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

The possible manufacturer(s)

Literature, all aspects
Monographs in pharmacopoeia

2. Dossier requirements

Feb 2005

Nomenclature
Properties
Manufacturing and site
Specifications
Container closure
Stability testing / re-test period
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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

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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
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DMFs accepted & CEPs by Jan 2005

Number of manufacturers
API

DMF

CEP

Ethambutol 2HCl

Isoniazid

Pyrazinamide

Rifampicin

Streptomycin

1
Total

Feb 2005

14
6

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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)

Journals through search facilities such as

International Pharmaceutical Abstracts, Chemical

Abstracts, Analytical Abstracts & internet

Pharmacopoeial monographs (current)

Analysis of structure & stereochemistry
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Feb 2005

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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)
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Table of API occurrence

API

APr Cod BP EP US

Int

MI

Rifampicin (rifampin)

Ethambutol 2HCl

Pyrazinamide

Isoniazid

Streptomycin sulfate

Amikacin

Kanamycin

Capreomycin
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Table of API occurrence (con.)

API
Cycloserine

APr Cod BP EP US Int

Ethionamide

Ofloxacin

Protionamide
p-Aminosalicylic acid

MI

Moxifloxacin

Apology: JP was not available at time of preparation

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Example: solubility from

2 literature sources
API

Water

CHCl3

Ethanol

Rifampicin

Water: Slightly 1,2 Freely 1,2

pH 7.5: 0.3% 2
pH 5.3: 0.4% 2
pH 2.0: 10% 2

Slightly 2

Ethambutol 2HCl
Ethambutol base
Isoniazid
Pyrazinamide

50% 2
Sparingly 2
14% 1
1.5% 1

20% 2

Merck Index 13th ed

Feb 2005

0.1% 2
Very 2
0.1% 1
0.7% 1

2% 1
0.6% 2

Pharmaceutical Codex 12th ed

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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
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Rifampicin structure
hydrolysis

oxidation

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hydrolysis

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3-Formyl rifamycin formation

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

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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
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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
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Rifampicin impurities (TLC)

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

a)
b)
c)
d)

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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
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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
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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)
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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
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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!!

Feb 2005

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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
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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?
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Amikacin semi synthetic

Kanamycin
APIs differ only here

4 x NH2 groups, 2H2SO4 salt

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hydrolysis

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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)

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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

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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
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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)
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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
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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
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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
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2.3 Sites of manufacture 3/33

For each facility where all/part of
manufacturing occurs:

Feb 2005

Name of manufacturer
Street address
Postal address
Phone & fax numbers
E-mail addresses

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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

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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
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2.4.2 Synthesis compendial APIs

Valid CEP available
Only outline of synthesis necessary

No CEP
Same as for non-compendial APIs

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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)
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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

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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
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2.7 Stability testing 6/33

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

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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)

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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

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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

2. Decision on API manufacturer should include

DMF quality, GMP inspection, CEP availability

3. API manufacturers are encouraged to apply

for CEPs for their APIs
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