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Black Pepper (
Piper nigrum
) and Its
Bioactive Compound, Piperine
Krishnapura Srinivasan
Black pepper (Piper nigrum), an Indian native spice, has been widely
used in human diet for several thousands of years. It is valued for its
characteristic sharp and stinging qualities attributed to the alkaloid
piperine. While it is used primarily as a food adjunct, black pepper is
also used as a food preservative and as an essential component in tradi-
tional medicines in India and China. Since the discovery of black
pepper’s active ingredient, piperine, the use of black pepper has caught
the interest of modern medical researchers. Many physiological effects
of black pepper, its extracts or its bioactive compound, piperine, have
been reported in recent decades. By stimulating the digestive enzymes
of the pancreas, piperine enhances digestive capacity and significantly
reduces gastrointestinal food transit time. Piperine has been documented
to enhance the bioavailability of a number of therapeutic drugs as well
as phytochemicals through its inhibitory influence on enzymatic drug
biotransforming reactions in liver and intestine. It strongly inhibits
hepatic and intestinal aryl hydrocarbon hydroxylase glucuronyl trans-
ferase. Most of the clinical studies on piperine have focused on its effect
on drug metabolism. Piperine’s bioavailability enhancing property is
also partly attributed to increased absorption as a result of its effect on
the ultrastructure of the intestinal brush border. Piperine has been
demonstrated in in vitro studies to protect against oxidative damage by
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inhibiting or quenching reactive oxygen species. Black pepper or piper-
ine treatment has also been evidenced to lower lipid peroxidation in vivo
and beneficially influence antioxidant status in a number of experimen-
tal situations of oxidative stress. Piperine has also been found to possess
anti-mutagenic and anti-tumor influences. Clinical studies are limited,
but several have reported the beneficial therapeutic effects of black pep-
per in the treatment of smoking cessation and dysphagia.
INTRODUCTION
Black pepper (Piper nigrum) is one of the most widely used among
spices, valued for its characteristic sharp and stinging qualities. It belongs
to the family Piperaceae, cultivated for its fruit (berries) that are usually
dried and used as a spice and seasoning. Black pepper is native to
Southern India and is extensively cultivated in this tropical region. The
word “pepper” is derived from the Sanskrit “Pippali”, meaning long pep-
per. Black pepper (“Maricha” in Sanskrit) is known by other names in the
local dialect as “Milagu” (Tamil), “Kuru Mulagu” (Malayalam),
“Miriyam” (Telugu), “Miriya Konu” (Konkani), and “Kari Menasu”
(Kannada). The fruit, also known as peppercorn, is dark red when fully
mature, and a small black wrinkled drupe 5 mm in diameter when dried.
Black pepper is produced from the green unripe berries of the pepper plant
by briefly cooking in hot water. The heat ruptures cell walls in the fruit,
activating the browning enzymes during drying. Cooked berries are dried
in the sun for several days, during which the fruit around the seed shrinks
and darkens into a thin, wrinkled black layer.1
White pepper, which is commonly found in Western countries, also
comes from the same plant: it consists of the seed only with the outer fruit
removed. This is usually accomplished by soaking fully ripe pepper
berries in water for about a week, during which the flesh of the fruit soft-
ens and decomposes; rubbing off the skin results in the naked seed, which
is then dried. Ground black pepper, usually referred to simply as “pepper”,
may be commonly found on nearly every dinner table, often alongside
table salt, in some parts of the world. Dried ground pepper is one of the
most common spices in European cuisine and its descendants in other
parts of the world.1
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TRADITIONAL USES
Black pepper has been used as a spice in India since prehistoric times: it
has been known to Indian cooking since at least 2000 BC.2Peppercorns
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Black Pepper and Its Bioactive Compound, Piperine 3
Table 1. Influence of black pepper and piperine on the gastrointestinal system.
System Remarks Reference
Digestive stimulant action
Rats a) Stimulation of digestive enzymes of pancreas by dietary piperine 14
b) Stimulation of digestive enzymes of intestine by dietary piperine 15
c) Oral administration of piperine increases biliary bile acid secretion 13
Influence on intestinal motility and food transit time
Humans a) Increased orocecal transit time after black pepper consumption 24
Rats a) Gastrointestinal food transit time shortened by dietary piperine 27
b) Piperine inhibits gastric emptying of solids/liquids 25
Mice a) Piperine inhibits gastrointestinal transit 25
b) Piperine dose-dependently delays gastrointestinal motility 26
Effect on gastric mucosa
Humans a) Black pepper causes increase gastric parietal and pepsin 16
secretion and increased gastric cell exfoliation in humans
Rats a) Black pepper increases gastric acid secretion in anesthetized rats 18
b) Piperine increases gastric acid secretion 19
c) Piperine has protective action against stress-induced gastric ulcer 17
Mice a) Piperine has protective action against stress-induced gastric ulcer 17
Antidiarrheal property
Mice a) Piperine inhibits diarrhea produced by castor oil, arachidonic 20
acid and magnesium sulfate
b) Piperine reduces castor oil-induced intestinal fluid accumulation 21
in intestine
Influence on absorptive function
Rats a) Piperine stimulates γ-glutamyl transpeptidase activity and 22
enhanced uptake of amino acids in isolated epithelial cells
of rat jejunum
b) Piperine modulates membrane dynamics and permeation 23
characteristics, increasing absorptive surface and induction of
synthesis of proteins associated with cytoskeletal function
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were a much prized trade good, often referred to as “black gold.” Black
pepper, along with other spices from India and the Far East, changed the
course of world history. Black pepper has been known in China since the
2nd century BC. The pepper trade was first dominated by China, who
imported black pepper in mass quantities during the 14th to 16th cen-
turies. Pepper was introduced into Sumatra at the beginning of the 15th
century, where pepper cultivation and mass production grew exponen-
tially. It is also recorded that the preciousness of these spices led to
European efforts to find a sea route to India and consequently to the
European colonial occupation of that country, as well as to the European
discovery and colonization of the Americas. Black pepper is referred to as
“King of Spices” and represents one of India’s major commodities.3
It is also known that black pepper was once used as a food preserva-
tive. Although it is difficult to believe that in the Middle Ages pepper was
used as a preservative for meat, sure enough piperine, the compound that
gives pepper its spiciness, has some antimicrobial properties, but not at
the concentrations present when pepper is used as a spice. However, pep-
per and other spices probably did play a role in improving the taste of
long-preserved meats. Moreover, in the Middle Ages, pepper was a luxury
item, affordable only to the wealthy. Having been an item exclusively for
the rich, pepper started to become more of an everyday seasoning among
those of more average means.
PEPPER AS ANCIENT MEDICINE
Black pepper is historically used not only in human diets but also in tra-
ditional medicines and home remedies.4The use of black pepper in
medicine in India dates back thousands of years. Long pepper (Piper
longum), being stronger, was often the preferred medication although both
were used. Black pepper figures in remedies in Ayurveda, Siddha and
Unani medicine in India. The 5th century Syriac Book of Medicines pre-
scribes black pepper (and long pepper) for such illnesses as constipation,
diarrhea, earache, gangrene, heart disease, hernia, hoarseness, indigestion,
insect bites, insomnia, joint pain, liver problems, lung disease, oral
abscesses, sunburn, tooth decay, and toothaches. Black pepper was relied
upon to treat specific conditions such as diarrhea and fevers, but it appears
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that its extensive generalized use was to enhance the effects of many
herbal remedies.4
Ayurvedic physicians have been prescribing long pepper and black pep-
per (both of which are now known to contain piperine) for thousands of
years, a practice which may have enhanced the pharmacological actions of
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Black Pepper and Its Bioactive Compound, Piperine 5
Table 2. Influence of piperine on the drug metabolizing enzyme system.
System Remarks Reference
In vitro a) Inhibition of aryl hydroxylation, N-demethylation, 28
O-deethylation and glucuronidation in vitro by piperine
b) Decreased UDP-glucuronic acid concentration and rate of 29
glucuronidation in isolated epithelial cells of guinea pig
small intestine by piperine
c) Inhibition of aryl hydroxylase and O-deethylase activities 32
by piperine in vitro in pulmonary microsomes
d) Suppression of aryl hydroxylation in cell culture is 34
mediated by direct inter-action of piperine with
cytochrome P450 and not by downregulation of its gene
expression
e) Piperine decreases the activities of liver microsomal aryl 36
hydroxylase, N-demethylase and UDP-glucuronosyl
transferase and cytochrome P450
Rats a) Lower aryl hydroxylase and UDP-glucuronyl transferase 28
activities, prolonged hexobarbital sleeping time in
piperine treated rats
b) Inhibition of aryl hydroxylase and O-deethylase activities 32
by piperine in vivo in pulmonary microsomes
c) Decreased activities of hepatic microsomal cytochrome 30
P450, N-demethylase, aryl hydroxylase by intragastric/
intraperitoneal piperine
d) Inhibition of UDP-glucose dehydrogenase and UDP- 33
glucuronyl transferase in liver and intestine by piperine
e) Lowered activity of N-demethylase, UDP-glucuronosyl 36
transferase and NADPH-cytochrome-C reductase as a
result of piperine feeding
Guinea a) Inhibition of UDP-glucose dehydrogenase and UDP- 33
pigs glucuronyl transferase in liver and intestine by piperine
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other compounds in traditional herbal medicines. It is a vital ingredient of
many remedies in the traditional Ayurvedic system of medicine in India.
Black pepper is a component of “Trikatu” (three acrids) along with long pep-
per (Piper longum) and ginger (Zingiber officinale) in equal proportions.
Trikatu is widely used in combination with other Ayurvedic medications
according to the ancient Ayurvedic Materia Medica (600–300 BC). Very few
compound prescriptions are free from these three acrids.5Trikatu aims to cor-
rect the imbalance of the three “doshas” (psychophysical components of the
human body) that can lead to disease.6Black pepper is specifically cited in
Ayurveda to internally treat fevers, gastric and abdominal disorders, and uri-
nary problems.6Medicinal external treatments with black pepper include
treatments for rheumatism, neuralgia, and boils.6Piper nigrum is also used
to treat alopecia.5Possible uses of black pepper in Indian folk medicine
include the treatment of respiratory diseases, dysentery, pyrexia, and insom-
nia.6Black pepper is part of an herbal folk remedy relied upon by mothers to
treat their children’s diarrhea.7This wide-ranging use of black pepper in
India is unprecedented in other medical systems and areas of the world.
Once black pepper reached China, it was incorporated into traditional
Chinese medicine. Pepper is cited for its digestive stimulant action — to
make food enter the large intestine channels to “warm the middle, disperse
cold, drive the food downward while dispelling phlegm, wind-cold, and
relieving diarrhea.”8This is caused by stomach cold, characterized by vom-
iting, diarrhea, and abdominal pain. Black pepper has been used in China
as a folk remedy for epilepsy. A popular Chinese folk remedy for epilepsy
calls for a dried powder consisting of one radish and 99 peppercorns.9
Black pepper is also used as contraceptive in Assam (A north-eastern state
in India) folk medicine.
CHEMICAL CONSTITUENTS
The spiciness of black pepper, which is characterized by its distinct sharp
and stinging qualities, is due to the alkaloid compound piperine, found
both in the outer fruit and in the seed.1Refined piperine is about 1% as hot
as the capsaicin of red chili pepper. The outer fruit layer left on black pep-
per also contains important odor-contributing terpenes including pinene,
sabinene, limonene, caryophyllene, and linalool, which give citrusy,
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woody, and floral notes. Pepper contains small amounts of safrole, a
mildly carcinogenic compound. The bioactive and pungent ingredient of
black pepper was identified as piperine and isolated in 1820 by the Dutch
chemist Hans Christian Orstedt.1
Many beneficial physiological effects of black pepper, its extracts or
its major active principle, piperine, have been reported in recent decades
and have been reviewed.10
DIVERSE EXPERIMENTALLY VALIDATED
BENEFICIAL PHYSIOLOGICAL EFFECTS
OF BLACK PEPPER AND PIPERINE
Influence on the Gastrointestinal System
Digestive stimulant action
It is a general perception that aromatic and pungent spices, by imparting
flavor and appealing taste to foodstuffs, enhance salivary and gastric
secretions. Glatzel, studying the effect of spices on the secretion and
composition of saliva in human subjects, observed that black pepper and
other spices enhance the secretion of saliva and the activity of salivary
amylase.11 The digestive stimulant action of spices is exerted through:
(i) a beneficial stimulation of the liver to produce and secrete bile rich
in bile acids, which play a very important role in fat digestion and
absorption, or (ii) a beneficial stimulation of the activities of enzymes of
pancreas and intestine that participate in digestion.12 Black pepper and
its active principle, piperine, examined for their effect on bile secretion
as a result of both a continued intake through the diet for a period of
time and as a one-time exposure orally in experimental rats, did not
show any beneficial stimulatory influence on bile acid production by the
liver and its secretion into bile.13 On the other hand, oral administration
of piperine as a single dose significantly increased bile acid secretion.
The influence of dietary intake of piperine (at levels corresponding to
about five times the average dietary intake of black pepper by the Indian
population) on the pancreatic digestive enzymes and the terminal diges-
tive enzymes of the small intestinal mucosa have been examined in
experimental rats.14,15 Significantly increased activities of pancreatic
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lipase, amylase, chymotrypsin and trypsin were observed as a result of
dietary intake of piperine in these experimental rats.14 Such beneficial
influence of this spice on the activity of these enzymes was not evident
when administered as a single oral dose. Piperine also prominently
enhanced the activity of intestinal lipase and amylase in animals given
single oral doses of piperine.15
Effect on gastric mucosa
Clinical studies: Pungent spices have long been implicated as a cause of
gastric mucosal injury; their long-term effect on the gastric mucosa is
still less known. In a single-dose study, the effects of black pepper on
the gastric mucosa were assessed using double-blind intragastric admin-
istration of the spice (1.5 g) to healthy human volunteers, with aspirin
(655 mg) as positive control.16 Black pepper caused significant increases
in parietal secretion, pepsin secretion, and potassium loss. Gastric cell
exfoliation (as reflected in DNA loss in gastric contents) was increased
after black pepper administration and mucosal microbleeding was also
seen. These effects of black pepper on gastric mucosa were similar to
aspirin.
Animal studies: On the other hand, the protective action of piperine
against experimental gastric ulcer has been evidenced in rats and mice
wherein the gastric mucosa damage was induced by stress, indometacin,
HCl, and pyloric ligation.17 Piperine at 25, 50, 100 mg/kg i.g. protected
animals from gastric ulceration in a dose-dependent manner. Piperine
inhibited the volume of gastric juice, gastric acidity, and pepsin activity.
Black pepper has been reported to significantly increase gastric acid
secretion in anesthetized rats.18 Piperine has been shown to produce a
dose-dependent (20–142 mg/kg) increase in gastric acid secretion in albino
rats.19 Involvement of cholinergic receptors in the observed piperine-
induced increase in gastric acid secretion has been ruled out as the effect
of piperine was significantly antagonized by cimetidine (1 mg/kg) but not
by atropine (1 mg/kg). There is, however, an indication that increased
acidity induced by piperine could be due to stimulation of histamine H2
receptors by this spice compound.
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Antidiarrheal property
Animal studies: Peppers are added in traditional antidiarrheal formula-
tions of different herbs. In a study undertaken in experimental mice, the
antidiarrheal activity of piperine against diarrhea produced by castor oil,
MgSO4and arachidonic acid has been evidenced at 8 and 32 mg/kg p.o.
doses.20 Piperine’s inhibition of castor oil-induced enteropooling suggests
its inhibitory effect on prostaglandins. Piperine (2.5–20 mg/kg i.p.) dose-
dependently reduced castor oil-induced intestinal fluid accumulation
in experimental mice.21 It was further understood that piperine reduces
castor oil-induced fluid secretion with a mechanism involving capsaicin-
sensitive neurons but not capsazepine-sensitive vanilloid receptors.
Influence on absorptive function
Animal studies: The effect of piperine on the absorptive function of the
intestine has been studied in in vitro experiments, showing that piperine
(25–100 µM) significantly stimulated γ-glutamyl transpeptidase (γ-GT)
activity and enhanced the uptake of amino acids in freshly isolated epithe-
lial cells of rat jejunum.22 The kinetic behavior of γ-GT towards substrate
and acceptor was altered in the presence of piperine, suggesting that
piperine may interact with the lipid environment to produce effects lead-
ing to increased permeability of the intestinal cells. It is hypothesized that
piperine’s bioavailability-enhancing property may be partly attributed to
increased absorption.23 Piperine also caused an increase in intestinal brush
border membrane fluidity and stimulated leucine amino peptidase and
glycyl-glycine dipeptidase activity due to the alteration in enzyme kinet-
ics. This suggests that piperine could modulate the membrane dynamics
due to its apolar nature by interacting with surrounding lipids and
hydrophobic portions in the protein vicinity, which may decrease the ten-
dency of membrane lipids to act as stearic constraints to enzyme proteins
and thus modify enzyme conformation. Ultrastructural studies with piper-
ine showed an increase in microvilli length with a prominent increase in
free ribosomes and ribosomes on the endoplasmic reticulum in entero-
cytes, suggesting that synthesis or turnover of cytoskeletal components
or membrane proteins may be involved in the observed effect. Thus,
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piperine may induce alterations in membrane dynamics and permeation
characteristics, along with induction of the synthesis of proteins associ-
ated with cytoskeletal function, resulting in an increase in the absorptive
surface, thus assisting efficient permeation through the epithelial barrier.
Influence on gastrointestinal motility and food transit time
Clinical studies: In a study of the effect on small intestinal peristalsis
evaluated by measuring orocecal transit time utilizing the lactulose hydro-
gen breath test in healthy subjects, an increase in orocecal transit time was
observed after black pepper (1.5 g) consumption.24
Animal studies: Piperine has been found to inhibit gastric emptying (GE)
of solids/liquids in rats and gastro-intestinal transit (GT) in mice in a
dose- and time-dependent manner.25 It significantly inhibited GE of
solids and GT at the doses extrapolated from humans (1 mg/kg and
1.3 mg/kg p.o. in rats and mice, respectively). One week oral treatment
of 1 mg/kg and 1.3 mg/kg in rats and mice, respectively, did not produce
a significant change in activity as compared to single-dose administra-
tion. The GE inhibitory activity of piperine is independent of gastric acid
and pepsin secretion. Piperine, which activates vanilloid receptors
(0.5–20 mg/kg i.p.) dose-dependently, delayed gastrointestinal motility
in mice.26 The inhibitory effect of piperine (10 mg/kg) was strongly atten-
uated in capsaicin-treated (75 mg/kg in total, s.c.) mice. The study
indicated that the vanilloid ligand piperine can reduce upper gastroin-
testinal motility. The effect of piperine involves capsaicin-sensitive
neurones but not vanilloid receptors.
The gastrointestinal food transit time in experimental rats has been
shown to be significantly shortened by dietary piperine.27 The reduction in
food transit time produced by dietary piperine roughly correlated with its
beneficial influence either on digestive enzymes or on bile secretion.12
Thus, dietary piperine, which enhanced the activity of digestive enzymes,
also markedly reduced the food transit time at the same level of con-
sumption. This reduction in food transit time could probably be attributed
to acceleration in the overall digestive process as a result of increased
availability of digestive enzymes.
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Inhibitory Influence of Piperine on Drug Metabolizing
Enzyme System
In the context of piperine having been reported to enhance drug bioavail-
ability, Atal et al. studied the interaction of piperine with drug
biotransforming reactions in hepatic tissue in vitro and in vivo.28 Piperine
inhibited hydroxylation of aryl hydrocarbon, N-demethylation of ethyl-
morphine, O-deethylation of 7-ethoxycoumarin and glucuronidation of
3-hydroxybenzo (α) pyrene (3-OH-BP) in rat liver in vitro in a dose-
dependent manner. Piperine caused noncompetitive inhibition of hepatic
microsomal aryl hydrocarbon hydroxylase (AHH) from untreated and
3-methylcholanthrene-treated rats with a Kiof 30 µM. Similarly, the kinet-
ics of inhibition of ethylmorphine-N-demethylase from control rat liver
exhibited noncompetitive inhibition with a Kmof 0.8 mM and Kiof 35 µM.
These studies demonstrate that piperine is a nonspecific inhibitor of drug
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Black Pepper and Its Bioactive Compound, Piperine 11
(a) Black pepper (b) Pepper plant
O
O
H2C
N
O
(c) Chemical structure of Piperine
Fig. 1. Photographs of (a) the spice, and (b) the spice plant. (c) the chemical structure of
piperine.
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metabolism which shows little discrimination between different
cytochrome P450 forms. Oral administration of piperine in rats strongly
inhibited the hepatic AHH and UDP-glucuronyl transferase activities, the
inhibition of AHH being observed within 1 hr and restored to normal by
6 hrs. Pretreatment with piperine prolonged hexobarbital sleeping time
and zoxazolamine paralysis time in mice. These results demonstrate that
piperine is a potent inhibitor of drug metabolism. The basis of inhibition
of glucuronidation by piperine has been explored by examining the rate of
glucuronidation of 3-OH-BP and UDP-glucuronic acid (UDPGA) content
in the intact isolated epithelial cells of the guinea-pig small intestine.29 It
was found that glucuronidation of 3-OH-BP was dependent on duration of
incubation, cellular protein and endogenous UDPGA concentration.
Piperine also caused a concentration-related decrease in UDPGA content
and the rate of glucuronidation in these cells. Piperine also caused non-
competitive inhibition of hepatic microsomal UDP-glucuronyltransferase
with a Kiof 70 µM. The study demonstrated that piperine modifies the rate
of glucuronidation by lowering the endogeneous UDPGA content and
also by inhibiting the transferase activity.
Although an increase in hepatic microsomal cytochrome P450 and
cytochrome b5, NADPH-cytochrome creductase, benzphetamine
N-demethylase, aminopyrine N-demethylase and aniline hydroxylase
was observed 24 hrs following intra-gastric administration of piperine
(100 mg/kg) in adult Sprague–Dawley rats, a higher intra-gastric dose
(800 mg/kg) or i.p. (100 mg/kg) dose of piperine produced a significant
decrease in the levels of cytochrome P450, benzphetamine N-demethylase,
aminopyrine N-demethylase and aniline hydroxylase 24 hrs after treat-
ment.30 An i.p. administration of rats with piperine (100 mg/kg) produced
a significant decrease in hepatic cytochrome P450 and activities of ben-
zphetamine N-demethylase, aminopyrine N-demethylase and aniline
hydroxylase 1 hr after the treatment.31 Twenty-four hours later, these
parameters along with cytochrome b5and NADPH-cytochrome creduc-
tase remained depressed in piperine-treated rats. This suggested that the
effect of piperine on hepatic mixed-function oxidases is monophasic.
Piperine caused concentration-related non-competitive inhibition
in vitro (50% at 100 µM) of AHH and 7-ethoxycoumarin deethylase activ-
ities in lung microsomes of rats and guinea pigs.32 In vivo, piperine given
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at a dose of 25 mg/kg body weight to rats caused a maximal inhibition at
1 hr of both the enzymes, while only AHH returned to the normal value
within 4 hrs. Similarly, upon daily treatment of piperine (15 mg/kg body
weight) to rats for 7 days, deethylase activity was consistently inhibited,
while AHH showed faster recovery. Piperine thus appeared to cause dif-
ferential inhibition of two forms of cytochrome P450 and thus would
accordingly affect the steady-state level of those drugs metabolized by
these pulmonary forms of cytochrome P450.
Piperine caused a concentration-related strong non-competitive
inhibition of UDP-glucose dehydrogenase (UDP-GDH) (50% at 10 µM)
reversibly and equipotently in rat and guinea pig liver and intestine.33
However, the UDPGA contents were decreased less effectively by
piperine in isolated rat hepatocytes compared with enterocytes of
guinea pig small intestine. Piperine at 50 µM caused a marginal
decrease of UDPGA in hepatocytes when the rate of glucuronidation of
3-OH-BP decreased by about 40%. Piperine did not affect the rate
of glucuronidation of 4-OH-biphenyl in rat liver, whereas that of
3-OH-BP was impaired significantly. In guinea pig small intestine,
both these activities were inhibited significantly, requiring less than
25 µM piperine to produce a more than 50% inhibition of UDP-
glucuronyl transferase. The results suggest that piperine is a potent
inhibitor of UDP-GDH and it exerts stronger effects on intestinal
glucuronidation than in rat liver.
By studying the modulation of B(α)p metabolism and regulation of
cytochrome CYP1A1 gene expression by piperine in 5L cells in culture,
it has been observed that piperine mediated inhibition of AHH activity,
and that the consequent suppression of the procarcinogen activation is the
result of direct interaction of piperine with cytochrome P4501A1-protein
and not because of down regulation of its gene expression.34 Piperine was
evaluated for beneficial effects in Alzheimer’s disease by studying the
potential for herb-drug interactions involving cytochrome P450, UDP-
glucuronosyl transferase, and sulfotransferase enzymes. Piperine was a
relatively selective noncompetitive inhibitor of CYP3A (IC50 of 5.5 µM,
Kiof 5.4 µM) with less effect on other enzymes evaluated (IC50 >29 µM).35
Piperine inhibited recombinant CYP3A4 much more potently (more than
five fold) than CYP3A5.
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The effect of dietary supplementation of piperine (0.02%) on the
activities of the liver drug-metabolizing enzyme system has been exam-
ined in rats.36 Piperine significantly stimulated the activity of aryl
hydroxylase. The activity of N-demethylase, UDP-glucuronosyl trans-
ferase and NADPH-cytochrome creductase activity was significantly
lowered as a result of piperine feeding, while the levels of hepatic micro-
somal cytochrome P450 and cytochrome b5were not influenced by
piperine. Piperine also significantly decreased the activities of liver
microsomal AHH, N-demethylase and UDP-glucuronosyl transferase in
vitro at a 1 ×10−6mol/L level in the assay medium. Piperine also brought
about a significant decrease in liver microsomal cytochrome P450 when
included at 1 ×10−6mol/L.
The modifying potential of black pepper on the hepatic biotransfor-
mation system has been assessed in mice fed on a diet containing 0.5%,
1% and 2% black pepper for 10 and 20 days.37 Data revealed a significant
and dose-dependent increase in glutathione S-transferase and sulfhydryl
content in the experimental groups on the 1% and 2% black pepper diets.
Elevated levels of cytochrome b5and cytochrome P450 were also signif-
icant and dose dependent. As a potential inducer of the detoxication
system, the possible chemopreventive role of black pepper in chemical
carcinogenesis was suggested.
Piperine Enhances the Bioavailability of Drugs and
Phytochemicals
Clinical studies: Piperine, the alkaloid constituent of both black and long
pepper, is now established as a bioavailability enhancer of various struc-
turally and therapeutically diverse drugs and other substances. The
potential of piperine to increase the bioavailability of drugs in humans is
of great clinical significance. Most of the clinical trials done on black
pepper have shown that piperine increases levels of certain medications:
phenytoin (an epileptic treatment), propranolol (used for hypertension and
stage fright), rifampicin (a tuberculosis medication), theophylline (lung
medication), and even coenzyme Q10. This observed effect is due to
the inhibitory interaction of piperine with cytochrome P450 enzymes
of the liver and gastrointestinal tract that are also involved in drug
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metabolism: CYP1A2, CYP1A1, CYP2D6, CYP3A4; P-glycoprotein
(P-gp) is also affected.38 Since piperine inhibits both P-glycoprotein and
CYP3A4 expressed in enterocytes and hepatocytes, it contributes to a
major extent to first-pass elimination of many drugs.38
The scientific basis of the use of the trikatu group of acrids (long pep-
per, black pepper and ginger) in a large number of prescriptions in the
indigenous Ayurvedic system of medicine in India has been evaluated by
Atal et al.39 The observed increase of over 200% in the blood levels of the
test drug vasicine by Piper longum and of the blood levels of the test drug
sparteine by over 100% under the influence of piperine in a clinical study
suggested that these acrids have the capacity to increase the bioavailabil-
ity of certain drugs. The authors concluded that the trikatu group of drugs
increases bioavailability of drugs either by promoting rapid absorption
from the gastrointestinal tract or by protecting the drug from being metab-
olized in its first passage through the liver after being absorbed, or by a
combination of these two mechanisms. The effect of piperine on the
bioavailability and pharmacokinetics of propranolol and theophylline has
been examined in a crossover study wherein subjects received a single
oral dose of propranolol (40 mg) or theophylline (150 mg) alone or in
combination with piperine (20 mg/day for 7 days).40 An enhanced sys-
temic availability of oral propranolol and theophylline was evidenced as
a result of piperine treatment.
A pharmacokinetic study has examined the effect of piperine, a
known inhibitor of hepatic and intestinal glucuronidation on the bioavail-
ability of curcumin, the bioactive ingredient of the spice? turmeric
administered with piperine in healthy human volunteers.41 The human
study was done in a cross-over design with two weeks separating two clin-
ical testing sessions. After a dose of 2 g of curcumin taken without
piperine, serum levels were either undetectable or very low. Concomitant
administration of piperine (20 mg) produced 2000% higher concentra-
tions from 0.25 to 1 hr post-drug. The study showed that, in the dosages
used, piperine enhances the serum concentration, extent of absorption and
bioavailability of curcumin in humans. This assumes importance in the
context of the diverse medicinal properties of Curcuma longa. Black pep-
per extract consisting of 98% piperine has been evidenced to increase
plasma levels of orally supplemented coenzyme Q10 in a clinical study
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using a double-blind design.42 The relative bioavailability of 90 mg and
120 mg of coenzyme Q10 administered in a single dose or for 14 and 21
days with placebo or with 5 mg of piperine was determined by comparing
measured changes in plasma concentration. Supplementation of 120 mg
coenzyme Q10 with piperine for 21 days produced a significant, approxi-
mately 30% greater AUC than with coenzyme Q10 plus placebo.
Piperine has been reported to enhance the oral bioavailability of
phenytoin in human volunteers. The effect of a single dose of piperine in
patients with uncontrolled epilepsy on the steady-state pharmacokinetics
of phenytoin has been examined.43 Piperine (20 mg administered along
with phenytoin) increased significantly the mean plasma concentration of
phenytoin at most of the time points in patients receiving either 150 mg or
200 mg twice daily doses of phenytoin. There was a significant increase
in AUC, Cmax and Ka.
Nevirapine is a potent non-nucleoside inhibitor of HIV-1 reverse tran-
scriptase and is indicated for use in combination with other antiretroviral
agents for the treatment of HIV-1 infection. In a cross-over, placebo-
controlled study conducted in eight healthy adult males, subjects received
piperine 20 mg or placebo for 6 days, and on day 7, nevirapine 200 mg
plus piperine 20 mg or nevirapine plus placebo in a crossover fashion.44
Mean maximum plasma concentration, the area under the plasma
concentration-time curve, from 0 to 144 hrs post-dose were increased
significantly when co-administered with piperine. This evidence for
enhanced bioavailability of nevirapine when administered with piperine
suggests a possible clinical advantage arising from the bioenhancement
capabilities of piperine in the treatment of HIV infection.
Animal studies: It has been observed that intragastric cotreatment with
dietary piperine enhances the bioavailability of epigallocatechin-3-gallate
(EGCG; demonstrated to have chemopreventive activity) from green tea
in mice.45 Coadministration of 164 µmol/kg EGCG and 70 µmol/kg piper-
ine to male mice increased the plasma Cmax and area under the curve
(AUC) by 1.3-fold compared to mice treated with EGCG only. Piperine
appeared to increase EGCG bioavailability by inhibiting glucuronidation
and gastrointestinal transit. A similar effect of piperine in altering the
pharmacokinetics of phenytoin, an anti-epileptic drug, was reported from
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a study on mice.46 Pretreatment of piperine significantly delayed the elim-
ination of phenytoin. Coadministration of piperine enhanced the
bioavailability of β-lactam antibiotics, amoxycillin trihydrate and cefo-
taxime significantly in rats.47 The improved bioavailability is reflected in
various pharmacokinetic parameters, viz. tmax, Cmax, half-life and AUC, of
these antibiotics and was attributed to the effect of piperine on microso-
mal metabolizing enzymes.
When curcumin was given alone at 2 g/kg to rats, moderate serum
concentrations were achieved over a period of 4 hrs.41 Concomitant
administration of piperine (20 mg/kg) increased the serum concentration
of curcumin for a short period of 1–2 hrs post-drug. Time to maximum
was significantly increased while plasma half-life and clearance signifi-
cantly decreased, and the bioavailability was increased by 154%.
The effect of piperine on the metabolic activation and distribution of
aflatoxin B1(AFB1) in rats has been studied.48 Rats pretreated with piper-
ine accumulated considerable AFB1in plasma and in the tissues examined
as compared to the controls. Piperine had no influence on hepatic AFB1-
DNA binding in vivo, which could possibly be due to the null effect of
piperine on liver cytosolic glutathione transferase activity. Piperine-
treated rat liver microsomes demonstrated a tendency to enhance AFB1
binding to calf thymus DNA in vivo. Piperine markedly inhibited liver
microsome-catalyzed AFB1binding to calf thymus DNA in vitro, in a
dose-dependent manner.
Antioxidant Effect of Piperine
In vitro studies: Oxygen radical injury and lipid peroxidation have been
suggested as major causes of atherosclerosis, cancer and the aging
process. Piperine has been demonstrated in in vitro experiments to protect
against oxidative damage by quenching free radicals and reactive oxygen
species and inhibiting lipid peroxidation.49 Piperine is reported to have
marginal inhibitory effects on ascorbate/Fe2+-induced lipid peroxidation
in rat liver microsomes even at high concentrations (600 µM) when com-
pared to the beneficial inhibition of lipid peroxidation by antioxidants
vitamin E, t-butylhydroxytoluene and t-butylhydroxyanisole.50 Both water
and ethanol extract of black pepper exhibited strong total antioxidant
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Black Pepper and Its Bioactive Compound, Piperine 17
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activity, and significant inhibition of peroxidation of linoleic acid emul-
sion.51 Piperine is shown to be an effective antioxidant and offer
protection against oxidation of human low density lipoprotein (LDL) as
evaluated by copper ion-induced lipid peroxidation of human LDL by
measuring the formation of thiobarbituric acid reactive substance and rel-
ative electrophoretic mobility of LDL on agarose gel.52 The aqueous
extract of black pepper as well as piperine have been examined for their
effect on human PMNL 5-lipoxygenase (5-LO), the key enzyme involved
in biosynthesis of leukotrienes.53 The formation of 5-LO product 5-HETE
was significantly inhibited in a concentration-dependent manner with
IC50 values of 0.13 mg for aqueous extracts of pepper and 60 µM for
piperine. Thus, piperine from black pepper might exert an antioxidant
physiological role by modulating the 5-LO pathway.
Animal studies: Piperine treatment (10 mg/kg/day i.p. for 14 days) has
been assessed for protection against diabetes-induced oxidative stress in
streptozotocin-induced diabetic rats.54 Treatment with piperine reversed
the diabetic effects on glutathione concentration in brain, on renal glu-
tathione peroxidase and superoxide dismutase activities, and on cardiac
glutathione reductase activity and lipid peroxidation, but did not reverse
the effects of diabetes on hepatic antioxidant status. Thus, subacute treat-
ment with piperine for 14 days is only partially effective as an antioxidant
in diabetes. The ability of piperine to reduce the oxidative changes
induced by chemical carcinogens (7,12-dimethylbenzanthracene,
dimethylaminomethylazobenzene and 3-methylcholanthrene) has been
investigated in a rat intestinal model.55 A protective role of piperine
against the oxidative alterations by these carcinogens was indicated by the
observed inhibition of TBARS, a significant increase in the glutathione
levels and restoration in γ-GT and Na+, K+-ATPase activity in intestinal
mucosa. The impact of piperine on alterations of the mitochondrial
antioxidant system and lipid peroxidation in benzo(α)pyrene (B(α)p)
induced experimental lung carcinogenesis has been investigated in mice.56
Oral supplementation of piperine (50 mg/kg body weight) effectively sup-
pressed lung carcinogenesis by B(α)p as revealed by a decrease in the
extent of mitochondrial lipid peroxidation and concomitant increase in the
activities of enzymatic antioxidants and nonenzymatic antioxidant levels
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when compared to lung carcinogenesis bearing animals. This suggests
that piperine may extend its chemo-preventive effect by modulating lipid
peroxidation and augmenting the antioxidant defense system.
The effect of supplementation of black pepper (0.25 g or 0.5 g/kg
body weight) or piperine (0.02 g/kg body weight) for a period of 10 wks
on tissue lipid peroxidation, enzymic and non-enzymic antioxidants has
been examined in rats fed a high-fat diet (20% coconut oil and 2% cho-
lesterol) and it was observed that these can reduce high-fat diet-induced
oxidative stress.57 Simultaneous supplementation with black pepper or
piperine lowered TBARS and conjugated diene levels and maintained
antioxidant enzymes and glutathione levels in the liver, heart, kidney,
intestine and aorta near to those of control rats.
Antimutagenic and Tumor Inhibitory Effects
Cell line studies: Black pepper has been shown to be effective in reduc-
ing the mutational events induced by the promutagen ethyl carbamate
in Drosophila melanogaster.58 Suppression of metabolic activation or
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Black Pepper and Its Bioactive Compound, Piperine 19
MODULATION OF BIOAVAILABILITY INHIBITION OF DRUG PROTECTIVE EFFECT ON CYTO-
OF THERAPEUTIC DRUGS, PHYTO- METABOLIZING SYSTEM TOXICITY BY CARCINOGENS
CHEMICALS AND CARCINOGENS
ANTIMUTAGENIC AND ANTIOXIDANT INFLUENCE
ANTICANCER EFFECTS PIPERINE
EFFECTS ON GASTROINTESTINAL SYSTEM OTHER PHYSIOLOGICAL EFFECTS
Digestive stimulant action Decreases fertility by interference with crucial events
Influence on intestinalmotility and food transit time Anti-inflammatory activity
Effect on gastric mucosa: increased gastric secretion; Growth stimulatory effect on melanocytes
protective effect on gastric ulcer Antidepressant-like effects
Anti-diarrheal property Anticonvulsant effects
Enhances absorptive function of intestine Amelioration of dysphagia
Fig. 2. The diverse physiological effects of piperine.
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interaction with the active groups of mutagens could be mechanisms by
which this spice exerts its antimutagenic action. While studying piperine
for its immuno-modulatory and antitumor activity, piperine was found to
be cytotoxic towards Dalton’s lymphoma ascites (DLA) and Ehrlich
ascites carcinoma (EAC) cells at 250 µg/ml.59 Piperine was also found to
produce cytotoxicity towards L929 cells in culture at a concentration of
50 µg/ml. Administration of piperine (1.14 mg/animal) could inhibit solid
tumor development in mice induced with DLA cells and increase the
lifespan of mice bearing Ehrlich ascites carcinoma tumors to 59%.
The effect of piperine on the cytotoxicity and genotoxicity of afla-
toxin B1(AFB1) has been studied in rat hepatoma cells
H4IIEC3/G-(H4IIE) using cellular growth and formation of micronuclei
as endpoints.60 AFB1inhibited the growth of H4IIE cells with an ED50 of
15 nM. Piperine markedly reduced the toxicity of the mycotoxin. Piperine
reduced the AFB1-induced formation of micronuclei in a concentration-
dependent manner. The results suggest that piperine is capable of
counteracting AFB1toxicity by suppressing cytochrome P450 mediated
bioactivation of the mycotoxin. The potential of piperine to inhibit the
activity of cytochrome P450 2B1 and protect against AFB1has been
investigated in r2B1 cells (Chinese hamster cells) engineered for the
expression of rat CYP450 2B1.61 Piperine inhibited 7-methoxycoumarin
demethylase in preparations of r2B1 cells with an IC50 of 10 µM.
Piperine at 60 µM completely counteracted cytotoxicity and formation of
micronuclei by 10 µM AFB1and reduced the toxic effects of 20 µM AFB1
by more than 50%. The results suggest that (i) piperine is a potent
inhibitor of rat CYP450 2B1 activity, (ii) AFB1is activated by r2B1 cells
to cytotoxic and genotoxic metabolites, and (iii) piperine counteracts
CYP450 2B1 mediated toxicity of AFB1in the cells and might, therefore,
offer a potent chemopreventive effect against procarcinogens activated by
CYP450 2B1.
Animal studies: The antimutagenic effect of piperine has been studied par-
ticularly with respect to its influence on chromosomes in rat bone marrow
cells.62 Male Wistar rats orally administered piperine (100, 400 and 800
mg/kg body weight) were challenged with cyclophosphamide (i.p. 50 mg/kg
body weight), sacrificed 24 hrs thereafter and bone marrow samples were
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collected. Piperine at a dose of 100 mg/kg body weight gave a statistically
significant reduction in cyclophosphamide-induced chromosomal aberra-
tions, suggesting that piperine may have antimutagenic potential. Black
pepper extracts have been demonstrated to possess tumor inhibitory activ-
ity.63 The tumor reducing activity of orally administered extracts of black
pepper was studied in mice transplanted i.p. with Ehrlich ascites tumor.64
Lifespan was increased in these mice by 65%, indicating the potential use
of the spice as anti-cancer agents as well as anti-tumor promoters. The
antimetastatic activity of piperine has been demonstrated by the inhibition
of lung metastasis induced by B16F-10 melanoma cells in C57BL/6
mice.65 Simultaneous administration of the compound with tumor induc-
tion produced a significant reduction in tumor nodule formation. The
elevated levels of serum sialic acid and serum γ-GT activity in the
untreated animals were significantly reduced in the animals treated with
piperine.
The cytoprotective effect of piperine on B(α)p-induced experimental
lung cancer has been investigated in mice and it was observed that piper-
ine may extend its chemopreventive effect by modulating lipid
peroxidation and augmenting the antioxidant defense system.66 Oral
administration of piperine (100 mg/kg body weight) effectively sup-
pressed lung cancer initiated with B(?)p as revealed by the decrease in the
extent of lipid peroxidation with concomitant increase in the activities of
enzymatic antioxidants and nonenzymatic antioxidant levels when com-
pared to lung cancer bearing animals.
The protective role of piperine was examined during experimental lung
carcinogenesis with reference to its effect on DNA damage and the detoxi-
fication enzyme system.67 The activities of detoxifying enzymes such as
glutathione transferase, quinone reductase and UDP-glucuronosyl trans-
ferase were found to be decreased while the hydrogen peroxide level was
increased in the lung cancer bearing animals. Supplementation of piperine
(50 mg/kg) enhanced these detoxification enzymes and reduced DNA dam-
age. These results explain the understanding of association between the
anti-peroxidative effect of piperine and ultimately the capability of piperine
to prevent cancer. A significant suppression in the micronuclei formation
induced by B(α)p and cyclophosphamide following oral administration of
piperine at doses of 25, 50 and 75 mg/kg in mice has been reported.68
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Piperine has been evidenced to show chemopreventive effects when
administered orally on lung cancer bearing animals.69 The beneficial
effect of piperine is primarily exerted during the initiation phase and post-
initiation stage of B(α)p-induced lung carcinogenesis via beneficial
modulation of lipid peroxidation and membrane-bound ATPase enzymes.
The ability of piperine to prevent lung carcinogenesis induced by B(α)p
in mice and its effects on cell proliferation has been studied.70
Administration of piperine significantly decreased the levels of lipid per-
oxidation, protein carbonyls, nucleic acid content and polyamine
synthesis that were found to be increased in lung cancer bearing animals.
Piperine could effectively inhibit B(α)p-induced lung carcinogenesis in
albino mice by offering protection from protein damage and also by sup-
pressing cell proliferation. Dietary black pepper (0.5% in the diet for 15
wks) has been evidenced to suppress colon carcinogensis induced by the
procarcinogen 1,2-dimethylhydrazine (15 s.c. injections of 20 mg/kg at
weekly intervals) in rats.71
Other Physiological Effects
Animal studies
Deleterious effect of piperine on the reproductive system: Black pepper is
used as a contraceptive in folk medicine. The reproductive toxicity of
piperine has been studied in albino mice with respect to the effect on
estrous cycle, mating behavior, toxicity to male germ cells, fertilization,
implantation and growth of pups.72 Piperine (10 and 20 mg/kg body
weight) increased the period of the diestrous phase resulting in decreased
mating performance and fertility. Post-partum litter growth was not
affected by the piperine treatment and sperm shape abnormalities were not
induced at doses up to 75 mg/kg. Considerable anti-implantation activity
was recorded after 5 days post-mating oral treatment with piperine. These
results show that piperine interferes with several crucial reproductive
events in a mammalian model. The effect of piperine on the fertilization
of eggs with sperm has been investigated in female hamsters intragastri-
cally treated with piperine at doses of 50 or 100 mg/kg body weight from
day 1 through day 4 of the estrous cycle.73 During piperine treatment,
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these females were superovulated and artificially inseminated (AI) with
spermatozoa from untreated male hamsters at 12 hrs after hCG injection.
Administration of piperine to the superovulated animals markedly
enhanced the percent fertilization at 9 hrs after AI.
Piperine administered to mature male albino rats at 10 mg/kg body
weight p.o. for 30 days caused a significant reduction in the weights of
testis and accessory sex organs.74 Histological studies revealed that piper-
ine caused severe damage to the seminiferous tubule, a decrease in
seminiferous tubular and Leydig cell nuclear diameter, and desquamation
of spermatocytes and spermatids. The effect of piperine on the fertilizing
ability of hamster sperm has been investigated in vitro.75 Addition of
0.18–1.05 mM piperine reduced both the percentage of eggs fertilized and
the degree of polyspermia in a dose-dependent manner. The effect of
piperine on the epididymal antioxidant system of adult male rats has been
studied. Rats orally administered piperine at doses of 1, 10 and 100 mg/kg
body weight each day for 30 consecutive days showed a decrease in the
activity of antioxidant enzymes and sialic acid levels in the epididymis
and thereby increased reactive oxygen species levels that could damage
the epididymal environment and sperm function.76
Anti-inflammatory activity: The anti-inflammatory activity of piperine
has been reported in rats employing different experimental models
like carrageenan-induced rat paw edema, cotton pellet granuloma, and
croton oil-induced granuloma pouch.77 Piperine acted significantly on
early acute changes in inflammatory processes and chronic granulative
changes. The pungent principles of dietary spices including piperine have
been reported to induce a warming action via adrenal catecholamine
secretion.78
Hepatoprotective activity: Piperine has been evaluated for its antihepato-
toxic potential in order to validate its use in traditional therapeutic
formulations.79 It exerted a significant protection against t-butyl hydroper-
oxide and carbon tetrachloride induced hepatotoxicity by reducing lipid
peroxidation, leakage of enzymes alanine aminotransferase and alkaline
phosphatase, and by preventing the depletion of glutathione and total thi-
ols in the intoxicated mice.
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Neuropharmacological activity: To understand the effect of piperine on
the central nervous system, the neuropharmacological activity of piper-
ine administered Wistar rats (5, 10 and 20 mg/kg body weight once
daily) were determined after single, 1, 2, 3 and 4 wks of treatment.80
Piperine at all dosages examined in this study possessed antidepressant-
like activity and cognitive enhancing effects at all treatment durations,
suggesting that piperine could be a potential functional food to improve
brain function. The antidepressant-like effects of piperine and its deriv-
ative antiepilepsirine were investigated in two depressive models: the
forced swimming test and the tail suspension test.81 To further explore
the mechanisms underlying their antidepressant-like activities, the brain
monoamine levels and monoamine oxidase A and B activities were also
determined. The results indicated that after 2 wks of chronic adminis-
tration, these compounds at doses of 10–20 mg/kg significantly reduced
the duration of immobility in both models. The study demonstrated that
the antidepressant-like effects of piperine and antiepilepsirine might
depend on the augmentation of the neurotransmitter synthesis or the
reduction of the neurotransmitter reuptake. The antidepressant proper-
ties of piperine were supposed to be mediated via the regulation of
serotonergic system.
In vitro studies
Melanocyte stimulation: Melanocyte proliferation stimulants are of inter-
est as potential treatments for the depigmentary skin disorder vitiligo.
P. nigrum contains several amides with an ability to stimulate melanocyte
proliferation. It has been suggested that the methylenedioxyphenyl func-
tion is essential for melanocyte stimulatory activity.82 Black pepper water
extract and piperine promote melanocyte proliferation in vitro. Black pep-
per extract was found to possess growth-stimulatory activity in cultured
melanocytes.83 Its aqueous extract at 0.1 mg/ml was observed to cause
nearly 300% stimulation of the growth of a cultured mouse melanocyte
line, in 8 days. Hence, it is inferred that piperine is a potential repigment-
ing agent for the treatment of vitiligo. This finding supports the traditional
use of P. nigrum extracts in vitiligo and provides new lead compounds for
drug development for this disease.
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The in vitro effects of piperine on three bioenergetic reactions,
namely, oxidative phosphorylation, ATPase activity and calcium transport
by isolated rat liver mitochondria, have been investigated.84 The study
suggested that piperine inhibits mitochondrial oxidative phosphorylation
at the level of the respiratory chain. Piperine did not inhibit the mito-
chondrial ATPase activity induced by dinitrophenol and was found to
diminish calcium uptake. The influence of piperine on the enzymes and
bioenergetic functions in isolated rat liver mitochondria and hepatocytes
has been studied, and it was observed that piperine produces concentration-
related, site-specific effects on mitochondrial bioenergetics and enzymes
of energy metabolism.85
Clinical trials: Piper longum and Piper nigrum are conventionally used as
immuno-enhancers in the Indian system of traditional medicine. The
underlying mechanism, however, remains unknown.
Pepper has been used in China as a folk remedy for epilepsy. Piperine
has been identified by researchers as having anticonvulsant effects in ani-
mal models, and antiepilepsirine, a derivative of piperine, has been used
in China to treat epilepsy since 1975. A recent clinical trial on epileptic
children tested antiepilepsirine (10 mg/kg body weight; two or three times
a day) in a randomized, placebo-controlled, cross-over, double-blind trial
decreased the number of seizures in the majority of subjects.9
Black pepper’s irritant action on the respiratory tract has been har-
nessed to ease smoking withdrawal. Inhalation of black pepper essential
oil was shown to stimulate sensory signals that promoted greater smoking
cessation by decreasing withdrawal symptoms more than breathing in air
or mint/menthol.86 A clinical study has also evidenced the remarkable
effects of black pepper aromatherapy (inhalation of black pepper oil) on
dysphagia, or the difficulty to swallow, in the elderly who are at risk of
developing pneumonia, the beneficial effect being mediated by an
increase in serum levels of substance P (a neuropeptide).87
MOLECULAR TARGETS
The principal bioactive constituent of both black pepper (Piper nigrum)
and long pepper (Piper longum), the ingredients of Trikatu, which in turn
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is a constituent of many medications in the ancient systems of medicine,
has now been established as piperine. The mode of action of this alkaloid
in various medicinal effects is undoubtedly its bioavailability enhancing
influence on various structurally and therapeutically diverse drugs.
Piperine’s potential to increase the bioavailability of drugs when pre-
treated or coadministered is of great clinical significance. Clinical trials
have established that piperine increases circulatory levels of drugs such as
phenytoin (an epileptic treatment), propranolol (used for hypertension),
rifampicin (a tuberculosis medication), theophylline (lung medication),
and curcumin (a spice compound having cancer preventive and suppres-
sive potential, besides several other medicinal effects). This observed
drug bioavailability enhancing effect is due to the inhibitory interaction of
piperine with cytochrome P450 enzymes of the liver and small intestine
that are involved in drug metabolism: CYP1A2, CYP1A1, CYP2D6,
CYP3A4 and P-glycoprotein.38 Since piperine inhibits both P-glycopro-
tein and CYP3A4 expressed in intestinal enterocytes and hepatocytes, it
contributes to a major extent to first-pass elimination of many drugs.38
Piperine displays antipyretic, analgesic and anti-inflammatory activi-
ties. In the process of identifying non-steroidal anti-inflammatory
molecules from natural sources, it has been demonstrated that piperine
inhibits adhesion of neutrophils to the endothelial monolayer.88 The inhi-
bition of adhesion of neutrophils to the endothelial monolayer by piperine
is due to its ability to block the tumor necrosis factor-alpha (TNF-α)
induced expression of cell adhesion molecules, i.e. ICAM-1 (intercellular
adhesion molecule-1), VCAM-1 (vascular cell adhesion molecule-1) and
E-selectin. As nuclear factor-kappaB (NF-κB) is known to control the
transcriptional regulation of cell adhesion molecules, the effect of piper-
ine on NF-κB in the cytoplasm and in the nucleus of endothelial cells was
measured. It was observed that pretreatment of endothelial cells with
piperine blocks the nuclear translocation and activation of NF-κB by
blocking the phosphorylation and degradation of its inhibitory protein,
I-kBα. Piperine blocks the phosphorylation and degradation of I-kBαby
attenuating TNF-αinduced IkB kinase activity. These results suggest a
possible mechanism of the anti-inflammatory activity of piperine.
A current area of basic research is the activity of piperine as a TRPV1
vanilloid agonist, more powerful than the capsaicin found in chili peppers,
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to treat gastrointestinal disorders such as irritable bowel syndrome and
diarrhea, as well as chronic breast pain and urinary incontinence.89 Since
piperine has been used to stimulate the gastrointestinal tract, it could be
helpful for conditions such as diarrhea and irritable bowel syndrome,
which are not easily managed by standard care.
ABSORPTION AND METABOLISM OF PIPERINE
Animal studies: When piperine was administered to male albino rats at a
dose of 170 mg/kg by gavage or 85 mg/kg i.p., about 97% was absorbed
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Black Pepper and Its Bioactive Compound, Piperine 27
Table 3. Modulation of bioavailability of drugs, phytochemicals, and carcinogens by
black pepper and piperine.
System Remarks Reference
Humans a) Increased bioavailability of vasicine and sparteine as 39
a result of Piper longum/piperine treatment
b) Enhanced systemic availability of propranolol and 40
theophylline as a result of piperine treatment
c) Increased serum concentration of curcumin by 41
concomitant administration of piperine
d) Increased plasma levels of coenzyme Q10 by 42
coadministration of piperine
e) Increased plasma concentration of phenytoin when 43
coadministered along with piperine
f) Increased plasma concentration of antiretroviral agent 44
nevirapine when coadministered along with piperine
Rats a) Decreased metabolic activation of fungal toxin aflatoxin 48
B1and hence its increased accumulation in plasma
b) Enhanced bioavailability of β-lactam antibiotics 47
amoxicillin trihydrate cefotaxime by coadministration
of piperine
Mice a) Delayed elimination of anti-epileptic drug phenytoin by 46
treatment of piperine
b) Increased plasma levels and delayed excretion of 45
epigallocatechin-3-gallate from green tea as a result
of intragastric cotreatment with piperine
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28 K. Srinivasan
Table 4. Antioxidant, antimutagenic and cancer preventive effects of piperine.
System Remarks Reference
Antioxidant influence of black pepper and piperine
In vitro a) Inhibition/quenching of super oxides and hydroxyl 49
radicals by piperine; inhibition of lipid peroxidation
b) Marginal inhibitory effect of piperine on ascorbate- 50
Fe++-induced lipid peroxidation in rat liver microsome
c) Water and ethanol extract of black pepper exhibits 51
strong total anti-oxidant activity and inhibits
peroxidation of linoleic acid emulsion
d) Piperine protects Cu++-induced lipid peroxidation of 52
human LDL
e) Black pepper aqueous extract and piperine inhibit 53
human PMNL 5-lipoxygenase
Rats a) Piperine treatment protects against oxidative stress 55
induced in intestinal lumen by carcinogens
Streptozotocin- a) i.p. administration of piperine for 2 wks partially 54
diabetic protects against diabetes-induced oxidative stress
rats
High-fat a) Dietary black pepper/piperine reduces high-fat diet- 57
fed rats induced oxidative stress by lowering lipid peroxidation,
restoring activities of anti-oxidant enzymes and GSH
Mice a) Piperine treatment decreases mitochondrial lipid 56
peroxidation and augmented antioxidant defense system
during benzo(α)pyrene-induced lung carcinogenesis
Antimutagenic and tumor inhibitory effects
In vitro a) Black pepper is effective in reducing mutational 58
and cell events induced by procarcinogen ethylcarbamate in
lines Drosophila
b) Piperine markedly reduces the AFB1-induced formation 60
of micro-nuclei in H4IIE cells in a concentration-
dependent manner
c) Piperine counteracts CYP450 2B1 mediated toxicity 61
of AFB1in Chinese hamster cells and therefore has
chemopreventive effects against procarcinogens
activated by CYP450 2B1
(Continued)
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irrespective of the mode of dosing.90 Three percent of the administered
dose was excreted as piperine in the feces, while it was not detectable in
urine. When everted sacs of rat intestines were incubated with 100–1000 µg
of piperine, about 44-63% of the added piperine disappeared from the
mucosal side.90,91 Absorption of piperine, which was maximum at 800 µg
per 10 ml, was about 63%. The absolute amounts of piperine absorbed in
this in vitro system exceeded the amounts of other structurally closer spice
compounds such as curcumin.90 The absorbed piperine could be traced in
both the serosal fluid and in the intestinal tissue, indicating that piperine
did not undergo any metabolic change during the process of absorption.
When piperine was associated with mixed micelles, its in vitro intestinal
absorption was relatively higher. Piperine absorption in the everted intes-
tinal sac significantly increased when the same was present in micelles.91
Examination of the passage of piperine through the gut indicated that the
highest concentration in stomach and small intestine was attained at about
6 hrs. Only traces of piperine were detected in serum, kidney and spleen from
30 mins to 24 hrs. About 1–2.5% of the intraperitoneally administered piper-
ine was detected in the liver during 0.5–6 hrs after administration as
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Black Pepper and Its Bioactive Compound, Piperine 29
Table 4. (Continued)
System Remarks Reference
Rats a) Piperine administration effectively reduces 62
cyclophosphamide-induced chromosomal aberrations
in bone marrow cells
b) Dietary black pepper was evidenced to suppress colon 71
carcinogensis induced by the procarcinogen
1,2-dimethylhydrazine
Mice a) Tumor inhibitory activity of black pepper in mice 64
implanted with Ehrlich ascites tumor
b) Piperine inhibits tumor development in mice induced 59
with Dalton’s lymphoma cells and increases the
lifespan of afflicted mice
c) Antimetastatic activity of piperine on lung metastasis 65
induced by melanoma cells
d) Chemopreventive effect of piperine on 66,69,70
benzo(α)pyrene-induced experimental lung cancer
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30 K. Srinivasan
Table 5. Other biological effects of black pepper and piperine.
System Remarks Reference
Effect on reproductive system
In vitro a) Piperine decreases fertilizing ability of hamster sperm and 75
degree of polyspermia in vitro
Rats b) Continued oral intake of piperine produces reduction in 74
weights of testis, fall in sperm concentration, and decrease
in intra-testicular testosterone
Mice c) Oral intake of piperine decreases fertility due to 72
interference with crucial reproductive events in albino
mice
Anti-inflammatory activity
Rats a) Anti-inflammatory activity of piperine in experimental 77
models: carrageenan-induced rat paw edema, cotton pellet
granuloma, croton oil-induced granuloma pouch
Hepatoprotective activity
Mice a) Piperine exerted protection against t-butyl hydroperoxide 79
and carbon tetra-chloride in hepatotoxicity by reducing
lipid peroxidation
Melanocyte stimulation
In vitro a) Growth stimulatory activity of black pepper extract in 83
cultured melanocytes
Neuropharmacological activity
Rats a) Piperine administered animals possess antidepressant-like 80
activity and experience a cognitive enhancing effect
b) Antidepressant-like effects of chronically administered 81
piperine depend on the augmentation of the
neurotransmitter synthesis
Anticonvulsant effects
Humans a) Piperine treatment reduces the number of seizures in 9
epileptic children
Amelioration of dysphagia
Humans Inhalation of black pepper essential oil has remarkable 87
effects on swallowing dysfunction in patients suffering
from dysphagia
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compared with 0.1–0.25% of the orally administered dose. The increased
excretion of conjugated uronic acids, conjugated sulphates and phenols indi-
cated that scission of the methylenedioxy group of piperine, glucuronidation
and sulphation appear to be the major steps in the disposition of piperine in
the rat. After oral administration of piperine (170 mg/kg) to rats, the metabo-
lites in urine (0–96 hrs) were identified to be piperonylic acid, piperonyl
alcohol, and piperonal and vanillic acid in the free form, whereas only
piperic acid was detected in bile (0–6 hrs).92 The kidney appears to be the
major excretion route for piperine metabolites in rats as no metabolite could
be detected in feces. In a recent investigation,93 to further study the reported
differences in its metabolism in rats and humans, a new major urinary
metabolite was detected in rat urine and plasma using HPLC and character-
ized as 5-(3,4-methylenedioxy phenyl)-2,4-pentadienoic acid-N-(3-yl
propionic acid)-amide. This metabolite has a unique structure in that it
retains the methylenedioxy ring and conjugated double bonds while the
piperidine ring is modified to form the propionic acid group.
The absorption dynamics of piperine in intestine on oral absorption has
been studied.94 Using intestinal everted sacs and cycloheximide treatment
and exclusion of Na+salts from incubating medium as variables, absorp-
tion half-life, absorption rate, absorption clearance and apparent
permeability coefficient were computed. The data suggested that piperine
is absorbed very fast across the intestinal barrier, possibly acting as an
apolar molecule and forming an apolar complex with drugs and solutes. It
may modulate membrane dynamics due to its easy partitioning, thus help-
ing in efficient permeability across the barrier.
Being essentially water insoluble, piperine is presumed to be assisted
by serum albumin for its transport in blood after its intestinal absorption.
The binding of piperine to serum albumin has been examined by employ-
ing steady-state and time-resolved fluorescence techniques.95 The
binding constant for the interaction of piperine with human serum albu-
min, which was invariant with temperature in the range of 17–47°C, was
found to be 0.5 ×105M−1, having stoichiometry of 1:1. Steady-state and
time-resolved fluorescence measurements suggested the binding of
piperine to the subdomain-IB of serum albumin. These observations are
significant in understanding the transport of piperine in blood under
physiological conditions.
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Black Pepper and Its Bioactive Compound, Piperine 31
b734_BlackPepper.qxd 1/2/2009 2:53 PM Page 31
CONCLUSIONS
Black pepper or its bioactive compound piperine, the ingredients used in a
number of ancient and folk medicines, has now been demonstrated by a
number of independent investigators to possess diverse beneficial physio-
logical effects. The most far-reaching attribute of piperine is its inhibitory
influence on the hepatic, pulmonary and intestinal drug metabolizing sys-
tems. It strongly inhibits a particular cytochrome P450 and hence phase-I
reactions mediated by the same, especially aromatic hydroxylation. It also
strongly retards glucuronidation reactions of phase-II. As a result of inter-
ference with crucial drug metabolizing reactions in the liver, piperine
enhances the bioavailability of therapeutic drugs, i.e. it increases their
plasma half-life and delays their excretion. This particular inhibitory effect
of piperine on drug metabolism and hence on drug bioavailability may be
harnessed for increasing therapeutic effects. Most of the clinical studies on
piperine have focused on its effect on drug metabolism. The gastrointesti-
nal system is affected by black pepper and piperine in many ways. Both
black pepper and piperine have been evidenced to have antidiarrheal prop-
erties and a definite effect on intestinal motility and on the ultrastructure of
intestinal microvilli improving the absorbability of nutrients. Piperine has
been evidenced to protect against oxidative damage by inhibiting or
quenching free radicals as well as lower lipid peroxidation and beneficially
influence cellular antioxidant status in different situations of oxidative
stress. Piperine also possesses cytoprotective effects by retarding the acti-
vation of certain procarcinogens by the drug metabolizing system. The
antimutagenic and anti-tumor properties of piperine have been evidenced
in a few animal and cell-line studies. Among other physiological effects
piperine exerts, its potential antifertililty influence on the reproductive
system has been clearly established in in vitro and animal systems.
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