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Cubeb (Piper cubeba L.f.): A comprehensive review of its botany, phytochemistry, traditional uses, and pharmacological properties

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Piper cubeba L.f. (Piperaceae), known as cubeb, is a popular traditional herbal medicine used for the treatment of many diseases, especially digestive and respiratory disorders. The plant is rich in essential oil, found mainly in fruits, and this makes it economically important. Many traditional utilizations have been also validated from the plant and its isolated compounds owing to their antioxidant, antibacterial, anti-inflammatory and anticancer effects. These biological activities are attributed to the phytochemicals (phenolic compounds, lignans and alkaloids) and the essential oil of the plant. The present work aims to provide an up-to-date review on the traditional uses, phytochemistry and pharmacology of the plant and discusses the future perspectives to promote its valorization for nutritional- and health-promoting effects.
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fnut-09-1048520 November 16, 2022 Time: 14:20 # 1
TYPE Review
PUBLISHED 22 November 2022
DOI 10.3389/fnut.2022.1048520
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EDITED BY
Rajeev K. Singla,
Sichuan University, China
REVIEWED BY
Lee Seong Wei,
Universiti Malaysia Kelantan, Malaysia
Anindita Chakravarti,
Maharani Kasiswari College, India
Vineet Mittal,
Maharshi Dayanand University, India
*CORRESPONDENCE
Mansour Sobeh
mansour.sobeh@um6p.ma
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Frontiers in Nutrition
RECEIVED 19 September 2022
ACCEPTED 01 November 2022
PUBLISHED 22 November 2022
CITATION
Drissi B, Mahdi I, Yassir M,
Ben Bakrim W, Bouissane L and
Sobeh M (2022) Cubeb (Piper cubeba
L.f.): A comprehensive review of its
botany, phytochemistry,
traditional uses, and pharmacological
properties.
Front. Nutr. 9:1048520.
doi: 10.3389/fnut.2022.1048520
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© 2022 Drissi, Mahdi, Yassir, Ben
Bakrim, Bouissane and Sobeh. This is
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does not comply with these terms.
Cubeb (Piper cubeba L.f.): A
comprehensive review of its
botany, phytochemistry,
traditional uses, and
pharmacological properties
Badreddine Drissi1, Ismail Mahdi1, Mouna Yassir1,
Widad Ben Bakrim1,2 , Latifa Bouissane3and Mansour Sobeh1*
1AgroBioSciences, Mohammed VI Polytechnic University, Ben-Guerir, Morocco, 2African
Sustainable Agriculture Research Institute (ASARI), Mohammed VI Polytechnic University, Laayoune,
Morocco, 3Molecular Chemistry, Materials and Catalysis Laboratory, Faculty of Sciences and
Technologies, Sultan Moulay Slimane University, Beni-Mellal, Morocco
Piper cubeba L.f. (Piperaceae), known as cubeb, is a popular traditional
herbal medicine used for the treatment of many diseases, especially digestive
and respiratory disorders. The plant is rich in essential oil, found mainly in
fruits, and this makes it economically important. Many traditional utilizations
have been also validated from the plant and its isolated compounds owing
to their antioxidant, antibacterial, anti-inflammatory and anticancer effects.
These biological activities are attributed to the phytochemicals (phenolic
compounds, lignans and alkaloids) and the essential oil of the plant. The
present work aims to provide an up-to-date review on the traditional
uses, phytochemistry and pharmacology of the plant and discusses the
future perspectives to promote its valorization for nutritional- and health-
promoting effects.
KEYWORDS
cubeb, Piper cubeba, phytochemistry, traditional uses, pharmacological activities
Introduction
Aromatic and medicinal plants (AMPs) have been used since antiquity as therapeutic
and cosmeceutical agents (1). In addition, the vast ethnopharmacological applications
of AMPs have inspired the current research to provide and discover new main drugs
against various health disorders (2). The genus Piper belongs to the family Piperaceae
which has more than 700 species. They are both erect and spreading herbs, shrubs,
or rarely trees with great economic and medicinal importance (3). Piper cubeba is a
native plant of Java and Borneo where the appellation of this plant is the Java pepper.
It is one of the plants of the folk pepiraceae species that is used as a spice. The
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plant is cultivated for its berries, which are rich in essential oil
(4). Economically, the plant is an important source of its dried
berries as they have several applications in perfumes, cosmetics,
and food preservatives (5). In Moroccan cuisine, cubeb is
popular in savory dishes and pastries such as markouts and the
famous Ras el hanout spice blend (a popular mixture of herbs
and spices used throughout the Middle East and North Africa)
(6). Cubeb is marketed by a Swiss company as a refreshing agent
and used in various products such as chewing gums, alcoholic
and soft drinks, sherbets, gelatin confectionery, and toothpaste
(6). Cubeb is also used to flavor alcoholic and non-alcoholic
drinks such as Bombay Sapphire gin and Pertsovka, a Russian
pepper vodka which is prepared from a cubeb infusion (6).
Traditionally, P. cubeba is used for the treatment of gonorrhea,
dysentery, syphilis, abdominal pain, diarrhea, enteritis, and
asthmatic diseases (5). The plant possesses also outstanding
pharmacological activities. For instance, P. cubebas essential oil
furnished antiparasitic, antimicrobial, and insecticidal activities
(7). In addition, different extracts from the plant demonstrated
antioxidant, antimicrobial, nephroprotetive, hepatoprotective,
and anti-inflammatory activities (8). These biological activities
are due to its chemical composition, especially, phenolic acids
and flavonoids, that have been detected in P. cubeba extracts
(911). The plant is also a rich source of lignans particularly
cubebin, a bioactive compound with a wide range of biological
activities such as antimicrobial, anticaner, and neuroprotective,
among others (5,12). Overall, the most reported modes
of action by which P. cubeba extracts exert its biological
activities involve many intracellular targets, among them the
regulation of genes expression, inhibition of oxidative stress,
induction of apoptosis and quorum sensing inhibition in
pathogenic microbes.
The present review aims to collect and collate the available
literature on the botany of plant and provide an insight
about its chemical composition and diverse biological activities
including antioxidant, anti-inflammatory, antibacterial, wound
healing, antidiabetic, and renoprotective activities as well as its
agricultural applications. It also highlights future perspectives to
further maximize the exploitation of the plant in nutraceuticals,
cosmeceuticals, and food applications.
Literature research
The literature search was conducted through the Web
of science, Scopus, PubMed, SciFinder, and other databases.
The keywords used included Piper cubeba, “chemical
composition, “pharmacological properties, and “biological
activities.” Information has been collected from relevant
textbook, reviews, and documents. Duplicated and irrelevant
works were excluded as well as non-English documents, and
those with unavailable full text (Figure 1). The species name
was checked based on the online database.1Various types of
information regarding P. cubeba are discussed in corresponding
parts of the paper.
Morphological and geographical
description
P. cubeba is one of the most popular species of the
Piperaceae family and the most widespread population of this
species is generally found in Indonesia, India, medieval Europe,
and North Africa (Figure 2) (5,13). Cubeb is a woody climbing
perennial that has stem and ashy-grey climbing branches. The
length is 5–15 m high. The leaves are ovate with cordate or
rounded base, glabrous with a thick pedicle, simple, smooth,
and pointed at the apex, the lower surface is densely provided
with tiny glands embedded. They are completely margined,
tough and up to 15 cm long and 6 cm wide (Figure 3A) (5,
9). The flowers are small, dense unisexual that are glued to the
peduncles, arranged in 4 cm long scaly spikes that have 2–3
stamens. The female tips have about 50 individual flowers with
an ovary of 4 carpels fused with 4 sessile stigmas. Flowering takes
place in winter (Figure 3B) (5). The fruits are globose from 6 to
8 mm in diameter. The upper part of the fruit has a diameter of
3–6 mm and covered by grayish brown, pericarp that extends
at the base into a straight stem. They have a spicy, aromatic
smell and a bitter taste. The fruit has a single dark brown
sub-globose seed with a width of 3–4 mm (9) (Figure 3C).
The plant has different vernacular names depending on its
distribution (Table 1).
Phytochemical composition
Piper species are characterized by the production of typical
phytochemical compounds such as benzoic acids, amides,
chromenos, terpenes, phenylpropanoids, lignans, alkaloids, fatty
acids, and hydrocarbons (14). The alkaloid piperine and the
two lignans cubebin and hinokinin are the most abundant
compounds from the berries (15).
Lignans
Altogether, 28 lignans were annotated from P. cubeba
(leaves, berries, stalks) using GC, GC-MS, HPLC, and NMR.
Out of which, 4 lignans were detected in all plant parts
(leaves, berries, stalks), 9 lignans were found only in the leaves
and berries, 2 lignans were solely characterized from the
leaves and 13 lignans from the berries (Table 2) (5,16,17).
1http://www.worldfloraonline.org
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FIGURE 1
Flowchart of Piper cubeba studies inclusion and exclusion criteria.
FIGURE 2
Distribution map of Piper cubeba.
Cubebininolide, hinokinin, yatein, and isoyatein, which
represent the furanofuranic family, are the most known
in Piper species and were identified in the berries, leaves,
and stalks. Ashantin, clusin, cubebin, cubebinone, among
others, were identified from the leaves and berries, while
the hemiarensin was detected only in leaves (5). Yatein
was the most predominant lignan in the berries, more
than cubebin, while hinokinin was the most abundant
lignan in the leaves and the stem from the Indonesia
(5) (Figure 4). Noteworthy, the phytochemical profiling
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FIGURE 3
Representative photos of P. cubeba (A) leaves, (B) flowers, and (C) berries.
FIGURE 4
Structure of the most predominant compounds from P. cubeba.
of the plant was mainly annotated from Indonesian
flora and most of the studies targeted the extraction and
identification of lignans.
Volatile compounds
Altogether, 91 volatile compounds were characterized in
the essential oil, oleoresin, ethanol, and dichloromethane
extracts from P. cubeba. Methyl eugenol (41.31%), eugenol
(33.95%), beta-cubebene (18.3%), and alpha-cubebene (4.1%)
dominated the essential oil while cubebol (26.1%) and beta-
cubebene (12.3%) were the major compounds indentified
from the oleoresin. As for ethanol extract, copaene was the
most dominant compound that represented 13.47% of the
extract followed by napthalene, 1,2,3,5,6,8a-hexahydro with
10.36%. Significant differences were detected between the
extracts (Table 3). For instance, α-cubebene was found to
be 4.1% in the essential oil content, 3.5% in the oleoresin,
and 2.07% in the ethanolic extract; however, it was not
detected at the dichloromethane extract. Copaene, another
example, was detected only at the ethanol extract with
13.47% (18). Propylene glycol dominated the dichloromethane
with 23.82%; however, it was not found in all other
extracts (19). These differences might be attributed to
the different extraction methods, detection techniques as
well as geographic distribution and genetic chemotypes.
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TABLE 1 Vernacular names of Piper cubeba and its distribution.
Vernacular names Distribution References
Hab-ul-Urus, Kabâbah, Kebaba, Hhabb El’arûs Arabic (6,104)
Hendkapeghpegh Armenian (6)
Kabab Chini; Sital Chini Bangladesh (6,104)
Pimenta-Cubeba (Portuguese) Brazil (6)
Gandha Menasu Canada (104)
Biji; Bi Cheng Qie, Cheng Qie, Bi Cheng Qie Chinese (6,104)
Cubebe, Cubebepeper, Staartpeper Dutch
Tailed Peeper English (104)
Cubèbe, Poivre À Queue, Poivre De Java, Poivrier Cubèbe, Quibebes French (6)
Tadamiri Gujrati (104)
Mahilyun, Karifiyun, Koubeba Greek (6,104)
Kabab Chiniha, Kabab Chini (Bengali), Tadamari (Gujerati), Cubab-Chinee, Kabab Chini, Sheetal Chini (Hindu),
Kaba-Chini (Maithili), Vaalmilagu (Malayalam), Kankol (Marathi), Kabachin, Kabab Chini (Oriya), Chinamilagu,
Sinamilagu, Valmilagu (Tamil), Chalavamiriyaalu, Tokamiriyalu (Telugu), Kabab Chini (Urdu); Indonesia:
Kemukus, Temukus (Java), Rinu Katencar, Rinu Caruluk (Sudanese)
India
Cubebe, Pepe A Coda Italian (6)
Kubeba, Kubebu Japanese (6,104)
Cubebe Latin
Valmilaku Malyalam
Kubaba Persian
Cubeba Portuguese, Spanish
Piper De Cubebe Romanian (6)
Sungad-muricha Sanskrit (104)
Tokamiriyalu Telugu
Hind Biberi, Hind Biberi Tohomu, Kebabe, Kebebe, Kebabiye Biber, Kebebiye, Kuyruklu Biber Turkish (6)
Noteworthy, further experiments are required to annotate
the non-volatile constituents of the extracts such as the
ethanol extracts.
Fatty acids and others
Altogether, 19 fatty acids along with their esters were
annotated from P. cubeba berries. They include dodecanoic acid
(lauric acid, 24.05%), hexadecanoic acid (palmitic acid, 11.37%),
9-octadecenoic acid (10.00%), decanoic acid (capric acid,
2.62%), 9,12-octadecadienoic acid (Z,Z) (2.50%), octadecanoic
acid (2.08%), methyl decanoate (capric acid methyl ester,
1.80%), tetradecanoic acid (myristic acid, 1.66%), ethyl-(R,E)-
4-hydroxy-3-methylpent-2-enoate (1.12%), along with other
compounds where their concentrations did not exceed a relative
abundance of 1 like palmitic acid methyl ester (0.65%) and
octanoic acid (caprylic acid) (0.18%) (19). The dichloromethane
fraction gave the presence of two phenolic compounds which
are 4-vinylphenol and 2,4-bis(1,1-dimethylethyl)-Phenol (19).
Besides, several flavonoids and phenolic acids were isolated
from the aqueous extract of P. cubeba fruits such as rutin,
catechin, gallic acid, caffeic acid, syringic acid, ferulic acid
(20,21).
Phenolic, flavonoid and minerals
contents
Determination of total phenolic contents from piper fruit
was investigated using the Folin-Ciocalteu method. It amounted
123.1 and 185.65 µg of GAE/g extract from the ethanolic extract
(20,21), and 1,280 µg of GAE/g extract from the for the
methanolic extract (20,21). The quantification of flavonoids
was carried out as well and amounted 65.83 µg QE/g extract
from in ethanolic extract (20,21). P. cubeba fruit aqueous
extract contained zinc (Zn), selenium (Se), magnesium (Mg),
phosphorus (P), iron (Fe), and manganese (Mn) (22).
Traditional uses of P. cubeba
P. cubeba is extensively used in several ways as powder,
decoction, as an essential oil for numerous purposes. The fruit
is frequently used to treat various diseases such as gastro-tonic
and abdominal pain, anti-asthmatic, and sedative. In Morocco,
the plant has been listed among the medicinal plants used in
cancer treatment. Many of these traditional uses were supported
by scientific evidence. These include antibacterial, nematocidal,
analgesic, and anticancer activities. Moreover, P. cubeba fruits
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TABLE 2 Identified lignans from P. cubeba.
Compound name References
Berries, leaves, and stalks
Cubebininolide (5,13)
Hinokinin
Isoyatein (5)
Yatein (5,16)
Berries and leaves
Ashantin (5)
Clusin (5,13,16)
Cubebin (5,16)
Cubebinone (5)
Dihydrocubebin (5,13)
α-O-Ethylcubebin (5)
β-O-Ethylcubebin
5’-Methoxyhinokinin
2-(30,40-Methylenedioxybenzyl)-3-(30,40-dimethoxybenzyl)-
butyrolactone
Di-O-methyl thujaplicatin methylether
Leaves
Hemiarensin (5)
Berries
Cubebinin (5,16)
(+)-Dihydroclusin (16)
()-Haplomyrfolin
(8R,80R)-4-Hydroxycubebinone (13)
(8R,80R,90S)-5-Methoxyclusin
R-Asarone
R-Methylcubebin
Magnosalin
()-Yatein
2,4,5-trimethoxyphenylacetone
Ethoxyclusin
()-Dihydroclusin
1-(2,4,5-trimethoxyphenyl)-1,2-propanedione
are widely exploited in spice market, and also used as food
preservative, coloring aid and in cosmetics (911).
Biological and pharmacological
activities
Several studies have shown that P. cubeba extracts, essential
oils, and their constituents are endowed with many biological
and pharmacological properties such as antioxidant, anti-
inflammatory, antidiabetic, anticancer, reno-hepatoprotective,
immunomodulatory, antidepressant, antimicrobial, anti-
parasite, insecticidal, wound healing, and antidepressant
activities. However, plant constituents can interact with
biological components in the cells such as proteins and
nucleic acids, toxicity studies are mandatory to ensure the
beneficial and biosafety effects of the tested materials. In
this section, we address the toxicity of P. cubeba, describe
the biological and pharmacological activities of its extracts
and/or compounds, underline some mechanisms of action, and
discuss major findings.
Toxicity studies
Toxicity assessment of P. cubeba extracts were reported in
many studies. For instance, it was shown that, using 3-(4,5-
dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT)
assay, P. cubeba extracts were not toxic to RAW 264.7 cells
(monocyte/macrophage-like cells). In addition, normal oral
fibroblasts treated with P. cubeba based compounds mainly
methylcubebin, dihydrocubebin, and hinokinin showed neither
cytotoxicity signs nor morphological changes (12). In vivo, it
was found that the female Wister rats fed with the methanol
extract of P. cubeba fruits were safe up to a maximum dose
of 2,000 mg/kg body weight. It induced neither changes in
behavioral patterns nor signs and symptoms of toxicity nor
mortality (23). In another study, ()-hinokinin, obtained by
partial synthesis from ()-cubebin isolated from the fruits
of P. cubeba, was orally administered (1 mL/rat) to male
Wistar rats daily for 1 week. This treatment did not cause
any significant weight or water intake changes during the
period of the experiment (24). Elsewhere, male Wistar rats
subjected to treatment by P. cubeba essential oil ranging from
50 to 3,000 mg/kg showed no mortality nor overall behavioral
alteration such as shaking, convulsion, writhing, chewing, pupil
size, feeding behavior, and fecal output (25). The biosafety
status was also monitored using other Piper species such as
P. longum L. fruits which caused no significant acute (24 h)
or chronic (90 days) mortality in mice (26). Likewise, the leaf
extract from P. betle was nontoxic on the glyoxalase system of
Swiss albino mice after 2 weeks of oral administration at 1.5 and
10 mg/kg (27).
Antioxidant activity
Plant constituents are commonly known as the best
source of antioxidants that neutralize reactive oxygen species
(ROS) and free radicals (28). Consequently, an increasing
attention is given to the antioxidant potential of plant-based
molecules and their role in benefiting health and preventing
aging and oxidation-related diseases (29). Like other plants,
the antioxidant activities of P. cubeba extracts and essential
oil were widely evaluated (Table 4). Many in vitro assays
were used, mainly 2,2-diphenyl-1-picrylhydrazil (DPPH) radical
scavenging, ferric-reducing antioxidant power (FRAP), β
carotene bleaching, thiobarbituric acid reactive substances
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TABLE 3 Identified compounds from essential oil, oleoresin, ethanol and dichloromethane extracts of P. cubeba berries.
Compound name Relative abundance (%) References
Essential oil Oleoresin Ethanol extract Dichloromethane
α-Phellandreneb0.2 −−− −−− −−− (20,21)
α-TerpinenebTrace −−− −−− −−− (21)
α-Caryophyllenea1.14 −−− 0.62 −−− (18)
α-Copaeneb8.8 6.2 −−− −−− (18,21)
α-Cubebenea,b4.1 3.5 2.07 −−−
α-Guaieneb0.2 0.2 −−− −−− (18)
α-Gurjuneneb0.3 0.6 −−− −−−
α-Humuleneb0.9 1.5 −−− −−−
α-Muuroleneb0.6 1.3 −−− −−−
α-Pineneb4.1 0.5 −−− −−− (20,21)
α-Selinenea0.47 −−− −−− −−−
α-Terpineola0.96 −−− −−− −−−
α-Terpinolene a1.41, 0.9 −−− −−− −−−
α-Thujene b4.5 0.6 −−− −−−
β-Caryophyllenea,b5.65, 3.7 3.7b−−− −−−
β-caryophyllene oxidea,b0.96, 0.3 −−− −−− −−−
β-Caryophyllene, transa−−− −−− −−− 1.9 (19)
β-Cubebeneb18.3 12.3 −−− −−− (18,21)
β-Elemenea,b0.66, 0.6 1.4b−−− −−− (20,21)
β-Myrcene a,b1.23 a, 0.3 b,−−− −−− −−−
β-Ocimene a0.30 −−− −−− −−−
β-Ocimene, trans −−− 0.2 −−− −−− (21)
β-Phellandreneb5.9 1.2 −−− −−− (20,21)
β-Pineneb0.7 0–2 −−− −−−
γ-Amorpheneb2.0 −−− −−− −−− (18)
δ-3-CarenebTrace −−− −−− −−− (20,21)
δ-Cadinenea,b0.19a, 0.9b2.7 −−− −−− (18,20,21)
δ-Elemene a,b0.9 0–6b−−− −−− (20,21)
δ-Terpineneb0.2 −−− −−− −−− (21)
allo-Aromadendreneb3.1 3.5 −−− −−− (18)
Aromadendreneb0.1 −−− −−− −−−
Bicyclogermacreneb1.5 0.7 −−− −−−
Cadina-1(2),4-diene-transb0.2 1.0 −−− −−−
CamphenebTrace −−− −−− −−− (20,21)
CamphorbTrace −−− −−− −−− (18,21)
Caryophyllene oxidea−−− −−− −−− 0.65 (19)
cis-Sabinene hydrateb0.9 0.3 −−− −−− (18,21)
Citrala−−− −−− −−− 0.6 (19)
Citronellola0.10 −−− −−− −−− (20,21)
Copaene a−−− −−− 13.47 −−− (18)
Cubebolb4.7 26.1 −−− −−−
Cubebol stereoisomerb0.2 5.6 −−− −−−
Cyclohexenea−−− −−− −−− 0.82 (19)
D-Limonenea0.12 −−− −−− −−− (20,21)
Estragolea0.15 −−− −−− −−−
Ethynylbenzenea−−− −−− −−− 0.71 (19)
(Continued)
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TABLE 3 (Continued)
Compound name Relative abundance (%) References
Essential oil Oleoresin Ethanol extract Dichloromethane
Eugenola33.95 −−− −−− −−− (20,21)
Germacrene Da,b0.15a, 2.6b8.3b−−− −−− (18,20,21)
Germacrene-Bb0.1 −−− −−− −−− (18)
Hexanala−−− −−− −−− 0.38 (19)
Isocaryophyllene a−−− −−− 2.23 −−− (18)
Isocembrola0.16 −−− −−− −−− (20,21)
IsoledenebTrace −−− −−− −−− (18,21)
Isopropylpyrazine a−−− −−− 1.52 −−− (18)
Ledol a,b−−− 2.9 6.25 −−− (18,21)
Linaloola,b0.22, 4.9 1.5 b−−− −−− (20,21)
Methyl eugenola41.31 −−− −−− −−− (20,21)
Muurola-4(14), 5-diene trans b−−− 0.4 −−− −−− (18)
Naphthalene, 1,2,3,4,4a,5,6,8a-oct a−−− −−− 1.83 −−−
Naphthalene, 1,2,3,4,4a,7-hexahydro a−−− −−− 2.23 −−−
Napthalene, 1,2,3,5,6,8a-hexahydro a−−− −−− 10.36 −−−
p-Cymeneb1.0 −−− −−− −−− (21)
p-Cymene-8-ol a3.50 −−− −−− −−− (20,21)
Phytol −−− −−− 0.44 (19)
Propylene glycol −−− −−− −−− 23.82 (19)
Sabineneb19.4 5.8 −−− −−− (20,21)
Spathulenola,b0.18, 0.4 −−− −−− −−−
Terpinen-4-ola,b1.80, 0.9 −−− −−− −−−
TerpinolenebTrace −−− −−− −−− (18,21)
trans-Sabinene-hydrateb0.5 0.1 −−− −−− (18,21)
Undecanea−−− −−− −−− 0.31 (19)
Undecanoic acid, ethyl estera−−− −−− 1.36 −−− (18)
Viridiflorola,b0.39, 0.3 −−− −−− −−− (20,21)
Zingibereneb0.1 −−− −−− −−− (18)
(2R,200 R)-(-)-Tetrahydro-2,2-0-biuranyl-5,50-dionea−−− −−− −−− 1.30 (19)
(2S)-3-Methyl-3-buttene-1,2-diol −−− −−− −−− 0.15 (19)
(E)-Geraniola0.19 −−− −−− −−− (20,21)
1,3,6-Heptatriene,2,5,6-trimethyl a−−− −−− 4.54 −−− (18)
1,6-Octadienea−−− −−− 0.25 (19)
1,8-Cineolea,b2.94, trace 0.8 b−−− −−− (20,21)
1H-Cycloprop [e] azulene, decahydro a−−− −−− 3.71 −−− (18)
1-Naphthalenol, decahydro-4a-methyl a−−− −−− 1.37 −−− (18)
2-Pentenea−−− −−− −−− 0.5 (19)
3,7-Dimethyl-2,6-octadienala−−− −−− −−− 1.42
3-Carenea−−− −−− −−− 0.18
3-Cyclohexene-1-methanol −−− −−− −−− 0.22
4-Hydroxy-4-methyl-2-pentanonea−−− −−− −−− 0.29
4-Methylthiazole a−−− −−− 0.18 −−− (18)
5-Undecyne a−−− −−− 1.06 −−− (18)
Trace: <0.05; aGC-MS; bMS and −−− (not found).
(TBARS), phosphomolybdenum, CUPRAC (Cupric reducing
antioxidant capacity), and total antioxidant capacity (TAC)
assays (Table 4).
The antioxidant capacity of six extracts from P. cubeba fruits
(petroleum ether, benzene, ethyl acetate, acetone, methanol,
and ethanol) were evaluated in vitro (18). At 200 µg/mL, it
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TABLE 4 Antioxidant activity of P. cubeba.
Extract/compound Used method Effects References
Dry fruits
Methanol DPPH IC50 = 58.75 µg/mL (50)
Dry berries
n-hexane DPPH % inhibition = 46% (at 10 mg/mL) (105)
Dichloromethane IC50 = 650 µg /mL
MeOH IC50 = 271 µg /mL
Fruits
Essential oil DPPH IC50 = 110.00 ±0.08 µg/mL (106)
FRAP IC50 = 106.00 ±0.11 µg/mL
β-Carotene-linoleate IC50 = 315.00 ±2.08 µg/mL
Methanol DPPH IC50 = 11.3 ±0.3 µg/mL (107)
Essential oil % inhibition = 17.53 ±0.030% (108)
Methanol % inhibition = 66.20 ±3.20% (109)
Essential oil IC50 = 78.9 µg/Ml (8)
Hydro-alcoholic % inhibition = 20.23% (at 500 µg/mL) (110)
IC50, half maximal inhibitory concentration; DPPH, 2,2-diphenyl-1-picryl-hydrazyl-hydrate; FRAP, ferric-reducing antioxidant power.
was noticed that the ethanolic extract was the most potent in
inhibiting DPPH, followed by acetone and ethyl acetate extracts
at the highest concentration tested. Comparable pattern of
antioxidant activities between extracts was observed using other
methods such FRAP and CUPRAC assays. The antioxidant
effect was also reported using P. cubeba essential oil. In fact,
essential oil at 500 µg/mL elicited a good radical scavenging
activity (84%) compared to ascorbic acid (92%), the reference
antioxidant compound (25). Similarly, the radical scavenging
activity induced by P. cubeba essential oil using DPPH and
ABTS assays was up to 38.69% higher than the one that was
exhibited by the essential oil of P. nigrum L. (7). Three other
Piper species, namely P. guineense Schum and Thonn, P. nigrum
L. and P. umbellatum L. showed also endowed scavenging
activity (up to 89.9% inhibition) and metal chelating activity
(up to 93.9% inhibition) (30). Consequently, Piper species can
be considered as a great source of modulators of free radical
induced disorders.
Cytotoxicity and anti-cancer activity
As several cancer chemotherapeutics are derived from
plant-based molecules (31), multiple studies have explored
the antitumor and cytotoxic activity of P. cubeba extracts
against model cancer cells using several methods, mainly
MTT test. For instance, the dichloromethane extract from
P. cubeba fruits induced apoptosis on triple negative breast
cancer cell lines (MCF-7 and MDA-MB-231), colon cancer
cells (HT-29), cholangiocarcinoma cells (KKU-M213), with
less cytotoxicity against normal fibroblast (L929). Sequential
extraction showed that one fraction, named dichloromethane
15 (DE15), significantly enhanced multi-caspases activity in
the breast cancer cell line MDA-MB-231 in a time-dependent
fashion (19). Similarly, Graidist et al. (32) showed that the
methanolic crude extract of P. cubeba fruits exhibited a better
cytotoxic activity against MDA-MB-468 and MCF-7 breast
cancer cell lines than the dichloromethane crude extract. From
the six fractions, the most active fraction had an IC50 of >4
µg/mL. DNA fragmentation assay demonstrated an apoptosis
pattern in MCF-7, MDA-MB-468, MDA-MB-231, and L929
cancer lines, but not in MCF-12A normal cells (32).
Several compounds extracted from P. cubeba have been
explored for their anticancer and cytotoxic potential, among
them (–)-cubebin and its derivatives. Niwa et al. (33) studied
the safety profile of (–)-cubebin by testing its cytotoxicity,
mutagenicity, cell proliferation kinetics, and induction of
apoptosis in human colon adenocarcinoma cells (HT29). MTT
assay showed that (–)-cubebin was cytotoxic at 280 µM,
whereas no cytotoxicity was demonstrated below 28 µM. In
addition, micronucleus assay revealed that (–)-cubebin was
not mutagenic, did not alter cell-growth kinetics over 4 days,
and absence of induced apoptosis after 24 h (33). Moreover,
the effect of P. cubeba extract and its major lignans (cubebin,
dihydrocubebin, ethylcubebin, hinokinin, and methylcubebin)
were evaluated on the larynx (Hep-2) and oral (SCC-25)
squamous carcinoma cells and normal fibroblasts. They all
decreased cell proliferation and migration with no change
in cellular morphology and no genotoxic effects. This was
attributed to the alteration of the expression of genes and
proteins involved in the inflammatory process (12). This
study concluded that cubebin and methylcubebin isolated from
P. cubeba had a great effect on the proliferation, migration,
and genotoxic profile of the head and neck cancer cells. Next
to P. cubeba, the ethanolic extract P. nigrum L. was toxic
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to MCF-7 cells likely through calf thymus DNA (CT DNA)
intercalation and damage (34). Additionally, compounds from
Korean P. kadsura A549 such as kadsuketanone A, piperolactam
A, and piperolactam B elicited a cytotoxic effect toward the
SK-OV-3 (ovarian cancer cells), A549 (non-small cell lung
adenocarcinoma), SK-MEL-2 (skin melanoma), and HCT-15
(colon cancer cells) cell lines (35). Many other Piper plants
were reported to be used traditionally to treat cancer or
cancer-like symptoms. These include P. aduncum L. for skin
tumors (36), P. longum L. for breast cancer (37), P. nigrum
L. for abdominal, respiratory, or gastric tumors/cancers (38
40). A plenty of extracts and compounds from the genus Piper
were proved to be cytotoxic against cancer cells. For instance,
amide alkaloids represent up to 53% of the most bioactive
compounds. Outstandingly, piperlongumine showed excellent
toxicity against dozens of cancer cell lines both in vitro and
in vivo (41). Hence, conducting clinical anticancer studies
on Piper plants, among them P. cubeba, and their bioactive
principles seems to be worthwhile.
Anti-inflammatory activity
Several disorders and diseases are linked to the
inflammatory responses including diabetes mellitus,
rheumatoid arthritis, neurodegenerative diseases, and cancer
(42). Many anti-inflammatory agents were isolated from
plants such as curcumin, quercetin, capsaicin, resveratrol,
and epigallocatechin-3-gallate (43). The anti-inflammatory
activity of P. cubeba was studied in various studies both
in vitro and in vivo. For instance, Mazlan et al. evaluated
the anti-inflammatory effect of P. cubeba extracts and
fractions by monitoring the nitric oxide (NO) production
in lipopolysaccharide (LPS) RAW 264.7 cells. Compared to
untreated cells, those treated with P. cubeba extracts and
fractions showed a significant reduction in NO production
up to 74.17%. The methanolic extract was the most potent in
reducing NO production (44). Similarly, P. cubeba methanolic
extract decreased NO production in macrophage RAW264.7
and HEK293T cells without any evidence of cell toxicity. In
addition, it inhibited the expression level of proinflammatory
cytokines such as iNOS and IL-6, downregulated NF-κB
activation, and reduced the phosphorylation of IκBα, IKKα/β,
Akt, p85, Src, and Syk (45). Interestingly, molecules such as
5-acetyl-2,3-dihydro-7-methyl-1H-pyrrolizine were identified
in P. cubeba fruits aqueous extract and shown to reduce
LPS-induced inflammation and inhibit LDL oxidation (46).
Some mechanisms underlying the inflammatory effect
of P. cubeba, especially its essential oil, were studied using
carrageenan induced pleurisy in rats (25). At 600 mg/kg,
the essential oil substantially reduced the paw edema,
the weight of cotton pellet granuloma, and the exudate
volume. In addition, the level of polymorphonuclear cells
was decreased as well as lung tissue myeloperoxidase, NO,
and proinflammatory cytokines such as TNFαand IL-
1β. The anti-inflammatory effects observed in vivo were
attributed to the fact that P. cubeba essential oil contains
sabinene, γ-terpinene, 4-terpineol, and α-thujene, known to be
endowed with antioxidant and anti-inflammatory properties.
Comparatively, the bioactive n-hexane and methanolic
extracts from P. kadsura aerial parts were found to contain
kadsuketanone A, ent-germacra-4(15),5,10(14)-trien-1β-ol,
aristolactam A II, trans-2,3-diacetoxy-1-[(benzoy1oxy)methyl]-
cyclohexa-4,6-diene, and piperolactam A and B. Both extracts
induced a significant inhibition of both PGE2and NO
production in the LPS-activated microglia cells (35). Thus,
expanding the research on the anti-inflammatory potential of
Piper species can be a promising strategy to develop Piper-
derived drugs or adjuvant medicines suitable for the treatment
of inflammation-related disorders.
Antidiabetic activity
Diabetes is one of the most prevalent health problems
worldwide (47). It has serious health consequences leading
to increasing mortality. Synthetic anti-diabetic drugs have
unavoidable side effects. Therefore, medicinal plants and their
active components can act as alternative anti-diabetic medicines.
Many plants are renowned for antidiabetic potential including
P. cubeba (4749). Noteworthy, the role of P.cubeba in the
management of diabetes has been underexploited and is yet to
receive sound scientific interest.
Muchandi et al. (50) demonstrated that the ethanolic extract
of P. cubeba fruits administered to Albino rats protected
them against D-galactose induced neuronal lipofuscinogenesis
(51). In fact, using a dose of 400 mg/kg, p.o., of P. cubeba
fruits significantly reduced lipofuscin fluorescence from the
hippocampus region of animals comparatively to D-galactose
treated rats. In addition, a decrease in the accumulation of
lipofuscin granules in hippocampus of animals’ brains was
observed in P. cubeba treated group. Observed effect was
suggested to be due to the richness of the extract in lignans,
mainly cubebin, hinokinin, yatein, and isoyatein, that are known
as strong antioxidants.
As the intestinal enzymes α-amylase and α-glucosidase are
key targets in the regulation of diabetes mellitus, P. cubeba
extracts were reported as digestive enzymes inhibitors. It was
found that the methanolic and aqueous extracts at 1 mg/mL
were able to significantly inhibit α-glucosidase and α-amylase
in vitro (48). Moreover, the anti-diabetes activity of both
extracts has been suggested to be likely associated with their
antioxidant properties. Next to P. cubeba, the root aqueous
extract of P. longum administered by intraperitoneal route
to streptozotocin induced diabetic male Wister albino rats at
200 mg/kg, b.w for 30 days, decreased, by 66.7%, the fasting
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blood glucose. These studies justify the traditional use of Piper
species, including P. cubeba, and open up promising avenues for
the management of diabetes and related complications.
Antimicrobial activity
Antibacterial activity
Nowadays, the increasing prevalence of antibiotic resistance
is one of the major challenges for public health worldwide.
It has been attributed to the over- and misuse of antibiotics,
as well as a declining trend in novel drug development
by the pharmaceutical industry and challenging regulatory
requirements (52,53). As plants represent a great resource in
drug discovery, for being mostly biocompatible, biodegradable,
and less cytotoxic, their extracts and secondary metabolites are
being widely explored to discover potential next antimicrobials
(29,54).
Extracts and compounds from P. cubeba parts, mainly fruits,
were largely evaluated for antimicrobial activity (Table 5). Using
four extracts (acetone, hexane, methanol, and ethanol) from
P. cubeba fruits, Akshita et al. (55) found that all extracts
showed high to moderate antibacterial activity against Klebsiella
sp., Staphylococcus aureus,Escherichia coli,Enterococcus sp.,
Enterobacter sp., and Pseudomonas aeruginosa except hexane
extract which exhibited no activity (Table 5). The best effects
were observed toward Enterococcus sp. followed by E. coli and
P. aeruginosa (55). Similar observation was reported that the
hexane extract was not active in inhibiting different microbial
strains (56). Noteworthy, P. cubeba extracts were more effective
on Gram-positive than against Gram-negative bacteria. This
is due to the single-layer cell wall in Gram-positive strains in
contrast to the multilayered cell wall of Gram-negative bacteria
that constitutes a barrier for the invasion of antimicrobial agents
through the cell membrane.
Moreover, P. cubeba essential oil was also shown to be
endowed with good antibacterial activity against methicillin-
resistant S. aureus ATCC 43300 (Table 5). This was evaluated
using atomic force microscopy and transmission electron
microscopy. At 50 µg/mL, the essential oil severely damaged the
bacterial cell walls while it was not active at microscopic levels at
25 µg/mL. However, at nanoscopic levels, it induced significant
perturbation in the bacterial cell wall. These effects on the cell
wall and plasma (cytoplasmic) membrane are likely to be the
way by which this essential oil impaired bacterial activity (57).
Elsewhere, P. cubeba essential oil induced anti-Helicobacter
pylori activity (MIC = 7.81 µg/mL) and thereby proposed as a
therapeutic agent to protect and/or treat H. pylori infection (58).
P. cubeba methanolic extract was also tested as a natural food
preservative against microbial population in tofu using total
plate counting (TPC) method. A decrease of upper to 3 Log10
CFU/g of TPC was observed against B. cereus, coliform and
E. coli in tofu treated with 0.5% of the extract for 4 h. P. cubeba L.
berries were suitable for use as a natural preservative to reduce
the microbial load in raw food (59).
It was suggested that the essential oil targets the cell
wall of bacterial cells, whereas the extracts attack and destroy
the peptidoglycan causing cell collapse. P. cubeba essential
oil injures the cell wallanchored proteins that are involved
in biofilm formation and adhesion. It could also injure the
cytoplasmic membrane (57). Other Piper plants were reported
for their huge antibacterial spectrum (60). For instance, Piper
nigrum L. methanolic and chloroform extracts inhibited E. coli,
S. aureus,S. typhi, and Proteus sp. except P. aeruginosa
which was resistant to both extracts (61). Likewise, the
leaf ethanolic extract of Piper betel L. exhibited pronounced
antibacterial activities toward B. subtilis,S. aureus,E. coli,
and moderate inhibition of P. aeruginosa. The aqueous extract
was also tested and was active only against B. subitilis
(62). These studies corroborate the traditional use of Piper
plants, including P. cubeba, in managing infectious diseases.
Therefore, it could serve as a source of novel therapeutic
agents against human pathogenic bacteria and food borne
pathogens. Alqadeeri et al. (63) isolated and identified, for the
first time, two compounds, β-asarone, and asaronaldehyde, from
the methanolic extract of P. cubeba and its fractions. Both
compounds inhibited the growth of B. pumilus ATCC14884, B.
cereus ATCC33019, B. megaterium ATCC14581, and B. subtilis
ATCC6633 (MIC = 63.0–125.0 µg/mL) and inactivated more
than 90.99% of the Bacillus spores 0.05%. More importantly,
the compounds destroyed all the spores at 0.1% after 1 h
of incubation. The antibacterial and anti-sporicidal activity
of β-asarone and asaronaldehyde provide useful information
about the antimicrobial effect of P. cubeba (63). However,
in vivo studies would be of a great importance to support their
development as antibacterial agents.
Antifungal activity
The antifungal activity of P. cubeba was fully explored
in many studies by using several methods mainly agar disc
diffusion, well diffusion method, microdilution, inverted petri
plate, and poison food medium assays (Table 6). The antifungal
potential of five extracts of P. cubeba berries against the
opportunistic oral fungal pathogens Candida albicans and
Saccharomyces cerevisiae was studied using the MIC assay (64).
The acetone extract was the most potent against both species
followed by the methanolic and ethanolic extracts. C. albicans
was more sensitive than S. cerevisiae (64). Similarly, Salkar
et al. (65) developed an oral gel from the essential oil (0.5%)
of P. cubeba and tested its activity toward different strains
of Candida. The gel elicited excellent activity against both
normal (C. albicans ATCC 10231, C. glabrata H04FS fluconazole
sensitive, C. krusei G06FS fluconazole sensitive) and resistant
(C. albicans-fluconazole resistant, C. krusei G03FR fluconazole
resistant, C. glabrata H05FR fluconazole resistant) Candida
species. Interestingly, the developed oral gel was endowed
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TABLE 5 Antibacterial activity of P. cubeba.
Bacteria Extract Used method Effects References
E. coli Steam-distilled oil Disk diffusion method ZI = 9 mm (111,112)
B. subtilis ZI = 11 mm
V. cholerae ZI = 11 mm
S. aureus ZI = 9 mm
S. albus ZI = 10 mm
S. dysenteriae ZI = 10 mm
C. dyphteriae ZI = 13 mm
S. typhi ZI = 12 mm
S. lutea ZI = 9 mm
S. faecalis ZI = 11 mm
B. pumtlus ZI = 16 mm
S. pyogenes ZI = 14 mm
M. luteus ZI = 8 mm
P. solanacearum ZI = 30 mm
E. coli Acetone Disk diffusion method ZI = 10 mm; MIC = 0.5 mg/L (113)
S. aureus ZI = 16 mm; MIC = 1 mg/L
P. aeruginosa ZI = 13 mm; MIC = 0.5 mg/L
E. coli Chloroform ZI = 12 mm; MIC = 0.5 mg/L
S. aureus ZI = 11 mm; MIC = 1 mg/L
P. aeruginosa ZI = 10 mm; MIC = 0.5 mg/L
E. coli Ethanolic ZI = 10 mm; MIC = 1 mg/L
S. aureus ZI = 15 mm; MIC = 1 mg/L
P. aeruginosa ZI = 13 mm; MIC = 0.5 mg/L
E. coli Aqueous ZI = 15 mm; MIC = 1 mg/L
S. aureus ZI = 8 mm; MIC = 1 mg/L
P. aeruginosa ZI = 15 mm; MIC = 0.5 mg/L
B. subtilis Essential oil Disk diffusion method ZI = 17 mm (114)
E. coli (ATCC43895) ethanolic extract Microdilution method MIC = 0.63 mg/mL; MBC = 1.25 mg/mL (115)
B. cereus (ATCC 11778) n-hexane Disk diffusion method ZI = 12 mm (105)
Dichloromethane ZI = 23 mm
MeOH ZI = 11 mm
S. aureus (NCTC 1803) n-hexane ZI = 17 mm
Dichloromethane ZI = 16 mm
S. mutans Acetone Agar well diffusion method ZI = 12.64 mm; MIC = 50 mg/mL (64)
Methanol ZI = 12.31 mm; MIC = 50 mg/mL
Ethanol ZI = 13 mm; MIC = 50 mg/mL
S. aureus Acetone ZI = 18.96 mm; MIC = 25 mg/mL
Methanol ZI = 17.65 mm; MIC = 25 mg/mL
Ethanol ZI = 17.32 ±0.57 mm; MIC = 25 mg/mL
B. cereus (JN 934390) Essential oil Agar well diffusion method
and microdilution method
ZI = 15.0 mm; MIC = 3.12 mg/mL;
MBC = 12.5 mg/mL
(20)
B. subtilis (JN 934392) ZI = 16.0 ±0.7 mm; MIC = 6.25 mg/mL;
MBC = 12.5 mg/mL
S. aureus (ATCC 6538) ZI = 19.5 mm; MIC = 1.56 mg/mL;
MBC = 3.12 mg/mL
L. monocytogenes (ATCC 19115) ZI = 19.0 mm; MIC = 1.56 mg/mL;
MBC = 3.12 mg/mL
M. luteus (NCIMB 8166) ZI = 16.0 mm; MIC = 1.56 mg/mL;
MBC = 6.25 mg/mL
(Continued)
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TABLE 5 (Continued)
Bacteria Extract Used method Effects References
K. pneumoniae (ATCC 10031) ZI = 13.0 mm; MIC = 3.12 mg/mL;
MBC = 6.25 mg/mL
S. enterica (ATCC 43972) ZI = 23.0 mm; MIC = 12.5 mg/mL;
MBC = 25 mg/mL
S. typhimurium (ATCC 19430) ZI = 13.5 mm; MIC = 6.25 mg/mL;
MBC = 12.5 mg/mL
E. coli (ATCC 25922) ZI = 21.0 mm; MIC = 3.12 mg/mL;
MBC = 6.25 mg/mL
P. agglomerans Essential oil Agar well diffusion method ZI = 5 mm (116)
X. campestris pv. citri ZI = 11 mm
S. aureus MTCC 3103 Diethyl ether oleoresin Agar well diffusion method 2 µL/well: ZI = 22.mm (117)
6µL/well: ZI = 36.1 mm
B. subtilis MTCC 1790 2 µL/well: ZI = 0 mm
6µL/well: ZI = 13.2 mm
E. coli MTCC 1672 2 µL/well: ZI = 29.2 mm
6µL/well: ZI = 53.3 mm
S. typhi MTCC 733 2 µL/well: ZI = 47.2 mm
6µL/well: ZI = 65.2 mm
S. aureus MTCC 3103 Ethanol oleoresin 2 µL/well: ZI = 12.4 mm
6µL/well: ZI = 18.3 mm
B. subtilis MTCC 1790 2 µL/well: ZI = 18.1 mm
6µL/well: ZI = 32.0 mm
E. coli MTCC 1672 2 µL/well: ZI = 14.3 mm
6µL/well: ZI = 29.3 mm
S. typhi MTCC 733 2 µL/well: ZI = 56.3 mm
6µL/well: ZI = 100 mm
S. aureus MTCC 3103 Petroleum ether, benzene
oleoresin
2µL/well: ZI = 23.6 mm
6µL/well: ZI = 42.1 mm
B. subtilis MTCC 1790 2 µL/well: ZI = 36.3 mm
6µL/well: ZI = 73.3 mm
E. coli MTCC 1672 2 µL/well: ZI = 0 mm
6µL/well: ZI = 21.4 mm
S. typhi MTCC 733 2 µL/well: ZI = 56.4 mm
6µL/well: ZI = 60.0 mm
S. aureus MTCC 3103 Chloroform oleoresin 2 µL/well: ZI = 17.1 mm
6µL/well: ZI = 26.4 mm
B. subtilis MTCC 1790 2 µL/well: ZI = 23.5 mm
6µL/well: ZI = 44.9 mm
E. coli MTCC 1672 2 µL/well: ZI = 50.0 mm
6µL/well: ZI = 68.3 mm
S. typhi MTCC 733 2 µL/well: ZI = 49.2 mm
6µL/well: ZI = 84.1 mm
S. aureus MTCC 3103 Methanol oleoresin 2 µL/well: ZI = 19.2 mm
6µL/well: ZI = 30.2 mm
B. subtilis MTCC 1790 2 µL/well: ZI = 0 mm
6µL/well: ZI = 0 mm
E. coli MTCC 1672 2 µL/well: ZI = 28.5 ±1.4 mm
6µL/well: ZI = 67.0 ±0.1 mm
(Continued)
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TABLE 5 (Continued)
Bacteria Extract Used method Effects References
S. typhi MTCC 733 2 µL/well: ZI = 32.6 ±0.6 mm
6µL/well: ZI = 41.3 ±2.1 mm
S. aureus MTCC 3103 Essential oil 2 µL/well: ZI = 30.6 ±0.3 mm
6µL/well: ZI = 50.4 ±1.6 mm
B. subtilis MTCC 1790 2 µL/well: ZI = 46.6 ±1.2 mm
6µL/well: ZI = 72.3 ±1.1 mm
E. coli MTCC 1672 2 µL/well: ZI = 42.0 ±0.3 mm
6µL/well: ZI = 80.0 ±0.3 mm
S. typhi MTCC 733 2 µL/well: ZI = 56.3 ±0.1 mm
6µL/well: ZI = 100
E. coli Hydro-alcoholic Disk diffusion method ZI = 18 ±0.64 mm (117)
S. saprophyticus ZI=19±0.26 mm
K. pneumonia ZI=19±0.51 mm
P. mirabilis ZI=20±0.41 mm
E. coli Agar Well method ZI = 20 ±0.00 mm
S. saprophyticus ZI=18±0.47 mm
K. pneumonia ZI=18±0.18 mm
P. mirabilis ZI=19±0.37 mm
E. coli n-Hexane Agar well diffusion method ZI = 30 ±2.3; MIC = 7.5 mg/mL (117)
Salmonella sp. ZI = 32 ±2.0; MIC = 5.0 mg/mL
S. flexneri ZI=37±1.6; MIC = 5.0 mg/mL
V. parahaemolyticus ZI=38±0.6; MIC = 5.0 mg/mL
V. cholerae ZI=31±2.0; MIC = 5.0 mg/mL
P. aeruginosa ZI=17±0.6; MIC = 5.0 mg/mL
L. delbrueckii ZI=25±0.7; MIC = 10.0 mg/mL
Brochothrix sp. ZI = 11 ±0.6; MIC = 20.0 mg/mL
B. subtilis Essential oil Agar well diffusion method ZI = 26.5 ±1.1 mm (117)
S. aureus ZI = 41.2 ±1.5 mm
B. cereus Total inhibition
E. coli ZI = 25.6 ±1.5 mm
S. typhi ZI = 17.0 ±0.2 mm
P. aeruginosa ZI = 0 mm
B. subtilis Tailed pepper oleoresin Agar well diffusion method ZI = 32.1 ±0.8 mm
S. aureus Total inhibition
B. cereus ZI = 53.2 ±1.5 mm
E. coli ZI = 19.4 ±1.2 mm
S. typhi ZI = 33.2 ±1.1 mm
S. aureus Acetone Agar well diffusion method ZI = 14.0 ±0.70 mm
Klebsiella sp. ZI = 14.3 ±0.18 mm
Enterococcus sp. ZI = 15.2 ±0.52 mm
P. aeruginosa ZI = 15.3 ±0.62 mm
E. coli ZI = 16.3 ±0.75 mm
S. aureus Methanol Agar well diffusion method ZI = 11.5 ±0.30 mm
Enterococcus sp. ZI = 17.6 ±0.80 mm
P. aeruginosa ZI = 13.2 ±0.06 mm
E. coli ZI = 15.0 ±0.30 mm
S. aureus Ethanol Agar well diffusion method ZI = 9.0 ±0.05 mm
Klebsiella sp. ZI = 8.5 ±0.10 mm
(Continued)
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TABLE 5 (Continued)
Bacteria Extract Used method Effects References
Enterococcus sp. ZI = 11.3 ±0.16 mm
P. aeruginosa ZI = 9.6 ±0.34 mm
E. coli ZI = 8.5 ±0.17 mm
S. mutans KCCM3309 Methanol (100 mg/mL) Disc Diffusion Assay and
microdilution method
ZI = 10.00 mm; MIC = 0.23 mg/mL;
MBC = 0.23 mg/mL
(117)
Ethanol (100 mg/mL) ZI = 10.33 mm; MIC = 0.10 mg/mL;
MBC = 0.10
Hexane (100 mg/mL) ZI = 10.00 mm; MIC = 0.10 mg/mL;
MBC = 0.10
S. sobrinus ATCC33478 Methanol (100 mg/mL) ZI = 12.17 mm; MIC = 0.93 mg/mL;
MBC = 16.67 mg/mL
Ethanol (100 mg/mL) ZI = 11.13 mm; MIC = 0.80 mg/mL;
MBC = 20.83 mg/mL
Hexane (100 mg/mL) ZI = 12.07 mm; MIC = 0.87 mg/mL;
MBC = 20.83 mg/mL
A. viscosus ATCC15987 Methanol (100 mg/mL) ZI = 11.67 mm; MIC = 0.80 mg/mL;
MBC = 20.83 mg/mL
Ethanol (100 mg/mL) ZI = 11.17 mm; MIC = 0.47 mg/mL;
MBC = 20.83 mg/mL
Hexane (100 mg/mL) ZI = 12.67 ±0.58 mm;
MIC = 0.87 ±0.70 mg/mL;
MBC = 13.54 ±10.97 mg/mL
S. mutans KCCM3309 Hexane fraction
(100 mg/mL)
ZI = 9.33 mm; MIC = 0.10 mg/mL;
MBC = 0.10 mg/mL
Ethyl acetate fraction
(100 mg/mL)
ZI = 9.66 mm; MIC = 0.13 mg/mL;
MBC = 0.13
Aqueous methanol fraction
(100 mg/mL)
ZI = 11.00 mm; MIC = 0.10 mg/mL;
MBC = 0.10
S. sobrinus ATCC33478 Hexane fraction
(100 mg/mL)
ZI = 11.13 mm; MIC = 0.67 mg/mL;
MBC = 25.00 mg/mL
Ethyl acetate fraction
(100 mg/mL)
ZI = 11.07 mm; MIC = 3.15 mg/mL;
MBC = 25.00 mg/mL
Aqueous methanol fraction
(100 mg/mL)
ZI = 11.20 mm; MIC = 1.07 mg/mL;
MBC = 7.29 mg/mL
Ac. viscosus ATCC15987 Hexane fraction
(100 mg/mL)
ZI = 12.00 mm; MIC = 1.58 mg/mL;
MBC = 16.67 mg/mL
Ethyl acetate fraction
(100 mg/mL)
ZI = 10.83 mm; MIC = 1.20 mg/mL;
MBC = 25.00 mg/mL
Aqueous methanol fraction
(100 mg/mL)
ZI = 12.33 mm; MIC = 1.07 µg/mL;
MBC = 12.50 µg/mL
IZ, inhibition zone; MBC, minimum bactericidal concentration; MIC, minimum inhibitory concentration.
with comparable inhibitory effect to marketed local-herbal gel
samples (65). Hence, the crude extracts as well as essential oil
from P. cubeba fruits were considered as promising treatments
of oral fungal infections, especially C. albicans species.
Many oleoresins from P. cubeba fruits were tested against
different food pathogenic fungi (66) (Table 6). Using inverted
petri plate assay, the chloroform oleoresin at 6 µL was
highly active against Penicillium purpurogenum. However,
the petroleum benzene oleoresin was ineffective against
Fusarium oxysporum at all doses. Other oleoresins elicited
minimum to moderate activities. Using the food poison
technique, the ethanol oleoresin at 6 µL was effective against
Penicillium madriti. Many other fungal species were sensitive
to P. cubeba essential oil such as Aspergillus fumigatus,
A. flavus, and F. solani, among others. The antifungal activities
of P. cubeba extracts and essential oils are presented in
Table 6.
Comparatively, the crude methanolic extract and
fractions (dichloromethane, hexane, and ethyl acetate)
from Piper solmsianum, as well as four pure compounds
namely eupomatenoid-5, eupomatenoid-3, conocarpan
and orientin were all assessed against 12 pathogenic fungi
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TABLE 6 Antifungal activity of P. cubeba.
Fungi Extract/compound Used method Effects References
G. lucidum Essential oil Disk diffusion method ZI = 10 mm (114)
C. albicans Acetone extract Agar well ZI = 15.31 ±0.57 mm; MIC = 12.5 mg/mL (64)
Methanol extract diffusion ZI = 12.31 ±0.57 mm; MIC = 25 mg/mL
Ethanol extract method ZI = 11.94 ±1 mm; MIC = 25 mg/mL
S. cerevisiae Acetone extract ZI = 10.93 ±1 mm; MIC = 12.5 mg/mL
Methanol extract ZI = 11.31 ±0.57 mm; MIC = 12.5 mg/mL
Ethanol extract ZI = 11.64 ±0.57 mm; MIC = 12.5 mg/mL
Pythium catenulatum (AY598675) Essential oil Microdilution method ZI = 13.0 ±0.5 mm; MIC = 6.25 mg/mL;
MFC = 25 mg/mL
(20)
Fusarium oxysporum (AB586994) ZI = 17.0 ±0.1 mm; MIC = 3.12 mg/mL;
MFC = 12.5 mg/mL
Fusarium sp. (JX391934) ZI = 15.0 ±0.8 mm; MIC = 3.12 mg/mL;
MFC = 12.5 mg/mL
C. albicans ATCC 10231 Essential oil at 0.5 % Agar dilution method MIC = 50 mg/mL
Candida albicans-fluconazole resistant MIC = 50 mg/mL
Candida krusei G03FR fluconazole resistant MIC = 60 mg/mL
Candida krusei G06FS fluconazole sensitive MIC = 60 mg/mL
Candida glabrata H04FS fluconazole sensitive MIC = 50 mg/mL
Candida glabrata H05FR fluconazole resistant MIC = 60 mg/mL
G. candidum (TMa 001) Ethanol extract of berries Disc diffusion method ZI = 7.26 ±0.20 mm
P. citrinum (GRd 001) ZI = 7.13 ±0.20 mm
T. hirsuta (LMd 001) ZI = 13.80 ±1.40 mm
G. candidum (TMa 001) Methanol extract of
berries
ZI = 8.10 ±0.80 mm; MIC = 1.25 mg/mL;
MFC = 2.5 mg/mL
P. citrinum (GRd 001) ZI = 7.67 ±0.90 mm;
MIC = 0.625 mg/mL; MFC = 1.25 mg/mL
T. hirsuta (LMd 001) ZI = 18.30 ±3.00 mm;
MIC = 0.039 mg/mL; MFC = 0.078 mg/mL
G. candidum (TMa 001) Methanol extract of
berries
Conidial germination = 3.1%
P. citrinum (GRd 001) Conidial germination = 10.0%
T. hirsuta (LMd 001) Conidial germination = 21.6%
Penicillium purpurogenum (MTCC 1786) Diethyl ether Inverted petri plate
method
2µL: ZI = 28 mm
6µL: ZI = 52 mm
Fusarium oxysporum (MTCC 284) 2 µL: ZI = 18 mm
6µL: ZI = 21 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 36.2 mm
6µL: ZI = 45 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 31.3 mm
6µL: ZI = 35 mm
Penicillium purpurogenum (MTCC 1786) Ethanol 2 µL: ZI = 13 mm
6µL: ZI = 36 mm
Fusarium oxysporum (MTCC 284) 2 µL: ZI = 10 mm
6µL: ZI = 14 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 60 mm
6µL: ZI = 75 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 62.5 mm
6µL: ZI = 65 mm
(Continued)
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TABLE 6 (Continued)
Fungi Extract/compound Used method Effects References
Penicillium purpurogenum (MTCC 1786) Petroleum benzene 2 µL: ZI = 35 mm
6µL: ZI = 69 mm
Fusarium oxysporum (MTCC 284) 2 µL: ZI = 0 mm
6µL: ZI = 2 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 32 mm
6µL: ZI = 40 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 37.5 mm
6µL: ZI = 42 mm
Penicillium purpurogenum (MTCC 1786) Chloroform 2 µL: ZI = 48 mm
6µL: ZI = 83 mm
Fusarium oxysporum (MTCC 284) 2 µL: ZI = 15 mm
6µL: ZI = 22 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 13 mm
6µL: ZI = 40 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 42 mm
6µL: ZI = 53 mm
Penicillium purpurogenum (MTCC 1786) Methanol 2 µL: ZI = 30 mm
6µL: ZI = 61 mm
Fusarium oxysporum (MTCC 284) 2 µL: ZI = 3.7 mm
6µL: ZI = 10 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 8.7 mm
6µL: ZI = 25 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 56 mm
6µL: ZI = 62 mm
Penicillium purpurogenum (MTCC 1786) Essential oil 2 µL: ZI = 90 mm
6µL: ZI = 100 mm
Fusarium oxysporum (MTCC 284) 2 µL: ZI = 12.5 mm
6µL: ZI = 35 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 10 mm
6µL: ZI = 12.5 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 65 mm
6µL: ZI = 73 mm
Fusarium proliferatum (MTCC 2935) 2 µL: ZI = 52 mm
6µL: ZI = 62.3 mm
Penicillium madriti (MTCC 3003) 2 µL: ZI = 41.7 mm
6µL: ZI = 70 mm
Trichophyton rubrume Fruit 70% Ethanol extract Microdilution method MIC = 8 mg/mL
Fruit hot-water extract MIC >8 mg/mL
Terbinafine MIC = 2 µg/mL
Alternaria porri Essential oil Disc diffusion method IZ = 7 ±1 mm
Fusarium oxysporum f. sp cicer IZ = 7.5 ±1.29 mm
IZ, inhibition zone; MBC, minimum bactericidal concentration; MIC, minimum inhibitory concentration.
(67). The methanolic extract and fractions elicited a good
antifungal effect against all the dermatophytes strains (MIC,
µg/mL = 20–60), a weak activity against the zigomycetes
and were not active toward the hyaline hyphomycetes.
Compounds eupomatenoid-5, conocarpan, and orientin
exhibited pronounced activities against all the dermatophytes
tested (MIC 1–9 µg/mL). Noteworthy, conocarpan showed
a remarkable activity against all the yeasts. To sum up,
the antifungal activity of P. cubeba and its relatives seems
to be promising and is likely related to the presence of
bioactive compounds belonging to neolignanes and flavonoids.
However, the presence of other active compounds should be
verified and evaluated.
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Antiparasitic and antileishmanial
activities
In addition to the antimicrobial activities, P. cubeba
essential oil was also active against Schistosoma mansoni, the
trypomastigote and amastigote forms of Trypanosoma cruzi, and
the promastigote forms of Leishmania amazonensis (68). The
in vitro inhibitory effect against T. cruzi was dose dependent.
In contrast, essential oil was inactive toward L. amazonensis.
In vivo, a recent study showed that intraperitoneal treatment
of male BALB/c mice by encapsulated and unencapsulated ()-
cubebin isolated from P. cubeba showed up to 61.3% reduction
in the number of the trypomastigotes of a strain of T. cruzi.
Animals treated with encapsulated ()-cubebin survived longer
compared to those treated with Benznidazole used as standard
antiparasitic drug (69). These findings open a promising
application of encapsulated ()-cubebin as antiparasitic agent.
Other Piper species were also reported for antiparasitic
purposes. For instance, P. dennisii was shown to exhibit
anti-plasmodial activity in vitro (70). Moreover, benzoic acid
derivatives isolated from P. acutifolia and P. glabratum were
effective toward both T. cruzi and Plasmodium falciparum (71).
In vitro evaluation of extracts from different Piper plants such
as P. barbatum,P. aduncum,P. acutifolium, and P. dilatatum
showed that they are potent in inhibiting T. cruzi (72). These
findings demonstrate that the antiparasitic potential of Piper
plants, including P. cubeba, is worth exploring in drug discovery.
Antileishmanial activity of P. cubeba extracts was evaluated
in vitro toward Leishmania donovani promastigotes (73). All
tested extracts (n-hexane, ethyl acetate, methanol, and acetone)
elicited a significant activity at 100 µg/mL with more than 90%
inhibition. In the case of n-hexane extract, two lignans namely
cubebin and hinokinin, were identified and isolated. Cubebin
exhibited a significant in vitro antileishmanial activity at 100
µM. In vivo experiment carried out in golden hamsters against
L. donovani amastigotes showed that cubebin slightly reduced
parasitic burden and spleen weight (73). Comparatively,
the antileishmanial activity was also demonstrated by of
P. aduncum extracts (74). Moreover, the essential oils from
P. angustifolium were effective against Leishmania infantum
(75). Also P. cubeba exhibited anthelmintic activity against
earthworms and tapeworms in vitro (76).
Wound-healing activity
Medicinal plants are the major source of wound healing
products with more than 70% while the remaining sources
are mineral and animal-based pharma products (7779).
Several plants are known to accelerate wound healing (80).
However, only few studies have explored this activity from
P. cubeba. Shakeel et al. (81) assessed the wound healing
effect of P. cubeba essential oil using self-nanoemulsifying
drug delivery system (SNEDDS). Prepared formulation was
evaluated for wound healing, collagen determination, and
histo-morphological examination in female Wistar rats. Upon
oral administration, it was found that EO-SNEDDS formula
significantly accelerated wound healing and enhanced collagen
content in tested animals in comparison with pure essential oil.
Noteworthy, histopathological evaluation of the formula-treated
animals showed no signs of inflammatory cells indicating that it
is safe to female rats (81).
More recently, essential oil from P. cubeba fruits (PCEO)
was tested for in vivo wound healing potential (20). Tested
PCEO induced a powerful antibacterial activity especially
against Listeria monocytogenes and S. aureus, known to be
involved in wound infections. Interestingly, the application of
PCEO as topical cream accelerated the wound healing process,
increased the SOD level, and reduced the malondialdehyde
(MDA) level. In addition, histopathological examination
demonstrated that the derma was restored and arranged
properly. The observed activities were attributed to the synergy
between the antioxidants and antimicrobials present in PCEO.
Phytochemicals seem to elicit wound healing activity by
targeting several factors mainly those known to be responsible
for delaying and/or reducing the wound healing process such as
infections, deficiency in blood supply, diabetes mellitus, necrotic
tissue, and lymphatic blockage (82). Within Piper genus, the
aqueous leaf extract of P. betle applied to wounds in vivo
induced a significant contraction and complete epithelization
of the wounds after 10 and 14 days of treatment, respectively
(83). Elsewhere, topical application of the ointments prepared
from the leaves, stems, and roots of P. hayneanum significantly
improved the healing of rats’ wounds and reduced the infections
by two wound pathogens: S. aureus and C. albicans (84).
In conclusion, it is apparent that Piper plants contain active
principles with great potential to be used as topical ointments
to enhance wound healing and prevent the establishment of
wound-related infections.
Immunomodulatory activity
Phytochemicals, such as terpenoids, polysaccharides,
glucosides, flavonoids, and alkaloids, are widely reported as
immunomodulators to some extent (85). As for P. cubeba,
the immunomodulatory activity of its protein extracts was
evaluated on the proliferation of immune cells using MTT assay
on the splenocytes. This was tested in presence and absence of
the mitogenic agent, concanavalin-A (Con-A) (86). The protein
extracts exhibited a more significant immunosuppressive
activity compared to the total extract. In addition, Ikawati et al.
(87) demonstrated that the hexane and ethanolic extracts of
P. cubeba fruits caused lysis of 2H3 cells leading to the release
of high level of histamine (87). This effect was comparable
to that induced by the standard drug, Thapsigargin. These
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results suggest the potential of P. cubeba extracts to face
allergic diseases. However, in vivo experiments would provide
further useful information to identify the bioactive molecules
and explain the underpinning mechanisms. Using another
Piper member, the administration of the methanolic extract of
P. longum and its major principle piperine, induced a significant
increase in the total white blood cell (WBC) count, enhanced
the bone marrow cellularity, and increased circulating antibody
titer, α-esterase positive and plaque forming cells in Balb/c
mice (88).
Several other phytochemicals have been evaluated for
immunomodulatory purposes and some mechanisms of action
have been uncovered. For instance, epigallocatechin-3-gallate
(EGCG) was able to inhibit NF-kB activation and down-
regulate the production of NO in macrophages as well as the
expression of monocyte chemoattractant protein-1 (MCP-1).
Moreover, resveratrol, the highest renowned active molecule in
grapevine, acted via inhibiting TNF-αand/or (LPS)-mediated
macrophages, NF-kB, dendritic cells, and myeloid (89).
Hepato- and renoprotective activity
As the need for anti-hepatitis C virus (HCV) agents is
growing, the search for new candidates that can serve as drugs
or as core-entities to design an effective HCV inhibitor and its
enzymes is promising. P. cubeba aqueous extract inhibited HCV-
PR activity in vitro with an IC50 of 18.0 µg/mL (45). When
compared to other plants tested in this study (45), P. cubeba
aqueous and methanol extracts were among the most active by
inducing 94.2 ±2.1 and 84.7 ±1.8% inhibition at 100 mg/mL,
respectively. In an attempt to study the renoprotective potential
of P. cubeba, a 47 years old male patient diagnosed with
hypertension induced chronic kidney disease (CKD) and altered
serum creatinine level which was unable to revert to normal
levels using the conventional medication, was orally given
two capsules of P. cubeba at 4 g/day for 6 weeks (90). This
resulted in a significant improvement in subjective symptoms
(anorexia and fatigue) as well as the objective parameters of
the disorder (blood urea, serum creatinine and urine routine
and microscopy). In addition, no adverse effects were observed
during and after the study. It was concluded that P. cubeba
boost the effectiveness in reducing serum creatinine level
and in increasing estimated glomerular filtration rate (eGFR)
and may help reduce further complications related to renal
parenchymal damage.
In another study, the antilithiatic activity of the
hydroalcoholic extract of P. cubeba fruits was investigated
in male Sprague Dawley rats (91). Animals having received
the extract showed a significant decrease in crystals level in
urine. Moreover, a reduction in serum creatinine and urea was
also observed. Interestingly, magnesium in animals’ urine was
increased while sodium, calcium, phosphorus, and chloride
were significantly decreased. Likewise, histopathological
examination showed a clear improvement in kidney tissue
in treated rats with P. cubeba extract following induction of
urolithiasis by ethylene glycol and ammonium chloride. This
study strongly suggests that P. cubeba could be of significant
utility in inhibiting calcium oxalate urolithiasis. Comparatively,
streptozotocin induced diabetic Wister albino rats treated by
the root aqueous extract of P. longum maintained the normal
activities of hepatic [serum glutamic oxaloacetic transaminase
(SGOT), serum glutamic pyruvic transaminase (SGPT),
alkaline phosphatase (ALP)] and renal (serum creatinine and
urea) functional markers. This showcases the protective and
biosafety roles of P. longum extract against diabetes induced
liver and kidney damages. In addition, the extract elicited
an antihyperlipidemic activity demonstrated by a significant
decrease in the total cholesterol (TC), very low density
lipoprotein (VLDL), triglycerides (TG), low density lipoprotein
(LDL), and an increase in the high density lipoprotein (HDL)
(92). Besides, P. cubeba essential oil was also investigated for
antihyperuricemic activity and showed strong effect against
xanthine oxidase (IC50 = 54.87 µg/mL) compared to P. nigrum
EO (IC50 = 77.11 µg/mL) (7).
Melanogenesis activity
The hydroethanolic extract of P. cubeba fruits was evaluated
for melanogenesis stimulation activity using cultured murine
B16 melanoma cells. At 10 mg/ml, the extract enhanced both
intracellular and extracellular melanin contents comparatively
to the negative control. In contrast, no significant effect was
observed on cell proliferation rate. This stimulatory effect on
melanin was attributable to the presence of cubebin, a known
constituent of P. cubeba fruits (93). Comparatively, the extract of
P. methysticum and P. nigrum also showed a strong stimulatory
activity on melanogenesis. Following up these findings, guided
bioassay allowed the isolation of two kavalactones yangonin
and 7,8-epoxyyangonin from P. methysticum. When tested, both
kavalactones significantly stimulated the melanogenesis in B16
melanoma cells (93). Nevertheless, more in deep studies are
recommended to uncover the molecular targets and underpin
the mechanisms of action.
Antidepressant activity
The antidepressant potential of P. cubeba EO was
investigated in vivo using Albino mice and Fluoxetine, a
selective serotonin reuptake inhibitor, as antidepressant
standard drug (94). Using forced swimming method, animals
treated with essential oil gained weight, exhibited more
mobility, and showed less immobility comparedatively to
the mice treated with Fluoxetine. This reduction in passive
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behavior in animals highlighted the antidepressant-like
effect of P. cubeba essential oil. Interestingly, piperine
from P. nigrum was studied for its antidepressant-like
effect using corticosterone-induced model of depression
in mice for 3 weeks. Relative to the control animals, those
treated with piperine showed a significant decrease in
sucrose utilization and an increase in immobility time.
In addition, it maintained the levels of brain-derived
neurotrophic factor protein and mRNA (95). This demonstrates
the antidepressant-like effect of piperine. In another
recent study, seven compounds often found in P. nigrum
(paprazine, pellitorine, piperine, sylvamide, cepharadione
A, piperolactam D, and 10-tricosanone) were docked
against two receptors namely the potassium channel and
human serotonin transporter to assess their action on the
anxiolytic and antidepressant activities observed in vivo.
Results showed that tested compounds interact with these
target proteins with docking scores ranging from 1.0 to 7.9
kcal/mol indicating that they are likely responsible for the
antidepressant activity (96). Nevertheless, as the antidepressant
activity is still poorly explored, further experiments with
different Piper species, compounds, methods, and in vivo
models are needed.
Insecticidal activity
P. cubeba, especially its EO, was evaluated for its plant-based
insecticides activity (97). It was proved that at 0.003125%, the
EO significantly repelled Sitophilus oryzae adults. This effect
was more potent than those induced by pure compounds α-
pinene and β-caryophyllene. Following fumigation, the EO
was the most potent in causing lethality of S. oryzae adults
(LC50 = 1.07 mL cm3air). Comparatively, the EO and
α-pinene exhibited more toxic effect compared to Zingiber
officinale EO and β-caryophyllene. The noticed insecticidal
activity was attributed to the ability of the EO to inhibit
acetylcholinesterase enzyme (AchE) in fumigated rice weevil
(S. oryzae). Moreover, the oviposition of Callosobruchus sp.
was significantly reduced after fumigation with P. cubeba
EO. Similarly, a combination of 4-methyl-3-heptanol and
P. cubeba EO was revealed to be more effective as a bait
for Scolytus scolytus than multilure traps. This was due
to the synergetic action between α-cubebene and 4-methyl-
3-heptanol (98). Many other Scolytinae species including
Xyleborini and Corthylini tribes were presented to be sensitive
to P. cubeba based compounds such as α-copaene, α-
cubebene, α-humulene, and calamenene. Similarly, myristicin
(4-methoxy-6[2-propenyl]-1,3-benzodioxole) isolated from the
hexane fraction of P. mullesua D. Don fruits induced significant
toxicity against the 4th instar larvae of Spilarctia obliqua after
24 h of topical application (LD 50 = 104 µg/larva) (99).
Additionally, it was showed that in Piper genus, piperamides
are the major compounds with the strongest insecticidal
activity. Many extracts from P. nigrum,P. guineense, and
P. tuberculatum were shown to be active against insect pests
(100). In conclusion, Piper plants and compounds constitute
an innovative source of biopesticide agents for controlling
insects out-breaks.
Moreover, P. cubeba largely inhibited the germination and
growth of tow weeds (Bidens pilosa and Echinochloa crus-
galli). Noteworthy, P. cubeba EO reduced photosynthesis in the
two weeds while lipid peroxidation electrolyte leakages were
increased at 1.93 mg/mL (7).
Biological activities of isolated
compounds
Nature is the storehouse of many active compounds
that we are currently using as pharmaceuticals. P. cubeba
synthesizes many secondary metabolites, among them
hinokinin, cubebin and cubebin derivatives that are reported
to be the most pharmacologically active compounds. These
compounds exhibited many biological activities mainly
antimicrobial, anticancer, antimutagenic, antiparasitic, ovicidal,
and anticholinesterase (Table 7). In fact, lignans from P. cubeba
were shown to alter the expression of PTGS2 and MMP2
proteins in head and neck cancer cells (12). Additionally,
()-cubebin derivatives, ()-hinokinin, and ()-O-benzyl
cubebin (OBZ) at 40 mg/kg inhibited the inflammation in vivo
induced by injection of either PGE2 or dextran into the paw
of animals in comparison to indomethacin, the reference
standard (101). Besides these activities, ()-cubebin was found
to exert a vasorelaxant effect mediated by the NO/cGMP
signaling pathway without prostacyclin participation (102). In
another study, sixteen compounds were isolated from P. cubeba
extracts based on their antioxidant potential to scavenge free
radicals, hydroxyl radical, superoxide anion radical, and DPPH
(103). It was mainly found that crotepoxide was the most
active against 5,5-dimethyl-1-pyrroline-N-oxide-OH with up
to 57% inhibition. In contrast, less inhibitory activity was
noticed using other compounds such as 10-acetoxychavicol
acetate, deoxypipoxide and 3-(30,20,50-trimethoxyphenyl)
pyrrolidine. Moreover, several compounds including 5,6-
dehydrokawain, benzyl benzoate, 10-acetoxychavicol acetate,
deoxypipoxide, and 5,7,30,40-tetrametoxyflavone exhibited
superoxide dismutase (SOD)-like activity through their ability
to deliver protons (103).
General discussion
The present review comprehensively summarized
the available literature on the uses of P. cubeba and its
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TABLE 7 Biological activities of the isolated compounds from P. cubeba.
Compound name Bioactivity Results References
()-Cubebin MIC assay against E. MIC = 0.20–0.35 mM (118)
()-Hinokinin faecalis, S. salivarius, S. sanguinis, S. mitis,
S. mutans, S. sobrinus, C. albicans
MIC = 0.25–0.32 mM
()-O-(N,N-dimethylaminoethyl)-cubebin MIC = 0.19–0.31 mM
()-O-Benzyl cubebin MIC = 0.18–0.31 mM
()-6,6’-Dinitrohinokinin MIC = 0.18–0.30 mM
5-methoxy-yatein Antiparasitic activity against Schistosoma
mansoni worms
At 10–100 µM, 100% of parasites
presented a motor activity decrease
(119)
()-Hinoquinin; ()-cubebin;<