Workshop on GMP and Quality Assurance of Multisource Tuberculosis Medicines

Kuala Lumpur Malaysia 21-25 February 2005

Active Pharmaceutical Ingredients (APIs)

Theo Dekker, D.Sc., consultant to WHO Research Institute for Industrial Pharmacy North-West University, Potchefstroom, South Africa iiftgd@puk.ac.za
Feb 2005

TG Dekker WHO, Malaysia

Abbreviations
API BP CEP EOI FDC FPP GMP ICH Int.Ph. Ph.Eur. SmPC TB USP
Feb 2005

Active pharmaceutical ingredient British Pharmacopoeia EU certificate of suitability Expression of interest Fixed dose combination Finished pharmaceutical product Good manufacturing practices International Conference on Harmonization International Pharmacopoeia European Pharmacopoeia Summary of product characteristics Tuberculosis United States Pharmacopeia
2
TG Dekker WHO, Malaysia

Presentation approach
1. Collect and interpret all available information on the APIs (pre-dossier studies):
        
Feb 2005

The possible manufacturer(s) Literature, all aspects Monographs in pharmacopoeia Nomenclature Properties Manufacturing and site Specifications Container closure Stability testing / re-test period
3
TG Dekker WHO, Malaysia

2. Dossier requirements

Part 1. Available information on API

 Collecting and interpretation of all available information of the API through a systematic approach, should always be done upfront. Some outcomes:
1. Sound choice of API source (manufacturer) 2. Sound scientific understanding of the API, with respect to properties, stability, specifications, etc. 3. Assists in FPP pharmaceutical development 4. Assists in dossier compilation 5. Reduction of time / cost

 Forms part of Product Profile Report

Feb 2005

TG Dekker WHO, Malaysia

Info on potential API manufacturer(s)

The decision on the API manufacturer should be finalised before starting or early during FPP development studies
 Changes afterwards may be major of nature

1. Is the manufacturer reliable / reputable? 2. Is the open part of DMF available and according to all requirements? 3. Is a valid CEP available? 4. GMP inspection of API site by FPP manufacturer
Feb 2005

TG Dekker WHO, Malaysia

DMFs accepted & CEPs by Jan 2005

Number of manufacturers API Ethambutol 2HCl Isoniazid Pyrazinamide Rifampicin Streptomycin Total
Feb 2005

DMF 2 3 3 5 1 14
6

CEP 1 1 3

5
TG Dekker WHO, Malaysia

Literature information
Know your API before development, through:  Standard works / series / books such as:
    (Analytical) Profiles of Drug Substances and Excipients [ed: (Florey) Brittain) 30 volumes] The Merck Index (for structures, properties) Pharmaceutical Codex (12th edition) International Pharmaceutical Abstracts, Chemical Abstracts, Analytical Abstracts & internet

 Journals through search facilities such as

 Pharmacopoeial monographs (current)  Analysis of structure & stereochemistry

Feb 2005

TG Dekker WHO, Malaysia

Feb 2005

TG Dekker WHO, Malaysia

Examples of existing API information in standard works

 In the table on the next pages it is indicated in which standard works the APIs, appearing in the 5th invitation for Expression of Interest (TB), is included. Abbreviations:
 Apr = Analytical Profiles of Drug Substances and Excipients (contains chapters on APIs)  BP = British Pharmacopoeia  Cod = Pharmaceutical Codex (12th ed, 1994)  EP = Ph.Eur.  Int = International Pharmacopoeia  US = USP  MI = Merck Index (13th ed, 2001)
Feb 2005

TG Dekker WHO, Malaysia

Table of API occurrence

API Rifampicin (rifampin) Ethambutol 2HCl Pyrazinamide Isoniazid Streptomycin sulfate Amikacin Kanamycin Capreomycin
Feb 2005

APr Cod BP EP US

Int

MI

10

TG Dekker WHO, Malaysia

Table of API occurrence (con.)

API Cycloserine Ethionamide Ofloxacin Protionamide p-Aminosalicylic acid Moxifloxacin Apology: JP was not available at time of preparation
11

APr Cod BP EP US Int

MI

Feb 2005

TG Dekker WHO, Malaysia

Example: solubility from 2 literature sources

API
Rifampicin

Water
Water: Slightly 1,2 pH 7.5: 0.3% 2 pH 5.3: 0.4% 2 pH 2.0: 10% 2 50% 2 Sparingly 2 14% 1 1.5% 1
2

CHCl3
Freely 1,2

Ethanol
Slightly 2

Ethambutol 2HCl Ethambutol base Isoniazid Pyrazinamide

1

0.1% 2 Very 2 0.1% 1 0.7% 1

20% 2 2% 1 0.6% 2

Merck Index 13th ed

Pharmaceutical Codex 12th ed

TG Dekker WHO, Malaysia

Feb 2005

12

Information from the structures

 APIs which are organic compounds, have unique chemical structures & stereochemistry  These structures, together with the solid/liquid state conditions, are basically responsible for chemical and physical properties of the APIs  It is thus always appropriate to analyse the structure of the API, especially if limited literature information is available  Few examples to follow
Feb 2005

13

TG Dekker WHO, Malaysia

Rifampicin structure
hydrolysis

oxidation

hydrolysis

Feb 2005

14

TG Dekker WHO, Malaysia

3-Formyl rifamycin formation

Source: S. Singh et al. Pharm. Pharmacol. Commun., 6, 405-410 (2000)

Feb 2005

15

TG Dekker WHO, Malaysia

Rifampicin structure and properties

Oxidation  Hydroquinone group
 Main degradation of API (to rifampicin quinone)  Enhances solubility in alkaline medium

 Tertiary amine
 Moderately prone towards oxidation (to N-oxide)  Enhances solubility in acid medium

 Oxidation enhanced by
 Metal ions  Low pH

Feb 2005

16

TG Dekker WHO, Malaysia

Rifampicin structure and properties (2)

Hydrolysis  Hydrazone (imine) group
 Hydrolysis to 3-formyl rifamycin

 25-acetyl (ester) group

 Hydrolysis to 25-desacetyl rifampicin (minor)

Light sensitive
 Due to conjugation in molecule (unsaturated)

Storage of bulk raw material (BP/Ph.Eur.):

 Store under nitrogen in an airtight container, protected from light at temperature of 25C
Feb 2005

17

TG Dekker WHO, Malaysia

Rifampicin impurities (TLC)

 Reference: Int.Ph. related substances test for rifampicin Silica gel R1 CHCl3/methanol : 85/15 Daylight detection  BP limits for capsules:a) Rifampicin: 20 mg/ml b) Quinone: 0.8 mg/ml (4.0%) c) N-oxide:0.3 mg/ml (1.5%) d) 3-Formylrifamycin: 0.1 mg/ml (0.5%) e) Rifampicin: 0.2 mg/ml (1.0%)
Feb 2005

18

TG Dekker WHO, Malaysia

Isoniazid structure

Small molecule (quite stable)

 Basic amino functions  Primary amine - react with aldehydes/lactose (see presentation: Pharmaceutical R&D Considerations)  Can hydrolyze under stress conditions to inter alia isonicotinic acid & hydrazine  Oxidize in presence of strong oxidants (e.g. permanganate), with metals as catalyst 19

Feb 2005

TG Dekker WHO, Malaysia

Pyrazinamide structure

Small molecule (quite stable)

 Basic amino functions (in aromatic ring)  Amide group can hydrolyse under strong conditions to pyrazinoic acid & ammonia  USP ID test C: Boil 20 mg with 5 ml of 5 N sodium hydroxide: the odor of ammonia is perceptible  Forms metal complexes (slight pink bulk API / product?)  Sublimes when heated
Feb 2005

20

TG Dekker WHO, Malaysia

Pyrazinamide synthesis

 Reagents: NaOH, water as solvent, touch of EDTA

 Product directly crystallised from the reaction mixture  No residual solvents (only water used)  2-CPZ to be included as a possible synthesis impurity in API specifications  EDTA for metal complexation (prevent colouration)
Feb 2005

21

TG Dekker WHO, Malaysia

Ethambutol hydrochloride structure

Small molecule
 Basic amino groups (in free base)  No vulnerable groups for degradation under mild conditions (2-aminobutanol synthesis impurity)  2 chiral carbon atoms, optically active (test)  Hygroscopic (solubility in water: 50% m/m)  Can dissolve in absorbed water at high relative humidity  Forms metal complexes  USP: Preserve in well-closed containers
Feb 2005

22

TG Dekker WHO, Malaysia

p-Aminosalicylic acid

+ CO2
        Carboxylic acid and phenolic group: acidic Weak basic group (amphoteric) Saturated solution: pH of 3.0-3.5 (USP) Sodium salt available (monograph in USP) Labile: Decarboxylate when heated Limit test for m-aminophenol in USP (API & tablets) More stable in alkaline medium than in acid medium Store in cool place!! 23
TG Dekker WHO, Malaysia

Feb 2005

Cycloserine
R-configuration

 Optically active       

H20

Stable in anhydrous solid state, protected from water Degrades in solution, or when solid is exposed to moisture Pathway1: Dimerisation through one molecule attacking other Pathway 2: Hydrolysis to -aminoxy-D-alanine Stability: alkaline medium > neutral >> acid medium Dissolution medium capsules: buffer pH 6.8 !!! (USP) Primary amine: react with aldehydes/ketones 24
TG Dekker WHO, Malaysia

Feb 2005

Ofloxacin

Moxifloxacin

 Structurally related as encircled (see also ciprofloxacin)  Both APIs contain acid and basic groups  Chirality: both intrinsic chiral (optically active)  Ofloxacin: 1 chiral centre: racemate ( ) used  Moxifloxacin: 2 chiral centra: S,S-enatiomer used  Both APIs have enone system (in circle): photosensitive?
Feb 2005

25

TG Dekker WHO, Malaysia

Amikacin semi synthetic

Kanamycin
APIs differ only here

4 x NH2 groups, 2H2SO4 salt

hydrolysis

Feb 2005

26

TG Dekker WHO, Malaysia

Literature information sources

The information on the APIs mentioned in the previous slides are backed by inter alia:
 Analytical Profiles of Drug Substances and Excipients (ed: Florey/ Brittain) (see next slide)  The Pharmaceutical Codex: Principles and Practice of Pharmaceutics. 1994. Lund, W., ed. 12th edition, London: The Pharmaceutical Press  The Merck Index, 13th edition (2001)

Feb 2005

27

TG Dekker WHO, Malaysia

Literature support style

Literature information used in the dossier should always be accompanied by  Full traceable reference citations, for instance:
 Devani, M.B., Shishoo, C.J., Doshi, K.J. & Patel, H.B. Kinetic studies of the interaction between isoniazid and reducing sugars. Journal of Pharmaceutical Sciences, 74, 427-432 (1985)  Hassan, M.M.A., Jado, A.I., & Zubair, M.U. Aminosalicylic acid. In Florey, K., ed. Analytical Profiles of Drug Substances, vol. 10. New York: Academic Press, p. 1-27 (1981)

 Photocopies of the relevant pages

Feb 2005

28

TG Dekker WHO, Malaysia

Part 2. Dossier requirements for Active pharmaceutical ingredient (API)

Refer to Section 2 of: Guideline on Submission of Documentation for Prequalification of Multi-source (Generic) Finished Pharmaceutical Products (FPPs) used in the Treatment of HIV/AIDS, Malaria and Tuberculosis (hand-out) As from page 3/33
Feb 2005

29

TG Dekker WHO, Malaysia

Part 2. Dossier requirements for Active pharmaceutical ingredient (API) 2.1 2.2 2.3 2.4 2.5 2.6 2.7
-

3/33

Nomenclature (INN, Systematic, CAS, etc.) Properties (structure, stereochemistry, etc) Site of manufacture Route of synthesis (impurities, etc) Specifications (pharmacopoeia?) Container closure system Stability testing re-test period & storage
 Open part of Drug Master File - submit (DMF)  CEP (only limited information required)

Feb 2005

30

TG Dekker WHO, Malaysia

2.2 Properties of APIs 3/33

Three aspects: 2.2.1 API not described in BP, Int.Ph., JP, Ph.Eur., or USP (non-compendial) 2.2.2 API described in BP, Int.Ph., JP, Ph.Eur., or USP (compendial) 2.2.3 Information from literature (discussed)
 All the APIs on 5th Invitation for EOI fall in category 2.2.2, except moxifloxacin
Feb 2005

31

TG Dekker WHO, Malaysia

2.2.1 Properties non-compendial APIs

 Structure, stereochemistry, MF and RMM  Proof of structure/stereochemistry correctness
 Single crystal X-ray structure (sufficient) or  Spectrometric data (IR, 1H & 13C NMR, MS, etc.) in form of QA certified copies of the spectra and tabulated of data with - assignments against structure or - correlation against API spectral data from peer reviewed literature, preferable by the innovator (in tabulated form!!). Strongly recommended for non-compendial APIs

 Physico-chemical properties as discussed on next slide

Feb 2005

32

TG Dekker WHO, Malaysia

2.2.2 Properties compendial APIs

 Physicochemical and other relevant properties of the API, such as
 Solubility in water, other solvents such as ether, ethanol, acetone, and buffers of different pH  pKa, partition coefficient  Existence/absence of polymorphs and pseudo-polymorphs e.g. solvates (with XRPD, DSC, IR)  Polymorphism: see presentation Pharmaceutical R&D Considerations (rifampicin)  Hygroscopicity: see presentation Pharmaceutical R&D Considerations (ethambutol hydrochloride in 4FDC tablet)  Particle size
Feb 2005

33

TG Dekker WHO, Malaysia

2.3 Sites of manufacture 3/33

For each facility where all/part of manufacturing occurs:
     Name of manufacturer Street address Postal address Phone & fax numbers E-mail addresses

Feb 2005

34

TG Dekker WHO, Malaysia

2.4 Route(s) of synthesis 4/33

Three aspects: 2.4.1 API not described in BP, Int.Ph., JP, Ph.Eur., or USP (non-compendial) 2.4.2 API described in BP, Int.Ph., JP, Ph.Eur., or USP (compendial) 2.4.3 Specifications of raw materials and intermediates used in the synthesis

Feb 2005

35

TG Dekker WHO, Malaysia

2.4.1 Synthesis non-compendial APIs

 A flow diagram of the synthesis process, that includes inter alia chemical structures of starting materials and intermediates, reagents, catalysts, conditions, solvents and purification steps
 Example: see pyrazinamide synthesis (slide 21)

 A full description of each process, including purification and reprocessing (justified)  (Possible) process impurities should be discussed:
 Organic compounds  Catalysts and other inorganic impurities  Residual solvents
Feb 2005

36

TG Dekker WHO, Malaysia

2.4.2 Synthesis compendial APIs

 Valid CEP available
 Only outline of synthesis necessary

 No CEP
 Same as for non-compendial APIs

Feb 2005

37

TG Dekker WHO, Malaysia

2.5 API specifications 4/33

2.5.1 API not described in BP, Int.Ph., JP, Ph.Eur., or USP (non-compendial) 2.5.2 API described in BP, Int.Ph., JP, Ph.Eur., or USP (compendial) General note An API has only one set of specifications, applicable at release and throughout the retest period (FPPs may have two sets of specifications)
Feb 2005

38

TG Dekker WHO, Malaysia

2.5.1 Specs: non-compendial APIs

ICH Q6A (new APIs and products) for instance:
 Provide justification for proposed specifications  Impurities (synthesis, degradation & residual solvents) to be characterised and limits set  Analytical methods with validation  Preparation and potency determination/specification of primary and secondary (working) standards, with CoAs

 Valid CoAs for at least 2 batches

Feb 2005

39

TG Dekker WHO, Malaysia

2.5.2 Specs: compendial APIs

 Additional critical specifications not included in monograph e.g.
 particle size & polymorphic form  synthesis related impurities  residual solvents

 Valid CoAs for at least 2 batches 2.5.3 Container-closure system for API 4/33
 see guideline
Feb 2005

40

TG Dekker WHO, Malaysia

2.7 Stability testing 6/33

2.7.1 Stress testing (forced degradation) 2.7.2 Stability testing (regulatory)

Feb 2005

41

TG Dekker WHO, Malaysia

2.7.1 Forced degradation

 Literature information and/or CEP in support or to replace studies  Forced degradation studies
 To identify possible degradants for stability studies  To verify specificity of stability assay method - Diode array detection for API peak purity!!  Different stress conditions in solution (guideline)  Different stress conditions in solid state (guideline)
Feb 2005

42

TG Dekker WHO, Malaysia

2.7.2 Stability testing (regulatory)

 Only degradants that form under the real-time and accelerated conditions needs to be considered  Stability protocol, particulars of batches (3), tabulated results and discussion of data  Typical schedule given under FPPs  Propose re-test period when stored under defined conditions

Feb 2005

43

TG Dekker WHO, Malaysia

Some conclusions
1. Get to know your API by
     Analysis of literature information Analysis of the structures / functional groups Lab studies, e.g. forced degradation, spectral data and physical data Considering the dossier requirements DMF quality, GMP inspection, CEP availability

2. Decision on API manufacturer should include 3. API manufacturers are encouraged to apply for CEPs for their APIs
Feb 2005

44

TG Dekker WHO, Malaysia