of pathways that ensure RB function.
This finding suggests a precise method for regulating IDE
activity and/or that there are yet-to-be-identified IDE natural
inhibitors that could be misregulated, at least in this tumor.
Nestin, a type VI intermediate filament protein expressed
by numerous cell types throughout development and at high
levels by tumor cells such as neuroblastoma and glioma
cells, has been discovered to prevent monomeric ubiquitin
breakdown by IDE (30, 89, 90).
IDE Is Involved in Ubiquitin Clearance and Varicella
Zoster Virus Infection
Chickenpox and frequently reactivates in adulthood is
caused by the varicella-zoster virus (VZV) virus, a neu-
rotropic human herpes virus and frequently reactivates in
adulthood, resulting in zoster or, less commonly, cerebral
vasculitis. There have been studies linking IDE to the
etiology of various classification of physiopathological
processes, including diseases, in the last decade, includ-
ing associating the protein with a wide range of cellular
processes, including VZV infection, steroid receptor sign-
aling, and insulin-dependent proteasome modulation. It’s
Pharmaceutical Research
also been suggested that IDE plays a role in rat muscle
cell differentiation and development, implying that it has
various cellular growth and tissue homeostasis functions.
Furthermore, recent discoveries suggest that (i) IDE is
involved in the process of antigenic peptides presented
by class I MHC via a proteasome-independent pathway,
and (ii) ubiquitin can be destroyed by IDE “in vitro,” even
though this last conclusion is somewhat contradictory.
Finally, preliminary research illustrated that in neural
cells attacked with viruses capable of shutting down cel-
lular proteins, the content of IDE was either preserved
or even increased, demonstrating HSP-like action, with
an HSP-like behavior, IDE content was conserved or
even increased. All of these findings point to IDE being
engaged in ubiquitin-proteasome pathways, and hence in
the response related to heat shock (HSR), compelling us
to look into this possibility (30).
The IDE Plays Multifunctional Protein
Many physiologically relevant amyloidogenic peptides,
including insulin, IGF-II, TGF-, glucagon, amylin, atrial
natriuretic peptide (ANP), calcitonin, β-endorphin, A b,
amyloid Bri (ABri), amyloid Dan (ADan), and the C-ter-
minal domain of A b precursor protein (APP-AICD),
somatostatin, are reduced mono shown in Fig. 6 (8, 46). As
a result, poisonous aggregates are prevented from forming,
leading to a variety of clinical disorders (91–93).
However, data suggests that IDE functions not just
through proteolysis but also interactions with other
intracellular proteins (94, 95), containing amyloido-
genic peptides with chaperone-like function (96, 97) and
α-synuclein (α-syn), an amyloidogenic peptide involved in
Parkinson’s disease (98, 99). By binding to androgen and
glucocorticoid receptors and boosting their DNA binding
capacity, IDE may have a role in the cross-talk between
insulin/insulin-like growth factors and steroid hormone
signalling pathways (100).
Coronary Heart Disease
In the Chinese Han population, IDE gene polymorphisms
are strongly linked to the likelihood of developing coronary
heart disease (CHD). Caravaggio, et al. (101) investigated
Ldlr−/− mice and found that the reduced IDE levels in the
mice increased the levels of Aβ and AGEs, thus inducing
inflammation and atherosclerosis. This clinical study found
that IDE gene polymorphisms were linked to the develop-
ment of atherosclerotic lesions, including CHD (102).
Conclusion
The IDE is a multifunctional protein that leads initiate sev-
eral pathological conditions in humans. The therapeutic
strategies targeting IDE could be effective in treating sev-
eral diseases. IDE breaks down insulin and amylin, both
of which are linked to T2D, as well as amyloid-β(Aβ), a
peptide involved in Alzheimer’s disease pathology. Hyper-
insulinemia and T2D develop Alzheimer’s disease in the
senior citizen due to enzyme activity, including substrate
affinities. We believe that a lack of IDE causes an increase in
Aβ and thus AD pathology in the brain. Although the study
was conducted in liver hepatocytes, the findings may apply
to IDE control in other tissues including kidneys, brain, and
adipose tissues. Whereas, emerging IDE inhibitors against
T2D, and AD are in the earlier drug discovery stage, reveal-
ing IDE structure and function will illustrate new insight
into drug discovery to treat IDE-related diseases.
ACKNOWLEDGMENTS AND DISCLOSURES  We thank Dr. Dang Fabin
from Beth Israel Deaconess Medical Center (BIDMC), Harvard for
the critical reading of this manuscript.
Authors’ Contribution  Md. Shofiul Azam conceived the idea and
wrote the original manuscript, writing review and editing. Md. Wahi-
duzzaman analyzed the molecular docking, writing review, and edit-
ing, Md. Reyad-ul-Ferdous, Md. Nahidul Islam, and Mukta Roy
writing review and editing. All authors have read and agreed to the
current version of the manuscript.
Data Availability  Protein and Ligand ID accession numbers were
collected from CRSB (Protein Data Bank) and PubChem databases
respectively. Molecular docking study required software names are
LIGLOT+, Python, and Protein-Ligand Interaction Profiler with cor-
responding tools.
Declarations 
Ethics Approval and Consent to Participate  This study does not relate
to any human and animal trial.
Consent for Publication  The author declares no conflict of interest.
Competing Interests  Not applicable.
References
1. Malito E, Hulse RE, Tang WJ. Amyloid beta-degrading crypti-
dases: insulin degrading enzyme, presequence peptidase, and
neprilysin. Cell Mol Life Sci. 2008;65(16):2574–85.
2. Hersh LB. The insulysin (insulin degrading enzyme) enigma.
Cell Mol Life Sci. 2006;63(21):2432–4.
3. Fernández-Gamba A, Leal MC, Morelli L, Castaño EM.
Insulin-degrading enzyme: structure-function relationship
13

and its possible roles in health and disease. Curr Pharm Des.
2009;15(31):3644–55.
4. Kurochkin IV, Goto S. Alzheimer's beta-amyloid peptide specifi-
cally interacts with and is degraded by insulin degrading enzyme.
FEBS Lett. 1994;345(1):33–7.
5. Guo Q, Manolopoulou M, Bian Y, Schilling AB, Tang WJ.
Molecular basis for the recognition and cleavages of IGF-II,
TGF-alpha, and amylin by human insulin-degrading enzyme. J
Mol Biol. 2010;395(2):430–43.
6. Fo A, Cameron PH, Cm M, Kouach M, Briand G. Endosomal
proteolysis of glucagon at neutral pH generates the bioactive
degradation product Miniglucagon-(19–29). Endocrinology.
2003;144(12):5353–64.
7. Bennett RG, Hamel FG, Duckworth WC. An insulin-degrad-
ing enzyme inhibitor decreases amylin degradation, increases
amylin-induced cytotoxicity, and increases amyloid formation
in insulinoma cell cultures. Diabetes. 2003;52(9):2315–20.
8. Ciaccio C, Tundo GR, Grasso G, Spoto G, Marasco D, Ruvo M,
Gioia M, Rizzarelli E, Coletta M. Somatostatin: a novel substrate
and a modulator of insulin-degrading enzyme activity. J Mol
Biol. 2009;385(5):1556–67.
9. Duckworth WC, Bennett RG, Hamel FG. Insulin degradation:
progress and potential. Endocr Rev. 1998;19(5):608–24.
10. Leissring MA, Farris W, Wu X, Christodoulou DC, Haigis MC,
Guarente L, Selkoe DJ. Alternative translation initiation gener-
ates a novel isoform of insulin-degrading enzyme targeted to
mitochondria. Biochem J. 2004;383(Pt. 3):439–46.
11. Farris W, Leissring MA, Hemming ML, Chang AY, Selkoe DJ.
Alternative splicing of human insulin-degrading enzyme yields
a novel isoform with a decreased ability to degrade insulin and
amyloid β-protein. Biochemistry. 2005;44(17):6513–25.
12. Leissring MA, Farris W, Chang AY, Walsh DM, Wu X, Sun X,
Frosch MP, Selkoe DJ. Enhanced proteolysis of beta-amyloid
in APP transgenic mice prevents plaque formation, secondary
pathology, and premature death. Neuron. 2003;40(6):1087–93.
13. Leissring MA, Selkoe DJ. Structural biology: enzyme target to
latch on to. Nature. 2006;443:761+.
14. Deprez-Poulain R, Hennuyer N, Bosc D, Liang WG, Enée E,
Marechal X, Charton J, Totobenazara J, Berte G, Jahklal J,
Verdelet T, Dumont J, Dassonneville S, Woitrain E, Gauriot
M, Paquet C, Duplan I, Hermant P, Cantrelle F-X, et al. Cata-
lytic site inhibition of insulin-degrading enzyme by a small
molecule induces glucose intolerance in mice. Nat Commun.
2015;6(1):8250.
15. Fakhrai-Rad H, Nikoshkov A, Kamel A, Fernström M, Zierath
JR, Norgren S, Luthman H, Galli J. Insulin-degrading enzyme
identified as a candidate diabetes susceptibility gene in GK rats.
Hum Mol Genet. 2000;9(14):2149–58.
16. Noinaj N, Song E-S, Bhasin S, Alper B, Schmidt W, Hersh L,
Rodgers D. Anion activation site of insulin-degrading enzyme.
J Biol Chem. 2011;287:48–57.
17. Tang W-J. Targeting insulin-degrading enzyme to treat type 2
diabetes mellitus. Trends Endocrinol Metab. 2016;27(1):24–34.
18. Jabeen K, Rehman K, Akash MSH. Genetic mutations of
APOEε4 carriers in cardiovascular patients lead to the devel-
opment of insulin resistance and risk of Alzheimer's disease. J
Biochem Mol Toxicol. 2022;36(2):e22953.
19. Rehman K, Akash MSH, Liaqat A, Kamal S, Qadir MI, Rasul A.
Role of Interleukin-6 in development of insulin resistance and
type 2. Diabetes Mellitus. 2017;27(3):229–36.
20. Kurochkin IV, Guarnera E, Berezovsky IN. Insulin-degrading
enzyme in the fight against Alzheimers disease. Trends Pharma-
col Sci. 2018;39(1):49–58.
21. Qiu WQ, Folstein MF. Insulin, insulin-degrading enzyme and
amyloid-beta peptide in Alzheimer's disease: review and hypoth-
esis. Neurobiol Aging. 2006;27(2):190–8.
13
Pharmaceutical Research
22. Carrasquillo MM, Belbin O, Zou F, Allen M, Ertekin-Taner
N, Ansari M, Wilcox SL, Kashino MR, Ma L, Younkin LH,
Younkin SG, Younkin CS, Dincman TA, Howard ME, Howell
CC, Stanton CM, Watson CM, Crump M, Vitart V, et al. Con-
cordant Association of Insulin Degrading Enzyme Gene (IDE)
variants with IDE mRNA, Aß, and Alzheimer's disease. PLoS
One. 2010;5(1):e8764.
23. Zhao L, Teter B, Morihara T, Lim GP, Ambegaokar SS, Ubeda
OJ, Frautschy SA, Cole GM. Insulin-degrading enzyme as
a downstream target of insulin receptor signaling cascade:
implications for Alzheimer's disease intervention. J Neurosci.
2004;24(49):11120–6.
24. Huang W, Ramsey KM, Marcheva B, Bass J. Circadian rhythms,
sleep, and metabolism. J Clin Invest. 2011;121(6):2133–41.
25. Vitaterna MH, Takahashi JS, Turek FW. Overview of circadian
rhythms. Alcohol Res Health. 2001;25(2):85–93.
26. Amata O, Marino T, Russo N, Toscano M. Human insulin-
degrading enzyme working mechanism. J Am Chem Soc.
2009;131(41):14804–11.
27. Leissring MA, Selkoe DJ. Enzyme target to latch on to. Nature.
2006;443(7113):761–2.
28. Shen Y, Joachimiak A, Rosner MR, Tang WJ. Structures of
human insulin-degrading enzyme reveal a new substrate recog-
nition mechanism. Nature. 2006;443(7113):870–4.
29. Elsässer B, Goettig P. Mechanisms of proteolytic enzymes
and their inhibition in QM/MM studies. Int J Mol Sci.
2021;22(6):3232.
30. Tundo GR, Sbardella D, Ciaccio C, Bianculli A, Orlandi A,
Desimio MG, Arcuri G, Coletta M, Marini S. Insulin-degrading
enzyme (IDE): a novel heat shock-like protein. J Biol Chem.
2013;288(4):2281–9.
31. Song ES, Rodgers DW, Hersh LB. Insulin-degrading enzyme is
not secreted from cultured cells. Sci Rep. 2018;8(1):2335.
32. Son SM, Kang S, Choi H, Mook-Jung I. Statins induce insulin-
degrading enzyme secretion from astrocytes via an autophagy-
based unconventional secretory pathway. Mol Neurodegener.
2015;10:56.
33. Galagovsky D, Katz MJ, Acevedo JM, Sorianello E, Glavic A,
Wappner P. The Drosophila insulin-degrading enzyme restricts
growth by modulating the PI3K pathway in a cell-autonomous
manner. Mol Biol Cell. 2014;25(6):916–24.
34. Sharma SK, Chorell E, Steneberg P, Vernersson-Lindahl E,
Edlund H, Wittung-Stafshede P. Insulin-degrading enzyme pre-
vents α-synuclein fibril formation in a nonproteolytical manner.
Sci Rep. 2015;5:12531.
35. Bellia F, Pietropaolo A, Grasso G. Formation of insulin frag-
ments by insulin-degrading enzyme: the role of zinc(II) and cys-
tine bridges. J Mass Spectrometry. 2013;48(2):135–40.
36. Carpenter JE, Jackson W, de Souza GA, Haarr L, Grose C. Insu-
lin-degrading enzyme binds to the nonglycosylated precursor
of varicella-zoster virus gE protein found in the endoplasmic
reticulum. J Virol. 2010;84(2):847–55.
37. Im H, Manolopoulou M, Malito E, Shen Y, Zhao J, Neant-Fery
M, Sun CY, Meredith SC, Sisodia SS, Leissring MA, Tang WJ.
Structure of substrate-free human insulin-degrading enzyme
(IDE) and biophysical analysis of ATP-induced conformational
switch of IDE. J Biol Chem. 2007;282(35):25453–63.
38. Grasso G, Bush AI, D’Agata R, Rizzarelli E, Spoto G. Enzyme
solid-state support assays: a surface plasmon resonance and mass
spectrometry coupled study of immobilized insulin degrading
enzyme. Eur Biophys J. 2008;38(4):407.
39. Hulse RE, Ralat LA, Wei-Jen T. Structure, function, and regula-
tion of insulin-degrading enzyme. Vitam Horm. 2009;80:635–48.
40. Çakir B, Dağliyan O, Dağyildiz E, Bariş İ, Kavakli IH, Kizilel
S, Türkay M. Structure based discovery of small molecules to
Pharmaceutical Research
regulate the activity of human insulin degrading enzyme. PLoS
One. 2012;7(2):e31787.
41. Manolopoulou M, Guo Q, Malito E, Schilling AB, Tang W-J.
Molecular basis of catalytic chamber-assisted unfolding and
cleavage of human insulin by human insulin-degrading enzyme.
J Biol Chem. 2009;284(21):14177–88.
42. Misbin RI, Almira EC, Duckworth WC, Mehl TD. Inhibition of
insulin degradation by insulin-like growth factors. Endocrinol-
ogy. 1983;113(4):1525–7.
43. González-Casimiro CM, Merino B, Casanueva-Álvarez E,
Postigo-Casado T, Cámara-Torres P, Fernández-Díaz CM,
Leissring MA, Cózar-Castellano I, Perdomo G. Modulation of
insulin sensitivity by insulin-degrading enzyme. Biomedicines.
2021;9(1):86.
44. Farris W, Mansourian S, Leissring MA, Eckman EA, Bertram
L, Eckman CB, Tanzi RE, Selkoe DJ. Partial loss-of-function
mutations in insulin-degrading enzyme that induce diabetes also
impair degradation of amyloid &#x3b2;-protein. Am J Pathol.
2004;164(4):1425–34.
45. Leissring MA, Malito E, Hedouin S, Reinstatler L, Sahara T,
Abdul-Hay SO, Choudhry S, Maharvi GM, Fauq AH, Huzarska
M, May PS, Choi S, Logan TP, Turk BE, Cantley LC, Manolo-
poulou M, Tang W-J, Stein RL, Cuny GD, Selkoe DJ. Designed
inhibitors of insulin-degrading enzyme regulate the catabolism
and activity of insulin. PLoS One. 2010;5(5):e10504.
46. Maianti JP, McFedries A, Foda ZH, Kleiner RE, Du XQ,
Leissring MA, Tang WJ, Charron MJ, Seeliger MA, Saghat-
elian A, Liu DR. Anti-diabetic activity of insulin-degrading
enzyme inhibitors mediated by multiple hormones. Nature.
2014;511(7507):94–8.
47. Maianti JP, Tan GA, Vetere A, Welsh AJ, Wagner BK, Seel-
iger MA, Liu DR. Substrate-selective inhibitors that reprogram
the activity of insulin-degrading enzyme. Nat Chem Biol.
2019;15(6):565–74.
48. Wei X, Ke B, Zhao Z, Ye X, Gao Z, Ye J. Regulation of insu-
lin degrading enzyme activity by obesity-associated factors and
pioglitazone in liver of diet-induced obese mice. PLoS One.
2014;9(4):e95399.
49. Akash MSH, Rehman K, Liaqat A. Tumor necrosis factor-alpha:
role in development of insulin resistance and pathogenesis of
type 2 diabetes mellitus. J Cell Biochem. 2018;119(1):105–10.
50. Faheem A, Rehman K, Jabeen K, Akash MSH. Nicotine-medi-
ated upregulation of microRNA-141 expression determines
adipokine-intervened insulin resistance. Environ Toxicol Phar-
macol. 2020;80:103506.
51. Mirsky IA, Broh-Kahn RH. The inactivation of insulin by tissue
extracts; the distribution and properties of insulin inactivating
extracts. Arch Biochem. 1949;20(1):1–9.
52. Mirsky IA, Simkin B, Broh-Kahn RH. The inactivation of insulin
by tissue extracts. VI. The existence, distribution and properties
of an insulinase inhibitor. Arch Biochem. 1950;28(3):415–23.
53. Mirsky IA, Perisutti G. Effect of insulinase-inhibitor on
hypoglycemic action of insulin. Science (New York, NY).
1955;122(3169):559–60.
54. Li H, Wu J, Zhu L, Sha L, Yang S, Wei J, Ji L, Tang X, Mao K,
Cao L, Wei N, Xie W, Yang Z. Insulin degrading enzyme con-
tributes to the pathology in a mixed model of Type 2 diabetes
and Alzheimer's disease: possible mechanisms of IDE in T2D
and AD. Biosci Rep. 2018;38(1):BSR20170862.
55. Mirsky IA, Perisutti G, Gitelson S. The role of insulinase in the
hypoglycemic response to sulfonylureas. Ann N Y Acad Sci.
1957;71(1):103–11.
56. Leites SM, Smirnov NP. The importance of the insulin inactivat-
ing properties of the liver (insulinase) in the mechanism of action
of antidiabetic sulfonamide preparations. Bull Exp Biol Med.
1959;47(6):711–4.
57. Marigo S, Panelli G. Insulinase and its inhibition by hypoglyce-
mic sulfonamides; data on insulin sensitivity during tolbutamide
therapy. Arch Sci Med. 1958;105(6):587–609.
58. Gehm BD, Rosner MR. Regulation of insulin, epidermal growth
factor, and transforming growth factor-alpha levels by growth fac-
tor-degrading enzymes. Endocrinology. 1991;128(3):1603–10.
59. Kayalar C, Wong WT. Metalloendoprotease inhibitors which
block the differentiation of L6 myoblasts inhibit insulin degrada-
tion by the endogenous insulin-degrading enzyme. J Biol Chem.
1989;264(15):8928–34.
60. Kayalar C, Wong WT, Hendrickson L. Differentiation of BC3H1
and primary skeletal muscle cells and the activity of their endog-
enous insulin-degrading enzyme are inhibited by the same metal-
loendoprotease inhibitors. J Cell Biochem. 1990;44(3):137–51.
61. Abdul-Hay SO, Lane AL, Caulfield TR, Claussin C, Bertrand
J, Masson A, Choudhry S, Fauq AH, Maharvi GM, Leissring
MA. Optimization of peptide Hydroxamate inhibitors of insulin-
degrading enzyme reveals marked substrate-selectivity. J Med
Chem. 2013;56(6):2246–55.
62. Pivovarova O, Gögebakan Ö, Pfeiffer AFH, Rudovich N. Glucose
inhibits the insulin-induced activation of the insulin-degrading
enzyme in HepG2 cells. Diabetologia. 2009;52(8):1656–64.
63. Camberos MC, Perez AA, Udrisar DP, Wanderley MI, Cresto JC.
ATP inhibits insulin-degrading enzyme activity. Exp Biol Med.
2001;226(4):334–41.
64. Perlman RK, Rosner MR. Identification of zinc ligands of the
insulin-degrading enzyme. J Biol Chem. 1994;269(52):33140–5.
65. Bellia F, Grasso G. The role of copper(II) and zinc(II) in the
degradation of human and murine IAPP by insulin-degrading
enzyme. J Mass Spectrom. 2014;49(4):274–9.
66. Grasso G, Salomone F, Tundo GR, Pappalardo G, Ciaccio C,
Spoto G, Pietropaolo A, Coletta M, Rizzarelli E. Metal ions
affect insulin-degrading enzyme activity. J Inorg Biochem.
2012;117:351–8.
67. Bellia F, Lanza V, Ahmed IMM, Garcia-Vinuales S, Veiss
E, Arizzi M, Calcagno D, Milardi D, Grasso G. Site directed
mutagenesis of insulin-degrading enzyme allows singling out
the molecular basis of peptidase versus E1-like activity: the role
of metal ions. Metallomics. 2019;11(2):278–81.
68. Ritchie CW, Bush AI, Mackinnon A, Macfarlane S, Mastwyk
M, MacGregor L, Kiers L, Cherny R, Li Q-X, Tammer A, Car-
rington D, Mavros C, Volitakis I, Xilinas M, Ames D, Davis S,
Beyreuther K, Tanzi RE, Masters CL. Metal-protein attenuation
with Iodochlorhydroxyquin (Clioquinol) targeting Aβ amyloid
deposition and toxicity in Alzheimer disease: a pilot phase 2
clinical trial. Arch Neurol. 2003;60(12):1685–91.
69. Hamel FG, Bennett RG, Duckworth WC. Regulation of multi-
catalytic enzyme activity by insulin and the insulin-degrading
enzyme. Endocrinology. 1998;139(10):4061–6.
70. Kim N, Lee HJ. Redox-active metal ions and amyloid-
degrading enzymes in Alzheimer’s disease. Int J Mol Sci.
2021;22(14):7697.
71. Hadden JM, Déclais A-C, Phillips SEV, Lilley DMJ. Metal ions
bound at the active site of the junction-resolving enzyme T7
endonuclease I. EMBO J. 2002;21(13):3505–15.
72. Soulière MF, Perreault JP, Bisaillon M. Characterization of the
vaccinia virus D10 decapping enzyme provides evidence for a
two-metal-ion mechanism. Biochem J. 2009;420(1):27–35.
73. Lu X-Y, Huang S, Chen Q-B, Zhang D, Li W, Ao R, Leung FC-Y,
Zhang Z, Huang J, Tang Y, Zhang S-J. Metformin ameliorates Aβ
pathology by insulin-degrading enzyme in a transgenic mouse
model of Alzheimer’s disease. Oxidative Med Cell Longev.
2020;2020:2315106.
74. Inoue Y, Masuda T, Misumi Y, Ando Y, Ueda M. Metformin
attenuates vascular pathology by increasing expression of
insulin-degrading enzyme in a mixed model of cerebral
13

amyloid angiopathy and type 2 diabetes mellitus. Neurosci Lett.
2021;762:136136.
75. Khalil MR, Ebeid A, Fayed H, Abd-Elhady S. Metformin: new
insights into Alzheimer disease protection. Asian. J Biochem.
2019;15(1):21–7.
76. Rotermund C, Machetanz G, Fitzgerald JC. The therapeutic
potential of metformin in neurodegenerative diseases. Front
Endocrinol. 2018;9:400.
77. Krasinski CA, Ivancic VA, Zheng Q, Spratt DE, Lazo ND. Res-
veratrol sustains insulin-degrading enzyme activity toward Aβ42.
ACS Omega. 2018;3(10):13275–82.
78. Tundo G, Ciaccio C, Sbardella D, Boraso M, Viviani B, Coletta
M, Marini S. Somatostatin modulates insulin-degrading-enzyme
metabolism: implications for the regulation of microglia activity
in AD. PLoS One. 2012;7(4):e34376.
79. Rehman K, Akash MSH. Mechanisms of inflammatory
responses and development of insulin resistance: how are they
interlinked? J Biomed Sci. 2016;23(1):87.
80. Tegg M. Plasma insulin-degrading enzyme: Characterisation
and evaluation as a potential biomarker for Alzheimer's dis-
ease.Masters Thesis. In: School of Medical Sciences: Edith
Cowan University. Joondalup, Australia; 2014.
81. Leissring MA, González-Casimiro CM, Merino B, Suire CN,
Perdomo G. Targeting insulin-degrading enzyme in insulin
clearance. Int J Mol Sci. 2021;22(5):2235.
82. Costes S, Butler PC. Insulin-degrading enzyme inhibition, a novel
therapy for type 2 diabetes? Cell Metab. 2014;20(2):201–3.
83. Nash Y, Ganoth A, Borenstein-Auerbach N, Levy-Barazany H,
Goldsmith G, Kopelevich A, Pozyuchenko K, Sakhneny L, Laz-
don E, Blanga-Kanfi S, Alhadeff R, Benromano T, Landsman
L, Tsfadia Y, Frenkel D. From virus to diabetes therapy: char-
acterization of a specific insulin-degrading enzyme inhibitor for
diabetes treatment. FASEB J. 2021;35(5):e21374.
84. Del Campo M, Stargardt A, Veerhuis R, Reits E, Teunissen CE.
Accumulation of BRI2-BRICHOS ectodomain correlates with a
decreased clearance of Aβ by insulin degrading enzyme (IDE) in
Alzheimer's disease. Neurosci Lett. 2015;589:47–51.
85. Pivovarova-Ramich O, Höhn A, Grune T, Pfeiffer A, Rudovich
N. Insulin-degrading enzyme: new therapeutic target for diabetes
and Alzheimer’s disease? Ann Med. 2016;48:1–11.
86. Zou F, Carrasquillo MM, Pankratz VS, Belbin O, Morgan K,
Allen M, Wilcox SL, Ma L, Walker LP, Kouri N, Burgess JD,
Younkin LH, Younkin SG, Younkin CS, Bisceglio GD, Crook
JE, Dickson DW, Petersen RC, Graff-Radford N, et al. Gene
expression levels as endophenotypes in genome-wide associa-
tion studies of Alzheimer disease. Neurology. 2010;74(6):480–6.
87. Ferdous MR, Azam MS. Cardiac disease: current approaches
to gene therapy. In: Bulletin of Medical and Clinical Research;
2020. p. 62–75.
88. Qiu C, Gelaye B, Denis M, Tadesse MG, Luque Fernandez MA,
Enquobahrie DA, Ananth CV, Sanchez SE, Williams MA. Circa-
dian clock-related genetic risk scores and risk of placental abrup-
tion. Placenta. 2015;36(12):1480–6.
89. Ralat LA, Kalas V, Zheng Z, Goldman RD, Sosnick TR, Tang
WJ. Ubiquitin is a novel substrate for human insulin-degrading
enzyme. J Mol Biol. 2011;406(3):454–66.
90. Joly N, Engl C, Jovanovic G, Huvet M, Toni T, Sheng X, Stumpf
MP, Buck M. Managing membrane stress: the phage shock pro-
tein (Psp) response, from molecular mechanisms to physiology.
FEMS Microbiol Rev. 2010;34(5):797–827.
91. Mukherjee A, Morales-Scheihing D, Butler PC, Soto C. Type
2 diabetes as a protein misfolding disease. Trends Mol Med.
2015;21(7):439–49.
92. Powers ET, Morimoto RI, Dillin A, Kelly JW, Balch WE. Bio-
logical and chemical approaches to diseases of proteostasis defi-
ciency. Annu Rev Biochem. 2009;78:959–91.
13
Pharmaceutical Research
93. Chiti F, Dobson CM. Protein misfolding, functional amyloid, and
human disease. Annu Rev Biochem. 2006;75:333–66.
94. Bennett RG, Fawcett J, Kruer M, Duckworth W, Hamel FG.
Insulin inhibition of the proteasome is dependent on degrada-
tion of insulin by insulin-degrading enzyme. J Endocrinol.
2003;177(3):399–405.
95. Epting CL, King FW, Pedersen A, Zaman J, Ritner C, Bernstein
HS. Stem cell antigen-1 localizes to lipid microdomains and
associates with insulin degrading enzyme in skeletal myoblasts.
J Cell Physiol. 2008;217(1):250–60.
96. Llovera RE, De Tullio M, Alonso LG, Leissring MA, Kaufman
SB, Roher AE, de Prat GG, Morelli L, Castaño EM. The catalytic
domain of insulin-degrading enzyme forms a denaturant-resistant
complex with amyloid β peptide: implications for Alzheimer dis-
ease pathogenesis. J Biol Chem. 2008;283(25):17039–48.
97. de Tullio MB, Castelletto V, Hamley IW, Martino Adami PV,
Morelli L, Castaño EM. Proteolytically inactive insulin-degrad-
ing enzyme inhibits amyloid formation yielding non-neurotoxic
Aβ peptide aggregates. PLoS One. 2013;8(4):e59113.
98. De Kimpe L, Scheper W. From alpha to omega with Aβ: targeting
the multiple molecular appearances of the pathogenic peptide in
Alzheimer's disease. Curr Med Chem. 2010;17(3):198–212.
99. Steneberg P, Bernardo L, Edfalk S, Lundberg L, Backlund
F, Östenson C-G, Edlund H. The type 2 diabetes–associated
gene ide is required for insulin secretion and suppression of
α-synuclein levels in β-cells. Diabetes. 2013;62(6):2004–14.
100. Tundo GR, Sbardella D, Ciaccio C, Grasso G, Gioia M,
Coletta A, Polticelli F, Di Pierro D, Milardi D, Van Endert P,
Marini S, Coletta M. Multiple functions of insulin-degrading
enzyme: a metabolic crosslight? Crit Rev Biochem Mol Biol.
2017;52(5):554–82.
101. Caravaggio JW, Hasu M, MacLaren R, Thabet M, Raizman
JE, Veinot JP, Marcel YL, Milne RW, Whitman SC. Insulin-
degrading enzyme deficiency in bone marrow cells increases ath-
erosclerosis in LDL receptor-deficient mice. Cardiovasc Pathol.
2013;22(6):458–64.
102. Cai P, Zhong W, Jia M, Yu C-q, Peng Y, Wang Y, Wang H-y,
Zeng C, Bai Y, Wang X. The gene polymorphisms of insulin
degrading enzyme ( IDE ) are associated with the risk of coro-
nary heart disease in Chinese Han population. In.; 2016.
103. Rocha NKR, Themoteo R, Brentani H, Forlenza OV, De Paula
VJR. Neuronal-glial interaction in a triple-transgenic mouse
model of Alzheimer's disease: gene ontology and Lithium path-
ways. Front Neurosci. 2020;14:579984.
104. Hahn F, Schmalen A, Setz C, Friedrich M, Schlößer S, Kölle J,
Spranger R, Rauch P, Fraedrich K, Reif T, Karius-Fischer J, Bal-
asubramanyam A, Henklein P, Fossen T, Schubert U. Proteolysis
of mature HIV-1 p6 gag protein by the insulin-degrading enzyme
(IDE) regulates virus replication in an Env-dependent manner.
PLoS One. 2017;12(4):e0174254.
105. Nunez LR, Jesch SA, Gaspar ML, Almaguer C, Villa-Garcia M,
Ruiz-Noriega M, Patton-Vogt J, Henry SA. Cell wall integrity
MAPK pathway is essential for lipid homeostasis. J Biol Chem.
2008;283(49):34204–17.
106. Durham TB, Toth JL, Klimkowski VJ, Cao JX, Siesky AM,
Alexander-Chacko J, Wu GY, Dixon JT, McGee JE, Wang Y,
Guo SY, Cavitt RN, Schindler J, Thibodeaux SJ, Calvert NA,
Coghlan MJ, Sindelar DK, Christe M, Kiselyov VV, et al. Dual
exosite-binding inhibitors of insulin-degrading enzyme challenge
its role as the primary mediator of insulin clearance in vivo. J
Biol Chem. 2015;290(33):20044–59.
107. Wang T, Yang N, Liang C, Xu H, An Y, Xiao S, Zheng M, Liu
L, Wang G, Nie L. Detecting protein-protein interaction based
on protein fragment complementation assay. Curr Protein Pept
Sci. 2020;21(6):598–610.
Pharmaceutical Research

108. Bannister TD, Wang H, Abdul-Hay SO, Masson A, Madoux F,
Ferguson J, Mercer BA, Schurer S, Zuhl A, Cravatt BF, Leis-
sring MA, Hodder P. ML345, a small-molecule inhibitor of the
insulin-degrading enzyme (IDE). In: Probe reports from the NIH
molecular libraries program. Bethesda: National Center for Bio-
technology Information (US); 2010.
109. Escudero MA, Vilanova E. Purification and characterization
of naturally soluble neuropathy target esterase from chicken
sciatic nerve by HPLC and western blot. J Neurochem.
1997;69(5):1975–82.
110. Lindner T, Loktev A, Altmann A, Giesel F, Kratochwil C, Debus
J, Jäger D, Mier W, Haberkorn U. Development of Quinoline-
based Theranostic ligands for the targeting of fibroblast activa-
tion protein. J Nucl Med. 2018;59(9):1415–22.
111. Cuthbertson CR, Guo H, Kyani A, Madak JT, Arabzada Z, Nea-
mati N. The dihydroorotate dehydrogenase inhibitor Brequinar is
synergistic with ENT1/2 inhibitors. ACS Pharmacol Transl Sci.
2020;3(6):1242–52.
112. Allotey-Babington GL, Amponsah SK, Nettey T, Sasu C, Nettey
H. Quinine Sulphate microparticles as treatment for Leishmania-
sis. J Trop Med. 2020;2020:5278518.
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