VINCA ALKALOIDS

Presented by-
JYOTI
SINGH
M. PHARM,
1ST SEMESTER
University Institute of
Pharmaceutical sciences,
Panjab
University, Chandigarh
CONTENTS
 Alkaloids
 Classification of alkaloids
 Vinca alkaloids
 Botanical classification
 History
 Physical characteristics
 Chemical tests
 Chemical constituents
 Chemistry
 Structure Activity Relationship (SAR)
 Biosynthesis
 Isolation
 Purification
 Characterization
 UV Spectrum
 IR Spectrum
 Mass spectrum
 13 C NMR Spectrum
 H NMR Spectrum
 Uses
 Drug interactions
 Side effects
 Recent advancements
 Marketed products
 References
ALKALOIDS
 Alkaloids are a chemically heterogenous group of natural substances and
comprise more than 6000 basic nitrogen containing organic compounds
which occur in about 15% of all vascular terrestrial plants and in more than
150 different plant families.
 The term is derived from the word ‘alkali-like’ and hence, they resemble
some of the characters of naturally occurring complex amines.
 These are the organic products of natural or synthetic origin which are
basic in nature and contain one or more nitrogen atoms, normally of
heterocyclic nature, and possess specific physiological actions on human or
animal body, when used in small quantities.
 They are found primarily in plants but can also occur in animals and fungi.
 Many alkaloids have pharmacological effects and are used in medicine,
while others may be toxic.
Example: Strychnine- A powerful poison that can cause muscle spasms,
rigidity and weakness, leading to respiratory failure and death.
CLASSIFICATION OF ALKALOIDS
 On the basis of their biogenesis:
 On the basis of their chemical nature:
This is the most accepted way of classification of alkaloids. The main criterion
for chemical classification is the type of fundamental (normally heterocyclic)
ring structure present in alkaloid.
The alkaloidal drugs are broadly categorized into two divisions:
1. Heterocyclic alkaloids (True alkaloids)
2. Non-heterocyclic alkaloids or proto-alkaloids or biological amines or
pseudoalkaloids
VINCA ALKALOIDS
 SYNONYMS
Catharanthus, periwinkle
 BIOLOGICAL SOURCE
It is the dried whole plant of Catharanthus roseus, belonging to family
Apocynaceae. It is also known as Vinca rosea.
 GEOGRAPHICAL SOURCE
It is probably indigenous to Madagascar. It is cultivalted in South Africa, India,
U.S.A, Europe, Australia and Caribbean islands as an ornamental plant, as well
as, for its medicinal properties.
 DESCRIPTION
It is an erect, everblooming glabrous, 30 to 60 cm high. The plant has spread
all over the tropical and subtropical parts of India, and grows wild all over the
plains and lower foothills in northern and southern hills of India.
BOTANICAL CLASSIFICATION
HISTORY
 Probably the use of vinca has been known since B.C. 50 in European
countries as antidysentric, antihaemorrhagic, diuretic and wound healing.
 This plant was used in the form of ‘tea’ for treatment of diabetes in Jamaica
and in Brazil for toothache.
 This plant was first scientifically investigated by Canadian workers Noble,
Beer and Cutts. During these studies, it was found that it does not have any
oral hypoglycemic principle, but contains alkaloid possessing antileukemic
principle and the alkaloid was named as vincaleucoblastine.
 Because of such activity, the plant was thoroughly investigated at M/s Eli
Lilly by Svoboda and his colleagues and they reported four dimeric indole
alkaloids, viz. vinca-leucoblastine, leurosine, leurosidine and leurocristine.
All these compounds exhibited anticancer activities.
 Some changes were later reported for the names of these compounds such
as vincaleucoblastine to vinblastine and leurosine, leurosidine and
leurocristine to vinleurosine, vinrosidine and vincristine respectively.
PHYSICAL CHARACTERISTICS
 1. Molecular Weight:
Vinca alkaloids have relatively high molecular weights. For example:
• Vincristine: ~825.98 g/mol
• Vinblastine: ~811.03 g/mol
• Vindesine: ~753.92 g/mol
• Vinorelbine: ~778.96 g/mol
 2. Optical Activity:
Vinca alkaloids are optically active due to their complex chiral structure. This
means they can rotate plane-polarized light.
 3. Melting Point:
The melting points of vinca alkaloids are typically high, reflecting their
crystalline nature:
• Vincristine: Melting point around 218°C (decomposes).
• Vinblastine: Melting point around 267°C (decomposes).
• Vinorelbine: Melting point around 208-210°C.
• Vindesine: Melting point around 205-210°C (decomposes).
 4. Color:
They are typically white or off-white crystalline powders.
 5. Solubility:
They are generally soluble in organic solvents like ethanol, chloroform, and
methanol. Their solubility in water varies, with some vinca alkaloids being
moderately water-soluble.
 6. Chemical Reactivity:
Vinca alkaloids have reactive functional groups, including ester groups, which
make them susceptible to hydrolysis and degradation under certain conditions
(e.g., exposure to moisture, light, or heat).
 7. pH Sensitivity:
Vinca alkaloids are relatively stable at acidic pH but may degrade in more basic
conditions. This is important for their formulation in medicinal products.
CHEMICAL TESTS
 1. Dragendorff’s Test:
• Reagent: Dragendorff’s reagent (a solution of potassium bismuth iodide).
• Procedure: When the reagent is added to an extract of vinca alkaloids, an
orange or reddish-brown precipitate forms, indicating the presence of
alkaloids.
 2. Mayer’s Test:
• Reagent: Mayer’s reagent (potassium mercuric iodide).
• Procedure: When Mayer’s reagent is added to a solution of vinca
alkaloids, a cream-colored precipitate forms, indicating the presence of
alkaloids.
 3. Wagner’s Test:
• Reagent: Wagner’s reagent (iodine in potassium iodide).
• Procedure: On addition of Wagner’s reagent, a reddish-brown precipitate
forms, confirming the presence of alkaloids.
 4. Hager’s Test:
• Reagent: Hager’s reagent (saturated aqueous solution of picric acid).
• Procedure: When Hager’s reagent is added, a yellow crystalline precipitate
forms, indicating the presence of alkaloids.
 5. Vitali-Morin Test:
• Reagent: Fuming nitric acid and alcoholic potassium hydroxide.
• Procedure: Treat the alkaloid extract with fuming nitric acid, followed by
alcoholic potassium hydroxide.
• Positive Result: Development of a violet or purple color.
• Purpose: A specific test for vinca alkaloids based on their indole structure.
 6. Marquis Test:
• Reagent: Formaldehyde and sulfuric acid.
• Procedure: Add sulfuric acid and formaldehyde to the alkaloid extract.
• Positive Result: Development of a red-brown color.
• Purpose: Confirms the presence of vinca alkaloids by generating a color
change specific to their chemical structure.
 7. Sodium Nitroprusside Test:
• Reagent: Sodium nitroprusside solution and ammonia.
• Procedure: Add sodium nitroprusside to the vinca alkaloid solution,
followed by ammonia.
• Positive Result: Formation of a violet color, indicating the presence of
alkaloids.
• Purpose: Confirms alkaloids with amino groups, often present in vinca
alkaloids.
CHEMICAL CONSTITUENTS
 A large number of indole alkaloids are present in vinca. Out of them,
about 20 dimeric indole-dihydroindole alkaloids possess oncolytic
activity, and among them, vincristine and vinblastine are most
significant.
 They contain indole alkaloid part called catharanthine and
dihydroindole alkaloid part called vindoline.
 The other alkaloids present in vinca are ajmalicine, lochnerine,
serpentine and tetrahydroalstonine.
 It requires about 500 kg crude drug to extract out 1 g of vincristine,
because of its extreme low content, viz. 0.0002 %. This makes
these alkaloids very costlier and hence, the efforts for their
synthesis are under attempts.
CHEMISTRY
Vinca alkaloids are indole-dihydroindole alkaloids,
consisting of two distinct structural units:
 Catharanthine: An indole-based moiety.
 Vindoline: A dihydroindole-based moiety.
The two parts are connected by a complex linkage, making
vinca alkaloids unique and complex molecules. The basic
chemical framework of vinca alkaloids consists of a
vindoline part attached to the catharanthine moiety via a
covalent bond.
VINCRISTINE
 Molecular Formula: C₄₆H₅₆N₄O₁₀
 IUPAC Name: (3aR,3a1R,8S,8aS,9R,9aS,9bS)-8-(formylmethyl)-9-hydroxy-
3a-(methoxymethyl)-3a1,5,8a,9a,9b-hexahydro-2H,6H-8,9-
(epimino)indolo[1,2-a]indol-6-one.
 Structure: Vincristine consists of a catharanthine and vindoline moiety. Its
distinguishing feature is the presence of an N-formyl group (-CHO) on the
vindoline part. This aldehyde group contributes to its interaction with
tubulin.
 Functional Groups:
• Indole ring: Core structure from catharanthine.
• Dihydroindole moiety: From vindoline.
• Formyl group: Attached to the vindoline, giving vincristine its distinctive
reactivity.
• Multiple hydroxyl groups (-OH): Involved in hydrogen bonding and
solubility.
VINBLASTINE
 Molecular Formula: C₄₆H₅₆N₄O₉
 IUPAC Name: (3aR,3a1R,8S,8aS,9R,9aS,9bS)-8-(formylmethyl)-9-hydroxy-
3a-(methoxymethyl)-3a1,5,8a,9a,9b-hexahydro-2H,6H-8,9-
(epimino)indolo[1,2-a]indol-6-one.
 Structure: Vinblastine shares the same basic framework as vincristine, but
without the formyl group on the vindoline moiety. Instead, it has a methyl
group (-CH₃). This slight structural difference affects the molecule's
pharmacodynamics, leading to different clinical applications.
 Functional Groups:
• Indole ring: From catharanthine.
• Dihydroindole moiety: From vindoline.
• Hydroxyl groups: On both moieties for solubility and interaction with
biological molecules.
STRUCTURE ACTIVITY
RELATIONSHIP (SAR)
 Catharanthine (Indole) Moiety:
This part of the molecule plays a key role in the interaction with tubulin.
Modifications in this region often reduce the drug's affinity for tubulin, thereby
decreasing its anti-mitotic activity.

 Vindoline (Dihydroindole) Moiety:

• This section contributes to drug solubility, metabolism, and side-effect
profile.
• The hydroxyl group in the vindoline moiety is essential for the cytotoxicity
of vinca alkaloids. Its removal significantly reduces the drug’s activity.
• Structural variations in the vindoline ring, such as methyl substitutions, can
influence both the potency and selectivity of the drug.
 Vincristine: Contains a formyl group (-CHO) at the vindoline moiety,
which contributes to its neurotoxicity.
 Vinblastine: Has a methyl group (-CH₃) instead of a formyl group, which
makes it less neurotoxic compared to vincristine but more
myelosuppressive.
 Vindesine: Contains an ethyl group (-C₂H₅) at the same position, resulting
in a balanced toxicity profile.
 Vinorelbine: Lacks a substitution at the same position, contributing to its
distinct pharmacological properties and improved therapeutic index in lung
cancers.
 The overall stereochemistry and rigidity of the structure are crucial for
binding to the tubulin β-subunit, which prevents microtubule
polymerization.
Subtle modifications to the molecule, such as stereochemical alterations, can
lead to loss of affinity for tubulin and a corresponding drop in anticancer
activity.
BIOSYNTHESIS
ISOLATION
 A. Collection and Preparation of Plant Material
• Source: The vinca alkaloids are isolated from Catharanthus roseus leaves
and stems. The plant can be harvested either fresh or dried.
• Drying and Grinding: The plant material is air-dried and then ground into
a fine powder. This increases the surface area and helps improve the
efficiency of the solvent extraction process.
 B. Extraction Process
• Solvent Extraction: The alkaloids are extracted from the powdered plant
material using polar organic solvents such as methanol, ethanol, or a
mixture of methanol and water.
• The dried plant powder is soaked in the solvent, and the solution is stirred
for several hours or days to ensure maximum extraction of the alkaloids.
• The solvent is then separated from the plant residue via filtration.
• Concentration: After extraction, the solvent is evaporated under reduced
pressure using a rotary evaporator to obtain a crude extract containing
alkaloids.
 C. Acid-Base Partitioning
• The crude extract is treated with an acid solution (typically dilute sulfuric or
hydrochloric acid), which converts the alkaloids into their salt forms. These
salts are water-soluble and can be extracted into the aqueous layer.
• The aqueous layer containing the alkaloid salts is separated from the
organic phase and then basified with a base (ammonium hydroxide or
sodium carbonate) to regenerate the free alkaloid bases, which can then be
extracted into an organic solvent (e.g., chloroform or ethyl acetate).
• Separation of Alkaloids: The resulting organic phase contains the
alkaloids, which are further concentrated by evaporating the solvent.
PURIFICATION
 A. Column Chromatography
• Step 1: The crude extract is subjected to column chromatography using a
stationary phase such as silica gel or alumina. The mobile phase can be an
organic solvent system like chloroform or methanol, which elutes alkaloids
at different rates based on their polarity.
• Step 2: Fractions are collected during the chromatography process, and
the alkaloid-containing fractions are pooled together.
 B. High-Performance Liquid Chromatography (HPLC)
• For higher purity, HPLC is used as a secondary purification technique.
• Alkaloids are separated based on their interaction with the stationary phase
in the column and the solvent gradient used as the mobile phase.
• The fractions collected from HPLC can achieve high purity levels and are
analyzed to determine which fractions contain the desired vinca alkaloid.
 C. Crystallization
• Crystallization is often employed to further purify the isolated alkaloids.
The alkaloids are dissolved in a suitable solvent and allowed to crystallize
out by slowly evaporating the solvent or cooling the solution.
• Recrystallization may be repeated multiple times to enhance purity.
CHARACTERIZATION
 A. Thin Layer Chromatography (TLC)
• TLC is used to monitor the purification process. The alkaloids can be
visualized using UV light or specific staining reagents.
• TLC provides a quick and effective way to check the progress of the
purification and confirm the presence of vinca alkaloids in the samples.

 B. Mass Spectrometry (MS)

• Mass spectrometry is used to determine the molecular weight and
structural fragments of the isolated alkaloids.
• This technique provides critical information on the exact molecular
composition of the alkaloid and helps in identifying specific vinca alkaloids
based on their mass spectra.
 C. Nuclear Magnetic Resonance (NMR) Spectroscopy
• 1H-NMR and 13C-NMR are used for detailed structural elucidation of the
alkaloids.
• NMR spectroscopy allows scientists to determine the positioning of
hydrogen and carbon atoms in the molecule, revealing the full structure
of the alkaloid.
• This technique is crucial for confirming the identity of the alkaloid and
distinguishing between different types of vinca alkaloids.

 D. Infrared (IR) Spectroscopy

• IR spectroscopy is used to identify functional groups present in the
alkaloid molecule, such as hydroxyl, amine, or methoxy groups.
• The absorption bands in the IR spectrum correspond to specific bonds
within the alkaloid, providing further structural insights.
 E. Ultraviolet-Visible (UV-Vis) Spectrophotometry
• UV-Vis spectrophotometry is used to analyze the absorbance
characteristics of the alkaloids in the UV-visible spectrum.
• This method is often used for the quantitative analysis of alkaloids in
pharmaceutical formulations, as it allows for the determination of
concentration based on absorbance.

 F. X-Ray Crystallography
• For the highest level of structural confirmation, X-ray crystallography can
be employed if high-quality crystals of the alkaloid are obtained.
• This technique provides a three-dimensional visualization of the alkaloid's
molecular structure, confirming bond lengths, angles, and overall
geometry.
UV SPECTRUM
 Vincristine:
• Absorption Peaks: Typically shows strong absorbance around 210 nm and
250 nm, which are characteristic of the aromatic rings and other functional
groups present in the structure.
• Solubility: It is often dissolved in solvents like methanol or DMSO for
spectral analysis.
 Vinblastine:
• Absorption Peaks: Similar to vincristine, vinblastine shows significant
absorbance around 220 nm and 250 nm. The specific peak positions may
vary slightly due to differences in their chemical structures.
• Solubility: Also commonly analyzed in methanol or other organic solvents.
IR SPECTRUM
In the IR spectrum of vincristine, we would typically observe:
 O-H Stretching: Broad peak around 3200-3500 cm⁻¹, indicative of alcohol
or phenolic groups.
 C-H Stretching: Peaks around 2800-3000 cm⁻¹ from aliphatic and
aromatic C-H bonds.
 C=O Stretching: Peaks around 1650-1750 cm⁻¹ if carbonyl groups are
present (though vincristine itself has limited carbonyls).
 C=C Stretching: Peaks around 1600-1680 cm⁻¹ related to aromatic rings.
 N-H Stretching: Peaks near 3300 cm⁻¹ due to the presence of amine
groups.
MASS SPECTRUM
 Vincristine
Molecular Weight: Approximately 824 g/mol.
Key Fragments:
The molecular ion (M⁺) peak, usually observed at m/z 824.
Fragments corresponding to the loss of various groups (e.g., loss of CH₃,
C₃H₄N) can appear, leading to smaller peaks at m/z values such as 807, 793,
and so on.
 Vinblastine
Molecular Weight: Approximately 811 g/mol.
Key Fragments:
The molecular ion (M⁺) peak at m/z 811.
Similar fragmentation patterns as vincristine, with peaks for the loss of CH₃
and other moieties.
13 C NMR SPECTRUM
H NMR SPECTRUM
Chemical Shifts:
 Aromatic protons typically appear in the range of 6.0 to 8.0 ppm.
 Aliphatic protons are generally found downfield (around 0.5 to 3.5 ppm).

400 MHz 1H NMR of standard

vinblastine sulphate
USES
 Cancer Treatment:
Vinca alkaloids, such as vincristine and vinblastine, are commonly used in
chemotherapy regimens for cancers like leukemia, lymphoma, and certain solid
tumors (e.g., breast and lung cancer).
 Vincristine
• Mechanism of Action: Vincristine binds to tubulin, preventing microtubule
formation, which disrupts the mitotic spindle during cell division, leading to
cell cycle arrest in the metaphase.
• Indications:
 Acute Lymphoblastic Leukemia (ALL): Often a key component of
combination chemotherapy regimens.
 Hodgkin’s Lymphoma: Used as part of multi-drug regimens.
 Neuroblastoma: Commonly used in pediatric cases.
 Wilms' Tumor: Used in some treatment protocols.
 Small Cell Lung Cancer: Sometimes included in combination therapy
 Vinblastine
• Mechanism of Action: Similar to vincristine, it disrupts microtubule
formation, leading to cell cycle arrest.
• Indications:
 Hodgkin’s Lymphoma: Integral to the ABVD regimen (Adriamycin,
Bleomycin, Vinblastine, Dacarbazine).
 Non-Hodgkin’s Lymphoma: Used in various combination therapies.
 Testicular Cancer: Utilized in treatment regimens for advanced cases.
 Kaposi’s Sarcoma: Sometimes used, especially in AIDS-related cases.
 Gestational Trophoblastic Neoplasia: Included in treatment protocols for
this condition.
 Research Applications
Vinca alkaloids are studied for their potential effects beyond oncology,
including:
• Autoimmune Disorders: Investigated for their immunosuppressive
properties.
• Neurobiology: Research on their effects on neural pathways and potential
neuroprotective roles.
DRUG INTERACTIONS
 1. CYP450 Interactions
Metabolism: Both vincristine and vinblastine are metabolized by the liver
enzyme CYP3A4. Drugs that inhibit or induce this enzyme can affect the levels
of vinca alkaloids.
• Inhibitors (e.g., ketoconazole, erythromycin): May increase vinca alkaloid
levels, leading to enhanced toxicity.
• Inducers (e.g., rifampin): May decrease vinca alkaloid levels, potentially
reducing their effectiveness.
 2. Corticosteroids
 Dexamethasone: Commonly used in conjunction with vinca alkaloids for
their antiemetic properties and to mitigate inflammation, but can also
affect metabolism and immune response.
 3. Antibiotics
 Quinolones (e.g., ciprofloxacin): May interact with the
pharmacokinetics of vinca alkaloids, although clinical significance is not
well defined.
 4. Grapefruit Juice
 Effect on Metabolism: Grapefruit juice can inhibit CYP3A4 and may
increase levels of vinca alkaloids, leading to heightened toxicity.
 5. Anticonvulsants
 Phenytoin: Can alter the metabolism of vinca alkaloids, leading to reduced
effectiveness or increased toxicity.
 6. Vaccines
 Live Vaccines: Patients receiving vinca alkaloids may have compromised
immune function, making live vaccines (e.g., MMR, varicella) potentially
dangerous.
 7. Digoxin
Vinca alkaloids may alter the pharmacokinetics of digoxin, potentially leading
to increased digoxin levels and toxicity.
SIDE EFFECTS
 Common Side Effects:
 Neurotoxicity: Peripheral neuropathy, characterized by numbness,
tingling, and weakness, is particularly common with vincristine.
 Constipation: Due to its effects on the gastrointestinal tract; preventive
measures may be necessary.
 Bone Marrow Suppression: Can lead to decreased blood cell counts,
increasing infection risk.

 Serious Side Effects:

 Extravasation Injury: If vincristine or vinblastine leaks into surrounding
tissues, it can cause significant local damage.
 Alopecia: Hair loss can occur, but is generally less severe compared to
other chemotherapy agents.
RECENT ADVANCEMENTS
 Nanoparticle Systems:
Research is ongoing into nanoparticle delivery systems that can encapsulate
vinca alkaloids, enhancing bioavailability and targeting capabilities, thus
reducing systemic toxicity.
 Microsphere Technology:
Development of microspheres to deliver vincristine in a controlled manner is
being explored to improve efficacy and safety.
 Targeting Specific Pathways:
Research is focusing on combining vinca alkaloids with agents that target
specific signaling pathways involved in cancer cell proliferation, such as the
PI3K/AKT/mTOR pathway.
 Non-Oncological Applications:
Trials are being conducted to explore their use in neurodegenerative diseases,
with potential applications in conditions like Alzheimer’s disease due to their
effects on microtubule stability.
MARKTED PRODUCTS
REFERENCES
 “Reviews on Indian Medicinal Plants”, Indian Council of Medical Research,
Volume 5, 2007, Page no.- 815-819.
 “Quality Standards of Indian Medicinal Plants”, Indian Council of Medical
Research, Volume 2, 2005, Page no.- 56-62.
 Kokate, C.K. (2017), “Textbook of Pharmacognosy”, Nirali Prakashan, Page
no. 15.28-15.30.
 Khare C.P. , Encyclopedia of Indian Medicinal Plants, Page no. 471-472.
 [Link]
 [Link]
 [Link]
THANK YOU !!