1Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
JOURNAL VITAE
School of Pharmaceutical and
Food Sciences
ISSN 0121-4004 | ISSNe 2145-2660
University of Antioquia
Medellin, Colombia
Filliations
1Universidad Nacional de Colombia,
Sede Bogotá, Facultad de
Medicina, Departamento de
Ciencias Fisiológicas. Bogotá,
Colombia.
2 Universidad de La Sabana, Facultad de
Medicina. Chía, Colombia.
3 Gimnasio Vermont, Departamento de
Ciencias Naturales – Química IB.
Bogotá, Colombia.
*Corresponding
Jorge Andrés Barrero
jobarreroc@unal.edu.co
Received: 24 August 2021
Accepted: 11 January 2023
Published: 03 February 2023
Fast Skeletal Muscle Troponin and
Tropomyosin as a Dietary Source of
Antidiabetic and Antihypertensive
Bioactive Peptides: An In Silico Study
Troponina y Tropomiosina de Músculo Esquelético como
Fuente Alimentaria de Péptidos Bioactivos Antidiabéticos y
Antihipertensivos: Estudio In Silico
Jorge Andrés Barrero 1,* , María Alejandra Barrero-Casallas2 ,
Angélica María González-Clavijo1 , Marcela Cruz-González3 .
ABSTRACT
Background: The nutraceutical properties of food hydrolysates rely on multiple biochemical
interactions involving the modulation of enzymes and cellular receptors. Numerous bioactive
peptides released from troponin and tropomyosin digestion have been identified. Their
characterization has mostly been performed by hydrolysis catalyzed by proteases unrelated to
the human digestive system. Objective: This study aimed to determine the bioactive profile of
beef, pork, and chicken meat by analyzing the frequency and pharmacokinetics of biopeptides
released from troponin and tropomyosin. Methods: In silico digestion and biopeptide release
frequency were studied by three parameters; bioactive fragments release frequency (A E),
frequency percentage (W), and mean occurrence (A S), all stated on the BIOPEP-UWM platform.
Further on, hydrolysis end-products were screened based on gastrointestinal-absorption
probability and pharmacokinetic profiling performed on SwissADME, SwissTargetPrediction,
and ADME/Tlab bioinformatics web tools. Statistical analyses were performed using a one-way
ANOVA test. Results: Dipeptidyl peptidase-IV (DPP-IV) and angiotensin-converting enzyme
(ACE) inhibiting biopeptides exhibited the highest release frequency. Moreover, W and A S
parameters showed no significant difference (p>0.05) between the myofibrillar isoforms
assessed. Seven biopeptides were classified as highly absorbable and reported optimal
drug-likeness compliance. Although biopeptides hold good pharmacokinetic properties,
the therapeutic potency of biopeptides showed to be lower than those of DPP-IV and ACE-
inhibiting drugs. Conclusions: Troponin and tropomyosin are rich dietary sources of bioactive
peptides, mainly DPP-IV and ACE inhibitors. Digestion end-products are mainly dipeptides
with optimal pharmacokinetic and drug-like properties, suggesting a potential therapeutic
application in hypertensive and hyperglycemic disorders.
Keywords: Bioactive peptides; angiotensin-converting enzyme inhibitors; dipeptidyl-
peptidase IV inhibitors; Tropomyosin; Troponin.
ORIGINAL RESEARCH
Published 03 February 2023
Doi: https://doi.org/10.17533/udea.vitae.v30n1a347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
JOURNAL VITAE
School of Pharmaceutical and
Food Sciences
ISSN 0121-4004 | ISSNe 2145-2660
University of Antioquia
Medellin, Colombia
Filliations
1Universidad Nacional de Colombia,
Sede Bogotá, Facultad de
Medicina, Departamento de
Ciencias Fisiológicas. Bogotá,
Colombia.
2 Universidad de La Sabana, Facultad de
Medicina. Chía, Colombia.
3 Gimnasio Vermont, Departamento de
Ciencias Naturales – Química IB.
Bogotá, Colombia.
*Corresponding
Jorge Andrés Barrero
jobarreroc@unal.edu.co
Received: 24 August 2021
Accepted: 11 January 2023
Published: 03 February 2023
Fast Skeletal Muscle Troponin and
Tropomyosin as a Dietary Source of
Antidiabetic and Antihypertensive
Bioactive Peptides: An In Silico Study
Troponina y Tropomiosina de Músculo Esquelético como
Fuente Alimentaria de Péptidos Bioactivos Antidiabéticos y
Antihipertensivos: Estudio In Silico
Jorge Andrés Barrero 1,* , María Alejandra Barrero-Casallas2 ,
Angélica María González-Clavijo1 , Marcela Cruz-González3 .
ABSTRACT
Background: The nutraceutical properties of food hydrolysates rely on multiple biochemical
interactions involving the modulation of enzymes and cellular receptors. Numerous bioactive
peptides released from troponin and tropomyosin digestion have been identified. Their
characterization has mostly been performed by hydrolysis catalyzed by proteases unrelated to
the human digestive system. Objective: This study aimed to determine the bioactive profile of
beef, pork, and chicken meat by analyzing the frequency and pharmacokinetics of biopeptides
released from troponin and tropomyosin. Methods: In silico digestion and biopeptide release
frequency were studied by three parameters; bioactive fragments release frequency (A E),
frequency percentage (W), and mean occurrence (A S), all stated on the BIOPEP-UWM platform.
Further on, hydrolysis end-products were screened based on gastrointestinal-absorption
probability and pharmacokinetic profiling performed on SwissADME, SwissTargetPrediction,
and ADME/Tlab bioinformatics web tools. Statistical analyses were performed using a one-way
ANOVA test. Results: Dipeptidyl peptidase-IV (DPP-IV) and angiotensin-converting enzyme
(ACE) inhibiting biopeptides exhibited the highest release frequency. Moreover, W and A S
parameters showed no significant difference (p>0.05) between the myofibrillar isoforms
assessed. Seven biopeptides were classified as highly absorbable and reported optimal
drug-likeness compliance. Although biopeptides hold good pharmacokinetic properties,
the therapeutic potency of biopeptides showed to be lower than those of DPP-IV and ACE-
inhibiting drugs. Conclusions: Troponin and tropomyosin are rich dietary sources of bioactive
peptides, mainly DPP-IV and ACE inhibitors. Digestion end-products are mainly dipeptides
with optimal pharmacokinetic and drug-like properties, suggesting a potential therapeutic
application in hypertensive and hyperglycemic disorders.
Keywords: Bioactive peptides; angiotensin-converting enzyme inhibitors; dipeptidyl-
peptidase IV inhibitors; Tropomyosin; Troponin.
ORIGINAL RESEARCH
Published 03 February 2023
Doi: https://doi.org/10.17533/udea.vitae.v30n1a347310
2Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 30 | Number 01 | Article 347310Jorge Andrés Barrero, María Alejandra Barrero-Casallas, Angélica María González-Clavijo, Marcela Cruz-González
INTRODUCTION
Meat is ranked as one of the most important
foodstuffs on a daily-basis diet, being a rich source
of minerals, fatty acids, and both essential and non-
essential amino acids (1). Among the wide range
of polypeptides present within the muscle fibers,
myofibrillar proteins account for more than 50%
of the protein content in meat (2). Two of these
contractile proteins hold significant physiological
functions; troponin, a globular protein with three
subunits (T, I, and C), and tropomyosin, a fibrillar
polypeptide made up of α and β chains. These
myofibrillar proteins are scattered across sarcomeres
along the muscle fibers; thus, besides their well-
established role in muscle contraction, they
represent a major source of dietary amino acids
when digested in the gastrointestinal tract.
Growing research in food chemistry has revealed
the therapeutic effects of nutrients, currently known
as nutraceutical properties (3). Nutraceuticals
exhibit a broad spectrum of health-promoting
properties, including vascular resistance reduction,
glycemic regulation, and free-radicals scavenging,
and have also shown to be beneficial in numerous
metabolic disorders (3). Dietary protein intake,
and myofibrillar proteins, particularly, are a rich
source of biologically active molecules known as
bioactive peptides (4). The biological properties
of these oligopeptides rely on several mechanisms
involving the modulation of enzymes and cellular
receptors. Recent trends in biopeptides research
have led to the development of in silico tools
that enable a simulated/computerized enzymatic
hydrolysis of polypeptides helping to characterize
the bioactive products from protein digestion (5)
relative frequency of release of fragments with a
given activity by selected enzyme(s. As a result, this
bioinformatic approach has allowed researchers to
determine the profile of bioactive peptides released
from dietary protein when hydrolyzed by specific
subsets of enzymes.
As stated by Wang et al. (6), in silico experimentation
has proved to be a useful approach to predict the
bioactive profile, particularly dipeptidyl-peptidase
IV (DPP-IV ) inhibitor y proper ties, of protein
hydrolysates. Although in vivo studies are limited,
they provide valuable evidence that DPP-IV inhibiting
biopeptides from dietary protein intake improve
glycemic regulation and enhance insulin sensitivity
in diabetic animal models (7). Consequently, the
bioactive properties of food-hydrolysates have
been thoroughly investigated, regarding the
pharmacokinetics and drug-likeness of these
nutraceuticals are still scarce (8). Pharmacokinetic
traits must be revealed to elucidate the therapeutic
potential of biopeptides; however, studying the
pharmacological parameters of bioactive peptides
RESUMEN
Antecedentes: Las propiedades nutracéuticas de los hidrolizados de alimentos dependen de múltiples interacciones
bioquímicos que involucran la modulación de enzimas y receptores celulares. Se han identificado numerosos péptidos
bioactivos liberados de la digestión de troponina y tropomiosina, pero su caracterización se ha llevado a cabo principalmente
por hidrólisis catalizada por proteasas ajenas al sistema digestivo humano. Objetivo: Este estudio tuvo como objetivo
determinar el perfil bioactivo de la carne de res, cerdo y pollo mediante el análisis de la frecuencia y farmacocinética de
los biopéptidos liberados de la troponina y la tropomiosina. Métodos: Se estudió la digestión in silico y la frecuencia de
liberación de biopéptidos mediante dos parámetros; frecuencia de liberación de fragmentos bioactivos (A E), frecuencia
porcentual (W) y ocurrencia media (A S ), ambos indicados en la plataforma BIOPEP-UWM. Más adelante, los productos finales
de la hidrólisis se examinaron en función de la probabilidad de absorción gastrointestinal y el perfil farmacocinético realizado
en las herramientas bioinformáticas SwissADME, SwissTargetPrediction y ADME/Tlab. El análisis estadístico se llevó a cabo
mediante una prueba ANOVA de una vía. Resultados: Los biopéptidos inhibidores de la dipeptidil peptidasa IV (DPP-IV) y
la enzima convertidora de angiotensina (ECA) exhibieron la mayor frecuencia de liberación. Además, los parámetros W y A S
no mostraron diferencias significativas (p> 0.05) entre las isoformas miofibrilares evaluadas. Siete biopéptidos se clasificaron
como altamente absorbibles e informaron un cumplimiento óptimo de similitud con el fármaco. Aunque los biopéptidos tienen
propiedades farmacocinéticas adecuadas, su potencia terapéutica demostró ser menor que la de los fármacos inhibidores
de la DPP-IV y la ACE. Conclusiones: La troponina y la tropomiosina son una fuente dietética rica en péptidos bioactivos,
principalmente DPP-IV e inhibidores de la ACE. Los productos finales de la digestión son principalmente dipéptidos con
propiedades farmacocinéticas óptimas y similares a la de los fármacos, lo que sugiere una aplicación terapéutica factible en
trastornos hipertensivos e hiperglicémicos.
Palabras clave: Péptidos bioactivos; Inhibidores de la enzima convertidora de angiotensina; inhibidores de la dipeptidil-
peptidasa IV; Tropomiosina; Troponina.
INTRODUCTION
Meat is ranked as one of the most important
foodstuffs on a daily-basis diet, being a rich source
of minerals, fatty acids, and both essential and non-
essential amino acids (1). Among the wide range
of polypeptides present within the muscle fibers,
myofibrillar proteins account for more than 50%
of the protein content in meat (2). Two of these
contractile proteins hold significant physiological
functions; troponin, a globular protein with three
subunits (T, I, and C), and tropomyosin, a fibrillar
polypeptide made up of α and β chains. These
myofibrillar proteins are scattered across sarcomeres
along the muscle fibers; thus, besides their well-
established role in muscle contraction, they
represent a major source of dietary amino acids
when digested in the gastrointestinal tract.
Growing research in food chemistry has revealed
the therapeutic effects of nutrients, currently known
as nutraceutical properties (3). Nutraceuticals
exhibit a broad spectrum of health-promoting
properties, including vascular resistance reduction,
glycemic regulation, and free-radicals scavenging,
and have also shown to be beneficial in numerous
metabolic disorders (3). Dietary protein intake,
and myofibrillar proteins, particularly, are a rich
source of biologically active molecules known as
bioactive peptides (4). The biological properties
of these oligopeptides rely on several mechanisms
involving the modulation of enzymes and cellular
receptors. Recent trends in biopeptides research
have led to the development of in silico tools
that enable a simulated/computerized enzymatic
hydrolysis of polypeptides helping to characterize
the bioactive products from protein digestion (5)
relative frequency of release of fragments with a
given activity by selected enzyme(s. As a result, this
bioinformatic approach has allowed researchers to
determine the profile of bioactive peptides released
from dietary protein when hydrolyzed by specific
subsets of enzymes.
As stated by Wang et al. (6), in silico experimentation
has proved to be a useful approach to predict the
bioactive profile, particularly dipeptidyl-peptidase
IV (DPP-IV ) inhibitor y proper ties, of protein
hydrolysates. Although in vivo studies are limited,
they provide valuable evidence that DPP-IV inhibiting
biopeptides from dietary protein intake improve
glycemic regulation and enhance insulin sensitivity
in diabetic animal models (7). Consequently, the
bioactive properties of food-hydrolysates have
been thoroughly investigated, regarding the
pharmacokinetics and drug-likeness of these
nutraceuticals are still scarce (8). Pharmacokinetic
traits must be revealed to elucidate the therapeutic
potential of biopeptides; however, studying the
pharmacological parameters of bioactive peptides
RESUMEN
Antecedentes: Las propiedades nutracéuticas de los hidrolizados de alimentos dependen de múltiples interacciones
bioquímicos que involucran la modulación de enzimas y receptores celulares. Se han identificado numerosos péptidos
bioactivos liberados de la digestión de troponina y tropomiosina, pero su caracterización se ha llevado a cabo principalmente
por hidrólisis catalizada por proteasas ajenas al sistema digestivo humano. Objetivo: Este estudio tuvo como objetivo
determinar el perfil bioactivo de la carne de res, cerdo y pollo mediante el análisis de la frecuencia y farmacocinética de
los biopéptidos liberados de la troponina y la tropomiosina. Métodos: Se estudió la digestión in silico y la frecuencia de
liberación de biopéptidos mediante dos parámetros; frecuencia de liberación de fragmentos bioactivos (A E), frecuencia
porcentual (W) y ocurrencia media (A S ), ambos indicados en la plataforma BIOPEP-UWM. Más adelante, los productos finales
de la hidrólisis se examinaron en función de la probabilidad de absorción gastrointestinal y el perfil farmacocinético realizado
en las herramientas bioinformáticas SwissADME, SwissTargetPrediction y ADME/Tlab. El análisis estadístico se llevó a cabo
mediante una prueba ANOVA de una vía. Resultados: Los biopéptidos inhibidores de la dipeptidil peptidasa IV (DPP-IV) y
la enzima convertidora de angiotensina (ECA) exhibieron la mayor frecuencia de liberación. Además, los parámetros W y A S
no mostraron diferencias significativas (p> 0.05) entre las isoformas miofibrilares evaluadas. Siete biopéptidos se clasificaron
como altamente absorbibles e informaron un cumplimiento óptimo de similitud con el fármaco. Aunque los biopéptidos tienen
propiedades farmacocinéticas adecuadas, su potencia terapéutica demostró ser menor que la de los fármacos inhibidores
de la DPP-IV y la ACE. Conclusiones: La troponina y la tropomiosina son una fuente dietética rica en péptidos bioactivos,
principalmente DPP-IV e inhibidores de la ACE. Los productos finales de la digestión son principalmente dipéptidos con
propiedades farmacocinéticas óptimas y similares a la de los fármacos, lo que sugiere una aplicación terapéutica factible en
trastornos hipertensivos e hiperglicémicos.
Palabras clave: Péptidos bioactivos; Inhibidores de la enzima convertidora de angiotensina; inhibidores de la dipeptidil-
peptidasa IV; Tropomiosina; Troponina.
3Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
would require extensive laboratory research and
clinical trials. Fortunately, bioinformatic-assisted
experimental designs allow the evaluation of
compounds based on molecular and topological
descriptors (9); for instance, absorption, distribution,
excretion, metabolism, and toxicity (ADEM/T)
(10) and target prediction analysis (11) enable to
develop of a pharmacokinetic characterization from
a chemoinformatic theoretical approach which help
to elucidate biopeptides pharmacological traits.
Many bioactive peptides have been identified, and
their isolation has been performed by enzymatic
proteolysis catalyzed by a wide range of proteases
unrelated to the human digestive system. This
study aimed to characterize the bioactive peptides
obtained from beef, pork, and chicken meat when
subjected to an in silico digestion by pepsin (gastric
digestion), trypsin, and chymotrypsin (pancreatic
digestion). We expected to identify the difference
in biopeptide-release frequency between the
isoforms of these species and filter the obtained
bioactive peptides based on their drug-likeness
compliance and pharmacokinetic profile. Screening
methods carried out through an in silico approach
serve to identify suitable meat-hydrolysates with
a therapeutic potential before in vitro and in vivo
experimentation.
MATERIALS AND METHODS
Sequence retrieval
Aiming to analyze the bioactive peptides released
from beef (cow - Bos taurus), chicken meat (Gallus
gallus), and pork (pig - Sus scrofa) meat digestion,
the sequences from fast skeletal muscle troponin
subunits (troponin C, troponin T, and troponin I)
and tropomyosin subunits (tropomyosin-α chain
and tropomyosin-β chain) were retrieved from the
UniProt database (https://www.uniprot.org) (12). For
each species, five peptidic sequences were obtained
(Table 1).
Table 1. Data from the proteins retrieved from the UniProt database.
Organism Protein Amino acids length Entry code (ID)
Bos taurus
Troponin C 161 Q148C2|Q148C2_BOVIN
Troponin T 270 Q8MKI3|TNNT3_BOVIN
Troponin I 178 F6QIC1|F6QIC1_BOVIN
Tropomyosin-α 284 Q5KR49|TPM1_BOVIN
Tropomyosin-β 284 Q5KR48|TPM2_BOVIN
Gallus gallus
Troponin C 163 P02588|TNNC2_CHICK
Troponin T 263 P12620|TNNT3_CHICK
Troponin I 183 P68246|TNNI2_CHICK
Tropomyosin-α 284 P04268|TPM1_CHICK
Tropomyosin-β 284 P19352|TPM2_CHICK
Sus scrofa
Troponin C 159 P02587|TNNC2_PIG
Troponin T 271 Q75NG9|TNNT3_PIG
Troponin I 182 Q4JH15|Q4JH15_PIG
Tropomyosin-α 248 A0A4X1TTA0|A0A4X1TTA0_PIG
Tropomyosin-β 287 A1X899|A1X899_PIG
Bioactivity profiling
In silico enzymatic digestion and bioactivity profiling
were analyzed using the BIOPEP-UWM database (13)
especially on these derived from foods and being
constituents of diets that prevent development
of chronic diseases. The database is continuously
updated and modified. The addition of new
peptides and the introduction of new information
about the existing ones (e.g., chemical codes
and references to other databases) and served to
evaluate troponin and tropomyosin as a source of
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
would require extensive laboratory research and
clinical trials. Fortunately, bioinformatic-assisted
experimental designs allow the evaluation of
compounds based on molecular and topological
descriptors (9); for instance, absorption, distribution,
excretion, metabolism, and toxicity (ADEM/T)
(10) and target prediction analysis (11) enable to
develop of a pharmacokinetic characterization from
a chemoinformatic theoretical approach which help
to elucidate biopeptides pharmacological traits.
Many bioactive peptides have been identified, and
their isolation has been performed by enzymatic
proteolysis catalyzed by a wide range of proteases
unrelated to the human digestive system. This
study aimed to characterize the bioactive peptides
obtained from beef, pork, and chicken meat when
subjected to an in silico digestion by pepsin (gastric
digestion), trypsin, and chymotrypsin (pancreatic
digestion). We expected to identify the difference
in biopeptide-release frequency between the
isoforms of these species and filter the obtained
bioactive peptides based on their drug-likeness
compliance and pharmacokinetic profile. Screening
methods carried out through an in silico approach
serve to identify suitable meat-hydrolysates with
a therapeutic potential before in vitro and in vivo
experimentation.
MATERIALS AND METHODS
Sequence retrieval
Aiming to analyze the bioactive peptides released
from beef (cow - Bos taurus), chicken meat (Gallus
gallus), and pork (pig - Sus scrofa) meat digestion,
the sequences from fast skeletal muscle troponin
subunits (troponin C, troponin T, and troponin I)
and tropomyosin subunits (tropomyosin-α chain
and tropomyosin-β chain) were retrieved from the
UniProt database (https://www.uniprot.org) (12). For
each species, five peptidic sequences were obtained
(Table 1).
Table 1. Data from the proteins retrieved from the UniProt database.
Organism Protein Amino acids length Entry code (ID)
Bos taurus
Troponin C 161 Q148C2|Q148C2_BOVIN
Troponin T 270 Q8MKI3|TNNT3_BOVIN
Troponin I 178 F6QIC1|F6QIC1_BOVIN
Tropomyosin-α 284 Q5KR49|TPM1_BOVIN
Tropomyosin-β 284 Q5KR48|TPM2_BOVIN
Gallus gallus
Troponin C 163 P02588|TNNC2_CHICK
Troponin T 263 P12620|TNNT3_CHICK
Troponin I 183 P68246|TNNI2_CHICK
Tropomyosin-α 284 P04268|TPM1_CHICK
Tropomyosin-β 284 P19352|TPM2_CHICK
Sus scrofa
Troponin C 159 P02587|TNNC2_PIG
Troponin T 271 Q75NG9|TNNT3_PIG
Troponin I 182 Q4JH15|Q4JH15_PIG
Tropomyosin-α 248 A0A4X1TTA0|A0A4X1TTA0_PIG
Tropomyosin-β 287 A1X899|A1X899_PIG
Bioactivity profiling
In silico enzymatic digestion and bioactivity profiling
were analyzed using the BIOPEP-UWM database (13)
especially on these derived from foods and being
constituents of diets that prevent development
of chronic diseases. The database is continuously
updated and modified. The addition of new
peptides and the introduction of new information
about the existing ones (e.g., chemical codes
and references to other databases) and served to
evaluate troponin and tropomyosin as a source of
4Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 30 | Number 01 | Article 347310Jorge Andrés Barrero, María Alejandra Barrero-Casallas, Angélica María González-Clavijo, Marcela Cruz-González
bioactive peptides. Gastrointestinal digestion was
simulated through in silico proteolysis catalyzed by
enzymes involved in gastric and pancreatic digestion
in the human digestive system; pepsin (E.C 3.4.23.1)
[pH<2], trypsin (E.C 3.4.21.4), and chymotrypsin
(E.C 3.4.21.1). The bioactive profile was assessed for
each polypeptide and later on compared between
species according to the parameters stated by
Minkiewicz et al. (13). The frequency of release of
bioactive fragments with given activity in a protein
sequence (A E) was determined using the equation 1:
E dA N
= [1]
where “d” is the number of peptides with specific
activity (e.g., DPP-IV inhibitors) and “N” is the
number of residues in the sequence.
Further on, the relative frequency percentage of
fragments release (W) was calculated as stated in
equation 2 based on a (total number of released
biopeptides with given activity) and the results
obtained from equation 1.
t
S t
a
A N
= [2]
At last, the bioactive profile of the whole set
of proteins derived from a single species was
compared. The mean occurrence of bioactive
fragments in a set of proteins (A S) was calculated
using equation 3:
t
S t
a
A N
= [3]
where “at” represents the total number of bioactive
peptides with a given activity, and “Nt” is the number
of amino acids in the whole set of proteins.
The parameter A S enabled a comparative analysis
of the bioactive potential of the proteins between
the species under study. Statistical analysis was
conducted to determine a significant difference in the
bioactive profile of troponin and tropomyosin derived
from each of the three species. A one-way ANOVA
test was applied with a significance level of 0.05.
Therapeutic potential assessment
Biopeptides obtained from meat digestion were
screened to evaluate the therapeutic properties
based on gastrointestinal absorption probability
(GI-absorption) determined through the SwissADME
web tool (14) and ADME/T lab platform (10). Those
peptides classified as GI-absorbable in both web
tools were subjected to pharmacokinetic profiling,
drug-likeness analysis, and target prediction. The
compliance with Lipinski’s rule of 5, Ghose, Veber,
Egan, and Muegge drug-likeness rules were assessed.
Absorption, distribution, metabolism, excretion, and
toxicity (ADME/T) parameters were predicted for the
GI-absorbable biopeptides intending to develop a
pharmacokinetic profile for each molecule. The target
proteins for these peptides were determined on the
SwissTarget-Prediction platform by searching for the
binding affinity for specific receptors/enzymes (11).
Statistical analyses were performed using a one-way
ANOVA test with a significance level of 0.05.
RESULTS AND DISCUSSION
Troponin and Tropomyosin bioactive profile
A simulated gastric and pancreatic digestion
was performed in silico, resulting in 40 bioactive
peptides released from all the myofibrillar proteins
isoforms analyzed. The bioactive profile of troponin
and tropomyosin subunits from each species
was assessed based on the biopeptide release
frequency. The relative frequency percentage of
released fragments (W) was determined (Figures 1
and 2). The bioactive profile of the set of proteins
from a single species was analyzed based on the A S
parameter (Figure 3).
The bioactive profile of troponin subunits, assessed
by the W parameters, revealed no significant
difference (p>0.05) between the isoforms of the
species under study (Figure 1). Seven classes of
bioactive peptides were reported: angiotensin-
converting enzyme (ACE) inhibitors, antioxidants,
stimulating, dipeptidyl-peptidase IV (DPP-IV)
inhibitors, calmodulin-dependent phosphodiesterase
(CaMPDE) inhibitors, renin inhibitors, and dipeptidyl-
peptidase III (DPP-III) inhibitors.
Two types of bioactive peptides were the most
frequently released upon enzymatic digestion: DPP-
IV inhibitors (DPP-IVi) and ACE inhibitors (ACEi). As
presented in Figure 1, troponin T isoforms showed
a release frequency of DPP-IVi >34.0%, with the
highest value attributed to pork (pig) (40.2%). The
release frequency of DPP-IVi for troponin I was
greater than 40.0%, and chicken reported the
highest frequency among all three species (63.2%).
At last, the troponin C release frequency of DPP-IVi
was higher than 24.0% for all three species, and beef
(cow) exhibited the highest score (36.8%).
bioactive peptides. Gastrointestinal digestion was
simulated through in silico proteolysis catalyzed by
enzymes involved in gastric and pancreatic digestion
in the human digestive system; pepsin (E.C 3.4.23.1)
[pH<2], trypsin (E.C 3.4.21.4), and chymotrypsin
(E.C 3.4.21.1). The bioactive profile was assessed for
each polypeptide and later on compared between
species according to the parameters stated by
Minkiewicz et al. (13). The frequency of release of
bioactive fragments with given activity in a protein
sequence (A E) was determined using the equation 1:
E dA N
= [1]
where “d” is the number of peptides with specific
activity (e.g., DPP-IV inhibitors) and “N” is the
number of residues in the sequence.
Further on, the relative frequency percentage of
fragments release (W) was calculated as stated in
equation 2 based on a (total number of released
biopeptides with given activity) and the results
obtained from equation 1.
t
S t
a
A N
= [2]
At last, the bioactive profile of the whole set
of proteins derived from a single species was
compared. The mean occurrence of bioactive
fragments in a set of proteins (A S) was calculated
using equation 3:
t
S t
a
A N
= [3]
where “at” represents the total number of bioactive
peptides with a given activity, and “Nt” is the number
of amino acids in the whole set of proteins.
The parameter A S enabled a comparative analysis
of the bioactive potential of the proteins between
the species under study. Statistical analysis was
conducted to determine a significant difference in the
bioactive profile of troponin and tropomyosin derived
from each of the three species. A one-way ANOVA
test was applied with a significance level of 0.05.
Therapeutic potential assessment
Biopeptides obtained from meat digestion were
screened to evaluate the therapeutic properties
based on gastrointestinal absorption probability
(GI-absorption) determined through the SwissADME
web tool (14) and ADME/T lab platform (10). Those
peptides classified as GI-absorbable in both web
tools were subjected to pharmacokinetic profiling,
drug-likeness analysis, and target prediction. The
compliance with Lipinski’s rule of 5, Ghose, Veber,
Egan, and Muegge drug-likeness rules were assessed.
Absorption, distribution, metabolism, excretion, and
toxicity (ADME/T) parameters were predicted for the
GI-absorbable biopeptides intending to develop a
pharmacokinetic profile for each molecule. The target
proteins for these peptides were determined on the
SwissTarget-Prediction platform by searching for the
binding affinity for specific receptors/enzymes (11).
Statistical analyses were performed using a one-way
ANOVA test with a significance level of 0.05.
RESULTS AND DISCUSSION
Troponin and Tropomyosin bioactive profile
A simulated gastric and pancreatic digestion
was performed in silico, resulting in 40 bioactive
peptides released from all the myofibrillar proteins
isoforms analyzed. The bioactive profile of troponin
and tropomyosin subunits from each species
was assessed based on the biopeptide release
frequency. The relative frequency percentage of
released fragments (W) was determined (Figures 1
and 2). The bioactive profile of the set of proteins
from a single species was analyzed based on the A S
parameter (Figure 3).
The bioactive profile of troponin subunits, assessed
by the W parameters, revealed no significant
difference (p>0.05) between the isoforms of the
species under study (Figure 1). Seven classes of
bioactive peptides were reported: angiotensin-
converting enzyme (ACE) inhibitors, antioxidants,
stimulating, dipeptidyl-peptidase IV (DPP-IV)
inhibitors, calmodulin-dependent phosphodiesterase
(CaMPDE) inhibitors, renin inhibitors, and dipeptidyl-
peptidase III (DPP-III) inhibitors.
Two types of bioactive peptides were the most
frequently released upon enzymatic digestion: DPP-
IV inhibitors (DPP-IVi) and ACE inhibitors (ACEi). As
presented in Figure 1, troponin T isoforms showed
a release frequency of DPP-IVi >34.0%, with the
highest value attributed to pork (pig) (40.2%). The
release frequency of DPP-IVi for troponin I was
greater than 40.0%, and chicken reported the
highest frequency among all three species (63.2%).
At last, the troponin C release frequency of DPP-IVi
was higher than 24.0% for all three species, and beef
(cow) exhibited the highest score (36.8%).
5Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
DPP-IV is a serine protease involved in the
degradation of incretin hormones, glucagon-
like peptide 1 (GLP-1) and glucose-dependent
insulinotropic peptide (GIP) (15). These hormones
exert diverse physiological functions, including
insulin secretion potentiation following food intake,
slowing gastric emptying, and satiety stimulation
(15). Thus, DPP-IV inhibition is a suitable therapeutic
approach for glycemic regulation in hyperglycemic
disorders. The high release frequency values of
DPP-IVi bioactive peptides suggest that troponin
and tropomyosin dietary intake hold therapeutic
proper ties for glycemia reduction. Although
these outcomes are mainly theoretical, in vivo
experimentation supports the results of this study,
as stated by Casanova-Martí et al. (16), who found
that biopeptides derived from chicken had a high
DPP-IV inhibitory capacity and improved GLP-1
secretion and glucose tolerance in rats.
In addition to DPP-IVi, ACE inhibitors were the
second most frequently obtained bioactive peptides
(Figure 1). Among the products of troponin C
digestion, ACEi was the most commonly released
(>54.0%); and chicken isoforms showed the highest
W value from all three species (58.1%). The frequency
of ACEi biopeptides release from troponin T was
above 32.0%, and similar to troponin C, the chicken
isoform scored the highest (35.5%). On the contrary,
W parameter analysis of troponin I hydrolysis
showed that the chicken isoform had the lowest
ACEi release frequency (31.0%), while the other two
were above 42.0%.
ACE is a carboxypeptidase that catalyzes the
activation of angiotensin II. It is one of the main
hormones of the renin-angiotensin-aldosterone
system involved in fluid retention by aldosterone
secretion stimulation and blood pressure increase
by induced vascular spasm (17). ACE inhibition has
long been known for its favorable outcomes in
hypertension therapy. Bioactive peptides such as
Val-Tyr and Ile-Trp have shown vascular resistance
modulation (18). However, several biopeptides lack in
vitro evidence of ACE inhibition and blood pressure
reduction. Thus, in vivo screening studies are still
required to determine suitable anti-hypertensive
properties when orally delivered.
Antioxidant biopeptides were obtained from the
troponin isoforms of all three species (Figure 1).
Results varied irregularly. Troponin T isoforms
showed a release frequency (W) greater than 10.0%
and pork (pig) exhibited the highest value (12.00%).
Troponin C revealed values greater than 8.0%, with
the chicken isoform exhibiting more than twice the
Figure 1. Relative frequency percentage of bioactive peptides released from troponin. (A) Bioactive profile of troponin T. (B) Bioactive
profile of troponin C. (C) Bioactive profile of troponin I.
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
DPP-IV is a serine protease involved in the
degradation of incretin hormones, glucagon-
like peptide 1 (GLP-1) and glucose-dependent
insulinotropic peptide (GIP) (15). These hormones
exert diverse physiological functions, including
insulin secretion potentiation following food intake,
slowing gastric emptying, and satiety stimulation
(15). Thus, DPP-IV inhibition is a suitable therapeutic
approach for glycemic regulation in hyperglycemic
disorders. The high release frequency values of
DPP-IVi bioactive peptides suggest that troponin
and tropomyosin dietary intake hold therapeutic
proper ties for glycemia reduction. Although
these outcomes are mainly theoretical, in vivo
experimentation supports the results of this study,
as stated by Casanova-Martí et al. (16), who found
that biopeptides derived from chicken had a high
DPP-IV inhibitory capacity and improved GLP-1
secretion and glucose tolerance in rats.
In addition to DPP-IVi, ACE inhibitors were the
second most frequently obtained bioactive peptides
(Figure 1). Among the products of troponin C
digestion, ACEi was the most commonly released
(>54.0%); and chicken isoforms showed the highest
W value from all three species (58.1%). The frequency
of ACEi biopeptides release from troponin T was
above 32.0%, and similar to troponin C, the chicken
isoform scored the highest (35.5%). On the contrary,
W parameter analysis of troponin I hydrolysis
showed that the chicken isoform had the lowest
ACEi release frequency (31.0%), while the other two
were above 42.0%.
ACE is a carboxypeptidase that catalyzes the
activation of angiotensin II. It is one of the main
hormones of the renin-angiotensin-aldosterone
system involved in fluid retention by aldosterone
secretion stimulation and blood pressure increase
by induced vascular spasm (17). ACE inhibition has
long been known for its favorable outcomes in
hypertension therapy. Bioactive peptides such as
Val-Tyr and Ile-Trp have shown vascular resistance
modulation (18). However, several biopeptides lack in
vitro evidence of ACE inhibition and blood pressure
reduction. Thus, in vivo screening studies are still
required to determine suitable anti-hypertensive
properties when orally delivered.
Antioxidant biopeptides were obtained from the
troponin isoforms of all three species (Figure 1).
Results varied irregularly. Troponin T isoforms
showed a release frequency (W) greater than 10.0%
and pork (pig) exhibited the highest value (12.00%).
Troponin C revealed values greater than 8.0%, with
the chicken isoform exhibiting more than twice the
Figure 1. Relative frequency percentage of bioactive peptides released from troponin. (A) Bioactive profile of troponin T. (B) Bioactive
profile of troponin C. (C) Bioactive profile of troponin I.
6Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 30 | Number 01 | Article 347310Jorge Andrés Barrero, María Alejandra Barrero-Casallas, Angélica María González-Clavijo, Marcela Cruz-González
value of the two other isoforms (16.2%). Finally,
troponin I presented a release frequency greater
than 5.0%, with beef (cow) scoring the highest
value (7.6%). While antioxidant bioactive peptides
released from meat digestion hold diverse functions,
the peptides obtained act mainly by free radicals
scavenging and oxidative stress reduction (19, 20).
Moreover, DPP-III inhibitors (DPP-IIIi) were reported
as hydrolysis products of troponin T exclusively. The
release frequency (W) of this biopeptide was above
4.0% for all three species, and beef (cow) showed
the highest score (7.3%). Results suggest a dietary
intake of DPP-IIIi biopeptides. However, while DPP-
III inhibition has shown improved hemodynamics in
rodents (21), the role of DPP-III bioactive peptides as
hemodynamic enhancers awaits to be unrevealed.
Troponin T was the only source of stimulating
b i o p e p t i d e s , c a l m o d u l i n - d e p e n d e n t
phosphodiesterase (CaMPDE) inhibitors, and renin
blockers. For these three classes of biopeptides, the
release frequency (W) was higher than 4.0%, with the
chicken isoform slightly above the other two (5.3%)
(Figure 1). Stimulating peptides enhance glucose
uptake (22); however, its mechanism is not entirely
understood. CaMPDE is a phosphodiesterase
involved in the cross-talk between cAMP and Ca 2+
signaling in a wide range of cell populations (23).
CaMPDE inhibitors have been shown to exert anti-
inflammatory effects on glial cells and could be
therapeutic in neurodegenerative disorders (24).
Renin-inhibitors appear to block renin’s cleavage
from angiotensinogen to angiotensin I. Despite
the poor outcomes of renin-inhibiting drugs for
hypertension therapy (25), bioactive peptides
derived from meat digestion could interact
synergically with commonly administered drugs
for hypertension therapy.
Similar to the bioactive profile of troponin subunits,
no significant difference was found in biopeptides
release frequency (p>0.05) between the tropomyosin
isoforms (Figure 2). Tropomyosin-α exhibits the
highest number of bioactive peptides released upon
gastric and pancreatic digestion, and except for pig’s
isoform, the most frequently released biopeptides
were DPP-IV inhibitors.
Tropomyosin-β showed a high bioactive profile
for DPP-IVi biopeptides release; however, only
three classes of bioactive peptides were obtained
from its digestion (Figure 2). For DPP-IVi, a release
frequency >50.0% was reported, and beef (cow)
seems to be the isoform that is a better precursor of
these peptides. ACEi release frequency was highest
for the chicken isoform (36.7%), and antioxidant
biopeptides were more frequently released from
the pig’s isoform (17.9%).
Figure 2. Relative frequency percentage of bioactive peptides
release from tropomyosin. (A) Bioactive profile of tropomyosin-β.
(B) Bioactive profile of tropomyosin-α.
On the other hand, tropomyosin-α reported being
a rich source of ACEi biopeptides with a release
frequency greater than 33.0% and with pig’s isoform
scoring the highest value (35.2%), The release
frequency of DPP-IVi biopeptides was >50.0% for
beef (cow) and chicken, and <30.0% for pork (pig).
On the other hand, antioxidant bioactive peptides
reported a release frequency of over 10.0%, and
pig’s isoform exhibited the highest value (14.8%).
Stimulating peptides, renin inhibitors, CaMPDE
inhibitors, and regulating biopeptides all showed a
release frequency lower than 7.0%.
Most of the biological activities repor ted in
tropomyosin digestion end-products were also
identified in dipeptides released from troponin
digestion, except for the regulating biopeptides.
Dipeptide with sequence Asp-Tyr was the only
regulating biopeptide obtained purely from beef
(cow) tropomyosin-α. This bioactive peptide exhibits
a regulation of ionic flux across the cell membrane,
showing electrophysiological modulation on
neurons in rodents (26).
value of the two other isoforms (16.2%). Finally,
troponin I presented a release frequency greater
than 5.0%, with beef (cow) scoring the highest
value (7.6%). While antioxidant bioactive peptides
released from meat digestion hold diverse functions,
the peptides obtained act mainly by free radicals
scavenging and oxidative stress reduction (19, 20).
Moreover, DPP-III inhibitors (DPP-IIIi) were reported
as hydrolysis products of troponin T exclusively. The
release frequency (W) of this biopeptide was above
4.0% for all three species, and beef (cow) showed
the highest score (7.3%). Results suggest a dietary
intake of DPP-IIIi biopeptides. However, while DPP-
III inhibition has shown improved hemodynamics in
rodents (21), the role of DPP-III bioactive peptides as
hemodynamic enhancers awaits to be unrevealed.
Troponin T was the only source of stimulating
b i o p e p t i d e s , c a l m o d u l i n - d e p e n d e n t
phosphodiesterase (CaMPDE) inhibitors, and renin
blockers. For these three classes of biopeptides, the
release frequency (W) was higher than 4.0%, with the
chicken isoform slightly above the other two (5.3%)
(Figure 1). Stimulating peptides enhance glucose
uptake (22); however, its mechanism is not entirely
understood. CaMPDE is a phosphodiesterase
involved in the cross-talk between cAMP and Ca 2+
signaling in a wide range of cell populations (23).
CaMPDE inhibitors have been shown to exert anti-
inflammatory effects on glial cells and could be
therapeutic in neurodegenerative disorders (24).
Renin-inhibitors appear to block renin’s cleavage
from angiotensinogen to angiotensin I. Despite
the poor outcomes of renin-inhibiting drugs for
hypertension therapy (25), bioactive peptides
derived from meat digestion could interact
synergically with commonly administered drugs
for hypertension therapy.
Similar to the bioactive profile of troponin subunits,
no significant difference was found in biopeptides
release frequency (p>0.05) between the tropomyosin
isoforms (Figure 2). Tropomyosin-α exhibits the
highest number of bioactive peptides released upon
gastric and pancreatic digestion, and except for pig’s
isoform, the most frequently released biopeptides
were DPP-IV inhibitors.
Tropomyosin-β showed a high bioactive profile
for DPP-IVi biopeptides release; however, only
three classes of bioactive peptides were obtained
from its digestion (Figure 2). For DPP-IVi, a release
frequency >50.0% was reported, and beef (cow)
seems to be the isoform that is a better precursor of
these peptides. ACEi release frequency was highest
for the chicken isoform (36.7%), and antioxidant
biopeptides were more frequently released from
the pig’s isoform (17.9%).
Figure 2. Relative frequency percentage of bioactive peptides
release from tropomyosin. (A) Bioactive profile of tropomyosin-β.
(B) Bioactive profile of tropomyosin-α.
On the other hand, tropomyosin-α reported being
a rich source of ACEi biopeptides with a release
frequency greater than 33.0% and with pig’s isoform
scoring the highest value (35.2%), The release
frequency of DPP-IVi biopeptides was >50.0% for
beef (cow) and chicken, and <30.0% for pork (pig).
On the other hand, antioxidant bioactive peptides
reported a release frequency of over 10.0%, and
pig’s isoform exhibited the highest value (14.8%).
Stimulating peptides, renin inhibitors, CaMPDE
inhibitors, and regulating biopeptides all showed a
release frequency lower than 7.0%.
Most of the biological activities repor ted in
tropomyosin digestion end-products were also
identified in dipeptides released from troponin
digestion, except for the regulating biopeptides.
Dipeptide with sequence Asp-Tyr was the only
regulating biopeptide obtained purely from beef
(cow) tropomyosin-α. This bioactive peptide exhibits
a regulation of ionic flux across the cell membrane,
showing electrophysiological modulation on
neurons in rodents (26).
7Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
The bioactive profile of the myofibrillar proteins
from the three species revealed no significant
difference (p>0.05) in terms of the mean frequency
(AS) of bioactive peptides occurrence (Figure 3).
Nonetheless, variations were observed regarding
the biological activity of the biopeptides obtained.
DPP-IVi peptides were most frequently released
from chicken (G. gallus) and pig (S. scrofa) troponin
and tropomyosin. The findings of Martini et al. (27)
support the results of this study, as they found out
by in vitro experimentation that pork had a higher
DPP-IV inhibiting activity than chicken meat, yet all
the types of meat analyzed (pork, beef, chicken,
and turkey) were determined to be good sources
of DPP-IVi bioactive peptides. On the other hand,
troponin and tropomyosin isoforms from the cow
exhibited the highest release frequency of ACEi
biopeptides from all three species. Similarly, Mora
et al. (28) analyzed meat proteins and concluded
that meat is a rich source of peptides with ACE
inhibitory properties.
Antioxidant, stimulating, renin-inhibiting, CaMPDE-
inhibiting, and DPP-III-inhibiting bioactive peptides
reported a low mean frequency (A S ) of bioactive
peptides occurrence when compared to DPP-IVi and
ACEi peptides. Regulating peptides, on the other
hand, were only obtained from beef. Compared to
DPP-IV and ACE inhibition, these functions have
been less studied and still need to be revealed.
Bioactivity and drug-like properties of GI-
absorbable biopeptides
The biopeptides obtained from gastric and
pancreatic simulated digestion were screened based
on the classification as gastrointestinal-absorbable
(GI-absorbable) biopeptides in SwissADME
and ADME/Tlab platforms. The bioactivity and
half-maximal effective concentration for DPP-IV
inhibition (EC50) of GI-absorbable peptides are
shown in Table 2.
Out of 40 bioactive peptides released from
enzymatic proteolysis of troponin and tropomyosin
isoforms, 7 were predicted to be easily absorbed
by enterocytes. Likely absorbable dipeptides were
obtained from all three species’ isoforms except for
three peptides. Ale-Phe and Cys-Phe were obtained
only from G. gallus troponin C, and Gly-Leu was
exclusively released from S. scrofa tropomyosin-β
digestion. Hence, among the species under study,
G. gallus proteins appear to be a better source
of absorbable bioactive peptides. Ryan et al. (34)
showed that chicken breast muscle is a rich source of
several bioactive peptides; however, the evaluation
of bioactive fragments release was performed with
enzymes foreign to the human digestive system.
Figure 3. Mean frequency of occurrence of bioactive peptides in troponin and tropomyosin from beef (B. taurus), chicken (G. gallus)
and pork (S. scrofa) fast skeletal muscle.
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
The bioactive profile of the myofibrillar proteins
from the three species revealed no significant
difference (p>0.05) in terms of the mean frequency
(AS) of bioactive peptides occurrence (Figure 3).
Nonetheless, variations were observed regarding
the biological activity of the biopeptides obtained.
DPP-IVi peptides were most frequently released
from chicken (G. gallus) and pig (S. scrofa) troponin
and tropomyosin. The findings of Martini et al. (27)
support the results of this study, as they found out
by in vitro experimentation that pork had a higher
DPP-IV inhibiting activity than chicken meat, yet all
the types of meat analyzed (pork, beef, chicken,
and turkey) were determined to be good sources
of DPP-IVi bioactive peptides. On the other hand,
troponin and tropomyosin isoforms from the cow
exhibited the highest release frequency of ACEi
biopeptides from all three species. Similarly, Mora
et al. (28) analyzed meat proteins and concluded
that meat is a rich source of peptides with ACE
inhibitory properties.
Antioxidant, stimulating, renin-inhibiting, CaMPDE-
inhibiting, and DPP-III-inhibiting bioactive peptides
reported a low mean frequency (A S ) of bioactive
peptides occurrence when compared to DPP-IVi and
ACEi peptides. Regulating peptides, on the other
hand, were only obtained from beef. Compared to
DPP-IV and ACE inhibition, these functions have
been less studied and still need to be revealed.
Bioactivity and drug-like properties of GI-
absorbable biopeptides
The biopeptides obtained from gastric and
pancreatic simulated digestion were screened based
on the classification as gastrointestinal-absorbable
(GI-absorbable) biopeptides in SwissADME
and ADME/Tlab platforms. The bioactivity and
half-maximal effective concentration for DPP-IV
inhibition (EC50) of GI-absorbable peptides are
shown in Table 2.
Out of 40 bioactive peptides released from
enzymatic proteolysis of troponin and tropomyosin
isoforms, 7 were predicted to be easily absorbed
by enterocytes. Likely absorbable dipeptides were
obtained from all three species’ isoforms except for
three peptides. Ale-Phe and Cys-Phe were obtained
only from G. gallus troponin C, and Gly-Leu was
exclusively released from S. scrofa tropomyosin-β
digestion. Hence, among the species under study,
G. gallus proteins appear to be a better source
of absorbable bioactive peptides. Ryan et al. (34)
showed that chicken breast muscle is a rich source of
several bioactive peptides; however, the evaluation
of bioactive fragments release was performed with
enzymes foreign to the human digestive system.
Figure 3. Mean frequency of occurrence of bioactive peptides in troponin and tropomyosin from beef (B. taurus), chicken (G. gallus)
and pork (S. scrofa) fast skeletal muscle.
8Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 30 | Number 01 | Article 347310Jorge Andrés Barrero, María Alejandra Barrero-Casallas, Angélica María González-Clavijo, Marcela Cruz-González
Two main properties were identified from the
bioactive profile of the GI-absorbable biopeptides
released: anti-diabetic and anti-hypertensive.
DPP-IV inhibitors and glucose uptake stimulating
biopeptides are highlighted by previously exposed
mechanisms as potential glycemic-regulating
agents in type 2 diabetes mellitus therapy. These
biopeptides could prolong insulin secretion by
similar mechanisms to DPP-IV inhibiting drugs such as
sitagliptin. Likewise, ACE inhibitors reduce peripheral
vascular resistance by downregulating angiotensin II
production and bradykinin degradation, therefore
sharing similar action mechanisms as ACE-inhibiting
drugs. The biological properties of troponin and
tropomyosin hydrolysates reported in this study
suggest that biopeptides might hold potential
health-promoting properties in type 2 diabetes
mellitus and hypertension therapy. Nonetheless,
determining these nutraceuticals’ therapeutic
effects is subject to future studies.
The half-maximal concentration of DPP-IV inhibition
(EC50) values for the GI-absorbable biopeptides
were retrieved to assess the therapeutic potency
of the GI-absorbable biopeptides. As shown in
Table 2, the therapeutic potency varied significantly
among the released peptides. In vivo studies
demonstrated enhanced insulin sensitivity and
postprandial glycemic reduction in diabetic animal
models when DPP-IVi peptides were administered
(8). Nonetheless, when compared to approved
drugs, DDP-IVi biopeptides exhibit a much higher
EC50 than DPP-IVi drugs (e.g., sitagliptin [<0.03μM]
(35)). Similarly, ACEi peptides show higher EC50
values than ACEi drugs (e.g., captopril [<0.1μM]
(36)). These results indicated that biopeptides
held a lower therapeutic potency than formerly
approved drugs and were not suited to replace
current pharmacotherapy. A lower half-life, higher
clearance, and lesser affinity for the target protein
might be some of the factors underlying the lower
therapeutic potency of nutraceuticals. Still, future
studies might reveal if biopeptides, concomitantly
administered to drugs, could enhance their potency.
Currently, evidence in humans is still unknown,
partly due to the scarcity of reports regarding the
pharmacokinetics of bioactive peptides. Aiming
to elucidate this query, drug-likeness compliance
and the pharmacokinetic profile of GI-absorbable
biopeptides was assessed, and results are presented
in Table 3.
Table 2. Bioactivity of meat-derived GI-absorbable bioactive peptides.
Biopeptide Sequence Action(s) Source EC50 (μM) Reference
Val-Leu (1) Glucose uptake stimulating
(2) DPP-IV inhibitor
B. taurus: Tm-α, TnT
G. gallus: Tm-α, TnT
S. scrofa: Tm-α, TnT
(1): -
(2): 74.00 (22, 29)
Ala-Leu DPP-IV inhibitor
B. taurus: Tm-α, Tm-β, TnT
G. gallus: Tm-α, Tm-β, TnT, TnI
S. scrofa: TnT, TnI
882.1 (30)
Pro-Leu ACE inhibitor
B. taurus: TnT
G. gallus: TnT
S. scrofa: TnT
337.3 (31)
Ile-Phe ACE inhibitor
B. taurus: TnC
G. gallus: TnC
S. scrofa: TnC
930.0 (32)
Ala-Phe ACE inhibitor G. gallus: TnC 190.0 (32)
Cys-Phe ACE inhibitor G. gallus: TnC 2.0 (33)
Gly-Leu (1) ACE inhibitor
(2) DPP-IV inhibitor S. scrofa: Tm-β (1): 2500.0
(2): 2615.0 (30, 32)
DPP-IV, Dipeptidyl-Peptidase IV; ACE, Angiotensin Converting Enzyme; Tm-α, Tropomyosin-α; Tm-β, Tropomyosin-β; TnT, Troponin T; TnC, Troponin C; TnI, Troponin
I; EC50, Half maximal effective concentration.
Two main properties were identified from the
bioactive profile of the GI-absorbable biopeptides
released: anti-diabetic and anti-hypertensive.
DPP-IV inhibitors and glucose uptake stimulating
biopeptides are highlighted by previously exposed
mechanisms as potential glycemic-regulating
agents in type 2 diabetes mellitus therapy. These
biopeptides could prolong insulin secretion by
similar mechanisms to DPP-IV inhibiting drugs such as
sitagliptin. Likewise, ACE inhibitors reduce peripheral
vascular resistance by downregulating angiotensin II
production and bradykinin degradation, therefore
sharing similar action mechanisms as ACE-inhibiting
drugs. The biological properties of troponin and
tropomyosin hydrolysates reported in this study
suggest that biopeptides might hold potential
health-promoting properties in type 2 diabetes
mellitus and hypertension therapy. Nonetheless,
determining these nutraceuticals’ therapeutic
effects is subject to future studies.
The half-maximal concentration of DPP-IV inhibition
(EC50) values for the GI-absorbable biopeptides
were retrieved to assess the therapeutic potency
of the GI-absorbable biopeptides. As shown in
Table 2, the therapeutic potency varied significantly
among the released peptides. In vivo studies
demonstrated enhanced insulin sensitivity and
postprandial glycemic reduction in diabetic animal
models when DPP-IVi peptides were administered
(8). Nonetheless, when compared to approved
drugs, DDP-IVi biopeptides exhibit a much higher
EC50 than DPP-IVi drugs (e.g., sitagliptin [<0.03μM]
(35)). Similarly, ACEi peptides show higher EC50
values than ACEi drugs (e.g., captopril [<0.1μM]
(36)). These results indicated that biopeptides
held a lower therapeutic potency than formerly
approved drugs and were not suited to replace
current pharmacotherapy. A lower half-life, higher
clearance, and lesser affinity for the target protein
might be some of the factors underlying the lower
therapeutic potency of nutraceuticals. Still, future
studies might reveal if biopeptides, concomitantly
administered to drugs, could enhance their potency.
Currently, evidence in humans is still unknown,
partly due to the scarcity of reports regarding the
pharmacokinetics of bioactive peptides. Aiming
to elucidate this query, drug-likeness compliance
and the pharmacokinetic profile of GI-absorbable
biopeptides was assessed, and results are presented
in Table 3.
Table 2. Bioactivity of meat-derived GI-absorbable bioactive peptides.
Biopeptide Sequence Action(s) Source EC50 (μM) Reference
Val-Leu (1) Glucose uptake stimulating
(2) DPP-IV inhibitor
B. taurus: Tm-α, TnT
G. gallus: Tm-α, TnT
S. scrofa: Tm-α, TnT
(1): -
(2): 74.00 (22, 29)
Ala-Leu DPP-IV inhibitor
B. taurus: Tm-α, Tm-β, TnT
G. gallus: Tm-α, Tm-β, TnT, TnI
S. scrofa: TnT, TnI
882.1 (30)
Pro-Leu ACE inhibitor
B. taurus: TnT
G. gallus: TnT
S. scrofa: TnT
337.3 (31)
Ile-Phe ACE inhibitor
B. taurus: TnC
G. gallus: TnC
S. scrofa: TnC
930.0 (32)
Ala-Phe ACE inhibitor G. gallus: TnC 190.0 (32)
Cys-Phe ACE inhibitor G. gallus: TnC 2.0 (33)
Gly-Leu (1) ACE inhibitor
(2) DPP-IV inhibitor S. scrofa: Tm-β (1): 2500.0
(2): 2615.0 (30, 32)
DPP-IV, Dipeptidyl-Peptidase IV; ACE, Angiotensin Converting Enzyme; Tm-α, Tropomyosin-α; Tm-β, Tropomyosin-β; TnT, Troponin T; TnC, Troponin C; TnI, Troponin
I; EC50, Half maximal effective concentration.
9Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
Drug-likeness guidelines state several parameters
to evaluate the compliance of compounds with
physicochemical proper ties crucial for drug
candidates. DPP-IVi bioactive peptides were
assessed based on five drug-likeness rules (Lipinski’s
rule of 5, Ghose filter, Veber’s rules, Egan filter, and
Muegge filter) which evaluated: molecular weight,
molar refractivity, lipophilicity (LogP), topological
polar surface area (TPSA), the number of rings,
carbons, rotatable bonds, heteroatoms, hydrogen-
bond donors and hydrogen-bond acceptors (14).
As shown in Table 3, lipophilicity (LogP) was the
only violated criterion by all dipeptides except
Ile-Phe. The violation of the LogP parameter could
be attributed to the amphoteric properties of
amino acids. In concordance, Doroshuck et al. (37)
identified that zwitterions made up of L-amino acids
tend to exhibit low LogP values establishing them
as highly hydrophilic compounds. Nevertheless,
regardless of lipophilicity’s slight violation of drug-
likeness rules, the lower hydrophobicity could
enhance gastrointestinal absorption, bioavailability,
and ligand-target interaction (38, 39). While in silico
evaluation of drug-like properties is limited by the
theoretical approach based on topological and
molecular descriptors, these parameters determine
important chemical features that are expected for
a drug candidate when tested in vivo (40). Hence,
based on the results of this evaluation, bioactive
peptides are predicted to exhibit proper drug-like
properties when orally administered.
ADME/T analysis was carried out to determine
the pharmacokinetics (absorption, distribution,
metabolism, excretion, and toxicity) of bioactive
peptides. As previously mentioned, seven peptides
were filtered according to absorption prediction,
and bioavailability was assessed. F(≥30%) value
Table 3. Biopeptides drug-likeness compliance and pharmacokinetic profile.
Parameter Bioactive peptide
Val-Leu Ala-Leu Pro-Leu Ile-Phe Ala-Phe Cys-Phe Gly-Leu
Drug-likeness compliance (%)
Lipinski 100.0 100.0 100.0 100.0 100.0 100.00 100.0
Ghose 100.0 100.0 100.0 100.0 100.0 100. 75.0
(LogP<-0.4) a
Veber 100.0 100.0 100.0 100.0 100.0 100.0 100.0
Egan 100.0 100.0 100.0 100.0 100.0 100.0 100.0
Muegge 88.9
(LogP<-2)a
88.9
(LogP<-2)a
88.9
(LogP<-2)a 100.0 88.9
(LogP<-2) a
88.9
(LogP<-2)a
88.9
(LogP<-2)a
Absorption
HIA (≥30%) High High High High High High High
F (≥30%) + + - + + - +
Distribution
PPB (%) 47.4 36.6 46.6 63.9 55.6 53.6 26.8
BBB +++ +++ ++ +++ +++ +++ +++
Metabolism
CYP450 interactions None None None None None None None
Excretion
T1/2 (h) 1.4 1.2 0.9 0.7 0.7 0.6 1.3
Clearance (cm3 ·min-1 ·kg-1 ) 1.3 1.2 1.4 1.5 1.5 1.4 1.0
Toxicity
DILI --- --- --- --- --- --- ---
HIA, Human Intestinal Absorption; F(≥30%), ≥30% Bioavailability; PPB, Plasma-protein binding; BBB, Blood brain barrier; CYP450, Cytochrome P450; T1/2, half-life; DILI,
Drug-Induced Liver Injury. aViolated criteria. (+): probability 0.5-0.7, (++); probability 0.7-0.9, (+++): probability 0.9-1.0, (-): probability 0.3-0.5, (---): probability 0.0-0.1.
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
Drug-likeness guidelines state several parameters
to evaluate the compliance of compounds with
physicochemical proper ties crucial for drug
candidates. DPP-IVi bioactive peptides were
assessed based on five drug-likeness rules (Lipinski’s
rule of 5, Ghose filter, Veber’s rules, Egan filter, and
Muegge filter) which evaluated: molecular weight,
molar refractivity, lipophilicity (LogP), topological
polar surface area (TPSA), the number of rings,
carbons, rotatable bonds, heteroatoms, hydrogen-
bond donors and hydrogen-bond acceptors (14).
As shown in Table 3, lipophilicity (LogP) was the
only violated criterion by all dipeptides except
Ile-Phe. The violation of the LogP parameter could
be attributed to the amphoteric properties of
amino acids. In concordance, Doroshuck et al. (37)
identified that zwitterions made up of L-amino acids
tend to exhibit low LogP values establishing them
as highly hydrophilic compounds. Nevertheless,
regardless of lipophilicity’s slight violation of drug-
likeness rules, the lower hydrophobicity could
enhance gastrointestinal absorption, bioavailability,
and ligand-target interaction (38, 39). While in silico
evaluation of drug-like properties is limited by the
theoretical approach based on topological and
molecular descriptors, these parameters determine
important chemical features that are expected for
a drug candidate when tested in vivo (40). Hence,
based on the results of this evaluation, bioactive
peptides are predicted to exhibit proper drug-like
properties when orally administered.
ADME/T analysis was carried out to determine
the pharmacokinetics (absorption, distribution,
metabolism, excretion, and toxicity) of bioactive
peptides. As previously mentioned, seven peptides
were filtered according to absorption prediction,
and bioavailability was assessed. F(≥30%) value
Table 3. Biopeptides drug-likeness compliance and pharmacokinetic profile.
Parameter Bioactive peptide
Val-Leu Ala-Leu Pro-Leu Ile-Phe Ala-Phe Cys-Phe Gly-Leu
Drug-likeness compliance (%)
Lipinski 100.0 100.0 100.0 100.0 100.0 100.00 100.0
Ghose 100.0 100.0 100.0 100.0 100.0 100. 75.0
(LogP<-0.4) a
Veber 100.0 100.0 100.0 100.0 100.0 100.0 100.0
Egan 100.0 100.0 100.0 100.0 100.0 100.0 100.0
Muegge 88.9
(LogP<-2)a
88.9
(LogP<-2)a
88.9
(LogP<-2)a 100.0 88.9
(LogP<-2) a
88.9
(LogP<-2)a
88.9
(LogP<-2)a
Absorption
HIA (≥30%) High High High High High High High
F (≥30%) + + - + + - +
Distribution
PPB (%) 47.4 36.6 46.6 63.9 55.6 53.6 26.8
BBB +++ +++ ++ +++ +++ +++ +++
Metabolism
CYP450 interactions None None None None None None None
Excretion
T1/2 (h) 1.4 1.2 0.9 0.7 0.7 0.6 1.3
Clearance (cm3 ·min-1 ·kg-1 ) 1.3 1.2 1.4 1.5 1.5 1.4 1.0
Toxicity
DILI --- --- --- --- --- --- ---
HIA, Human Intestinal Absorption; F(≥30%), ≥30% Bioavailability; PPB, Plasma-protein binding; BBB, Blood brain barrier; CYP450, Cytochrome P450; T1/2, half-life; DILI,
Drug-Induced Liver Injury. aViolated criteria. (+): probability 0.5-0.7, (++); probability 0.7-0.9, (+++): probability 0.9-1.0, (-): probability 0.3-0.5, (---): probability 0.0-0.1.
10Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 30 | Number 01 | Article 347310Jorge Andrés Barrero, María Alejandra Barrero-Casallas, Angélica María González-Clavijo, Marcela Cruz-González
predicts the probability of achieving a bioavailability
greater than 30% when orally administered (41).
Five of seven peptides scored >50% chance of
surpassing this threshold. Distribution evaluation
revealed that GI-absorbable bioactive peptides are
likely to cross the blood-brain barrier. Based on the
plasma protein binding probability results, peptides
are expected to remain in the bloodstream without
binding to carrying proteins.
Metabolism was assessed based on the interactions
with cytochrome enzymes (CYP450). This large family
of oxidoreductase enzymes are tightly involved in
xenobiotics metabolism; hence, it is important to
predict their interactions with drug-like compounds
(42). Bioactive peptides reported no single substrate-
like or inhibitory interaction with any CYP450. On the
other hand, when assessing toxicity, results show that
peptides induce no liver injury based on the DILI
parameter. Our findings support the hypothesis of
Acquah et al. (8) since in silico evaluation indicated
that biopeptides exert little complications in
humans in terms of toxicity and organ damage. At
last, excretion was evaluated based on half-life and
clearance rate. Half-life varied irregularly among
these peptides, and only three reported a value
higher than 1 hour. On the other hand, clearance
predicted values were all above 1 cm 3 ·min -1
·kg -1
.
These two parameters must be carefully considered,
as low half-life and high clearance could imply
an increased dosing regimen and consumption
frequency to achieve an effective therapeutic effect
(43). This work develops a non-previously analyzed
pharmacokinetic profile of bioactive peptides, yet,
in vivo studies are still imperative to conclude with
a greater degree of certainty.
Target Prediction
Bioactive peptides exhibit a doubtless affinity
for a broad spectrum of proteins (44). Hence,
addressing the possible target for these ligands is
crucial when assessing the bioactive potential of
food-hydrolysates. Results from the three proteins
with the highest binding affinity with each of the
biopeptides are presented in Table 4.
Table 4. Predicted target proteins for GI-absorbable biopeptides.
Biopeptide
Sequence
Target proteins (Probabilitya )
Protein 1 Protein 2 Protein 3
Val-Leu Calpain-I (0.25) ACE (0.15) COX-2 (0.13)
Ala-Leu COX-2 (0.21) DPP-IV (0.09) HLA-A3 (0.08)
Pro-Leu DPP-IV (0.42) ACE (0.28) Calpain-I (0.19)
Ile-Phe Calpain-I (0.54) ACE (0.50) COX-2 (0.17)
Ala-Phe Calpain-I (0.30) Neprilysin (0.16) SMOT (0.16)
Cys-Phe Calpain-I (0.08) ACE (0.06) Neprilysin (0.06)
Gly-Leu COX-2 (0.13) HLA-A3 (0.10) ACE (0.09)
ACE, Angiotensin Converting Enzyme; COX-2, Cyclooxygenase-2; DPP-IV, Dipeptidyl-Peptidase IV; SMOT, Small Intestine Oligopeptide Transporter; HLA-A3, Human
Leukocyte Antigen-A3. a Probability of the biopeptide to have each protein as a target.
Based on the results obtained from in silico target
prediction analysis, seven suitable proteins were
reported to be likely bonded to GI-absorbable
bioactive peptides. Five of seven biopeptides
exhibited a binding affinity for ACE in line with
their anti-hypertensive properties. Furthermore,
possible interactions with neprilysin were reported
for peptides Ala-Phe and Cys-Phe. Neprilysin is a
Zn2+ -dependent protease tightly involved in the
renin-angiotensin-aldosterone system, particularly
catalyzing natriuretic peptides degradation (45).
Sacubitril (the first neprilysin inhibitor approved)
has shown promising results for hypertension
therapy when administered concomitantly with
angiotensin-receptor blockers (46). Based on the
findings of neprilysin affinity, the results suggest
that biopeptides could modulate vascular resistance
through ACE inhibition and this protease. Further
studies could elucidate these interactions’ feasibility
and therapeutic application.
Interestingly, a high probability of interaction with
cyclooxygenase-2 (COX-2) was also reported.
predicts the probability of achieving a bioavailability
greater than 30% when orally administered (41).
Five of seven peptides scored >50% chance of
surpassing this threshold. Distribution evaluation
revealed that GI-absorbable bioactive peptides are
likely to cross the blood-brain barrier. Based on the
plasma protein binding probability results, peptides
are expected to remain in the bloodstream without
binding to carrying proteins.
Metabolism was assessed based on the interactions
with cytochrome enzymes (CYP450). This large family
of oxidoreductase enzymes are tightly involved in
xenobiotics metabolism; hence, it is important to
predict their interactions with drug-like compounds
(42). Bioactive peptides reported no single substrate-
like or inhibitory interaction with any CYP450. On the
other hand, when assessing toxicity, results show that
peptides induce no liver injury based on the DILI
parameter. Our findings support the hypothesis of
Acquah et al. (8) since in silico evaluation indicated
that biopeptides exert little complications in
humans in terms of toxicity and organ damage. At
last, excretion was evaluated based on half-life and
clearance rate. Half-life varied irregularly among
these peptides, and only three reported a value
higher than 1 hour. On the other hand, clearance
predicted values were all above 1 cm 3 ·min -1
·kg -1
.
These two parameters must be carefully considered,
as low half-life and high clearance could imply
an increased dosing regimen and consumption
frequency to achieve an effective therapeutic effect
(43). This work develops a non-previously analyzed
pharmacokinetic profile of bioactive peptides, yet,
in vivo studies are still imperative to conclude with
a greater degree of certainty.
Target Prediction
Bioactive peptides exhibit a doubtless affinity
for a broad spectrum of proteins (44). Hence,
addressing the possible target for these ligands is
crucial when assessing the bioactive potential of
food-hydrolysates. Results from the three proteins
with the highest binding affinity with each of the
biopeptides are presented in Table 4.
Table 4. Predicted target proteins for GI-absorbable biopeptides.
Biopeptide
Sequence
Target proteins (Probabilitya )
Protein 1 Protein 2 Protein 3
Val-Leu Calpain-I (0.25) ACE (0.15) COX-2 (0.13)
Ala-Leu COX-2 (0.21) DPP-IV (0.09) HLA-A3 (0.08)
Pro-Leu DPP-IV (0.42) ACE (0.28) Calpain-I (0.19)
Ile-Phe Calpain-I (0.54) ACE (0.50) COX-2 (0.17)
Ala-Phe Calpain-I (0.30) Neprilysin (0.16) SMOT (0.16)
Cys-Phe Calpain-I (0.08) ACE (0.06) Neprilysin (0.06)
Gly-Leu COX-2 (0.13) HLA-A3 (0.10) ACE (0.09)
ACE, Angiotensin Converting Enzyme; COX-2, Cyclooxygenase-2; DPP-IV, Dipeptidyl-Peptidase IV; SMOT, Small Intestine Oligopeptide Transporter; HLA-A3, Human
Leukocyte Antigen-A3. a Probability of the biopeptide to have each protein as a target.
Based on the results obtained from in silico target
prediction analysis, seven suitable proteins were
reported to be likely bonded to GI-absorbable
bioactive peptides. Five of seven biopeptides
exhibited a binding affinity for ACE in line with
their anti-hypertensive properties. Furthermore,
possible interactions with neprilysin were reported
for peptides Ala-Phe and Cys-Phe. Neprilysin is a
Zn2+ -dependent protease tightly involved in the
renin-angiotensin-aldosterone system, particularly
catalyzing natriuretic peptides degradation (45).
Sacubitril (the first neprilysin inhibitor approved)
has shown promising results for hypertension
therapy when administered concomitantly with
angiotensin-receptor blockers (46). Based on the
findings of neprilysin affinity, the results suggest
that biopeptides could modulate vascular resistance
through ACE inhibition and this protease. Further
studies could elucidate these interactions’ feasibility
and therapeutic application.
Interestingly, a high probability of interaction with
cyclooxygenase-2 (COX-2) was also reported.
11Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
COX-2 is a rate-limiting enzyme that catalyzes the
production of prostaglandins. Hence, its inhibition
conveys a crucial anti-inflammatory mechanism
shared by non-steroidal anti-inflammatory drugs
(NSAIDs). Several COX-2 inhibitors of natural origin
have been identified, as stated by Cui et al. (47),
yet bioactive peptides have not been reported
among these naturally occurring anti-inflammatory
compounds. Our results and findings of Reyes-
Díaz et al. (48) (who isolated anti-inflammatory
biopeptides from legume proteins) suggest that
bioactive peptides derived from dietary have anti-
inflammatory properties. Therefore, we encourage
a study of those peptides in vitro and in vivo.
Moreover, human leukocyte antigen-A3 (HLA-A3)
is classically associated with hemochromatosis, but
it has also been linked to antiviral T cell-mediated
immune responses (49). Biopeptides Ala-Leu and
Gly-Leu exhibit affinity to HLA-A3, yet the nature
of these interactions remains to be unrevealed. The
small intestine oligopeptide transporter (SMOT)
regulates the peptide absorption from dietary
intake (50) and was reported to be bonded to the
dipeptide Ala-Phe; however, little is known about the
effect of bioactive peptides on the modulation (if not
only a diffusion-channel into enterocytes) of SMOT.
Regarding the anti-diabetic properties of bioactive
peptides, DPP-IV inhibition indicates a high potential
as a glycemic-regulatory agent. Additionally,
Calpain-I was identified as a suitable target protein
for five biopeptides, and four showed the highest
affinity for this enzyme. Calpain-I, a Ca2+ -dependent
cytosolic protease, is pathologically triggered
by alterations in calcium homeostasis commonly
seen in diabetes mellitus (51). These proteases
have been involved in the pathogenesis of several
complications, including diabetic cardiomyopathy
(via activation of the nuclear factor of activated
T-cells (NFAT) and nuclear factor-κB (NF-κB)
which seems to lead to ventricular fibrosis (52)),
diabetic nephropathy (associated with cellular
death and renal dysfunction (53)), and diabetic
retinopathy by induced-proteolysis (causes early
degeneration of cytoskeleton and structure involved
in phototransduction in retinal cells (51)). As a
result, modulators/inhibitors of Calpain-I activity
are proposed as potential therapeutic agents
for type 2 diabetes mellitus (54, 55). Henceforth,
results derived from this investigation indicate that
bioactive peptides could exert a health-promoting
effect in diabetes mellitus aside from potentiating
the incretin effect.
CONCLUSION
Troponin and tropomyosin are rich dietary sources
of bioactive peptides, mainly DPP-IV and ACE
inhibitors. The hydrolysis of these myofibrillar
proteins shows no significant difference (p<0.05)
between the frequency of biopeptides released
from the isoforms of B. taurus, G. gallus, and S.
scrofa. Eight biopeptides were obtained from
simulated gastric and pancreatic digestion of
troponin and tropomyosin: DPP-IV inhibitors, ACE
inhibitors, CaMPDE inhibitors, renin inhibitors, DPP-
III inhibitors, glucose-uptake stimulating, ion-flux
regulating, and antioxidants.
Seven biopeptides from the whole end-products
were classified as absorbable compounds and
exhibited appropriate drug-like properties. The
pharmacokinetic profiling revealed a predicted high
blood-brain barrier diffusion, no CYP450 interaction,
and null toxicity. Even with the pharmacological
properties, the therapeutic potency of biopeptides
is lower than those drugs, indicating that the
nutraceuticals reported in this research must not
replace the current pharmacotherapy. The target
prediction suggests that biopeptides could: [1]
regulate glycemia not only by interacting with DPP-
IV but also with Calpain-I, [2] exert antihypertensive
action by binding to ACE and neprilysin, and [3] hold
anti-inflammatory properties as they reported high
affinity for COX-2. These interactions remain to be
elucidated in future in vitro and in vivo studies.
CONFLICT OF INTEREST
The authors report no conflict of interest.
REFERENCES
1. Ahmad RS, Imran A, Hussain MB. Nutritional Composition of
Meat. In: Arshad MS, editor. Meat Science and Nutrition [Internet].
InTech; 2018 [cited 2021 Jun 16]. Available from: http://www.
intechopen.com/books/meat-science-and-nutrition/nutritional-
composition-of-meat
2. Badui S. Química De Los Alimentos. [Internet]. Pearson Educación
de México, SA de CV; 2006 [cited 2021 Jun 16]. Available
from: ht tps://public.ebookcentral.proquest.com/choice/
publicfullrecord.aspx?p=5133798
3. Premi, M., Bansal, V. Nutraceuticals for Management of Metabolic
Disorders. In: Treating endocrine and metabolic disorders with
herbal medicines. 2021. p. 298–320.
4. Xing L, Liu R, Cao S, Zhang W, Guanghong Z. Meat protein based
bioactive peptides and their potential functional activity: a review.
Int J Food Sci Technol. 2019;54(6):1956–66. DOI: https://doi.
org/10.1111/ijfs.14132
5. Iwaniak A, Minkiewicz P, Pliszka M, Mogut D, Darewicz M.
Characteristics of Biopeptides Released In Silico from Collagens
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
COX-2 is a rate-limiting enzyme that catalyzes the
production of prostaglandins. Hence, its inhibition
conveys a crucial anti-inflammatory mechanism
shared by non-steroidal anti-inflammatory drugs
(NSAIDs). Several COX-2 inhibitors of natural origin
have been identified, as stated by Cui et al. (47),
yet bioactive peptides have not been reported
among these naturally occurring anti-inflammatory
compounds. Our results and findings of Reyes-
Díaz et al. (48) (who isolated anti-inflammatory
biopeptides from legume proteins) suggest that
bioactive peptides derived from dietary have anti-
inflammatory properties. Therefore, we encourage
a study of those peptides in vitro and in vivo.
Moreover, human leukocyte antigen-A3 (HLA-A3)
is classically associated with hemochromatosis, but
it has also been linked to antiviral T cell-mediated
immune responses (49). Biopeptides Ala-Leu and
Gly-Leu exhibit affinity to HLA-A3, yet the nature
of these interactions remains to be unrevealed. The
small intestine oligopeptide transporter (SMOT)
regulates the peptide absorption from dietary
intake (50) and was reported to be bonded to the
dipeptide Ala-Phe; however, little is known about the
effect of bioactive peptides on the modulation (if not
only a diffusion-channel into enterocytes) of SMOT.
Regarding the anti-diabetic properties of bioactive
peptides, DPP-IV inhibition indicates a high potential
as a glycemic-regulatory agent. Additionally,
Calpain-I was identified as a suitable target protein
for five biopeptides, and four showed the highest
affinity for this enzyme. Calpain-I, a Ca2+ -dependent
cytosolic protease, is pathologically triggered
by alterations in calcium homeostasis commonly
seen in diabetes mellitus (51). These proteases
have been involved in the pathogenesis of several
complications, including diabetic cardiomyopathy
(via activation of the nuclear factor of activated
T-cells (NFAT) and nuclear factor-κB (NF-κB)
which seems to lead to ventricular fibrosis (52)),
diabetic nephropathy (associated with cellular
death and renal dysfunction (53)), and diabetic
retinopathy by induced-proteolysis (causes early
degeneration of cytoskeleton and structure involved
in phototransduction in retinal cells (51)). As a
result, modulators/inhibitors of Calpain-I activity
are proposed as potential therapeutic agents
for type 2 diabetes mellitus (54, 55). Henceforth,
results derived from this investigation indicate that
bioactive peptides could exert a health-promoting
effect in diabetes mellitus aside from potentiating
the incretin effect.
CONCLUSION
Troponin and tropomyosin are rich dietary sources
of bioactive peptides, mainly DPP-IV and ACE
inhibitors. The hydrolysis of these myofibrillar
proteins shows no significant difference (p<0.05)
between the frequency of biopeptides released
from the isoforms of B. taurus, G. gallus, and S.
scrofa. Eight biopeptides were obtained from
simulated gastric and pancreatic digestion of
troponin and tropomyosin: DPP-IV inhibitors, ACE
inhibitors, CaMPDE inhibitors, renin inhibitors, DPP-
III inhibitors, glucose-uptake stimulating, ion-flux
regulating, and antioxidants.
Seven biopeptides from the whole end-products
were classified as absorbable compounds and
exhibited appropriate drug-like properties. The
pharmacokinetic profiling revealed a predicted high
blood-brain barrier diffusion, no CYP450 interaction,
and null toxicity. Even with the pharmacological
properties, the therapeutic potency of biopeptides
is lower than those drugs, indicating that the
nutraceuticals reported in this research must not
replace the current pharmacotherapy. The target
prediction suggests that biopeptides could: [1]
regulate glycemia not only by interacting with DPP-
IV but also with Calpain-I, [2] exert antihypertensive
action by binding to ACE and neprilysin, and [3] hold
anti-inflammatory properties as they reported high
affinity for COX-2. These interactions remain to be
elucidated in future in vitro and in vivo studies.
CONFLICT OF INTEREST
The authors report no conflict of interest.
REFERENCES
1. Ahmad RS, Imran A, Hussain MB. Nutritional Composition of
Meat. In: Arshad MS, editor. Meat Science and Nutrition [Internet].
InTech; 2018 [cited 2021 Jun 16]. Available from: http://www.
intechopen.com/books/meat-science-and-nutrition/nutritional-
composition-of-meat
2. Badui S. Química De Los Alimentos. [Internet]. Pearson Educación
de México, SA de CV; 2006 [cited 2021 Jun 16]. Available
from: ht tps://public.ebookcentral.proquest.com/choice/
publicfullrecord.aspx?p=5133798
3. Premi, M., Bansal, V. Nutraceuticals for Management of Metabolic
Disorders. In: Treating endocrine and metabolic disorders with
herbal medicines. 2021. p. 298–320.
4. Xing L, Liu R, Cao S, Zhang W, Guanghong Z. Meat protein based
bioactive peptides and their potential functional activity: a review.
Int J Food Sci Technol. 2019;54(6):1956–66. DOI: https://doi.
org/10.1111/ijfs.14132
5. Iwaniak A, Minkiewicz P, Pliszka M, Mogut D, Darewicz M.
Characteristics of Biopeptides Released In Silico from Collagens
12Journal Vitae | https://revistas.udea.edu.co/index.php/vitae Volume 30 | Number 01 | Article 347310Jorge Andrés Barrero, María Alejandra Barrero-Casallas, Angélica María González-Clavijo, Marcela Cruz-González
Using Quantitative Parameters. Foods. 2020;9(7):965. DOI:
https://doi.org/10.3390/foods9070965
6. Wang TY, Hsieh CH, Hung CC, Jao CL, Lin PY, Hsieh YL, et al.
A study to evaluate the potential of an in silico approach for
predicting dipeptidyl peptidase-IV inhibitory activity in vitro of
protein hydrolysates. Food Chem. 2017;234:431–8. DOI: https://
doi.org/10.1016/j.foodchem.2017.05.035
7. Jao C-L, Hung C-C, Tung Y-S, Lin P-Y, Chen M-C, Hsu K-C.
The development of bioactive peptides from dietary proteins
as a dipeptidyl peptidase IV inhibitor for the management of
type 2 diabetes. BioMedicine. 2015;5(3):14. DOI: https://doi.
org/10.7603/s40681-015-0014-9
8. Acquah C, Dzuvor CKO, Tosh S, Agyei D. Anti-diabetic effects
of bioactive peptides: recent advances and clinical implications.
Crit Rev Food Sci Nutr. 2020;1–14. DOI: https://doi.org/10.1080
/10408398.2020.1851168
9. Tu M, Cheng S, Lu W, Du M. Advancement and prospects of
bioinformatics analysis for studying bioactive peptides from
food-derived protein: Sequence, structure, and functions. TrAC
Trends Anal Chem. 2018;105:7–17. DOI: https://doi.org/10.1016/j.
trac.2018.04.005
10. Dong J, Wang N-N, Yao Z-J, Zhang L, Cheng Y, Ouyang D, et
al. ADMETlab: a platform for systematic ADMET evaluation
based on a comprehensively collected ADMET database. J
Cheminformatics. 2018;10(1):29. DOI: https://doi.org/10.1186/
s13321-018-0283-x
11. Daina A, Michielin O, Zoete V. SwissTargetPrediction: updated
data and new features for efficient prediction of protein targets
of small molecules. Nucleic Acids Res. 2019;47(W1):W357–64.
DOI: https://doi.org/10.1093/nar/gkz382
12. The UniProt Consortium. UniProt: a worldwide hub of protein
knowledge. Nucleic Acids Res. 2019;47(D1):D506–15. DOI: https://
doi.org/10.1093/nar/gky1049
13. Minkiewicz, Iwaniak, Darewicz. BIOPEP-UWM Database of
Bioactive Peptides: Current Opportunities. Int J Mol Sci.
2019;20(23):5978. DOI: https://doi.org/10.3390/ijms20235978
14. Daina A, Michielin O, Zoete V. SwissADME: a free web tool
to evaluate pharmacokinetics, drug-likeness and medicinal
chemistry friendliness of small molecules. Sci Rep. 2017;7(1):42717.
DOI: https://doi.org/10.1038/srep42717
15. Lee Y-S, Jun H-S. Anti-diabetic actions of glucagon-like peptide-1
on pancreatic beta-cells. Metabolism. 2014;63(1):9–19. DOI:
https://doi.org/10.1016/j.metabol.2013.09.010
16. Casanova-Martí À, Bravo FI, Serrano J, Ardévol A, Pinent
M, Muguerza B. Antihyperglycemic effect of a chicken feet
hydrolysate via the incretin system: DPP-IV-inhibitory activity and
GLP-1 release stimulation. Food Funct. 2019;10(7):4062–70. DOI:
https://doi.org/10.1039/C9FO00695H
17. Soler MJ, Batlle D. Revisiting the renin-angiotensin system. Mol
Cell Endocrinol. 2021;529:111268. DOI: https://doi.org/10.1016/j.
mce.2021.111268
18. Martin M, Deussen A. Effects of natural peptides from food
proteins on angiotensin conver ting enzyme activity and
hypertension. Crit Rev Food Sci Nutr. 2019;59(8):1264–83. DOI:
https://doi.org/10.1080/10408398.2017.1402750
19. Suetsuna K, Ukeda H, Ochi H. Isolation and characterization of
free radical scavenging activities peptides derived from casein.
J Nutr Biochem. 2000;11(3):128–31. DOI: https://doi.org/10.1016/
S0955-2863(99)00083-2
20. Liu C, Ren D, Li J, Fang L, Wang J, Liu J, et al. Cytoprotective effect
and purification of novel antioxidant peptides from hazelnut
(C. heterophylla Fisch) protein hydrolysates. J Funct Foods.
2018;42:203–15. DOI: https://doi.org/10.1016/j.jff.2017.12.003
21. Deniau B, Rehfeld L, Santos K, Dienelt A, Azibani F, Sadoune M, et
al. Circulating dipeptidyl peptidase 3 is a myocardial depressant
factor: dipeptidyl peptidase 3 inhibition rapidly and sustainably
improves haemodynamics. Eur J Heart Fail. 2020;22(2):290–9.
DOI: https://doi.org/10.1002/ejhf.1601
22. Morifuji M, Koga J, Kawanaka K, Higuchi M. Branched-Chain
Amino Acid-Containing Dipeptides, Identified from Whey Protein
Hydrolysates, Stimulate Glucose Uptake Rate in L6 Myotubes
and Isolated Skeletal Muscles. J Nutr Sci Vitaminol (Tokyo).
2009;55(1):81–6. DOI: https://doi.org/10.3177/jnsv.55.81
23. Goraya TA, Cooper DMF. Ca2+- calmodulin- dependent
phosphodies ter ase (PDE1): Cur rent per spec tives. Cell
Signal. 2005;17(7):789–97. DOI: https://doi.org/10.1016/j.
cellsig.2004.12.017
24. O’Brien JJ, O’Callaghan JP, Miller DB, Chalgeri S, Wennogle
LP, Davis RE, et al. Inhibition of calcium-calmodulin-dependent
phosphodiesterase (PDE1) suppresses inflammatory responses.
Mol Cell Neurosci. 2020;102:103449. DOI: ht tps://doi.
org/10.1016/j.mcn.2019.103449
25. Ramya K, Suresh R, Kumar HY, Kumar BRP, Mur thy NBS.
Decades-old renin inhibitors are still struggling to find a niche
in antihypertensive therapy. A fleeting look at the old and the
promising new molecules. Bioorg Med Chem. 2020;28(10):115466.
DOI: https://doi.org/10.1016/j.bmc.2020.115466
26. Ignat’ev DA, Vorob’ev VV, Ziganshin RKh. Effects of a number of
short peptides isolated from the brain of the hibernating ground
squirrel on the eeg and behavior in rats. Neurosci Behav Physiol.
1998;28(2):158–66. DOI: https://doi.org/10.1007/BF02461962
27. Martini S, Conte A, Tagliazucchi D. Comparative peptidomic
profile and bioac tivities of cooked beef, pork, chicken
and turkey meat after in vitro gastro-intestinal digestion. J
Proteomics. 2019;208:103500. DOI: https://doi.org/10.1016/j.
jprot.2019.103500
28. Mora L, Gallego M, Toldrá F. ACEI-Inhibitor y Peptides
Naturally Generated in Meat and Meat Products and Their
Health Relevance. Nutrients. 2018;10(9):1259. DOI: https://doi.
org/10.3390/nu10091259
29. Lan VTT, Ito K, Ohno M, Motoyama T, Ito S, Kawarasaki Y.
Analyzing a dipeptide library to identify human dipeptidyl
peptidase IV inhibitor. Food Chem. 2015;175:66–73. DOI: https://
doi.org/10.1016/j.foodchem.2014.11.131
30. Nongonierma AB, Mooney C, Shields DC, FitzGerald RJ.
Inhibition of dipeptidyl peptidase IV and xanthine oxidase by
amino acids and dipeptides. Food Chem. 2013;141(1):644–53.
DOI: https://doi.org/10.1016/j.foodchem.2013.02.115
31. Byun H-G, Kim S-K. Structure and Activity of Angiotensin I
Converting Enzyme Inhibitory Peptides Derived from Alaskan
Pollack Skin. BMB Rep. 2002;35(2):239–43. DOI: https://doi.
org/10.5483/BMBRep.2002.35.2.239
32. Cheung, H. S., Wang, F. L., Ondetti, M. A., Sabo, E. F., Cushman,
D. W. Binding of peptide substrates and inhibitors of angiotensin-
converting enzyme. Importance of the COOH-terminal dipeptide
sequence. J Biol Chem [Internet]. 1980;255(2). Available from:
https://doi.org/pubmed.ncbi.nlm.nih.gov/6243277/
33. Wu H, He H-L, Chen X-L, Sun C-Y, Zhang Y-Z, Zhou B-C.
Purification and identification of novel angiotensin-I-converting
enzyme inhibitory peptides from shark meat hydrolysate. Process
Biochem. 2008;43(4):457–61. DOI: https://doi.org/10.1016/j.
procbio.2008.01.018
34. Ryan JT, Ross RP, Bolton D, Fitzgerald GF, Stanton C. Bioactive
Peptides from Muscle Sources: Meat and Fish. Nutrients.
2011;3(9):765–91. DOI: https://doi.org/10.3390/nu3090765
35. Keller F, Hartmann B, Czock D. Time of effect duration and
administration interval for sitagliptin in patients with kidney
Using Quantitative Parameters. Foods. 2020;9(7):965. DOI:
https://doi.org/10.3390/foods9070965
6. Wang TY, Hsieh CH, Hung CC, Jao CL, Lin PY, Hsieh YL, et al.
A study to evaluate the potential of an in silico approach for
predicting dipeptidyl peptidase-IV inhibitory activity in vitro of
protein hydrolysates. Food Chem. 2017;234:431–8. DOI: https://
doi.org/10.1016/j.foodchem.2017.05.035
7. Jao C-L, Hung C-C, Tung Y-S, Lin P-Y, Chen M-C, Hsu K-C.
The development of bioactive peptides from dietary proteins
as a dipeptidyl peptidase IV inhibitor for the management of
type 2 diabetes. BioMedicine. 2015;5(3):14. DOI: https://doi.
org/10.7603/s40681-015-0014-9
8. Acquah C, Dzuvor CKO, Tosh S, Agyei D. Anti-diabetic effects
of bioactive peptides: recent advances and clinical implications.
Crit Rev Food Sci Nutr. 2020;1–14. DOI: https://doi.org/10.1080
/10408398.2020.1851168
9. Tu M, Cheng S, Lu W, Du M. Advancement and prospects of
bioinformatics analysis for studying bioactive peptides from
food-derived protein: Sequence, structure, and functions. TrAC
Trends Anal Chem. 2018;105:7–17. DOI: https://doi.org/10.1016/j.
trac.2018.04.005
10. Dong J, Wang N-N, Yao Z-J, Zhang L, Cheng Y, Ouyang D, et
al. ADMETlab: a platform for systematic ADMET evaluation
based on a comprehensively collected ADMET database. J
Cheminformatics. 2018;10(1):29. DOI: https://doi.org/10.1186/
s13321-018-0283-x
11. Daina A, Michielin O, Zoete V. SwissTargetPrediction: updated
data and new features for efficient prediction of protein targets
of small molecules. Nucleic Acids Res. 2019;47(W1):W357–64.
DOI: https://doi.org/10.1093/nar/gkz382
12. The UniProt Consortium. UniProt: a worldwide hub of protein
knowledge. Nucleic Acids Res. 2019;47(D1):D506–15. DOI: https://
doi.org/10.1093/nar/gky1049
13. Minkiewicz, Iwaniak, Darewicz. BIOPEP-UWM Database of
Bioactive Peptides: Current Opportunities. Int J Mol Sci.
2019;20(23):5978. DOI: https://doi.org/10.3390/ijms20235978
14. Daina A, Michielin O, Zoete V. SwissADME: a free web tool
to evaluate pharmacokinetics, drug-likeness and medicinal
chemistry friendliness of small molecules. Sci Rep. 2017;7(1):42717.
DOI: https://doi.org/10.1038/srep42717
15. Lee Y-S, Jun H-S. Anti-diabetic actions of glucagon-like peptide-1
on pancreatic beta-cells. Metabolism. 2014;63(1):9–19. DOI:
https://doi.org/10.1016/j.metabol.2013.09.010
16. Casanova-Martí À, Bravo FI, Serrano J, Ardévol A, Pinent
M, Muguerza B. Antihyperglycemic effect of a chicken feet
hydrolysate via the incretin system: DPP-IV-inhibitory activity and
GLP-1 release stimulation. Food Funct. 2019;10(7):4062–70. DOI:
https://doi.org/10.1039/C9FO00695H
17. Soler MJ, Batlle D. Revisiting the renin-angiotensin system. Mol
Cell Endocrinol. 2021;529:111268. DOI: https://doi.org/10.1016/j.
mce.2021.111268
18. Martin M, Deussen A. Effects of natural peptides from food
proteins on angiotensin conver ting enzyme activity and
hypertension. Crit Rev Food Sci Nutr. 2019;59(8):1264–83. DOI:
https://doi.org/10.1080/10408398.2017.1402750
19. Suetsuna K, Ukeda H, Ochi H. Isolation and characterization of
free radical scavenging activities peptides derived from casein.
J Nutr Biochem. 2000;11(3):128–31. DOI: https://doi.org/10.1016/
S0955-2863(99)00083-2
20. Liu C, Ren D, Li J, Fang L, Wang J, Liu J, et al. Cytoprotective effect
and purification of novel antioxidant peptides from hazelnut
(C. heterophylla Fisch) protein hydrolysates. J Funct Foods.
2018;42:203–15. DOI: https://doi.org/10.1016/j.jff.2017.12.003
21. Deniau B, Rehfeld L, Santos K, Dienelt A, Azibani F, Sadoune M, et
al. Circulating dipeptidyl peptidase 3 is a myocardial depressant
factor: dipeptidyl peptidase 3 inhibition rapidly and sustainably
improves haemodynamics. Eur J Heart Fail. 2020;22(2):290–9.
DOI: https://doi.org/10.1002/ejhf.1601
22. Morifuji M, Koga J, Kawanaka K, Higuchi M. Branched-Chain
Amino Acid-Containing Dipeptides, Identified from Whey Protein
Hydrolysates, Stimulate Glucose Uptake Rate in L6 Myotubes
and Isolated Skeletal Muscles. J Nutr Sci Vitaminol (Tokyo).
2009;55(1):81–6. DOI: https://doi.org/10.3177/jnsv.55.81
23. Goraya TA, Cooper DMF. Ca2+- calmodulin- dependent
phosphodies ter ase (PDE1): Cur rent per spec tives. Cell
Signal. 2005;17(7):789–97. DOI: https://doi.org/10.1016/j.
cellsig.2004.12.017
24. O’Brien JJ, O’Callaghan JP, Miller DB, Chalgeri S, Wennogle
LP, Davis RE, et al. Inhibition of calcium-calmodulin-dependent
phosphodiesterase (PDE1) suppresses inflammatory responses.
Mol Cell Neurosci. 2020;102:103449. DOI: ht tps://doi.
org/10.1016/j.mcn.2019.103449
25. Ramya K, Suresh R, Kumar HY, Kumar BRP, Mur thy NBS.
Decades-old renin inhibitors are still struggling to find a niche
in antihypertensive therapy. A fleeting look at the old and the
promising new molecules. Bioorg Med Chem. 2020;28(10):115466.
DOI: https://doi.org/10.1016/j.bmc.2020.115466
26. Ignat’ev DA, Vorob’ev VV, Ziganshin RKh. Effects of a number of
short peptides isolated from the brain of the hibernating ground
squirrel on the eeg and behavior in rats. Neurosci Behav Physiol.
1998;28(2):158–66. DOI: https://doi.org/10.1007/BF02461962
27. Martini S, Conte A, Tagliazucchi D. Comparative peptidomic
profile and bioac tivities of cooked beef, pork, chicken
and turkey meat after in vitro gastro-intestinal digestion. J
Proteomics. 2019;208:103500. DOI: https://doi.org/10.1016/j.
jprot.2019.103500
28. Mora L, Gallego M, Toldrá F. ACEI-Inhibitor y Peptides
Naturally Generated in Meat and Meat Products and Their
Health Relevance. Nutrients. 2018;10(9):1259. DOI: https://doi.
org/10.3390/nu10091259
29. Lan VTT, Ito K, Ohno M, Motoyama T, Ito S, Kawarasaki Y.
Analyzing a dipeptide library to identify human dipeptidyl
peptidase IV inhibitor. Food Chem. 2015;175:66–73. DOI: https://
doi.org/10.1016/j.foodchem.2014.11.131
30. Nongonierma AB, Mooney C, Shields DC, FitzGerald RJ.
Inhibition of dipeptidyl peptidase IV and xanthine oxidase by
amino acids and dipeptides. Food Chem. 2013;141(1):644–53.
DOI: https://doi.org/10.1016/j.foodchem.2013.02.115
31. Byun H-G, Kim S-K. Structure and Activity of Angiotensin I
Converting Enzyme Inhibitory Peptides Derived from Alaskan
Pollack Skin. BMB Rep. 2002;35(2):239–43. DOI: https://doi.
org/10.5483/BMBRep.2002.35.2.239
32. Cheung, H. S., Wang, F. L., Ondetti, M. A., Sabo, E. F., Cushman,
D. W. Binding of peptide substrates and inhibitors of angiotensin-
converting enzyme. Importance of the COOH-terminal dipeptide
sequence. J Biol Chem [Internet]. 1980;255(2). Available from:
https://doi.org/pubmed.ncbi.nlm.nih.gov/6243277/
33. Wu H, He H-L, Chen X-L, Sun C-Y, Zhang Y-Z, Zhou B-C.
Purification and identification of novel angiotensin-I-converting
enzyme inhibitory peptides from shark meat hydrolysate. Process
Biochem. 2008;43(4):457–61. DOI: https://doi.org/10.1016/j.
procbio.2008.01.018
34. Ryan JT, Ross RP, Bolton D, Fitzgerald GF, Stanton C. Bioactive
Peptides from Muscle Sources: Meat and Fish. Nutrients.
2011;3(9):765–91. DOI: https://doi.org/10.3390/nu3090765
35. Keller F, Hartmann B, Czock D. Time of effect duration and
administration interval for sitagliptin in patients with kidney
13Journal Vitae | https://revistas.udea.edu.co/index.php/vitaeVolume 30 | Number 01 | Article 347310
Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
failure. Eur J Drug Metab Pharmacokinet. 2014;39(2):77–85. DOI:
https://doi.org/10.1007/s13318-013-0164-7
36. Lin Y, Feng M, Lu C-W, Lei Y-P, He Z-M, Xiong Y. Preservation of
vascular DDAH activity contributes to the protection of captopril
against endothelial dysfunction in hyperlipidemic rabbits. Eur
J Pharmacol. 2017;798:43–8. DOI: https://doi.org/10.1016/j.
ejphar.2017.01.041
37. Doroschuk VO, Makukha OG. The peculiarities of the interphase
distribution of amino acids in the cloud point extraction systems.
Colloids Surf Physicochem Eng Asp. 2017;520:757–63. DOI:
https://doi.org/10.1016/j.colsurfa.2017.02.036
38. Das T, Meht a CH, Nayak U Y. Multiple approaches for
achieving drug solubility: an in silico perspective. Drug Discov
Today. 2020;25(7):1206 –12. DOI: https://doi.org/10.1016/j.
drudis.2020.04.016
39. Acquah C, Stefano ED, Udenigwe CC. Role of hydrophobicity
in food peptide functionality and bioactivity. J Food Bioact.
2018;4(1):88–98. DOI: https://doi.org/10.31665/JFB.2018.4164
40. Yosipof A, Guedes RC, García-Sosa AT. Data Mining and Machine
Learning Models for Predicting Drug Likeness and Their Disease
or Organ Category. Front Chem. 2018;6:162. DOI: https://doi.
org/10.3389/fchem.2018.00162
41. Tian S, Li Y, Wang J, Zhang J, Hou T. ADME Evaluation in Drug
Discovery. 9. Prediction of Oral Bioavailability in Humans Based
on Molecular Properties and Structural Fingerprints. Mol Pharm.
2011;8(3):841–51. DOI: https://doi.org/10.1021/mp100444g
42. Isvoran A, Louet M, Vladoiu DL, Craciun D, Loriot M-A, Villoutreix
BO, et al. Pharmacogenomics of the cytochrome P450 2C family:
impacts of amino acid variations on drug metabolism. Drug
Discov Today. 2017;22(2):366–76. DOI: https://doi.org/10.1016/j.
drudis.2016.09.015
43. Smith DA, Beaumont K, Maurer TS, Di L. Relevance of Half-Life
in Drug Design. J Med Chem. 2018;61(10):4273–82. DOI: https://
doi.org/10.1021/acs.jmedchem.7b00969
44. Iwaniak A, Minkiewicz P, Darewicz M, Hrynkiewicz M. Food
protein-originating peptides as tastants - Physiological,
technological, sensor y, and bioinfor matic approaches.
Food Res Int. 2016;89:27–38. DOI: https://doi.org/10.1016/j.
foodres.2016.08.010
45. Bayes-Genis A, Barallat J, Richards AM. A Test in Context:
Neprilysin. J Am Coll Cardiol. 2016;68(6):639–53. DOI: https://
doi.org/10.1016/j.jacc.2016.04.060
46. Salazar J, Rojas-Quintero J, Cano C, Pérez JL, Ramírez P,
Carrasquero R, et al. Neprilysin: A Potential Therapeutic Target
of Arterial Hypertension? Curr Cardiol Rev. 2020;16(1):25–35.
DOI: https://doi.org/10.2174/1573403X15666190625160352
47. Cui J, Jia J. Natural COX-2 Inhibitors as Promising Anti-inflammatory
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https://doi.org/10.2174/0929867327999200917150939
48. Reyes-Díaz A, Del-Toro-Sánchez CL, Rodríguez-Figueroa JC,
Valdéz-Hurtado S, Wong-Corral FJ, Borboa-Flores J, et al.
Legume Proteins as a Promising Source of Anti-Inflammatory
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Fast Skeletal Muscle Troponin and Tropomyosin as a Dietary Source of Antidiabetic and Antihypertensive Bioactive Peptides: An In Silico Study
failure. Eur J Drug Metab Pharmacokinet. 2014;39(2):77–85. DOI:
https://doi.org/10.1007/s13318-013-0164-7
36. Lin Y, Feng M, Lu C-W, Lei Y-P, He Z-M, Xiong Y. Preservation of
vascular DDAH activity contributes to the protection of captopril
against endothelial dysfunction in hyperlipidemic rabbits. Eur
J Pharmacol. 2017;798:43–8. DOI: https://doi.org/10.1016/j.
ejphar.2017.01.041
37. Doroschuk VO, Makukha OG. The peculiarities of the interphase
distribution of amino acids in the cloud point extraction systems.
Colloids Surf Physicochem Eng Asp. 2017;520:757–63. DOI:
https://doi.org/10.1016/j.colsurfa.2017.02.036
38. Das T, Meht a CH, Nayak U Y. Multiple approaches for
achieving drug solubility: an in silico perspective. Drug Discov
Today. 2020;25(7):1206 –12. DOI: https://doi.org/10.1016/j.
drudis.2020.04.016
39. Acquah C, Stefano ED, Udenigwe CC. Role of hydrophobicity
in food peptide functionality and bioactivity. J Food Bioact.
2018;4(1):88–98. DOI: https://doi.org/10.31665/JFB.2018.4164
40. Yosipof A, Guedes RC, García-Sosa AT. Data Mining and Machine
Learning Models for Predicting Drug Likeness and Their Disease
or Organ Category. Front Chem. 2018;6:162. DOI: https://doi.
org/10.3389/fchem.2018.00162
41. Tian S, Li Y, Wang J, Zhang J, Hou T. ADME Evaluation in Drug
Discovery. 9. Prediction of Oral Bioavailability in Humans Based
on Molecular Properties and Structural Fingerprints. Mol Pharm.
2011;8(3):841–51. DOI: https://doi.org/10.1021/mp100444g
42. Isvoran A, Louet M, Vladoiu DL, Craciun D, Loriot M-A, Villoutreix
BO, et al. Pharmacogenomics of the cytochrome P450 2C family:
impacts of amino acid variations on drug metabolism. Drug
Discov Today. 2017;22(2):366–76. DOI: https://doi.org/10.1016/j.
drudis.2016.09.015
43. Smith DA, Beaumont K, Maurer TS, Di L. Relevance of Half-Life
in Drug Design. J Med Chem. 2018;61(10):4273–82. DOI: https://
doi.org/10.1021/acs.jmedchem.7b00969
44. Iwaniak A, Minkiewicz P, Darewicz M, Hrynkiewicz M. Food
protein-originating peptides as tastants - Physiological,
technological, sensor y, and bioinfor matic approaches.
Food Res Int. 2016;89:27–38. DOI: https://doi.org/10.1016/j.
foodres.2016.08.010
45. Bayes-Genis A, Barallat J, Richards AM. A Test in Context:
Neprilysin. J Am Coll Cardiol. 2016;68(6):639–53. DOI: https://
doi.org/10.1016/j.jacc.2016.04.060
46. Salazar J, Rojas-Quintero J, Cano C, Pérez JL, Ramírez P,
Carrasquero R, et al. Neprilysin: A Potential Therapeutic Target
of Arterial Hypertension? Curr Cardiol Rev. 2020;16(1):25–35.
DOI: https://doi.org/10.2174/1573403X15666190625160352
47. Cui J, Jia J. Natural COX-2 Inhibitors as Promising Anti-inflammatory
Agents: An Update. Curr Med Chem. 2021;28(18):3622–46. DOI:
https://doi.org/10.2174/0929867327999200917150939
48. Reyes-Díaz A, Del-Toro-Sánchez CL, Rodríguez-Figueroa JC,
Valdéz-Hurtado S, Wong-Corral FJ, Borboa-Flores J, et al.
Legume Proteins as a Promising Source of Anti-Inflammatory
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