Perubahan Sumber Bahan Baku

Zat Aktif Obat Wajib Uji BE

Coaching Clinic Pelaksanaan Uji Bioekivalensi

Bandung, 12 September 2022
Prof. Dr.rer.nat. apt. R. Emran Kartasasmita, M.Si.
Sekolah Farmasi ITB
 API
 Structure, Characteristics,
Sintesis / Mod. Reactivities, Biological Activities
Kimiawi  Keamanan, Khasiat, dan Mutu
Bahan Obat (API)
Pemula

Karakteristik Fisikokimia,
Reaktivitas, Kemurnian

Rute Fase Fase Fase H S

Farmasetik Farmakokinetik Farmakodinamik u y
Pemberian
m s
a t
n e
QC/QA Uji Uji m
Disolusi Bioekuivalen Aktivitas Biologi
= f (Kar. Fis.Kim)
Farmakope
X
Mutu Khasiat Ef. Farm. Ef. Toks. Keamanan
Pharmaceutical Application Complex Processes
phase of Drugs
Dissolution of
Active Compound

Pharmacokinetic
phase Resorpstion

Biotransformation
Deposition Distribution
Excretion
Target
Pharmacodynamic (Receptor)
phase
Pharmacological
Effects

Pharm. Effects Toxic Effects

Drug Transport and Drug Receptor
Interaction
 Bioequivalence -
General Considerations

Dr. John Gordon

6th Annual Prequalification Team - Medicines Quality Assessment Training

Copenhagen, May 18-21, 2014
 Establishing Equivalence
 Comparative pharmacokinetic studies
– In vivo comparative bioavailability studies
– Comparison of performance of Finished Pharmaceutical Products (FPPs)
based rate and extent of absorption of API from each formulation
• Area under the concentration-time curve (AUC)
• Maximal concentration (Cmax)
• Time to maximal concentration (Tmax)

 Comparative pharmacodynamic studies

 Comparative clinical trials
 Comparative in vitro methods
– Biopharmaceutics Classification System (BCS)-based biowaivers
– Additional strengths biowaivers

6th Annual Prequalification Team - Medicines Quality Assessment Training

6| Copenhagen, May 18-21, 2014
 Bioequivalence

 Finished Pharmaceutical Products (FPPs) are bioequivalent if

– they are pharmaceutically equivalent or pharmaceutical alternatives
– bioavailabilities (both rate and extent) after administration in the same molar dose
are similar to such a degree that their effects can be expected to be essentially
the same

 Pharmaceutical alternative
– Same molar amount of the same API(s) but differ in dosage form (e.g., tablets vs.
capsules), and/or chemical form (e.g., different salts, different esters)
– Deliver the same active moiety by the same route of administration

6th Annual Prequalification Team - Medicines Quality Assessment Training

7| Copenhagen, May 18-21, 2014
 Establishing Bioequivalence

FPPs being tested

Comparator product
– WHO provides recommendations
– To be discussed shortly

Test product
– Biobatch of sufficient size
– Consistent with product proposed for market
– To be discussed later

6th Annual Prequalification Team - Medicines Quality Assessment Training

8| Copenhagen, May 18-21, 2014
 Establishing Bioequivalence

Important PK parameters

Cmax:
the observed maximum concentration of a drug
 measure of the rate of absorption
AUC:

area under the concentration-time curve tmax:

 measure of the extent of absorption time at which Cmax is observed
 measure of the rate of absorption

6th Annual Prequalification Team - Medicines Quality Assessment Training

9| Copenhagen, May 18-21, 2014
 ASEAN GUIDELINES FOR THE CONDUCT OF BIOAVAILABILITY AND
BIOEQUIVALENCE STUDIES – QUESTIONS AND ANSWERS ( Q & A )
(Version 4)
Generally, a new BE study is not required for changes of API source. However, applicants should
provide comparative dissolution data depending on the dosage form of
the proposed product.
To substitute an API source, all of the following 2. The previous batch used in BE studies and the
circumstances should be complied: BE studies were acceptable, except for the
1. The previous API source is not being withdrawn cGMP issues that were specific to the previous
due to deficiencies specifically relating to that API.
API, such as: 3. The specifications of the API from new
 Lack of adequate controls source are essentially the same as the API
 Evidence of adulteration from previous source, such as:
 Evidence of falsification of data in the  The particle size,
application or identified in the preapproval  polymorphic form,
inspection.  no change in route of synthesis,
 impurities profile, etc.
 Particle Size
• Methods used to measure a particle size • d10, d50 and d90 in a PSD
distribution d10, d50 and d90 are so-called percentile
There are many methods to determine the values. They indicate the size below which 10%,
particle size distribution of a sample. Commonly 50% or 90% of all particles are found.
used methods are: • Monomodal and bimodal PSD
 sieve analysis, The mode size found where the frequency
 laser diffraction, distribution reaches a maximum:
 dynamic light scattering and image analysis.  One maximum: monomodal
• Importance of Particle Size Distribution  Two maxima: bimodal distribution
(PSD)  More maxima: multimodal
PSD is an important quality criterion for many • Specification of PSD
products, but also for raw materials. These The width of PSD an important statistical
include, for example: property. The width of the distribution can be
 flowability, surface area, conveying given, for example by:
properties,  the SD around the mean value (mean
 extraction and dissolution behavior, particle size) (note: FDA uses Median)
 reactivity,  or the SPAN value [ (d90-d10)/d50 ]
 abrasiveness and even taste.
Source: Molecules 2015, 20, 18759-18776; doi:10.3390/molecules201018759
 d10, d50, d90 of a PSD

Source: https://www.microtrac.com/knowledge/particle-size-distribution/#:~:text=What does d10%2C d50 and,of all particles are found. 3/18/22, 7:29 PM
 Monomodal and Bimodal PSD

Span:
 Monomodal: 0,436
 Bimodal: 0,557

Source: https://www.microtrac.com/knowledge/particle-size-distribution/#:~:text=What does d10%2C d50 and,of all particles are found. 3/18/22, 7:29 PM
 Polymorphic Form
• Crystalline polymorphs: the same chemical composition, but different internal
crystal structures
• It is quite common among organic molecules, and many drugs can crystallize
into different polymorphic forms:
 Different lattice structures and/or different molecular conformations
 Different physicochemical properties
• Generally, the solubility of metastable polymorphs is kinetically higher
than that of a thermodynamically more stable polymorph: can be a solution to
improve bioavailability

Source: Molecules 2015, 20, 18759-18776; doi:10.3390/molecules201018759

 Polymorphic Form

• Polymorphism is widely prevalent in pharmaceutical solids. About 50% of

known drug compounds are known to be polymorphic.
• Polymorphs are considered as the supramolecular isomers or super
isomers of a compound that differs in their molecular arrangements in the
supramolecule, that is, the crystal.
• These differences in molecular packing arrangements can have origins in the
molecular geometry (different conformations or tautomers) or primary
supramolecular recognition modes (different synthons) and are classified as
conformational, tautomeric, and synthon (synthetic building block)
polymorphs
Thakuria, R.; Thakur, T. S. (2017) Crystal Polymorphism in Pharmaceutical Science.
 Polymorphic Form

• a synthon is a hypothetical
unit within a target molecule
that represents a potential
starting reagent in the
retroactive synthesis of that
target molecule. The term was
coined in 1967 by E. J. Corey.
• In 1988 he noted that the
"word synthon has now come
to be used to mean
synthetic building
block rather than
retrosynthetic fragmentation
Thakuria, R.; Thakur, T. S. (2017) Crystal Polymorphism in Pharmaceutical Science. structures".
 Tautomer of Omeprazole

Rosa M. Claramunt, Concepción López, and José Elguero, The structure of Omeprazole in the solid state: a 13C and 15N
NMR/CPMAS study, ISSN 1424-6376, unpublished work.
Contoh Polymorph Pada Ranitidin
 Examples of Polymorphism of Several Drugs

Source: Molecules 2015, 20, 18759-18776; doi:10.3390/molecules201018759

 Examples of Polymorphism of Several Drugs

Source: Molecules 2015, 20, 18759-18776; doi:10.3390/molecules201018759

 Differences in Synthesis Route and Impurities
Konsekuensi Perbedaan Rute Sintesis
• Reaktan/starting material berbeda • Proses working-up produk akhir
• Reagent berbeda berbeda: perbedaan proses
• Tahap sintesis berbeda kristalisasi (pengaturan suhu,
pelarut, pengadukan) bisa
• By product/impurities berbeda menyebabkan polimorfisme
 Contoh Sintesis 1: Imatinib-1
• Imatinib mesylate is an anti-neoplastic (anti-cancer) drug used for the
treatment of chronic myeloid leukemia and gastrointestinal stromal tumor
• U.S. Pat. No. 5,521,184 and WO 03/066613 describe synthetic routes for preparing imatinib.
• One synthetic process, depicted in Scheme 1, involves:
• Reacting 4-(4-methyl-piperazin-l-ylmethyl)-benzoic acid methyl ester with 3-nitro-4- methyl-
aniline to obtain N-(4-methyl-3-nitrophenyl)-4-(4- methyl-piperazin-l-ylmethyl)-benzamide,
which is subsequently reduced to obtain N-(3-amino-4-methyl-phenyl)-4-(4-methyl-piperazin-l-
ylmethyl)-benizamide.
• The latter is reacted with cyanamide (NH2CN) in a mixture of
concentrated hydrochloric acid solution and n-butanol to produce N-(3-guanidino-4-methyl-
phenyl)-4-(4-methyl-piperazin-l-ylmethyl)-benzamide, which is subsequently reacted with 3-
dimethylamino-l-pyridin-3-yl-propenone to obtain imatinib.
Contoh Sintesis 1:
Imatinib-1
Contoh Sintesis 1: Imatinib-2
 Contoh Sintesis 2: Imatinib-1
• Another process, depicted in Scheme 2, involves reacting:
• 3-bromo-4-methyl-aniline with 4-(4-methyl-piperazin-1-ylmethyl)-benzoic acid
methyl ester to obtain N-(3-bromo-4-methyl-phenyl)-4-(4-methy1-piperazin-1-
ylmethyl)-benzamide.
• The latter is reacted with 4-(3-pyridyl)-2-pyrimidine amine (which is obtained
by reacting cyanamide with 3-dimethylamino-l-pyridin-3-yl-propenone) to
obtain imatinib.
Contoh Sintesis 2: Imatinib-2
Contoh Sintesis 2: Imatinib-3
 Contoh Sintesis 3: Imatinib-1
• Another process, depicted in Scheme 3, includes:
• Obtaining 2-methyl-5-nitrophenyl-guanidine from 2-amino-4-nitro-toluene by
adding nitric acid to a solution of the latter in ethanol followed by addition of
cyanamide.
• The product is subsequently reacted with 3-dimethylamino-l-pyridin-3-
ylpropenone to obtain N-(2-methyl-5-nitrophenyl)-4-(3-pyridyl)-2-pyrimidine-
amine, which is subsequently reduced and reacted with 4-(4-methyl-
piperazinomethyl)-benzoyl chloride to obtain imatinib.
Contoh Sintesis 3: Imatinib-2
Contoh Sintesis 3: Imatinib-3
Contoh Sintesis 4: Imatinib-1
 Contoh Sintesis 4: Imatinib-2
• The syntheses described in U.S. Pat. No. 5,521,184 and WO 03/066613 are not particularly
suitable for industrial purposes (Expensive reagents)
• US 7,550,591 B2: direct coupling of 4-(4-methyl-piperazin-l-ylmethyl)-benzoic acid with N-(5-
amino-2-methylphenyl)-4-(3-pyridyl)-2-pyrimidineamine in the presence of a carboxylic acid
coupling reagent, to produce imatinib, and optionally converting the imatinib into a salt.
• The carboxylic acid coupling reagent can include coupling reagents that are conventionally
used for activating and coupling carboxylic acids. A preferred class of carboxylic acid coupling
reagents are carbodiimide coupling reagents.
• Exemplary carbodiimide coupling reagents include N-(3-dimethylaminopropyl)-N-ethyl-
carbodiimide (EDC) and salts thereof, e.g., the hydrochloride salt (EDC HC1).
Contoh Sintesis 4: Imatinib-2
 Impurities Imatinib

• Banyak jenis impurities

• Impurity 1 – 9
• Impurity A – E
• Two process impurities:
• 4-[(4-Methyl-1-piperazinyl) methyl]benzoic acid dihydrochloride
(MPBA)
• 4-Methyl-N3-[4-(3-pyridinyl)-2-pyrimidinyl]-1,3-benzenediamine
(PNMP)
• Classified as potential genotoxic impurities
Maziarz, Margaret & Wrona, Mark. (2017). Analysis of Genotoxic Impurities of Imatinib Mesylate by LC-MS from Early Development to
Routine Monitoring. 10.13140/RG.2.2.32367.64168.
 Impurities Imatinib

Maziarz, Margaret & Wrona, Mark. (2017). Analysis of Genotoxic Impurities of Imatinib Mesylate by LC-MS from Early Development to
Routine Monitoring. 10.13140/RG.2.2.32367.64168.
 Impurities Imatinib

Maziarz, Margaret & Wrona, Mark. (2017). Analysis of Genotoxic Impurities of Imatinib Mesylate by LC-MS from Early Development to Routine Monitoring.
10.13140/RG.2.2.32367.64168.
 Impurities Imatinib

Maziarz, Margaret & Wrona, Mark. (2017). Analysis of Genotoxic Impurities of Imatinib Mesylate by LC-MS from Early Development to Routine Monitoring.
10.13140/RG.2.2.32367.64168.
Contoh Sintesis 5: FAVIPIRAVIR
 FAVIPIRAVIR SYNTHESIS

1st synthesis route in 2000 by Y. Furuta and coworkers

Overall yield = 0.44%

Furuta, Y.; Egawa, H. WO Patent 20000/10569, 2001-04-07

 FAVIPIRAVIR SYNTHESIS

1st modification in 2014, overall yield = 8% Shi, F.; Li, Z.; Kong, L.; Xie, Y.; Zhang, T.; Xu, W. Drug Discoveries Ther.
2014, 8, 117

2nd modification in 2017, overall yield = 22% Liu, F.-L.; Li, C.-Q.; Xiang, H.-Y.; Feng, S. Chem. Pap. 2017,
71, 2153
FAVIPIRAVIR’S
IMPURITIES

Diprediksi Karsinogenik

Diprediksi Mutagenik
Contoh Sintesis 6:
Qingzhong Jia, et. al., Zhongguo Yiyao Gongye Zazhi, 32(9) 385–387 (2001)
Valsartan-1

NaN3 = Sodium Azide

 Contoh Sintesis 7: Valsartan-1
• Other synthetic routes of Valsartan that avoids the use of sodium nitrite for
destroying excess NaN3 and thus prevents NDMA formation has been reported [6].
• In addition, tetrazole construction with solubilized azide source as Bu3SnN3 in
non-protic solvents (e.g.xylene and toluene) can potentially avert the formation of
NDMA (No DMF as the solvent).
• However, possible tin impurity in the final API should be considered [7,8].
• Among the various other Sartans, the possibility of NMDA generation cannot be
ruled out if NaN3/DMF conditions are used for the tetrazole construction. [9]
[6] Li, Hongwu; Yang, Hejun; Guo, Yongzheng; Jiang, Dong; Xiao, Jun; Guo, Hongju; Xu, Yongping. Method for preparing Valsartan. CN 103554049 B;
Mar 23, 2016.
[7] Padi, Pratap Reddy; Bollikonda, Satya Narayana; Jasty, Ananda Mohan; Yasareni, Suma Latha; Parmar, Vishal Dayaram. Process for preparing
Valsartan. US 7659406 B2; Feb 9, 2010.
[8] Chinta, Raveendra Reddy; Nangi, Gangadhara Bhima Shankar; Nayini, Mahendar Reddy; Yallapa, Somappa Somannavar; Budidet, Shankar Reddy;
Aminul, Islam; Meenakshisunderam, Sivakumaran. An improved process for preparation of Valsartan. US 8981109 B2; Mar 17, 2015.
[9] Vardanyan, Ruben; Hruby, Victor. Synthesis of best seller drugs. Academic Press, 2016, pp 329-356.
Contoh Sintesis 7:
Valsartan-2

Scheme 1. Synthesis of Valsartan (use of NaOCl for quenching excess

NaN3)
 Contoh Sintesis 8: Valsartan-1
• The synthesis of valsartan is discussed, inter alia, in U.S. Pat. No. 5,399,578.
• The final synthetic step (exclusive of work-up and purification) involves the reaction
of a cyano group on the biphenyl ring with an azide, for example, tributyl tinazide
(Conversion of Cyano group to Tetrazol)
Contoh Sintesis 8: Valsartan-2

Bu3SnN3 = Tributyl Tin Azide

 Persyaratan Monografi Valsartan (USP 30) dan
Impurities
• Valsartan
• L-Valine, N-(1-oxopentyl)-N-[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]-4-yl]methyl]-.
• N-[p-(o-1H-Tetrazol-5-ylphenyl)benzyl]-N-valeryl-L-valine
• CAS [137862-53-4]
• C24H29N5O3 Mr 435.52
• Valsartan contains not less than 98.0 percent and not more than 102.0
percent of C24H29N5O3,calculated on the anhydrous basis
• Related compounds—
TEST 1 (RELATED COMPOUND A): NMT 1.0%
TEST 2 (RELATED COMPOUND: B, C, OTHER RELATED COMPOUNDS
(ORC)): NMT: 0.2% A, 0.1% C, 0.1% ORC, Total NMT 0.3% (Excl. A)
•
Persyaratan Monografi Valsartan (USP 30) dan Impurities
 STUDI KASUS
1. CAPECITABINE
Sumber 1: Sumber 2:

PSD (µm): PSD (µm):

 d10 = 3,406 (n=3)  d10 = 2,474 (n=3)
 d50 = 9,917 (n=3)  d50 = 7,365 (n=3)
 d90 = 21,996 (n=3)  d90 = 19,767(n=3)
 Span = 1,874  Span = 2,348

Disolusi (Not less than 80% in 30 min. (Q)): Disolusi (Not less than 80% in 30 min. (Q)):
 98% (n=3)  98% (n=3)

Polimorfisme: Polimorfisme:
 Tidak dilaporkan ada polimorfisme  Tidak dilaporkan ada polimorfisme

Sintesis: Sintesis:
 4 Tahap (Pemula: 5-Fluorocytosine)  4 Tahap (Pemula: 5-Fluorocytosine), pada
dasarnya identik dengan sumber 1
 Rute Sintesis
Sumber 1: Sumber 2:
 Rute Sintesis
Sumber 1:
 STUDI KASUS
1. CAPECITABINE
Sumber 1: Sumber 2:

Impurities: Impurities:
 9 jenis (Sumber 1 dan 2 identik)  9 jenis (Sumber 1 dan 2 identik)

Residu pelarut: Residu pelarut:

 Metanol: NMT 3000 ppm  Metanol: NMT 3000 ppm
 Isopropil alkohol: NMT 5000 ppm  Isopropil alkohol: NMT 2000 ppm
 Metilenklorida: NMT 600  Metilenklorida: NMT 600
 Heksan: NMT 290 ppm  N-Heptan: NMT 2000 ppm
 Piridin: NMT 200 ppm  Piridin: NMT 200 ppm
 Toluen: NMT 890 ppm  Toluen: NMT 890 ppm
 Etil asetat: NMT 500 ppm  Total Residu Pelarut: 0,3%

Berdasarkan keseluruhan data, tidak terdapat perbedaan karekteristik fisikomimia CAPECITABINE

dari kedua sumber
 STUDI KASUS
2. BICALUTAMIDE
Sumber 1: Sumber 2:

PSD (µm): PSD (µm):

 d10 = 0,585 (n=2)  d10 = 0,835 (n=2)
 d50 = 1,540 (n=2)  d50 = 2,570 (n=2)
 d90 = 3,895 (n=2)  d90 = 4,730 (n=2)
 Span = 2,149  Span = 1,516

Disolusi (Not less than 80% in 30 min. (Q)): Disolusi (Not less than 80% in 30 min. (Q)):
 ND  ND

Polimorfisme: Polimorfisme:
 Bentuk I  Bentuk I

Sintesis: Sintesis:
 3 Tahap (Pemula: 3-Cyano-N-methacryloyl-3-  3 Tahap (Pemula: 3-Cyano-N-methacryloyl-3-
trifluoromethylaniline) trifluoromethylaniline) pada dasarnya identik
dengan sumber 1
 Rute Sintesis

Sumber 1: Sumber 2:
 Rute Sintesis
Sumber 1: Sumber 2:
 STUDI KASUS
2. BICALUTAMIDE
Sumber 1: Sumber 2:

Impurities: Impurities:
 6 jenis  7 jenis (6 identik dengan Sumber 1)
 Terdapat: 4-Amino-3-Trifluromethyl
Residu pelarut: benzonitrile
 Aseton
 Metilenklorida Residu pelarut:
 Diisopropil eter  Aseton
 Etil asetat  Etil asetat
 THF  Toluen
 Toluen
 Benzen (cemaran pada toluen)
Berdasarkan keseluruhan data:
 Data PSD terlalu sedikit (n=2), dari data span: terdapat perbedaan
 Informasi pada rute sintesis tidak memadai (tidak ada reagent, pelarut yang digunakan)
 Sumber 1: tidak melaporkan 4-Amino-3-Trifluromethyl benzonitrile sebagai impurities
 Perbedaan residu pelarut, data kadar/NMT tidak dilaporkan
 Tidak ada data hasil uji disolusi
Kesimpulan: perlu tambahan data