Bioequivalence protocol 46

1. BIOEQUIVALENCE PROTOCOL DELIBRATED BY: UNDER THE GUIDANCE OF J.RAJESH D r.SATYABRATA BHANJA M.Pharm,Ph.D M.PHARM(CEUTICS) DEPARTMENT OF PHARMACEUTICS 256213886012 MALLARAEDDY COLLEGE OF PHARMACY

2. Bioequivalence : It is a relative term which denotes that the drug substance in 2 or more identical dosage form,reaches the systemic circulation at the same relative rate and to the same relative extent i.e. Their plasma concentration –time profiles will be identical without significant statistical differences .

3. Elements of Bioequivalence Study Protocol 1.Title a.Principal investigator b.Project number and date 2.Study objective 3.Study design a.Design b.Drug Products i. Test products(s) ii.Reference product c.Dosage regimen d.Sample collection schedule

4. e.Housing f.Fasting /Meals schedule g.Analytical methods 4.Study population a.Subjects b.Subject selection i.Medical history ii.Physical examination iii.Laboratory tests c.Inclusion/exclusion criteria i.Inclusion criteria ii.Exclusion criteria

5. d.Restrictions/prohibitions 5.Clinical procedures a.Dosage and Drug Administration b.Biological sampling schedule c.Activity of subjects 6.Ethical considerations a.Basic principles b.Institutional review board c.Informed consent d.Indication for subject withdrawl e.Adverse reaction and emergency procedures

6. 7.Facilities 8.Data Analysis a.Analytical validation procedure b.Statistical treatment of data 9.Drug Accountability 10.Appendix

7. Study Objective: The objective for a bioequivalence study is that the drug bio availability from test and reference products are not statiscally different when administered to patients are subjects at same molar dose under similar experimental conditions.

8. Objectives for Bioequivalence Studies : Equivalence : It is a relative term that compares drug products with respect to a specific characteristic or function to a defined set of standards. There are several types of equivalences Pharmaceutical Equivalence : This term implies two or more drug products are identical in strength,quality purity ,content uniformity,and disintegration and dissolution characteristics. Chemical equivalence: It indicats that two or more drug products contained the same labelled chemical substance as an active ingrediant in the same amount.

9. Therapeutic equivalence : This term indicats that two or more drug products that contain the same therapeutically active ingrediant elicit identical pharmacological effects and can control the disease to the same extent. Types of Bioequivalence studies : Bioequivalence can be demonstrated either In vivo,or In vitro In vivo Bioequivalence Studies: 1.Oral immediate – release products with systemic action 2.Non oral immediate – release produts. 3.Modified- release products with systemic action

10. In vitro Bioequivalence studies : 1.The drug products differs only in strength of the active substances. 2.The drug products meets of the following requirements. The product is in the form of solution (elixir,syrup,tincture). The product contains active ingrediant in the same concentrasation approved drug product .

11. Study Designs: For many drug products,the FDA, division of bio equivalence, office of generic drugs provides guidances for the performances of in-vitro dissolution and in-vivo bioequivalence studies Generally two bioequivalence studies required for solid oral dosage forms including

12. 1.A fasting study 2.A food intervention study 1.Cross Over Study Design : - Two formulations, even number of subjects - Randomly divided into two equal groups

13. First Period: Each number of one group receive a single dose of the test formulation and each member of the other group receive the standard formulations. Subject Period 1 Period 2 1-8 Test Standard 9-16 Standard Test

14. 2.Latin Square Design : - More than two formulations Eg: A group of volunteers will receive formulations in the sequence . Volunteer Number Period 1 Period 2 Period 3 1 A B C 2 B C A 3 C A B

15. 3.Balance Incomplete Block Design: - More than 3 formulations - Latin square design will not be used because each volunteer may required drawing of too many many blood samples If each volunteer expected to receive atleast 2 formulations then such study can be carried out using BIBD.

16. Volunteer Number Period 1 Period 2 1 A B 2 A C 3 B D 4 B C 5 C D 6 B D 7 C A 8 D A 9 C B 10 D B 11 D C 12 A B

17. 4.Parallel Group Design : - Even number of subjects in 2 groups -Each receive a different formulation -No wash out necessary -For drugs with long half life Treatment A Treatment B 1 2 3 4 5 6 7 8 9 10 11 12

18. 5.Replicate Cross Over Study Design : For highly variable drugs Allows comparisions of with in subject variances Reduces the number of subjects needed 4-periods, 2-sequences, 2 formulations design(recommended) 3-sequences, 3-periods, single dose partially replicated

19. Peeriod 1 Period 2 Period 3 A B C B C A C A B A C B B A C C B A Period 1 2 3 4 Group 1 Test Reference Test Reference Group 2 Reference Test Reference Test

20. Difference Between Parallel Group And Cross Over Study Design: Parallel Group Cross Over Groups assigned different treatments Each patient receives both treatments Shorter duration Longer duration Larger sample size Smaller sample size No carry over effect Carry over effect

21. 6.Pilot Study: If the sponsor chooses in a small number of subjects To access a variability, optimise the sampla collection time intervals, and provide other information Eg: Immediate release products – careful timing of intial samples-avoid a sub sequent finding that the first samole collection, occured after the plasma concentration peak. Modified released products: To determine sampling shedule- Assess log time and dose dumping

22. Analytical Methods: Analytical methods used in an in-vivo bio availability, bio equivalence, or pharmacodynamic studies must be validated for accuracy and sufficient sensitivity. The analytical method for measurement of drug must be validated for accuracy, precision, sensitivity, specificity, and robustness. The use of more than one analytical method during a bio equivalence study may not be valid because different methods may yield different values.

23. Subject Selection: Healthy adult volunteers age 18-45years age/sex representation corresponding to therapeutic and safety profile weight with in normal limits women-pregnency test period to first and last dose of study Selection Of Number Of Subjects: Sample size estimated by Pilot Experiments Previous Studies Published Data

24. Significance level desired usually 0.05 Power of study normally 80% or more Minimum 16 subjects unless ethical justification Allow for drop outs Exclusion Criteria : H/o allergy to test drug H/o liver or kidney disfunction H/o jaundice in past 6 months Chronic diseases Eg: asthma, arrthiritis Psychiatric illness

25. Administration of drug products: Administration of drug products to the should be based on randamization.After the administration of drug products, biood samples are withdrawn from the subjects at fixed time points. It takes some to take a sample from each subject, and the total time difference between first and last subject ay range from 10 to 20 minutes depending upon the number of subjects and technicians in the study. This 10 to 20 minutes difference would represent a substantial change in the drug concentrations observed in the blood.

26. If under these conditions treatments are administered to the the subjects in a seqential manner( such as teatment A to the first 6 volunteers,teatment B to volunteers 7 to 12, and teatment C to Volunteers 13 to18),the error between the time of administration and sampling will gradually increase from treatment group to treatment group. This is because of sequential administration of drug products to different treatments.

27. Sampling: The biological sample to be used in the study as to be decided before the commencement of a bioavailability study. If the bioavailability of a given dosage form is to be evaluated by a blood level study, some estimate of the area under the serum concentration v/s time curve, peak serum concentration,time of peak concentration must be obtained from the study. These factors can markedly influence the ‘apparent’ results obtained in a given study.

28. The sampling scheme should frequent enough to define the absorption phase, the peak, and the elimination phase during a drugs time course in the body. The absorption rate, volume of distribution, elimination rate, all influence the apparent drug concentration one obtains in a given sample. It is necessary to see that all these factors influence each dosage form equally. To estimate the AUC from the data, sampling as to be carried out till the concentration of the drug reaches the linear elimination phase.

29. For first-order process , the time necessary for a complete elimination would be infinity. A rule of thumb sampling in a blood level study is to sample for 3-5 half lifes of the drugs. If half life is not known , sampling should proceed untill 1/10 or 1/20 of the peak levels are reached. In the case of urinary excretion studies, the same principles apply. The advantage of urinary excretion studies are 1.it involes non-invasive method of sampling. 2.the drug concentration in the urine is greaterthan blood/serum allowing easy estimation of the drug.

30. The amount of drug excreted in urine is obtained directly. In the case of a blood level study, the amount of drug in the body is estimated using pharmacokinetic parameters. The urinary excretion method has several disadvantages 1.urinary excretion studies are not useful in estimating the drug absorption rate. 2.In some cases, the metabolites of the drug are also concentrated in the sample that interferes with the estimation of unchanged drug in the urine sample.

31. Evaluation of data: Pharmacokinetic evaluation of the data : F or single dose studies, including a fasting study or a food intervention study, the pharmacokinetic analyses include calculation for each subject of the area under the curve to the last quantifiable concentration (AUC ) and to infinity (AUC ), t max and C max . Additionally ,the elimination rate constant,k, the elimination half-life,t 1/2 ,

32. Statistical evaluation of the data: Bioequivalance is generally determined using a comparision of population averages of a bioequivalance metric,such as AUC and C max . This approach, termed average bioequivalence,involves for the ratio of averages of the test and reference drug products.

33. Statistical Analysis For Average Bio equivalence: Based on log transformed data Point estimates of the mean ratios Test / reference for AUC and C max are between 80% -125% AUC and C max 90% confident intervals must fit between 80%-125% Statistical model typically includes factors accounting for following sources of variations: Sequence, subjects, nested in sequences, period in treatment

34. Proposed And Contents Of An In vivo Bio equivalence Study Submission And Accompaning In vitro Data: Title Page Study Title Name of sponsor Name and Address of clinical laboratory Name of Principal Investigator(S) Name of Clinical Investigator Name of Analytical Laboratory Dates of Clinical Study Signature of principal investigator(and date) Signature of Clinical Investigator(and date)

35. Table Of Contents 1.Study Resume Product Information Summary of Bio equivalence study Summary of Bio equivalence data Plasma Urinary Excreation Figure of mean plasma concentration-time profile Figure of mean cumulative urinary excreation Figure of mean urinary excreation rates 2.Protocol And Approvals Protocol Letter of acceptance of protocol from fda Informed consent form Letter of approval of institutional review board 3.Clinical study Summary of Study Details of study Demographic characteristics of the subjects Subject assignement in the study

36. Mean physical characteristics of subjects arranged by sequence Details of clinical activity Deviation from protocol Vital science of subjects Adverse reactions report 4.Assay Methodology And Validation Assay method discription Validation procedure Summary of validation Data on linearity of standard samples Data on interday precision and accuracy Data on intraday precision and accauracy Figure for standard curve for low/high ranges Chromatograms of standard and quality control samples Sample calculation

37. 5.Pharmacokinetic Parameters and Tests Definitions and calculaton Statistical tests Drug levels at each sampling time and pharmacokinetic parameters Figure of mean plasma concentration-time profile Figure of individual subjects plasma concentrations-time profiles Figure of mean cumulative urinary excreation Figures of individual subject urinary excreation rates Tables of individual subject data arranged by drug, drug/period, drug/sequence

38. 6.statistical analyses statistical considerations summary of statistical significance summary of statistical parameters analysis of variance,least squares estimates and least squares means assessment of sequence, period, and treatment effects 90% confidence intervals for the differences between test and reference products for the log-normal-transformed parameters of AUC 0-t , AUC 0-infinty , C MAX should be 80%-125%.

39. 7. appendices Randamization schedulule sample identification codes analytical raw data chromatograms of at least 20% of subjects medical records and clinical reports clinical facilities discription analytical facilities discription curricula vitae of investigators 8. invitro testing dissolution testing dissolution assay methadology content uniformity testing potency determination

40. 9. batch size and formulations batch record quantitative formulations

41. Applications of pharmacokinetics in drug development,NDDS: Drug research: New Drug i.Discovery ii.Development Drug Discovery :- Hypothesis of target enzyme on receptor for particular disease Suitable models Screening of new drug Screening of in vitro/in vivo biological activity Drug development : Empirical Dosage regimen Semi empirical Dosage regimen Structaral activity These are development of chemical structures.

42. Emprical Dosage Regimen :- It is designated by the physician based on emprical clinical data,personal experience and clinical observations.This approach is however,not very accurate Semi emprical Dosage regimen : It is most accurate approach and is based on the pharmacokinetics of drug in the individual patient.The approch is suitable for hospitalised patients but is quite expensive. Pharmacokinetics-AUC,C MAX ,T MAX . Pharmacodynamics-On set of action, On set of time,MSC,MEC,MTC.

43. Applications: Design of new drug development: Design a level of optimum formalation for better use of drug Design a level of control and sustained released formulation. Selection of appropriate new drug administration then the selection of right drug for the particular disease Design and development of drug design,processing dosage regimen Study of in vitro, in vivo studies Study of bioequivalant studies. The study of pharmacokinetics and pharmacodynamic relationship Development of rational drug design,development rational dose frequency and duration.

44. Determaine the drug drug interactions Design of appropriate multiple dosage regimen Therapeutic dose of indiviual drug Pharmacokinetics charactarization of drug selection of the suitable novel drug delivary system. NDDS parameters are the i.T 1/2 ii.T 90 iii.Elimination rate constant iv. Area under curve v.Volume of distibution vi.Steady state concentration vii.Mean residance time ix.Dosage form index x.Relative area xi.Absorption rate

45. References: BIOPHARMACEUTICS AND PHARMACOKINETICS second edition V.Venkateswarlu APPLIED BIOPHARMACEUTICS & PHARMACOKINETICS sixth edition Leon Shargel Susanna wu-Pong Andrew yu

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