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Objective: The present study was carried out to assess the safety and efficacy of two Metformin
Hydrochloride (SR) 500 mg tablet formulations. The test formulation was Metformin
Hydrochloride (SR) 500 mg tablet of Vapi Care Pvt. Ltd., India, and the reference standard was
Dibeta SR tablet [containing Metformin Hydrochloride (SR) 500 mg] of Torrent Pharmaceuticals
Ltd., India.
Methodology: The present study was an open label, balanced, randomized, two treatment, two
period, two sequence, single dose, cross over bioequivalence study under fasting condition;
conducted at Auriga Research Ltd., New Delhi, India. Metformin Hydrochloride (SR) 500 mg
tablet of Vapi Care Pvt. Ltd., India, was the Test formulation and Dibeta SR tablet [containing
Metformin Hydrochloride (SR) 500 mg] of Torrent Pharmaceuticals Ltd., India, was the
Reference standard. Hippocrates Independent Ethics Committee (HIEC), New Delhi, India was
the ethical committee for this study. There was no dropout/withdrawal of subjects from this
study. Subjects were randomly assigned to receive a single oral dose of the test and the reference
formulation under fasting condition, with a washout period of seven days. Blood samples were
drawn as per the approved study protocol. Drug concentration in the plasma samples were
measured by using validated LC/MS/MS method. Win Nonlin Version 5.2 software was used for
statistical calculations.
Results and Discussion: The 90% confidence interval for ln-transformed pharmacokinetic
parameters Cmax, AUC0-t and AUC0- of the test formulation was 89.13%, 87.46% and 88.29%,
respectively. The 90% confidence interval for ln-transformed pharmacokinetic parameters Cmax,
AUC0-t and AUC0- of the reference standard was 112.44%, 123.85% and 123.87%, respectively.
The mean test/reference ratio for ln-transformed Cmax, AUC0-t and AUC0- were 100.110%,
104.080% and 104.580%, respectively.
Conclusion: Ln-transformed pharmacokinetic parameters Cmax, AUC0-t and AUC0- of the test
formulation and reference standard, are within the acceptable range of 80125% (to prove
bioequivalence between two drugs). So, the present study concludes that the test formulation
Metformin Hydrochloride (SR) 500 mg of Vapi Care Pvt. Ltd., India is bioequivalent to the
reference standard Dibeta SR tablet [containing Metformin Hydrochloride (SR) 500 mg] of
Torrent Pharmaceuticals Ltd., India.
LIST OF ABBREVIATIONS
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AE Adverse Event
BA Bioavailability
BE Bioequivalence
BP Blood Pressure
CI Confidence Interval
cm/s Centimeter/s
CV Coefficient of variation
CYP Cytochrome P
dl Decilitre
ECG Electrocardiogram
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gm Gram
h / hr/ Hr Hour
HCT Haematocrit
Ht Height
IgE Immunoglobulin E
IP Investigational Product
IU International Units
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IV Interavenous
Kg Kilogram
L Litre
m Metre
g Microgram
l/ul Microlitre
mEq Milliequivalent
mg Milligram
ml Millilitre
min Minute/Minimum
NA Not Applicable
ng Nanogram
No. Number
P Probability
pg Picogram
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PK Pharmacokinetic
QC Quality Control
R Reference
SD Standard Deviation
SE Standard Error
T Test
THC Tetrahydrocannabinoids
U Unit
UK United Kingdom
Wt Weight
C Degree Celsius
% Percentage
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List of Tables
24
1 2x2 crossover design
13 Types of Insulin 46
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17 Randomization Schedule 72
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18 Phlebotomy Schedule 73
89-90
Urine Analysis
23
24 91
Blood Sample Collection
30 100
Deviations in Vital Signs Measurements
31 101
Blood glucose monitoring Period I
32 102
Blood glucose monitoring Period II
33 103
Incidence of Adverse Events
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List of Figures
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Contents
1 Introduction 1-4
2 Objectives 5
4 Methodology 62-85
6 Conclusion 107
7 References 108-110
8 Annexures 111-123
1. Introduction
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Bioavailability era was started as early as 1945, with the first publication of the concept
of biological availability. The development of analytical techniques, during the 1960s,
made possible the development of methods sensitive enough to allow quantification of
drugs or metabolites, initially in the urine and afterwards in the plasma, making
possible the evaluation and the comparison of bioavailability of different formulations,
as well as demonstration that significant differences among them could occur.
Bioavailability is defined as the rate and extent to which the active ingredient or active
moiety is absorbed from a drug product and becomes available at the site of action. For
drug products that are not intended to be absorbed into the bloodstream, bioavailability
may be assessed by measurements intended to reflect the rate and extent to which the
active ingredient or active moiety becomes available at the site of action. (FDA,2003).
The FDA usually considers that the plasma concentration of a drug is a surrogate for
the concentration at the site of action for a systemically acting drug. Proving
equivalence requires integration of several studies, such as PK, PD, controlled-clinical,
in vitro studies, and any other specific model or study that may prove useful in proving
equivalence.
The Drug Price Competition and Patent Term Restoration Act of 1984 (the Hatch
Waxman Amendments), established the current Abbreviated New Drug Application
approval process. The showing that must be made for an Abbreviated New Drug
Application to be approved is quite different from what is required in an New Drug
Application. An New Drug Application applicant must prove that the drug product is
safe and effective. An Abbreviated New Drug Application does not have to prove the
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safety and effectiveness of the drug product, because an Abbreviated New Drug
Application relies on the finding the FDA has made that the reference listed drug is safe
and effective. Instead, an Abbreviated New Drug Application applicant must
demonstrate, among other things,that its drug product is bioequivalent to the reference
listed drug. The scientific premise underlying the HatchWaxman amendments is that
in most circumstances, bioequivalent drug products may be substituted for each other.
A generic drug is bioequivalent to the listed drug if the rate and extent of the
absorption of the drug do not show a significant difference from the rate and extent of
absorption of the listed drug when administered at the same molar dose of the
therapeutic ingredient under similar experimental conditions in either a single dose or
multiple doses.
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Globally diabetes affects 246 million people and is expected to affect 366 million by
2030. At least 7 million new cases are reported annually. India accounts for largest
diabetic population with 41 million patients, mounting to 6 percent of the adult
population. [3]
Various classes of oral hypoglycaemics are available for the treatment of Type 2
diabetes mellitus in addition to insulin formulations. These are Sulfonylureas,
Biguanides, Alpha-Glucosidase Inhibitor, Thiazolidinediones (TZD), Meglitinides.
Amylin Analogues, Incretin Mimetics and Dipeptidyl Peptidase-4 (DPP-4) Inhibitors
are the new treatment regimens available for the Type 2 diabetes mellitus. As per the
Global Guideline for Type 2 Diabetes (developed by International Diabetes
Federation, 2005), Metformin is considered as a first line drug.
Metformin is the only biguanide available in the market. Metformin has been used
clinically for 45 years, and has been approved since 1995. Metformin has become the
first-line therapy for glycemic control when oral agents are indicated in overweight
patients. The action of metformin does not involve stimulation of pancreatic insulin
secretion and therefore it is still a beneficial agent when -cell function has declined.
Another advantage of metformin over insulin secretagogues, and sulfonylureas in
particular, is that it does not usually cause hypoglycemia and weight gain. Metformin
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has a short duration of action, with a half-life of between 1.3 and 4.5 hours, and does
not bind to plasma proteins. It is not metabolised and is totally renally eliminated.
After the patent expires, several companies may manufacture and market different
formulations of a same active substance, with similar qualities and performances, so as
the interchangeability among different formulations, when given in equivalent doses,
presents the same safety and efficacy. Generic substitution has thus provided a means
of supplying the market with inexpensive, efficacious, and safe drug products after
patent expiration, without repeating the entire drug development process. [5]
Thats why Sponsor tried to bring a new generic version of Metformin Hydrochloride
(SR) 500 mg tablet into the market. As per the regulation, bioequivalence study should
be done before marketing. So, a bioequivalence study was conducted at Auriga
Research Ltd, comparing the Test formulation Metformin Hydrochloride (SR) 500 mg
tablet of Sponsor with the Reference standard Dibeta SR tablet [containing Metformin
Hydrochloride (SR) 500 mg] of Torrent Pharmaceuticals Ltd., India.
This was a DCGI submission study, so the CDSCO BA/BE Guidelines, 2005 were
followed in conjunction with Schedule Y to the Drugs and Cosmetic Rules, Indian GCP
Guidelines issued by CDSCO, GLP and the Ethical Guidelines for Biomedical
research on human subjects issued by Indian Council of Medical Research (ICMR
Guidelines). ICH-GCP Guidelines and Declaration of Helsinki were also referred as
and when required.
2. Objectives
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3. Review of Literature
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This figure shows in an idealized way the stages of a typical project, aimed at
producing a marketable drug that meets a particular medical need (e.g. to retard the
progression of Parkinsons disease or cardiac failure, or to prevent migraine attacks).
Broadly, the process can be divided into three main components, namely:
Drug Discovery, during which candidate molecules are chosen on the basis of
their pharmacological properties.
Preclinical Development, during which a wide range of non-human studies
(e.g. toxicity testing, pharmacokinetic analysis and formulation) are performed.
Clinical Development, during which the selected compound is tested for
efficacy, side effects and potential dangers in volunteers and patients.
These phases do not necessarily follow in strict succession as indicated in the above
figure, but generally overlap.
Clinical development proceeds through four distinct phases (Friedman et al., 1996)
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3.2.1 Phase I
Phase I trials are performed on a small group (normally 20-80) of normal healthy
volunteers, and their aim is to check for safety (does the drug produce any potentially
dangerous effects, for example on cardiovascular, respiratory, hepatic or renal
function?), tolerability (does the drug produce any unpleasant symptoms, for example
headache, nausea, drowsiness?) and pharmacokinetic properties (is the drug well
absorbed? What is the time course of the plasma concentration? Is there any evidence
of cumulation or non-linear kinetics?). Phase I studies may also test for
pharmacodynamic effects in volunteers (e.g. does a novel analgesic compound block
experimentally induced pain in humans? How does the effect vary with dose?).
3.2.2 Phase II
Phase II studies are performed on groups of patients (normally 100-300) and are
designed to test for efficacy in the clinical situation, and if this is confirmed, to
establish the dose to be used in the definitive Phase III study. Often, such studies will
cover several distinct clinical disorders (e.g. depression, anxiety states and phobias) to
identify the possible therapeutic indications for the new compound and the dose
required. When new drug targets are being studied, it is not until these Phase II trials
are completed that the team finds out whether or not its initial hypothesis was correct,
and lack of the expected efficacy is a common reason for failure.
Phase III studies are the definitive double-blind randomized trials, commonly
performed as multicentre trials on 1000-3000 patients, aimed at comparing the new
drug with commonly used alternatives. These are extremely costly, difficult to
organize, and often take years to complete, particularly if the treatment is designed to
retard the progression of a chronic disease. It is not uncommon for a drug that seemed
highly effective in the limited patient groups tested in Phase II to look much less
impressive under the more rigorous conditions of Phase III trials.
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Clinical Practice, covering every detail of the patient group, data collection methods,
recording of information, statistical analysis and documentation.
At the end of phase III, the drug will be submitted to the relevant regulatory authority
for licensing. The dossier for this is a massive and detailed compilation of preclinical
and clinical data. Evaluation by the regulatory authority normally takes a year or more,
and further delays often arise when aspects of the submission have to be clarified or
more data are required. Eventually, about two-thirds of submissions gain marketing
approval.
3.2.4 Phase IV
Phase IV studies comprise the obligatory post marketing surveillance designed to detect
any rare or long-term adverse effects resulting from the use of the drug in a clinical
setting in many thousands of patients. Such events may necessitate limiting the use of
the drug to particular patient groups, or even withdrawal of the drug. [6]
The process of movement of drug from its site of administration to the systemic
circulation is called as absorption. Bioavailability is defined as the rate and extent
(amount) of drug absorption. The movement of drug between one compartment and the
other (generally blood and the extravascular tissues) is referred to as drug distribution.
Elimination is defined as the process that tends to remove the drug from the body and
terminate its action. Elimination occurs by two processes-biotransformation
(metabolism), which usually inactivates the drug, and excretion which is responsible for
the exit drug/metabolites from the body.
A direct relationship exists between the concentration of drug at the biophase (site of
action) and the concentration of drug in plasma. A typical plasma drug concentration-
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time curve obtained after a single oral dose of a drug and showing various
pharmacokinetic and pharmacodynamic parameters is depicted in Fig.2
The three important pharmacokinetic parameters that describe the plasma level-time
curve and useful in assessing the bioavailability of a drug from its formulation are:
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The point of maximum concentration of drug in plasma is called as the peak and the
concentration of drug at peak is known as peak plasma concentration. It is also called
as peak height concentration and maximum drug concentration. Cmax is expressed in
mcg/mL.
The peak represents the point of time when absorption rate equals elimination rate of
drug. The portion of curve to the left of peak represents absorption phase i.e. when the
rate of absorption is greater than the rate of elimination. The section of curve to the
right of peak generally represents elimination phase i.e. when the rate of elimination
exceeds rate of absorption.
The time for drug to reach peak concentration in plasma (after extravascular
administration) is called as the time of peak concentration. It is expressed in hour (hr)
and is useful in estimating the rate of absorption. Onset time and onset of action are
dependent upon tmax.
It represents the total integrated area under the plasma level-time profile and expresses
the total amount of drug that comes into the systemic circulation after its
administration. AUC is expressed in mcg hr/mL. It is the most important parameter in
evaluating the bioavailability of a drug from its dosage form as it represents the extent
of absorption.
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It is the time required for the drug to start producing pharmacologic response. It
corresponds to the time for the plasma concentration to reach MEC after administration
of drug.
v) Duration of Action
The time period for which the plasma concentration of drug remains above the MEC
level is called as duration of drug action.
The drug concentration between MEC and MSC represents the therapeutic range. [7]
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The first and most important measure assessed is the area under the curve of the drug
plasma concentration versus time, often used to measure absorption length or the total
amount of drug absorbed by the body after a single-dose administration of a
medication. The determination of bioequivalence between two medications results from
the comparison of AUCs obtained from the experiment. By its mathematical
representation
Among several methods for the determination of AUC from zero time to last collection
time (tk) the most used one is that of trapezoids (Chow & Liu, 2006). This method
comprises the sum of trapezium areas as determined by collection times and respective
concentrations.
Let us assume that C0, C1, C2, Ck, are the concentrations obtained in an experiment for
collection times 0, t1, t2, tk, respectively. AUC from zero to tk, denoted by AUCtk, is
obtained as follows:
The area under the curve of concentration versus time (AUC) may also be extrapolated
and calculated from zero time up to the time related to the complete elimination of the
drug. This measure is referred to in literature as the area under the curve from zero time
to infinite. The additional portion is expressed by a relationship between the last
concentration measured Ck and the drug elimination speed constant Ke. The
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Apparent Volume of Distribution = Amount of the drug in the body / Plasma drug
concentration
i.e. Vd = X / C
Drugs that are highly lipid soluble, such as digoxin, have a very high volume of
distribution (500 litres). Drugs, which are lipid insoluble, such as neuromuscular
blockers, remain in the blood, and have a low Vd.
Clearance (Cl) is defined as the theoretical volume of body fluid containing drug (i.e.
that fraction of apparent volume of distribution) from which the drug is completely
removed in a given period of time. It is expressed in ml/min or liters/hour.
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i.e. Cl = dX/dt/C
Elimination half-life (t1/2) is defined as the time taken for the amount of drug in the
body as well as plasma concentration to decline by one-half or 50% its initial value. It
is expressed in hours or minutes. It is also called as biological half-life. After four half-
lives, elimination is 93.75% complete.
t1/2= 0.693/ KE
If the Apparent Volume of Distribution (Vd) is increased, then elimination rate constant
(KE) will decrease, the Elimination half-life (t1/2) will increase, but the clearance (Cl)
will not change. [9]
First order process is one whose rate is directly proportional to the concentration of
drug undergoing reaction i.e. greater the concentration, faster the reaction. It is because
of such a proportionality between rate of reaction and the concentration of drug that a
first-order process is said to follow linear kinetics.
A constant fraction of the drug in the body is eliminated per unit time. The rate of
elimination is proportional to the amount of drug in the body. The majority of drugs are
eliminated by this way. Most of the pharmacokinetic processes like absorption,
distribution follow first order kinetics. [10]
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During its existence, a new drug, is considered to have 3 life periods. The first of them
is before its approval for public use, when pre-clinical and clinical studies are carried
out, to evaluate efficacy and safety. The second comprises the life span of its patent, in
which a marketing exclusivity is still valid. The last period begins just after the patent
expires and other companies may market the product, as a similar or as a generic drug.
Thus, several companies may manufacture and market different formulations of a same
active substance, with similar qualities and performances, so as the interchangeability
among different formulations, when given in equivalent doses, presents the same safety
and efficacy.
Bioavailability era was started as early as 1945, with the first publication of the concept
of biological availability. The development of analytical techniques, during the 1960s,
made possible the development of methods sensitive enough to allow quantification of
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drugs or metabolites, initially in the urine and afterwards in the plasma, making
possible the evaluation and the comparison of bioavailability of different formulations,
as well as demonstration that significant differences among them could occur.
Bioavailability is defined as the rate and extent to which the active ingredient or active
moiety is absorbed from a drug product and becomes available at the site of action. For
drug products that are not intended to be absorbed into the bloodstream, bioavailability
may be assessed by measurements intended to reflect the rate and extent to which the
active ingredient or active moiety becomes available at the site of action. (FDA,
2003).
The FDA usually considers that the plasma concentration of a drug is a surrogate for
the concentration at the site of action for a systemically acting drug. Proving
equivalence requires integration of several studies, such as PK, PD, controlled-clinical,
in vitro studies, and any other specific model or study that may prove useful in proving
equivalence.
The concept of Bioequivalence and the required proof by the regulatory agencies has
evolved over the past several decades:
In the United States, the 1902 federal law for biologics, particularly vaccines,
required evaluation for safety, purity, and potency.
The 1906 Food and Drugs Act added drugs other than biologics.
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The 1938 FDC Act created FDA and evaluation of new drugs based on data in a
filed NDA.
The 1962 law added effectiveness requirements for the approval of NDA. 1902
Federal law required that biologics (vaccines) be evaluated for safety, purity,
and potency.
The 1906 Food and Drugs Act added drugs other than biologics.
The 1938 FDC Act created FDA and required safety evaluation on new drugs
before marketing based on data in an NDA.
The 1962 law added effectiveness requirement for approval of an NDA; in the
1960s, the FDA permitted marketing of similars, while corresponding pioneer
products underwent
Drug Efficacy Study Implementation (DESI) reviews. Similars came into the
market between 1938 and 1962.
In 1970 the FDA terminates marketing of similars unless DESI pioneer
showed safety and efficacy.
Similar manufacturer submits ANDA with formulation and manufacture
information;
(The Supreme Court in the United States vs. Generix Drug Corporation
supported FDA requirement for ANDA).
The 1984 generic law in the United States (WaxmanHatch) created a generic
approval system for all new drugs, including those approved after 1962. The
FDA finalized the bioequivalence (BA/BE) regulations, wherein the pioneer
shows BA in NDA; similars to DESI-effective pioneers show BE leading to
first United States first generics. Several revisions to the bioequivalence
(BA/BE) regulations, were made including the most recent one in April 2006.
The Drug Price Competition and Patent Term Restoration Act of 1984 (the
HatchWaxman Amendments), established the current Abbreviated New Drug
Application approval process. The showing that must be made for an
Abbreviated New Drug Application to be approved is quite different from what
is required in an New Drug Application. An New Drug Application applicant
must prove that the drug product is safe and effective. An Abbreviated New
Drug Application does not have to prove the safety and effectiveness of the
drug product, because an Abbreviated New Drug Application relies on the
finding the FDA has made that the reference listed drug is safe and effective.
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A generic drug is bioequivalent to the listed drug if the rate and extent of the
absorption of the drug do not show a significant difference from the rate and extent of
absorption of the listed drug when administered at the same molar dose of the
therapeutic ingredient under similar experimental conditions in either a single dose or
multiple doses.
3.5.1Clinical Step
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The performance of a literature research is the first step for the search of the
bioanalytical method. Once the method exists, it should be tested for its reproducibility.
In the event there is no bioanalytical method for a given drug, the analytical center
should develop a method that responds satisfactorily to the study aimed.
The previous performance of the steps required for the development of the analytical
method for bioequivalence studies assures to the analytical center and its contractor that
the services hired will be carried out at the deadline foreseen and with the required
reliability of the results, which will be evaluated for registration purposes of the study
drug. In this context, we may state that contracted party and contractor will not waste
their time and additional financial resources in the event the studies performed are
rejected in the end due to the inappropriateness of the method used and undetermined
storage conditions.
Concentration levels of a drug in the body depend partially on the administration route,
which may be classified as intravascular or extravascular. Intravascular administration
is carried out directly into blood stream by intravenous or intra-arterial route.
Extravascular includes oral, intramuscular, subcutaneous, transdermal and other
administration routes. When a drug is administered to the body, it generally passes
through the absorption, distribution, metabolism, and finally, elimination stages. Thus,
bioavailability is generally determined by pharmacokinetic measures, i.e., those relating
to the drug absorbed amount and the absorption process speed. These measures may be
obtained from the drug quantification results in biological fluids such as blood or urine,
after single-dose extravascular administration.
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The choice of designs and statistical methods for data analysis are two important
aspects in a bioequivalence study. These two aspects are much related to each other,
once the analysis method depends on the used design. The most used experimental
planning in bioavailability / bioequivalence assays is the crossover one.
Drug effect is the one observed during the period in which it is administered, while
carryover effect is the drug effect that persists after the end of the dosage period. The
existence of carryover effect means that there are different carryover effects in the
sequences of treatment; The non-existence of carryover effect does not necessarily
imply such effects are null, but that if existing, they have the same intensity in both
sequences of treatment.
Usually a complete crossover design is used. With this design, each subject receives all
products with a washout period between each dose administration.
(A). Open-Label
A term used to describe the situation when both the researcher and the participant in a
research study know the treatment the participant is receiving. Open-label is the
opposite of double blind when neither the researcher nor the participant knows what
treatment the participant is receiving.
Keeping group assignment (e.g. to treatment or control) secret from the study
participants or investigators. Blinding is used to protect against the possibility that
knowledge of assignment/drug may affect patient response to treatment, provider
behaviors (performance bias) or outcome assessment (detection bias). Blinding is not
always practical (e.g. when comparing surgery to drug treatment). The importance of
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blinding depends on how objective the outcome measure is; blinding is more important
for less objective outcome measures such as pain or quality of life.
Neither the participants in a trial nor the investigators (outcome assessors) are aware of
which intervention the participants are given. The purpose of blinding the participants
(recipients and providers of care) is to prevent performance bias. The purpose of
blinding the investigators (outcome assessors, who might also be the care providers) is
to protect against detection bias.
Systematic differences in the care provided apart from the intervention being evaluated.
For example, if patients know they are in the control group, they may be more likely to
use other forms of care, patients who know they are in the experimental (intervention)
group may experience placebo effects, and care providers may treat patients differently
according to what group they are in. Blinding of study participants (both the recipients
and providers of care) is used to protect against performance bias.
Systematic differences between comparison groups and how, outcomes are ascertained
diagnosed or verified.
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having the randomization process administered by someone who is not responsible for
recruiting participants; for example, a hospital pharmacy, or a central office. Using
methods of assignment such as date of birth and case record numbers (see quasi-
random allocation) are open to manipulation. Adequate methods of allocation
concealment include: centralized randomization schemes; randomization schemes
controlled by a pharmacy; numbered or coded containers in which capsules from
identical-looking, numbered bottles are administered sequentially; on-site computer
systems, where allocations are in a locked unreadable file; and sequentially numbered
opaque, sealed envelopes.
A study in which patients are allocated at random (by chance alone), to receive one of
several clinical interventions. One of these interventions is the standard of comparison
or control. The control may be a standard practice, a placebo (sugar pill), or no
intervention at all. Someone who takes part in a randomized controlled trial (RCT) is
called a participant or subject. RCTs seek to measure and compare the outcomes after
the participants receive the interventions. Because the outcomes are measured, RCTs
are quantitative studies.
A research design where the subjects will get both the treatments in sequence. Contrast
this with a parallel groups design where some subjects get the first treatment and
different subjects get the second treatment. The crossover design represents a special
situation where there is not a separate comparison group. In effect, each subject serves
as his/her own control. In addition, since the same subject receives both treatments,
there is no possibility of covariate imbalance. Ideally, in a crossover design, a subject is
randomly assigned to a specific treatment order. Some subjects will receive the
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standard therapy first, followed by the new therapy (AB). Others will receive the new
therapy first, followed by the standard therapy (BA).
A research design, where we apply the treatment and the control (or the two treatments)
simultaneously to two separate groups of subjects. Contrast this with a crossover design
where each subject gets the treatment and then the control (or the control and then the
treatment) in sequence.
A research design, where subjects are assessed at a single time in their lives. A cross
sectional study is fast and can study a large number of patients at little cost or effort.
Also, we dont have to worry about patients dropping out during the course of the
study. This study is efficient at identifying association, but may have trouble deciding
cause and effect. With data at only one time point, we do not know whether the chicken
or the egg came first.
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This is a conventional not-replicated design with two formulations, two periods, two
sequences that may be represented as follows:
Randomization for a 2x2 crossover study may be carried out through tables of random
numbers or randomization procedures implemented by statistical software.
The same test and reference formulation batches shall be used for this design for
replicated administration. The periods shall be sufficiently spaced (washout) to assure
non-existence of carryover effects. More commonly used replicated crossover designs
to compare two formulations are:
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1 T T
2 R R
3 R T
4 T R
Or
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In the Code of Federal Regulations (CFR), the U.S. Food and Drug Administration
regulations state that a CRO is "a person [i.e., a legal person, which may be a
corporation] that assumes, as an independent contractor with the sponsor, one or more
of the obligations of a sponsor, e.g., design of a protocol, selection or monitoring of
investigations, evaluation of reports, and preparation of materials to be submitted to the
Food and Drug Administration." [21 CFR 312.3(b)]
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The CRO industry developed mostly in the late 1990s when pharmaceutical R&D
efforts became more complex and competition in rapidly-growing therapeutic areas
increased. Particularly over the last few years, this forced the pharmaceutical industry
to utilize downsizing strategies more to concentrate resources on core skills. As
industry margins come under increasing pressure, companies could begin outsourcing
aspects of their development, manufacturing or marketing processes so as to
concentrate on their core specialties.
External cost pressures have acted as a major driver for the pharmaceutical outsourcing
market. At bottom, the outsourcing market has developed in response to the downward
and upward cost pressures exerted on pharmaceutical manufacturers profit margins.
Given that such pressures are likely to increase in the future, CROs will become more
and more important strategic partners for pharmaceutical companies. It is, therefore, in
the latters interest to consider probable developments in the CRO market and its major
players.
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Sponsor can convert the fixed costs of maintaining the personnel, expertise and
facilities like data management necessary for clinical trial management into
variable costs
Non-availability of services in-house
Knowledge of regulatory affairs in a particular country of interest
Increased complexity of clinical trials
Necessity for medical and clinical knowledge in specific therapeutic areas or
indications
Increased amount of data required from clinical trials
Multinational and multi-center nature of current clinical trials
Large requirement of patient populations
Regionalized diseases
Industry analysers IMS Health and BCC Research estimate that the global
pharmaceutical market will grow at about a 5% rate in 2009 to over $820 billionand be
worth over $1 trillion by 2013. Pharmaceutical and biotechnology companies in the US
spent approximately $59 billion on R&D in 2007, which equates to roughly 18% of
their sales and is a 5% increase from the previous year.
A significant portion of R&D budgets are used for the outsourcing services offered by
the CRO industry, approximately $15 billion in 2007. This figure is expected to grow at
15% over the next seven years and should increase further with the broadening of the
spectrum of services outsourced to cover the entire value chain. As outsourced services
in developing countries such as China and India move up the value chain to cover
phase 1/2 trials, the total contracts value may go up to $20 billion by 2010. Further,
certain therapeutic areas within pharmaceutical development are slated for an even
greater growth curve, namely the oncology class, expected to see continued growth of
upwards of 21% over the next few years due to the large target market, strong unmet
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medical need, and overwhelming number of drugs currently in development (667 for
cancer vs. 252 for CNS disorders, 206 for cadiovascular disorders, and 186 for
infections). [13]
Investigator is a person responsible for the conduct of the clinical trial at a trial site. If a
trial is conducted by a team of individuals at a trial site, the investigator is the
responsible leader of the team and may be called the principal investigator (PI).
The principal investigator (PI) is charged to conduct objective research that generates
independent, high quality, and reproducible results. The Principal Investigator is
responsible for the management and integrity of the design, conduct, and reporting of
the research project and for managing, monitoring, and ensuring the integrity of any
collaborative relationships. Additionally, the principal investigator is responsible for
the direction and oversight of compliance, financial, personnel, and other related
aspects of the research project to assure research is conducted in accordance with
applicable regulatory requirements and ethics committee. It is a responsibility of the
principal investigator to assure that the rights, safety and well-being of the research
subjects are guaranteed, correctly and promptly.
A project manager is the person accountable for accomplishing the stated project
objectives. The project manager's role in a nutshell is the overall responsibility for the
successful planning, execution, monitoring, control and closure of a project. Key
project management responsibilities include creating clear and attainable project
objectives, building the project requirements, and managing the triple constraint for
projects, which are cost, time, and quality (also known as scope).
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Quality assurance (QA) refers to a program for the systematic monitoring and
evaluation of the various aspects of a project, service, or facility to ensure that
standards of quality are being met.
They liaise with other managers and staff throughout the organisation to ensure that the
QA system is functioning properly. Where appropriate, the quality manager advises on
changes and their implementation and provides training, tools and techniques to enable
others to achieve quality.
Clinical Research Coordinator (CRC) is the individual working for the investigator or a
university/academic institution who handles most of the administrative responsibilities
of a clinical trial/clinical study, is the liaison between the investigators site and the
sponsor, and reviews all data and records related to the clinical trial/clinical study.
Clinical Research Coordinator (CRC) is responsible for the entire conduct of the study,
using good clinical practice (GCP) under the auspices of the Principal Investigator (PI).
The main function of a clinical research associate is to monitor clinical trials. A clinical
research associate ensures compliance with the clinical trial protocol, checks clinical
site activities, makes on-site visits, reviews Case Report Forms (CRFs) and
communicates with clinical research investigators.
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A medical writer, working with doctors, scientists, and other subject matter experts,
creates documents that effectively and clearly describe research results, product use,
and other medical information. The medical writer also makes sure that documents
comply with regulatory requirement, ethics committee, Good Clinical Practice (GCP)
and other applicable guidelines in terms of content, format and structure. Their main
responsibility in a research team is the preparation of study protocol and clinical study
reports.
(H) Pharmacist
Pharmacists are healthcare professionals who practice the science of pharmacy. Their
main responsibility is receipt and accountability of investigational product.
(I) Phlebotomist
(J) Nurse
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(K) Custodian
Delayed release
Sustained release
Mixed immediate and sustained release
Mixed delayed and sustained release
Mixed immediate and delayed release
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Produce plasma levels which lie within the therapeutic range (where
appropriate) for the proposed dosing intervals at steady state.
If all of the above conditions are not met but the applicant considers the formulation to
be acceptable, justification to this effect should be provided.
I. Study Parameters
Bioavailability data should be obtained for all modified release drug products although
the type of studies required and the pharmacokinetic parameters which should be
evaluated may differ depending on the active ingredient involved. Factors to be
considered include whether or not the formulation represents the first market entry of
the drug substance, and the extent of accumulation of the drug after repeated dosing.
If the formulation is the first market entry of the drug substance, the products
pharmacokinetic parameters should be determined. If the formulation is a second or
subsequent market entry then comparative bioavailability studies using an appropriate
reference product should be performed.
Study design will be single dose or single and multiple dose based on the modified
release products that are likely to accumulate or unlikely to accumulate both in the
fasted and non-fasting state. If the effect of food on the reference product is not known
(or it is known that food affects its absorption), two separate two-way cross-over
studies, one in the fasted state and the other in the fed state, may be carried out. If it is
known with certainty (e.g. from published data) that the reference product is not
affected by food, then a three-way cross-over study may be appropriate with:
This section outlines the requirements for modified release formulations which are used
at a dose interval that is not likely to lead to accumulation in the body (AUC 0-T/AUC0-
0.8).
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When the modified release product is the first market entry of that type of dosage form,
the reference product should normally be the innovators immediate-release
formulation. The comparison should be between a single dose of the modified release
formulation and doses of the immediate-release formulation which it is intended to
replace. The latter must be administered according to the established dosing regimen.
When the modified release product is the second or subsequent entry on the market,
comparison should be with the reference modified release product for which
bioequivalence is claimed.
Studies should be performed with single dose administration in the fasting state as well
as following an appropriate meal at a specified time. The following pharmacokinetic
parameters should be calculated from plasma (or relevant biological matrix)
concentrations of the drug and/or major metabolite(s): AUC0-T, AUC0-t, AUC0-, Cmax
(where the comparison is with an existing modified release product), and Kel.
The 90% confidence interval calculated using log-transformed data for the ratios (Test:
Reference) of the geometric mean AUC (for both AUC0-T and AUC0-t) and Cmax (where
the comparison is with an existing modified release product) should generally be within
the range 80 to 125% both in the fasting state and following the administration of an
appropriate meal at a specified time before taking the drug.
The pharmacokinetic parameters should support the claimed dose delivery attributes of
the modified-release dosage form.
This section outlines the requirements for modified release formulations that are used at
dose intervals that are likely to lead to accumulation (AUC0-T/AUC0- < 0.8).
When a modified release product is the first market entry of the modified release type,
the reference formulation is normally the innovators immediate-release formulation.
Both a single dose and steady state doses of the modified release formulation should be
compared with doses of the immediate-release formulation which it is intended to
replace. The immediate-release product should be administered according to the
conventional dosing regimen.
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Studies should be performed with single dose administration in the fasting state as well
as following an appropriate meal. In addition, studies are required at steady state. The
following pharmacokinetic parameters should be calculated from single dose studies:
AUC0-T, AUC0-t, AUC0-, Cmax (where the comparison is with an existing modified
release product), and Kel. The following parameters should be calculated from steady
state studies: AUC0-T(ss), Cmax, Cmin, Cpd and degree of fluctuation.
When the modified release product is the second or subsequent modified release entry,
single dose and steady state comparisons should normally be made with the reference
modified release product for which bioequivalence is claimed.
The 90% confidence interval for the ratio of geometric means (Test: Reference drug) of
AUC (for both AUC0-T and AUC0-t) and Cmax (where the comparison is with an existing
modified release product) determined using log-transformed data should generally be
within the range 80 to 125% when the products are compared after single dose
administration in both the fasting state and the fed state.
The 90% confidence interval for the ratio of geometric means (Test: Reference drug)
for AUC0-T(ss), Cmax and Cmin determined using log-transformed data should generally
be within the range 80 to 125% when the formulations are compared at steady state.
The pharmacokinetic parameters should support the claimed attributes of the modified-
release dosage form.
When these studies do not show bioequivalence, comparative efficacy and safety data
may be required for the new product. [14]
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It is the objective of regulatory guidance to define, for products with a systemic effect,
when bioavailability or bioequivalence studies are necessary and to formulate
requirements for their design, conduct, and evaluation. The possibility of using in vitro
instead of invivo studies with pharmacokinetic end points is also envisaged.
Guidance documents are administrative instruments not having force of law and, as
such, allow for flexibility in approach. Alternate approaches to the principles and
practices described in document may be acceptable provided they are supported by
adequate scientific justification. Alternate approaches should be discussed in advance
with the relevant program area to avoid the possible finding that applicable statutory or
regulatory requirements are met.
The requirements for conduct of Bioavailability and Bioequivalence studies vary from
country to country. These changes are reflected in the guidelines issued by the different
regulatory authorities across the world.
Some of the Regulatory Authorities which have issued guidelines for the conduct of
Bioavailability and Bioequivalence studies are as follows:
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s [15]
regulatory
authorities own
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3.11.1 Epidemology
Globally diabetes affects 246 million people and is expected to affect 366 million by
2030. At least 7 million new cases are reported annually. India accounts for largest
diabetic population with 41 million patients, mounting to 6 percent of the adult
population.
Type 1 diabetes mellitus results from autoimmune destruction of the cells of the
pancreas. Type 1 diabetes mellitus develops in childhood or early adulthood, although
some latent forms do occur. Type 1 DM accounts for up to 10% of all cases of DM and
is likely initiated by the exposure of a genetically susceptible individual to an
environmental agent.
Among various types of diabetes mellitus, type 2 diabetes mellitus is the commonest.
The prevalence of type 2 DM is increasing. Type 2 DM accounts for as much as 90% of
all cases of DM. Type 2 Diabetes Mellitus is characterized by insulin resistance and at
least initially, a relative lack of insulin secretion. Most individuals with type 2 diabetes
exhibit abdominal obesity which itself causes insulin resistance. Multiple risk factors
for the development of type 2 DM have been identified, including family history (i.e.,
parents or siblings with diabetes); obesity (i.e., 20% over ideal body weight, or body
mass index [BMI] 25 kg/m2); habitual physical inactivity; race or ethnicity;
previously identified impaired glucose tolerance or impaired fasting glucose;
hypertension (140/90 mm Hg in adults); high density lipoprotein (HDL) cholesterol
35 mg/dL and/or a triglyceride level 250 mg/dL; history of gestational diabetes
mellitus or delivery of a baby weighing >9 pounds; history of vascular disease; and
polycystic ovary disease. While the prevalence of type 2 DM increases with age, the
disorder is increasingly being recognized in adolescence. Much of the rise in adolescent
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3.11.2 Classification
[17]
Random blood sugar (RBS): 200 mg/dL (Normal Value: 60-150 mg/dl)
Fasting blood sugar (FBS): 126 mg/dL (Normal Value: 60-110 mg/dl)
Post Prandial blood sugar (PPBS): 200 mg/dL (Normal Value: 90-140 mg/dl)
Oral glucose tolerance test (OGTT, 75 g glucose): 200 mg/dL
(Normal Value: <140 mg/dl)
Glycosylated hemoglobin (Hb A1C): >7% (Normal Value: <7%)
C-peptides
Urine analysis
Criteria for screening diabetes in asymptomatic, undiagnosed individuals: > 45
years, Younger people (obese, first degree relative with DM, high risk ethnic
population, baby wt> 9 lb with GDM, HTN, HDL35 mg/dl, TG250 mg/dL)
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IGT: Impaired Glucose Tolerance, FPG: Fasting Plasma Glucose, OGTT: Oral Glucose
Tolerance Test
Metformin should be included in the therapy for all type 2 DM patients, if tolerated and
not contraindicated, as it is the only oral antihyperglycemic medication proven to
reduce the risk of total mortality and cardiovascular death, according to the United
Kingdom Prospective Diabetes Study.
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Knowledge of the patients quantitative and qualitative meal patterns, activity levels,
pharmacokinetics of insulin preparations, and pharmacology of oral antihyperglycemic
agents are essential to individualize the treatment plan and optimize blood glucose
control while minimizing risks for hypoglycemia and other adverse effects of
pharmacologic therapies.
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1.Sulfonylureas:
2. Biguanides: Metformin
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3.11.4.2 Biguanides
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3.11.4. Thiazolidinediones
3.11.4.5 Meglitinides
3.11.4.6 Insulin
Insulin is mainstay in the treatment of diabetes mellitus (type 1 and type 2 also). Type 1
treatment necessitates insulin therapy. Currently, the basal-bolus insulin therapy or
pump therapy in motivated individuals often leads to successful glycemic outcomes.
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Basal-bolus therapy often includes a basal insulin for fasting and post-absorptive
control, and rapid acting bolus insulin for mealtime coverage. Therapeutically, use of
basal-bolus therapy in type 2 DM is increasing.
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(C) Site
(D) Route
Subcutaneous.
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[20]
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3.12.1 Summary
3.12.2 Introduction
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3.12.4 Storage
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3.12.6 Pharmacokinetics
3.12.6.1 Absorption
At steady state, the AUC and Cmax are less than dose proportional for Metformin
hydrochloride sustained release within the range of 500 mg to 2000 mg administered
once daily. Peak plasma levels are approximately 0.6, 1.1, 1.4, and 1.8 g/mL for 500,
1000, 1500, and 2000 mg once-daily doses, respectively. The extent of metformin
absorption (as measured by AUC) from Metformin hydrochloride sustained release at a
2000 mg once-daily dose is similar to the same total daily dose administered as
Metformin hydrochloride tablets 1000 mg twice daily. After repeated administration of
Metformin hydrochloride sustained release, metformin did not accumulate in plasma.
Although the extent of metformin absorption (as measured by AUC) from the
Metformin hydrochloride sustained release tablet increased by approximately 50%
when given with food, there was no effect of food on Cmax and Tmax of metformin.
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Both high and low fat meals had the same effect on the pharmacokinetics of Metformin
hydrochloride sustained release.
3.12.6.2 Distribution
The apparent volume of distribution (V/F) of metformin following single oral doses of
Metformin hydrochloride 850 mg averaged 654 358 L. Metformin is negligibly
bound to plasma proteins, in contrast to sulfonylureas, which are more than 90%
protein bound. Metformin partitions into erythrocytes, most likely as a function of time.
At usual clinical doses and dosing schedules of Metformin hydrochloride, steady state
plasma concentrations of metformin are reached within 24 to 48 hours and are
generally <1 g/mL. During controlled clinical trials of Metformin hydrochloride,
maximum metformin plasma levels did not exceed 5 g/mL, even at maximum doses
3.12.6.3 Metabolism
3.12.6.4 Elimination
Renal clearance is approximately 3.5 times greater than creatinine clearance, which
indicates that tubular secretion is the major route of metformin elimination. Following
oral administration, approximately 90% of the absorbed drug is eliminated via the renal
route within the first 24 hours, with a plasma elimination half-life of approximately 6.2
hours. In blood, the elimination half-life is approximately 17.6 hours, suggesting that
the erythrocyte mass may be a compartment of distribution.
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Diarrhea
Nausea
Upset Stomach
Abdominal Bloating
Gas
Loss of Appetite
These side effects generally go away after you take the medicine for a while. Taking
your medicine with meals can help reduce these side effects.
Adverse reactions reported in greater than 5% of the Metformin patients, and that were
more common are Diarrhea, Nausea/vomiting, Flatulence, Asthenia, Indigestion,
Abdominal discomfort, Headache.
Other Adverse reactions less commonly reported were abnormal stools, hypoglycemia,
myalgia, lightheaded, dyspnea, nail disorder, rash, sweating increased, taste disorder,
chest discomfort, chills, flu syndrome, flushing, palpitation, upper respiratory infection
and taste disturbance.
The single-dose nature of this study and the lack of correlation between glyburide
blood levels and pharmacodynamic effects make the clinical significance of this
interaction uncertain.
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3.12.10.5 OtherCertain drugs tend to produce hyperglycemia and may lead to loss of
glycemic control. These drugs include the thiazides and other diuretics, corticosteroids,
phenothiazines, thyroid products, estrogens, oral contraceptives, phenytoin, nicotinic
acid, sympathomimetics, calcium channel blocking drugs, and isoniazid. When such
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Metformin is negligibly bound to plasma proteins and is, therefore, less likely to
interact with highly protein-bound drugs such as salicylates, sulfonamides,
chloramphenicol, and probenecid, as compared to the sulfonylureas, which are
extensively bound to serum proteins.
Lactic acidosis is a rare, but serious, metabolic complication that can occur due to
metformin accumulation during treatment with metformin or metformin SR; when it
occurs, it is fatal in approximately 50% of cases. Lactic acidosis may also occur in
association with a number of pathophysiologic conditions, including diabetes mellitus,
and whenever there is significant tissue hypoperfusion and hypoxemia.
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metformin, such as cationic drugs that are eliminated by renal tubular secretion should
be used with caution.
3.12.12.7 Impaired hepatic function Since impaired hepatic function has been
associated with some cases of lactic acidosis, metformin and metformin SR should
generally be avoided in patients with clinical or laboratory evidence of hepatic disease.
3.12.13 Contraindications
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A 24-week, double-blind, randomized study of Diabeta SR, taken once daily with the
evening meal, and metformin hydrochloride tablets, taken twice daily (with breakfast
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and evening meal), was conducted in patients with type 2 diabetes who had been
treated with metformin hydrochloride 500 mg tablets twice daily for at least 8 weeks
prior to study entry. The metformin hydrochloride tablets dose had not necessarily been
titrated to achieve a specific level of glycemic control prior to study entry. Patients
qualified for the study if HbA1c was 8.5% and FPG was 200 mg/dL.After 12 weeks
of treatment, there was an increase in mean HbA1c in all groups; in the Diabeta SR
1000 mg group, the increase from baseline of 0.23% was statistically significant. [24]
Metformin absorption is relatively slow and may extend over about 6 hours. Animal
14
studies with metformin, labelled with C have shown that the drug is neither
concentrated by liver cells nor is it excreted in the bile; it is concentrated in the
intestinal mucosa and salivary glands.
It has been shown that, following a 2 g dose of metformin, the blood level remains
under 10 mcg/mL even at the peak, occurring 2 hours after absorption. During the
experiments, metformin was shown to be devoid of any notable action in the body,
apart from its specific metabolic activity.
In the healthy animal, metformin lowers blood sugar only at a nearly lethal dose.
Different animal species are of unequal sensitivity. On the other hand, the animal with
experimental diabetes is sensitive to a much lower dosage, providing some insulin is
still secreted. The antihyperglycemic action of metformin is probably mediated through
insulin: Metformin improves the K co-efficient of glucose assimulation. Metformin
improves the co-efficient of insulin efficiency. In the obese diabetic with
hyperinsulinemia, metformin is reported to normalize insulin output. This normalizing
effect is concurrent to that of glycemia. Metformin has little effect on liver glycogen of
the healthy animal. In low and average doses, no change occurs. In high doses nearing
lethal levels, liver glycogen decreases. This lowering precedes the fall in blood sugar.
This reaction represents a defense mechanism tending to mobilize body reserves in
order to combat hypoglycemia.
In the diabetic animal with a low liver glycogen reserve, the opposite occurs and
metformin builds up glycogen stores of the liver. In vitro, on muscular tissue isolated in
Warburgs apparatus, metformin increases glucose uptake by the muscle. This action
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Recent findings appear to indicate that most of the metabolic effects of the biguanides
are exerted through a single mechanism, namely inhibition of fatty acid oxidation and
of acetyl-CoA generation.
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Long-term carcinogenicity studies have been performed in rats (dosing duration of 104
weeks) and mice (dosing duration of 91 weeks) at doses up to and including 900
mg/kg/day and 1500 mg/kg/day, respectively.
These doses are both approximately four times the maximum recommended human
daily dose of 2000 mg based on body surface area comparisons. No evidence of
carcinogenicity with metformin was found in either male or female mice. Similarly,
there was no tumorigenic potential observed with metformin in male rats. There was,
however, an increased incidence of benign stromal uterine polyps in female rats treated
with 900 mg/kg/day.
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Recent information strongly suggests that abnormal blood glucose levels during
pregnancy are associated with a higher incidence of congenital abnormalities. Most
experts recommend that insulin be used during pregnancy to maintain blood glucose
levels as close to normal as possible. Because animal reproduction studies are not
always predictive of human response, DIBETA SR should not be used during
pregnancy unless clearly needed.
There are no adequate and well-controlled studies in pregnant women with DIBETA
SR. Metformin were not teratogenic in rats and rabbits at doses up to 600 mg/kg/day.
This represents an exposure of about two and six times the maximum recommended
human daily dose of 2000 mg based on body surface area comparisons for rats and
rabbits, respectively. Determination of fetal concentrations demonstrated a partial
placental barrier to metformin. [25]
4. Methodology
This study was conducted at Auriga Research Ltd (A Delhi based CRO). Auriga
Research Ltd. is the sister organization of Arbro Pharmaceuticals Ltd, New Delhi and
both the organizations are located in Kirti Nagar Industrial Area, New Delhi. The
various facilities offered by Auriga Research Ltd. are as follows:
I. Clinical Facility
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LC-MS/MS lab.
HPLC and GC lab.
ICP-MS lab.
GC-MS lab.
Cold room and Deep freezer for sample storage (at -800C).
Sample preparation area.
Wet lab equipped with Refrigerated centrifuges, Airshaft free micro and
analytical balance areas, Fume hoods.
Dark lab.
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Metformin Hydrochloride (SR) 500 mg tablet was the Test Product and Dibeta SR
tablet [containing Metformin Hydrochloride (SR) 500 mg] was used as the Reference
standard.
Metformin Hydrochloride (SR) 500 mg tablet was the Test Product and Dibeta SR
tablet [containing Metformin Hydrochloride (SR) 500 mg] was the Reference standard.
The sponsor had supplied sufficient quantities of the study formulations for
conduct of the study & for retention purpose.
The drug products along with Certificate of Analysis (COA) was received by
the Pharmacist or Principal Investigator or designee.
The COA had product description/appearance, identification, in vitro
dissolution data and assay as per specifications/pharmacopoeia
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Reference products was supplied in the original manufacturers packing and the
test products was supplied in an appropriate package deemed to maintain the
integrity of the products
Records were made of the receipt and dispensing of clinical supplies to provide
a complete accountability of all supplies
The supplies were stored at controlled temperature or as per label instructions in
Pharmacy accessible only to the Pharmacist or authorized personnel
Batch number, manufacturing date, expiry date and physical description for
both products was included in the final report.
At the conclusion of the study, the unused supplies were retained in accordance
to applicable regulatory requirements regarding retention of reserve sample.
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DISPENSING
Period I
Dibeta SR 149+01*
AF-9010 06+01** 142
500 mg (R) =150 tablets
Period II
Dibeta SR 141+01*
AF-9010 06+01** 134
500 mg (R) =142 tablets
RETENTION
The following information was given on the dosing label prepared for the study:
Study No.
Period No.
Subject No.
Randomization Code
Date of Dosing
Dated initials of Pharmacist
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4.2.4 Labeling
Whether a subject receives test or reference products during the study was
determined according to randomization schedule generated and authorized by
PK Scientist.
The Clinical Investigator and the personnel involved in dispensing of study
drugs were accountable for ensuring compliance to randomization schedule.
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Period I Period II
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4.3.4 Dosing
After a supervised overnight fast, subjects had received a single oral dose of the
assigned formulation, with 240 ml of 20% solution of glucose, according to the
randomization schedule generated and authorized by Biostatistician. Mouth check was
done by the study personnel doing dosing in the presence of other study personnel
responsible for dosing supervision. To prepare 20% of glucose, 48 gram of
commercially available glucose powder was dissolved in 240 mL of water.
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Recruited Volunteers
N=12
Period I Period II
N=12 N=12
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4.3.8.2 Prohibitions
The subjects were prohibited from smoking or consuming tobacco containing products,
xanthine containing products and alcohol during their stay in the clinical facility.
4.3.9 Blinding
Auriga Research Limited had make no changes in the appearance of test or reference
product. While dosing, the treatments were labeled as mentioned under section 4.2.4.
The pharmacist or a person designated by the principal investigator had dispensed and
label the product as per randomization code.
Equal allocation of subjects to each sequence was ensured. Study personnel involved in
the sample analysis were kept blinded from the randomization code during the entire
study. Persons involved in dispensing of study drug and in verifying the dispensing
activity were accountable for ensuring compliance with the Randomization schedule.
The randomization schedule used in this study was as follows:
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1 B RT R T
2 A TR T R
3 A TR T R
4 B RT R T
5 B RT R T
6 A TR T R
7 A TR T R
8 B RT R T
9 A TR T R
10 B RT R T
11 A TR T R
12 B RT R T
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The blood samples were collected and transferred into pre-labeled (mentioning Study
number, Subject number, Period and sampling time point) K2EDTA sample collection
tube. If for any reason the indwelling cannula / scalp vein set is blocked or must be
removed for practical reasons, direct venipuncture was done. For each subject the total
number of blood samples drawn during the study was 28 and the total volume of blood
drawn including 10 ml for screening, 11 ml of blood which is discarded (i.e., 0.5 ml of
blood is discarded before each sample is drawn to remove the heparin which is in the
indwelling cannula / scalp vein set), had not exceed 111 ml for the entire study.
The choice of the blood sampling time schedule was intended to cover the expected
time of the peak plasma concentrations of metformin, which was at around 7 (4-8) hour
(Drugs @ FDA, 2008). It was also extended for sufficient time to be able to obtain a
reliable estimate of the extent of absorption. The last sample collection was at 24.00
hours post-dose, which was more than 3 times the maximum reported half-life of
metformin 6.2 hours, and was cover at least 80% of the AUC0-.
Phlebotomy
6.0 7.0 8.0 9.0 10.0 12.0
Schedule (hrs)
18.0 24.0
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The samples were transported to the Arbro Pharmaceutical Ltd analytical section
within 12 hr of sample separation and stored at (80 10)0C or colder till the time of
analysis.
4.3.14 Criteria for Recording & Reporting Blood Sample Deviation
Blood samples were collected at times specified under phlebotomy schedule. The
actual time of collection of each blood sample was the time at which sample collection
end and the hour and minutes on clock display at that time was recorded on the
appropriate raw data forms. A window period of 2 minutes from the scheduled time
point is allowed for the in-house blood draws. All deviations outside the allowed range
as above were documented as protocol deviations. In all such instances appropriate
time corrections for the actual time of sample collection, was incorporated at the time
of data analysis.
4.3.15 Study Meals
Standardized meals (Annexure-VIII) prepared by dietician was served to subjects. An
overnight fasting of at least 10 hrs was ensured prior to dosing and fasting state was
continued until 4 hrs post dose. Lunch, snacks and dinner were provided at 4, 9 & 13
hrs post dose respectively. Water was restricted from 1 hr Predose to 2 hrs post dose.
Subsequently water was provided ad libitum. 60 mL of 20% glucose solution was
provided at the interval of 15 minutes till 4 hrs of post dose, and whenever required
after blood glucose monitoring or safety reason.
4.3.16 Housing
From at least 12 hours prior to drug administration until after 24 hours. post dose
4.3.17 Washout Period
At least 07 days between treatments.
4.3.18 Activity Levels
Subjects were seated upright for the first 2 hours following drug administration.
However, should medical events occur at any time, subjects may be placed in an
appropriate position; Subjects were prohibited from any strenuous or athletic activity
during housing.
4.4 Clinical Assessment
4.4.1 Subject Screening
Subject Screening was done 21 days prior to the start of the study. Medical histories
and demographic data including name, sex, age, race, body weight (kg), and height
(cm), and smoking habits were recorded. Each subject has to undergo a complete
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general physical examination and laboratory tests as mentioned in the next section
4.4.2. Only medically healthy subjects with clinically acceptable laboratory profiles,
chest X-ray (performed within 6 months prior to study start) and ECG (performed
within 21 days prior to study start) were enrolled into the study.
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Indian GCP and the principles enunciated in the Declaration of Helsinki (Ethical
Principles for Medical Research Involving Human Subjects, revised by 59th WMA
General Assembly, Seoul, October 2008).
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difference. The above analysis was done using the WinNonlin Version 5.2 version.
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4.10 Archives
All raw data generated in connection with this study, together with the original copy of
the final report, was retained in the archival area of Auriga Research Limited for at
least 2 yrs after the expiration date of the batch as per CDSCO.
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3) Screening
4) Study Procedures
Period I
ICD Presentation
Provide ample time to volunteers to understand the ICD
Obtain the consent of volunteers on ICF.
Check in of the volunteers
Vital measurements & Medical examination
Dinner as per study meal menu
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Period II
Same as Period I
Discharge with remaining 70% compensation
Hippocrates Independent Ethics Committee (HIEC), New Delhi has given the approval
to conduct this study and allowed to enroll twelve subjects for this particular study
182
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based on ethical reason. All the enrolled twelve subjects were of age between 18 and
45 years, weight within 15% of the ideal Height-Weight chart of Life Insurance
Corporation (LIC) of India (see Annexure V). Table 19 &20 describes the demographic
characters like gender, age, height, weight and diet of the enrolled volunteers.
Non- 24-10-09
4 Male 36 169 64 22.40 Veg
Smoker
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Veg Smoker
N 12 12 12
Screening examination was conducted within 21 days prior to study. During screening,
medical history of each volunteer was taken. Laboratory tests (including Hematology,
Biochemistry Serology and Urine examination) were also performed for each
volunteer. After passing the screening examination, healthy volunteers were enrolled in
the study.
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1 2 3 4 5 6 7 8 9 10 11 12
HAEMOGLOBIN (Hb) 13 - 17 13.0 13.7 13.3 13 13.5 13.3 13.1 13 15.7 14.9 13.6 13.3
TLC 4 - 10 5.3 6.1 8.8 4.5 8.8 7.1 9.5 9.4 8.6 8.6 6.1 9.1
NEUTROPHIL 40 - 80 65 57 70 40 70 66 50 62 57 62 72 61
LYMPHOCYTE 20 - 40 28 35 26 40 24 28 39 33 38 30 22 30
EOSINOPHIL 01 - 06 4 5 1 3 2 1 3 1 2 3 2 6
MONOCYTE 02 - 10 3 3 3 2 4 5 3 4 3 5 4 3
BASOPHIL <01 02 0 0 0 0 0 0 0 0 0 0 0 0
PLATELET COUNT 150 - 410 193 380 202 273 226 156 210 157 219 156 165 178
CREATININE 0.9 - 1.3 1.0 1.1 0.9 0.9 0.9 1 0.9 1.3 1.1 1 1.1 1.3
TOTAL BILIRUBIN 0.0 1.0 0.8 0.3 0.4 0.5 0.6 0.4 0.7 0.3 0.4 0.5 0.3 0.3
S.G.O.T(AST) 15 - 37 30 29 26 26 37 28 18 35 23 20 23 30
S G.P.T(ALT) 30 - 63 59 43 47 39 56 58 35 52 40 42 63 41
TOTAL PROTEIN 6.4 - 8.2 7.6 7.9 8 7.5 7.9 7.3 6.4 8.2 8.1 6.7 8 8.1
TOTAL CHOLESTEROL <200 103 182 151 159 195 122 92 161 150 175 115 148
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SEROLOGY
V.D.R.L NR NR NR NR NR NR NR NR NR NR NR NR NR
HBsAg (Qualitative) NR NR NR NR NR NR NR NR NR NR NR NR NR
Out of range parameters (Highlighted) considered as Insignificant or Clinically Insignificant), NR: Non Reactive
Test Name
1 2 3 4 5 6 7 8 9 10 11 12
Physical
Examination
Quantity (ml) 25 25 25 25 25 25 25 25 25 25 25 25 25
Colour N/AP PY PY PY PY PY PY PY PY PY PY PY PY
Appearance N/AP CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR CLEAR
Chemical
Examination
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Microscopic
Examination
WBC(Pus Cells)
/HPF 0-5 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1
Casts N/AP ND ND ND ND ND ND ND ND ND ND ND ND
Crystals N/AP ND ND ND ND ND ND ND ND ND ND ND ND
Epithelial Cells 0-5 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1
Others Specify ND ND ND ND ND ND ND ND ND ND ND ND
SP.Gravity 1.003 - 1.035 1.015 <=1.005 <=1.005 <=1.005 <=1.005 1.010 <=1.005 <=1.005 1.010 <=1.005 <=1.005 <=1.005
90
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Majority of the post-dose samples were collected within 2 minutes from the
scheduled sampling time in all the periods of the study as per SOP. None of the
deviations were observed in blood sample collection.
None
*Reason
The plasma samples were stored at -20C or colder till the completion of the time
period and then transferred to the bioanalytical facility as per the SOP and were stored
at (-8010)C or colder until analysis. None of the deviations were observed in
plasma storage condition.
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Figure 7: Mean plasma concentration vs. time curve of Test and Reference Metformin Hydrochloride (SR) 500mg tablet formulations
600
500
400
300
R
T
200
100
0
0 5 10 15 20 25
Time (hr)
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Figure 8: Mean of Log plasma concentration of Test and Reference Metformin Hydrochloride (SR) 500mg tablet formulations
6.5
6.0
5.5
R
T
5.0
4.5
4.0
0 2 4 6 8 10 12 14 16 18
Time (hr)
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The rate of absorption was determined by Cmax and tmax. The adequacy of the
sampling time was determined by the ratio (AUC0-t / AUC0-) %. The half life of
elimination t1/2 and the rate of elimination Kel were used to further characterize the
pharmacokinetics outcome of this study. The extent of absorption was determined by
using AUC0-t and AUC0- of Metformin hydrochloride SR 500 mg.
AUC0-t, AUC0- and Cmax values of Metformin hydrochloride SR 500 mg were used
for bioequivalence estimations.
Pharmacokinetic parameters like Cmax, tmax, t , AUC0-t, AUC0- and Kel were
calculated using plasma concentration vs. time profile of both investigational products
using WinNonlin Version 5.2. These data were statistically analyzed and the
bioequivalence of test and reference formulations were proved.
Administration of the reference preparation, tablet Dibeta SR produced the area under
plasma concentration time curve (AUC0-t) 5768.22 1425.27ng hr/ml, whereas the
test preparation of tablet Metformin hydrochloride SR produced the area under
plasma concentration time curve (AUC0-t) 5924.45 1290.10ng hr/ml.
When administered as a single dose in the fasting state, the reference preparation,
tablet Dibeta SR, produced the area under plasma concentration time curve up to
infinity (AUC0-) 5960.311439.23ng hr/ml, whereas the test preparation of tablet
Metformin hydrochloride SR produced the area under plasma concentration time
curve (AUC0-) 6159.181354.45ng hr/ml.
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When administered as a single dose in the fasting state, the reference preparation,
tablet Dibeta SR, produced AUC0-t /AUC0- (%) of 96.71 2.00%, whereas the test
preparation of tablet Metformin hydrochloride SR produced AUC0-t /AUC0- (%) of
96.23 1.20
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Table 25: Pharmacokinetic Parameters of Test Product and Reference Standard of the Study Drug
AUC0-t/AUC0-
AUC0-t (ng hr/ml) AUC0- (ng hr/ml) Cmax (ng/ml) tmax (hr) Kel (1/hr) T1/2 (hr) (%)
Subject
No. Test Reference Test Reference Test Reference Test Reference Test Reference Test Reference Test Reference
1 8037.08 6606.88 8498.24 6711.47 827.81 531.61 6.00 4.00 0.22 0.19 3.20 3.65 94.57 98.44
2 4680.02 4654.37 4841.34 4931.23 551.22 540.63 5.00 6.00 0.24 0.18 2.90 3.82 96.67 94.39
3 8128.28 6468.62 8407.66 6561.09 709.38 811.57 4.00 4.00 0.13 0.20 5.20 3.46 96.68 98.59
4 6153.68 5714.70 6375.59 5852.69 619.93 568.32 4.00 5.00 0.17 0.18 4.18 3.80 96.52 97.64
5 6995.02 4111.06 7163.32 4211.99 665.41 533.14 5.00 4.00 0.20 0.21 3.42 3.25 97.65 97.60
6 5464.39 5983.10 5705.61 6509.81 697.61 636.21 7.00 4.00 0.24 0.16 2.85 4.21 95.77 91.91
7 5595.07 6487.58 5926.66 6746.39 552.37 520.93 6.00 8.00 0.20 0.17 3.54 4.16 94.40 96.16
8 5479.86 2699.60 5641.54 2798.27 521.69 503.15 4.00 3.00 0.17 0.18 4.02 3.89 97.13 96.47
9 4315.02 5383.26 4418.55 5495.35 401.81 587.10 3.00 6.00 0.18 0.18 3.94 3.80 97.66 97.96
10 4887.00 8212.56 5179.73 8302.02 653.97 915.92 6.00 5.00 0.19 0.23 3.72 3.06 94.35 98.92
11 4695.21 6385.60 4831.00 6613.97 551.74 674.38 4.00 3.00 0.17 0.17 4.07 4.13 97.19 96.55
12 6662.79 6511.29 6920.94 6789.42 701.22 630.25 9.00 6.00 0.16 0.15 4.39 4.66 96.27 95.90
Mean 5924.45 5768.22 6159.18 5960.31 621.18 621.10 5.25 4.83 0.19 0.18 3.79 3.82 96.24 96.71
SD 1290.10 1421.27 1354.45 1439.23 112.10 126.66 1.66 1.47 0.03 0.02 0.67 0.44 1.21 2.00
CV(%) 21.78 24.64 21.99 24.15 18.05 20.39 31.59 30.35 17.43 11.86 17.60 11.55 1.26 2.07
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The various mean pharmacokinetic parameters estimated for both the Reference and
Test Product for Metformin Hydrochloride SR 500 mg are described in tables given
below:
AUC0-t (ng
5924.45 1290.10 5768.22 1425.27
hr/ml)
AUC0-(ng
6159.181354.45 5960.311439.23
hr/ml)
AUC0-t/AUC0-
96.23 1.20 96.71 2.00
(%)
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As shown in the Table given below, the 90% confidence intervals were constructed
for the ratios of the geometric least square mean of ln-transformed pharmacokinetic
parameters Cmax, AUC0-t and AUC0- for test and reference formulations.
Bioequivalence was concluded, as the 90% confidence intervals for Cmax, AUC0-t and
AUC0- fall within the acceptable range of 80125%.
The 90% confidence interval for ln-transformed Cmax was 89.13%-112.44%. The
90% confidence interval for ln-transformed AUC0-t was 87.460%-123.850%.The
90% confidence interval for ln-transformed AUC0- was 88.290%-123.870%.
The Mean T/R ratio (%) for ln-transformed Cmax, AUC0-t and AUC0- were
100.110%, 104.080% and 104.580% respectively. Table given below describes the
Geometric mean value; 90% confidence Interval and T/R ratio (%) of the ln-
transformed parameters.
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Table 29: Summary of ANOVA Results obtained from Test and Reference
product of the Study Drug
Log Transformed Cmax Analysis-ANOVA
Source DF S.S M.S.S F stat P value
Sequence 1 0.008 0.008 0.110 0.753
Subject (Seq) 9 0.645 0.072 4.070 0.024
Treatment 1 0.007 0.007 0.410 0.537
Period 1 0.062 0.062 3.500 0.094
Error 9 0.159 0.018 -- --
Total 21 0.880 0.166 8.090 1.408
Log Transformed AUC0-t Analysis-ANOVA
Source DF S.S M.S.S F stat P value
Sequence 1 0.002 0.002 0.040 0.855
Subject (Seq) 9 0.418 0.046 3.040 0.057
Treatment 1 0.005 0.005 0.300 0.598
Period 1 0.002 0.002 0.110 0.751
Error 9 0.138 0.015 -- --
Total 21 0.563 0.070 3.490 2.262
Log Transformed AUC0- Analysis-ANOVA
Source DF S.S M.S.S F stat P value
Sequence 1 0.001 0.001 0.030 0.867
Subject (Seq) 9 0.412 0.046 3.110 0.053
Treatment 1 0.005 0.005 0.310 0.590
Period 1 0.001 0.001 0.050 0.822
Error 9 0.133 0.015 -- --
Total 21.00 0.552 0.067 3.500 2.332
DF = Degree of freedom: S.S = Sum of Squares: M.S.S = Mean Sum of square,
F = M.S.S/Error M.S.S, P = Probability
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Reason *
C. Subject withdrawn
Whole body medical examinations of the subjects were conducted by a medically qualified
person on duty after subject admission, prior to dosing of study drug, once in every 12 hours post
dose in in-house part of the study and at discharge. No major complaints were observed in any
subjects in both the periods of the study.
Blood glucose level was monitored at the interval of 1 hrs after drug administration till 4 hrs of
drug administrations.
Subject No. 1 2 3 4
Subject No. 1 2 3 4
The study subjects were asked to inform the clinical staff of occurrence of any AEs immediately
once experienced. Furthermore, the clinical staff was instructed to check on the subjects for the
occurrence of any AE at specified time intervals (before dosing and 4.00, 8.00, 12.00 and 24.00
hrs from study drugs administration) and to notify immediately to the Clinical Investigator.
Table below describes the incidence of adverse events observed from the time of admission of
drug till the time of discharge. No Adverse Event was reported in both the Periods of the Study.
Lactic Acidosis - -
Abdominal discomfort - -
Diarrhea - -
Gas - -
Headache, - -
Indigestion - -
Nausea - -
Vomiting - -
Weakness - -
Others - -
Both the formulations (Test and Reference) were well tolerated. No AE/SAE/Death was reported
during the conduct of study.
5.10 Discussion
Bioequivalence study is usually carried out, by comparing the in vivo rate and extent of drug
absorption, of test and reference formulation, in healthy subjects.
The present study was an open label, balanced, randomized, two treatments, two periods, two
sequence, single dose, cross over bioequivalence study under fasting condition.
The following steps were incorporated into the study to minimize the bias:
In this particular study, study participants received test and reference products on separate
occasions, as a single dose, with random assignment to the two possible sequences of product
administration. Samples of plasma were analyzed for drug concentrations, and pharmacokinetic
parameters were obtained from the resulting concentrationtime curves. These pharmacokinetic
parameters were then analyzed statistically to determine whether the test and reference products
yielded comparable values.
The statistical methodology was based on the calculation of a 90% confidence interval for the
ratio (or difference) between the test and reference product pharmacokinetic variable averages.
The limits of the observed confidence intervals were within a predetermined range for the ratio
(or difference) of the product averages.
An Analysis of variance (ANOVA) test was performed (=0.05) on the untransformed and ln-
transformed pharmacokinetic parameters Cmax, AUC0-t and AUC0-. Factors included for
Analysis of variance (ANOVA) model were randomization sequence, subjects included in that
particular sequence, period and treatment effect. These factors for both untransformed and ln-
transformed pharmacokinetic parameters were found to be statistically insignificant.
Since, this was a DCGI submission study, so all the target parameters were chosen in accordance
with Guidelines for Bioavailability and Bioequivalence Studies issued by Central Drugs
Standard Control Organisation (CDSCO), March 2005.
The study was conducted as per the IEC approved study protocol, CDSCO BA/BE Guidelines,
March 2005, ICH-GCP Guidelines, Schedule Y to the Drugs and Cosmetic Rules, Indian GCP
Guidelines issued by CDSCO, GLP, Ethical Guidelines for Biomedical research on human
subjects issued by Indian Council of Medical Research (ICMR Guidelines) and Declaration of
Helsinki.
Hippocrates Independent Ethics Committee (HIEC), New Delhi has given the approval to
conduct this study and allowed to enroll twelve subjects for this particular study. HIEC were
regularly informed regarding the updates of this study.
Food and Drug Administration (FDA) provided the draft guidance on Metformin Hydrochloride
SR 500 mg for conducting the bioequivalence study (FDA Draft Guidance on Metformin, 2008).
It described that bioequivalence study of Metformin should be conducted in fasting condition
and design should be single-dose, two-way, crossover in-vivo in normal healthy males and
females, general population. On the basis of these considerations, a single-dose, two-treatment,
two-period, two- sequence crossover bioequivalence study on healthy human male normal
subjects was adopted.
As a result to its crossover design, no separate control group was necessary. 500 mg Metformin
Hydrochloride (SR) dose was expected to be reasonably well tolerated. It was also expected to
provide adequate plasma concentrations to be measured for assay.
The choice of the blood sampling time schedule was intended to cover the expected time of the
peak plasma concentrations of metformin, which was at around 7(4-8) hour (Drugs @ FDA,
2008). It was also extended for sufficient time to be able to obtain a reliable estimate of the
extent of absorption. The last sample collection was at 24.00 hours post-dose, which was more
than 3 times the maximum reported half-life of metformin 6.2 hours (Drugs @ FDA, 2008), and
covered at least 80% of the AUC0-.
Pharmacokinetic parameters like Cmax, tmax, t. AUC0-t, AUC0-, & Kel were calculated using
plasma concentration vs. time profile (actual time of sample collection) data of both
investigational products in individual subjects using WinNonlin Version 5.2. These data were
statistically analyzed and described.
Cmax is the measure that represents the major drug concentration observed and is directly
proportionate to the total drug amount absorbed by the body. The Cmax is generally higher when
the extent of drug bioavailability is greater. When comparing drug products, t max can be used as
an approximate indication of drug absorption rate. The value for tmax will become smaller
(indicating less time required to reach peak plasma concentration) as the absorption rate for the
drug becomes more rapid (Shargel & Andrew, 2004).
When pharmacokinetics parameters of the test drug of Sponsor were compared with reference
drug Dibeta SR 500 mg tablet formulation, it was found that Sponsor generic version have
almost same pharmacokinetic parameters.
Administration of the sponsors generic tablet produced the maximum plasma concentration of
621.18 112.09 ng/ml (Cmax) at the time 5.25 1.65 hour (tmax) whereas Dibeta SR tablet
(reference drug) peak concentrations were typically 621.10 126.65 ng/ml at the time 4.83 1.46
hour (tmax) following a single and a repeated 500 mg once daily dose, respectively. In adults,
peak plasma concentrations were achieved 7 (4-8) hour after administration of the oral tablet
(Drugs @ FDA, 2008).
The geometric least square mean ratios and the 90% CI of Cmax, AUC0-t and AUC0- for the test
product and the reference standard of the study drug were considered, for evaluating
bioequivalence. To be considered bioequivalent, ratios and confidence interval should lie within
the acceptance range of 80-125%.
Since, Pharmacokinetic Bioequivalence Parameters (Cmax, AUC0-t, AUC0-) of the Test Product
and the Reference Standard of the Study Drug were within the acceptance range of 80-125%. So
this study proves that, the test product Metformin hydrochloride (SR) 500 mg tablet is
Department of Pharmacy Practice 118
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The present study was an open label, balanced, randomized, two treatment, two period, two
sequence, single dose, cross over bioequivalence study under fasting condition; conducted at
Auriga Research Ltd., New Delhi. Metformin Hydrochloride (SR) 500 mg tablet was the Test
formulation and Dibeta SR tablet [containing Metformin Hydrochloride (SR) 500 mg] was used
as the Reference standard.
As per CDSCO BA/BE Guidelines, the minimum number of subjects for bioequivalence study
should not be less than sixteen unless justified for ethical reasons. Hippocrates Independent
Ethics Committee (HIEC), New Delhi has given the approval to Auriga Research Ltd. to conduct
this study and allowed to enroll twelve subjects for this particular study based on ethical reason.
The 90% confidence interval for ln-transformed pharmacokinetic parameters (Cmax, AUC0-t and
AUC0-) of the Test and Reference formulation of the Study drug Metformin Hydrochloride (SR)
500 mg were 89.13%-112.44%, 87.46%-123.85%, 88.29%-123.87% respectively. The Mean
Test/Reference ratio (%) for ln-transformed Cmax, AUC0-t and AUC0- were 100.110%, 104.080%
and 104.580% respectively.
Results showed that ln-transformed pharmacokinetic parameters (Cmax, AUC0-t and AUC0-) were
within the acceptable range of 80125%. So, the present study concludes that the Test
formulation Metformin Hydrochloride (SR) 500 mg tablet of Sponsor is bioequivalent to
Reference standard Dibeta SR tablet [containing Metformin Hydrochloride (SR) 500 mg] of
Torrent Pharmaceuticals Ltd., India.
7. References
1) Guidance for Industry: Bioavailability and Bioequivalence Studies for Orally Administered
Drug Products-General Considerations. U.S. Department of Health and Human Services,
Food and Drug Administration (FDA), Centre for Drug Evaluation and
11) U.S. Department of Health and Human Services Food and Drug Administration Center for
Drug Evaluation and Research (CDER). Guidance for Industry Bioavailability and
Bioequivalence Studies for Orally Administered Drug Products General Considerations.
Available at
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidan
ces/ucm070124.pdf [Cited on 2009 Nov 18]
12) Manual for Good Bioavailability and Bioequivalence Practices, Volume I Module 3:
Statistical Phase. 1st ed. Brazil; 2002. p. 13-19.
13) BCC Research. World Pharmaceutical Market, Report Code: PHM037A, Published: April
2009.Available at http://www.bccresearch.com/report/PHM037A.html. [Cited on 2010 Apr
3]
14) Central Drugs Standard Control Organization, Directorate General of Health Service,
Ministry of Health & Family Welfare, Government of India. Guideline for Bioavailability
& Bioavailability Studies, March 2005. Available at http://www.cdsco.nic.in/ [cited 2009
Aug 19]
15) Clinical Research Organization-India. Available at http://www.neeman-
medical.com/resources/clinicalresearch/dmc/edc.pdf [Cited 2010 April 19]
16) DiPiro JT, Talbert RL, Yee GC, Matzke GR. Pharmacotherapy: A Pathophysiologic
Approach, 6th ed. New York: MCGRAW-HILL Medical Publishing Division; 2005.
Ch.72, Diabetes Mellitus; p. 1335-36.
17) World Health Organization. Definition, diagnosis and classification of diabetes Mellitus
and its complications-Report of a WHO Conclusion. Geneva: World Health Organization;
1999.
18) DiPiro JT, Talbert RL, Yee GC, Matzke GR. Pharmacotherapy: A Pathophysiologic
Approach, 6th ed. New York: MCGRAW-HILL Medical Publishing Division; 2005.
Ch.72, Diabetes Mellitus; p. 1337-42.
19) Tripathi KD. Essentials of Medical Pharmacology, 6th ed. New Delhi: Jaypee Brothers;
2008. Ch. 19, Insulin, Oral Hypoglycaemic Drugs And Glucagon; p. 266-70.
20) DiPiro JT, Talbert RL, Yee GC, Matzke GR. Pharmacotherapy: A Pathophysiologic
Approach, 6th ed. New York: MCGRAW-HILL Medical Publishing Division; 2005.
Ch.72, Diabetes Mellitus; p. 1343-56.
21) FDA label of reference listed drug of Metformin hydrochloride SR. Available at
http://www.accessdata.fda.gov/scripts/cder/drugsatfda/. [Cited on 2009 Nov 15]
22) Michael RM, Dean TE, Sumit RM, James DL, Bhagra S, Pardeep S J, Mark CP, John J V,
John RP, Finlay AM. Diabetes care. 2010 Jun; 33(6): 1213-8
23) Donnelly LA, Morris AD, Pearson ER. Diabetes, obesity & metabolism. 2009 Apr; 11(4):
338-42.
24) Shelley RS, Nicholas SB, Justin AK, Edwin ES. The American journal of medicine. 2008
Feb;121(2): 149-157.e2
25) Information available on Toxnet drug information site. Available at
http://www.toxnet.nlm.nih.gov/ [Cited on 2009 Nov 17]
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 12.0 18.0 24.0
Screening
Incl./Excl.
Critaria
2 3 4 5 6 7 8 9 10 11 12 13 14
Day -21 to 0 Predose
The following is a representative time schedule for one subject assuming that the study
medication was administered at 09:00. There was a gap of 07 days between two
periods. Timings for other subjects were uniformly staggered:
Review of
X X - -
inclusion/exclusion criteria
Demographic examination X - - -
Clinical examination X X X -
Urine analysis X - - -
Electrocardiogram X - - -
Chest X-Rays* X - - -
Concomitant medications X X X -
*Chest X-Ray will be taken for those volunteer whose X-Ray was taken more than 6
months prior to the dosing of Period 01.
**X refers that the corresponding activity has been done during the assessment.
146-150 43 54 43 56 44 59 45 60 45 60 45 61
151-155 45 56 44 58 45 62 47 63 47 64 47 64
156-160 48 59 48 61 48 65 50 67 50 67 50 68
161-165 51 62 51 65 51 69 53 70 53 71 53 72
166-170 54 66 54 69 55 73 56 75 56 75 56 76
171-175 58 71 58 73 58 78 60 80 60 80 60 81
176-180 61 75 62 78 62 83 64 85 64 86 64 87
181-185 65 80 65 83 66 88 63 90 68 92 68 93
186-190 69 85 69 89 70 94 72 96 72 97 72 98
Height (cm) 145 150 155 160 165 170 175 180 185
Study Title: An open label, balanced, randomized, two treatment, two period, two sequence,
cross over, bioequivalence study of a single dose of Metformin Hydrochloride (SR) 500 mg
tablet of Sponsor Cipla Ltd and Dibeta SR (containing Metformin Hydrochloride (SR) 500 mg)
tablet of Torrent Pharmaceuticals Ltd, India, in healthy human adult male subjects under fasting
condition.
1. I confirm that I have read and understood the information provided to me regarding the study [ ]
and have had the opportunity to ask questions.
3. The general purpose of the experiment has been explained to me and I am convinced that it is [ ]
for the benefit of science and mankind.
Ingestion of Drugs
9. I have informed my family regarding my participation in this trial and I understand that I can [ ]
withdraw from the study at any time. I have been informed that i will be compensated for my
participation
Name of the Person Obtaining ICF Signature / Date of the Person obtaining
ICF
ADVERSE EVENT
Adverse Event Start Date Reporting Date and Time Method of Reporting:
and Time Spontaneous
Date: Date: Observed
Elicited
Time: Time:
Description of Adverse Event Including Medication and Advice
This section is to be filled in the last either by Clinical Investigator or Principal Investigator
Please choose and mark one of each below to rate the event
Adverse Event End Date and Time: No. of follow-up sheets used:
Seriousness Frequency Severity Relationship to Action taken Outcome
1 = Serious 1 = isolated 1 = mild Study drug 1 = none 1 = recovered
2 = Non 2 = intermittent 2 = moderate 1 = certain 2 = medication without sequelae
Serious 3 = continuous 3 = severe 2 = probable / therapy 2 = recovered
4 = unknown 4= Life- likely 3 =procedure with Sequelae*
Expectedne threatening 3 = possible 4 = hospitalization 3 = ongoing
ss: 4 = unlikely 5 = other* 4 = death
1= Expected 5 = unclassifiable 5 = other*
2= 6= unclassified
Unexpected
A. Subject Information
Enrollment No.: Gender: Male Female Date of Birth:
Age: (completed years) Height: (cm) Weight: (kg)
B. Study Drug (s)
Drug (generic/proprietary Route Duration of Therapy Indication of
name) and Dosage Form / From To Treatment
Dose(e.g. Tab 10 mg OD)
C. Adverse Reaction
Adverse Event Start Date / Time: Adverse Event End Date / Time:
OR Ongoing
Duration of Adverse Event: (days/hours/minutes)
Adverse Event Summary: Diagnosis (if known)
Include clinical history of event, associated signs & symptoms, alternative etiologies being
considered, medical management and relevant past medical history below or attach
additional pages if necessary.
(Principal Investigator)
Continued.
Abstract
Intensity
Mild Moderate Severe Life- threatening
Please indicate SAE category from the following choices:
Death Persistent or significant disability or incapacity Congenital anomaly or birth defect
life threatening Inpatient hospitalization or prolongation of existing hospitalization
Important medical event (Any event that, based upon appropriate medical judgment, may jeopardize the patient and
may require medical or surgical intervention to prevent one of the outcomes listed above).
Comments:
(Principal Investigator)
Abstract