FINAL PROJECT LEVOFLOXACIN

NEW DRUG DEVELOPMENTAL & APPROVAL PROCESS

LEVOMAX

(Levofloxacin)

Submitted By: Zohaib Ahmad(Roll#14)

Submitted to:

Jalwaz Tihami(Roll#20)

Sir Ikram Ullah Khan

Azeem Imam(Roll#25)

B-Pharm, M.Phil, MS (TQM), R.Ph.

Rizwan Rashid(Roll#43) Ali Tariq(Roll#136)

COLLEGE OF PHARMACY GC UNIVERSITY FAISALABAD

CONTENTS CHAPTER.NO.01: INTRODUCTION CHAPTER.NO.02: FLOW CHART OF NEW DRUG DEVELOPMENT CHAPTER.NO.03: SOURCE AND SYNTHESIS CHAPTER.NO.04: PRECLINICAL STUDIES CHAPTER.NO.05: PREFORMULATION CHAPTER.NO.06: INVESTIGATION NEW DRUG APPLICATION CHAPTER.NO.07: CLINICAL TRIALS CHAPTER.NO.08: LONG TERM ANIMAL TOXICITY STUDIES CHAPTER.NO.09: PRODUCT FORMULATION CHAPTER.NO.10: MANUFACTURING AND CONTROL CHAPTER.NO.11: PACKAGE AND LABEL DESIGN CHAPTER.NO.12: NEW DRUG APPLICATION CHAPTER.NO.13: POST MARKETING SURVIELLANCE CHAPTER.NO.14: PRODUCT LINE EXTENTION

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CHAPTER#01 INTRODUCTION

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INTRODUCTION OF DRUG DISCOVERY AND DEVELOPMENT INTRODUCTION: Discovering and bringing one new drug to the public typically costs a pharmaceutical or biotechnology company nearly $900 million and takes an average of 10 to 12 years. In special circumstances, such as the search for effective drugs to treat AIDS, the U.S. Food and Drug Administration (FDA) has encouraged an abbreviated process for drug testing and approval called fast-tracking. The drug discovery and drug development process is designed to ensure that only those pharmaceutical products that are both safe and effective are brought to market. PPD provides a broad array of drug discovery and development services and products to pharmaceutical, biotechnology and medical device companies to expedite drug development, from drug discovery through clinical studies and post-approval support. Drug development is a blanket term used to define the entire process of bringing a new drug or device to the Market. It includes Drug discovery / product development, pre-clinical research (microorganisms/animals) and Clinical trials (on humans). Few people still refer to the drug development as mere preclinical development. HOW ARE NEW DRUGS DISCOVERED? New drugs begin in the laboratory with scientists, including chemists and pharmacologists, who identify cellular and genetic factors that play a role in specific diseases. They search for chemical and biological substances that target these biological markers and are likely to have drug-like effects. Out of every 5,000 new compounds identified during the discovery process, only five are considered safe for testing in human volunteers after preclinical evaluations. After three to six years of further clinical testing in patients, only one of these compounds is ultimately approved as a marketed drug for treatment. The following sequence of research activities begins the process that results in development of new medicines: •

Target Identification. Drugs usually act on either cellular or genetic chemicals in the body, known as targets, which are believed to be associated with disease. Scientists use a variety of techniques to identify and isolate 4

individual targets to learn more about their functions and how they influence disease. Compounds are then identified that have various interactions with the drug targets that might be helpful in treatment of a specific disease. •

Target Prioritization/Validation. To select targets most likely to be useful in the development of new treatments for disease, researchers analyze and compare each drug target to others based on their association with a specific disease and their ability to regulate biological and chemical compounds in the body. Tests are conducted to confirm that interactions with the drug target are associated with a desired change in the behavior of diseased cells. Research scientists can then identify compounds that have an effect on the target selected.

•

Lead Identification. A lead compound or substance is one that is believed to have potential to treat disease. Laboratory scientists can compare known substances with new compounds to determine their likelihood of success. Leads are sometimes developed as collections, or libraries, of individual molecules that possess properties needed in a new drug. Testing is then done on each of these molecules to confirm its effect on the drug target.

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Lead Optimization. Lead optimization compares the properties of various lead compounds and provides information to help biopharmaceutical companies select the compound or compounds with the greatest potential to be developed into safe and effective medicines. Often during this same stage of development, lead prioritization studies are conducted in living organisms (in vivo) and in cells in the test tube (in vitro) to compare various lead compounds and how they are metabolized and affect the body.

WHAT IS REQUIRED BEFORE AN INVESTIGATIONAL DRUG CAN BE TESTED IN HUMAN VOLUNTEERS? In the preclinical stage of drug development, an investigational drug must be tested extensively in the laboratory to ensure it will be safe to administer to humans. Testing at this stage can take from one to five years and must provide information about the pharmaceutical composition of the drug, its safety, how the drug will be formulated and manufactured, and how it will be administered to the first human subjects.

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•

Preclinical Technology. During the preclinical development of a drug, laboratory tests document the effect of the investigational drug in living organisms (in vivo) and in cells in the test tube (in vitro).

•

Chemistry Manufacturing and Controls (CMC)/Pharmaceutics. The results of preclinical testing are used by experts in pharmaceutical methods to determine how to best formulate the drug for its intended clinical use. For example, a drug that is intended to act on the sinuses may be formulated as a time-release capsule or as a nasal spray. Regulatory agencies require testing that documents the characteristics -- chemical composition, purity, quality and potency -- of the drug's active ingredient and of the formulated drug.

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Pharmacology/Toxicology. Pharmacological testing determines effects of the candidate drug on the body. Toxicology studies are conducted to identify potential risks to humans.

Results of all testing must be provided to the FDA in the United States and/or other appropriate regulatory agencies in order to obtain permission to begin clinical testing in humans. Regulatory agencies review the specific tests and documentation that are required to proceed to the next stage of development. HOW ARE INVESTIGATIONAL DRUGS TESTED IN HUMANS? Testing of an investigational new drug begins with submission of information about the drug and application for permission to begin administration to healthy volunteers or patients. Investigational New Drug (IND)/Clinical Trial Exception (CTX)/Clinical Trial Authorization (CTA) Applications. INDs (in the U.S.), CTXs (in the U.K.) and CTAs (in Australia) are examples of requests submitted to appropriate regulatory authorities for permission to conduct investigational research. This research can include testing of a new dosage form or new use of a drug already approved to be marketed. In addition to obtaining permission from appropriate regulatory authorities, an institutional or independent review board (IRB) or ethical advisory board must approve the protocol for testing as well as the informed consent documents that

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volunteers sign prior to participating in a clinical study. An IRB is an independent committee of physicians, community advocates and others that ensures a clinical trial is ethical and the rights of study participants are protected. Clinical testing is usually described as consisting of Phase I, Phase II and Phase III clinical studies. In each successive phase, increasing numbers of patients are tested. •

Phase I Clinical Studies. Phase I studies are designed to verify safety and tolerability of the candidate drug in humans and typically take six to nine months. These are the first studies conducted in humans. A small number of subjects, usually from 20 to 100 healthy volunteers, take the investigational drug for short periods of time. Testing includes observation and careful documentation of how the drug acts in the body -- how it is absorbed, distributed, metabolized and excreted.

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Phase II Clinical Studies. Phase II studies are designed to determine effectiveness and further study the safety of the candidate drug in humans. Depending upon the type of investigational drug and the condition it treats, this phase of development generally takes from six months up to three years. Testing is conducted with up to several hundred patients suffering from the condition the investigational drug is designed to treat. This testing determines safety and effectiveness of the drug in treating the condition and establishes the minimum and maximum effective dose. Most Phase II clinical trials are randomized, or randomly divided into groups, one of which receives the investigational drug, one of which gets a placebo containing no medication and sometimes a third group that receives a current standard treatment to which the new investigational drug will be compared. In addition, most Phase II studies are double-blinded, meaning that neither patients nor researchers evaluating the compound know who is receiving the investigational drug or placebo.

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Phase III Clinical Studies. Phase III studies provide expanded testing of effectiveness and safety of an investigational drug, usually in randomized and blinded clinical trials. Depending upon the type of drug candidate and the condition it treats, this phase usually requires one to four years of testing. In Phase III, safety and efficacy testing is conducted with several hundred to

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thousands

of

volunteer

patients

suffering

from

the

condition

the

investigational drug treats.

New Drug Application (NDA)/Marketing Authorization Application (MAA): NDAs (in the U.S.) and MAAs (in the U.K.) are examples of applications to market a new drug. Such applications document safety and efficacy of the investigational drug and contain all the information collected during the drug development process. At the conclusion of successful preclinical and clinical testing, this series of documents is submitted to the FDA in the U.S. or to the applicable regulatory authorities in other countries. The application must present substantial evidence that the drug will have the effect it is represented to have when people use it or under the conditions for which it is prescribed, recommended or suggested in the labeling. Obtaining approval to market a new drug frequently takes between six months and two years.

DOES TESTING CONTINUE AFTER A NEW DRUG IS APPROVED? After the FDA (or other regulatory agency for drugs marketed outside the U.S.) approves a new drug, pharmaceutical companies may conduct additional studies, including Phase IIIb and Phase IV studies. Late-stage drug development studies of approved, marketed drugs may continue for several months to several years. •

Phase IIIb/IV Studies. Phase IIIb trials, which often begin before approval, may supplement or complete earlier trials by providing additional safety data or they may test the approved drug for additional conditions for which it may prove useful. Phase IV studies expand testing of a proven drug to broader patient populations and compare the long-term effectiveness and/or cost of the drug to other marketed drugs available to treat the same condition.

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Post-Approval Studies. Post-approval studies test a marketed drug in new age groups or patient types. Some studies focus on previously unknown side effects or related risk factors. As with all stages of drug development testing, the purpose is to ensure the safety and effectiveness of marketed drugs 8

STEPS IN NEW DRUG DEVELOPMENT TILL NDA IS FILED

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Steps in New Drug Development till NDA is Filed

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CHAPTER#02 FLOW CHART FOR NEW DRUG DEVELOPMENT

FLOW

CHART

11

NEW CHEMICAL ENTITY

• •

•

Organic Synthesis Molecular Modification Isolation from Plants

PRECLINICAL STUDIES

• • •

•

Chemistry Physical Properties Biological Properties ADME Toxicology Preformulation

INVESTIGATIONAL NEW DRUG APPLICATION (IND)

•

•

Submission FDA Review

CLINICAL TRIALS

• •

•

PRECLINICAL STUDIES (Continued)

• • •

Phase І Phase І І Phase І І І

•

Long Term Animal Toxicity Product Formulation Manufacturing and Controls Package and Label Design

NEW DRUG APPLICATION (NDA)

• • •

•

Submission FDA Review Preapproval Plant Inspection FDA Action

POST MARKETING SURVEILLANCE

• • • •

•

Phase 4 clinical studies Additional Indications Adverse Drug Reporting Product Defect Reporting Product Line Extention

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CHAPTER#03 SOURCE AND SYNTHESIS

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SYNTHESIS OF LEVOFLOXACIN Levofloxacin is a synthetic compound and is synthesized as follow;

Procedure: 1,2-Cyclic sulfamidates undergo efficient and regiospecific nucleophilic cleavage with 2-bromophenols (and related anilines and thiophenols), followed by Pd(0)mediated amination to provide an entry to substituted and enantiomerically pure 1,4benzoxazines (and quinoxalines and 1,4-benzothiazines). This chemistry provides a short and efficient entry to (3S)-3-methyl-1,4-benzoxazine 19, a late stage intermediate in the synthesis of levofloxacin. This intermediate, through a series of steps, is than converted into levofloxacin.

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CHAPTER#04 PRECLINICAL STUDIES ™ Chemistry ™ Physical Properties ™ Biological Properties • Pharmacology • Pharmacokinetics • Toxicity

UNIT-1:

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CHEMISTRY OF LEVOFLOXACIN

IUPAC-Name: (S)-7-fluoro-6-(4-methylpiperazin-1-yl)-10-oxo-4-thia-1-azatricyclo [7.3.1.05,13] trideca-5(13),6,8,11-tetraene-11-carboxylic acid

Chemical Formula: (-) - (S)- 9 fluoro- 2, 3- dihydro -3- methyl- 10- (4- methyl- 1piperazinyl) –7 – oxo -7H – pyrido [1, 2, 3 -de]- 1, 4 benzoxazine- 6- carboxylic acid hemihydrate. Empirical Formula: C18H20FN3O4 ½H2O Routes: Oral, IV, ophthalmic Molecular Weight: 370.38.

Stable Coordination Compounds: Levofloxacin has the potential to form stable coordination compounds with many metal ions. This in vitro chelation potential has the following formation order: Al+3>Cu+2>Zn+2>Mg+2>Ca+2.

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Structural Formula of Levofloxacin

UNIT-2: 17

PHYSICAL PROPERTIES Appearance: Levofloxacin is a light yellowish-white to yellow-white crystal or crystalline powder. Stability: Stable under ordinary conditions Melting Point: 218 ºC (http://www.chemblink.com/products/100986-85-4.htm) Solubility: Insoluble in water The data demonstrate that from pH 0.6 to 5.8, the solubility of levofloxacin is essentially constant (approximately 100 mg/mL). Levofloxacin is considered soluble to freely soluble in this pH range, as defined by USP nomenclature. Above pH 5.8, the solubility increases rapidly to its maximum at pH 6.7 (272 mg/mL) and is considered freely soluble in this range. Above pH 6.7, the solubility decreases and reaches a minimum value (about 50 mg/mL) at a pH of approximately 6.9. Physical State: crystalline powder Odour: Odourless

UNIT 3:

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BIOLOGICAL PROPERTIES These biological properties are based on pre-clinical studies that are carried out in animals. (A) PHARMACOLOGY:

Pharmacotherapeutic Group: Quinolone Antibacterials, Fluoroquinolones Levofloxacin is a synthetic antibacterial agent of the fluoroquinolone class and is the S (-) enantiomer of the racemic drug substance ofloxacin.

Mechanism of action:(Chemical Basis) Levofloxacin is a broad-spectrum antibiotic that is active against both Gram-positive and Gram-negative bacteria. It functions by inhibiting DNA gyrase, a type II topoisomerase, and topoisomerase iv, which is an enzyme necessary to separate replicated DNA, thereby inhibiting cell division. The fluoroquinolones interfere with DNA replication by inhibiting an enzyme complex called DNA gyrase. In particular, some congeners of this drug family display high activity not only against bacterial topoisomerases but also against eukaryotic topoisomerases, and are toxic to cultured cells and in vivo tumor models. Although the quinolone is highly toxic to mammalian cells in culture, its mechanism of cytotoxic action is not known. Quinolone-induced DNA damage was first reported in 1986.

Pharmacokinetics: Absorption: 19

Bioavailability Approximately 99%. Rapidly absorbed from GI tract.Peak plasma concentrations usually attained 1-2 hours after an oral dose. Steady-state plasma concentrations attained within 48 hours with once-daily regimen.

Distribution: Extent Widely distributed into body tissues and fluids, including skin, blister fluid, and lungs. It is also distributed into CSF. Plasma Protein Binding 24-38% bound to serum proteins.

Elimination: Metabolism Undergoes limited metabolism to inactive metabolites. Elimination Route Eliminated principally as unchanged drug in urine. Approximately 87% of an oral dose eliminated in urine and lomefloxacin > rhodamine 123 > levofloxacin > ofloxacin). Based on comparison to

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labetalol, the high permeability internal standard, ciprofloxacin was classified as a low permeability drug, whereas lomefloxacin, levofloxacin, and ofloxacin were classified as high permeability drugs. The in vitro permeability results matched human in vivo data based on absolute bioavailabilities. This laboratory exercise demonstrated the applicability of an in vitro permeability method for classifying drugs as outlined in the BCS Guidance. Dissolution: Drug exhibit good dissolution properties. Method and Equipment: Rotating Basket Apparatus is used. Drug is placed in the Rotating Basket Apparatus and the dissolution of compound is determined

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UNIT-3: STABILITY ANALYSIS Hydrolysis: There is no effect of moisture. Stability-indicating RP-HPLC method for levofloxacin in the presence of degradation products, its process related impurities and identification of oxidative degradant The objective of current study was to develop a validated specific stability indicating reversed-phase liquid chromatographic method for the quantitative determination of levofloxacin as well as its related substances determination in bulk samples, pharmaceutical dosage forms in the presence of degradation products and its process related impurities. Forced degradation studies were performed on bulk sample of levofloxacin as per ICH prescribed stress conditions using acid, base, oxidative, water hydrolysis, thermal stress and photolytic degradation to show the stability indicating power of the method. RESULT: Significant degradation was observed during oxidative stress and the degradation product formed was identified by LCMS/MS, slight degradation in acidic stress and no degradation was observed in other stress conditions. The chromatographic method was optimized using the samples generated from forced degradation studies and the impurity spiked solution. Good resolution between the peaks corresponds to process related impurities and degradation products from the analyte were achieved on ACE C18 column using the mobile phase consists a mixture of 0.5% (v/v) triethyl amine in sodium dihydrogen orthophosphate dihydrate (25 mM; pH 6.0) and methanol using a simple linear gradient. The detection was carried out at 294 nm. The limit of detection and the limit of quantitation for the levofloxacin and its process related impurities were established. The stressed test solutions were assayed against the qualified working standard of levofloxacin and the mass balance in each case was in between 99.4 and 99.8% indicating that the developed LC method was stability indicating. Validation of the developed LC method was carried out as per ICH requirements. The developed LC method was found to be suitable to check the quality of bulk samples 33

of levofloxacin at the time of batch release and also during its stability studies (long term and accelerated stability).

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CHAPTER#06 INVESTIGATIONAL NEW DRUG APPLICTION

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Documents required by Ministry of Health for the approval of Clinical Trials in Pakistan

Documents Investigator Brochure. Final protocol. Informed Consent (English and Urdu )Form Fees 5000. (Head of Account) C-Non Tax Revenue C02- Receipts from Civil Administration and other Functions. C028-Social Services. C02841-Health-Other Receipts List of participating countries. Phase of Trial Quantity of drug to be imported on Form 4 of Drugs Import & Export Rules 1976 along with the sites where trial is to be conducted. CV’s of Investigators. Ethics committee approval of sites, with complete composition of committee i.e names and designation of members. GMP Certificate along with CPP/Free Sale Certificate of Country of Origin. Pre- clinical/ Clinical data/ Safety studies. Summary of the Protocol Summary of the IB ( for quick review on drug). Adverse Event Reporting Form . Number patients to be enrolled in each center Name of Monitors/ Clinical Research Associate Evidence of registration in country of origin Copy of registration letter if drug is registered in Pakistan Sample of label of drug Duration of Trial

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CHAPTER#07 CLINICAL TRIALS

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CLINICAL TRIALS

Our product is converted into a suitable dosage form. Than we submitted investigational to FDA. All preclinical studies mentioned and took permission for clinical trials.

PHASE-1 CLINICAL TRIALS: Aims: To study the safety of the drug in healthy volunteers No. of Patients: We selected 20-100 healthy voluntaries for phase 1 trials. Procedure: We administered 1/10 of no effect dose of animals. This purloins gets stable so we increased the dose gradually and checked the response. In this phase, Parmacokinetics and Pharmacodynamic studies of a drug are undertaken to determine its toxicity, metabolism, absorption, distribution and elimination and pharmacological action preferred route of administration and safe dosage. Duration: Phase-1 survives usually for 1-2 years. Results: We conducted study under careful circumstances by personals trained in clinical pharmacology. The clinical research on ranitidine is preceded to phase-2 because phase 1 studies showed promise and no drug reaction, which became evident.

PHASE-2 CLINICAL TRIALS: Phase II studies are sometimes divided into Phase IIA and Phase IIB. ¾ Phase IIA is specifically designed to assess dosing requirements (how much drug should be given).

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¾ Phase IIB is specifically designed to study efficacy (how well the drug works at the prescribed dose(s)). Aims: ¾ This trial aims to demonstrate conclusively efficacy of drug in relation to its safety. ¾ Pharmacokinetics of a drug should be investigated in patients because they may handle it differently from healthy people. Variation may occur due to the following reasons: •

Effect of disease

•

Age as compared with that of the volunteers studies that in phase-1 trials.

No. of Patients: We recruited hundred of patients for the conformation of phase-1 trials. Duration: It took 1-2 years for its completion.

A STUDY TO EVALUATE THE PHARMACOKINETICS AND SAFETY OF LEVOFLOXACIN IN PATIENTS WITH VARYING DEGREES OF RENAL FUNCTION PURPOSE: The primary objective was to evaluate the pharmacokinetics and safety of two dosing regimens of Levofloxacin in patients with varying degrees of renal function.

Condition

Intervention

Phase

Renal Diseases

Drug: Levofloxacin

Phase 2

Study Type

Interventional

Study Design

Treatment, Randomized, Open Label, Parallel Assignment, Pharmacokinetics Study

Official Title

An Open-Label Randomized Multiple-Dose Study to Evaluate Levofloxacin Steady-State Pharmacokinetics and Safety in Subjects With Varying Degrees of Renal Function

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FURTHER STUDY DETAILS AS PROVIDED Primary Outcome Measures: Evaluation of the pharmacokinetics of two dosing regimens of Levofloxacin in renally impaired and dialysis patients. Secondary Outcome Measures: Safety of two dosing regimens of Levofloxacin in renally impaired and dialysis patients. Estimated Enrollment:

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Study Start Date:

October 2007

Study Completion Date:

April 2009

Detailed Description: In this multiple-dose study conducted at 4 centers, the pharmacokinetics of two dosing regimens of levofloxacin were assessed in medically stable men and women with varying degree of renal function. The study consisted of a 21 day pretreatment screening phase, a 7-day open label treatment phase, and a 7 day posttreatment phase (or a follow-up phase for subjects with early study withdrawal). Patients were randomized into 1 of 10 treatment groups, for a total of 6 patients per group, based on degree of renal function to ensure that creatinine clearance values within each group represented the full range of values defined in the Food and Drug Administration's (FDA) 1998 guideline for pharmacokinetic studies in patients with impaired renal function. Fifty-nine patients were enrolled in the study. All patients received a single 750-mg dose of levofloxacin on Day 1; subsequent doses of either 250, 500, or 750 mg of levofloxacin (q24h or q48h) were based on renal function. Blood samples were collected from each patient from Day 1 to Day 14 for pharmacokinetic evaluation. Urine was collected on Days 1 and 7 before dosing and over specific time intervals up to 24 or 48 hours postdosing depending on the patient's dosing regimen. Dialysate samples were collected on Day 7 from HD patients immediately before dosing (as

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dialysis began) and at the end of the dialysis treatment. Patients were confined overnight at the study unit on Days 0, 1, 6, and 7, and remained confined until the 24 hour blood samples were collected on Days 2 and 8. Safety was based on the incidence, relationship to therapy, and severity of treatment-emergent adverse events and on changes in clinical laboratory values (hematology, chemistry, and urinalysis), vital sign measurements, electrocardiograms (ECGs), and physical examination findings. Single 750-mg dose of levofloxacin on Day 1; subsequent doses of Levofloxacin 250 milligram (mg), 500 mg, and 750 mg tablets administered every 24 hours for 7 days or every 48 hours for 7 days Eligibility Criteria: Ages Eligible for Study: 18 Years to 65 Years Genders Eligible for Study: Both Accepts Healthy Volunteers: No Inclusion Criteria: ¾ BMI between 18 and 35 kg/m2 ¾ No prescription or over-the-counter medications for previous 7 days ¾ Negative tests for drug and alcohol abuse, HIV, hepatitis B and hepatitis C ¾ Medically stable based on medical history, physical examination, 12-lead electrocardiograms, toxicology, antigen, and antibody screens, and clinical laboratory evaluations ¾ Stable renal function based on calculated creatine clearance for non-dialysis patients and the same dialysis treatment for at least 6 months prior to screening for dialysis patients ¾ Patients with creatinine clearance ≤80 mL/min who require treatment for renal impairment or other chronic disease (e.g., well-controlled diabetes, hypertension) must be on a stable treatment plan (medicines, doses, and regimens) for at least 2 months prior to Day 1 and during the entire study ¾ Hematocrit (hct) within the normal range based on patients' renal function at screening.

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Exclusion Criteria: ¾ Allergic reaction to quinolones ¾ Known or suspected allergy to heparin ¾ Clinically significant ECG or clinical laboratory abnormalities ¾ Creatinine clearance = 80 mL/min who required concomitant medication during the study ¾ Poorly controlled type 1 or type 2 diabetes ¾ Patients with creatinine clearance >= 50 mL/min with screening blood pressure outside the normal range (sitting systolic blood pressure 140 mm mercury [Hg] or diastolic blood pressure 90 mm Hg) ¾ Patients with CLCR