Medicine in Drug Discovery 7 (2020) 100027

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Medicine in Drug Discovery

journal homepage: https://www.journals.elsevier.com/medicine-in-drug-discovery

Review Article

Piperine: A comprehensive review of methods of isolation, purification,

and biological properties
Anshuly Tiwari, Kakasaheb R. Mahadik, Satish Y. Gabhe
⁎
Department of Pharmaceutical Chemistry, Poona College of Pharmacy, Bharati Vidyapeeth (Deemed to be University), Erandwane, Pune, 411038, Maharashtra, India

A R T I C L E I N F O A B S T R A C T

Article history: Black pepper is an autoicous and decorous vine cultivated and harvested in tropical regions of Sri Lanka and India.
Received 29 August 2019 Black pepper is one of the most commonly consumed spices, and its pungency is due to the presence of an alkaloid
Received in revised form 6 March 2020 known as piperine, volatile chemical constituents, and essential oils. Piperine is found in black pepper (piper nigrum),
Accepted 21 March 2020
white pepper, and long pepper (piper longum) belonging to the family Piperaceae. Piperine represents diverse biological
Available online 1 April 2020
activities, such as anti-inflammatory, anticancer, antiviral, anti-larvicidal, pesticide, anti-Alzheimer’s, antidepressant,
Keywords:
and most importantly piperine is known as the bioavailability enhancer. The current review article aims to explore the
Black Pepper extraction of piperine from different species of pepper, its total synthesis, pharmacokinetic study and various biological
Piperine activities of piperine. Furthermore, different combinations of piperine with available marketed drugs to improve the
Extraction and Isolation bioavailability have also been included. Different studies on piperine suggest that piperine acts as an excellent
Bioenhancer bioenhancer to improve the bioavailability of drugs with poor ADMET properties. Some studies also indicate that pip-
Total Synthesis erine shows synergistic effects when taken in combination with various classes of drugs.
© 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://
creativecommons.org/licenses/by-nc-nd/4.0/).

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Extraction Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1. Ionic liquid (IL) based ultrasonication-assisted extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.2. Maceration of black pepper with glacial acetic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.3. Extraction and Activity-Guided Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.4. Soxhlet and modified Soxhlet extractions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.5. Extraction using ethanol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.6. Extraction with Glacial Acetic Acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.7. Microwave-assisted extraction of piperine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.8. Extraction of piperine using Naviglio Extractor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.9. Micellar Extraction of piperine by Ionic Liquid (IL) Extraction Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.10. Microwave reflux extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.11. Extraction of piperine using ultrasound . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.12. Supercritical Fluid extraction technique (SFE). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3. Total Synthesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4. Biological Activities of Piperine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.1. Piperine as anti-allergic agent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.2. Effect of piperine on brain cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.3. Effect of piperine on metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
4.4. Effect of piperine as a potential bioavailability enhancer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

⁎ Corresponding author at: Department of Pharmaceutical Chemistry Poona College of Pharmacy, Bharati Vidyapeeth (Deemed to be University), Erandwane, Pune, 411038, Maharashtra, India.
Tel.: +91 9766592664.
E-mail address: satishgabhe@gmail.com. (S.Y. Gabhe).

http://dx.doi.org/10.1016/j.medidd.2020.100027
2590-0986/© 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.
0/).
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

4.5. Effect of Piperine on the Pharmacokinetics of Human Volunteers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

4.6. Synthesis and biological activity of piperic acid derivatives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
4.7. Piperine derivatives as an antineoplastic agent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
4.8. Piperine derivatives as anti-tubercular agent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5. Formulation Technologies and enhancement of therapeutic impacts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
6. Role of Pepper Species in Traditional Medicine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
6.1. Ayurveda . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
6.2. Unani system of medicine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
6.3. Siddha System of Medicine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
6.4. Marketed Formulation of piperine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
7. Conclusion and Future Prospect. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Declaration of interest statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

1. Introduction Piperine is used in traditional therapies of Chinese as well as in Indian

Among all spices, black pepper is well known as a distinctive spice
worldwide. It is also known as the ‘King of spices.’ It has a distinctive
pungent flavor due to the presence of an alkaloid piperine, along with
volatile oils, and essential oils. The content of piperine varies from
plant to plant belonging to the Piperaceae family and varies from 2%
to 7.4% in vines of black and white pepper (piper nigrum). However,
certain reports suggest a higher content of piperine up to 9% in black
pepper and 4-5% in long pepper (piper longum) [1]. The amount of pip-
erine content can be influenced by modifications in conditions of cultiva-
tion such as climate or drying conditions and the place of origin [2].
Piperine, the most abundant pungent principle present in black pepper, was
initially isolated in 1819 by Hans Christian Ørsted. He extracted a yellow crys-
talline material having molecular formula C17H19NO3 with a melting point of
128-130°C. Later, the chemical structure was elucidated, and its IUPAC name
is (2E, 4E)-5-(benzo[d] [1,3]dioxol-5-yl)1-(piperidin-1-yl)penta-2,4-dien-1-
one. Piperine is weakly basic in nature, which on hydrolysis (acidic/basic),
can be converted to piperic acid and piperidine [3]. A conjugated aliphatic
chain acts as a bridging connective structure between piperidine and 5-(3,4-
methylenedioxyphenyl) moiety. This makes piperine a unique and excellent
molecule to offer optimum attributes for the tendency of the molecule to
bind successfully to the CYP- 450 enzymes Fig. 1.
Black pepper contains four isomeric forms of piperine, namely
trans-trans isomer (1, piperine), cis-trans isomer (2, isopiperine), cis-cis
isomer (chavicine, 3), and trans- cis isomer (isochavicine, 4) (Fig. 2).
The geometrical isomers of piperine have no pungency compared to
piperine [4]. Later investigations on piperine have led to the identifica-
tion of some other alkaloids present in black pepper extract such as
piperanine, 5; piperettine, 6; piperylin A, 7; piperolein B, 8; and
pipericine, 9 [Fig. 2. and Fig. 3.] [5]. Isomerization increases with an
increase in light intensity and time exposure. Light-induced isomeriza-
tion can be seen in piperine, which can be converted into isopiperine 2,
chavicine 3, and isochavicine 4. Transformation of chavicine to piper-
ine can be seen on storage that slowly and spontaneously leads to the
loss of pungency [6]. (See Figs. 4 and 5.)
medicine. Piperine can be used widely in pain management, chills, rheuma-
tism arthritis, influenza, and fever [7]. Piperine is reported to be used for
the enhancement of blood circulation, salivation, and stimulation of appe-
tite [8]. Piperine has a multifaceted biological profile that includes pain
management [9], hypotension, vascular cell modulation [10], and anti-
cancer activity [12]. It also acts on many enzyme systems (including p-gly-
coproteins) [13,14]. Piperine has shown various biological activities such
as anti-infective, antimicrobial, insecticidal, anti-inflammatory,
antiamoebic, antiulcer, and antidepressant [16–25].
Piperine increases the absorption and bioavailability of various drug mol-
ecules [26–28]. Rajendra Prasad et al. reported the role of piperine as a poten-
tial bioavailability enhancer of drugs used in the management of tuberculosis.
Moreover, there are shreds of evidences that support the fact that piperine
possesses various pharmacological activities modulating transporter and met-
abolic enzyme activities. Moreover, it is suggested that piperine can be used
as an alternative medicine [29]. Park et al. reported the effect of piperine
on 3T3-L1 cell lines by inhibiting the expression of PPAR-γ, thus used in the
treatment of diseases related to obesity [30]. Ahmad et al. discussed the bio-
logical values and diverse biological activities of piper nigrum, effect of piper-
ine in the process of digestion, antioxidant properties, and the role in the
management of various disorders [31–33].
Bioenhancers are agents capable of increasing the bioavailability when
combined with a particular therapeutic agent without exerting any of its bio-
logical activity at the used dose. The term Bioenhancer/biopotentiator was
first defined by an Indian scientist Dr. C.K. Atal at Regional Research Labora-
tory (RRL, Jammu) currently known as the Indian Institute of Integrative
Medicine, Jammu, India. The mechanism of action of various herbal
bioenhancers can be similar or different. Bioenhancers are capable of increas-
ing the absorption from the gastrointestinal tract or inhibiting enzymes in-
volved in the biotransformation of the drug by preventing the drug
transformation to metabolites and by decreasing the rate of elimination [34].

Fig. 1. Structure of piperine. Fig. 2. Structural isomers of piperine.

2
A. Tiwari et al.
Fig. 3. Alkaloid present in black pepper other than piperine.
2. Extraction Methods
Long pepper (piper longum) kernels contain 1% - 2% piperine while in
black pepper and white pepper (piper nigrum) kernels, the amount of piper-
ine varies from 1%to 5%. There are different methods used to isolate piper-
ine. The dark brown oily substance obtained with polar organic solvents by
maceration and/or Soxhlet extraction of black pepper yields piperine and
similar amides by using the chromatographic technique. Supercritical
fluid extraction, followed by crystallization, has been used to obtain pure
piperine. Diverse methods are available in the literature to obtain piperine
from black pepper by extraction methods [Table 1].
2.1. Ionic liquid (IL) based ultrasonication-assisted extraction
Cao X et al. [35], and Sun X et al. [36] reported a technique for the ex-
traction of piperine. In this technique, an instrument called ultrasonic
water bath (Model KQ-100 DA and KQ-500, China) was used. In the tech-
nique, dried powder of black pepper (1.0 g) was mixed with different
ionic liquid (IL) solutions (10 ml) followed by the extraction of suspension
by using ultrasonic extraction. IL preparation was done by dissolving
Fig. 4. Active piperine derivatives as MAO-A inhibitor.
3
Medicine in Drug Discovery 7 (2020) 100027
different series of 1- alkyl-3-methylimidazolium ionic liquids in deionized
water in the concentration of 0.530 mol/L. The extraction method was op-
timized; the obtained suspension was subjected to ultrasound treatment
and was cooled to room temperature. The cold solution was then filtered
by using the Buchner funnel, and this filtrate was diluted up to 50 ml by
using deionized water and passed through a 0.2 mm filter. It was then sub-
jected to Ultra Performance Liquid Chromatography analysis.
2.2. Maceration of black pepper with glacial acetic acid
Shingate P et al., [37], Kanaki N et al., [38] described another method of
extraction and isolation of piperine. In this method, black pepper seeds
were ground to obtain the powder, the powdered black pepper (25 g); gla-
cial acetic acid (50 m) was taken and extracted by cold maceration process
for 5 min. This process was repeated 4 to 5 times to get better yield. The ob-
tained extracts were filtered and pooled. The pooled acetic acid solution
was further diluted with an equivalent quantity of distilled water and was
extracted with chloroform (100 ml). The extract was thoroughly washed
with a 10% NaHCO3 solution followed by 4-5 washings with water. The
chloroform layer was dried over sodium sulfate, concentrated to dryness
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Fig. 5. Metabolites of piperine.

at 60°C on a water bath. The obtained residue was then recrystallized using
chloroform by adding diethyl ether to the chloroform solution. The needle-
shaped crystals of piperine (yield: 1.12 g) were obtained. The repeated re-
crystallization process increased the purity of the crude product.
2.3. Extraction and Activity-Guided Isolation
mixture was refluxed for 20 min, and the extract was allowed to cool for 5
min. The slurry obtained was filtered by suction. This slurry was washed
with 5 ml of methylene chloride. The crude product was further recrystal-
lized using ether after providing washings with alkali to obtain the
needle-shaped crystals of piperine.

Raman G et al., [39] used piper longum fruits (3 kg) and powdered them.
This powder was extracted using 95% ethanol at 28-30°C for at least 4 times
to obtain a dark brown residue (450 g, 91.0%). This ethanolic extract was
suspended in water and partitioned between n-hexane, chloroform, ethyl
acetate, and water, respectively. The chloroform (120 ml) layer obtained
was subjected to column chromatography over silica gel with a gradient
of methanol in chloroform as a mobile phase (1, 2, 5, 10, 20, 50, and
100% each). This material was further recrystallized using chloroform
and methanol. The seeds of black pepper (5 g) were grounded, and dichlo-
romethane 10 ml was taken into a 50 ml flask with a fitted condenser. The
2.4. Soxhlet and modified Soxhlet extractions
Subramanian R et al., [40] Hamrapurkar P. D. et al. [41], and Kolhe S. R.
et al. [ [42] reported an extraction method using a Double Bypass Soxhlet
Apparatus (DBSA). In this method, a portion of plant material (40 g) was ex-
haustively extracted with methanol (250 ml) by using the Soxhlet assem-
bly. The extraction was continued for 22-24 h. The acquired extract was
then concentrated on a water bath, and the final yield of the extracted com-
pound was calculated. A 40 g sample was then introduced into a close-
capped DBSA that is connected to 2 distillation flasks, respectively through

Table 1
Methods for Extraction of Piperine.
0Sr. Extraction Method Extraction Yield Pros Cons References
No. Time

1. Ionic liquid (IL) based Different Ionic Liquids 1.96 % Cost Effective, sensitive, - [34,35]
ultra-sonication–assisted extraction Time: 30 min. efficient and ecofriendly
Detection method: Ultra Power technique.
Liquid Chromatography (UPLC Analysis)
2. Soxhlet and modified Soxhlet 11± 1h 4 ± 0.01% Mechanically efficient Time consuming, requirement [40,41]
extractions (DBSA) technique, inexpensive of azotropic mixtures to obtain
efficient extraction
3. Microwave-assisted extraction 2 min. 92 ± 1% Good reproducibility, min. An additional filtration assembly [45]
sample manipulation. to separate the residue
4. Super Critical Fluid Extraction (SFE) 2-5 h 81- 98% Short Processing Time, High pressure is required [53–56]
minimal solid residues
5. Ultrasound Extraction 5h 3.9% Inexpensive. Lacking in the Uniformity of [52]
the distribution in the
sonication energy.
6. Naviglio Extractor 3h 317.7 mg/ g Reproducible, less - [46,47]
time consuming
7. Microwave reflux extraction 90 min 2.058% Less time consuming, Risk of high pressure, [51]
High yield. high temperature, degradation
of the compound

4
A. Tiwari et al.
Y-shaped joints. The DBSA extraction with inverted Y-shaped joints was
performed using 40 g black pepper in 500 ml methanol for 12 h.
2.5. Extraction using ethanol
Sreevidya N et al. [43] reported the extraction of piperine from black
pepper seeds using ethanol as a solvent. Powdered black pepper (10 g)
was extracted with 150 ml (95%) ethanol by using Soxhlet extractor for 2
- 3 h. The extract was then filtered and was concentrated on a water bath
at 60°C. Furthermore, 10 ml of 10% potassium hydroxide solution was
added to the filtrate followed by continuous stirring. The residue of insolu-
ble material was separated, and the alcoholic extract was kept overnight,
which was filtered using a membrane filter. The yield of piperine was
found to be 3.2%.
2.6. Extraction with Glacial Acetic Acid
Gaikar V. G. et al. [46] reported a cold maceration procedure for
extracting black pepper (25 g) using 300 ml glacial acetic acid. The ob-
tained extract was then diluted using an equal volume of water and was
partitioned using chloroform. The extract was taken in the separating fun-
nel, and the chloroform layer was washed with sodium bicarbonate solu-
tion (10 %). The rotary evaporator was used to concentrate the extract.
After evaporation, the extract was purified by using column chromatogra-
phy using toluene: ethyl acetate (7:3) as a solvent system. The resinous im-
purities were washed with sodium hydroxide and 2-3 times with water to
remove alkali. The crude product was recrystallized using diethyl ether.
2.7. Microwave-assisted extraction of piperine
Mengal et al. [45,46] reported two different microwave-assisted
methods of extraction for the isolation of piperine. Both these techniques
were carried out using a microwave synthesizer. Whole black pepper
seeds were first air-dried and then pulverized using a high-speed mixer
and mechanically passed through sieves (mesh size 6). After that, this pow-
dered material was divided into different batches. To perform all the exper-
iment(s), particles of mesh size no. 6 and petroleum ether as the solvent was
used. The methods of extraction are as follows:
In the first technique, the raw material (black pepper seeds) were evenly
spread on a tray made up of glass with a thickness of 2-3 mm; it was kept in
a microwave oven with a capacity of 25 dm3. The powder (particle size
6) was exposed to the microwave for 2 min, then was suspended in petro-
leum ether. The experiment was carried out in borosilicate fully baffled
0.1 dm3 cylindrical reactors assembled with a turbine impeller that
consisted of six blades with a 2 cm diameter. The temperature of the bath
was kept constant. In addition, a sustained temperature was maintained
in a bath. Samples were withdrawn after every 15 min and were analyzed
for the estimation of piperine content. The speed of agitation was kept con-
stant, i.e., 1100 rpm to avoid mass transfer resistance (if any), the speed of
agitation did not show any effect on extraction beyond the agitation speed
of 1100 rpm. In the second technique, a modified microwave synthesizer
was used. The setup of the microwave synthesizer was modified by using
a glass vessel in the oven. From the liquid nitrogen trap, effluent gas was
provided for trapping the volatile material that included organic solvents.
No piperine was found in nitrogen traps. For the determination of the con-
tent and purity of piperine, the extraction vessel was removed from the
oven vessel. A 0.25 dm3 glass beaker containing 0.2-dm3 water was kept
along with the extraction vessel in the same posture in each of the experi-
ments to prevent the sample from excessive heating. A cavity for providing
a continuous sparging of nitrogen gas was placed on maintaining an inert
environment inside the vessel. The sparging was provided into the raw ma-
terial suspension in the organic solvent, and to maintain the suspension par-
ticles, the flow rate was kept constant.
5
Medicine in Drug Discovery 7 (2020) 100027
2.8. Extraction of piperine using Naviglio Extractor
Gigliarelli G et al. [47,48] reported a novel technique for processing the
extraction of piperine from the pepper fruits using an apparatus called
Naviglio Extractor®. Here, four extraction methods, namely decoction,
maceration, solid-liquid extraction, and solid-liquid dynamic extraction,
were used. The method was carried out using ethanol (96%) and the
study of chemical constituents was also carried out (yield: 78%).
2.9. Micellar Extraction of piperine by Ionic Liquid (IL) Extraction Method
Black pepper suspension was prepared in an aqueous solution (10 mg
black pepper, 990 mg IL solution, 50 mg black pepper, 950 mg IL solution,
and 100 mg black pepper, 900 mg IL solution) and was agitated at 25°C for
3 h. The supernatant sample of internal standard (60 mg phenol in 100 ml
of water, 10 μl) was diluted using 5 ml ionic liquid solution. A fraction of
the diluted solution (1 ml) was then mixed with 200 μl of internal standard,
filtered using a 2-μm syringe filter, and analyzed using HPLC [49–51].
2.10. Microwave reflux extraction
Abayomi O. O. et al. [52] reported the extraction of piperine using a do-
mestic microwave. Piper nigrum powder (5 gm) was hydrated with distilled
water. The purpose of hydrating the sample was to allow homogeneity in
the mixture. This homogenized mixture was loaded in the microwave reac-
tor and irradiated under operating conditions generated from the Taguchi
experimental array. Moreover, the microwave oven operated with the aid
of “easy-control” software, was programmed for 3-level control of the mi-
crowave power, irradiation time, and temperature. The supernatant solu-
tion was then filtered using the 0.45 μm PTFE microfilter for subsequent
spectroscopic quantification. The yield of bioactive (piperine) by using
the extraction technique under optimum conditions for black and white
pepper was 2.05% w/w and 4.27% w/w, respectively.
2.11. Extraction of piperine using ultrasound
Rathore et al. [53] reported a specific and high-yielding method for the
extraction of piperine by using the application of ultrasound-assisted ex-
traction (UAE) method with the optimal power of 125W for 5 h. Overall,
the UAE method specifically proved efficient for the optimization of a bet-
ter yielding and less time-consuming method of all other methods (yield:
5.8 mg/g).
2.12. Supercritical Fluid extraction technique (SFE)
SFE is widely accepted as a popular technique used for the extraction
and isolation of active constituents from various natural sources. The ex-
traction of piperine using SFE was carried out using optimum conditions
to provide carbon dioxide. Temperature was maintained at 1°C and the
pressure was maintained at 73.8 bar for 2-5 h, along with the lower polarity
that has a significant role in the extraction of organic compounds (yield:
81%-98%) [54,55]. Sovova et al. reported the extraction of piperine using
the same parameters with a yield of 7.6% w/w [56].
Table 2
Schemes of synthesis of piperine.
Sr. No. Schemes of Synthesis Reference
1. [58]
2. [59]
3. [60]
4. [61]
5. [62]
6. [63]
7. [64]
8. [65]
9. [66]
A. Tiwari et al.
3. Total Synthesis
The prime limitation of natural products used as medicine or as starting
material for semisynthetic chemical reactions is a fact that most of the nat-
ural products are available in minimal quantity. It needs laborious proce-
dures for extraction, isolation, and purification of the active compound.
Several synthetic procedures are reported for the synthesis of piperine
[Table 2].
Takeda et al. reported the first synthetic procedure for piperine involv-
ing the condensation of acyl chloride of piperic acid with piperidine. Tsuboi
and Takeda (1882) presented a three-step synthesis of piperine using piper-
onal, in which the propargylic alcohol and N-acetylpiperidine diethyl acetal
was condensed to give allenamides, which was then converted to a mixture
of piperine and isochavicine in the ratio of 65 : 35 [57,58].
Ladenburg et al. reported the synthesis of piperonal. It was synthesized by
using Reamer- Tiemann reaction from catechol as a starting material. Further-
more, piperine was synthesized by Claisen Schmidt condensation, which in-
volves the condensation of piperonal with acetaldehyde in the presence of a
base such as NaOH to give cinnamoyl aldehyde derivative [(E)-3-(benzo[d]
[1,3]dioxol-5-yl) acrylaldehyde]. This cinnamoyl aldehyde derivative was
further heated with sodium acetate solution (Perkin Reaction) to get piperic
acid. The piperic acid was converted to acid chloride using PCl5, and was fur-
ther treated with piperidine to yield piperine [59].
Olsen and Spessard et al. described a stereoselective two-step synthesis
of piperine. In this reaction, piperic acid was obtained by following a
vinylogous Wadsworth-Horner modified Wittig reaction of piperonal with
an anion derived from methyl (E)-4-diethylphosphono-2butenoate, which
on further methoxide-catalyzed aminolysis along with piperidine gave pip-
erine (Yield; 86%) [60].
Mandai et al. reported a reaction by coupling sulfone, synthesized from
piperonal and aldehyde to give acetate, which further underwent a double
elimination reaction to give piperine with stereo control of 90% ee. The re-
action procedure is highly stereoselective by a double-elimination reaction
of β-acetoxy sulfone with a yield of 77% [61].
Sloop et al. reported a reaction in microscale quantity involving the re-
action of methyl 2-butenoate using n-bromosuccinimide followed by aldol
condensation to give ester. It was further saponified to give piperic acid.
Aminolysis of piperic acid with piperidine led to the formation of piperine
(yield: 50%) [62].
Chandrasekhar et al. exploited a reaction between nucleophiles of car-
bon, i.e., R-MgX and R-Li over aldehyde, tosylhydrazones of aromatic and
heteroaromatic scaffold for the synthesis of piperine (yield: 72%) [63].
Schobert, R. et al. reported a synthetic method involving a reaction between
three components, namely aldehydes ketenylidene, triphenylphosphorane, and
piperidine, which led to (E)- α,β unsaturated amides (yield: 94%) [64].
Inatani R. et al. reported that the biosynthesis of piperine involves L-
lysine and cinnamoyl- CoA. Decarboxylation of L-lysine into cadaverine
that further undergoes oxidative deamination using amine oxidase contain-
ing copper to give 5-aminopentanal as an intermediate. This intermediate
further undergoes cyclization to give Δ1- piperidine Schiff base whose re-
duction gives piperidine. Simultaneously, the production of piperonal -
CoA from cinnamoyl coenzyme A takes place by chain elongation using
malonyl-CoA in a Claisen-like reaction. In the final step, pipernoyl-CoA re-
acts with piperidine to form piperine (yield: 34%) [65].
Han L. C. et al. reported that piperine could be synthesized
stereoselectively by the Horner- Wadsworth-Emmons Reaction. It involves
the reaction of phosphonate ester of methyl-4-bromo-2-butenoate and piper-
onal in the presence of sodium methoxide to form the trans alkene,
methylpiperate {(E, E)-5-(3, 4-methylenedioxyphenyl)-2,4- pentadienoate}.
Thus, the reaction of methyl piperate with piperidine in the presence of so-
dium methoxide and methanol gives piperine (yield: 70%) [66].
4. Biological Activities of Piperine
It is evident from many studies that piperine has a potential application
in drug discovery and development in diverse human ailments. Here in this
6
Medicine in Drug Discovery 7 (2020) 100027
review, an overview of the biological activities of piperine having signifi-
cant impact on the disease state on human is described.
4.1. Piperine as anti-allergic agent
Aswar et al. reported that piperine can be used as an anti-allergic agent.
It presents the allergic response through mast cell stabilization through the
inhibition of released mediators such as interleukin-6, interleukin-1b,
immunoglobulin-E, and histamine. The study on ovalbumin-induced aller-
gic rhinitis, suggests that piperine can be used as a xenobiotic agent for clin-
ical use [67]. Kim and Lee reported piperine as a major component in the
inhibition of airway inflammation using a murine model of bronchial
asthma by suppressing the Th-2 cytokines namely interleukin-4,
interleukin-5, and interleukin-13, by enhancing the expression of
Transforming Growth Factor (TGF- β) gene, and eosinophil CC chemokine
receptor (CCR3) expression. Piperine might be capable of acting as a
downregulator of immunomodulatory Th-2 cytokines [68]. Dong et al. re-
ported the effect of piperine on periodontitis by significantly preventing al-
veolar bone loss and tissue breakdown in a dose-dependent manner, the
effect of piperine that is ascribed in the inhibition of interleukin-1β and
metalloproteinase-8 and -13 (MMP-8 and MMP-13). Therefore, it can be
stated that piperine gives potent anti-inflammatory activity on the peri-
odontitis model [69]. Liu et al. reported that piperine significantly inhibits
the cyclooxygenase (COX) enzyme at the concentration of 25 μg/ml by 30%
- 80% [70]. Hu et al. (2015) reported that piperine acts as an agonist that
induces the Pregnane X Receptor (PXR), which further induces the enzyme
CYP-3A4 gene expression at protein and mRNA levels. Results supported
the finding that piperine can prevent and inhibit the activity of the colonic
inflammatory disease by CYP450 expression using the induction of the PXR
receptor [71].
Bang and Oh et al. presented evidence for piperine to be an anti-
inflammatory, antiarthritic, antiaging agent and was tested on the arthritic
model by Interleukin (IL) 1β-stimulated synoviocytes. The results of the
study showed piperine (10 ng/ml) to act in a dose-dependent manner and
can act by inhibiting the COX-2 and, more particularly, inhibiting the pro-
duction of prostaglandin E-2 (PGE-2) than IL-6 productions by ELISA and
RT-PCR analysis [72].
4.2. Effect of piperine on brain cells
Narasimhamurthy K. et al. reported that the employment of mammalian
assays was developed for the mutagenic activity. The study concluded that
neither capsaicin nor piperine was found to be an inducer for the mutage-
nicity in both somatic and germ cells [73].
D'hooge et al. reported that piperine exhibits diverse CNS activities,
namely anti-epileptic, antidepressant, and in various neurodegenerative
disorders [74]. Ren T. et al. reported that piperine, when given orally,
both in vitro and in vivo, reaches the brain effectively and shows limited he-
patic metabolism at the dose of 35 mg/kg. The inclination of piperine on
the change in the concentration of blood was parallel to that of the brain.
There is no delay in reaching the drug from plasma to the brain; the limited
transport was not observed in case of piperine in the blood-brain barrier
[75]. Yun et al. reported that piperine and piperine like alkylamides are in-
ducing agents of brain-derived neurotrophic factor-IV (BDNF) promotor in
the neuro 2α- cells. Piperylin increases the retinoic acid-induced neurite
overgrowth. Structure-activity relationship (SAR) between piperine like
alkylamide and other similar structures of piperine showed that the BDNF
inducing agent can be useful for the treatment of depression and can be a
useful therapeutic agent [76].
Osamah B. et al. reported the activity of piperine against monoamine ox-
idase (MAO-A) inhibitor. The study showed that piperine could be structur-
ally modified by replacing the piperidine ring. These compounds were
primarily tested against the MAO-B rather than MAO- A enzymes [Com-
pound 10 (MAO-B, IC50 value = 6.39 μM) and Compound 11 (MAO-B,
IC50 value = 0.49 μM]. The introduction of the oxygen atom in the struc-
ture reduced the activity. Conversion of the piperidine ring into pyrrolidine
A. Tiwari et al.
ring led to a 10-fold decrease in the activity. An increase in the number of
carbon atoms in the ring resulted in the loss of activity of Compound 12
(MAO-B, IC50 value = ≥100 μM). The results showed a slight increase in
the MAO-B inhibitory activity on the removal of the N atom in Compound
14. Nitrogen atom decreases the lipophilicity of the ring structure, and
hence can be a factor of increase in the affinity of Compound 13 (MAO-
B: IC50 = 0.666 μM). It showed more potency than Compound 14
(MAO-B: IC50 = 1.13 μM) [77].
Mori et al. reported that a dose of 60 mg/kg (intraperitoneal) of piperine
is capable of reducing or abolishing convulsions in the E-1 mice. The effec-
tive dose of 21.1 mg/kg (intraperitoneal) was administered to mice, and
after that, the estimation of the 5-HT, norepinephrine, and dopamine was
carried out. The findings suggest that the level of 5-HT was significantly
higher in cerebral cortex in mice compared to that of control mice. The in-
creased amount of 5-HT directly affects the mechanism associated with the
inhibitory activity of convulsions by treatment with piperine. Contrarily,
the effect of piperine on the hypothalamus of the brain and at the same
time, the lower concentration of norepinephrine was observed in every
part of the brain [78].
Wattanathorn et al. reported the effect of piperine on Alzheimer’s dis-
ease, neurodegeneration, and memory impairment. The report states that
piperine improved memory impairment and protects against the cognitive
neurodegeneration induced by AF64A. The mechanism was associated
with an increase in the neuronal density and inhibition of acetylcholinester-
ase enzyme in the hippocampus [79,80].
4.3. Effect of piperine on metabolism
Shrinivasan et al. reported the influence of different spices on the metab-
olism of the body [81]. Bhat and Chandrasekhar et al. reported the in vivo
ADME pathway for piperine. It was reported that piperine was absorbed
97% after oral and intraperitoneal (ip) administration. Remaining 3%
was extracted in feces. Here, there was no trace of piperine in the free
form in urine. Therefore, it was concluded that piperine was excreted in
feces following the intraperitoneal administration pathway, which may
be due to enterohepatic circulation. On the other hand, in vitro absorption
of piperine was observed in the duodenal segment other than the jejunal
and ileal segment. The added concentration of piperine 47%-64% piperine
was incubated and observed in sac tissues as well as in the serosal fluid;
moreover, 5%-10% of piperine was found in the serosal fluid alone [82].
Bhat B G. et al. came up with another report presenting the interaction of
piperine and safrole with many of the different forms of cytochrome P-
450 (CYP-450). The result revealed that piperine did not exhibit any induc-
tion of CYP- 450 enzyme, whereas safrole shows a significant induction of
the same [83].
Bhat et al. reported the metabolic pathway, biotransformation products of
piperine, and the conjugates obtained after the metabolism of piperine. In the
study of the metabolic pathway, five significant constituents were identified
along with their conjugates, namely piperic acid, piperonylic acid, piperonyl
alcohol, piperonal, and lastly vanillic acid. The absorption of piperine was ob-
served from the portal vein where piperine first undergoes hydrolysis to
piperic acid in the liver and later by the oxidation of the side chain; it is trans-
formed into piperonylic acid. It was observed that piperine could also lead to
the formation of piperonal by the oxidative cleavage of the side chain, which
was further reduced to piperonyl alcohol [84].
Badmaev et al. presented a report concerning piperine levels on
coadministrated with coenzyme Q10. It presented a practical approach
for the estimation of the enhancement of nutrient bioavailability. Piper-
ine also acted as a potent bioenhancer particularly, in patients who were
diagnosed with low plasma levels of coenzyme Q10 and in elderly pa-
tients with poor absorption from gastrointestinal tract [85]. Bhardwaj
et al. reported piperine (a principal constituent of black pepper) for
the inhibition of p-glycoprotein and enzyme CYP-3A4 [86]. Inshad Ali
Khan et al. and Brenwald N. P. et al. reported piperine to be a potential
bioenhancer of ciprofloxacin against Staphylococcus aureus [87,88].
Mona et al. reported that piperine shows anti-mutagenic activity against
7
Medicine in Drug Discovery 7 (2020) 100027
various carcinogens on sister chromatid exchanges and chromosomal
aberration in mice using mitomycin-C (MMC) as an inducing agent. In
vivo cytogenetic activity of piperine was observed using MMC as a free
radical generator and DNA-alkylating agents for the mutation of the
genotoxic enzymes [89–91]. Faik Gökalp et al. reported thermodynamic
models using the density functional theory and the Hartree Fock method
for the estimation of piperine distribution in body tissues. These
methods were optimized and the observation in terms of piperine was
made. Calculations of thermodynamic methods revealed that piperine
readily dissolves in blood and reaches to whole body tissues having a
high dipole moment. It is notably less stable; therefore, it reacts with
radical forms and in turn reduces detrimental effects. It thus prevents
damage to the cells’ structures [92].
4.4. Effect of piperine as a potential bioavailability enhancer
There are many evidences available to show that piperine can be a po-
tential bioenhancer when given in combination with xenobiotic agents.
This is due to its ability to inhibit the p glycoprotein and CYP-3A4 enzyme,
which play an essential role in the metabolism of xenobiotic agents
[Table 3]. (See Table 4.)
Bajad et al. reported that piperine could be a major factor in the inhibi-
tion of the gastric emptying of solid and/or liquid diets and the gastrointes-
tinal transit in animals (mice) in a dose- and time-dependent manner when
treated with the 1 mg/kg and 1.3 mg/kg doses [93]. Bedada et al. reported
in the study that piperine may act as a potent bioavailability enhancer by
inhibiting the p-glycoprotein-mediated efflux mechanism. The inhibition
associated with piperine and p-glycoprotein efflux may be a potential and
a beneficial therapeutic effect for the intestinal absorption of drugs or as a
potent bioenhancer for drugs that are possibly poorly absorbable [94].
Athukuri B. L. et al. reported the effect of piperine on the pharmacokinetics
of domparidone- treated animals with a dose of 20 mg/kg (oral as well as an
intraperitoneal route). The mean plasma concentration was measured by
HPLC analysis. Results suggested that there is a slight increase in mean
plasma concentration (Cmax), elimination half-life (T1/2), area under the
curve (AUC) of the domparidone when given alone or in combination
with piperine [95]. Singh A. et al. reported the effect of piperine on the
pharmacokinetic profile of rifampicin and isoniazid, alone or in combina-
tion with piperine. Results suggested that, the coadministration of rifampi-
cin with piperine results in relative bioavailability of 141.7%, and for the
isoniazid, the relative bioavailability was found to be 67.19% [96]. Hiwale
AR et al. reported the effect of piperine on the pharmacokinetics of beta-
lactam antibiotics in rat models [97].
Pattanaik et al. (2009) reported the effect of piperine on the pharma-
cokinetic profile of carbamazepine. In the study, 10 patients were
treated with carbamazepine alone (300 mg/500 mg). Similarly, these
patients were treated with piperine (20 mg p.o.), and the plasma sam-
ples were collected at similar time points. The pharmacokinetic profile
of both groups was evaluated by the student t-test method, and the bio-
availability of carbamazepine was evaluated. The possible mechanism
can be by lowering the elimination of the administered drug or by en-
hancing the absorption [98]. Pattanaik et al. studied the effect of piper-
ine administration alone or in combination with diphenylhydantoin
(DPH) (20 mg). Mean plasma concentration increased after the adminis-
tration of piperine along with DPH in both the phases namely, the elim-
ination (150 mg/200 mg) as well as in the absorption phases (150 mg/
200 mg). Results support the finding that piperine is a good
bioenhancer. The addition of piperine with DPH as a bioenhancing
agent proves the beneficial effect of piperine on the mean plasma levels
without causing the drug to accumulate [99].
Gabhe et al. (2018) reported that piperine shows significant changes in the
pharmacokinetic profile of simvastatin (10 mg/kg), verapamil (10 mg/kg),
and secnidazole (10 mg/kg) when given alone and in combination with a
fixed-dose of piperine (10 mg/kg). Piperine exhibits 2.53, 1.55, and 1.08
fold increases in the bioavailability of these drugs, respectively [100–102].
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Table 3
Pharmacokinetic activity of various drugs alone in combination with piperine
Sr. Drug Pharmacokinetic Profile Clinical/ Inference of the study Reference
No (Category) Preclinical
Drug Alone In combination with piperine
Study

1. Domperidone Route: Oral Administration Route: Oral Administration Male Wistar Reduction in the dose of domperidone due to
(D2 receptor Dose:10 mg/kg Dose: 10 mg + 20 mg Rats the inhibition of P- gp and CYP 3A1 dual [95]
antagonist) tmax: 2 h t1/2: 1 h substrate.
Cmax: 114.4±15.36 μg/ml Cmax: 178.22±13.87 μg/ml
AUC 0-t: 1122.94±114.9μg/ml/h AUC 0-t: 1489±72.00μg/ml/h
Relative Bioavailability: 141.7%
Bioavailability (Fold increase):
1.32
2. Amoxicilin Route: Oral Administration Route: Oral Administration Albino Rats Piperine may be used as an adjuvant in with the
(β- lectum Dose:100 mg/kg Dose: 100 mg + 10 mg β- lactam antibiotics by reducing the [97]
antibiotic) tmax: 1.5 ± 0.5 h Cmax: 8.0 ± 0.34 μg/ml toxicological
t1/2: 1.18 ± 0.48 h AUC 0-t: 17.80 ± 0.48 μg/ml/h profile and reduce the dosage frequency of the
Cmax: 4.2 ± 0.28 μg/ml Bioavailability (Fold increase): antibiotics.
AUC 0-t: 10.73 ± 0.50 μg/ml/h 1.65
Route: Oral Administration
Dose: 100 mg + 20 mg
Cmax: 9.4 ± 0.27 μg/ml
tmax: 1.1 ± 0.3 h
t1/2: 1.63 ± 0.67 h
AUC 0-t: 22.08 ± 0.49 μg/ml/h
Bioavailability (Fold increase):
2.05
3. Cefotaxime Route: Oral Administration Route: Oral Administration Albino Rats Piperine may be used as an adjuvant in with the
(β- lectum Dose:10 mg/kg Dose:10 mg/kg + 10 mg/ kg β- lactam antibiotics by reducing the [97]
antibiotic) tmax: 0.5 ± 0.28 h tmax: 0.55 ± 0.3 h toxicological
t1/2: 1.08 ± 0.57 h t1/2: 1.56 ±0.32 h profile and reduce the dosage frequency of the
Cmax: 20.5 ± 0.3 μg/ml Cmax: 31.0 ± 0.32 μg/ml antibiotics.
AUC 0-t: 32.64 ± 0.34 μg/ml/h AUC 0-t: 55.85 ± 0.45 μg/ml/h
Bioavailability (Fold increase):
1.72
Route: Oral Administration
Dose:10 mg/kg + 20 mg/ kg
tmax: 0.60 ± 0.31 h
t1/2: 1.78 ±0.81 h
Cmax: 35.13 ± 0.25 μg/ml
AUC 0-t: 71.09 ± 0.42 μg/ml/h
Bioavailability (Fold
increase):2.17
4. Cefadroxil Route: Oral Administration Route: Oral Administration Albino Rats Piperine may be used as an adjuvant in with the
Monohydrate Dose:100 mg/kg Dose:100 mg/kg + 10 mg/ kg β- lactam antibiotics by reducing the [97]
(β- lectum tmax: 1 ± 0.13 h tmax: 1 ± 0.13 h toxicological
antibiotic) t1/2: 2.97 ± 0.75 h t1/2: 2.85 ± 0.65 h profile and reduce the dosage frequency of the
Cmax: 45.25 ± 0.78 μg/ml Cmax: 45.5 ± 0.78 μg/ml antibiotics.
AUC 0-t: 152.925 ± 1.4 μg/ml/h AUC 0-t: 142.34 ± 1.35 μg/ml/h
Bioavailability (Fold increase):
0.93
Route: Oral Administration
Dose:100 mg/kg + 20 mg/ kg
tmax: 1 ± 0.10 h
t1/2: 2.95 ± 0.85 h
Cmax: 43.5 ± 0.93 μg/ml
AUC 0-t: 152.97 ± 1.55μg/ml/h
Bioavailability (Fold increase):
0.98
5. Carbamazepine Route: Oral Administration Route: Oral Administration Human Co-administration of piperine with CBZ
(Antiepileptic) Dose: 300 mg/kg Dose: 300 mg/kg + 20 mg/ kg Volunteers decreases the rate of elimination due to [99]
tmax: 4.10 ± 0.05 h tmax: 3.90 ± 0.37 inhibition of biotransformation and/ or
t1/2: 7.39 ± 1.09 h t1/2: 6.42 ± 1.54 h elimination of the drug.
Cmax: 6.69 ± 0.56 μg/ml Cmax: 7.21 ± 0.37 μg/ml
AUC 0-t: 66.12 ± 5.23 μg/ml/h AUC 0-t: 73.02 ± 4.51 μg/ml/h
Route: Oral Administration Bioavailability (Fold increase):
Dose: 500 mg/kg 1.10
tmax: 3.70 ± 0.59 h Route: Oral Administration
t1/2: 8.80 ± 2.01h Dose: 500 mg/kg + 20 mg/ kg
Cmax: 7.90 ± 0.5 μg/ml tmax: 3.20 ± 0.41h
AUC 0-t: 81.37 ± 4.90 μg/ml/h t1/2: 5.99 ± 0.97h
Cmax: 8.70 ± 0.45 μg/ml
AUC 0-t: 91.70 ± 5.02 μg/ml/h
Bioavailability (Fold
increase):1.12
6. Phenytoin Route: Oral Administration Route: Oral Administration Piperine enhances the bioavailability of [100,103]
(Antiepileptic) Dose: 200 mg/kg Dose: 150 mg/kg + 20 mg/ kg phenytoin at steady- state concentration in
tmax: 3.40 ± 0.51 h tmax: 3.20 ± 0.32 h epileptic patients may be due to increase in

8
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Table 3 (continued)

Sr. Drug Pharmacokinetic Profile Clinical/ Inference of the study Reference

No (Category) Preclinical
Drug Alone In combination with piperine
Study

t1/2: 14.06 ± 3.07 h t1/2: 9.02 ± 1.77 h the rate of absorption of the drug.
Cmax: 12.14 ± 1.97 μg/ml Cmax: 13.31 ± 2.18 μg/ml
AUC 0-t: 131.92 ± 19.94 μg/ml/h AUC 0-t: 143.92 ± 19.70 μg/ml/h
Route: Oral Administration Bioavailability (Fold increase):
Dose: 200 mg/kg 1.09
tmax: 3.40 ± 0.51 h Route: Oral Administration Human
t1/2: 14.06 ± 3.07 h Dose: 200 mg/kg + 20 mg/ kg Volunteers
Cmax: 12.14 ± 1.97 μg/ml tmax: 3.60 ± 0.58 h
AUC 0-t: 131.92 ± 19.94 μg/ml/h t1/2: 6.25 ± 1.2 h
Cmax: 18.24 ± 1.72 μg/ml
AUC 0-t: 192.97 ± 17.40 μg/ml/h
Bioavailability (Fold increase):
1.17
7. Fexofenidine Route: Oral Administration Route: Oral Administration Human Piperine increase the bioavailability
(antihistaminic) Dose: 120 mg/kg Dose: 120 mg/kg + 20 mg/ kg Volunteers possibility due to the inhibition of P- gp [94]
tmax: 2.4 ± 0.4 h tmax: 2.4 ± 0.4 h mediated drug efflux.
t1/2: 12.1 ± 3.2 h t1/2: 13.1 ± 1.8 h
Cmax: 406.9 ± 71.4 μg/ml Cmax: 767.0 ± 149.0μg/ml
AUC 0-t: 3403.7 ± 825.3 μg/ml/h AUC 0-t: 5724.7 ±
1253.4μg/ml/h
Bioavailability (Fold increase):
0. 54
8. Verapamil Route: Oral Administration Route: Oral Administration Male Wistar Rat -
(Calcium channel Dose: 10 mg/kg Dose: 10 mg/kg + 10 mg/ kg [100]
blocker) tmax: 0.5 h tmax: 0.5 h
t1/2: 10.66 h t1/2: 13.13 h
Cmax: 2.2456 μg/ml Cmax: 4.385 μg/ml
AUC 0the -t: 5.725 μg/ml/h AUC 0-t: 12.10 μg/ml/h
Bioavailability (Fold increase):
1.35
9. Simvastatin Route: Oral Administration Route: Oral Administration Male Wistar Rat Co- administration of piperine with simvastatin
(Antihyperlipidemic) Dose: 10 mg/kg Dose: 10 mg/kg + 10 mg/ kg increase the bioavailability of the drug whereas [102]
t1/2: 4.85 h t1/2: 6.33 h the administration of piperine derivatives
Cmax: 321 μg/ml Cmax: 703 μg/ml (2 AP, 4 AP) shows significant decrease in the
AUC 0-t: 88.787 μg/ml/h AUC 0-t: 224.55 μg/ml/h elimination of simvastatin.
Bioavailability (Fold increase):
2.53
10. Secnidazole Route: Oral Administration Route: Oral Administration Male Wistar Rat Piperine derivative when compared with
(Anti-infective) Dose: 10 mg/kg Dose: 10 mg/kg + 10 mg/kg secnidazole [101]
t1/2: 9.358 h t1/2: 10.570 h alone, there is a significant increase in plasma
Cmax: 50.292 μg/ml Cmax: 66.229 μg/ml concentration in plasma, volume of distribution
AUC 0-t: 236.361 μg/ml/h AUC 0-t: 256.886 μg/ml/h and decrease in the rate of elimination.
Fold increase: 1.08
11. Curcumin Route: Oral Administration Route: Oral Administration Rats Piperine enhances the serum concentration and
Dose: 2000 mg/kg Dose: 2000 mg/kg + 10 mg/kg bioavailability of curcumin due to increase in the [129]
tmax: 0.83 ± 0.05 h tmax: 1.29 ± 0.23h rate
t1/2: 0.31 ± 0.07 h t1/2: 0.47 ± 0.03 h of absorption and reduction in the
Cmax: 1.35 ± 0.23 μg/ml Cmax: 1.80 ± 0.16 μg/ml biotransformation
AUC 0-t: 2.36 ± 0.28 μg/ml/h AUC 0-t: 3.64 ± 0.31 μg/ml/h
Bioavailability (Fold increase):
1.55
12. Curcumin Route: Oral Administration Route: Oral Administration Human Piperine enhances the serum concentration and
Dose: 2000 mg/kg Dose: 2000 mg/kg + 10 mg/kg Volunteers bioavailability of curcumin due to increase in the [129]
tmax: 0.83 ± 0.05 h tmax: 1.29 ± 0.23h rate
Cmax: 0.006 ± 0.005 μg/ml t1/2: 0.41 ± 0.17 h of absorption and reduction in the
AUC 0-t: 0.004 μg/ml/h Cmax: 0.18 ± 0.16 μg/ml biotransformation.
AUC 0-t: 0.08 ± 0.01 μg/ml/h
Relative Bioavailability: 2000%
Bioavailability (Fold increase):
20.00
13. Gatifloxacin Route: Oral Administration Route: Oral Administration Layer Bird Piperine increase the bioavailability of
Dose: 10 mg/kg Dose: 10 mg/kg + 15 mg/kg Gatifloxacin by decreasing the permeability [130]
t1/2: 3.74 ± 0.073 h tmax: 0.5 h of epithelial cell membrane in the GIT,
Cmax: 1.74 ± 0.023 μg/ml t1/2: 4.03±0.097 h suppression of first pass metabolism, and
AUC 0-t: 15.25 ± 0.219 μg/ml/h Cmax: 2.14±0.019 μg/ml inhibition of enzyme responsible for
Relative Bioavailability: 74.52 ± AUC 0-t: 17.54 ± 0.204 μg/ml/h metabolism of Gatifloxacin in the liver.
1.021 Relative Bioavailability: 85.74 ±
0.956
14. Phenytoin Route: Oral Administration Route: Oral Administration Human -
(Antiepileptic) Dose: 300 mg Dose: 300 mg + 20 mg Volunteers [103]
t1/2: 3.0 ± 0.16 h t1/2: 4.0 ± 1.0 h
tmax: 5.14 ± 0.24 h tmax : 4.8 ±0.34 h
Cmax: 4.08± 0.21 μg/ml Cmax: 5.2 ± 0.20 μg/ml

(continued on next page)

9
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Table 3 (continued)

Sr. Drug Pharmacokinetic Profile Clinical/ Inference of the study Reference

No (Category) Preclinical
Drug Alone In combination with piperine
Study

AUC 0-t: 104.3 ± 2.77 μg/ml/h AUC 0-t: 156.26 μg/ml/h

Bioavailability (Fold increase):
1.497
15. Propranolol Route: Oral Administration Route: Oral Administration Human -
(β- adrenergic Dose: 40 mg Dose: 40 mg + 20 mg Volunteers [104]
blocker) t1/2: 9.05 h t1/2: 9.38 h
Cmax: 45.0 ng/ml Cmax: 92.0 ng/ml
AUC 0-t: 561.0 ng/ml/h AUC 0-t: 1140 ng/ml/h
Bioavailability (Fold increase):
2.03
16. Theophylline Route: Oral Administration Route: Oral Administration Human -
(anti-asthmatic) Dose: 150 mg Dose: 40 mg + 20 mg Volunteers [104]
t1/2: 6.58 h t1/2: 9.38 h
Cmax: 4.55 μg/ml Cmax: 92.0 μg/ml
AUC 0-t: 43.8 μg/ml/h AUC 0-t: 83.7 μg/ml/h
Bioavailability (Fold increase):
1.91

4.5. Effect of Piperine on the Pharmacokinetics of Human Volunteers
Strong shreds of evidences are available in the literature that represent the
effect of piperine as a bioenhancer with various diverse classes of medicinal
agents/Ayurvedic formulations, which was tested against human volunteers.
Amla et al. presented a report on the investigation of the effect of piper-
ine over the pharmacokinetic profile of antiepileptic agent phenytoin,
which has poor bioavailability. They investigated interindividual variation
of its bioefficacy. In the presented research work, five human volunteers se-
lected for the study were treated with 300 mg of phenytoin alone and in
combination with piperine with 20 mg/kg for seven days. The results ob-
tained from the study suggested that piperine when administered in
single-dose altered the pharmacokinetic profile of phenytoin by prolonging
the elimination half-life (P < 0.01), decrease in the absorption half-life (P
< 0.05), and lastly, high AUC (P < 0.05) when compared with phenytoin
administered alone. Therefore, it became evident that piperine, when ad-
ministered in multiple doses shows a more significant impact on the phar-
macokinetic profile of phenytoin [103].
Raina et al. reported the influence of a fixed dose of piperine on the bioavail-
ability of propranolol and theophylline. In the presented report, the fixed dose
of propranolol (40 mg) and theophylline (150 mg) along with piperine (20 mg)
was administered daily for seven days in the crossover study that was per-
formed on six healthy human volunteers. Plasma propranolol levels were esti-
mated using the spectrofluorimetric method, while theophylline levels in
serum was analyzed by EMIT. The study concluded that there is a significant en-
hancement of bioavailability of drugs when coadministered with piperine
[104]. Rezaee et al. presented supporting research work on the influence of pip-
erine on the plasma concentration of midazolam using healthy human

Table 4
Patents on Piperine
Sr. Patent No. Title Description of the invention Author and
No. Year

1. US5536506A Use of piperine to increase the bioavailability The patent discloses the extraction, isolation and purification method for the production of Majeed et al.
of nutritional compounds piperine. In addition, the compositions contain 98% pure piperine alkaloid in combination (1996)
with several pharmaceutical compositions to increase the bioavailability.
2. EP0810868B1 Use of piperine as a gastrointestinal The patent relates to increase the bioavailability of different nutritional supplements in Badmaev et al.
absorption enhancer combination with piperine (synthetic as well as extract) by increasing the cross over (1997)
through various biological barriers such as BBB, respiratory lining and likewise. The
invention reveals the mechanism of thermogenesis to increase the lean body mass.
3. US5616593A Compositions containing piperine The invention relates to the addition of piperine (as bioenhancer) in a fixed dose of 5 Patel et al.
mg/kg with several pharmaceutical compositions related to different diseases to achieve (1994)
the minimum possible dose to avoid the risk of toxicity as the result of clinical trial.
4. EP0935964A1 Pharmaceutical compositions containing The invention related to incorporate piperine and a selective COX-II and lipoxygenase Singh A et al.
NSAIDs and piperine (LOX) to increase the bio efficacy of the available dosage forms and/ or derivatives (1999)
containing the COX-II and LOX inhibitors. In addition, deals with the side effects related to
the dose of these agents.
5. IN2004MU00711A A novel method for the isolation of piperine The invention was principally relates to a novel method for the extraction and isolation of M Rajani et al.
from piper nigrum and piper longum piperine; the method deals with the enhanced purity and better yield. (2004)
6. US6365601B1 Process for extraction of piperine from piper The invention discloses a small and a rapid method for the extraction of piperine in two Raman G et al.
species simple steps using a hydrotropic solution as a vehicle. (2002)
7. EP2519511B1 Novel piperine derivatives as GABA-a The invention discloses a wide range of piperine derivatives as an effective treatment Hering S.
receptors modulators related to epilepsy, anxiety and insomnia by acting upon the GABAA receptor without (2015)
activating TRPV-1 receptor.
8. CA2735844C Use of piperine and analogues thereof in the The invention discloses the synthesis and identification of the novel derivatives of piperine Amla R et al.
prevention of skin cancer and the evaluation of these compounds in the treatment of vitiligo (melanocyte (1999)
proliferation).
9. EP0650728A1 Pharmaceutical compositions containing The invention discloses the effect of piperine on the bioavailability in combination with an Singh R K et al.
piperine and an anti-tuberculosis or Antitubercular and Anti-leprosy drug. The invention also relates to the reduction of dose on (1993)
anti-leprosy drug the effective cost of the treatment.
10. US2475980A Light-sensitive piperine compositions The invention relates to the conversion of piperine into a ‘piperite’ a resinous material Murray
obtained by the reaction of piperine synthesis with the exposure of light and heat or both. Alexander
Further, this material could be used in the photochemical reaction in the presence of an et al. (1949)
accelerator.

10
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Fig. 6. Most active compounds of piperic acid derivatives as GABA-A receptor inhibitor.

Fig. 7. Piperine derivatives as radical scavenging agent using DPPH.

volunteers. One compartment model was used for the study. In this study, 20
human volunteers (14 men and 6 women) were selected and were treated con-
tinuously for three consecutive days with midazolam 10 mg/kg and piperine 15
mg/kg. Results show an effective increase in the half-life and decreased clear-
ance of piperine when compared to placebo [105].
4.6. Synthesis and biological activity of piperic acid derivatives
Schöffmann A. et al. [106] reported piperine as a potential γ-
aminobutyric acid type-A (GABA-A) receptor inhibitor. In the study, 76 dif-
ferent synthetic analogues of piperine were synthesized by substituting pi-
peridine ring with various amine derivatives and were tested against the
GABA-A receptor. The results of the study showed compound 15
([(2E,4E)-5-(1,3-benzodioxol-5-yl)-N, N-dipropyl-2,4-pentadienamide]) to
be a potent inducer of the GABA-A receptor modulation. On the contrary,
compound 16 ([(2E,4E)-5-(1,3-benzodioxol-5-yl)-N,N-dibutyl-2,4-
pentadienamide]) displayed highly potent GABA-A receptor modulation.
It has been observed that both the synthesized analogues were not able to
activate the transient receptor potential vanilloid type-1 receptor (TRPV-
1) and both the analogues are responsible for inducing the pronounced
tranquilizer activity with a small amount of sedation [106].
(Fig. 6.)
Takao K. et al. reported the synthetic derivatives of piperine as the free
radical scavenging agent using 1,1-diphenyl-2-picrylhydrazyl (DPPH). The
activity of these amide derivatives acted against α-glucosidase inhibitors
and were found to be active when tested for their DPPH free radical scav-
enging and α-glucosidase inhibition. Compound 17, 18, and 19 were
found to be potent free radical scavenging agents; compound 21 and 22
were found to be active as the α- glucosidase inhibitors. The hydrophobic-
ity of the conjugated amines was the important determinant factor for α-
glucosidase inhibition. On the other hand, compound 18 (EC50: 28 μM
and IC50: 46 μM) and compound 19 (EC50: 20 μM and IC50: 46 μM) had
both α-glucosidase and DPPH free radical scavenging activity. Compound
20 was not found to be active against DPPH free radical activity [107].

Fig. 8. Piperine derivatives as insecticidal activity.

11
A. Tiwari et al.
Fig. 9. 1, 2, 4 triazole-3-thione piperamide derivatives of piperine against
Trypanosoma cruzi.
(Fig. 7.)
Paula V. F. et al. reported that piperine and its synthetic derivatives
show insecticidal activity. A series of 16 synthetic compounds were pre-
pared and were tested against Ascia monuste orseis, Acanthoscelides obtectus,
Brevicoryne brassicae L, Cornitermes cumulan and Protopolybia exigua. The re-
sults showed that six synthetic compounds were active against A. monuste
orseis species of the insects. Compound 23 (mortality: 97.5%) and com-
pound 24 (mortality: 95%) were found to be extremely active. Compound
25, 26, and Control had the mortality rate between 40.0% and 45.0%.
Compound 1, 26, 27, and 28 were tested against 4 species of insects with
the mortality rate between 20% and 23% [108].
(Fig. 8.)
Franklim T. N. et al. reported the activity of 1, 2, 4 triazole-3-thione
piperamide derivatives having potent trypanosomal activity against prolifera-
tive forms of Trypanosoma cruzi. Compound 29 showed potent activity [109].
(Fig. 9.)
Ribeiro et al. made structural modifications in the basic structure of piper-
ine. These compounds were tested against Trypanosoma cruzi. Results showed
that compound 30 was less toxic, which indicates the importance of the ali-
phatic side chain extension. In compound 31, removal of conjugated double
bonds showed no significant effect on trypanosomal activity, the substitution
of piperidine ring with diisopropyl (compound 32), and morpholyl group
(compound 33) resulted in a loss of activity on epimastigotes. Finally, the
Fig. 10. Structural modifications on piperine against Trypanosoma cruzi.
Fig. 11. Dipeptidyl boronic acid derivatives of piperine as antimicrobial agent.
12
Medicine in Drug Discovery 7 (2020) 100027
reduction of the carbonyl amide group, which gave allylic amine (compound
34), retained the significant toxicity against parasites [110].
(Fig. 10.)
Venugopal D. V. et al. reported newly synthesized derivatives of piper-
ine of dipeptidyl boronic acid as antimicrobial and anticancer agents.
These synthesized compounds were tested against HeLa cervical cancer,
MCF-7 breast, and MIA PaCa-2 pancreatic cancer cell lines. Antimicrobial
activity of piperine analogues was tested against seven bacterial and five se-
lected fungal species. The results suggest that compound 35 was more po-
tent in the anticancer screening of the compounds. On the other hand,
compound 36 was found to be active and more potent against the antimi-
crobial species Aspergillus Fumigates at the inhibitory concentration of 62
μg/ml [111].
(Fig. 11.)
Sangwan P.L. et al. reported structural modifications on piperine and its
synthetic analogues as potent Nor-Alpha efflux pump inhibitors. The struc-
tural modification at the C-4 position, by alkyl chain as well as the 1,3-
benzodioxol ring resulted in 4-fold decrease in the minimum inhibitory con-
centration (MIC) of ciprofloxacin activity against staphylococcus aureus, while
piperine shows significant reduction in the activity by two folds compared
with piperine derivatives (Compound 37, 38, 39, 40, 41, 42, and 43) [112].
(Fig. 12.)
Faas L. et al. presented a report suggesting that piperine can be a potent
therapeutic agent in the treatment of vitiligo. Piperine and two of its syn-
thetic derivatives (compound 44 and 45) tested on animal models were
compared with the conventional vitiligo therapies. Results suggested that
the combination of piperine and its synthetic derivatives significantly in-
creased the response to pigmentation with the potential reduction of skin
cancer [104].
(Fig. 13.)
4.7. Piperine derivatives as an antineoplastic agent
Ramesh A. et al. presented evidence of piperine and its synthetic ana-
logues as anticancer and antimicrobial agents. In the study, five analogues
of 5-(3,4- methylenedioxyphenyl)-2,4-pentadienoic acid were prepared
and tested against cancer cell lines (Hep-G2 and MCF-7) by conducting
A. Tiwari et al.
Fig. 12. Piperine as Nor-Alfa efflux pump inhibitor.
Fig. 13. Piperine and its synthetic derivatives as anti-vitiligo agents.
the MTT assay using ELISA reader. Compound 46 was active with an IC50
value of 1.25 μM against breast cancer cell line (MCF-7) whereas, com-
pound 47 had IC50 value of 0.89 μM against liver cancer cell line (Hep-
G2). On the contrary, all 5 compounds tested for the antimicrobial activity
by the Disc Diffusion Method against five microbial species namely Bacillus
subtilis, Enterococcus faecalis, Escherichia coli, Vibrio cholerae, and Pseudomo-
nas aeruginosa against ciprofloxacin as the reference compound showed
that these synthesized piperic acid amides were potent against these micro-
bial species [113].
(Fig. 14.)
Sakagami H et al. reported some of the piperic acid esters and their
quantitative structure cytotoxicity activity against four different human
oral squamous cell carcinoma (OSCC) followed by the cytotoxicity assay.
In the study, compound 48 was found to be active and was selected as
the lead compound for the synthesis of more potent analogues against
OSCC [114].
(Fig. 15.)
Samykutty A. et al. reported the effect of piperine and its mechanism to
act as a potential inhibitor of prostate cancer in a dose-dependent manner.
The proliferation of prostate cancer cells (LNCap), PC-3, 22RV1, and DU-
145 followed by different types of assays, namely cell line, cell viability
assay, caspase activation assay, apoptotic detection by staining with
annexin-V-FITC, western blot analysis, prostate-specific assay, and Boyden
Fig. 14. Most active piperine derivative as anticancer agent.
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Medicine in Drug Discovery 7 (2020) 100027
Fig. 15. PIperic acid ester in the treatment of OSCC.
chamber assay was also reported. Results of these studies suggest that
annexin-V-staining showed that the treatment of prostate cancer cells
with piperine could eventually induce apoptosis. Piperine was found to
be a responsible factor in disrupting the expression of the androgen recep-
tor. Lastly, Xenograft model using nude mice was developed using prostate
cancer cells and treated with piperine [115], (Table. 5).
4.8. Piperine derivatives as anti-tubercular agent
Hegeto L. A. et al. showed the antitubercular effect of piperine on the
overexpression of the mycobacterial putative efflux protein (Rv1258c).
On the other hand, the 3D structure and binding affinity of Rv1258c were
analyzed using In silico molecular modeling technique. In the study, the
MIC for the anti-tubercular agents (rifampicin, ethambutol, and streptomy-
cin) was evaluated, alone and in combination with piperine using Resazurin
as a reference followed by the microtiter assay. The cytotoxicity assay was
carried out on VERO cells and J774 A1 microphages. The synergism in ef-
flux pump inhibitor (EPI) activity was observed when combined with piper-
ine (RIF + PIP and SM + PIP) [116]. Philipova I et al. reported the activity
of some of the piperine analogues against mycobacterium tuberculosis
(H37Rv). The results suggest that all nine synthetic compounds of piperine
tested against H37Rv displayed better antitubercular activity than etham-
butol. One of the selected piperine amide derivatives showed more activity
than isoniazid. Compounds 48 and 49 had an IC50 value of 0.17 μM and
0.18 μM, respectively. Compound 48 (most potent analogue of piperine)
displayed a low cytotoxic effect on the human embryonal renal cells.
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Table 5
Other Therapeutic Effects of piperine
Sr. Biological Target In -vitro Tests Conducted In- Vivo Experimental Model Inference of the study Reference
No. Activity

1. Anticancer Cervical Anti-proliferative (HeLa), Cell Viability, - Piperine significantly inhibits the cell [131]
Adenocarcinoma Reactive oxygen species (ROS), Mitochondrial proliferation by reduction in the viability of
Membrane potential (MMP), DNA cells, ROS Increment, inducing cell shrinkage,
Fragmentation, Wound Healing, Cell cycle DNA fragmentation also by blocking the cell
Arrest, Caspase – 3 activation, Nuclear cycle and activating the Caspase-3 activity
Apoptosis.
Breast MTT cell viability assay (EMT6/P cells), Mouse Syngraft Piperine in combination with Thymoquinone [132]
Carcinoma VEGFR expression, DeadEnd TUNEL (EMT6/P cells) can be show synergistic effects for the
(piperine In Colorimetric Assay, Caspase- 3 induction. Histopathological, Biochemical inhibition of breast cancer by inducing the
combination tests. apoptosis and inhibiting the angiogenesis.
with
Thymoquinone)
Human Oral MTT cell viability assay (KB cells), ROS, DAPI - Piperine act by triggering the apoptosis [133]
Squamous analysis, MMP, cell cycle arrest, caspase- 3 through the cell cycle arrest and inhibiting
Carcinoma activation, modulation of cellular DNA the mitochondrial oxidative stress.
content, induction of G2/M phase arrest.
Prostate cancer LNCaP and PC-3 cells, Electrophysiology, - Piperine exhibits multivariate effects on the [134]
Cytotoxicity, Cell viability, Cell cycle, prostate cancer cell lines in dose dependent
analysis, analysis of piperine on the voltage manner.
gated K+ current modulation, apoptosis
Hepatocellular MTT assay (Hep G2 cells), antioxidant Chemical Induced Piperine exhibits the pro-oxidant mediated [135]
Carcinoma analysis, apoptosis, Catalase analysis (CAT) (Diethylnitrosamine) apoptosis by inhibiting the central molecule
analysis, reactice oxygen species analysis, hepatocellular carcinoma, tumor of defense system.
MMP assay, Western Blot (procaspase- 3, marker analysis, biochemical
Cleaved Caspase- 3, Cleaved Caspase-9, β- and histopathological analysis.
actin)
Triple Negative MTT Assay (MDA- MB- 231 cell line), Tumour Xenograft Model Piperine inhibits the growth and motility rate [136]
Breast Cancer spheroid culture and acid phosphate assay, (MDA-MB-231 Cells) of TNBC carcinogenic cells
(TNBC) cell proliferation using flow cytometry, cell
cycle analysis, apoptosis assay, western blot
analysis, Wound healing assay, Quantitative
real time PCR
Breast Cancer MTT assay (MCF-7, MDA-MB-231 Cell lines), - Piperine effectively inhibits the cell [137]
Caspase-3 activation, wound healing assay, proliferation and migration of cells , induces
Transwell migration assay, Quantitative real the apoptosis process in the HER-2 gene
time PCR, Western Blot analysis, Trasfection expressing breast cancer cells, reduction in
and Luciferase assay, the Epithilial Growth Factor (EGF) induces
MMP-9 expression.
Human Cell viability and Cytotoxicity Test (A375SM, Tumor Xenograft Animal Model Piperine inhibits the growth of the A375SM, [138]
Melanoma Cells A375P Cell lines), DAPI Analysis, Western (A375SM, A375P Cell lines) A375P Cells at variable concentrations due to
Blot analysis, Tunel Test, the increased apoptosis by inhibiting the
melanoma cancer cells
2. Antitubercular Mycobacterium Cell proliferation assay; cytokine estimation - Piperine upregulates the Th1 immunity and [139]
Activity tuberculosis by ELISA; MØ nitrite production—Griess. exhibits synergistic effects by combining with
H37Rv Protective efficacy of PIP in mice infection: rifampicin in immunocompromised patients
FC; VCFUC; RT-qPCR of tuberculosis.
Mycobacterium MIC (Checkboard Assay), Time kill Curve - The combination of RIF + PIP and SM+ PIP [116]
tuberculosis assay, Accumulation of ethidium bromide, shows synergistic effects on efflux pump
H37Rv Cytotoxicity assay inhibition (EPI) activity but less effective in
killing of the M. tuberculosis H37Rv.
Mycobacterium MIC determination, Modulation assay, EtBr - Piperine increases the antibacterial effect of [140]
smegmatis accumulation assay by the fluorometric rifampicin in time kill studies and enhances
method, EtBr efflux assay by the fluorometric the post antibiotic effect.
method
Mycobacterium Rifampicin/piperine combination studies, - Piperine plays a key role in inhibiting the [118]
tuberculosis Time–kill studies, Selection of resistant Rv1258c (MDR TB) by lowering the dosage
Rv1258c mutants, Post-antibiotic effect (PAE), frequency, increase in the mutant frequencies
(Multidrug Selection and susceptibility of resistant and by enhancing the rifampicin efficacy
resistant M. mutants, qRT–PCR analysis of the putative against M. tuberculosis.
tuberculosis) efflux pump protein
M. tuberculosis MABA - Chabamide a dimer of piperine shows the [141]
(H37Ra strain) antimalarial activity against plasmodium
falciparum and Antitubercular activity against
M. tuberculosis
M. tuberculosis MABA; checkerboard - The bioassay study of piperine revealed a [142]
Resistant to novel strategy of efflux pump inhibition and
ofloxacin synergistic effects of piperine in combination
with C. roseus

Quantitative structure-activity relationship revealed that the replacement Sharma et al. reported the efficacy of piperine as a potent inhibitor of
of piperidine ring with quaternary ammonium moiety is the primary and putative multidrug-resistant efflux pump (Rv1258c) of Mycobacterium tu-
concomitant requirement for the antitubercular effect [117]. berculosis effectively enhanced the antibacterial activity of rifampicin and
(Fig. 16.) increased the postantibiotic effect of rifampicin. Moreover, piperine

14
A. Tiwari et al.
Fig. 16. Synthetic analogues of piperine as anti-tubercular agent.
showed a three- to sixfold increase overexpression of multidrug-resistant
putative (EPI) activity [118]. Linn et al. reported piperine as a repigmenting
agent specifically for the treatment of vitiligo against cultured melanocytes.
Aqueous extract of piperine (0.1 mg mL-1) was administered in mouse me-
lanocytes. Results revealed that the administration of aqueous extract
shows significant stimulation of growth in these melanocytes (300%).
Growth augmentation using piperine was effectively repressed using RO-
31-8220 selective protein tyrosine kinase C inhibitor [119].
Faas L. reported that piperine can be used as a potential treatment ther-
apy for vitiligo. In the presented work, a mice model was used to demon-
strate the effect of piperine and two of its synthetic derivatives (5-(3,4-
methylenedioxyphenyl)-2,4-pentadienyl cyclohexylamine ({rCHP}) on vit-
iligo by topical application in combination with ultraviolet radiation in
lower doses led to a significant increase in pigmentation [120].
5. Formulation Technologies and enhancement of therapeutic
impacts
Recent advancements in nanotechnology have proven that nanoparticles
acquire great potential as drug carriers. Nanotechnology is a rapidly growing
area of importance and interest, incorporating a wide range of research fields.
Nanoscience primarily deals with synthesis, characterization, exploration,
and the exploitation of nanostructured materials. These materials were char-
acterized by at least one dimension in the nanometer range. The processing,
structure, and properties of materials with grain size in tens to several hun-
dreds of nanometer range are research areas of considerable interest over
the years. Nanotechnology is also efficient in improving the bioavailability
of drugs, protect therapeutic agents from physiological barriers, and enable
the development of new classes of bioactive macromolecules.
Elnaggar Y. S. et al. reported piperine loaded chitosan nanoparticles in
the treatment of Alzheimer’s disease. In this study, dose reduction ulti-
mately led to fewer side effects. Masking of the pungent attribute of piper-
ine was successfully achieved by encapsulation into nanoparticles [121].
Pachauri M. et al. reported piperine-loaded PEG-PLGA nanoparticles in
the targeted therapy of drug-resistant breast cancer as the anti-
proliferative action in conjugation with aptamer mucin. The APT-P-PEG-
PNP nanoparticles have shown remarkable anti-proliferative action against
breast cancer cell line MCF-7. Results of this study suggested that APT-P-
PEG-PNP nanoparticles could be used as a potential therapeutic agent in
the combined drug therapy [122]. Bhalekar M. R. et al. reported the use
of solid lipid nanoparticles (SLN) of piperine in the treatment of rheumatoid
arthritis using the melt emulsification method. The development of SLN
was successfully prepared, and the formulated SLNs were given orally as
well as topically to animals. The obtained results showed significant activ-
ity in disease-modifying rheumatoid arthritis drugs by using Complete
Freund’s Adjuvant as an inducing agent by both oral as well as by topical
routes [123]. Shaikh et al. reported that the nanoparticle encapsulation im-
proved oral bioavailability of curcumin by at least ninefold when compared
to curcumin administered with piperine as the absorption enhancer [124].
C. Moorthi et al. reported several methods for the preparation and char-
acterization of curcumin-piperine dual drug-loaded nanoparticles [125].
Jain et al. reported the preparation of biodegradable polymeric controlled
release nanofibrous patch using poly-(ε-caprolactone) (PCL) and gelatin
(GEL) blended using the electrospinning method for the treatment of can-
cer. The characterization of the nanofibrous patch of piperine was elabo-
rated using different spectroscopic methods, namely SEM and FTIR, and
15
Medicine in Drug Discovery 7 (2020) 100027
particle size. Later the in vitro release kinetics of the prepared formulation
was carried out, showing sustained release of the piperine-loaded PCL/
GEL patch releases 50% of piperine in 72 h. Furthermore, anticancer activ-
ity was observed by examining reduction in the cell viability and growth of
HeLa (cervical cancer) and MCF-7 (breast cancer) cell lines [126]. Elnaggar
Y. S. et al. reported the preparation of T-Cubes loaded with piperine and
tween for the treatment and toxicological evaluation on Alzheimer’s dis-
ease. In the research work, the bioactive nanocarriers targeting the brain
were prepared using different bioactive surfactants, namely tween-80,
poloxamer, and carriophor. The characterization of the prepared piperine
cubosomes was performed by particle size, Zeta potential, polydispersity
index, in vitro release studies, and entrapment efficiency. Later, the pharma-
cological evaluation of the prepared cubosomes was examined. Evaluation
of the toxicological parameters in the brain using the assessment of apopto-
sis and tumor necrosis factor-α inflammation. Hepatotoxicity and renal tox-
icological studies were carried out simultaneously [127].
6. Role of Pepper Species in Traditional Medicine
As a traditional medicine, black pepper and long pepper (Genus: piper;
Family: Piperaceae) are widely used in various systems of traditional med-
icine. In Ayurveda, black pepper is used in the treatment of cough, cold, and
sore throat. While in other systems of medicines, the peppercorns are used
for the treatment of indigestion, plethora, insomnia, and diseases related to
the heart. Vladimir Badmaev and Muhammed Majeed et al., (US 5,972,382;
1999) reported some of the traditional uses of pepper species. “Trikatu” an
Ayurvedic formulation containing Black pepper (piper nigrum), Long pepper
(piper longum) and Ginger (Zingiber officinale) is used individually or collec-
tively to enhance the bioavailability of a large number of nutrients includ-
ing ingredients of vasaka leaves, vasicine, sulfadiazine on both animal
and human volunteers. The study reveals that piperine an active constituent
of black pepper and long pepper, plays a significant role in increasing the
bioavailability of the drug. Furthermore, the effect of piperine on infertility
was investigated using laboratory animals [143].
Mechanism of Action: Black pepper and long pepper are widely used
in the traditional (Ayurveda, Siddha, Unani, and Tibetan) system of medi-
cine; and piperine, an active alkaloid present in pepper species is used as
a bioenhancer. The bioenhancing effect of pepper species in traditional as
well as modern medicine may be due to two major mechanisms; first, be-
cause of a nonspecific mechanism in which the rapid absorption of the
drug may be observed by the decreased secretion of HCl. It also enhances
the effect of gamma-glutamyl transpeptidase and by increasing the blood
supply to the GI tract [144]. Secondly, by inhibiting the enzymes involved
in biotransformation drugs and decreasing the rate of elimination. Atal C. K.
et al. reported the in vitro and in vivo interaction of piperine with the drug
metabolizing enzymes present in hepatic tissues. The study revealed that,
piperine was found as an inhibitor of aryl hydrocarbon hydroxylation
(AHH), ethylmorphine- N- demethylation, and 7- ethoxycoumarin-O-
deethylation in rat postmitochondrial supernatant in a dose-dependent
manner [145].
6.1. Ayurveda
In Ayurveda, the concept of bioavailability enhancers/biopotentiators is
called “yogavahi.” Bose first reported the concept of bioenhancers in 1929;
he reported the enhancement in the antiasthmatic activity of vasaka
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Table 6
Ayurvedic Formulations containing piperine
Sr. Formulation Composition Therapeutic Use Reference
No.

1. Trikatu Churna Black Pepper (piper nigrum Linn.), Long pepper (piperi longum Linn.), Traditionally used to for the restoration of Kapha, Vata, and Pitta. In [146–147]
(Major Ginger (Zingiber officinale Rosc.) various other types of fevers, disorders related to abdomen and GI tract,
Decoction) pyrexia, dysentery, bronchitis, asthma and sleep disorders. As a
bioavailability enhancer for (Vasicine, Sparteine)
2. Pippalyëdyësava Long pepper (Piper longum), Black pepper (Piper nigrum), Piper chaba malabsorption syndrome, abdominal lump, emaciation, pthisis, piles, [148]
urticarial, anaemia
3. Jawa rishai Black pepper (P. nigrum), Ginger (Zingiber officinale), Embilia ribes, Mint Stimulant, Carminative, Expectorant, genito urinary diseases, diuretic, [149]
Thurushi (Menthe sativa), and lemon juice Gonorrhea
4. Kanakasava Pippali (Long Pepper) – Piper Longum, Sonth (Ginger Rhizome) – Bronchodilator, Expectorant, Analgesic, Antispasmodic, Sedative, [150]
Zingiber Officinale, Mulethi – Glycyrrhiza Glabra, Vasaka – Adhatoda Cardiac stimulant, Soporific (Sleep inducing), Digestive
Vasica, Shuddha Datura Panchang – Purified Datura Metal (All Parts)
5. Panchakola Pippali (Piper longum), Pippalimoola (Piper longum), Chavya (Piper Antipyretic, analgesic, anti-inflammatory, appetizer, digestive, and [151]
Churna retrofractum), Chitraka (Plumbago zeylanica), Shunti (Zingiber officinale) carminative.
Vardhamana Pippali (Piper longum), Honey, ghee (Clarified Butter) Bronchitis, pthisis, asthma, piles, anemia, chronic rhinitis, intermittent [152]
Pippali Rasayana fever, splenic disorders, obstinate abdominal diseases, rheumatoid
arthritis
7. Avipattikara Black pepper (Piper nigrum), Ginger (Zingiber officinale), Amla (Emblica Diarrhea, Gastritis, Heartburn, Indigestion, Ulcers, Peptic Ulcers. [153–154]
Churna officinalis), Behada (Terminalia belerica), Cardamom (Elettaria
cardamomum), Clove (Syzygiumaromaticu)
8. Decoction Piper guineense Linn. Fungal infections, asthma, antipsychotic, respiratory infections, [155–159]
(seeds) infertility (female), bronchitis, syphilis
Decoction Piper guineense Anticonvulsant, sexually transmitted diseases (STDs), dysentery,
(Leaves) bronchitis.
9. Cold maceration Piper guineense Skin diseases, Mouth infection. [160]
(fruits and
leaves)
10. Talisapatradi Talisa (Abies webbiana), Maricha (Piper nigrum), Shunti (Zingiber Respiratory diseases, cough, cold, burning sensation in palms and feet, [158]
churnam officinale), Pippali (Piper longum), Vamshalochana (Bambusa bambos) low digestion power, loss of sensation in tongue, pain in abdomen,
flanks, anorexia, bleeding from nose

(Adhatoda vasica) when combined with Pippli (piper longum). The role of
yogavahi in Ayurveda is described as the agent used to enhance bioavail-
ability, tissue distribution, and efficacy of drugs, specifically with drugs
having poor bioavailability and decreasing adverse effects. In Ayurveda,
some special biopotentiators termed as Anupaan (combination of food
products with the medicament to increase the effect of the medication, e.
g., Amrit Dhara) and Sepaan (a vehicle that is used in the manufacture of
medication to increase the potential of the medicament, e.g., ghee, honey,
and panchgavya). There are number of ayurvedic preparations available
in Ayurveda that contain these yogavahis to increase the bioavailability
of poorly absorbed drugs when combined with principle medicament
(Table. 6).
Piperine is widely used as a bioavailability enhancer when combined
with various xenobiotic agents used in modern medicine to treat major dis-
eases. Johri R. K. and co-workers reported the medicinal properties of
“Trikatu” along with the pharmacological values of piperine.
6.2. Unani system of medicine
Filfil siyah (black pepper; piper nigrum), is an integral part of the unani
system of medicine, and is used to treat various different ailments (Table 7).
6.3. Siddha System of Medicine
Piper nigrum, an oldest spice and a major constituent in several siddha
drugs has a significant role in traditional Indian system of medicine. Classi-
cal siddha preparations that contain black pepper as a medicine are
amukkara choornam, Nilavembu kudineer, and Trikaduku choornam, hav-
ing pepper species as active constituents. The health benefits of pepper are
given in Table 8. (See Table 9.)
6.4. Marketed Formulation of piperine
Resorine [Fig. 17] is a marketed formulation (developed by RRL, Jammu
and marketed in India by Cadila Pharmaceuticals Ltd. Ahmedabad) containing
rifampicin (200 mg), isoniazid (300 mg), and piperine (10 mg). The medication
was approved for the treatment of pulmonary tuberculosis. It was found equiv-
alent with commercially available isoniazid tablets. The study reveals that the
dose of rifampicin was decreased by nearly half (from 450 mg to 200 mg) of
the primary dose of isoniazid formulation when combined with piperine.
Vora A et al. conducted a pilot study to determine the role of Resorine in
MDR pulmonary tuberculosis on human volunteers. In the study, 27 patients
with pulmonary tuberculosis were treated with a Resorine kit. The medica-
tion consisted of Resorine, ethambutol (800 mg), and pyrazinamide (750

Table. 7
Herbal Formulations containing piperine in Indian Pharmaceutical Market
Sr. Formulation Manufacturer Ingredients and Quantity Therapeutic Uses
No.

1. Pure Curcumin Simply Herbal Turmeric Powder (395 mg), Natural curcumin Anti-Inflammatory, Powerful antioxidant, prescribed as a
(Curcumin and Piperine Extract) extract (400 mg) and piperine (5 mg) dietary supplement.
2. Curcumin and Piperine Capsule Gaurav Marketing Curcumin and Piperine Anti-allergic, Natural detoxifier, Neuroprotective, Cholesterol,
Hepatoprotective, Gastric acid secretion
3. Premium Curcumin Pukhraj Weight Loss, Cancer, Protects Liver Health, Aids Digestion,
Eases Discomfort, inflammation and Prevents Arthritis
4. Curcumin 3 Health Aid Curcumin, Standardized Piperine (95%) Health supplement
5. Premium Biotin Capsules Simply Herbal Biotin (1000 mg), Bamboo extract (20 mg), Hair supplement
Piperine (5 mg)

16
A. Tiwari et al. Medicine in Drug Discovery 7 (2020) 100027

Table 8
Unani Formulations containing pepper
S. Formulation Composition Therapeutic Uses References
No.

1. Habb-e-Azaraqi Kuchla Biryan , Mirch Siyan (piper nigrum) Nervine tonic and stimulant. Specifically used in paralysis [160]
2. Jawarish Berg Suddab (Ruta graveolence leaf), Boora armani (Arminium bole), Filfil Siyah Stomachache, Colic pain. Improves digestion and removes [161]
Kamoni Sada (Piper nigrum fruit), Zanjabil (Zingiber officinalis rhizome), Zeera Siyah constipation.
Mudabbar (Carum carvi seed detoxified)
3. Habbe Kabid Naushader (Ammonium chloride) Namak Tuam (Sodium chloride) Namak Siyah Hepatitis, Dyspepsia, Constipation [162]
Naushadri (Sodium sulphate with Sodium chloride) Namak lahori (Sodium Chlorate) Haleela
Siyah (Terminelia chebula unripe fruit) Post Haleela Kabuli (Terminelia chebula
ripe fruit) Baobarang (Artemisia vulgaris seed) Filfil Siyah (Piper nigrum fruit)
Zanjabil (Zingiber officinalis rhizome) Suhaga (Borax) Zaranbad (Curcuma
zedoria root)
4. Jawarish Maghz-Tukhm-Kharpazah, Maghz-Tukhm-Kheyar, Tukhm-e-Karaf, Diuretic and Neuroprotective, Kidneys dysfunction, Weakness of [163]
Zarooni Sada Post-Beekh-e-Karafs, Post-Beekh-e-Karafs, Nankhah, Badiyan, Qaranfal, Filfi the digestive system, Liver dysfunction, Indigestion, Excessive
siyah, Aqarqarha, Darchini, Darchini, Zafran, Ood Hindi (Agar), Bisbasa urination and hydrospermia.
5. Habbe Gule Zanjabeel Zingiber officinalis, FilfilSiyah Pipernigrum Linn., GuleMadar Backache, Cervical, Spondylosis, Anti-inflammatory and analgesic [164]
Aakh Calotropis gigantea Linn., Barge Bans Bambusa arundinacea Willd. activity

Table 9
Siddha Formulations containing pepper
S. Formulation Composition Therapeutic Uses References
No.

1. Trikadagu Chukku (dried ginger), Milagu (pepper) and Thippili (long pepper) Analgesic and Anti-inflammatory properties. Digestive, carminative and [165]
Choornam used in treatment of Indigestion, gas trouble, constipation, bloating.
2. Kesari Lemon fruit, Milagu ( Piper nigrum), Daniya (C.sativum), Koshtam (Costus Vomitting, Giddiness, indigestion, Anorexia, Pitha diseases, Tastelessness, [166]
Lehyam speciosus), Chukku (Dried Ginger), Thaleesapatri (Taxus baccata), Tippili Flatulence.
(Piper longum), Jathikkai (Myrisica fragrans), Jathipathri (Mace), Ghee,
Sugar
3. Mezhugu Chukku (Zingiber officinale), Omam (Trachyspermum ammi L.), Manjal Painful conditions, fissures, leprosy, scrofula, rashes, alopecia, itches, hot [167]
(Curcuma longa L), Vaividangam (Embelia ribes Burm.f.), Vasambu (Acorus flush, glandular enlargements, cancer infections, piles, excessive pain in
calamus L.), Lavangam (Cinnamomum zeylanicum), Parangi pattai legs, slipped disc, back injury, skin diseases, psoriasis.
(Cucurbita pepo L.), Seran kottai (Semecarpus anarcadium L.), Kadukkai
thole (Terminalia chebula Retz.), Karum seeragam (Nigella sativa L.), Kaattu
seeragam (Centratherum anthelminticum L.), Siruthekku (Premna herbacea
Roxb.), Thaaleesapathiri (Taxus baccata), Thiraatchai (Vitis vinifera L.),
Thippili (Piper longum L.) Sitraraththai (Alpinia speciosa L.), Kottam
(Saussurea lappa Clarke) Valuluvai arisi (Celastrus paniculatus Willd),
Perum seeragam (Foeniculum vulgare Miller), Ela arisi (Elataria
cardamomum L.) Jadhikkai (Myristica fragrans Houtt.), Milagu (Piper
nigrum L.), Seeragam (Cuminum cyminum L.), Karboga arisi (Psoralea
corylifolia L.) Maasikkai (Quercus infectoria oliver) Thippili moolam (Piper
longum L.), Pirappan kizhangu ver (Calamus rotang L.), Ettikottai
(Strychnos nux –vomica L.) Thetraankottai (Strychnos potatorum L.)
Neermulli vidhai (Asteracantha longifolia L.Nees) Yellu (Sesamum indicum
L.) Kopparai thenkai (Cocos nucifera L.), Kollu (Dolichos biflorus L.) 34 Siru
chinni ver (Acalypha fruticosa Forsk.) Mutchangan ver (Azima tetracantha
Lam), Amukkara kizhangu (Withania somnifera L.), Aaakaasa garudan
kizhangu (Corallocarpus epigaeus Rotrl.), Kodiveli ver pattai (Plumbago
rosea L.), Kozhimuttai (Gallus domesticus L.), Panai vellam (Borassus
flabellifer L.)
4. Amukkara Ashwagandha (Withania Somnifera), Chukku (Zingiber Officinale), Thippili Nervine Tonic, Anti-inflammatory, Antitussive, Rasayana (Rejuvenating), [168,169]
choornam. (Long Pepper) (Piper Longum), Milagu (Black Pepper) (Piper Nigrum), Elam Aphrodisiac, Anti-depressant, Immunomodulatory, Antioxidant,
(Cardamom) (Elettaria Cardamomum), Sirunagapoo; Dalchini Anodyne, Cardioprotective, Mild Anti-asthmatic, Mild Anabolic effect
(Cinnamomum Verum), Kirambu Laung (Clove) (Syzygium Aromaticum),
Sakkarai (Saccharum Officinarum)

Fig. 17. Resorine: a marketed formulation of piperine.

17
A. Tiwari et al.
mg) per day. Medication was continued for two months. Later, patients re-
ceived a single tablet of Resorine for the next four months. The study con-
cluded that among 27 patients (smear-positive), 24 patients were found
sputum negative after treatment with Resorine in the first two months, re-
maining three patients out of which two patients were found to have mild
nausea and one patient had HIV-positive developed hepatitis [161,162].
Other than Ayurveda, there are numerous herbal formulations contain-
ing piperine in combination with other herbal extracts and are being used
in maintaining the lifestyle as a health supplement. Some of the herbal for-
mulations contain piperine in combination with other ingredients like
curcumin and keratin. Some of the formulations are given in Table. 8.
7. Conclusion and Future Prospect
Natural products have contributed to nearly half of all small molecules
approved as drugs in this decade. It has been suggested that the current
drug discovery approach of finding “new chemical entities” if shifted to
“combining existing agents,” may be helpful. Furthermore, higher dose,
poor absorption, low bioavailability, and poor patient compliance are the
major drawbacks of drugs. Black pepper consists of approximately 5-9%
piperine as an active chemical constituent. It was listed by FDA as a herb,
also recognized as safer agents for its use as a spice. Piperine can be given
at a dose of 15-20 mg/person/day in divided dose, which seems to be ex-
tremely less than the LD50 of the human dose. Piperine can be used to im-
prove the efficacy and reduce the dosing frequency of various xenobiotic
agents. By combining piperine with the medication, side effects and toxicity
of drugs can be reduced due to a lower dose and increased availability at the
site of action.
Along with the bioenhancing effect, piperine is well reported in Indian
medicine for the treatment of diseases such as rheumatoid arthritis, cancer,
asthma, inflammation, anxiety, depression, and infectious diseases. Piper-
ine significantly enhances the bioavailability of different drugs such as ri-
fampicin, simvastatin, ibuprofen, and omeprazole. In the current review,
isolation, and extraction, total synthesis, pharmacological activities of pip-
erine, and its derivatives and bioenhancing properties are discussed. The fu-
ture perspective is that piperine may be used along with poor ADMET drugs
to enhance the bioavailability, minimize the dose, and to possess synergistic
effects in the treatment of various disease states. Mechanism by which pip-
erine enhances the bioavailability may be a future goal. Furthermore, the
preparation of advance pharmaceutical formulations, targeted delivery,
and patient compliance may be the future goal in the area of bioenhancers.
The review may certainly provide insight details regarding work already
done concerning piperine. Both black pepper (Piper nigrum) and long pep-
per (Piper longum), important sources of piperine, is being used in large
amounts in traditional pharmacopeia even today. Despite long years of
use and the best experimental models and highly refined molecular and
cell biology research methods and tools available today, the mode of action
of piperine remains a mystery.
Acknowledgment
We owe our sincere gratitude to Dr. Shivajirao Kadam, Chancellor,
Bharati Vidyapeeth (Deemed to be University), Pune, for providing all nec-
essary facilities for the presentation of the work.
Declaration of interest statement
The authors declare that they do not have any conflicts of interest.
References
[1] Chopra B, Dhingra AK, Kapoor RP, Prasad DN. Piperine and its various physicochem-
ical and biological aspects: a review. Open Chem J. 2016;3. https://doi.org/10.
2174/1874842201603010075.
[2] Sozzi GO, Peter KV, Babu KN, Divakaran M. Capers and caperberries. Handb. herbs
spices. Elsevier; 2012; 193–224.
18
Medicine in Drug Discovery 7 (2020) 100027
[3] Pruthi JS. Quality assurance in spices and spice products, modern methods of analysis;
1999 ISBN: 9788170238966.
[4] Ravindran P. Black pepper: Piper nigrum. Boca Raton, Fla: CRC Press; 2003 ISBN
9789057024535.
[5] Hirasa K, Takemasa M. Spice science and technology. Boca Raton, Fla.: CRC Press;
1998 ISBN 9780824701444.
[6] Kozukue N, Park M-S, Choi S-H, Lee S-U, Ohnishi-Kameyama M, Levin CE, et al. Kinet-
ics of light-induced cis− trans isomerization of four piperines and their levels in
ground black peppers as determined by HPLC and LC/MS. J Agric Food Chem. 2007;
55:7131–9. https://doi.org/10.1021/jf070831p.
[7] Zachariah TJ, Parthasarathy VA. Black pepper. Chem spices, 196; 2008; 21 ISBN-13:
9781 845934057.
[8] Pruthi JS. Major spices of India: crop management and post-harvest technology. Major
Spices India Crop Manag Post-Harvest Technol. 1993. https://www.cabdirect.org/
cabdirect/abstract/20043186295.
[9] Correa EA, Högestätt ED, Sterner O, Echeverri F, Zygmunt PM. In vitro TRPV1 activity
of piperine derived amides. Bioorg Med Chem. 2010;18:3299–306. https://doi.org/
10.1016/j.bmc.2010.03.013.
[10] Hlavačková L, Janegová A, Uličná O, Janega P, Černá A, Babál P. Spice up the hyper-
tension diet-curcumin and piperine prevent remodeling of aorta in experimental L-
NAME induced hypertension. Nutr Metab (Lond). 2011;8:72. https://doi.org/10.
1186/1743-7075-8-72.
[12] Pradeep CR, Kuttan G. Effect of piperine on the inhibition of lung metastasis induced
B16F-10 melanoma cells in mice. Clin Exp Metastasis. 2002;19:703–8. https://doi.
org/10.1023/a:1021398601388.
[13] Li S, Lei Y, Jia Y, Li N, Wink M, Ma Y. Piperine, a piperidine alkaloid from Piper nigrum
re-sensitizes P-gp, MRP1 and BCRP dependent multidrug resistant cancer cells.
Phytomedicine. 2011;19:83–7. https://doi.org/10.1016/j.phymed.2011.06.031.
[14] Meghwal M, Goswami TK. Piper nigrum and piperine: an update. Phyther Res. 2013;
27:1121–30. https://doi.org/10.1002/ptr.4972.
[16] Zarai Z, Boujelbene E, Ben Salem N, Gargouri Y, Sayari A. Antioxidant and antimicro-
bial activities of various solvent extracts, piperine and piperic acid from Piper nigrum.
LWT-Food Sci Technol. 2013;50:634–41. https://doi.org/10.1016/j.lwt.2012.07.036.
[17] Tavares WS, Cruz I, Petacci F, Freitas SS, Serratilde JE, Zanuncio JC. Insecticide activ-
ity of piperine: Toxicity to eggs of Spodoptera frugiperda (Lepidoptera: Noctuidae) and
Diatraea saccharalis (Lepidoptera: Pyralidae) and phytotoxicity on several vegetables.
J Med Plants Res. 2011;5:5301–6. https://doi.org/10.21276/sajb.
[18] Storz P. Reactive oxygen species in tumor progression. Front Biosci. 2005;10:1881–96.
https://doi.org/10.2741/1667.
[19] Su HCF. Insecticidal properties of black pepper to rice weevils and cowpea weevils. J
Econ Entomol. 1977;70:18–21. https://doi.org/10.1093/jee/70.1.18.
[20] Miyakado M, Nakayama I, Yoshioka H, Nakatani N. The Piperaceae amides I: Structure
of pipercide, a new insecticidal amide from Piper nigrum L. Agric Biol Chem. 1979;43:
1609–11. https://doi.org/10.1080/00021369.1979.10863675.
[21] Bang JS, Choi HM, Sur B-J, Lim S-J, Kim JY, Yang H-I, et al. Anti-inflammatory and an-
tiarthritic effects of piperine in human interleukin 1β-stimulated fibroblast-like
synoviocytes and in rat arthritis models. Arthritis Res Ther. 2009;11:R49. https://
doi.org/10.1186/ar2662 Epub 2009 Mar 30.
[22] Mujumdar AM, Dhuley JN, Deshmukh VK, Raman PH, Naik SR. Anti-inflammatory ac-
tivity of piperine. Japanese J Med Sci Biol. 1990;43:95–100. https://doi.org/10.1186/
ar2662.
[23] Ghoshal S, Prasad BNK, Lakshmi V. Antiamoebic activity of Piper longum fruits against
Entamoeba histolytica in vitro and in vivo. J Ethnopharmacol. 1996;50:167–70.
https://doi.org/10.1016/0378-8741(96)01382-7.
[24] Mehmood MH, Gilani AH. Pharmacological basis for the medicinal use of black pepper
and piperine in gastrointestinal disorders. J Med Food. 2010;13:1086–96. https://doi.
org/10.1089/jmf.2010.1065.
[25] Lee SA, Hong SS, Han XH, Hwang JS, Oh GJ, Lee KS, et al. Piperine from the fruits of
Piper longum with inhibitory effect on monoamine oxidase and antidepressant-like ac-
tivity. Chem Pharm Bull. 2005;53:832–5. https://doi.org/10.1248/cpb.53.832.
[26] Bano G, Amla V, Raina RK, Zutshi U, Chopra CL. The effect of piperine on pharmaco-
kinetics of phenytoin in healthy volunteers. Planta Med. 1987;53:568–9. https://doi.
org/10.1248/cpb.53.832.
[27] Khatri S, Ahmed FJ, Rai P. Formulation and evaluation of floating gastroretentive cap-
sules of acyclovir with piperine as a bioenhancer. Pharma Innov. 2015;3:78.
[28] Khatri S, Awasthi R. Piperine containing floating microspheres: an approach for drug
targeting to the upper gastrointestinal tract. Drug Deliv Transl Res. 2016;6:299–307.
[29] Prasad R, Singh A, Gupta N, Tarke C. Role of bioenhancers in tuberculosis. Int J Heal
Sci. 2016;6.
[30] Park U-H, Jeong H-S, Jo E-Y, Park T, Yoon SK, Kim E-J, et al. Piperine, a component of black
pepper, inhibits adipogenesis by antagonizing PPARγ activity in 3T3-L1 cells. J Agric Food
Chem. 2012;60:3853–60. https://doi.org/10.1021/jf204514a Epub 2012 Apr 6.
[31] Ahmad N, Fazal H, Abbasi BH, Farooq S, Ali M, Khan MA. Biological role of Piper
nigrum L.(Black pepper): A review. Asian Pac J Trop Biomed. 2012;2:S1945–53.
https://doi.org/10.1016/S2221-1691(12)60524-3.
[32] Meghwal M, Goswami TK. Chemical composition, nutritional, medicinal and func-
tional properties of black pepper: A review. Open Access Sci Rep. 2012;1:1–5.
[33] Pal Singh I, Choudhary A. Piperine and derivatives: Trends in structure-activity rela-
tionships. Curr Top Med Chem. 2015;15:1722–34. https://doi.org/10.2174/
1568026615666150427123213.
[34] Dudhatra GB, Mody SK, Awale MM, Patel HB, Modi CM, Kumar A, et al. A comprehen-
sive review on pharmacotherapeutics of herbal bioenhancers. Sci World J. 2012;2012.
https://doi.org/10.1100/2012/637953.
[35] Cao X, Ye X, Lu Y, Yu Y, Mo W. Ionic liquid-based ultrasonic-assisted extraction of pip-
erine from white pepper. Anal Chim Acta. 2009;640:47–51. https://doi.org/10.1016/
j.aca.2009.03.029.
A. Tiwari et al.
[36] Sun X, Jin Z, Yang L, Hao J, Zu Y, Wang W, et al. Ultrasonic-assisted extraction of
procyanidins using ionic liquid solution from Larix gmelinii bark. J Chem. 2012;
2013. https://doi.org/10.1155/2013/541037.
[37] Shingate PN, Dongre PP, Kannur DM. New method development for extraction and iso-
lation of piperine from black pepper. Int J Pharm Sci Res. 2013;4:3165–70. https://
doi.org/10.13040/IJPSR.0975-8232.4(8).3165-70.
[38] Kanaki N, Dave M, Padh H, Rajani M. A rapid method for isolation of piperine from the
fruits of Piper nigrum Linn. J Nat Med. 2008;62:281–3. https://doi.org/10.1007/
s11418-008-0234-3.
[39] Raman G, Gaikar VG. Microwave-assisted extraction of piperine from Piper nigrum.
Ind Eng Chem Res. 2002;41:2521–8. https://doi.org/10.1021/ie010359b.
[40] Subramanian R, Subbramaniyan P, Ameen JN, Raj V. Double bypasses soxhlet appara-
tus for extraction of piperine from Piper nigrum. Arab J Chem. 2016;9:S537–40.
https://doi.org/10.1016/j.arabjc.2011.06.022.
[41] Hamrapurkar PD, Jadhav K, Zine S. Quantitative estimation of piperine in Piper
nigrum and Piper longum using high performance thin layer chromatography. J
Appl Pharm Sci. 2011;1:117–20.
[42] Kolhe SR, Borole P, Patel U. Extraction and evaluation of piperine from Piper nigrum
Linn. Inter J Appl Biol Pharma Tech. 2011;2:144–9.
[43] Sreevidya N, Mehrotra S. Spectrophotometric method for estimation of alkaloids pre-
cipitable with Dragendorff’s reagent in plant materials. J AOAC Int. 2003;86:1124–7.
[45] Mengal P, Mompon B. Method and plant for solvent-free microwave extraction of nat-
ural products; 2006. https://doi.org/10.1016/j.trac.2013.02.007.
[46] Gaikar VG, Raman G. Microwave-assisted extraction of piperine from Piper nigrum.
Ind Eng Chem Res. 2002;41:2521–8.
[47] Gigliarelli G, Pagiotti R, Persia D, Marcotullio MC. Optimisation of a Naviglio-
assisted extraction followed by determination of piperine content in Piper longum
extracts. Nat Prod Res. 2017;31:214–7. https://doi.org/10.1080/14786419.
2016.1217209.
[48] Naviglio D. Naviglio’s principle and presentation of an innovative solid–liquid extrac-
tion technology: extractor Naviglio®. Anal Lett. 2003;36:1647–59. https://doi.org/
10.1081/AL-120021555.
[49] Han D, Row KH. Recent applications of ionic liquids in separation technology. Mole-
cules. 2010;15:2405–26. https://doi.org/10.3390/molecules15042405.
[50] Poole CF, Poole SK. Extraction of organic compounds with room temperature ionic liq-
uids. J Chromatogr A. 2010;1217:2268–86.
[51] Yang L, Sun X, Yang F, Zhao C, Zhang L, Zu Y. Application of ionic liquids in the micro-
wave-assisted extraction of proanthocyanidins from Larix gmelini bark. Int J Mol Sci.
2012;13:5163–78. https://doi.org/10.3390/ijms13045163.
[52] Olalere OA, Abdurahman NH, Bin Mohd Yunus R, Alara OR, Akbari S. Evaluation of
optimization parameters in microwave reflux extraction of piperine-oleoresin from
black pepper (Piper nigrum). Beni-Suef Uni J Bas Appl Sci. 2018 Dec 1;7(4):626–31.
https://doi.org/10.1186/s40543-017-0118-9.
[53] Rathod SS, Rathod VK. Extraction of piperine from Piper longum using ultrasound.
Ind Crops Prod. 2014;58:259–64. https://doi.org/10.1016/j.indcrop.2014.03.
040.
[54] Khaw K-Y, Parat M-O, Shaw PN, Falconer JR. Solvent supercritical fluid technologies
to extract bioactive compounds from natural sources: a review. Molecules. 2017;22:
1186. https://doi.org/10.3390/molecules22071186.
[55] Mukhopadhyay M. Natural extracts using supercritical carbon dioxide. CRC press;
2000 DOI: ISBN 9780849308192.
[56] Sovová H, Jez J, Bártlová M, St'astová J. Supercritical carbon dioxide extraction of 731
black pepper. J Supercri Fluids. 1995;8:295–301.
[57] Schofield K. Hetero-aromatic nitrogen compounds: pyrroles and pyridines. Springer;
2013. https://doi.org/10.1007/978-1-4899-5892-1.
[58] Tsuboi S, Takeda A. A new synthesis of piperine and isochavicine. Tetrahedron Lett.
1979;20:1043–4. https://doi.org/10.2174/1874842201603010075.
[59] Ladenburg A. Ber Deutsch Chem Ges. 1885;18:3100. https://doi.org/10.1016/0031-
9422(90)85102-L.
[60] Olsen RA, Spessard GO. A short, stereoselective synthesis of piperine and related pep-
per-derived alkaloids. J Agric Food Chem. 1981;29:942–4. https://doi.org/10.1021/
jf00107a013.
[61] Mandai T, Moriyama T, Tsujimoto K, Kawada M, Otera J. Highly stereoselective
synthesis of (2E, 4E)-dienamides and (2E, 4E)-dienoates via a double elimina-
tion reaction. Tetrahedron Lett. 1986;27:603–6. https://doi.org/10.1016/
S0040-4039(00)84052-6.
[62] Sloop JC. Microscale synthesis of the natural products carpanone and piperine. J Chem
Educ. 1995;72:A25. https://doi.org/10.1021/ed072pA25.
[63] Chandrasekhar S, Reddy MV, Reddy KS, Ramarao C. Addition of carbon nucleophiles
to aldehyde tosylhydrazones of aromatic and heteroaromatic-compounds: total synthe-
sis of piperine and its analogs. Tetrahedron Lett. 2000;41:2667–70.
[64] Schobert R, Siegfried S, Gordon GJ. Three-component synthesis of (E)-α, β-unsaturated
amides of the piperine family. J Chem Soc Perkin Trans. 2001;1:2393–7. https://doi.
org/10.1039/B105745F.
[65] Inatani R, Nakatani N, Fuwa H. Structure and Synthesis of New Phenolic Amides from
Piper nigrum L. Agric Biol Chem. 1981;45:667–73. https://doi.org/10.1080/
00021369.1981.10864574.
[66] Han L-C, Stanley PA, Wood PJ, Sharma P, Kuruppu AI, Bradshaw TD, et al. Horner–
Wadsworth–Emmons approach to piperlongumine analogues with potent anti-cancer
activity. Org Biomol Chem. 2016;14:7585–93. https://doi.org/10.1039/c6ob01160h.
[67] Aswar U, Shintre S, Chepurwar S, Aswar M. Antiallergic effect of piperine on ovalbu-
min-induced allergic rhinitis in mice. Pharm Biol. 2015;53:1358–66. https://doi.org/
10.3109/13880209.2014.982299.
[68] Kim J, Lee K-P, Lee D-W, Lim K. Piperine enhances carbohydrate/fat metabolism in
skeletal muscle during acute exercise in mice. Nutr Metab (Lond). 2017;14:43.
https://doi.org/10.1186/s12986-017-0194-2.
19
Medicine in Drug Discovery 7 (2020) 100027
[69] Dong Y, Huihui Z, Li C. Piperine inhibit inflammation, alveolar bone loss and collagen
fibers breakdown in a rat periodontitis model. J Periodontal Res. 2015;50:758–65.
https://doi.org/10.1111/jre.12262.
[70] Liu Y, Yadev VR, Aggarwal BB, Nair MG. Inhibitory effects of black pepper (Piper
nigrum) extracts and compounds on human tumor cell proliferation, cyclooxygenase
enzymes, lipid peroxidation and nuclear transcription factor-kappa-B. Nat Prod
Commun. 2010;5.
[71] Hu D, Wang Y, Chen Z, Ma Z, You Q, Zhang X, et al. The protective effect of piperine on
dextran sulfate sodium induced inflammatory bowel disease and its relation with
pregnane X receptor activation. J Ethnopharmacol. 2015;169:109–23. https://doi.
org/10.1016/j.jep.2015.04.006.
[72] Bang JS, Choi HM, Sur B-J, Lim S-J, Kim JY, Yang H-I, et al. Anti-inflammatory and an-
tiarthritic effects of piperine in human interleukin 1β-stimulated fibroblast-like
synoviocytes in rat arthritis models. Arthritis Res Ther. 2009;11:R49. https://doi.
org/10.1186/ar2662.
[73] Narasimhamurthy K. Lack of genotoxic effects of piperine,(the active principle of black
pepper) in albino mice. J Food Saf. 1990;11:39–48. https://doi.org/10.1111/j.1745-
4565.1990.tb00037.x.
[74] D’Hooge R, Pei YQ, Raes A, Lebrun P. Anticonvulsant activity of piperine on seizures
induced by excitatory amino acid receptor agonists. Arzneimittelforschung. 1996;46:
557–60.
[75] Ren T, Wang Q, Li C, Yang M, Zuo Z. Efficient brain uptake of piperine and its pharma-
cokinetics characterization after oral administration. Xenobiotica. 2018;48:1249–57.
https://doi.org/10.1080/00498254.2017.1405293.
[76] Yun YS, Noda S, Takahashi S, Takahashi Y, Inoue H. Piperine-like alkamides from
Piper nigrum induce BDNF promoter and promote neurite outgrowth in Neuro-2a
cells. J Nat Med. 2018;72:238–45. https://doi.org/10.1007/s11418-017-1140-3.
[77] Al-Baghdadi OB, Prater NI, Van der Schyf CJ, Geldenhuys WJ. Inhibition of mono-
amine oxidase by derivatives of piperine, an alkaloid from the pepper plant Piper
nigrum, for possible use in Parkinson’s disease. Bioorg Med Chem Lett. 2012;22:
7183–8. https://doi.org/10.1016/j.bmcl.2012.09.056.
[78] Mori A, Kabuto H, Pei Y-Q. Effects of piperine on convulsions and on brain serotonin
and catecholamine levels in E1 mice. Neurochem Res. 1985;10:1269–75. https://doi.
org/10.1007/bf00964845.
[79] Chonpathompikunlert P, Wattanathorn J, Muchimapura S. Piperine, the main alkaloid
of Thai black pepper, protects against neurodegeneration and cognitive impairment in
animal model of cognitive deficit like condition of Alzheimer’s disease. Food Chem
Toxicol. 2010;48:798–802. https://doi.org/10.1016/j.fct.2009.12.009.
[80] Essa MM, Vijayan RK, Castellano-Gonzalez G, Memon MA, Braidy N, Guillemin GJ.
Neuroprotective effect of natural products against Alzheimer’s disease. Neurochem
Res. 2012;37:1829–42. https://doi.org/10.1007/s11064-012-0799-9.
[81] Srinivasan K. Black pepper and its pungent principle-piperine: a review of diverse
physiological effects. Crit Rev Food Sci Nutr. 2007;47:735–48. https://doi.org/10.
1080/10408390601062054.
[82] Bhat BG, Chandrasekhara N. Studies on the metabolism of piperine: absorption, tissue
distribution and excretion of urinary conjugates in rats. Toxicology. 1986;40:83–92.
https://doi.org/10.1016/0300-483x(86)90048-x.
[83] Bhat BG, Chandrasekhara N. Metabolic disposition of piperine in the rat. Toxicology.
1987;44:99–106. https://doi.org/10.1016/0300-483X(87)90049-7.
[84] Bhat BG, Chandrasekhara N. Interaction of piperine with rat liver microsomes. Toxicol-
ogy. 1987;44:91–8. https://doi.org/10.1016/0300-483X(87)90048-5.
[85] Badmaev V, Majeed M, Prakash L. Piperine derived from black pepper increases the
plasma levels of coenzyme Q10 following oral supplementation. J Nutr Biochem.
2000;11:109–13. https://doi.org/10.1016/s0955-2863(99)00074-1.
[86] Bhardwaj RK, Glaeser H, Becquemont L, Klotz U, Gupta SK, Fromm MF. Piperine, a
major constituent of black pepper, inhibits human P-glycoprotein and CYP3A4. J
Pharmacol Exp Ther. 2002;302:645–50. https://doi.org/10.1124/jpet.102.034728.
[87] Khan IA, Mirza ZM, Kumar A, Verma V, Qazi GN. Piperine, a phytochemical potentia-
tor of ciprofloxacin against Staphylococcus aureus. Antimicrob Agents Chemother.
2006;50:810–2. https://doi.org/10.1128/AAC.50.2.810-812.2006.
[88] Brenwald NP, Gill MJ, Wise R. Prevalence of a Putative Efflux Mechanism among Flu-
oroquinolone-Resistant Clinical Isolates of Streptococcus pneumoniae. Antimicrob
Agents Chemother. 1998;42:2032–5.
[89] Farghaly AA, Abo-Zeid MAM. Evaluation of the antimutagenic effect of vitamin C against
DNA damage and cytotoxicity induced by trimethyltin in mice. Nat Sci. 2009;7:1e7.
[90] Omri N, Yahyaoui M, Banani R, Messaoudi S, Moussa F, Abderrabba M. Ab-initio HF and
density functional theory investigations on the synthesis mechanism, conformational sta-
bility, molecular structure and UV spectrum of N’-Formylkynurenine. J Theor Comput
Chem. 2016;15:1650006. https://doi.org/10.1142/S0219633616500061.
[91] Abo-Zeid MA, Farghaly AA. The anti-mutagenic activity of piperine against
mitomycine C induced sister chromatid exchanges and chromosomal aberrations in
mice. Nat Sci. 2009;7:72–8.
[92] Gökalp F, Apjes I. A study on piperine, active compound of black pepper. J Eng Sci.
2016:29. https://doi.org/10.21541/apjes.66230.
[93] Bajad S, Bedi KL, Singla AK, Johri RK. Piperine inhibits gastric emptying and gastroin-
testinal transit in rats and mice. Planta Med. 2001;67:176–9. https://doi.org/10.1055/
s-2001-11505.
[94] Bedada SK, Boga PK. The influence of piperine on the pharmacokinetics of
fexofenadine, a P-glycoprotein substrate, in healthy volunteers. Eur J Clin Pharmacol.
2017;73:343–9. https://doi.org/10.1007/s00228-016-2173-3.
[95] Athukuri BL, Neerati P. Enhanced oral bioavailability of domperidone with piperine in
male wistar rats: involvement of CYP3A1 and P-gp inhibition. J Pharm Pharm Sci.
2017;20:28–37. https://doi.org/10.18433/J3MK72.
[96] Singh A, Verma S, Al Zarari NMH, Singh AP, Fuloria NKK, Fuloria S. Effect of Piperine
on Pharmacokinetics of Rifampicin and Isoniazid: Development and Validation of High
Performance Liquid Chromatography Method. J Appl Pharm Sci. 2018;8:72–81.
A. Tiwari et al.
[97] Hiwale AR, Dhuley JN, Naik SR. Effect of co-administration of piperine on pharmaco-
kinetics of β-lactam antibiotics in rats; 2002.
[98] Pattanaik S, Hota D, Prabhakar S, Kharbanda P, Pandhi P. Pharmacokinetic interaction
of single dose of piperine with steady-state carbamazepine in epilepsy y patients.
Phyther Res An Int J Devoted to Pharmacol Toxicol Eval Nat Prod Deriv. 2009;23:
1281–6. https://doi.org/10.1002/ptr.1937.
[99] Pattanaik S, Hota D, Prabhakar S, Kharbanda P, Pandhi P. Effect of piperine on the
steady-state pharmacokinetics of phenytoin in patients with epilepsy. Phyther Res
An Int J Devoted to Pharmacol Toxicol Eval Nat Prod Deriv. 2006;20:683–6. https://
doi.org/10.1002/ptr.1937.
[100] Auti P, Choudhary A, Gabhe S, Mahadik K. Bioanalytical method development, valida-
tion and its application in Pharmacokinetic studies of Verapamil in the presence of Pip-
erine in rats. Int J Pharm Res. 2018;10:119.
[101] Auti P, Gabhe S, Mahadik K. Pharmacokinetic studies of secnidazole in the presence of pip-
erine and its synthetic derivative. DOI: . 10.13040/IJPSR.0975-823 2.10(3).1455-61
[102] Auti P, Gabhe S, Mahadik K. Bioanalytical method development and its application to
pharmacokinetics studies on Simvastatin in the presence of piperine and two of its syn-
thetic derivatives. Drug Dev Ind Pharm. 2019;45:664–8. https://doi.org/10.1080/
03639045.2019.1569034.
[103] Bano G, Amla V, Raina RK, Zutshi U, Chopra CL. The effect of piperine on pharmaco-
kinetics of phenytoin in healthy volunteers. Planta Med. 1987;53:568–9. https://doi.
org/10.1055/s-2006-962814.
[104] Bano G, Raina RK, Zutshi U, Bedi KL, Johri RK, Sharma SC. Effect of piperine on bio-
availability and pharmacokinetics of propranolol and theophylline in healthy volun-
teers. Eur J Clin Pharmacol. 1991;41:615–7. https://doi.org/10.1007/BF00314996.
[105] Rezaee MM, Kazemi S, Kazemi MT, Gharooee S, Yazdani E, Gharooee H, et al. The ef-
fect of piperine on midazolam plasma concentration in healthy volunteers, a research
on the CYP3A-involving metabolism. DARU J Pharm Sci. 2014;22:8. https://doi.org/
10.1007/BF00314996.
[106] Schöffmann A, Wimmer L, Goldmann D, Khom S, Hintersteiner J, Baburin I, et al. Ef-
ficient modulation of γ-aminobutyric acid type A receptors by piperine derivatives. J
Med Chem. 2014;57:5602–19. https://doi.org/10.1021/jm5002277.
[107] Takao K, Miyashiro T, Sugita Y. Synthesis and Biological Evaluation of Piperic Acid
Amides as Free Radical Scavengers and α-Glucosidase Inhibitors. Chem Pharm Bull.
2015;63:326–33. https://doi.org/10.1248/cpb.c14-00874.
[108] de Paula VF, de A Barbosa LC, Demuner AJ, Piló-Veloso D, Picanço MC. Synthesis and in-
secticidal activity of new amide derivatives of piperine. Pest Manag Sci. 2000;56:168–74.
https://doi.org/10.1002/(SICI)1526-4998(200002)56:2<168::AID-PS110>3.0.CO;2-H.
[109] Franklim T, Freire-de-Lima L, de Nazareth Sá Diniz J, Previato J, Castro R, Mendonça-
Previato L, et al. Design, synthesis and trypanocidal evaluation of novel 1, 2, 4-
triazoles-3-thiones derived from natural piperine. Molecules. 2013;18:6366–82.
https://doi.org/10.3390/molecules18066366.
[110] Ribeiro TS, Freire-de-Lima L, Previato JO, Mendonça-Previato L, Heise N, de Lima
MEF. Toxic effects of natural piperine and its derivatives on epimastigotes and
amastigotes of Trypanosoma cruzi. Bioorg Med Chem Lett. 2004;14:3555–8. https://
doi.org/10.1002/ptr.2676.
[111] Venugopal DVR, Yarla NS, Umadevi P. Synthesis, of Novel Piperine Analogs of
Dipeptidyl Boronic Acid as Antimicrobial and Antican-cer Agents. Med Chem. 2014;
4:606–10. https://doi.org/10.4172/2161-0444.1000201.
[112] Sangwan PL, Koul JL, Koul S, Reddy MV, Thota N, Khan IA, et al. Piperine analogs as
potent Staphylococcus aureus NorA efflux pump inhibitors. Bioorg Med Chem. 2008;
16:9847–57. https://doi.org/10.1016/j.bmc.2008.09.042.
[113] Ramesh A. P. Sundarraj, Dr. J. Balamani Synthesis And Biological Evaluation Of 5-(3,4-
Methylenedioxyphenyl)-2,4-Pentadienoic Acid Derivatives As Potent Antimicrobial
And Cytotoxicity Agents, IOSR Journal of Applied Chemistry, 11, (6) Ver. I may
2018; 51-57. DOI: . https://doi.org/10.9790/5736-1105015157
[114] Nagai J, Imamura M, Sakagami H, Uesawa Y. QSAR Prediction Model to Search for
Compounds with Selective Cytotoxicity Against Oral Cell Cancer. Medicines. 2019;6:
45. https://doi.org/10.3390/medicines6020045.
[115] Samykutty A, Shetty AV, Dakshinamoorthy G, Bartik MM, Johnson GL, Webb B, et al.
Piperine, a bioactive component of pepper spice exerts therapeutic effects on androgen
dependent and androgen independent prostate cancer cells. PLoS One. 2013;8:e65889.
https://doi.org/10.1371/journal.pone.0065889.
[116] Hegeto LA, Caleffi-Ferracioli KR, Nakamura-Vasconcelos SS, de Almeida AL, Baldin
VP, Nakamura CV, et al. In vitro combinatory activity of piperine and anti-tuberculosis
drugs in Mycobacterium tuberculosis. Tuberculosis. 2018;111:35–40. https://doi.org/
10.1128/AAC.02520-18.
[117] Philipova I, Valcheva V, Mihaylova R, Mateeva M, Doytchinova I, Stavrakov G. Syn-
thetic piperine amide analogs with antimycobacterial activity. Chem Biol Drug Des.
2018;91:763–8. https://doi.org/10.1111/cbdd.13140.
[118] Sharma S, Kumar M, Sharma S, Nargotra A, Koul S, Khan IA. Piperine as an inhibitor of
Rv1258c, a putative multidrug efflux pump of Mycobacterium tuberculosis. J
Antimicrob Chemother. 2010;65:1694–701. https://doi.org/10.1093/jac/dkq186.
[119] Lin Z, Hoult JRS, Bennett DC, Raman A. Stimulation of mouse melanocyte proliferation
by Piper nigrum fruit extract and its main alkaloid, piperine. Planta Med. 1999;65:
600–3. https://doi.org/10.1055/s-1999-14031.
[120] Faas L, Venkatasamy R, Hider RC, Young AR, Soumyanath A. In vivo evaluation of pip-
erine and synthetic analogues as potential treatments for vitiligo using a sparsely
pigmented mouse model. Br J Dermatol. 2008;158:941–50. https://doi.org/10.
1111/j.1365-2133.2008.08464.x.
[121] Elnaggar YSR, Etman SM, Abdelmonsif DA, Abdallah OY. Intranasal piperine-loaded
chitosan nanoparticles as brain-targeted therapy in Alzheimer’s disease: optimization,
biological efficacy, and potential toxicity. J Pharm Sci. 2015;104:3544–56. https://
doi.org/10.1002/jps.24557.
[122] Pachauri M, Gupta ED, Ghosh PC. Piperine loaded PEG-PLGA nanoparticles: Prepara-
tion, characterization and targeted delivery for adjuvant breast cancer chemotherapy.
20
Medicine in Drug Discovery 7 (2020) 100027
J Drug Deliv Sci Technol. 2015;29:269–82. https://doi.org/10.1016/j.jddst.2015.08.
009.
[123] Bhalekar MR, Madgulkar AR, Desale PS, Marium G. Formulation of piperine solid lipid
nanoparticles (SLN) for treatment of rheumatoid arthritis. Drug Dev Ind Pharm. 2017;
43:1003–10. https://doi.org/10.1080/03639045.2017.1291666.
[124] Shaikh J, Ankola DD, Beniwal V, Singh D, Kumar MNVR. Nanoparticle encapsulation
improves oral bioavailability of curcumin by at least 9-fold when compared to
curcumin administered with piperine as absorption enhancer. Eur J Pharm Sci.
2009;37:223–30. https://doi.org/10.1016/j.ejps.2009.02.019.
[125] Moorthi C, Krishnan K, Manavalan R, Kathiresan K. Preparation and characterization
of curcumin–piperine dual drug loaded nanoparticles. Asian Pac J Trop Biomed.
2012;2:841–8. https://doi.org/10.1016/S2221-1691(12)60241-X.
[126] Jain S, Meka SRK, Chatterjee K. Engineering a piperine eluting nanofibrous patch for
cancer treatment. ACS Biomater Sci Eng. 2016;2:1376–85. https://doi.org/10.1021/
acsbiomaterials.6b00297.
[127] Elnaggar YSR, Etman SM, Abdelmonsif DA, Abdallah OY. Novel piperine-loaded
Tween-integrated monoolein cubosomes as brain-targeted oral nanomedicine in
Alzheimer’s disease: pharmaceutical, biological, and toxicological studies. Int J
Nanomedicine. 2015;10:5459. https://doi.org/10.2147/IJN.S87336.
[129] Shoba G, Joy D, Joseph T, Majeed M, Rajendran R, Srinivas P. Influence of piperine on
the pharmacokinetics of curcumin in animals and human volunteers. Planta Med.
1998;64:353–6. https://doi.org/10.1055/s-2006-957450.
[130] Patel S, Devada S, Patel H, Patel N, Bhavsar S, Thaker A. Influence of co-administration
of piperine on pharmacokinetic profile of gatifloxacin in layer birds. Glob Vet. 2011;7:
427–32.
[131] Jafri A, Siddiqui S, Rais J, Ahmad MS, Kumar S, Jafar T, et al. Induction of apoptosis by
piperine in human cervical adenocarcinoma via ROS mediated mitochondrial pathway
and caspase-3 activation. EXCLI J. 2019;18:154. https://doi.org/10.17179/excli2018-
1928.
[132] Talib W. Regressions of breast carcinoma syngraft following treatment with piperine in
combination with thymoquinone. Sci Pharm. 2017;85:27. https://doi.org/10.3390/
scipharm85030027.
[133] Siddiqui S, Ahamad MS, Jafri A, Afzal M, Arshad M. Piperine triggers apoptosis of
human oral squamous carcinoma through cell cycle arrest and mitochondrial oxidative
stress. Nutr Cancer. 2017;69:791–9. https://doi.org/10.1080/01635581.2017.
1310260.
[134] Ba Y, Malhotra A. Potential of piperine in modulation of voltage-gated K+ current and
its influences on cell cycle arrest and apoptosis in human prostate cancer cells. Eur Rev
Med Pharmacol Sci. 2018;22:8999–9011. https://doi.org/10.26355/eurrev_201812_
16671.
[135] Gunasekaran V, Elangovan K, Devaraj SN. Targeting hepatocellular carcinoma with
piperine by radical-mediated mitochondrial pathway of apoptosis: an in vitro and in
vivo study. Food Chem Toxicol. 2017;105:106–18. https://doi.org/10.1016/j.fct.
2017.03.029.
[136] Greenshields AL, Doucette CD, Sutton KM, Madera L, Annan H, Yaffe PB, et al. Piperine
inhibits the growth and motility of triple-negative breast cancer cells. Cancer Lett.
2015;357:129–40. https://doi.org/10.1016/j.canlet.2014.11.017.
[137] Do MT, Kim HG, Choi JH, Khanal T, Park BH, Tran TP, et al. Antitumor efficacy of pip-
erine in the treatment of human HER2-overexpressing breast cancer cells. Food Chem.
2013;141:2591–9. https://doi.org/10.1016/j.foodchem.2013.04.125.
[138] Yoo ES, Choo GS, Kim SH, Woo JS, Kim HJ, Park YS, et al. Antitumor and Apoptosis-
inducing Effects of Piperine on Human Melanoma Cells. Anticancer Res. 2019;39:
1883–92. https://doi.org/10.21873/anticanres.
[139] Sharma S, Kalia NP, Suden P, Chauhan PS, Kumar M, Ram AB, et al. Protective efficacy
of piperine against Mycobacterium tuberculosis. Tuberculosis. 2014;94:389–96.
https://doi.org/10.1016/j.tube.2014.04.007.
[140] Jin J, Zhang J, Guo N, Feng H, Li L, Liang J, et al. The plant alkaloid piperine as a po-
tential inhibitor of ethidium bromide efflux in Mycobacterium smegmatis. J Med
Microbiol. 2011;60:223–9. https://doi.org/10.1099/jmm.0.025734-0.
[141] Rukachaisirikul T, Prabpai S, Champung P, Suksamrarn A. Chabamide, a novel piper-
ine dimer from stems of Piper chaba. Planta Med. 2002;68:853–5. https://doi.org/10.
1055/s-2002-34410.
[142] Raja A, Kapur A, Fijju M, Mohamed Salique S. In vitro studies on Efflux pump Inhibi-
tion of Catharanthus roseus and piperine against ofloxacin resistant M. tuberculosis. Int
J Pharm Sci Invent. 2015:4.
[143] Piyachaturawat P, Glinsukon T, Peugvicha P. Postcoital antifertility effect of piperine.
Contraception 1982;26:625–33, Piyachaturawat P, Glinsukon T, Peugvicha P. Postcoi-
tal antifertility effect of piperine. Contraception. 1982;26:625–33.
[144] Johri RK, Thusu N, Khajuria A, Zutshi U. Piperine-mediated changes in the perme-
ability of rat intestinal epithelial cells: the status of γ-glutamyl transpeptidase ac-
tivity, uptake of amino acids and lipid peroxidation. Biochem Pharmacol. 1992;
43:1401–7.
[145] Atal CK, Dubey RK, Singh J. Biochemical basis of enh-anced drug bioavailability by
piperine: evidence that piperine is a potent inhibitor of drug metabolism. J Pharmacol
Exp Ther. 1985;232:258–62.
[146] Lakshmipati A. Onehandred Useful Drugs. 3rd ed.. Madras, India: Arogya Ashram
Samithi; 1946.
[147] Lele RD. Ayurveda through modern eyes. Ayurveda Mod. Med. Bombay: Bharatiya
Vidya Bhavan; 1986.
[148] Johri RK, Zutshi U. An Ayurvedic formulation ‘Trikatu’and its constituents. J
Ethnopharmacol. 1992;37:85–91. https://doi.org/10.1016/0378-8741(92)90067-2.
[149] Anonymous. The Ayurvedic Formulary of India Part-II. first English ed.. New Delhi:
Government of India, Ministry of Health and Family Welfare, Department of Indian
Systems of Medicine & Homeopathy; 2000.
[150] Nadkarni KM. Piper nigrum L. Indian Materia Medica. Bombay: Popular Prakashan
Press; 1986. p. 969–72.
A. Tiwari et al.
[151] Usmanghani K, Saeed A, Alam MT. Indusyunic Medicine. Karachi: University of Kara-
chi Press; 1997.
[152] Singh RK, Banerjee R, Upadhyay S, Mitra A, Hazra J. Toxicological evaluation of
Panchakola Avaleha, an Ayurvedic classical formulation, in albino rats. Ayu. 2012;
33:303. https://doi.org/10.4103/0974-8520.105257.
[153] Soni A, Patel K, Gupta SN. Clinical evaluation of Vardhamana Pippali Rasayana in the
management of Amavata (Rheumatoid Arthritis). Ayu. 2011;32:177. https://doi.org/
10.4103/0974-8520.92555.
[154] Gyawali S, Khan GM, Lamichane S, Gautam J, Ghimire S, Adhikari R, et al. Evaluation
of anti-secretory and anti-ulcerogenic activities of avipattikar churna on the peptic ul-
cers in experimental rats. J Clin Diagnostic Res JCDR. 2013;7:1135. https://doi.org/
10.7860/JCDR/2013/5309.3058.
[155] Zaveri M, Khandhar A, Patel S, Patel A. Chemistry and pharmacology of Piper longum
L. Int J Pharm Sci Rev Res. 2010;5:67–76.
[156] Ngane AN, Biyiti L, Bouchet PH, Nkengfack A, Zollo PHA. Antifungal activity of Piper
guineense of Cameroon. Fitoterapia. 2003;74:464–8. https://doi.org/10.1016/S0367-
326X(03)00112-6.
[157] Dzoyem JP, Kuete V. Review of the antifungal potential of African medicinal plants.
Antifung. Metab. from Plants. Springer; 2013; 79–153. https://doi.org/10.1007/978-
3-642-38076-1_4.
[158] Dzoyem JP, Tchuenguem RT, Kuiate JR, Teke GN, Kechia FA, Kuete V. In vitro and in
vivo antifungal activities of selected Cameroonian dietary spices. BMC Complement
Altern Med. 2014;14:58. https://doi.org/10.1186/1472-6882-14-58.
[159] Ebana RUB, Edet UO, Ekanemesang UM, Ikon GM, Etok CA, Edet AP. Antimicrobial ac-
tivity, phytochemical screening and nutrient analysis of Tetrapleura tetraptera and
Piper guineense. Asian J Med Heal. 2016:1–8. https://doi.org/10.9734/AJMAH/
2016/29362.
21
Medicine in Drug Discovery 7 (2020) 100027
[160] Mgbeahuruike EE, Holm Y, Vuorela H, Amandikwa C, Fyhrquist P. An ethnobotanical
survey and antifungal activity of Piper guineense used for the treatment of fungal infec-
tions in West-African traditional medicine. J Ethnopharmacol. 2019;229:157–66.
https://doi.org/10.1016/j.jep.2018.10.005.
[161] Sharangdhara samhitha Madhyamakhanda 6/130 , AFI.
[162] Vora A, Patel S, Patel K. Role of Risorine in the Treatment of Drug Susceptible Pulmo-
nary Tuberculosis: A Pilot Study. J Assoc Physicians India. 2016;64:20–4 PMID:
27805329.
[163] Ansari KA, Akram M. Filfil Siyah (Piper nigrum Linn) an important Drug of Unani Sys-
tem of Medicine: A Review. J Pharmacogn Phytochem. 2014;2:219–21.
[164] Bashir F, Afrin Z. Zanjabeel (Zingiber offcinale) Transformation of Culinary Spice to a
multi-functional Medicine. J Drug Deliv Ther. 2019;9:721–5.
[165] Ahmad B, Akhtar J. Unani system of medicine. Pharmacogn Rev. 2007;1:210.
[166] Afzal M, Khan NA, Ghufran A, Iqbal A, Inamuddin M. Diuretic and nephroprotective
effect of Jawarish Zarooni Sada—a polyherbal unani formulation. J Ethnopharmacol.
2004;91:219–23.
[167] Ganga B, Wadud A, Jahan N, Makbul SAA. Anti-inflammatory and analgesic activity of
Habbe Gule Aakh, A polyherbal Unani formulation in animal models. J Ayurveda
Integr Med. 2018.
[168] Jeganathan NS, Anbazhagan S, Aruna P, Deepa D, Kalaivani A. Preparation and quality
control of a Siddha formulation–Kesari Lehyam. Int J Adv Res Biol Sci. 2016;3:180–5.
[169] Riyasdeen A, Periasamy VS, Paul P, Alshatwi AA, Akbarsha MA. Chloroform Extract of
Rasagenthi Mezhugu, a Siddha Formulation, as an Evidence-Based Complementary
and Alternative Medicine for HPV-Positive Cervical Cancers. Evidence-Based Comple-
ment Altern Med. 2012;2012.