What is a nitrosamine?
FDA:
Nitrosamine impurities may increase the risk of cancer if people are exposed to them above
acceptable levels and over long periods of time, but a person taking a drug that contains
nitrosamines at, or below, the acceptable daily intake limits every day for 70 years is not
expected to have an increased risk of cancer.
• Carcinogenicity based on the fact that they can react with DNA base pairs after cytochrome P450-mediated
metabolic activation to form unstable α-hydroxymethyl-N-nitrosamines, which generate alkyl or aryl
diazonium ions as the final carcinogens.
Mutagenicity mechanism
The presence of O(6)-methylguanine in DNA
can be very harmful because methylation at
the O(6) position alters the hydrogen
bonding properties of guanine, thus inducing
G to A transition mutations. The (6) -MeG · T
is selected during replication instead of (or
preferentially) the O (6) -MeG · C pair.
Mutagenicity mechanism
EMA guideline
Sep/2019
Timeline
Valsartan
Recall
EMA, FDA
(NDMA) Jul/18
Apr/2019
ANVISA performed in loco
audits to evaluate
nitrosamines in products
which contain sartans
May/2019
Publication RDC 283
Investigation, control and
elimination of nitrosamines in
angiotensin II receptor
antagonists.
Aug/2018
Recall:
Valsartan,
Losartan e
Ibesartan
(NDEA)
Sep/2019
Recall of
ranitidine
Dec/2019
Contamination
of Metformine
(NDMA)
Ofício nº 3
Anvisa recommends
nitrosamine control by
drug manufacturers
Jan/2020
May/2019
pioglitazone
contamination
Feb/2020
Meeting between
Anvisa (GGFIS) and
regulated sector
• Official letter of entities
with evaluation proposal
and deadlineAnvisa
• Official Letter in response
to the entity Official Letter
- evaluation and
deadlines
• Mar/2020
Oct/2020
Starting ofworking
group
• Publication of the Draft for
contributions from entities
(15 days)
• extended term of RDC
283/19 - RDC 493/21
Apr/2021
Jun/2021
• Publication of Public
consultation 1050/21
Timeline
Jan/2021
Metformine call
Ofício nº 3
Anvisa recommends
nitrosamine control by
drug manufacturers
Jan/2020
• Started with EMA guidelines:
(ALL authorized human-use drugs that combine chemically synthesized APIs)
* Portfolio matrix
How Libbs deals with Nitrosamines?
2019 20202020 2021
Implementing
workflow
Step 1
• Risk assessment to identify products with a risk of N-nitrosamine formation or cross contamination.
Step 2• Confirmatory tests
Step 3• Control strategy
How to control nitrosamines?
Submitted: 27/03
12 369
Entities Request
Very high
High
Medium Low
Low
Very low
Very lowLow
Medium
Medium
Medium
High
HighVery high
Risk classification
Duration of Treatment
Max. Daily dose
Deadline for evaluation
MonthsCP 1050/2021
Dosagem Uso Crônico Pacientes tratados Risco Legenda
Baixo Não Baixo 0-Baixíssimo
Critério
Alto Médio Baixo
Baixo Sim Baixo 1- Muito Baixo Inferior Superior Inferior Superior Inferior Superior
Baixo Não Alto 1- Muito Baixo Dosagem >= 500 mg - - > 50 mg < 500 mg - - - <= 50 mg
Baixo Sim Alto 2- Baixo Utilização >= 120000 usuários - - - - - - - - - - - <= 119999 usuários
Médio Não Baixo 3- Pouco Moderado
Médio Sim Baixo 4- Moderado
Médio Não Alto 4- Moderado
Médio Sim Alto 5- Muito Moderado
Alto Não Baixo 5- Pouco Alto
Alto Sim Baixo 7- Alto
Alto Não Alto 7- Alto
Alto Sim Alto 8- Altíssimo
Entities Request
9 12 36
PRODUCTS
Months
Step 1
• Risk assessment to identify products with a risk of N-nitrosamine formation or cross contamination.
Step 2• Confirmatory tests
Step 3• Control strategy
How to control nitrosamines?
ICH M7(R1)
ASSESSMENT AND CONTROL OF DNA REACTIVE (MUTAGENIC)
IMPURITIES IN PHARMACEUTICALS TO LIMIT POTENTIAL
CARCINOGENIC RISK
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
WorkflowDMF
team
Analytical
Development
team
Packing
material
development
team
Multidisciplinary
team
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
01
02Whichnitrosamine
can be formed?
ICH M7 Classification
Retrosynthesis
Nitrosating agent
How are nitrosamines formed?
Primary, secondary or
tertiary amine and
carbonyl derivatives
Condition
Gaur, P., & Banerjee, S. (2019). Oxone-sodium nitrite mediated N-
nitrosamines formation under mild conditions from secondary amines.
Synthetic Communications, 1–10. doi:10.1080/00397911.2019.1622733
2020 Webinar: Controlling a cohort - Understanding the risk of
nitrosamines within drug substance synthesis
The common feature of these methods seems to reside in the oxidative release of the known nitrosing agents HNO2 / NO + and subsequent
formation of N-nitrosamine by reaction with secondary or tertiary amines
Other mechanisms
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
01
02Whichnitrosamine
can be formed?
ICH M7 Classification
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
01
02
Whichnitrosamine
can be formed?
ICH M7 Classification
Mutagenicidade – Teste de Ames
Teste in vitro - em bactérias
Predição in silico
Carcinogenicidade
Teste in vivo
ICH M7 (R1), 2017
Classe
1
2
3
4
5
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Hazard Identification
Based on a slide of Fernanda Waechter. Lhasa
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
Classe
1
2
3
4
5
Carcinogenic impurities
Potentially mutagenic impurities
ICH Q3A e
ICH Q3B
Nitrosamines = 18 ng/day (cronic use) ou SAR
Dose-response evaluation
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
Purge factor
Analytical results
Drying
Wash
Filtration
Centrifugation
Extraction
Reactivity
Can it be
washed?
Is it volatile?
Is it reactive?
API Impurity
Impurity level in the API or drug
2 Options
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Based on a slide of Fernanda Waechter. Lhasa
Exposure evaluation
Slide from Fernanda Waechter. Lhasa
Purge factor calculation
Initial concentration
Safe limit
Required
purge
Predicted level
Overall
predicted
purge
Initial concentration
Acceptable intake
API dose
Initial concentration
Safe limit
Purge Ratio
Overall predicted purge
Required purge
https://www.youtube.com/watch?v=OnclED3e4jc&t=299s
Exposure evaluation
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
Acceptable
Limit
Impurity level in the API or drug
Control strategy
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
* Startingmaterial
* Intermediate * API
Mutagenic impurity
• API specification control Option 1
• Intermediate specification control Option 2
• Intermediate specification control (Higher limit + purge factor)Option 3
• Purge factorOption 4
Control strategy
DS DP
Route of Synthesis
Water
IntermediateKSM
Reagents
Solvents
Materials recovery
Cross contamination Cleaning
validation
Packing material
Solvents DP Process
Cross contamination
Excipient
Excipient compatibility
Degradation
Degradation
Packing material
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
Risk Assessment
Step 1
• Risk assessment to identify products with a risk of N-nitrosamine formation or cross contamination.
Step 2• Confirmatory tests
Step 3• Control strategy
How to control nitrosamines?
• Actions for confirmatory testing (Step 2) begin immediately after identifying therisk in Step 1.
36 months to carry out Step 2 and 3 (when applicable)
Validation according to RDC 166/17 or ICH Q2
The number of batches tested must to be stablished according to risk assessment
• High sensibility of methods:
Analytical partner deadline : 90 days for results
Prospecting for more partners
Step 2- Confirmatory test
According to CP 1050/2021
Step 1
• Risk assessment to identify products with a risk of N-nitrosamine formation or cross contamination.
Step 2• Confirmatory tests
Step 3• Control strategy
How to control nitrosamines?
• 2 Scenarios:
• Presence within limits
• Presence above limits
RECALL
Carry out corrective measures
E.g.: Process changes, replacement of API, excipient or packaging material
suppliers.
- Test inclusion in Drug Product or API control
Step 3 - Control
- benefit risk assessment should be considered before recall
According to CP 1050/2021
WorkflowDMF
team
Analytical
Development
team
Packing
material
development
team
Multidisciplinary
team
Quality
Monitoring
Team
Pharmacotechnical
Team
Lessons learnt
Absence / Presence of risk
Risk Characterization
Exposure
Dose-response evaluation
Hazard Identification
To investigate the risk, first you must to
understand the ROOT CAUSE, rather than
running random tests. The root cause can be
understood only through the scientific
knowledge behind each process.
Lessons learnt
Top Related