Transcript Drug Discovery and Development Process of Anti

Drug Discovery and Development
Process of Anti-diabetic Plants
Dr. Basavaraj K. Nanjwade M. Pharm., Ph. D
Professor of Pharmaceutics
KLE University College of Pharmacy
BELGAUM- 590010, Karnatka, India.
Cell No: 0091 974243100
E-mail: [email protected]
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Sources of drugs
Animal
Insulin (Pig, cow)
Growth hormone (Man) (Creutzfeldt-Jakob)
Plant Metformin (Gallega officinalis )
Morphine (Papaver somniferum)
Inorganic Arsenic, Mercury, Lithium
Synthetic
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Chemical (Propranolol)
Biological (Penicillin)
Biotechnology (Human insulin)
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Drug Discovery
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Drug Discovery
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The Regulatory process
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Differs from country to country
Demands safety and quality of product
Encourages efficacy and need for product
Grants clinical trials certificate if volunteer and animal data OK
Approves protocols and examines data
50-400 volumes (30,000-150,000 pages)
Original data available
Two way process; authority and company trying to produce a safe
effective product
• Release for a specific purpose and use
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Marketing
• Getting the product right (packaging;
formulation)
• Right therapeutic slot
• Information on new drug
• Information for honest comparison
• Reporting problems
• Reporting new indications
• Therapeutic trends
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Schema diagram representing anti diabetic plants
data
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A screen shot of the database “Database on antidiabetic plants” home page with links and dropdown
search window.
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Annona squamosa (Sugar–Apple)
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Nigella sativa
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Polyherbal formulation of Annona
squamosa and Nigella sativa
This study was undertaken to investigate the effect of Polyherbal formulation
of Annona squamosa and Nigella sativa on blood glucose, plasma insulin,
tissue lipid profile, and lipidperoxidation in streptozotocin induced diabetic
rats. Aqueous extract of Polyherbal formulation of Annona squamosa and
Nigella sativa was administered orally (200 mg/kg body weight) for 30 days.
The different doses of Polyherbal formulation on blood glucose and plasma
insulin in diabetic rats were studied and the levels of lipid peroxides and
tissue lipids were also estimated in streptozotocin induced diabetic rats. The
effects were compared with tolbutamide. Treatment with Polyherbal
formulation and tolbutamide resulted in a significant reduction of blood
glucose and increase in plasma insulin. Polyherbal formulation also resulted
in a significant decrease in tissue lipids and lipid peroxide formation. The
decreased lipid peroxides and tissue lipids clearly showed the
antihyperlipidemic and antiperoxidative effect of Polyherbal formulation
apart from its antidiabetic effect.
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INTRODUCTION
Diabetes mellitus is syndrome, initially characterized by a loss of glucose
homeostasis resulting from defects in insulin secretion, insulin action both
resulting impaired metabolism of glucose and other energy yields fuels
such as lipids and protein. Experimental diabetes in animals has provided
considerable insight into the physiologic and biochemical derangement of
the diabetic state. Many of the derangement have been characterized in
hyperglycemic animals. Significant changes in lipid metabolism and
structure also occur in diabetes. In these cases the structural changes are
clearly oxidative in nature and are associated with development of vascular
disease in diabetes. In diabetic rats, increased lipidperoxidation was also
associated with hyperlipidemia. Liver, an insulin dependent tissue that
plays a pivotal role in glucose and lipid homeostasis and it is severely
affected during diabetes.
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Animals
• Male Wistar albino rats (weighing 160–200 g) were
procured from Venkateshwara Enterprise, Bangalore
and they kept in under standard environmental
conditions (12 h light/dark cycles at 25–28 0C, 60–
80% relative humidity) in clean and dry cages and
maintained in well-ventilated animal house. Animals
were divided into 8 groups of five each and were fed
with standard diet and water ad libitum. The study
was approved by the Institutional Animal Ethics
Committee.
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Preparation of drug
• The seeds of Nigella sativa obtained from Prgati Ayurvedic
Drug store Belgaum and matured fruit of Annona squamosa
from local market of Belgaum and they were authenticated
from Botanical Survey of India, Pune (Maharastra).
• The extracts of the both antidiabetic plants were mixed and
polyherbal formuation was prepared (Table 1). Five hundred
grams of each plant (chopped into small pieces) was extracted
individually and were soaked overnight in 1:l of water. This
suspension was filtered and the filtrates were pooled and the
solvents were evaporated in a rotavapor at 40–50 0C under
reduced pressure and lyophilized.
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Chemicals
• Streptozotocin and other biochemicals used in this
experiment were purchased from Sigma Chemical
Company Inc., St. Louis, Mo, and USA. Enzyme
linked immunosorbant assay (ELISA) kit for insulin
assay was purchased from Boehringer Mannheim,
Germany.
• Tolbutamide was a generous gift sample from Sun
Pharmaceuticals Limited, Baroda, India. All other
chemicals used were of analytical grade.
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Drug administration
• Polyherbal formulation of Annona squamosa
and Nigella sativa was suspended in distilled
water and administered orally through
intragastric tube at the following doses of 50,
100 and 200 mg/kg body weight.
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Streptozotocin-induced diabetes
• Rats were made diabetic by single administration of
streptozotocin (60 mg/kg/i.p) dissolved in 0.1 Mcitrate buffer, pH 4.5. Forty-eight hours later, blood
samples were collected and glucose levels were
determined to confirm the development of diabetes.
• Only those animals which showed hyperglycemia
(blood glucose levels > 250 mg/dl) were used in the
experiment.
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Experimental design
• In the experiment, a total of 42 rats (30
diabetic surviving rats, 12 normal rats) were
used. The rats were divided into seven groups
of six rats each after the induction of
streptozotocin diabetes.
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Experimental design
1. Group1: Normal treated rats.
2. Group2: Normal rats given aqueous solution of Polyherbal
formulation (200 mg/kg body weight) daily using an
intragastric tube for 30 days.
3. Group 3: Diabetic control rats.
4. Group 4: Diabetic rats given aqueous solution of Polyherbal
formulation (50 mg/kg body weight) daily using an intragastric
tube for 30 days.
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Experimental design
5. Group 5: Diabetic rats given aqueous solution of Polyherbal
formulation (100 mg/kg body weight) daily using an
intragastric tube for 30 days.
6. Group 6: Diabetic rats given aqueous solution of Polyherbal
formulation (200 mg /kg body weight) daily using an
intragastric tube for 30 days.
7. Group 7: Diabetic rats given aqueous solution of Tolbutamide
(250 μg/kg bodyweight) daily use an intragastric tube for 30
days.
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Experimental design
• At the end of 30 days, all the rats were killed by
decapitation under pentobarbitone sodium (60 mg/kg)
anesthesia. Blood was collected in tubes containing
potassium oxalate and sodium fluoride solution for
the estimation of blood glucose and plasma was
separated for the assay of insulin.
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Table 1: Polyherbal Formulation of Annona
Squamosa and Nigella Sativa (Composition and
Concentration).
Sl. No.
Botanical
Name
Common
Name
Family
Part used
Conc.
(%)
50
1
Annona
squamosa
Sharifa
Annonnaceae
Matured
fruits
50
2
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Nigella
sativa
Kalonji
Ranunculaceae Seeds
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Table 2: Changes in blood glucose and plasma
insulin levels of control and experimental animals
Group
Fasting blood glucose
(mg/dl)
Plasma insulin (IU/ml)
Normal
81.04 ± 2.29
11.26 ± 0.96
Diabetic control
262.24 ± 22.23
3.48 ± 0.69
Diabetic + Polyherbal
formulation (50 mg/kg)
209.58 ± 12.46
5.59 ± 0.34
Diabetic + Polyherbal
formulation (100 mg/kg)
155.58 ± 11.69
6.03 ± 0.45
Diabetic + Polyherbal
formulation (200 mg/kg)
104.16 ± 6.56
7.15 ± 0.45
Diabetic + Tolbutamide
(250 mg/kg)
110.65 ± 9.35
6.32 ± 0.48
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Pharmaceutical Product Quality
Cannot Be Tested in - It Is Built in
• Pharmaceutical product quality is assured by
– comprehensive development program
– extensive manufacturing and environmental controls
– rigorous validation procedures and requirements
• The high quality thus built into the final product is
ensured through in-process controls and verified
in a series of confirmatory tests before each
manufactured batch is released to the market
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Building-in of Quality Starts Early.
Development Builds-in Quality
• The chemistry, manufacturing and controls (CMC)
aspect of drug development is focused on producing
medicines suitable for human use with specified
quality, safety and efficacy characteristics
• The drug development program is geared towards
– thorough understanding of the drug product’s performance
– identification of drug product’s critical characteristics
(which would be monitored on a batch-by-batch basis)
– demonstration of drug’s safety and efficacy
– ultimately leads to the review and approval of the drug
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Relationship between
Safety, Efficacy and Quality
• Every drug product (with its specifications) has
been thoroughly tested in clinical trials for
safety and efficacy
– Specifications for release and stability testing may
be equal to or tighter than the specifications for
clinical trial batches
• Therapeutic indication and QC are considerations in
establishing specifications
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Drug Development Process
% of all
new drugs
that pass
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Phase II
Phase III
FDA
Approval
3.5
1-2
2-4
4-6
1.5
Total = 12 - 17
Laboratory
and Animal
Studies
20 to 100 Healthy
Volunteers
100 – 300 Patient
Volunteers
1,000 to 3,000
Patient Volunteers
Assess
Safety and
Biological
Activity
FILE IND
Phase I
Determine Safety
and Dosage
Evaluate
Effectiveness. Look
for Side Effects.
Verify Effectiveness,
Monitor Adverse
Reactions from LongTerm Use
70% of INDs
30% of INDs
27% of INDs
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Post Marketing
Safety Monitoring
FILE NDA
Purpose
Test Population
Years
Pre Clinical
Testing
Review
Process
Large Scale
Manufacturing
-------------Distribution
-------------Education
20% of INDs
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Drug Development Process
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Quality is Always Part of the Picture Built-In and Built-Up
Quality Control and Quality Assurance
Less established
Pre-IND
Fully established
Phase I
Phase II
Phase III
Commercial
Manufacturing
Specification/Manufacturing Development
for the Product
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Drug Development Cycle
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Examples of QA & QC Considerations
During Drug Development
• Evolution of documentation systems
– SOP
– change control
– trend analysis
• Evolution of QA and QC systems
– internal audits
– supplier audits
– document review (e.g., SOP, batch records, specifications,
data)
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Chemistry Manufacturing Controls
Evolve During Drug Development
• The goal is to have process and product
performance determined by the time of
validation, although some level of validation
occurs along the continuum and eventually
leads to the full-scale validation.
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Examples of CMC Considerations
During Drug Development
• Selection of appropriate technology and raw materials
• Optimization
– of formulation and device
– of manufacturing process
– of specifications and analytical methods
• Careful selection and control of container closure systems
• Identification and control of critical manufacturing process
parameters
• Process capability established
• Technical transfer to larger scale, i.e., scale-up
• Process validation
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Process Registration Requirements
• Sponsor is required to describe how the product was developed
• Companies need to optimize, justify and register the entire “recipe”
– ranges
• temperatures
• mixing times
• hold times
• etc.
– quantities
• active ingredient
• excipients
– raw material specifications
– in-process limits
– in-process methods
– product specifications
– etc.
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Validation
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What is Validation?
• Documented evidence that the manufacturing process
consistently produces product that meets predetermined
specifications
– Defines product quality
– Developed and validated based on a thorough
understanding of the critical process parameters
– Parameters are carefully controlled within the validated
ranges to ensure a consistent manufacturing process.
• Manufacturing process validation consists of successfully
manufacturing at least three full-scale batches in
succession, which pass all in-process and product quality
attributes
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Validation is Always Part of the Picture
Pre-IND
Phase I
Phase II
Phase III
Commercial
Manufacturing
Final process validation
Specification Development
Re-validation
Ongoing Validation
(DOE, IQ, OQ, PQ, PV)*
• The extent of IQ, OQ, PQ, validation, etc.
depends on complexity of product
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* DOE = Design of Experiment
IQ = Installation Qualification
OQ = Operational Qualification
PQ = Performance Qualification
PV = Process Validation
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Role of QC Tests
• Each batch of orally inhaled and nasal drug products
(OINDP), manufactured by the validated process, is
tested to the critical QC attributes as defined during
development
– Confirms consistent performance
• The Delivered Dose Uniformity test for OINDP is one of
several confirmatory QC tests of the finished product
– a result of a long and careful development and characterization
process
– QC tests confirm the quality built-in through a well-understood
and well-controlled manufacturing process
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Pre-Approval Inspection
• Confirms Facility is Ready
– Sponsor can do what they submit in the NDA
– Process is validated or validation protocols are in
place
• Validation required prior to launch
– Thorough documentation review
– Quality systems are established and capable
– Confirms specifications are met
• Compliance versus Review Division
– Specifications may change based on NDA review
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Foreign Marketing Data
• Several Ayurvedic, Siddha and Unani drugs have been
marketed for centuries in India. The drugs are also dispensed
to the patients by recognized and qualified physicians of the
complimentary systems.
• Most of these drugs have been found to be safe, It may be
worthwhile for the, FDA to accord adequate weightage to the
data already available in these countries in determining
whether a “drug has been used under particular conditions to a
material extent and for a material time” to qualify for inclusion
in an OTC drug monograph.
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Foreign Market Data
• Since the FDA is proposing to make it
available the facility for OTC marketing
agencies in the United States, who are already
marketing their products, the same exemptions
can be made available to overseas marketing
firms who have safely-marketed their products
in their respective countries.
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Chemistry Manufacturing Controls (CMC)
information for the botanical drug products
• Botanical, drugs. are derived from vegetable matter
and are usually complex mixtures. The chemical
constituents of such a mixture are not, always defined
and in most cases even the active constituent in a
botanical drug is not defined nor its biological
activity well characterized.
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Chemistry Manufacturing Controls (CMC)
information for the botanical drug products
• Therefore the CMC documentation would have to be
substantially liberal in comparison to that’ of
synthetic or highly purified drugs. Simple or
combination
tests
like
spectroscopic,
chromatographic, fingerprints,, chemical and or
biological assays can be the main reliability criteria to
understand the product.
• While these tests may not generate the necessary
specifics that are desirable, it can atleast bring about a
rational approach ANRAP
to Quality
Control.
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Applicability of Combination
Drug Regulations
• It would be practical not to confine the botanical drug
products that are derived from a single part of a plant
such as leaves, stems, roots seeds under the fixed
combination drug category.
• The current requirement of Botanical drugs composed
of multiple parts of a single plant species under the
combination requirement should also be revised and
exemption accorded.
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Studies on bioavailability and
drug interactions
• Bioavailability and Pharmacokinetic studies are
cumbersome and extremely difficult to generate in
complex formulations, where a number of herbs are
involved. It would be desirable to have” a very
practical view in this area.
• Well-controlled clinical trials can substitute the
bioavailability and Pharmacokinetic studies.
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Studies on bioavailability and
drug interactions
• Ayurvedic and Siddha pharmacopoeia are full of
formulations which have a. combination of several
herbs and most of them can never be launched in the
next decade or two if requirements are not simplified
by the FDA.
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Natural Vs Cultivable source of
Plants
• In a Country like India where the source of raw
materials for Drug preparations is predominantly
from the forests (80%) a practical solution has to be
found in connection with the source of the raw
material for quality enforcements.
• When a medicinal plant is cultivated a lot of the
Quality aspects are directly under the control of the
grower.
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Natural Vs Cultivable source of
Plants
• However, when the plant material is collected from
the forests, it would be reasonable to presume that the
origin of the source is, from nature and variations if
any would be natural.
• The document should highlight separate guidelines
for natural and cultivable sources for manufacturers
using plants from the above two sources
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Conclusions
• Pharmaceutical quality is built-in through the
entire drug development process
– validation is key element of ensuring quality
– in-process controls assure quality during
manufacturing
– Specifications established based on thorough
understanding of process
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THANK YOU
Cell No: 0091 974243100
E-mail: [email protected]
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