SYSTEMATIC APPROACH TO THE SAFETY ASSESSMENT OF NATURALS FOR USE IN CONSUMER PRODUCTS

SIVARAM TK

SAFETY & ENVIRONMENTAL ASSURANCE CENTRE, UNILEVER R&D, INDIA

NATURALS

Natural substances are substances which occur in nature. There are various definitions,

• According to REACH (Art. 3 (39)) [1] ‘substances which occur in nature: means anaturally occurring substance as such, unprocessed or processed only by manual,mechanical or gravitational means, by dissolution in water, by flotation, byextraction with water, by steam distillation, or by heating solely to remove water orwhich is extracted from air by any means’

• As per the European cosmetics directive, Naturals are “ Raw materials which areisolated using only physical processes from a natural source which is unchangedfrom the way it occurs in nature”.

• Naturally derived

• Natural identical

• Organic

NATURALS IN PERSONAL CARE PRODUCTS

• Naturals as cosmetic ingredients – No more a “Trend” but now in “Mainstream” with gained popularity in consumers

• There is now a growing consumer interest in food and cosmetic products which contain botanicals, often as an ingredient with established or perceived functional benefit.

• Common misconception on natural ingredients

• It’s natural so it will be safe

• It’s natural so it doesn’t contain any chemicals

• It’s easily biodegradable

• It’s a sustainable resource

ALL NATURALS - ARE SAFE !

Some see natural substances as alternatives to chemicals…

But all substances are just arrangements of the same 90-odd chemical elements.

THERE ARE NO CHEMICALS IN NATURAL PRODUCTS !

Minerals are not biodegradable!

Some essential oils not easily biodegradable

Wood does notrapidly biodegrade

NATURALS ARE EASILY / QUICKLY BIODEGRADABLE !

There can be environmental consequences…

e.g. Natural alpha-bisabolol (for sensitive skin) is extracted from Candeia plant…

…a scarce, exotic flora found in Amazonian rainforests.

ALL NATURALS - ARE EASILY SOURCED !

E.g. Natural & Synthetic fragrance & flavours

• Peppermint oil – natural: high carbon footprint – up to 50 kg CO2 equivper kg oil…

• Synthetic-derived fragrance ingredients: range from around 4 kg CO2/kg - 40 kg CO2/kg

NATURALS = LOW CARBON FOOTPRINT !

SAFETY CHALLENGES

• Naturals are complex mixtures of many chemicals, each with many potential effects, most of which are unknown

• Safety challenges are the same as those for any material• Comprehensive characterisation and risk assessment approaches• History of safe use approach

chemical complexity

chemical understanding

NATURALS

Partnering To realise full potential of naturals in a Safe and Sustainable manner

Transparency & Clear Communication on:

• Facts

• Risks

• How to manage them

Unambiguous Identification to avoid

• Adverse events

• False challenges

Specifically

• Risk & Impact Assessment Methodologies

• History of Use Data & Information

NATURALS – SAFETY – SEAC’S ROLE

Applied to Assessment of novel foods e.g. Noni Juice

Result:

Acceptable at observed intake (30 ml)*

* EU Scientific Committee for Food (2002)

History of Use

Approval decision

Evidence for

concern

Degree of similarity Exposure Biological effects

(efficacy)

Mechanism of action

Evidence of adverse

effects in Man

Toxicological data

Contra-indications

Population

No. of people exposed

Frequency of use

Intake

Duration of exposure

Origin of ingredient

Specification of

ingredient

Preparation/processing

Decision

Criteria cluster

Criterion to

be assessed

Value Tree for Safe History of Use

MCDA model

Bioavailability

… to area of acceptable HoSU

From approval decision criteria to model … Are complex and may contain

hazardous materials

Long-term human experience may be relevant measure experience

Level of safety regarded as “acceptable”

Naturals:

HISTORY OF SAFE USE MODEL

KEY CRITERIA IN DEFINING HISTORY OF SAFE USE

History of Use

Origin of ingredient

• Similarity of spec

• Prep and processing similarity

• Similarity of population to be exposed especially products aimed at babies/children (comparator should have similar history of exposure)

• No of people exposed

• Pattern of use/frequency of application

• Bioavailability/Skin penetration

Evidence of Concern

Toxicology data

• High Concern: Reproductive or developmentaltoxicity, mutagenicity, neurotoxicity or anyorgan toxicity, data showing skin sensitization(type IV allergy), type I allergy, skincarcinogenicity, phototoxicity effects

• Chemical components of concern, known skin sensitisers, photoallergens, protein

• Biological effects/mechanism of action

• Evidence of adverse effects in man(Information from literature review or existingclinical data)

R&D - SEAC

HISTORY OF SAFE USE (HOSU)

FINGERPRINTING

Knowns & unknowns – we don’t need to identify every component

• Unique signals for as many components as possible

• Holistic view

• Respectful of Ayurveda philosophy

‘A unique visual pattern representing the presence of known and/or unknown characteristic chemical components’

Definition:

ANALYTICAL DATA

R&D - SEACR&D - SEAC

COMPARATOR

PURIFIED EXTRACT

Mo

lecu

le 4

Mo

lecu

le 2

Mo

lecu

le 5

Mole

cule

7

Mo

lecu

le 8

Mo

lecu

le 2

Mo

lecu

le 4

Mo

lecu

le 3

Mole

cule

5

Mo

lecu

le 6

Mo

lecu

le 7

Mo

lecu

le 8

Mo

lecu

le 1

BUT WHAT IF

?Traditional methods Desired material

- There is no HoU for the comparator of the desired material- Or route of exposure differs compared to traditional HoU (Oral exposure but no

dermal)- For HoSU risk assessments which indicate high risk

We are good at demonstrating similarity but what if we can’t?

HOW CAN SYSTEMS BIOLOGY HELP SAFETY ASSESSMENT?

Understanding holistic and interdependent biological effects

• Identify pathways of concern

• Determine mechanism of action

• Lead further research priority areas

Naturals - When History of Safe Use is not relevant?

• Data analysis and uncertainty assessment is a challenge

We see systems biology tools and creative ways to integrate data as key tounderstanding the mechanistic effects of our ingredients and products

MULTIDISCIPLINARY RESEARCH: INVESTMENT IN PREDICTIVE BIOLOGY

• Non-animal approaches to assuring safety rely on a new network of scientific disciplines

• Exposure science• Computational/mathematical modelling • Informatics• Complex 3D cell/tissue culture/imaging• Molecular and high content biology• Transcriptomics and proteomics• Mechanistic chemistry

• New challenges around standards, quality and governance

Reynolds et al (2014), Biochemist, 36, 19-25

1. DATA GENERATION

• Start with what we know

FOCUSSED TRANSCRIPTOMICS

L1000

• L1000 Expression Profiling measures 978 validated landmark genes identified through connectivity mapping

• Comparison with public L1000 data from the LINCS Program

BioSpyder

• Custom content panels available dependant upon the problem posed

• Included in the FDA ToxCast programme

2. ANALYSIS OF MULTIPLE DATA TYPES

Microarray Differential Expression

RNAseqDifferential Expression

Whole genome methylation

Analysis

Utilise Genestack platform to manage data provenance and governance across internal & public data sets.

Functional group (pathway/Gene set) & Gene Level BMD identification for NOTEL

Differential expression

Pathway & Functional Analysis

3. INTEGRATED READ ACROSS

Lamb et al. Nature Chemical Biology 2, 663 - 664 (2006)Cronin et al. RCS Chemical Toxicity prediction: Category formation

READ-ACROSS

• Biological signatures for read-across

Readout Parameters (Biomarkers)

E-selectin

TNF-

IL-8

PGE2

IL-8MCP-1

MCP-1, IL-8, E-sel. decreaseLeukocyte recruitment

Many, e.g. Jilma et al., 2000

PGE2 decreasePain, swelling

Sebaldt et al., 1990

Collagen I & III

Collagen I, III decreaseSkin atrophy

Autio et al., 1994

MMP-1

PAI-1

SAA

PAI-1, SAA increaseCV complications

Sartori et al., 1999Fyfe et al., 1997

PAI-1

• Utilise algorithms such as Reverse Causal Reasoning to understand mechanistic causes of gene expression changes.

• Links prior knowledge of causal associations eg. Activiation of protein A results in upregulation of Gene B,C & E. Utilise directed networks.

4. MODE OF ACTION DETERMINATION AND APPLICATION

MoA or AOP

SYSTEMS BIOLOGY APPROACHES TO IDENTIFY PATHWAYS AND GAIN CONFIDENCE IN ABSENCE OF OFF-TARGET EFFECTS

Transcriptomics & metabolomics

High Content Screening

Informatics

NEXT GENERATION RISK ASSESSMENT (NGRA)

• Using new tools and approaches to build a risk assessment to enable decisions to be made

• An exposure-led risk assessment solution to biological pathway-indicated hazard concerns

Exposure led Mechanistic Hypothesis driven

PATHWAY CHARACTERISATION(TARGETS)

EXAMPLES:3D and organotypic cell modelsMolecular dynamic simulations

Integrated in vitro systems

IN SILICO-FIRST

EXAMPLES:Molecular initiating event (MIE) in

silico predictions & QSARsSkin haptenation modellingIn silico receptor screening

Tier 1

In silico-first approaches for identifying pathways of

concern, building weight of evidence and formulating

hypotheses for testing

PATHWAY IDENTIFICATION(TARGETS AND OFF-TARGETS)

EXAMPLES:HT-Transcriptomics

In vitro screening panelsReceptor binding assaysSPME free concentration

Tier 2

Identifying/characterising lead MIEs and pathways

through experimental data generation, informatics data

mining and computational modelling

Tier 3

Characterisation of response in biologically

relevant in vitro systems or complex computational

models for decision making

TIERED APPROACH

Unilever Information: Internal Use

FOR MORE INFORMATION ON SEAC’S RESEARCH VISIT WWW.TT21C.ORG