Product Development and
Clinical Studies of
Traditional Medicines
N. L. Phang
Nova Laboratories Sdn. Bhd.
Malaysia
(www.nova.com.my)
Objectives
1. To discuss the main issues and regulatory guidelines on product development.
2. To describe develop process of a traditional medicine with therapeutic claim.
To illustrate with a case study–
The development of a botanical drug containing standardized Phyllanthus niruri extract EPN 797.
Successful herbal product:Development of bromhexine
Ethnobotany approach in drug development
• Bromhexine (Trade name: Bisolvon) – Is a popular mucolytic agent for cough.
– It is derived from Indian Adhatoda vasica (the malabar nut tree) which is traditionally used in cough therapy.
Definition of herbal products
Three categories of herbal products:
A. Drugs (NCE, new chemical entities)
Single active ingredient pharmaceuticals originating from plants.
E.g.: vinblastine, digoxin.
Definition of herbal products (Cont’d)
B. Botanical drugs (Multi-component botanical drugs)Botanical drugs are manufactured medicines obtained exclusively from plants to relieve, prevent or cure a disease or to alter physiological and pathological process.E.g.: None in USA, several in clinical trials.
C. Dietary supplements / herbal supplementsPlant components with health benefits.E.g.: Garlic or Echinacea
Why do we develop botanical drugs?
1. Diverge chemical rangeEnormous propensity to synthesize diverse bioactive compounds.
(Multiple and mutually potentiating therapeutic effects)
Complex molecules with unique properties.
2. Biomanufacturing factories –
Relatively low-cost, highly effective and complex.
3. Higher investment cost on chemical synthesis.
4. Perception that phytochemicals provide a safer and more holistic approach to disease treatment and prevention.
Development process of botanical drugs
i. Development of botanical drugs is a hard and expensive task.
ii. Each new drug requires a big investment and a minimum of 5
- 10 years of work.
iii. Therefore, we have to adopt a very carefully planned strategy.
iv. The development of botanical drug emphasizes on three
important aspects of the product:
– Quality
– Efficacy
– Safety
– Inter-related – as efficacy and safety largely depend on the quality.
Herbal product development:The 6S Process *
Selection• Herb informatics• Unmet health need
Sourcing• Chemotaxonomy• Raw material analysis
Structure• Identification of active
constituents• Method validation
Standardization• Chemical profile• Pharmacological profile
Safety• Historical / traditional use• Toxin analysis• Safety studies in animals
Substantiation . Review of pre-existing data• Prospective clinical study
* Joseph Chang, Biochemical Pharmacology, Vol. 59, pp 211-219, 2000.
Study the guidelines for botanical drugs
• The final products are strictly controlled by the regulatory authorities.
• It is important to study the requirements of several important regulatory guidelines.
USA FDA CDER Guidelines for botanical drugs
• The Guidance for Industry: Botanical Drug Products – quality standards for standardized plant extracts (botanical drugs).
• Guidelines for the development of drug products from botanicals.
• It is now possible to market these products under the New Drug Application (NDA) approval process.
Abbreviated preclinical and clinical testing protocols
For plants with safe history of human use
USA FDA proposed abbreviated preclinical and
clinical testing protocols for botanical drugs
derived from plants.
Companies in North America and the UK currently involved in development of botanical drugs for clinical trials
Company Areas of clinical testing
Ancile pharmaceuticals. San Diego, CA Sleep, anxiety disorders
CV Technologies. Edmonton, Canada Respiratory infection
GW Pharmaceuticals. Salisbury, UK Cannabis-based prescription medicines
Oxford Natural Products. Oxford, UKDysmenorrhoea, hepatitis-C symptoms, cognitive decline
PhytoCeutica. New Haven, CT Cancer, neurovascular disease
Phytomedics. Dayton, NJ Autoimmune diseases, cancer
Phytopharm. Godmanchester, UKAppetite suppressant, inflammatory bowel disease, Alzheimer’s disease, cancer
WHO & EMEA guidelines
• The WHO Guidelines for the Assessment
of Herbal Remedies, adopted by the
International Conference of Drug
Regulatory Authorities (Ottawa, October
1991).
• EMEA Guidelines.
Pharmacopoeial monographs: Quality specification
• Define the quality standards of herbal
products.
• E.g: USP NF (USA), Indian Pharmacopoeia,
Chinese Pharmacopoeia.
Structure of USP botanical monograph Monograph for Gingko (USP26 NF21):
i. Definition of herbal product iv. Microbial limits
ii. Identification tests v. Limit tests – For soil and sand contamination
iii. Content tests – Quantitative determination of marker compounds
a. Content of flavonol glycosides by HPLC
b. Content of terpene lactones by HPLC
vi. Heavy metals
vii.. Pesticide residues
Indian Herbal Pharmacopoeia Volume 2: Monograph for Phyllanthus
Describes analytical methods (HPLC) of marker compounds: • Phyllanthin and Hypophyllanthin
hypophyllanthin
phyllanthin hypophyllanthin
phyllanthin
(a) Reference standards (b) Sample preparation
Chinese Pharmacopoeia Volume 1:Monograph for Ginkgo
-- Describes assay method (HPLC) for flavonol glycosides.
Main specification requirement of botanical drugs
Chemical standardization:
i. quality identification of the product.
ii. quantitative determination of the marker
compound(s) of the product.
Chemical standardization
Chemical standardization emphasizes the
importance of determination of the content
of the herbal products.
Importance of measurement
A quotation from Lord Kelvin:
“When you can measure what you are
speaking about, and express it in numbers,
you know something about it …”
Malaysian guidelines
-- Published by National Committee For Research And
Development In Herbal Medicine (NRDHM)
Guidelines for Standardization of
Herbal products
Guidelines for Levels and Kinds of Evidence to Support Claims for Therapeutic Products
-- Similar to FDA guidelines, they allow the development of
botanical drugs with therapeutic claim.
HEPAR-P capsule:Approved for clinical trial
• HEPAR-P Capsule contains standardized
Phyllanthus niruri extract.
• 1st local product approved by Medical
Research & Ethics Committee, Ministry of
Health Malaysia.
• For clinical testing to evaluate antiviral
activities in patients with chronic hepatitis B.
Two phases of development process:
Pre-clinical and clinical studies
1. Pre-clinical studies (Animal studies)
• Evaluate the pharmacological activities.
• Evaluate the toxicity.
2. Clinical studies (Human studies)
• Evaluate the efficacy.
• Evaluate the toxicity.
Flow chart of development of HEPAR-P Capsule (1)
Medical plant
Extract
Fractions ToxicologyBioassays
Chemical characterization
Flow chart of development of HEPAR-P Capsule (2 - Cont’d)
Standardized extract – EPN 797
Drug Delivery Technology
Manufacturing technology for
Finished product
Stability studies
Pure compound
New chemical entity
Chemical standardization
Qualitative & quantitative analytical methods
Flow chart of development of HEPAR-P Capsule (3 - Cont’d)
Complete monograph (of herbal product)
Approved herbal supplement
Animal toxicology studies (on the finished product)
-- Rodents & Non-rodents
-- According to FDA / WHO / Malaysian guidelines
Quality control protocol• Chemical standardization• Biological standardization
Flow chart of development of HEPAR-P Capsule (4 - Cont’d)
Completion of pre-clinical studies
Submission to National Committee for Research and Development In Herbal Medicine (NRDHM)
Approval to conduct clinical trial at appointed hospitals
Report of clinical trial by clinical investigators Recommendation by NRDHM
Application to DCA for registration with therapeutic claim
• LD 50• Acute toxicology studies • Sub-acute toxicology studies• Chronic toxicology studies
Specification of a botanical drug
i. Chemical standardization• Identification of chemical constituents• Measurement of marker compounds
ii. Biological standardization• In vitro anti-HBsAg activity (ELISA method)• In vivo liver protective activity in rats
iii. Stability of the finished product• Accelerated stability study• Real time stability study
Chemical structure:Corilagin & Phyllanthus flavonoids
Corilagin – Polyphenol
(Anti-viral & liver protective)
Phyllanthus flavonoids
(Liver protective)
Chemical structure of rutin, one of the Phyllanthus total flavonoids.Chemical structure of corilagin.
TLC fingerprint
Niranthin
HypophyllanthinPhyllanthin
TLC identification of Phyllanthin and fingerprints of HEPAR-P capsule
TLC fingerprints (Cont’d)
TLC identification of rutin in HEPAR-P capsule
Corilagin
TLC identification of corilagin in HEPAR-P capsule
Rutin
Chemical standardization:HEPAR-P Capsule
Content of one HEPAR-P Capsule
250 mg of Phyllanthus niruri extract EPN 797
standardized to contain:i. Corilagin 10 mgii. Total flavonoids 45 mg
Biological standardization
• Chemical standardization is inadequate.
• Botanical drug contains a complex mixture of chemical compounds.
• Chemical standardization does not give a complete picture of a herbal product.
• We have combined biological assays with chemical fingerprints to provide assurance of efficacy and consistency.
HEPAR-P Capsule:Quality control
• Chemical standardization
– Ensures batch-to-batch consistency in
chemical composition.
• Biological standardization
– Ensures batch-to-batch reproducible
biological activities.
Biological standardization
• Liver protective – In vivo (animal study)
• Anti-viral – In vitro (ELISA test)
Result of liver protective study
0
50
100
150
200
Control CCL4 control Treatment withPhyllanthus
0
50
100
150
200
Control CCL4 control Treatment withPhyllanthus
ALT
(U
/L)
AS
T
(U/L
)
Effect of Phyllanthus on CCL4 induced Liver injury in rats.
Effect of Phyllanthus on CCL4 induced Liver injury in rats.
Result of Inactivation of HBsAg study
In vitro inhibitory activity against
Hepatitis B surface antigen (HBsAg) by HEPAR-P™
HEPAR-P Capsule:Monograph
Quality specification (as compared to USP , IHP , CP)
• Definition of product
• Chemical identification
• Chemical assays for characteristic marker compounds
• Assays for biological activity (or biological assay)
• Heavy metals
• Microbial limits
Specification of HEPAR-P Capsule
Specification
Appearance description
Identity Impurities Potency Contaminants Quality
Color
SmellProduct related
Process related
Heavy metals
Microbial
Pesticide residues
Safety of HEPAR-P Capsule:Animal toxicology studies
Toxicology evaluation on the finished product:
i. Acute studies (14 days, rodents and non-rodents)
ii. Sub-acute studies (90 days , rodents and non-rodents)
iii. Chronic study – Rodents (180 days)
iv. Chronic study – Non-rodents (270 days)
Pre-clinical studies for HEPAR-P Capsule (Animal studies)
• Identification – The active fraction.• Characterization – The marker compound(s).• Establishment – Chemical standardization methods.
(Optional – Biological standardization methods)• Animal toxicology studies.• Stability studies.• Formulation development.• To establish a complete monograph of the product.• Medical Research & Ethnics Committee – approval to
conduct clinical trial.
Completion of pre-clinical studies:Approval for clinical trial
-- Approval by National Committee For
Research And Development In Herbal
Medicine (NRDHM).
-- Clinical trial at Selayang General
Hospital on Jan / Feb., 2005.
Clinical studies (Human studies)
• Clinical evaluation of safety and efficacy in human
subjects by appointed clinical investigators.
• Report of the clinical evaluation by the investigators.
• Recommendation by National Committee for
Research and Development in Herbal Medicine.
• Submission to DCA for registration of product as
botanical drugs with approved therapeutic claim.
Conclusion
1. The development of botanical drugs is likely to be a major area of plant biotechnology expansion in the 21st century.
2. The future of botanical drugs will depend on consumer and regulatory acceptance.
3. The important challenge is to provide science-based evidence to
consumers and regulatory authority on:
i. Efficacy (By clinical evaluation in human),
ii. Quality (Establish monograph of the standardized extract), and
iii. Safety (By toxicological evaluations in animals and in human).