CARDIAC GLYCOSIDES

BY
PHARM. EVANS PAUL KWAME AMEADE
DEPT. OF PHARMACOGNOSY AND HERBAL MEDICINE
SCHOOL OF PHARMACY AND PHARMACEUTICAL SCIENCES
UNIVERSITY FOR DEVELOPMENT STUDIES
PHARM.D, LEVEL 300
FEVRUARY, 2023
• Outline
• What are cardiac glycosides
• Sources of cardiac glycosides
• Structure of cardiac glycosides
• Biogenesis of cardiac glycosides
• Plant drug containing cardiac glycosides
• Test for cardiac glycosides
What are cardiac glycosides
• Naturally occurring secondary metabolites which in
appropriate small quantities has beneficial effect on the
heart but can become toxic in high concentration.
• Plants containing cardiac steroids have been used as poisons
and as cardiotonic agents which are widely used in the
modern treatment of congestive heart failure and for
treatment of atrial fibrillation and flutter
Congestive heart failure
• Heart diseases can be primarily grouped into three major disorders: cardiac
failure, ischemia and cardiac arrhythmia.
• Cardiac failure can be described as the inability of the heart to pump blood
effectively at a rate that meets the needs of the body because the muscles of
the heart especially the ventricles which are to contract to force blood out of
the heart are performing weakly.
• Reduced contraction of heart leads to reduced heart output but new blood
keeps coming in resulting in the increase in heart blood volume. The heart
feels congested. Hence the term congestive heart failure (CHF).
• CHF leads to lowered blood pressure and poor renal blood flow which results
in oedema in the lower extremities and the lung (pulmonary oedema).
• Renal failure ultimately occurs in unmanaged CHF.
Sources of cardiac glycosides (CardG)
• > 400 naturally occurring cardiac glycosides exist
• CardG also occur in only some genera in plant families such as
• Apocynaceae (Apocynum, Nerium, Strophantus, Thevetia)
• Scrophularaceae (Digitalis)
• Liliaceae (Convallaria, Urginea)
• Ranunculaceae (Adonis, Helleborus)
• Crassulaceae (Cotyledon, Kalanchoe)
• Brassicaceae (Erysimum)
• Celastraceae (Euonymus)
• Asclepiadaceae (Asclepias, Calotropis, Cryptostegia, Periploca,
Xysmalobium)
• CardG occurs in several parts of the plants including the seeds,
leaves, stems, roots or barks.
• Notable plants that are known to produce cardiac glycosides
are
• Common oleander (Nerium oleander)
• Yellow oleander (Thevetia peruviana)
• Purple Foxglove (Digitalis purpurea)
• Lily of the valley (Convallaria majalis)
• Kalanchoe (Bryophyllum spp.)
• Azalea (Rhododendron spp.)
• Woolly foxglove (Digitalis lanata)
• Ouabain (Strophanthus gratus)
• Squill bulb (Drimia maritima [syn. Urginea maritima])
• Kombe arrow poison (Strophanthus kombe)
Common oleander Yellow oleander

Lily of the valley Kalanchoe Azalea

Structure and types of cardiac glycosides
• The basic structure of cardiac glycosides is composed of
• a sterol nucleus with an unsaturated conjugated lactone ring
attached to the C17 position,
• a hydroxyl group at the C3 position of the nucleus with a sugar.
• The basic structure of the steroidal part is cyclopentano
perhydrorphenanthrene nucleus, to which a lactone ring is
attached
• Other important features in the structure include
• 14 β – OH group at C – 14
• The four rings of the steroidal nucleus are fused differently
• B/C ring is trans,
• C/D ring and A/B ring are mostly cis
• Such ring fusion gives the aglycone nucleus of cardiac glycosides the
characteristic 'U' shape

U-shape of the aglycone molecule of cardiac glycoside

 • Additional OH groups at C-5, C – 11 and C – 16 may be present
• Depending on the type of unsaturated lactone ring attached
to the C17 position of the cardiac glycoside, they are
classified into two types
• type A (a five-membered unsaturated lactone ring)
• type B (a six-membered unsaturated lactone ring).
• The type A (a five-membered unsaturated lactone ring) is called the
Cardenolide
• The type B (a six-membered unsaturated lactone ring) is referred to as
Bufadienolide
• Plants and animals such as toads (Bufonidae) and some insects can
produce both cardenolides and bufadienolides.
•Examples of Cardenolide
•Digitalis glycosides
•Digoxin
•Digitoxin
•Gitoxin
•Strophanthus gratus glycoside
•Oubain
•Strophanthus Kombe glycoside :
•K- strophanthin
•Although best studied in leaves, cardenolides can occur in all
plant tissues.
•The latex seem to have the highest concentration of cardenolides in
some plant species.
Digoxin

Gitoxin Digitoxin
Oubain K- strophanthin
• Example of bufadienolide:
• Squill bulb glycoside
• Scillaren
• Venenum bufonis
• bufalin, cinobufagin, and resibufogenin
• One to 4 sugars are found to be present in most cardiac
glycosides attached to the 3 β-OH group.
• The glycone portions determine the pharmacodynamic and
pharmacokinetic activities of each cardiac glycoside but possess
no biological activity
• The commonly found sugars include
• Glucose, Galactose, Mannose, Rhamnose, Digitalose, Digitoxose,
Digginose, Vallarose, Fructose
• The most common monosaccharides found in cardiac glycosides
are however glucose, rhamnose and 6-deoxy monosaccharides
• Whenever glucose is part of the glycone, it is always at the terminal
end
Glucose at the terminal
position of the
glycone unit
• The names of the aglycone are derived from the name
of the cardiac glycosides which are commonly named
after the plant from which they occur.
• The term 'genin' at the end refers to only the aglycone
portion of a cardiac glycoside (there are exceptions)
• From the Digitalis purpurea comes the cardiac
glycoside ‘Digitoxin’ which has the aglycone
‘Digitoxigenin” and three sugar moieties forming the
glycone.
• Other aglycone molecules include
• digoxigenin, gitoxigenin, strophanthidin and bufalin.
• The "backbone" U shape of the steroid nucleus appears to be
very important since sugar molecules and lactone alone do
not have any activity.
• The lactone ring even if joined to the steroid molecule is not
absolutely required since its replacement with unsaturated
nitrile (C=C-CN group) found the cardiac glycoside to have
little or no loss in biological activity.
• Structures with C/D trans fusion are inactive whilst
conversion of A/B from cis to trans system leads to a marked
drop in activity.
• The C14 -OH groups is dispensable since a skeleton without
C14 -OH group but with C/D cis rings fusion remained active.
• Although lactone alone is inactive, the unsaturated 17-
lactone plays an important role in receptor binding since
saturation of the lactone ring dramatically reduced the
biological activity.
• More lipophilic Cardiac glycosides are absorbed faster and
exhibit longer duration of action as a result of slower urinary
excretion rate.
• Lipophilicity is strongly influenced by the number of sugar
residues and the number of hydroxyl groups on the aglycone part
of the glycoside.
• For example, digitoxin and digoxin structures differ only by an
extra OH group in digoxin at C-12, yet their partition coefficients
differ by as much as 15 % points
General characteristics of Cardiac glycoside
• Colourless or white crystals (rarely amorphous)
• Solubility is determined by the number of sugar
moieties; aglycones – soluble in organic solvents,
insoluble in water; but glycosides – soluble in
water and slightly soluble in ethanol and
chloroform
• Optically active compounds
• Undergo hydrolysis (acid, enzymatic and alkaline)
Biogenesis of Cardiac glycosides
• Cardenolides are derived from mevalonic acid via phytosterol
and pregnane intermediates
• The cholesterol is formed by the joining of 3,3-dimethylally
pyrophosphate (DMAPP) or its isomer isopentenyl-
pyrophosphate (IPP) produced in the mevalonate pathway
• Pregnenolone formed by a mitochondrial cytochrome P450-
dependent side chain cleaving enzyme is modified through
several steps to produce the genin.
• Pregnenolone may be considered as the starting point for
cardenolide formation regardless of the assumed sterol
precursor
• The most common sequence of the individual biosynthetic
steps leading to 5β-cardenolides is not yet clear and more
than one pathway may be operative.
Formation of the sterol from DMAPP and IPP
• The aglycones (secondary cardenolides) are glucosylated by
cytoplasmatic glucosyltransferases and actively transported
into the vacuole by a primary glycosidetranslocase in
Digitalis.
• In Digitalis, cardenolides can be interconverted between the
cardiac glycoside (primary cardenolides) the form principally
stored in the cell vacuoles and the aglycone form.
• Glucosidation could enhance the polarity of the cardenolide
and therefore prevent its passive efflux out of the vacuole.
Activity of cardiac glycoside
• Cardiac glycosides affect the heart both directly and indirectly in a
series of complex actions, some of which oppose one another.
• The direct effect is to inhibit the membrane-bound sodium–potassium
adenosine triphosphatase (Na+, K+-ATPase) enzyme that supplies
energy for the system that pumps sodium out of and transports
potassium into contracting and conducting cells.
• The indirect effect is to enhance vagal activity by complex peripheral
and central mechanisms.
• The clinically important consequences are on:
• the contracting cells: increased contractility and excitability
• SA and AV nodes and conducting tissue: decreased impulse generation and
propagation.
Importance of cardiac glycosides to the life
• These same compounds have been applied to poison arrows in
human warfare
• They are used by plants to protect themselves against herbivory
• Some insects sequester cardiac glycosides for protection against their
predators.
Mechanisms of action of cardiac glycosides as drugs
• The most important use of the cardiac glycosides is its effects in
treatment of cardiac failure.
• In cardiac failure, or congestive heart failure, heart cannot pump
sufficient blood to maintain body needs.
• During each heart contraction, there is an influx of Na+ and an
outflow of K+.
• Before the next contraction, Na+, K+‐ATPase must reestablish the
concentration gradient pumping Na+ into the cell against a
concentration gradient. This process requires energy, which is
obtained from hydrolysis of ATP to ADP by Na+, K+‐ATPase.
• Cardiac glycosides inhibit Na+, K+‐ATPase, and consequently
increase the force of myocardial contraction
• By inhibiting the Na+/K+-ATPase, cardiac glycosides cause
intracellular sodium concentration to increase. This then
leads to an accumulation of intracellular calcium via the Na+-
Ca2+ exchange system.
• In the heart, increased intracellular calcium causes more
calcium to be released by the sarcoplasmic reticulum,
thereby making more calcium available to bind to troponin-
C, which increases contractility
(inotropy)
• Cardiac glycosides also increase vagal efferent activity to the
heart, reducing sinoatrial firing rate (causing bradycardia)
and conduction velocity of electrical impulses through the
atrioventricular (AV) node.
• Hence digoxin also used for the the treatment of supra-
ventricular arrhythmias, particularly atrial fibrillation
Plant drug containing Cardiac glycosides
• Some plants are known to contain cardiac glycosides of
commercial value and they include:
• Digitalis - Digitalis purpurea leaves (foxglove), Digitalis lanata leaves -
white flowers
• Strophanthus vine seeds
• Urginea bulbs (squill)
• Convallaria leaves (lily of the valley)
Digitalis
• Digitalis is a genus of about 20 species of
herbaceous, perennials, shrubs and biennials
commonly called foxgloves
• Biological source
• It is obtained from dried leaves of Digitalis purpurea
• Digitalis lanata is another source of cardiac glycosides
Chemical constituents
• Digitalis contain 0.2 to
0.45%
and of both
secondary primary
glycosides.
• Digitalis purpurea
contains
• Primary glycosides:
Purpurea
and B, glycosides
glucogetaloxin A
• Secondary:
gitoxin and digitoxin,
getaloxin.
• Primary
less glycosides
stable and less are
significant
secondary than
glycosides.
• Purpurea
and B glycosides
constitute the A
principal
constituentactive
of the fresh
leaves.
Digitalis lanata contains
• Digitoxin, gitoxin, digoxin.
• Lanatoside A, B, C, D & E
• Lanatoside is acetylated products of purpurea
glycosides
 Digitalis lanata
Digitalis purpurea - Purple foxglove
Strophanthus glycosides
• Strophanthus, which is of the Apocynaceae family, is a
flowering plant that grows in tropical Africa, South
Africa, southern India, the Philippines, Laos, Vietnam,
and South China.
• The name Strophanthus is derived from the Greek
strophos (a twisted cord or rope) and anthos (a
flower).
e.g. Strophanthus kombe, Strophanthus gratus
Chemical constituents
• A mixture of cardiac glycosides isolated from Strophantus kombe
mainly contains K-strophanthin-β and K-strophanthozide.
• K-Strophanthin-β consists of the aglycon of strophanthidin and a
sugar residue made up of cymarose-β-d-glucose. K-
strophantozide has a sugar residue of three units: cymarose-β-d-
glucoso-α-d-glucose.
• Use of the term strophanthin generally refers to all glucosides of
this series
• Strophantus gratus contains the cardioactive glycoside (Ouabain)
or G- strophanthin.
• The seeds are the part of the plant used; used as
arrow poison as it is extremely poisonous
• Indications: Atrial fibrillation, Atrial flutter, Cardiac
arrhythmia, Left ventricular failure, Ventricular
arrhythmia
 Strophantus gratus

Strophantus kombe
Squill
• Contain about 15 glycosides; 0.1% to 2.4% total bufadienolides
• It is the dry fleshy inner scales of the bulb of the white variety
of:Urginea maritima (synonymous to Drimia maritima ) (Fam.
Liliaceae): Mediterranean squill.
• There is also the red squill which is also known as sea onion, is
obtained in the powder form from a plant Urginea indica. : Indian
squill.
• The dried powders of red squill have been used for the control of
rodents since the 13th century.
• Although red squill has many alkaloids, scilliroside is the most toxic
and provides rodenticidal activity.
White squill (Drimia maritima)
Chemical constituents
• White variety:
• average 0.2%-0.4%
• proscillaridin A
• scillaren A
• glucoscillaren A (aglycone: scillarenin),
• scilliphaeoside
• Scilliglaucoside
• Red variety:
• < 0.1% scilliroside and glucoscilliroside (aglycone: scillirosidin);
• proscillaridin A
• scillaren A
Uses
• Squill glycosides have
• similar action to digitalis glycosides, but they have a, more rapid
action (rapid onset of action), but less used.
• diuretic action and
• expectorant.
• The red bulb is used as rat poison and not as cardiac glycoside.
• It kills rats only and not other animals.
• Red squill is 100-500 times more toxic to rats than is the white
variety
Chemical tests for cardiac glycosides:
Raymond’s test:
• To the drug, add a few ml of 50% ethanol and 0.1 ml of 1 % solution of m-
dinitrobenzene in ethanol. To this solution, add 2-3 drops of 20% sodium
hydroxide solution.
• Violet colors appears, this is due to presence of active methylene group.

Legal test:
• To the drug, add few ml of pyridine and 2drops of nitroprusside and a drop
of 20% sodium hydroxide solution.
• A deep red colour is produced.
Keller killiani test:
• Glycoside is dissolved in a mixture of 1 % ferric sulphate solution in (5%)
glacial acetic acid.
• Add one or two drop of concentrated sulphuric acid.
• A blue colour develops due to the presence of deoxy sugar.
Xanthydrol test:
• The crude is heated with 0.1 to 5% solution of Xanthydrol in glacial acetic
acid containing 1% hydrochloric acid.
• A red colour is produced due to the presence of 2-deoxysugar.
Baljet test:
• Take a piece of lamina or thick section of the leaf and add sodium
picrate reagent.
• If glycoside is present yellow to orange colour will be seen.
Kedde test:
• A solution of glycosides is treated with a small amount of Kedde
reagent (Mix equal volumes of a 2% solution of 3, 5 dinitrobenzoic
acid in menthol and a 7.5% aqueous solution of KOH).
• Development of a blue or violet colour that faded out in l to 2 hrs
shows it presence of cardenolides.
Antimony trichloride test:
• To a solution of glycoside add a solution of antimony tri-
chloride and tri-chloroacetic acid, and then heat the
mixture.
• Appearance of blue or violet colour show presence of
cardenolides and bufadienolides
References
• https://www.sciencedirect.com/topics/neuroscience/cardiac-glycosid
es
• http://www.yourarticlelibrary.com/pharmacognosy/photochemical-sc
reening/cardiac-glycosides-types-chemical-test-and-other-details/494
51
• https://www.sciencedirect.com/topics/neuroscience/deoxy-sugars
• https://www.sciencedirect.com/topics/biochemistry-genetics-and-mo
lecular-biology/bufadienolide
• http://www.people.vcu.edu/~urdesai/car.htm
• Kreis, W., Hensel, A., & Stuhlemmer, U. (1998). Cardenolide
Biosynthesis in Foxglove1. Planta Medica, 64(06), 491-499.
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