Gunning for growth with GLP

By On Sep 30, 2013

Aruna Deshmukh

Need of Good Laboratory Practice (GLP) or similar organisational process, was always felt in the industry, for an assurance of quality. In the absence of any such process and due to commercial greed, products of questionable quality reached the markets and caused adverse effects or even deaths in humans. Since the early twentieth century, efforts were being made to put Government regulations in place, to assure quality and safety of products, before their marketing approval for human consumption. Needless to say that, these earlier regulations were weak and with in-built flaws. Therefore advantages were taken by the industries to make profits by releasing products for sale without, adequate verification of their quality or safety. Unfortunately most regulations were promulgated after the tragedies had occurred. Finally in June 1979, the US FDA implemented GLPs in the US. In spite of it, in the US itself as recently as in 2001, a case of serious violations was detected by the US FDA and it had issued a warning letter of disqualification to ‘Coulston Foundation’, because during its facility inspection by FDA, serious violations/flaws were detected in data integrity, which would have had wide spread consequences for human safety (Lepay, 1999). Even now we continue to encounter adverse events. For example, Cerivastatin and Rosuvastatin were withdrawn from the market due to Rhabdomyolysis. COX-2 inhibitors have shown cardiac and hepatic toxicity. These examples, beyond doubt reiterate that, there is a ‘Need of GLP in Industry’ for absolute assurance of quality and safety of products, particularly in post-GATT era of Global competition, when each industry is fiercely competing to bring its products ‘fast and first into market’.

History

“We can never be fully in possession of a science until we know the history of its development” – Charles Greene Cumston, 1926.

The making of GLP regulations was preceded by several tragic events, where human life was either lost or compromised. Finally enforcement of the GLP regulations in 1979, was culmination of several such events. Although, it is not the complete list, following are just a few examples of the tragic events that occurred in the last 100 years.

1901 (US): Diphtheria antitoxin got contaminated with tetanus and 130 children died.
1930 (EU): In Germany, BCG vaccine was contaminated causing 75 deaths and 160 injuries.
1930 (US): Lyndol oil was used in ‘Jamaican ginger’ resulted in spinal cord injury, causing irreversible paralysis to over 50,000 people.
1937 (US): Sulfanilamide elixir, containing diethylene glycol, caused 107 deaths by renal toxicity, mostly children, due to commercial release of untested drug by M/s SE Massengill Co, in Bristol, Tennessee, (US). Company’s chief chemist, Harold Cole Watkins, who had prepared the ‘elixir’ committed suicide after learning of the effects of his latest concoction.

In 1959, ‘Grey baby syndrome’ was reported due to Chloramphenicol toxicity responsible for deaths in new born babies even at therapeutic doses (Sutherland, 1959).

Between 1956-61, ‘Thalidomide tragedy’ shook entire world and scientific community. Thalidomide was introduced in Europe, by a West German pharmaceutical company, Chemie Gruenthal, for the treatment of nausea in early pregnancy and as a mild sedative. It was sold as an OTC product by 1957 in Europe. About 12,000 children were born with phocomelia and many others with eye and ear defects (McBride, 1961). Thalidomide was never allowed to reach the US market. Thanks to the single handed efforts made by one lady, Dr Frances Kelsey, of US FDA (2001). She refused to grant permission to sell Thalidomide in US market and thus became ‘Bete Noire’ of M/s Richardson-Merrell Pharmaceutical Co, who had applied to FDA for permission to sell Thalidomide (named as Kevadon) in US. After the ‘Thalidomide tragedy’, several amendments in drug safety acts in US (Kefauver-Harris amendment, 1962) and EU were made requiring the pharma industries to give evidence of safety and even efficacy before marketing a new drug.

Principles of GLP

During the year 1975, US FDA inspected Searle Pharmaceuticals and Industrial Bio-Test Laboratories to review and monitor the preclinical data submitted. Several examples of irregularities and fraudulent practices were noticed. A few are given below (Dent, 2004).

Results were found to be manipulated to suit the requirements, scientific findings were faked or suppressed, animals were given wrong doses or even wrong compounds, animals found dead on the previous page of the register, became alive on the next page and were found to be merrily living till end of the study, entire carcass of animals were preserved in fixatives and histopathology reports were submitted without microscopic examination of tissues.

During FDA inspections, it had revealed many cases of badly managed studies by unqualified and untrained study directors and study personnel. The protocols were poorly designed and procedures were not followed properly. The equipment were not calibrated. Characterisation of test items and test systems and archiving were inadequate. There were discrepancies or no traceability or approval of raw data by responsible study personnel. Standard procedures were not laid down and animal husbandry was poor.

In 1975-76, as a fall out of these inspections, the US FDA in collaboration with Searle Pharmaceuticals, drafted first GLP proposal and published it in the Federal Register on 19th November 1976. Final draft rule 21CFR part 58 was published on December 22, 1978. It became a law on June 20, 1979. Final rule for industries was published in 1987 as ‘Guidance for Industry’ and it was last revised in 1997.

In 1981 Organisation for Economic Cooperation and Development (OECD) adopted GLP guidelines, which were revised in 1997, cancelling and replacing the original principles adopted in 1981 (OECD, 1998). International Conference on Harmonization (ICH) was formed to harmonise GLP guidelines in Europe, Japan and the US (ICH, 1990). On November 26, 1997, Mutual Acceptance of Data (MAD) agreement was signed (OECD, 1997). The principle of GLPs has been to promote development of quality test-data. Comparable quality of test-data across the industries and countries, formed the basis of ‘MAD’ agreement.

In India, National GLP Compliance Monitoring Authority was established by the Department of Science and Technology, Government of India, with the approval of the Union Cabinet on April 24, 2002. Presently, India enjoys the status of a provisional member of the OECD for GLP. India is an observer to the OECD’s Working Group on GLP and also a member of the OECD Test Guidelines Programme (DST 2005).

Since the theme of this chapter is ‘Need of GLP in Industry’, it will be prudent to take a look at the short overview of the principles of GLPs to understand its ‘need’ in the industry. Through the salient features of this overview (US FDA 1997, OECD 1998 and OGLP 2000), we shall be able to appreciate that ‘GLPs’ have been prepared as ‘Guidance to Industries’ to assure quality of preclinical safety data. Needless to say, that the procedures described in these ‘guidance documents’ have been designed, especially for the pharma industries, which are engaged to discover, test, manufacture and market new drugs.

Scope: The principles of good laboratory practices are applied to the testing of chemicals to obtain data on their properties and/or their safety with respect to human health or the environment. It also includes the work conducted in the field studies and these data are developed for the purpose of meeting the regulatory requirements.
Definitions of Terms

GLP: It is a quality system concerned with the organisational process and the conditions under which non-clinical health and environmental safety studies are planned, performed, monitored, recorded, archived and reported.
Please note the underlined words in the above mentioned definition of GLP. These words have specific meaning and refer to the respective functions, covered under GLPs. Any industry, aiming to be a GLP compliant facility, needs to address each issue, that has been elaborated in the following paragraphs.
Study categories: These are toxicology, mutagenicity, ecotoxicology, environmental behaviour and bio-accumulation, residue analysis, impact on mesocosms and natural ecosystems, physical and chemical properties and analytical chemistry.
Test facility: It means the person/s, premises and operational unit/s, required for carrying out non-clinical safety studies.
Inspection: It is undertaken to ascertain that the test facility carries out processes and methods and data generated comply to GLP principles.
Test facility management: The person/s having authority and responsibility for the functioning of test facility according to the principles of GLP.
Sponsor: The entity, which commissions or supports conduct of non-clinical safety studies.
Study director: The individual responsible for overall conduct of non-clinical safety study.
Principle investigator: Acts on behalf of the study director for conduct of multi-site studies.
Quality assurance programme: Test facility should have a documented quality assurance programme to ensure that studies performed are in compliance with the principles of GLP.
Standard operating procedures (SOPs): These are documented procedures describing about how to perform test or activities normally not specified in study plans or guidelines.
Study plan: For each study, a written plan should exist prior to the initiation of study. It defines the objective and experimental design for the conduct of the study. It should be approved by dated signature of study director and verified by quality assurance personnel. The study plan should also be approved by the test facility management.
Master schedule: It is a compilation of information regarding all the ongoing studies in the test facility. It helps for the assessment of work-load of study personnel and for the tracking of studies at the test facility.
Test system: Any biological, chemical or physical system/agent or a combination thereof.
Raw data: All original test facility records and documentation or verified copies thereof depicting results of original observations and activities in a study.
Specimen: Any material derived from a test system for examination, analysis and retention.
Experimental starting date: Date on which first study specific data are collected/recorded.
Experimental completion date: The last date on which data are collected from the study.
Study initiation date: The date on which study director signs the study plan.
Study completion date: It means the date study director signs the final report.
Test item: It is a substance (chemical or a mixture) which is under investigation.
Reference/control item: Any article used to provide a basis for comparison with test item.
Vehicle: Any agent serving as a carrier employed to mix, disperse or solubilise the test or reference item to facilitate administration / application to the test system.

Organisation and personnel

Responsibilities of management: The test facility management has the authority and formal responsibility for the functioning of the test facility according to the principles of GLPs. It should enroll qualified, experienced and trained study directors and study personnel before initiation of the study. Provide appropriate facilities, equipment and materials for proper conduct of each study. The functions of each person should be clearly defined and where necessary training is given and its records are kept. It should put in place SOPs and a quality assurance programme with properly trained personnel. The management should also provide safety and health monitoring systems, for all study personnel, according to national/international regulations.
Responsibilities of the study director: Study director is the single point of study control. He has responsibility for overall conduct of the study. He should prepare study plans and get approval from the management, before each study start. He is responsible to document and record the raw data generated during the study as per study plan. At termination of the study, he should prepare final report and get it approved from quality assurance. The study director must archive the study plans, final reports, raw data and any other supporting material related to the study, at the termination of each study.
Responsibilities of personnel: All the study personnel should have adequate qualifications and training for conduct of study, noting down clinical observations and raw data recording. They should exercise safe working practice as per SOPs and should take proper health precautions to minimise risks to themselves and the test system for integrity of the study. Personnel having illness or any medical condition, that is likely to have any adverse effect on the study, should be excluded from the operations of that study until recovered medically.

Test system

It is any biological, chemical or physical system/agent or a combination thereof used in a study. Suitable conditions are required to be established for maintenance and care of biological test system. Newly received animals/plant test system should be isolated until their health status has been evaluated. At the experimental starting date the test system should be free of any diseases, which might interfere with conclusion of the study. Records of its source, date of arrival and conditions at arrival, should be maintained. Biological test system should be acclimatised to the test environment for an adequate period before administration of test item for the first time. All information needed to properly identify the test system should appear on the labels of their housing or containers.

Test and control items

Receipt, handling, sampling and storage: All the records regarding test item characterisation, date of receipt, quantities received and used in the study, should be maintained. Handling, sampling and storage procedures should be identified in order to ensure homogeneity and stability and to prevent contamination or mix-ups. Storage container should carry identification number, expiry date and specific storage condition/s.
Characterisation: Each test/reference item should be appropriately identified (CAS number, name). For each study, identity, batch number, purity, stability, composition, concentration or other characterisations should be defined for every batch with respect to test and control items. If the test item is administered in any vehicle, the procedure/s should be established for testing the homogeneity and stability of test item in that vehicle. A sample for analytical purpose from each batch of test item should be retained for studies in which the test item is tested longer than four weeks.

Facilities and equipment

Facilities: The test facilities of a suitable size, construction and location should be made available to meet the requirements of the study. Attention should be paid to minimise disturbances that would interfere with validity of the study. Design of the test facility should provide an adequate degree of separation of different activities to assure proper conduct of each study. It should have separate facility to house different test systems, individual projects and sufficient number of rooms having controlled temperature and humidity as specified (Guide for the care and use of laboratory animals, 1996). To prevent contamination and mix ups, separate rooms for receipt and storage of test and reference items and for mixing the test items with vehicles should be provided. Archiving facility of adequate size with appropriate separation for storage and retrieval of raw data, reports, samples, wet and dry specimens should be made available. Archives should have proper security and SOPs for its functions. Handling and disposal of wastes generated during performance of study should be carried out in a manner consistent with national/international regulatory requirements.
Equipment: Equipment / apparatus of proper design and capacity used for the generation, storage and retrieval of data and for controlling environmental factors relevant to the study and validated computerised systems, should be suitably located. Any equipment used in the study should be periodically inspected, cleaned, maintained and calibrated according to SOPs. Record of these activities should be maintained.

Standard Operating Procedures (SOPs)

General: SOP is a document, described as ‘Write what you do and do what you write’. A test facility should have written SOPs approved by the management which are intended to assure quality and integrity of the data generated in preclinical safety studies. Each laboratory unit should have immediately available SOPs relevant to the activities performed. Text books, articles and manuals may be used as supplement to the SOPs.
Application: SOP should be available for following categories of laboratory activities/functions:

Test/ reference items: Receipt, identification, labeling, handling, sampling, storage.
Equipment/ apparatus and reagents: Use, maintenance, cleaning, calibration, environmental control and preparation of reagents.
Record keeping, reporting, storage and retrieval: Coding of studies, data collection, preparation of reports, indexing systems, handling of data and computerised data.
Test system: Room preparation and its environmental conditions. Procedures for receipt, transfer, proper placement, identification and care of test system. Observations to be made before, during and at termination of the study. Handling of animals found moribund or dead during the study. Collection, identification and handling of specimens including necropsy and histopathology.

Performance of the study

Study plan: For each study, a study plan should exist in a written form prior to initiation of the study and it should be retained as raw data. All changes, modifications or revisions of the study plan should be documented, signed and dated by the study director and should be maintained with the study plan.
Content of the study plan: It should contain identification of the study, test and reference items and a descriptive title. A statement describing nature and purpose of the study. Date of agreement to study plan by signature of the study director and test facility management/sponsor. Proposed starting and completion dates. Reference of the test guidelines. Justification of test system and dosage. Method of administration and justification of its choice. Information regarding chronological procedures of the study, material and methods, type and frequency of analysis, observations and examinations to be performed.

Reporting and archiving

Reporting: A final report should be prepared for each study by the study director. The report should indicate extent of GLP compliance and validity of data. It should include identification of the study, test and reference items and a descriptive title. Information concerning the sponsor and the test facility and experimental starting and completion date. A quality assurance programme statement, listing the type of inspections made with dates and confirmation that the final report reflected the raw data. Description of material and test methods, details of the results including tables and determination of statistical significance, discussion and conclusion. Information regarding archiving of study plan, test and reference items, specimens, raw data, reports etc.
Archiving: Following should be retained in the archives for the period specified by the authorities. Study plans, raw data, samples of test and control items, wet specimens and final report of each study. Records of all inspections performed by quality assurance programme and master schedules. Records of qualifications, experience, training, job description and health monitoring of study-personnel. Records of maintenance and calibration of equipment. Validation documents of computerised system, historical file of all SOPs, environmental monitoring record etc. Materials retained in the archives should be indexed to facilitate orderly storage and retrieval. Only personnel authorised by the management should have assess to archives. Movement of material in and out of the archives should be properly documented.

Quality assurance programme

General: The test facility should have a documented quality assurance programme to ensure that the studies performed are in compliance with the principles of GLPs. It should be carried out as per SOPs by trained individual or individuals designated by and directly responsible to test facility management and who are familiar with the test procedures. The individual should not be involved in conduct of the study being assured/audited.
Responsibilities of the quality assurance (QA) personnel: Maintain copies of all approved study plans, SOPs and master schedule. QA should verify study plans for compliance with the principles of GLPs and this compliance should be documented. Inspections should be conducted to determine that the studies were carried out in accordance with the study plans and SOPs. The inspections could be study based, facility based and process based. Records of such inspections should be retained. Final report should be inspected for accurate reporting of results and complete reflection of raw data of the study. Results of inspections should be promptly reported in writing to the management and study director. A signed statement should be included in the final report specifying types of inspections made with dates. This statement would serve to confirm that the final report correctly reflected the raw data.

Epilogue

Events enumerated in the ‘History’ and ‘Introduction’ have proven beyond doubt that, regulatory guidelines were necessary for the industries, to establish proper systems to assure quality and safety of products before their marketing, to prevent man made disasters, due to negligence. After going through the overview of GLPs, it is quite evident that each step, therein, has been designed to address issues related to industries. The GLP ‘guidance documents’ are especially applicable to the pharma industries for assurance of quality, safety, truthfulness, validity and integrity of preclinical safety studies.

It is a system which enables to reconstruct the events of preclinical safety studies, such as how and where the studies were conducted, how the data were generated and archived, who was responsible, whether the raw-data and reports were audited, as per GLP regulations, by QA programme and a statement to the effect was issued by QA.

In each industry, ‘Test facility management’ is supposed to have an authority and overall responsibility to implement principles of GLPs, appoint study-directors and study personnel with appropriate qualifications, experience and training. Arrange training programmes and maintain records of such programmes.

In GLPs each step or process has been made auditable, such as characterisation of test and control items, their purity, stability, stability in vehicle, storage conditions etc. For test systems, methods for maintenance of culture for in-vitro systems and for in-vivo systems, proper housing conditions, adequate facilities with sufficient number of rooms for segregation of species and studies, specified temperature and humidity control in each room, clean and dirty areas, regular analysis of feed and water and documentation of records of reach activity to help the audit process during inspections.

In post-GATT scenario, every industry, especially pharma industry, is looking forward to be an international player and to achieve this goal, GLP compliance has become not only necessary but almost mandatory. India has shown great potentials in every scientific field. Just like in IT industry, India is all set to be a global player in pharma industry, as well, and to achieve that goal, they need to comply with the principles of GLPs.

Indian National GLP compliance monitoring authority has already taken a step forward in this direction and has commenced GLP inspections of the pharma industries, that carry out preclinical safety studies for filing of INDs and NDAs.

Indian pharma Industry, with GLP firmly in place, soon would be the ‘numero uno’ and would be able to prevent commercial release of improperly tested new drugs, that may be responsible for yet another ‘Thalidomide’ like tragedy.