Title: Drug Discovery and Development Process of Anti-diabetic Plants
1Drug 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 bknanjwade_at_yahoo.co.in
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2Sources of drugs
Animal Insulin (Pig, cow)
Growth hormone (Man) (Creutzfeldt-Jakob) Plant
Metformin (Gallega officinalis )
Morphine (Papaver somniferum) Inorganic Arsenic,
Mercury, Lithium
Synthetic Chemical (Propranolol)
Biological (Penicillin)
Biotechnology (Human insulin)
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3Drug Discovery
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4Drug Discovery
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5The Regulatory process
- 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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6Marketing
- 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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7Schema diagram representing anti diabetic plants
data
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8A screen shot of the database Database on
anti-diabetic plants home page with links and
dropdown search window.
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9 Annona squamosa (SugarApple)
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10Nigella sativa
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11Polyherbal 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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12INTRODUCTION
-
- 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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13Animals
- Male Wistar albino rats (weighing 160200 g) were
procured from Venkateshwara Enterprise, Bangalore
and they kept in under standard environmental
conditions (12 h light/dark cycles at 2528 0C,
6080 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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14Preparation 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 1l of
water. This suspension was filtered and the
filtrates were pooled and the solvents were
evaporated in a rotavapor at 4050 0C under
reduced pressure and lyophilized.
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15Chemicals
- 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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16Drug 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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17Streptozotocin-induced diabetes
- Rats were made diabetic by single administration
of streptozotocin (60 mg/kg/i.p) dissolved in 0.1
M-citrate 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 gt 250 mg/dl) were used in
the experiment.
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18Experimental 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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19Experimental design
- 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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20Experimental 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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21Experimental 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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22Table 1 Polyherbal Formulation of Annona
Squamosa and Nigella Sativa (Composition and
Concentration).
Sl. No. Botanical Name Common Name Family Part used Conc. ()
1 Annona squamosa Sharifa Annonnaceae Matured fruits 50
2 Nigella sativa Kalonji Ranunculaceae Seeds 50
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23Table 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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24Pharmaceutical 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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25Building-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 products
performance - identification of drug products critical
characteristics (which would be monitored on a
batch-by-batch basis) - demonstration of drugs safety and efficacy
- ultimately leads to the review and approval of
the drug
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26Relationship 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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27Drug Development Process
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28Drug Development Process
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29Quality is Always Part of the Picture - Built-In
and Built-Up
Quality Control and Quality Assurance
Less established
Fully established
Commercial Manufacturing
Pre-IND
Phase I
Phase II
Phase III
Specification/Manufacturing Development for the
Product
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30Drug Development Cycle
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31Examples 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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32Chemistry 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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33Examples 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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34Process 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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35Validation
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36What 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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37Validation is Always Part of the Picture
Commercial Manufacturing
Pre-IND
Phase I
Phase II
Phase III
Final process validation
Specification Development
Re-validation
Ongoing Validation (DOE, IQ, OQ, PQ, PV)
DOE Design of Experiment IQ Installation
Qualification OQ Operational Qualification
PQ Performance Qualification PV Process
Validation
- The extent of IQ, OQ, PQ, validation, etc.
depends on complexity of product
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38Role 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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39Pre-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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40Foreign 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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41Foreign 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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42Chemistry 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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43Chemistry 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 to Quality
Control.
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44Applicability 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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45Studies 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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46Studies 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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47Natural 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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48Natural 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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49Conclusions
- 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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49
BIRDEM ANRAP, Dhaka, Bangladesh
50Cell No 0091 974243100 E-mail
bknanjwade_at_yahoo.co.in
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