DRUG DISCOVERY

1
 Plants as source of drugs
• Individual plant species often contain over
1,000 unique chemical entities (or
enzymatic machinery needed to produce
compounds upon the proper stimuli)
• Plants have to develop an extraordinary ary
of defenses, most of them chemical, to
protect themselves from viral diseases,
fungal pathogens, insects and mammalian
predators
2
• It is estimated that there are roughly
500,000 higher flowering plant species on
land
• A large number of these species has only
been very superficially examined for their
pharmacological and medical application
• Less than 1% of these species has been
thoroughly investigated for their potential
as therapeutic agents

3
 Evolution in drugs from plants
• The earliest drugs from plants took the form of
crude drugs such as tinctures, teas, poultices,
powders and other herbal formulations
• Later on drug discovery from medicinal plants
led to the isolation of early drugs like cocaine,
codeine, digitoxin, quinine and morphine
• More recently drug discovery techniques have
been applied to standardization of herbal
medicines, to elucidate analytical marker
compounds

4
 Goals of using plants as sources of
therapeutic agents are
• To isolate bioactive compounds for direct use
as drugs e.g. digoxin, digitoxin, morphine,
reserpine, taxol, vinblastine, vinceristine
• To produce bioactive compounds of novel or
known structures as lead compounds for semi-
synthesis to produce patentable entities of
higher activity and/or lower toxicity e.g.
metformin, nabilone, oxycodon, taxotere,
teniposide, verapamil and amiodarone which
are based, repectively on galegine, delta nine
tetrahydocannabinol, morphine, taxol,
podophyotoxin, khelline
5
 Cont.
• To use agents as pharmacologic tools, e.g. lysergic
acid diethylamide, mescaline, yohimbine
• To use the whole plant or part of it as a herbal
remedy e.g. cranberry, echnacea, feverfew, garlic,
ginkgo biloba and St. John’s wort
• Ethnomedicine (ethnobotanic medicine) is the use
of plants by humans as medicines
• Ethno-pharmacology is highly diversified approach
to drug discovery involving the observation,
description and experimental investigation of
indigenous drugs and their biologic activities

6
Importance of medicinal plants in
drug discovery
• Numerous methods have been utilized to
acquire compounds for drug discovery
including
– Isolation from plants and other natural sources
– Synthetic chemistry
– Combinatorial chemistry
– Molecular modeling
• Despite all that natural products and
particularly medicinal plants remain an
important source of new drugs, leads and new
chemical entities
7
Contribution of medicinal plants
to modern medicine
• Over 120 pharmaceutical products currently
in use are plant derived
• 75% of these were discovered by examining
the use of these plants in traditional
medicine
• A large proportion has come from tropical
forest species.

8
 Traditional medicine
• Is a comprehensive phrase which refers to
several traditional medical systems, such as
Chinese, African, Indian and other forms of
indigenous medical practices.
• In countries where the dominant health care
system is based on modern Western medicine,
or where traditional medicine has not been
incorporated into the national health system,
traditional medicine is often termed as
complementary, alternative or non-
conventional medicine.
9
 Traditional medicine, WHO definition:
• Traditional medicine is the sum of all
knowledge and practices, whether
applicable or not, used in diagnosing,
preventing or eliminating physical, mental
or social diseases and which may rely
exclusively on past experience and
observation handed down from generation
to generation, verbally or in writing.

10
• Worldwide, traditional or complementary medicine
is used to treat chronic pain and to improve the
quality of life for those suffering from incurable
diseases.
• Population depending on traditional medicine
worldwide:
– India for example 65% of population
– Asia and Latin America
– 48% in Australia
– 50% in Canada
– 42% in U.S.A
– 40% in Belgium
– 75% in France
11
 Traditional Healer
• A person recognized by the community in
which he lives as competent to provide
health care by using vegetable, animal and
mineral substances and certain other
methods based on the social, cultural and
religious background as well as on the
knowledge, attitudes and beliefs that are
prevalent in the community regarding
physical, mental and social well-being and
the causation of disease and disability.
12
 Choice of disease for study
• Should be guided by national epidemiological
data on the incidence and severity of the
disease
• The issue of availability, affordability,
effectiveness, safety and accessibility of
conventional medicines
• WHO (Africa) has identified five priority
diseases: maralia, HIV/AIDS, TB, Sickle-cell
disorder, hypertension and diabetes to be
initially targeted for research in and
development of African traditional medicine
13
 Methods in drug discovery
• Two main methods can be used:
– Ethno-medically driven approach
– Mass screening approach

14
Figure 1

A Comparison of Shaman's Ethnomedically Driven Drug Discovery Process with the Traditional Mass Screening Drug Discovery Approach.

      

15
16
 Ethno-medically driven drug
discovery process
• Random collection of plants for screening with
a variety of mechanism based assays with
specific applications to numerous therapeutic
areas (only successful pharmaceutical firms)
• A highly-focused collective program for
medicinal plants with a primary emphasis on
the use of plants by indigenous people in the
tropical regions of the world.
– Its advantage: it begins and ends with humans.

17
 How this method works
• Before any research expedition is undertaken:
– Prepare a full region study on the epidemiology,
traditional medicine, culture and ecology of the
people and the environment in which they live.
– Information of the plants known to be utilized in
any given area (use of data bases, medicinal
activities of plants and any known chemistry of
plants with such activity)
– Search for data in from international and national
hospitals in remote areas and treatment programs
that work with native and local people.

18
 After that
• Research teams are then sent to select tropical
regions to collect indigenous information and
witness first hand the use of medicinal plant
medicines to treat various illnesses.
– These field research teams are comprised of ethno-
botanists, medical doctors, local botanical
collaborators and indigenous healers and herbalists.
• This combination of expertise allows for a
highly focused selection of plant candidates for
screening and subsequent development.

19
 Advantage of the method
• Since the collected plants have a history of ethno-medical use in
humans, the need to screen thousands of plant species and
extracts is unnecessary
• In vivo laboratory screening serves as a confirmatory screen
• The most promising plant leads are then subjected to
fractionation campaigns
• Chemists isolate the entity responsible for activity
• Structure elucidation of the active compound
• Use the isolated natural compound as a template for further
structural modification to reduce toxicity and/or improve
potency
• As a result of this process new chemical leads can be generated
from the initial orally active natural product lead.

20
 Another approach
• An approach based on ethnomedical information to
experimentally pursue plants as a source of drugs
• It is designed primarily for implementation by
developing countries where lack of hard currency
often prevents sophisticated types of research from
being conducted.
• It is used for drug development in the form of
stable, standardized crude extracts and eventual
development of the active principles from these
plants
21
Deri v e nam e from Rev i ew E v al uate data i nDec i de on Determ i ne s afety
c atal ogue of frequl i terature Need to tes t from publ i s hed
l i terature s tudi es
entl y us ed pl ants

S el ec t tes ts

E s tabl i s h pri or-

ti es for tes ti ng Col l ec t pl ants

Col l ec t pl ants

Determ i ne Is ol ate and i de

Dev el op c ri teri a forP repare bi oas s ay pro Carry out tes ts ty pe of bi ol ogify ac ti v e pri nc
s afety and tox i c i ty toc ol s for s afety and c al ac ti v i ty pl es
tes ts tox i c i ty for s afety and
tox i c i ty
P repare, s tabi Carry
li out
z e and s tandar-hum an s tudi e
di z e ex trac ts

Dev el op m eth
of i ndus tri al p
duc ti on

22
 Some examples of drugs from plants that
served as models for next generation of drugs
• Khellin from Ammi visnaga (L.) Lamak. Was used as a
bronchodilator in U.S until it was shown to produce nausea and
vomiting after prolonged use.
• In England a group of chemists set about to synthesize khellin
analouges as potential bronchodilators with few side effects.
• This led to the discovery of cromoglycate as a sodium salt
(Intal) which stabilized cell membranes in the lungs to prevent
the allergen-induced release of the substance ultimately causing
brocho-constriction in allergic asthma patients
• Further studies elsewhere led to the synthesis of amiodarone
(Cordarone X) a useful antiarrythmia agent

23
 O OCH3

CH3 O O

OCH3
Khellin
|
O
O OCH2 CHCH2 O
O C R
OH
|
( C H2 ) 3 CH3
Na O2 C O O CO2 Na
R= OCH2 CH2 N( C H2 CH3 ) 2

Sodium cromoglycate Amioda rone

24
 Cont..
• Galegine was isolated as an active anti-
hyperglycemic agent from the plant Galega
officinalis L. which was ethnomedically
used for the treatment of diabetes. Galegine
provided the template for the synthesis of
metformin.
• It opened up interest in the synthesis of
other biguanidine-type antidiabetic drugs

25
CH3 O NH ( CH3 ) 2 N NH
C CHCH2 NHC C NHC
CH3 O NH2 NH NH2

Galengine Metformin

26
 Cont..
• Papaverine, useful as a smooth muscle
relaxant, provided the basic structure for
verapamil, a drug used to treat hypertension

27
CH3 O CH3 O

N N CH3 OCH3
CH3 O CH3 O
CH2 CH2 CH2 C OCH3
OCH3
( C H3 ) 2 HC CN
Papaverine
OCH3 Verapamil

S tructure re la tionship of ve ra pa mil to pa pa ve rine

28
 Advantages and disadvantages of using plants as
starting point in any drug development program
• Advantages:
• If one selects to use ethno medicine (long-term use by humans):
– Any isolated active compounds are likely to be safer than active
compounds with no history of human use.
• Plants are renewable source of starting materials in many cases
• Plants provide an unlimited source of novel and complex
chemical structures that most likely would not be the subject of
beginning synthetic programs.
• If the active principles derived from plants have novel structures
and useful biologic activity, patent protection can be assured
• The trend today in industry is to seek bioactive compounds from
plants that will serve as lead compounds for synthetic or semis-
synthetic development to assure patent protection

29
• Disadvantages: from random selection or
ethno medical claims
• Plants as biological systems have inherent
potential variability in their chemistry and
resulting biological activity
– Some plants showing promising biological
activity in assay systems fail to confirm the
activity in subsequent re-collection. This could
be due to:
• Variability in chemistry of plants or in the bioassay
systems used or mix-ups in labeling plants or
taxonomic identification

30
• The regulations of the Convention on Biological
diversity in 1992.
– Countries with the most biologic diversity have either
prohibited collection of plant material for export or
promulgated regulations that make it difficult to collect plant
samples.
– Several issues are tied in with the restrictions set forth by
countries including:
• preservation of their generic material.
• Intellectual property rights
• Compensation for discoveries arising from their genetic resources
• It has been found that in areas where regulations permit
plant collection and export, at least two years are
required to negotiate and obtain permission to collect
plant materials.

31
 Trial objectives and purpose
• The general objective is to evaluate the
safety and efficacy of African traditional
medicines (ATMs)
• The specific objectives are to conduct
preliminary safety assessments of ATMS for
specified diseases and to conduct
preliminary efficacy assessments of ATMs
for specified diseases
32
 Assessment of herbal medicines

• Assessment of quality
– Pharmaceutical assessment:
• Should cover all important aspects of
the quality assessment of herbal
medicines.
• Should make reference to a
pharmacopoeia monograph.
–if not available one must be set out
as in official pharmacopoeia.
33
• Crude plant material
– Give botanical definition.
• Genus, species and authority to ensure correct identification of
the plant.
• Definition and description of the part of the plant from which
the medicine is made (e.g., leaf, root) together with indication
as fresh, dried, or traditionally processed material is used
• The active or characteristic constituents specified, if possible
content limits defined
• Foreign matter, impurities, microbial content defined
• Voucher specimens for each lot of plant material processed,
should be authenticated by qualified botanist and stored for at
least 10 years and a lot number assigned which should appear
on the product.

34
• Plant preparations
– They include powdered plant materials,
extracts, tinctures, fatty or essential oils,
expressed juices and products resulting from
fractionation, purification or concentration.
– Manufacturing procedure should be described
in detail including substances added in the
process of manufacturing must be mentioned.
– A method of identification and assay of the
plant preparation should be added.
– If identification of an active constituent is not
possible use chromatographic finger print to
identify characteristic constituents or mixtures.

35
• Finished products
– The manufacturing procedure and formula,
including excipients should be described in
detail
– Method of identification and quantification of
plant material in finished product should be
defined otherwise use chromatographic
fingerprint to ensure consistent quality of the
preparation.
– Physical and chemical stability of the product
in the container to be marketed, storage
conditions and storage shelf life should be
established

36
• Assessment of safety
– Should cover all relevant aspects of safety assessment
– If the product has been traditionally used without
demonstrated harm
• No specific restrictive regulatory action be undertaken unless
there is new evidence of risk
– Otherwise a review of relevant literature be provided
– Monograph reference if exists, should be given
– Although long term use without evidence of risks may
indicate medicine to be harmless, it is always not
certain
• Reported side effects should be documented according to
normal pharmacovigilance practices.
• Toxicological studies if available should be part of the
assessment.
37
 Documentation of safety based on
experience
• Documentation of long term use should be
considered.
– When there are no detailed toxicological studies,
documented experiences of long-term use without
evidence of safety problems should form the basis of
the risk assessment.
• Chronic toxicological risks may have occurred but may have
not been recognized.
• The period of use, the health disorders treated, the number of
users and the countries with experiences should be
documented
• Potential for misuse, abuse, of dependence be documented

38
 Assessment of efficacy
• Review of relevant literature, original
articles and research studies
• Activity:
– The pharmacological and clinical effects of the
active ingredients and, if known, their
constituents with therapeutic activity should be
specified or described.

39
 Evaluation of efficacy and safety of herbal
medicine for the treatment of malaria.

• The general objective of these guidelines is to standardize

the methodology for documenting the ethnomedical
evidence and clinical evaluation of anti-malarial traditional
medicine in order to improve the quality and comparability
of clinical trials in this field.
• The specific objectives are to determine the efficacy of
traditional herbal medicines when given orally in the
treatment of uncomplicated malaria and determine their
safety.

40
 Case reports (ethnomedical evidence)
• Study site
– Any place where traditional herb remedies are used for
the treatment of malaria.
• Note that level of immunity of the population varies widely
from area to area, according to the level of malaria
transmission.
– Record the level of endemicity of malaria in the area as
background information.
– Record whether it is rural of urban
– Approximately what percent of population use
traditional herbal remedies as a first line treatment for
malaria

41
 Herbal medicines to be tested
• Guided by ethno-botanical information and
overview of literature
• If the traditional herbal remedy has traditionally
been used according to a credible informant with
anecdotal evidence of safety and efficacy, initial
observational studies can be carried out.
• Traditional health practitioner’s observation on
safety in pregnant women and children
– should be considered when deciding whether or not to
exclude them.
42
• The observation of patients taking these traditional herbal
remedies serve as an initial screening to determine whether
further laboratory and clinical studies would be warranted
• The following information about the medicine should be
recorded
– Name of the product
– Evidence of traditional use
• List ingredients with scientific names, part of the plant used and
quantity
• Where and when plant collected
• How the remedy is prepared, stored and administered
– In order to preserve the intellectual property rights, the above need
not be published. Copies retained by the principal investigator and
informant for reference.
• The traditional herbal remedy given to subjects must be
prepared according to the same method.
43
 Ethical considerations
• The case reports should be conducted under
the supervision of qualified staff.
– Priority at all times should be on the welfare of
subjects enrolled in the study
– Trained staff e.g. pharmacists, nurses, medical
assistants could carry out case reports.
• Subjects should be asked for their informed
consent and monitored closely.
44
 Intellectual property rights
• It is important at this stage to define who
owns the intellectual property rights to the
remedy e.g. individual health practitioner, a
group of health practitioners, a whole
community or ethnic group.
– Steps should be taken to protect their
intellectual rights, with view to drawing up a
benefit-sharing agreement should the research
proceed further.

45
 The test procedure
• Clinical assessment before commencing treatment
• Laboratory investigations
• Inclusion criteria
• Follow-up
– Administration of a traditional herb remedy according to
traditional regime for the duration of the study and ensuring that
the patient takes the right dose
– Ask about other concomitant treatment and exclude if medicines
which might alter the response are being taken (drug interactions)
• Outcome measures
– Adequate clinical response = absence of parasitaemia on day 14.
– Side effects reported
– Withdrawal from study

46
 Cohort (prospective) (Phase I)
studies
• Should be carried out on the basis of good
results from case studies or other
convincing ethnomedical evidence
• The herbal preparation should undergo
quality control assessment, and the dose be
standardized according to previous
observations

47
 Study site
• Any place where traditional herbal remedies
are used for the treatment of malaria
• The following characteristics are necessary:
– Population should live close enough to return
for follow up and sufficient numbers of malaria
patients for the study in the given time frame.
– Basic clinical and diagnostic facilities to enable
diagnosis of malaria and treatment of patients.
48
 Test system
• Same as for case reports with the following extra
considerations:
– Sample size
• Initial sample of 16
– If 0 failures, treatment is promising and requires further
investigation
– If >5 failures, there must be at least 25% failures
• Sample size of 42 is necessary if there were 1-5 failures in the
original group of 16
– <6 failures = low failure rate → move to randomized controlled
trial
– >6 failures = high failure rate → investigate alternative
preparations or dose.
• Explore about side effects in some detail.

49
 Randomized controlled trial
• Randomized clinical trial is for evaluation whether
a traditional herbal medicine for malaria is safe,
and of equivalent efficacy to the standard
recommended first-line medicines for treatment of
uncomplicated malaria
– Justification:
• Good evidence of efficacy based on ethnomedical information,
cohort study and/or convincing laboratory data
– Hypothesis:
• The traditional herbal remedy will be safe and of equivalent
efficacy to the recommended local first-line treatment for
uncomplicated malaria

50
 Drug administration.
pre-requisite for a randomized clinical trial
• A standard preparation administered orally
according to a regimen defined by ethnomedical
observation
• Sustainable supply of the medicinal plant raw
material, and conservation measures
• Laboratory data to demonstrate lack of toxicity:
cytotoxicity, teratogenicity, acute sub-acute and
sub-chronic toxicity, LD50 mouse and guinea pig
or maximum tolerated dose
• Clinical data to demonstrate safety and tolerability
in humans.
51
 Personnel and their roles
• Clinician and Traditional health practitioner:
clinical data collection and analysis.
• Experienced microscopist: counting parastaemias
• Pharmacologist/Pharmacist: pharmaceutical
formulation, pharmacology, quality control
assurance and in vitro drug sensitivity tests.
• Statistician: statistical data analysis.
• Chemist: Medicinal phytochemistry.

52
Protocol for evaluation of traditional
medicines against HIV/AIDS
• The general objective is to evaluate the efficacy and safety of
traditional medicines used for the treatment of HIV/AIDS
• Specific objectives are:
– To confirm the biological and clinical diagnosis
– To determine the clinical efficacy of the herbal product
– To document any side-effects related to the use of the product
– To determine changes in the HIV-related symptoms
– To determine changes in CD4 and CD8 counts
– To provide clinical, social and psychological support for people
living with HIV/AIDS
– To determine changes in the viral load during therapy with the
product.

53
 Criteria for patient selection
• Inclusion criteria
– Male or female age >15years
– HIV infection documented by licensed ELISA confirmed by:
– Western Blot or positive HIV blood culture
– Positive HIV serum antigen
– Second antibody test positive by a method other than ELISA
• CD4+cell count >100/mm within 2 weeks prior to entry
• Ability to provide written informed consent to
participate in the trial.
• Ability to comply with the protocol

54
 Exclusion criteria
• Previous treatment with any antiretroviral medication
• Life threatening opportunistic disease/infection
• A history of lymphoma
• Patient is a pregnant woman, or breastfeeding, child
bearing age, not on adequate birth control.
• Peripheral neuropathy
• Treatment with radiotherapy or cytotoxic
chemotherapeutic agent within 4 weeks
• Treatment with immunomodulating agents
• Active alcohol or drug abuse
• Severe kidney and/or and liver dysfunction

55
 Study site
• Two sites are required for study:
– The primary site it the traditional health
practitioner’s facility where people living with
HIV/AIDS receive treatment by the THP
• Collection of biological samples may also be done
at THP’s facility
– The secondary site is the conventional/western
health facility where laboratory investigations
are done.

56
 Traditional medicine to be used for
the study
• Should be guided by ethno botanical information
and a overview of the literature.
• If herbal remedy has been traditionally used
according to credible informant with anectode
evidence of safety and efficacy.
• The traditional health practitioner’s observations
on safety in pregnant women and children should
be taken into consideration in deciding whether or
not to exclude these groups.
57
The following information about the
medicine shall be recorded
• Name of the product
• Information on the traditional use of the product
for treating people living with HIV/AIDS
• Who owns the intellectual property rights to the
remedy
– Steps should be taken to protect their intellectual
property rights
– No information should be published

58
 Memorandum of understanding
(MOU)
• The THP and the other researchers should have developed
a memorandum of understanding before commencement of
the study.
• Such an MOU would specify the responsibilities of
stakeholders including secrecy of data by members of the
research team.
• Dose schedule and administration:
– The preparation will be used as presented by the THP who
formulates, prescribes and dispenses the product to the
participants.
– The commencement of the study is subject to approval of
appropriate national authority e.g. Food and drug administration.

59
 Intellectual property rights
• A legal agreement between the THP and the
scientists should among others, clearly specify the
responsibilities for patency or securing trade
secrets on the products, ownership of the patents,
trade secrets, royalties initial lump sum payment
for indigenous knowledge, plant collection etc.
• There should be regal protection before the MOU
is effected.

60