Chemistry of natural products

1. ACTIVE CONSTITUENT OF CERTAIN CRUDE
DRUGS USED IN INDIGENOUS SYSTEM –LIVER
DYSFUNCTION – Phyllanthus niruri
NAME-PRIYA BARDHAN RAY
ROLL NO-05
STREAM:- MASTER OF PHARMACY IN PHARMACEUTICAL
CHEMISTRY
SEMESTER-1ST
SUBJECT:- CHGEMISTRY OF NATURAL PRODUCTS

2. INTRODUCTION
• WHAT IS INDIGENOUS SYSTEM?
• An indigenous system is a natural form of medicine outside the stream of Western or allopathic medicine practiced by majority of doctors all over the
world today. Indigenous systems have existed since time immemorial, but few of them have been developed into fully-fledged working systems. Among
the most prominent of them are ayurveda, homeopathy, yoga & naturopathy, siddha, unani. Some of these, such as yoga & ayurveda, are ancient medical
system in India, some like homeopathy are highly symptomatic, some like yoga depend on exercise and meditation as methods of treatment, and some,
like naturopathy. avoid any artificial aid. Thus, these systems have served as inspiration for drug discovery.
• WHAT IS LIVER?
• Liver is the heaviest gland & second largest organ of the human body.
• Primary functions of liver include Detoxification, Energy conversion, Immunity, Hormonal balance, Digestion, Fat regulation,
Synthesis of Albumin, α and β globulins,coagulation factors, Storage of fluids, vitamins, minerals etc.
• Liver, the key organ of metabolism and excretion has an immense task of detoxification of xenobiotics, environmental pollutants and
chemotherapeutic agents. Hence, this organ is subjected to variety of diseases and disorders.
• WHAT IS LIVER DYSFUNCTION?
• Liver dysfunction is a condition in which liver is not working efficiently or overloaded with toxins.
• Other diseases of liver include, Hepatocellular diseases (viral hepatitis, ALD), Cholestatic disease (intra and extra hepatic obstruction)
Cirrhosis, Cancer (secondary or primary) , Fatty Liver, Genetic Disorders :-Hemochromatosis (iron storage) & Wilsons disease

3. PHYLLANTHUS
• INTRODUCTION
• Phyllanthus is a large genus of shrubs, trees and rare herbs of the family Euphorbiaceae,
comprising more than 600 species, of which,
• P. accuminatus, P. amarus, P. pulcher, P. niruroides, P. anisolobus, P. orbiculatus, P. emblica, P.
oxyphyllus, P. flexuosus, P. raticulatus, P. fraternes, P. simplex, P. mullernus, P. urinaria, P.
mytrifolis, P. virgatus, P. niruri and P. watsonii were investigated for their phytochemical and
pharmacological properties.
• The genus is found in almost over all warmer parts of the world
• In this topic P. niruri will be discussed.

4. Phyllanthus niruri
• INTRODUCTION:- It is a small erect annual herb growing up to 30–40 cm in height and is indigenous to the Amazon
rainforest and other tropical areas, including South East Asia, Southern India and China.It is known since the time of “Charaka”,
because it is known to be useful for various liver disorder.
• SYNONYMS:- Phyllanthus, Bhui-Amla, Bhūmī āmalakī, Dukong anak, Chanca Piedra, Gale of the wind, Stonebreaker,
Seed-under-leaf.
• BIOLOGICAL SOURCE:- It consists of all aerial parts of the plant, Phyllanthus niruri belonging to family
Euphorbiaceae. Indian variety available is Phyllanthus fraternus and Phyllanthus armarus. It contains not less than 0.25 per
cent of total phyllanthin and hypophyllanthin on dried basis.
• GEOGRAPHICAL SOURCE:- The plant is found growing abundantly throughout India, mainly in the states of
Maharashtra, Uttar Pradesh, Punjab, Bihar, Orissa, Andhra Pradesh and some parts of Madhya Pradesh, Karnataka and Bengal.
Also found in Sri Lanka.
• CULTIVATION & COLLECTION:- It grows well under tropical climatic conditions. However, very dry temperature
or very low temperature conditions are not preferred. It can grow up to 700 - 800 metres above the sea level. Suitable
temperature ranging from 25 - 40°C with average humidity is desired. Well distributed rainfall in the range of 40 - 50 cm is
suitable. Bhuiamla grows in variety of soils, preferably well drained rich organic and light textured soils. It also grows well on
sandy loam to calcareous soil. Soil pH should be alkaline to acidic. Even water logging soil does not show adverse effects on its
growth. Propagated well through seeds or by raising seedlings and transplanting. The plants mature within 3 to 4 months. They
are usually harvested after rainy season is over, when the amount of green leaves is in abundance. Since, the active constituent
is present in leaves, higher leaf mass is expected during harvesting. For harvesting, the whole plants are uprooted. The collected
whole plants are cleared of external matter and are allowed to dry in shade. The fresh juice of plant is also used in medicine.
Total yield of fresh herb per hectare is about 2 - 3 tones.

5. Phyllanthus niruri
• MACROSCOPIC CHARACTER:-
o COLOUR- Green to yellowish green
in colour
o TASTE- Bitter
o ODOUR- None
o STEMS- 1 - 1.5 cm in length and 1.4
mm in diameter stems are stout.
o LEAVES- Short-stalked, oblong in
shape and about 5 × 3 mm in size.
• PHOTOGRAPH OF PLANT

6. EXTRACTION METHOD OF Phyllanthus niruri
Fresh plants
The plant was dried and grinded into fine powder
Hundred gram of the powder was soaked in 1000 mL of 95% ethanol for 3 days
Then, the mixture was filtered using filter paper (Whatman No. 1)
Distilled under reduced pressure in a rotating evaporator (Buchi, Switzerland).
The ethanol extract was then dissolved in 10% Tween 20.

7. ACTIVE CHEMICAL CONSTITUENTS & TYPES OF
PHYTOCHEMICALS
• Many of the active constituents to which the biological activity of P. niruri has been attributed
include lignans, tannins, coumarins, terpenes, flavonoids, alkaloids, saponins &
phenylpropanoids.
• These are found in the leaves, stems & roots of this plants.
• Common lipids, sterols & flavonols also occur in the plant.
• It contains, leucodelphimidin alkaloids, flavonoids as quercetin, astralgin, nerurin quercitrin,
isoquercitrin and rutin. It also contains |< 0.25 percent of total phyllanthine and
hypophyllanthine. Mainly these two lignans are responsible for hepatoprotective & anti-
hepatitis activity
• Niruriside, a carbohydrate from methanolic extract of phyllunthus niruri has shown anti-HIV
activity. It is specific inhibitor of REV protein, with an IC50 value of 3.3 µm.

8. Types of chemicals found in the plant
CHEMICAL CLASS PHYTOCHEMICALS PRESENT PHARMACOLOGICAL EFFECT
FLAVONOIDS Rutin, Quercetin, Quircitin, Astragalin, Catachin Blood vessel strengthener, Anti-oxidant, Anti-
Cancer, Anti-inflammatory, Antiseptic, Anti-
leishmanial, Antidiarrhoeal, Diuretics.
TERPENES LIMONENE, P-cymene, LUPEOL Anticarcinogenic, Antioxidant,
COUMARINS Ellagic Acid, Methyl Brevifolincarboxylate Chemopreventive effect in cancer, Vasorelaxent
TANNINS Repandusinic Acid, Geraniin, Corilagin Reverse transcriptase inhibitor, Anti-hypertensive,
Anti-fungal, Thrombolytics.
SAPONINS Diosgenin Anti-fungal, Cardiovascular active
LIGNANS Phyllanthin, Hypophyllanthin, Niranthin,
Nirtetralin, Phyltetralin, Linletralin, Isolintetralin,
Cubebin, Urinatetralin
Hepatoprotective, Antigenotoxic, Anti-HIV, Anti-
inflammatory, Anti-viral, Anti-tumour
FLAVANONE
GLYCOSIDE
Nirurin, Niruriflavone Radical scavenging properties.
ALKALOIDS Norsecuririne, Nirurine, Phyllanthine,
Phyllochrysine, Securirine
Anti-spasmodic, Anti-malarial, Anti-microbial, Anti-
bacterial, Selective GABA antagonist
CARBOHYDRATE Niruriside Anti-HIV, specific inhibitor of REV protein

9. Structure of Lignans
PHYLLANTHIN HYPOPHYLLANTHIN
SAR of Phyllanthin

10. Phyllanthus niruri IN LIVER DYSFUNCTION
• Phyllanthus primarily contains lignans (e.g., phyllanthine and hypophyllanthine) & have been
shown to possess hepatoprotective activity.
• Phyllanthus niruri has a very long tradition of use as cure for liver diseases as it was mentioned
as one of herbs for treatment of jaundice in ancient texts of Ayurveda. Many laboratory and
clinical tests has proved niruri's effectiveness of protecting liver from damage (by alcohol,
chemical toxines, drugs such as Paracetamol, Nimesulide, CCl4 induced liver toxicity,
galactosamine induced cytotoxicity), in tonifying and strenghtening liver, balancing liver
enzymes, and killing pathogens that infect liver (including hepatitis B virus, Staphylococcus
aureus).
• Drug-induced liver injury is a major health problem that challenges not only health care
professionals but also the pharmaceutical industry and drug regulatory agencies.
• The Carbon tetrachloride and Galactosamine induced cytotoxicity can be decreased by the P.
niruri hexane extract. Phyllanthin and hypophyllanthin protects against the CCL4- &
Galactosamine induced cell lesions and GalN induced Hepatotoxicity in liver transferase
enzymes levels & significantly increased protein levels. Also, the protein isolated from this plant
protects the liver from Carbon tetrachloride induced hepatotoxicity and this can be measured
by the liver enzymes and reduced levels of antioxidant enzymes.

11. Phyllanthus niruri IN LIVER DYSFUNCTION
• Phyllanthus niruri can reduce Nimesulide induced hepatic damage. By measuring the levels of glutamate oxaloacetate transaminase
(GOT), glutamate pyruvate transaminase (GPT) and alkaline phosphatase (ALP) in serum it was concluded that the levels of three
enzymes are decreased in the extract treated group. By these observations intra peritoneal treatment was found to be more effective
than oral administration.
• The over dose of Paracetamol leads to hepatotoxicity same as viral infection. The glutamic pyryvic transaminase (GPT) levels of
serum were decreased in the P. niruri extract administration. The serum glutamate pyruvate transaminase (SGPT) and serum glutamate
oxaloacetate transaminase (SGOT) was decreased in the invivo studies. The ethanol extract and hexane extract were administered and
the serum parameters (serum bilirubin, serum alkaline phosphatase, serum aspartate (AST), serum alanine transferase (ALT), hepatic
reduced glutathione (GSH) were analysed and these parameters were controlled after the treatment with hexane extract and .hence, it
was stated that P.niruri can control the paracetamol induced hepatotoxicity.
• Protein isolated from this plant was found to enhance cell viability against Tertiary butyl hydroperoxide induced cytotoxicity and cell
death; and it protects hepatocytes against Thioacetamide induced cytotoxicity. The extract prevents the alterations in GSH levels and it
also reduces the lipid peroxidation induced by TAA.
• Alcohol is a toxin in higher doses and when it is associated with poly unsaturated fatty acids (PUFA) induces oxidative stress &
hepatotoxicity. This can be efficiently reduced by P.niruri extract analyzed by the antioxidant potentials of liver enzymes and
histopathological studies.
• In alcoholic extracts treated groups there was statistical significant decrease in the levels of serum bilirubin, SGOT, SGPT and ALP as
compared to the hepatotoxic group. In significant reduction in the activity levels of superoxide dismutase, catalase, glutathione
peroxidase, gluthaione-s-transferase, and also glutathione reductase content were observed in the liver of S. aureus intoxicated rats
over controls. Thus, the results suggest the beneficial role of Phyllanthus niruri might be due to its antioxidative properties against S.
aureus infection induced oxidative stress in liver.

12. Phyllanthus.niruri has been used to inhibit the
hepadnaviridae extensively for hepatitis B virus.
• MECHANISM OF ACTION:-
• Phyllanthus niruri has been used to inhibit the hepadna virus and it is extensively used to
treat jaundice and hepatitis B virus. It inhibits hepatitis B virus by inhibiting 50 % of
DNA polymerase. Hepatitis B is the most prominent disease in emerging era. Phyllanthus
niruri extract can prevent Hepatitis B by binding to the endogenous DNA polymerase and
even it can bind to the hepatitis B surface antigen in in vitro.
• DNA polymerase is the one of the enzyme which is responsible for the synthesis of DNA
molecule and after the inhibition of these enzyme there is no formation of the DNA
molecule of the virus. So the net result is multiplication of these virus totally affected.
• When the phyllanthin binds on the hepatitis B surface antigen damage, of cell wall of the
virus is prominently occur by this mechanism phyllanthin act as anti- Hepatitis agent.

13. Marketed formulations
• It has been used as a cure since ancient
times in Ayurveda, Siddha, Unani and other
traditional systems of medicine.
• This herb is available on the market in form
of whole dried herb, tea sachettes, powder,
capsules, tablets, and liquid alcoholic and
non-alcoholic extracts. And is still
commonly gathered from wild, but more
and more plantations is now established.
Whole fresh plants are used in the same
way as dried ones.
• The active compounds phyllanthin and
hypophyllanthin isolated from P. niruri are
major components of many popular liver
tonics in India including Liv.-52.

14. REFERENCES
1. Kokate C.K., Purohit A.P., Gokhale S. B. ‘PHARMACOGNOSY’, Edition 43rd, 2009, Nirali Prakashan,
Page No:-13.89-13.91
2. G. Bagalkotkar, S. R. Sagineedu, M. S. Saad and J. Stanslas, ‘Phytochemicals from Phyllanthus niruri
Linn. and their pharmacological properties: a review’, Journal of Pharmacy and Pharmacology, 2006,
58: 1559–1570, https://www.researchgate.net/publication/6475974
3. K. Narendra, J. Swathi, K. M. Sowjanya, A. Krishna Satya, ‘Phyllanthus niruri: A Review on its Ethno
Botanical, Phytochemical and Pharmacological Profile’, Journal of Pharmacy Research, 2012, 5(9),
4681-4691, https://www.researchgate.net/publication/259758390
4. V. Ramamurthy & T. Abarna, ‘Hepatoprotective Activity Of Phyllanthus Niruri Whole Plant Extract
Against Staphylococcus Aureus Intoxicated Albino Rats’ , Global Journal of Biology, Agriculture &
Health Sciences, 2014, Vol. 3(3), 256-260. https://www.longdom.org/archive/gjbahs-volume-3-issue-3-
year-2014.html
5. Zahra A. Amin, Mehmet Bilgen, Mohammed A. Alshawsh, Hapipah M. Ali, A. Hamid A. Hadi,3 and
Mahmood A. Abdulla, ‘Protective Role of Phyllanthus niruri Extract against Thioacetamide-Induced
Liver Cirrhosis in Rat Model’, Evidence-Based Complementary and Alternative Medicine, vol. 2012,
Article ID 241583, 9 pages, 2012. doi:10.1155/2012/241583