Synoxyl AZ

®
Taking Safe Synthetics to the Next Level:
What comes after C & E?
 Genesis of Synoxyl® AZ: Molecular Design Inspired by Nature

Dehydrozingerone Synoxyl® AZ
Ginger Acetyl Zingerone

FROM…TO

Photochemically unstable O Off-white to white color solid O

Intense yellow color Photochemically stable

Prooxidant HO Not Prooxidant HO O
Phototoxic OCH3 Non-phototoxic OCH3
 Genesis of Synoxyl® AZ: Inspired by Nature

INCI name:
Acetyl Zingerone

during Purity:
>99 %
protect nurture
Appearance:
Off-white to pale yellow powder

Use level:
Before After
0.5 to 1.0%
restore

A REACH
COMPLIANT
Synoxyl® AZ’s Unique Combination of Functional Groups Neutralizes
Even the Most Reactive Forms of Reactive Oxygen Species (ROS)

ONOO - Scavenger

• H Donor Antioxidant
Chelates Fe to prevent • OH Generation via Fenton reaction Synoxyl® AZ
(•OH, •OOR)

10 Physical Quencher
2

2 Main Functions for Unique Skin Intervention

Different from
Omni-
Conventional
Antioxidant
Antoxidants
• Synoxyl® AZ operates via • Multifunctional performance of
many pathways to neutralize Synoxyl AZ differentiates it from
ROS - an Omni-Antioxidant conventional antioxidants
• Function mainly through one
pathway as an • H donor
 Synoxyl®AZ: Highly Efficient Antioxidant

Synoxyl® AZ has High Efficiency to Neutralize ROS & is

Superior to Vitamin E and Resveratrol in Chemical Assays
Unit in Peroxyl Hydroxyl Super-oxide Singlet oxygen Peroxynitrite
µmole TE/g Radical Radical Radical Non-Radical Non-Radical

Synoxyl® AZ 25,325 9,723 No effect* 7,180 1,139

α-Tocopherol 1,432 94 No effect 2,636 9

Effectiveness
17.7 39.6 NA 2.7 126.6
Ratio

Resveratrol 68.9 20.7 5.3 45.6 2.7

Effectiveness
376.6 469.7 NA 157.5 421.9
Ratio

Synoxyl® AZ provides unsurpassed capability to neutralize all of the

AZ reduced formation of superoxide
reactive forms of ROS with high efficiencies as a single ingredient
*

by inhibiting NADPH oxidase.

Distinctive Mechanisms by which Synoxyl® AZ Attenuates Reactive ROS
via Non-Sacrificial, Long-Lasting Processes

Selective Chelation of Fe Inhibits •OH Neutralizes 1O2 Mainly by Non-

Formation via Fenton Reaction Sacrificial vs Sacrificial Reactions
Fluorescence Decay Curves
Acetyl Zingerone
AZ Methyl Ether
% REMAINING DURING EXPOSURE TO 1O2
Deferiprone
EDDS
TIME (min) Acetyl Zingerone α-Tocopherol
Fluorescence intensity

Desferal
Stronger EDTA
Chelation 2 96 73
Blank

4 91 58

6 88 47

8 85 39

TIME (hours) 10 82 35

Fe+2 + H2O2 •OH + Fe+3 + -OH 1O

2 + AZ 1O
2 AZ 3O
2 + AZ

Chelation Fluorescein Decrease CHARGE - TRANSFER

Fluorescence COMPLEX
 Stressors Induce Excess ROS that Propagate Damage in Skin

External Environment

Sun Oxidative Stress Internal Environment Consequences

Pollution on the Skin
Urban
environment
Transcriptomics
Proteomics
Metabolomics

Low High
Lifestyle

Excess ROS Overwhelms Damages DNA, Proteins, Lipids • Disrupts Barrier Function
Tobacco Skin’s Antioxidant Influences Gene Expression • Drives Premature Aging
Blue light Defenses Alters Cellular Signaling • Promotes Skin Cancer
Sport
Diet
Stress

+ MORE ABOUT STRESSORS

Good News: Excess ROS Formation in Skin can be Managed
Effectively with Topical Applications
 Synoxyl® AZ Helps Skin Reduce Burden of Excess
ROS Induced by Sun Exposure in 3 Unique Ways

1 Selective Chelator

Prevents ٠OH formation via Fenton reaction Molecule

Synoxyl® AZ
Release of Free Relaxes excited states
Fe Ions back to their ground state
2 Conventional Antioxidant before they can form ROS

Fenton Molecule*
AGEs
Efficiently neutralizes ROS if formed Reaction

✕ ❸
Synoxyl® AZ
Quencher
3 Physical Quencher of Excited States ❶ ✕
Free
Synoxyl®AZ Radicals
Chelator ROS
Example: Advanced glycation endproducts (AGEs)
AGEs accumulate in abundance on collagen with aging
✕ ❷
One of the most potent sources of sun-induced ROS
Synoxyl® AZ
AZ reduces ROS from AGEs by 85% (in vitro)
Antioxidant
Skin
Damage
 Synoxyl®AZ Possesses Higher Photostability
over Conventional Antioxidants

SEE PROTOCOL
% Remaining over time Synoxyl® AZ α-Tocopherol Resveratrol

120
Inherent ~ 90% photostable
100
photostability is
80
essential for lasting
60
performance in 70% degradation in one min
40
products applied to 100% degradation in 30 mins
20
sun-exposed skin.
0
0 1 5 10 20 30
Time in minute

Aqueous ethanolic solutions were exposed to 13 J/cm2 of UV over 30 minutes

α-Tocopherol & Resveratrol degrade

completely in contrast to Synoxyl® AZ
Synoxyl® AZ Protects Keratinocytes Against Increase in Intracellular ROS
During Exposure to Blue Light or Urban Dust as Stressors

Blue light induced ROS formation 30 mins

140 60 mins
120

100
ROS level

80 In the absence of Synoxyl® AZ, each

60 stressor induced a significant
40 increase in intracellular ROS levels
20
Cells treated with Synoxyl® AZ prior
0
Blue Light
250 125 62.5 31.25 to being exposed to the stressors
- + + + + +
AZ µg/ml - - + + + + yielded significant protection - even
at the lowest dose of Synoxyl® AZ
Urban dust induced ROS formation 1 hours
40 2 hours
35 3 hours
SEE PROTOCOL
30
ROS level

25
20
15
10
5
0
250 125 62.5 31.25
Synoxyl® AZ is as Effective in Biological
Urban dust
AZ µg/ml
-
-
+
-
+
+
+
+
+
+
+
+
Systems as it is in Chemical Assays

Data represent a mean ± SD for triplicate samples.

◆◆◆◆ : p < 0.05 when compared to the untreated,
exposed group within each time interval.
Synoxyl® AZ Defends Against Formation of Cyclopyrimidine Dimers (CPDs)
During and After UV Exposure

Synoxyl® AZ reduces Keratinocytes

Synoxyl® AZ reduces AZ treated
CPDs during UV exposure Fibroblasts dark-CPD post UVA Control

exposure

0 160
-10 140 Mediated by ROS

Relative Dark-CPDs
-20 120
Relative iCPDs

100
-30
80
-40
60
-50
40
P = 0.015
-60 20
-70 0
0 1 2 4 6
-80 P = 0.002
Time post-UVA exposure in hours

Synoxyl® AZ reduces formation of iCPDs by about 70 % in keratinocytes

whereas in fibroblasts (iCPDs) & melanocytes (dCPDs) about 40%
Synoxyl® AZ Significantly Impacts Gene Expression Patterns
to Combat Hallmarks of Skin Aging

Functional Genomics
(Reconstructed Human Epidermis, Mattek EpidermFT)

Upregulates Downregulates genes associated Inhibits gene Downregulates genes

expression of “core
matrisome” genes
with ECM degradation: expression patterns associated with
metallopeptidase 3 (MMP3), associated with inflammatory responses:
cathepsin V (CTSV) and NADPH fibroblast senescence IL-17A and TGF-beta
signaling
oxidase organizer 1 (NOXO1).

Key Targets & Consequences Aging & Photo-aging Synoxyl® AZ Treatment

Core Matrisome: Collagens, proteoglycans and ECM glycoprotein — Provide youthful skin

Matrix Metalloproteases (MMPs) — Degrades ECM

Fibroblasts Senescence — Decreased collagen production & increased pigmentation

Pro-inflammatory Biomarkers — Impact on collagen production, ECM integrity & pigmentation

 Synoxyl® AZ Maintains Skin Microbiome Diversity

A Vehicle B Synoxyl® AZ

100 100
Other
90 90 Unclassified Caulobacteraceae The skin microbiome in both AZ and
80 80 Unclassified Corynebacterium vehicle control groups consisted primarily
Relative abundance

Unclassified Moraxella
70 70 of Staphylococcus spp and Cutibacterium
Unclassified Enhydrobacter
60 60
Unclassified Finegoldia
acnes and was not significantly different
50 50 Unclassified Anærococcus in composition or diversity from baseline
40 40 Unclassified Peptoniphilus to the final study visit.
Unclassitied Streptococcus
30 30
Unclassified Neisseriaceae
In fact, Synoxyl® AZ-treatment stimulated
20 20
Cutibacterium acnes skin friendly S. epidermidis by >20%
10 10 Unclassified Staphylococcus versus baseline.
0 0 Staphylococcus epidermidis
baseline 4 weeks 8 weeks baseline 4 weeks 8 weeks
Protocol

Conclusion: Synoxyl® AZ is a microbiome friendly product

Synoxyl® AZ’s Multifunctional Performance as a Skin Care Ingredient
Translate into Clinical Benefits

Manages Excess Affords Extra Protection

ROS in Skin Against DNA Damage

Synoxyl® AZ

Repairs, Maintains & Maintains

Augments ECM Integrity Microbiome Diversity
 Clinical test: Synoxyl® AZ makes Skin
More Resistant to Sun-induced Erythema

Protocol
Human volunteers Age Test sites Test substances Low UV Dose
17 35 to 55 yrs. subject's back, 0.5% Synoxyl AZ lotion 2.0 MED
4 x 2.5 cm area (No sunscreens) vs placebo

Method Results Application frequency High UV Dose

Total 9 days. On day 1 MED determined. Percent increases in ~2 mg/cm2 twice a day 2.5 MED
Then product was applied for 7 days after a* relative to baseline
which measurements were recorded with were calculated for
a Minolta Chromameter (baseline) and each product after
sites were irradiated with a low UV dose irradiation with low
(2 x MED) and a high UV dose (2.5 MED) and high UV doses.
on day 8. On day 9 Minolta Chromameter
measurements were recorded to monitor
changes in skin redness using a* values.
 Synoxyl® AZ Makes Skin More Resilient to Sun-Induced Erythema

% Reduction in a-Value Placebo

Synoxyl® AZ
(Skin Redness) 0.5% Synoxyl AZ
Does Not Absorb UV/Vis
Does Not function through a
100
39% sunscreen-type effect
90
80 Must Deliver Benefits through
its multifunctional properties
% Increase in a*

70
60
50
33%
40
30
20
10
0
Low UV Dose High UV Dose
Effect seems to be cumulative and
use of 0.5% lotion is cost-effective
p ≤ 0.05
Synoxyl® AZ: Placebo-controlled Double-blind Clinical Study Protocol

Protocol
Human volunteers Stay duration Test sites Test substances Place of Study
34 total ; Product 8 weeks plus Full face Lotion with 1% Synoxyl® AZ UC-Davis, CA
(17) & Placebo (17) prescreening & Placebo (No AZ)

Study Endpoints Assessments Application frequency ClinicalTrials.gov Identifier

• Primary Study Assessed by Board-certified Applied generously to NCT03530787
(1) Change in wrinkle dermatologist regarding signs of full face twice daily
appearance & (2) Change photoaging, including wrinkles, OFFICIAL WEBSITE ACCESS
in appearance of skin dyspigmentation, and redness with
pigmentation facial image analysis photography
and software (BTBP 3D Clarity Pro®
• Secondary Study
Facial Modeling and Analysis
(1) Change in skin redness
System (Brigh-Tex BioPhotonics)
(erythema)
Synoxyl® AZ Reverses The Key Visible Signs of Photo-aging

Vehicle
Synoxyl® AZ

P ≤ 0.05 P ≤ 0.05 P ≤ 0.005 P ≤ 0.005

Synoxyl® AZ reduces wrinkle severity and volume significantly

Synoxyl® AZ Reverses The Key Visible Signs of Photo-aging

Vehicle
Synoxyl® AZ

P ≤ 0.005 P ≤ 0.005 P ≤ 0.005

Synoxyl® AZ reduces pigment and redness intensity significantly

Synoxyl® AZ as a Multifunctional SkinCare Ingredient

Ability to All-in-one – Antioxidant, Quencher &

Selective Chelator – First-in-the Industry

Reduces Both Immediate & Dark Skin Damage

(↓CPDs) – First-in-the Industry

Photochemically Stable in Contrast to Many

Conventional Antioxidants

Maintains Integrity of Extracellular Matrix (ECM)

Clinically Validated to improve appearance of

photoaged skin, strengthen skin’s defenses
against environmental damage & maintain
diversity of skin’s microbiome
 Formulation Tips

Suggested use level

0.5 to 1.0% w/w Skin Benefits

Formulation choices Whipped Anti-Aging

Moisturizer
Daily skin protection and anti-aging applications with Synoxyl® AZ

Advices
Synoxyl® AZ is highly soluble (>20%) in quite a few
24/7 Age Defying Cream
solvents/solubilizers (see below). The material should with 1% Synoxyl® AZ – Clinically
be pre-solubilized in the emollient phase with slight Tested

heating in order to avoid crystallization in formulation

over time

Solution of Synoxyl® AZ should be added to the Broad-Spectrum

formulation, post-emulsification, preferably below 50ºC Sunscreen Oil Gel SPF 30+
with Synoxyl® AZ and Synoxyl® HSS
Finished product must be acidic:
pH<6.5 is recommended

SEE SKIN BENEFITS

SEE SOLUBILITY PROFILE
 Publications

Acetyl zingerone: An efficacious multifunctional ingredient for continued

protection against ongoing DNA damage in melanocytes after sun exposure ends + SEE PUBLICATION
R.K. Chaudhuri*, T. Meyer*, S. Premi† and D. Brash‡
International Journal of Cosmetic Science, 42(1):36-45, 2020

Omni Antioxidant Acetyl Zingerone Scavenges /

Quenches Reactive Species, Selectively Chelates Iron + SEE PUBLICATION
Thomas Meyer and Ratan K. Chaudhuri
Cosmetics & Toiletries Vol. 135, No. 9 | October 2020

Randomized Double-Blind Vehicle Controlled Study

of The Effects of Topical Acetyl Zingerone on Photoaging
Simran Dhaliwal BS, Iryna Rybak BS, Aunna Pourang MD, Waqas Burney MBBS, Kelly Haas PhD, + SEE PUBLICATION
Simran Sandhu BS, Robert Crawford PhD, Raja K Sivamani MD, MS, AP
Journal of Cosmetic Dermatology doi:10.1111/JOCD.13464

A Zingerone Analog, Acetyl Zingerone, Bolsters Matrisome Synthesis, Inhibits

Matrix Metallopeptidases, and Represses IL-17A Target Gene Expression + SEE PUBLICATION
William R. Swindell, Krzysztof Bojanowski and Ratan K. Chaudhuri
Journal of Investigative Dermatology, 140:602-612, 2020
 Questions?

Tom Meyer, PhD

Technical Adviser
info@sytheonltd.com

Sytheon Ltd., USA Sytheon SARL, France

info@sytheonltd.com sytheon@sytheonltd.com
973-988-1075 +33 1 41 10 81 82

www.sytheonltd.com
 Main Environmental Stressors That Induce ROS Formation in Skin

External Environment

Sun is the most potent generator of ROS

Sun Oxidative Stress Internal Environment Consequences
Pollution • At least 50% of sun-induced ROS on the Skin
Urban originates from visible radiation
environment
• Outside the protective capacity Transcriptomics
of sunscreens Proteomics
Metabolomics
Estimated that Sun accounts for 80% of
Low Highof aging
visible signs
Lifestyle
Particulate matter linked with
Excess of Reactive
hyperpigmentation & wrinklingDamages DNA, Proteins, Lipids • Disrupts Barrier Function
Tobacco Oxygen Species (ROS) Influences Gene Expression • Drives Premature Aging
Blue light Altersfactors
Cellular Signaling • Promotes Skin Cancer
Sun & pollution are cited as major in
Sport
Diet the aging skin exposome
Stress

+ MORE ABOUT STRESSORS

 Synoxyl®AZ Possesses Higher Photostability
over Conventional Antioxidants

SEE PROTOCOL
% Remaining over time
Protocol Synoxyl® AZ α-Tocopherol Resveratrol

All
120samples were prepared in 50 % ethanol in 5mL OPTICLEAR© vials
Inherent (KIMBLE glass, Chicago Heights, IL), test vials were then placed in a ~ 90% photostable
100
Rayonet RPR-100 photochemical reactor equipped with four RMR-3000
photostability is (UVB) and four RMR 3500 (UVA) lamps (Southern New England
80
essential for lasting Ultraviolet Company, Branford, CT), the combination of UVA and UVB
simulated
60 day light condition. 100 μg/mL of the sample was irradiated
performance in at 31 °C at a dose70% degradation in one min
of 13 J/cm2.
40
products applied to Synoxyl ® AZ was Trans-Resveratrol α-tocopherol was 100% degradation in 30 mins
20
sun-exposed skin. quantified by HPLC was quantified by quantified by HPLC
according
0 to “Method HPLC according to at 295nm using
of Analysis of0 Acetyl internal
1 method5 100%10methanol as 20 30
Zingerone content in SOP #902-01. the mobile phase.
Time in minute
Synoxyl AZ.”

Aqueous ethanolic solutions were exposed to 13 J/cm2 of UV over 30 minutes

α-Tocopherol & Resveratrol degrade

completely in contrast to Synoxyl® AZ
Synoxyl® AZ Protects Keratinocytes Against Increase in Intracellular ROS
During Exposure to Blue Light or Urban Dust as Stressors

Blue light induced ROS formation Media 30 mins

60 mins
140
EpiLife Media (60 µM Calcium) supplemented
120
with 0.2% bovine pituitary extract
100
ROS level

80 In the absence of Synoxyl® AZ, each

60 Cells stressor induced a significant
40 Epidermal Keratinocytes loaded with DCF increase in intracellular ROS levels
20 (2’, 7’-dichloro-dihydroflurescein diacetate DCF)
Cells treated with Synoxyl® AZ prior
0
250 125 62.5 31.25 to being exposed to the stressors
Source of Blue Light
Blue Light - + + + + +
AZ µg/ml - - + + + + yielded significant protection - even
LED Panel composed of 226 blue LED lights at the lowest dose of Synoxyl® AZ
Urban dust induced ROS formation arranged in a square
1 hours 15x15 LED matrix with an
40 2 hours
output of 22 wats of 450 nm blue light
35 3 hours
SEE PROTOCOL
30

Source of Urban Dust

ROS level

25

Sigma (standard reference1649b);

20
15
10
prepared at 10 mg/ml in the media
5
0
250 125 62.5by BioInnovation
31.25
Synoxyl® AZ is as Effective in Biological
Work done Laboratories, Inc.
Urban dust
AZ µg/ml
-
-
+
-
+
+
+
+
+
+
+
+
Systems as it is in Chemical Assays

Data represent a mean ± SD for triplicate samples.

◆◆◆◆ : p < 0.05 when compared to the untreated,
exposed group within each time interval.
 Synoxyl® AZ Maintains Skin Microbiome Diversity

A Vehicle B Synoxyl® AZ

100 100 Nasolabial and glabellar skin were swabbed under

Other
90 90 sterile conditions with sterile swabs
Unclassified to collect
Caulobacteraceae The skin microbiome in both AZ and
Unclassified Corynebacterium vehicle control groups consisted primarily
80 80 specimens for microbiome analysis at baseline, visit
Relative abundance

Unclassified Moraxella
70 70
2, and visit 3. Swabs were stored at -80°C until DNA of Staphylococcus spp and Cutibacterium
Unclassified Enhydrobacter
60 60
was extracted. Unclassified Finegoldia
acnes and was not significantly different
50 50 Unclassified Anærococcus in composition or diversity from baseline
40 40 In order to assess for unintended shifts of the
Unclassified Peptoniphilus to the final study visit.
30 30 microbiome, the skin microbiome was analyzed at
Unclassitied Streptococcus
In fact, Synoxyl® AZ-treatment stimulated
Unclassified Neisseriaceae
20 20 baseline and at follow up. Each sample resulted in an skin friendly S. epidermidis by >20%
Cutibacterium acnes
10 10 average of 15,845 reads (5,456 Staphylococcus
Unclassified to 24,064). versus baseline.
0 0 Staphylococcus epidermidis
baseline 4 weeks 8 weeks baseline 4 weeks 8 weeks
Protocol

Conclusion: Synoxyl® AZ is a microbiome friendly product

 Formulation Tips

Suggested use level

0.5 to 1.0% w/w
Synoxyl® AZ Solubility Profile
Skin Benefits
SOLUBILIZERS SOLUBILITY OF AZ
Formulation choices Dimethyl Isosorbide
Whipped Anti-Aging
Moisturizer 25%
Daily skin protection and anti-aging applications with Synoxyl® AZ

Ethoxydiglycol 25%

Advices Dipropylene Glycol 20%

Synoxyl® AZ is highly soluble (>20%) in quite a few
24/7 Age Defying Cream
solvents/solubilizers (see below). The material should
Ethanol with 1% Synoxyl® AZ – Clinically 20%
be pre-solubilized in the emollient phase with slight Tested
Propylene Carbonate
heating in order to avoid crystallization in formulation 20%
over time
Dipropylene Glycol Dibenzoate and Di-Propylene Glycol
20%
Solution of AZ should be added to(2:1)
Synoxyl® the Broad-Spectrum
formulation, post-emulsification, preferably Dipropylene
below 50ºC Glycol Dibenzoate Sunscreen Oil Gel SPF 30+ 15%
® ®
with Synoxyl AZ and Synoxyl HSS
Finished product must be acidic:
Isosorbide Dicaprylate (HydraSynolTM DOI) 7%
pH<6.5 is recommended

SEE SKIN BENEFITS

SEE SOLUBILITY PROFILE
 Formulation Tips

Suggested use level

0.5 to 1.0% w/w Skin Benefits

Moisturizer
Formulation choices Restorative
Whipped Anti-Aging Sunscreen
Moisturizer
Daily skin protection and anti-aging applications with Synoxyl® AZ

Couple enhanced Actively repair existing signs Extend protection

Advices hydration with daily of photodamage or aging for capacity of sunscreens
Synoxylproactive skin care.
® AZ is highly soluble (>20%) in quite a few
a younger-looking skin. from sun damage.
24/7 Age Defying Cream
solvents/solubilizers (see below). The material should with 1% Synoxyl® AZ – Clinically
be pre-solubilized in the emollient phase with slight Tested

heating in Strengthen defenses

order to avoid crystallization in formulation Reduce appearance of wrinkles Help skin cope with excesses
over time Nurture ECM - stop degradation of sun-induced ROS
Even skin tone
Solution ofMaintain
Synoxyl®younger-looking
AZ should be added
skin to the Prevent dark DNA damage
Diminish rednessBroad-Spectrum after sun exposure stops
longer over the course of life
formulation, post-emulsification, preferably below 50ºC Sunscreen Oil Gel SPF 30+
with Synoxyl® AZ and Synoxyl® HSS
Finished product must be acidic:
pH<6.5 is recommended

SEE SKIN BENEFITS

SEE SOLUBILITY PROFILE