DRUG DEVELOPMENT PROCESS

INTRODUCTION :

Drug development involves a series of scientific, preclinical, and clinical studies designed
to evaluate a compound’s safety, efficacy, quality, dosage, and therapeutic value,
followed by regulatory approval before it can be marketed for patient use.

During the development, the new drug or the compound should adhere to high standards
in the conduct, analysis and interpretation of preclinical and clinical studies for its smooth
passage through the regulatory approval phase and eventually to marketing. The drug
discovery and drug development process is designed to ensure that only those
pharmaceutical products that are both safe and effective are brought to market.

The following sequence of research activities begins the process that results in
discovery of new medicines:
1. Target Identification
2. Target Validation
3. Lead identification
4. Lead Optimization

Drug development phases :- There are three major phases in drug development:

1) Pre-clinical research and development

2) Clinical research and development
3) After the compound is on the market, a possible “post marketing”
phase
The pre-clinical phase represents bench (in vitro) and then animal testing, including
kinetics, toxicity and carcinogenicity. In the U.S., an investigational new drug
application (IND) is submitted to the Food and Drug Administration seeking
permission to begin the heavily regulated process of clinical testing in human
subjects. The clinical research (IND) phase representing the time from beginning of
human trials to the new drug application (NDA) submission that seeks permission to
market the drug is by far the longest portion of the drug development cycle and can
last from 2 to 10 years. Phase I trials, sometimes called, “first in human” trials, are
generally conducted on relatively small groups (typically 10 to 30) of health
VDlunteers (except for oncology drugs or other potentially toxic compounds) in
specialized units resembling small hospitals with 20 to 50 monitored beds. The
“inpatient” portion of Phase I trials usually lasts from a day or two to a week (though
follow up can lastup to about a month), and are designed to assess the safety of a
compound and study its pharmacokinetics and pharmacodynamics.
Assuming a compound is shown to be safe for healthy subjects and survives Phase I,
then development proceeds to a series of Phase II trials. These trials typically enroll
anywhere from about 20 or 30 patients up to a few hundred at most. These patients
usually have relatively “pure” form of the disease for which the drug is intended. In
other words, they suffer from as little other intercurrent disease as possible, and the
list of concomitant medications they can be taking is usually restricted. For example,
patients with newly diagnosed, but untreated, diabetes, with no evidence of end
organ damage, would be used to test a new
antidiabetic agent. Phase II trials tend to last only a few weeks to, at most, a few
months. Initial Phase II trials (sometimes called, Ila) are pilot trials to determine dose
range.
They tend to be conducted at specialized centers, like university medical centers, by
specialized investigators, such as medical school faculty For example, patients with
newly diagnosed, but untreated, diabetes, with no evidence of end organ damage,
would be used to test a new antidiabetic agent. Phase II trials tend to last only a few
weeks to, at most, a few months. Initial Phase II trials (sometimes called, Ila) are pilot
trials to determine dose range. They tend to be conducted at specialized centers, like
university medical centers, by specialized investigators, such as medical school
faculty. Phase III trials are conducted on larger group of population. Phase IV trails
which are called as Post- marketing phase trails are conducted añer the drug has been
introduced into the market.

APPROACHES OF DRUG DISCOVERY :

Pharmacoloqical Approach:

The first step in the process of drug discovery is to identify a molecular target that is
associated with a particular disease. The next step is to establish that modulation or
intervention of the function of the chosen target is associated with a reversal of the
disease process. The research in integrative cell signaling and neuroscience programs
in the Department of Pharmacology offers a wealth of novel molecular targets to
consider for therapeutic intervention. Once a particular target is selected, a
number of
approaches may be taken to modulate or interfere with the biological function. These
approaches might include the use of small molecules or protein biologicals such as
monoclonal antibodies that alter biological activity. These efforts are aided by the Yale
West Campus screening facilities: High Throughput Cell Biology (HTCB) that offers
screening with RNAi and the Small Molecule Discovery Center that provides access to
libraries of unique small molecules. We work closely with the Structural Biology program
in Pharmacology to obtain the detailed three- dimensional structure of the target along with
novel ligands as lead compounds that affect function. Structural biology also assists in lead
optimization to obtain candidate compounds that are suitable for further investigation.
Possible candidate molecules are further examined in preclinical studies and the most pro
missing compounds would ultimately be examined in clinical trials in humans. Our
department offers a number of courses to provide an understanding of how the drug
discovery process works from the identification of molecular targets and therapeutic
candidates to the design of clinical trials in humans
As compounds progress through the Athersys tier system, we have obtained information
related to compound specificity, selectivity, half-life and disposition in animals,
toxicological properties, and efficacy in disease models. Athersys has applied a rigorous
pharmacological testing, including testing for effects on liver enzymes, specific cardiac
channels, and a large number of unrelated targets where interactions might be problematic.
In addition, the company has evaluated the effects of compounds on animal behavioral
endpoints, organ pathology and cardiovascular
 Drug Discovery and Development
function to further increase confidence that a compound will be safe and efficacious.

Toxicoloqical Approach :

Although basic toxicology parameters will have been assessed in lead optimization
series of in vitro and in vivo studies will be performed in the development phase,
usually to registration standards and protocols, in order to fully determine the toxicity
profile and therapeutic index, maximum doses for safe administration etc. These will
usually be performed in at least two species and should determine both specific
(mechanism related) and non-specific (chemical) side effects which could limit the
safe dose which can be used in man. If the mechanism is human specific and doesn't
occur in man these studies will be limited to non-specific toxicology with ex-vivo
human studies etc being implemented to assess the mechanism related effects. The
duration and number of dose levels used in these studies will be determined by the
drug exposure, length and protocols of the proposed initial (Phase I/II) clinical studies
in man.
Studies will also be performed to assess the broader safety effects such as behavioral
and physiological effects in vivo.
For the most part, the current approaches to toxicity testing and safety assessment rely
on a complex array of traditional studies that evaluate observable outcomes in whole
animals, such as clinical signs or pathological changes that are indicative of a disease
state. As noted by MacDonald and Robertson, “The approaches that have been
taken to assess human risk of
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 Drug Discovery and Development
adverse effects from chemical exposure have changed very little over the last several
decades.” For example, "The global regulatory requirements for registration of new
human pharmaceutical chemicals—the data requirements have changed very little since
their establishment three and sometimes four decades ago despite the dramatic
advances in the sciences that are used for this activity.”

1. Most of the toxicology tools used for regulatory assessment rely on high-dose
animal studies and default extrapolation procedures, and have remained
relatively unchanged for decades, despite the scientific revolutions of the past
half- century.
2. Indeed, when the FDA released its strategic plan in August 2011, the need to
modernize toxicology to enhance product safety was the first of eight science
priority areas identified

Druq characterization:

It is recognized that drug candidates with drug-like properties are more likely to make
into market. Thus drug like property assessment has been integrated into the current
drug discovery and development process.
Research has shown that these physico-chemical properties are closely related to
drug's absorption, distribution, metabolism and excretion (ADME). They, together
play important roles in the absorption and bioavailability of drug candidates. Early
assessments of these properties in drug discovery can help

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 Drug Discovery and Development
project teams find potential PK issues, optimize chemical series and avoid promoting
“problematic" compounds to development
Drug Characterization involves finding of biological and
physiochemical properties of potential drug molecules.

• Biological Characterization
Interaction of drug molecule with Biological systems Ex- Receptor
binding, Enzyme inhibition
• Physiochemical Characterization
Physical and chemical characteristics of potential drug Ex- Soluability,
PKa
X-ray, NMR,IR,UV-VIS,MS,PET

Biological Characterization:

It is needed to assess whether the molecule will likely owork in humans.

Goals :

1. To find Efficacy
2. Asses Toxic Effects
Further Divided into

A. Biochemical Assay
B. Cellular Assay
C. System/Whole Animal assay

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 Drug Discovery and Development

A. Biochemical Assay
Mechanism of action at molecular level
Receptor binding assay's (Affinity and selectivity) Metabolic
Processing

B. Cellular Assay
Cellular Cultures: Cell penetration Receptor activity,
Transport
Isolated Tissues Effects on specific tissues,PossibIe toxicity
» Liver tissues : Drug metabolism drug interaction

C. System/Whole animal Assay

Rat,Mouse : Toxicity, LD50, Organ-Specific efficacy
Dog,Rabbit,Guinea pig, Monkey —
Pharmacokinetics/Dynamics, BA,Toxicity.
Lab Animal Models for disease: Efficacy and toxicity

& Physiochemical Characterization

Physical and chemical properties of a drug are critical for knowing how it can be
formulated/stored/administered
Solubility ñ*
PKa
Partition coefficient: Octanol: water
\ Stability

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 Drug Discovery and Development
» Chemical Structure: Mass Spectroscopy, NMR,X-Ray
Crystallography
Impurities : TLC,HPLC,GC, Mass Spectroscopy
\ Polymorphism

Dosaqe form:

Developability Assessment Supporting Drug Candidate

Selection

• Profiling of key Physicochemical Properties

• Biopharmaceutical Assessment
Pre-formulation

• Solubility, Stability, Dissolution Rate and Solid State

Properties
• Salt and Form Screening and Selection
• Excepient Compatibility
Dosage Form Design
• Conventional
•Non-Conventional
• Formulation Development, Evaluation and Scale-up

Dosage form Design: Evolution of Dosage Form Needs

• Preclinical (Toxicology) Dosage Forms
• Biopharmaceutics
• Emphasis is on Exposure, High Drug Concentration
• Suspensions (NaCMC), Drug/Feed Mixtures
• Specialized Forms (Yogui1)

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 Phase I Clinical Dosage Forms
• Biopharmaceutics - Simple, Flexible Dosing
• Single/Multiple Dose Tolerability Studies, Healthy Volunteers
• Efficacy Marker -Proof of Concept, Attrition Curve
• Solutions, Powder-in-a-Bottle, Hard Gelatin Capsules

Phase II Clinical Dosage Forms

• Biopharmaceutics, Physico-Chemical, Processing -Larger, Longer Studies
• Patients, Dose Finding and Proof of Concept
• Blinded Capsules or Tablets

Biopharmaceutical
• Mechanism of Action,Target Organ,Dose (Potency)PermeabiIity
(Passive, Active, Efflux)
• Pharmacokinetics
Absor

SCHEDULE Y

PHARMACOLOGICAL AND TOXICOLOGICAL STUDIES

Introduction

Schedule Y contains requirements and guidelines for permission to import

and/or manufacture of new drugs for sale or to undertake clinical trials.

There are 12 appendices in schedule Y

Appendix 3: Animal Toxicology

Appendix 4: Animal Pharmacology

Appendix III: Animal toxicology (Non-clinical Toxicity studies)

General Principles

 should comply with the norms of Good Laboratory Practice (GLP)

 should be performed by suitably trained and qualified staff

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 properly calibrated and standardized equipment of adequate size and

capacity

 Standard operating procedures (SOPs) should be followed for all

managerial and laboratory tasks

 Test substances and systems should be properly characterized and

standardized.

 All documents belonging to each study (like approved protocol, raw

data, draft report, final report, and histology slides and parafÏn tissue

blocks) should be preserved for a minimum of 5 years after marketing

of the drug.

 Toxicokinetic studies should be conducted to assess the systemic

exposure achieved in animals and its relationship to dose level and the

time course of the toxicity study.

1.1 SYSTEMIC TOXICITY STUDIES

1.2 Male Fertility Study
Species: 1 rodent (preferably rat)
Route: as intended for clinical use
Dose: 3 graded doses (high, intermediate, low)
Observation: 28-70 days before pairing
Group: 3 dose groups + 1 control group; 6 adult males each
Animals are paired with female animals of proven fertility in a ratio of 1:2 for
mating.
Drug treatment of the male animals should continue during pairing.
Pairing is continued till the detection of vaginal plug or 10 days, whichever is
earlier.

Aysha Shaharbanu, Asst professor 11

Females getÝng thus pregnant should be examined for their fertility index
after day 13 of gestation.
All the male animals should be sacrificed at the end of the study.
Weights of each testis and epididymis should be separately recorded.
Sperms from one epididymis should be examined for their motility and
morphology. The other epididymis and both testes should be examined for
their histology.
1.3 Female Reproduction and Developmental Toxicity Studies
These studies need to be carried out for all drugs proposed to be studied or
used in women of child bearing age.
1.3.1 Female Fertility Study (Segment I):
Species: 1 rodent (rat preferred)
Route: As intended for clinical use
Dose: 3 Graded doses (Males: 28 days & Females: 14 days before mating,
continued during mating and gestation)
Group: 3 dose groups + control group; 15 males and 15 females per group
Dams should be allowed to litter and their medication should be continued till
the weaning of pups.
Observation parameters
For dams:
 Body weight,
 food intake,
 clinical signs of intoxication,
 mating behavior,
 progress of gestation/ parturition periods,
 length of gestation,
 parturition,
 post-partum health,
 gross pathology (and histopathology of affected organs) of dams.
For pups (both treated and control groups):
 General signs of intoxication,
 sex-wise distribution in different treatment groups,

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 body weight,
 growth parameters,
 survival,
 gross examination,
 autopsy.
Histopathology of affected organs should be done.
1.3.2 Teratogenicity Study (Segment II):
Species: 1 Rodent (preferably rat) and 1 Non-rodent (rabbit)
Route: As intended for clinical use
Dose: 3 Graded doses (continued throughout organogenesis)
Group: 3 dose groups + control group; 20 pregnant rats (or mice) and 12
rabbits
All fetuses should to be subjected to gross examination, one half of the fetuses
should be examined for skeletal abnormalities and the other half for visceral
abnormalities.
Observation parameters
For dams:
 signs of intoxication,
 effect on body weight,
 effect on food intake,
 examination of uterus,
 ovaries and uterine contents,
 number of corpora lutea,
 implantation sites,
 resorptions (if any)

4. increasing group size with increase in duration of treatment.

1.5 Allergenicity/ Hypersensitivity:
Standard tests include Guinea Pig Maximization Test (GPMT) and Local Lymph
Node Assay (LLNA) in mouse.
Any one of the two may be done.
(i) Guinea Pig Maximization Test:
STEP 1: Determination of maximum nonirritant and minimum irritant dose
Aysha Shaharbanu, Asst professor 13
Intradermal induction dose is determined using 4 doses in 4 males and 4
female animals.
Topical minimum irritant dose is determined using 2 males and 2 females.
STEP 2: Main Test
One test and one control group, with a minimum of 6 males and 6 female
animals in each group.
Intradermal induction (day 1) coupled with topical challenge (day 21) should be
done.
If there is no response, re-challenge should be done 7-30 days after the
primary challenge.
Erythema and edema should be used as evaluation criteria.
(ii) Local Lymph Node Assay:
Mice used in this test should be of the same sex, either only males or only
females.
Drug treatment is to be given on ear skin. Three graded doses, the highest
being maximum nonirritant dose plus vehicle control should be used.
A minimum of 6 mice per group should be used.
Test material should be applied on ear skin on three consecutive days and on
day 5, the draining auricular lymph nodes should be dissected out 5 hours
after i.v. H-thymidine or bromo-deoxy-uridine (BrdU).
Increase in H-thymidine or BrdU incorporation should be used as the criterion
for evaluation of results.

[Link] Urinary System

Urine volume, specific gravity, osmolality, pH, proteins, cytology and blood
urea nitrogen, creatinine and plasma proteins estimation.
[Link] Autonomic Nervous System
Binding to receptors relevant for the ANS, and functional response to
agonist or antagonist responses in vivo or in vitro
[Link] Gastrointestinal System
Studies on gastric secretion, gastric pH measurement, gastric mucosal
examination, bile secretion, gastric emptying time in vivo and ileocecal
contraction in vitro.
Aysha Shaharbanu, Asst professor 14
 [Link] Other Organ Systems
Effects of the investigational drug on organ systems not investigated
elsewhere should be assessed when there is a cause for concern. For
example dependency potential, skeletal muscle, immune and endocrine
functions
1.4 Conditions Under Which Safety Pharmacology Studies Are Not Necessary
Safety pharmacology studies are usually not required for:
 locally applied agents e.g. dermal or ocular,
 cases where the pharmacology of the investigational drug is well known,
and/or when systemic absorption from the site of application is low.
 new derivative having similar pharmacokinetics and pharmacodynamics.
1.5 Timing Of Safety Pharmacology Studies In Relation To Clinical Development
1.5.1 Prior to first administration in humans
The effects of an investigational drug on the vital functions listed in the
essential safety pharmacology should be studied prior to first
administration in humans.
1.5.2 During clinical development
Additional investigations may be warranted to clarify observed or suspected
adverse effects in animals and humans during clinical development.
1.5.3 Before applying for marketing approval
Follow-up and supplemental safety pharmacology studies should be
assessed prior to approval. Available information from toxicology studies
addressing safety pharmacology endpoints or information from clinical
studies can replace such studies.

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