IceAwake™

Fighting aging
in sleep-deprived skin
IceAwake™
Fighting aging in sleep-deprived skin

Extract from a Glacier Bacterium to IceAwake™

Wake up Sleep-Deprived Skin Cells
IceAwake™ is based on an extract of the bacteria
­Iodobacter ssp., which was isolated from the soil
below a Swiss glacier. IceAwake™ rejuvenates skin
that appears aged due to a hectic lifestyle that
also features too little sleep.
Sleep deprivation not only leads to a tired appearance
in the short-term, but it is also an aging factor in the
long-term. This is due to inefficient protein folding in
sleep-deprived and aged skin. Unfolded proteins are
­unable to carry out their job and accumulate. This leads
to stress in the endoplasmic reticulum (ER), the cellular
organelle responsible for secreted protein production,
and subsequently to cell damage. In addition, ATP,
the energy molecule that is needed for protein folding
and many other cellular processes, is reduced in sleep-
deprived cells.
IceAwake™ was shown to:
• increase the expression of chaperones in aged
­fibroblasts that assist in proper protein folding
• increase ATP production and reduce ER stress
in a sleep deprivation cell model.
In clinical studies on Asian as well as Caucasian female
and male volunteers, IceAwake™ visibly rejuvenated
their appearance by decreasing wrinkle depth, increas-
ing radiance, and reducing the tired appearance for
a refreshed look.
• Energizes tired skin
• Reduces wrinkles after only two weeks
• Increases radiance despite a hectic lifestyle
Applications
• Energizing serums for fatigued skin
• Concentrates for glowing, revitalized skin
• Radiance enhancing face masks
• Anti-aging treatments
Formulating with IceAwake™
• Recommended use level: 1 – 2 %
• Incorporation: For cold processes, dissolve ­IceAwake™
into the aqueous phase or add it p ­ re-dissolved into
a small amount of water after e ­ mulsification. For hot /
cold processes, add I­ceAwake™ pre-dissolved in
a small amount of water, during the cooling phase
­below 40 °C.
• Thermostability: Temperatures of up to 40 °C for a
short time will not affect the stability of IceAwake™.
INCI (EU / PCPC) Declaration
Succinic Acid (and) Maltodextrin (and) Aqua / Water
Additional Information
• Without preservation
• Without alcohol

IceAwake™ is COSMOS approved *.

* Raw material approved by ECOCERT GREENLIFE in accordance with the

COSMOS Standard December 2019
IceAwake™
Swiss glacier bacteria reawakened
A Glacier Expedition to Discover new Microorganisms
In order to discover and harvest novel extremophile
­microorganisms for use in cosmetics, an expedition was
undertaken to a glacier in Valais, Switzerland. Due to the
constant shrinking of glaciers in the past few decades,
microbes that were previously hidden below a perma-
nent layer of ice have now become accessible. Samples
taken from the soil were analyzed for their microbial
­content. Whole genome sequencing of an isolated strain
led to the identification of Iodobacter ssp., a psychrotol-
erant rod shaped gram-negative bacterium, which has
been reawakened following many years lying dormant
below the glacier ice layer.
Endoplasmic Reticulum (ER)
IceAwake™ Rejuvenates Sleep-Deprived Skin
Large scale cultivation and extraction of the
Iodobacter ssp. strain followed by spray-drying
the extract on a maltodextrin carrier yields the
active ingredient IceAwake™.
IceAwake™ contributes to the improvement of
correct protein folding, reduces stress, and
stimulates energy production in vitro in sleep-
deprived cell models.
In two clinical studies with Asian and Caucasian
female and male volunteers, IceAwake™ reju-
venated their ­appearance after just 14 days by
reducing wrinkle depth and increasing radiance
for a visibly less tired look.
Membrane
Cytoplasm
Mitochondrion
Nucleus
Endoplasmic reticulum (ER)
Sleep Deprivation
A hectic lifestyle leads to skin aging
Lack of Sleep is Part of our Lives
In our modern society, many people do not achieve
the recommended level of 7 – 9 hours of sleep per night.
A hectic lifestyle, stress and poor sleep hygiene prevent
both the quality and quantity of sleep that the body
needs. The skin is considerably affected by this sleep
deprivation and the most obvious effect is the tired
­appearance of the face following a night featuring low
sleep quantity and / or quality. Whilst young individuals
can recover quickly from this lack of adequate rest, it
is important to note that prolonged sleep deprivation
acts as a general aging factor like UV-irradiation or
­oxidative stress, which leads to long-term negative
­effects on the skin.
Protein Folding in the ER
The endoplasmic reticulum (ER) is an organelle in our
cells which, among other important functions, produces
proteins that are destined for cellular membranes and for
secretion, e. g. collagen. During the production process,
these proteins need to be correctly folded in order to
perform their intended function. There are helper pro-
teins known as chaperones and they assist in this folding
process. BiP (binding immunoglobulin protein) is one of
the major chaperones in the ER and it has been shown
to be specifically affected by sleep deprivation (1). It has
also recently been shown that BiP is, for example, up-
regulated before the increased production of collagen
at night during cellular regeneration processes (2).
Protein Folding in the Endoplasmic Reticulum
Chaperones
Protein folding
Unfolded protein
Hectic lifestyle Protein
Stress misfolding
Aging
Sleep Deprivation and Protein Folding
Sleep deprivation affects protein folding and chaperone
production, and it leads to stress in the ER. In order to
combat this issue, more chaperones are needed to cope
with the stress and assist in the regeneration and repair
of the cell. In aged cells, chaperone production is reduced
and the cell fails to efficiently prevent protein misfolding.
These wrongly folded proteins accumulate and cause
more ER stress, and subsequently damage the cell. This
vicious cycle then leads to an insufficient recovery of the
cells during sleep.
Sleep Deprivation leads to Cellular Burnout
An additional negative consequence of sleep depriva-
tion is the reduction of the cellular energy currency
ATP produced by mitochondria (3). As the chaperones
in the ER are ATP dependent, insufficient ATP levels
due to lack of sleep can limit their protein folding ability
and exacerbate the accumulation of misfolded proteins
which ultimately leads to cell damage.
Improving the flawless folding of proteins and increasing
ATP levels results in a rejuvenated and fresh appearance
despite a hectic lifestyle.
(1) Naidoo et al., J Neurosci. 2008; 28(26): 6539 – 6548
(2) Pickard et al., FASEB J. 2019;33(6): 7479 – 7489
(3) Chikahisa and Séi, Front Neurol. 2011; 2: 87
ATP (energy)
Active protein ĺ radiant, smooth skin
ĺ youthful appearance
Accumulation ĺ tired looking skin
Degradation ĺ wrinkles
ER stress ĺ aging
Cell damage
IceAwake™
Study results

Improving Chaperone Expression

in Aged Fibroblasts
As it is known that ER chaperones are down- Fibroblasts treated with 1 % Iodobacter ssp. extract
regulated in aged cells, which consequently results in showed an increase in the expression of all four
an accumulation of misfolded proteins, we wanted to ­chaperon proteins compared to control cells.
­investigate whether an extract of Iodobacter ssp. could
increase the expression of chaperone genes in aged BiP, in particular, which is the main chaperone that is
skin cells. For this study, fibroblasts were grown for ­responsible for the unfolded protein response in the
17 passages to mimic the aging process. These aged ER, was upregulated by 100 %. This indicates that
­fibroblasts were then treated or not (control) with 1 % ­protein folding can be carried out more efficiently
­Iodobacter ssp. extract for a period of 24 hours. The ­despite aging.
­expression of four ER chaperone genes was quantified
with RT-qPCR.

BiP is the main chaperone in the ER. It not only assists

in protein folding, but it also keeps proteins from
­aggregating and imports polypeptides into the ER.
Endoplasmin plays a critical role in folding proteins that
are secreted.
Calnexin and calreticulin are both important for protein
quality control as they prevent the release of misfolded
proteins from the ER.

Improving Chaperone Expression in Aged Fibroblasts

Control 1 % Iodobacter ssp. extract

250
compared to control cells (=100) in %

200
Gene expression

150

100

50

0
Control BiP Endo- Calnexin Calreticulin
plasmin

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 Increase in ATP Production in a Cellular Sleep
Deprivation Model
A cellular sleep deprivation model was used Treatment of sleep-deprived fibroblasts with 0.01 %
in order to mimic the effects of lack of sleep in a cell Iodobacter ssp. extract significantly stimulated the
­culture environment. These special “sleep-deprived” ­production of ATP by 27.5 %.
­fibroblasts combine the impairment of mitochondria and
increased ER stress which mimics the sleep deprivation
phenotype. Using this cellular model, it was investigated
whether the Iodobacter ssp. extract is able to reactivate
ATP production.

The sleep-deprived fibroblasts were grown in assay

­medium that ­induces additional mitochondrial stress.
­After washing, cells were incubated or not (control)
with 0.01 % Iodobacter ssp. extract for 120 minutes.
­Cellular ATP levels were measured by chemilumines-
cence.

Increase in ATP Production

Control 0.01 % Iodobacter ssp. extract

150

* + 27.5 %
compared to control cells (=100) in %

100
ATP production

50

*p<0.05 versus control

S-809 / © Mibelle Biochemistry

IceAwake™
Study results

Reduction of ER Stress
The same cellular sleep deprivation model as As expected, the sleep-deprived fibroblasts displayed
in the previous study was used in order to a larger n
­ umber of MAMs compared to healthy fibro-
mimic the effects of lack of sleep in a cell culture envi- blasts. The treatment of sleep-deprived fibroblasts with
ronment. These special “sleep-deprived” fibroblasts 0.01 % Iodobacter ssp. extract significantly reduced
combine the impairment of mitochondria and increased MAM contact points.
ER stress which are consequences of a chronic lack of
sleep. This suggests that ER stress in the sleep-deprived cell
model has been decreased by the Iodobacter ssp. ­extract.
Using the model, the formation of mitochondrial asso­
ciated membrane (MAM) contact points were investi-
gated. These MAMs are the connection points between
mitochondria and the ER, and they act as a marker for
ER stress. As these contact sites contribute, among
other things, to the detoxification of accumulated mis-
folded proteins, MAMs are increased under ER stress.

In this study, sleep-deprived fibroblasts were treated

or not (control) with 0.01 % Iodobacter ssp. extract for
120 minutes and MAM contact points were visualized
and quantified using a proximity ligation assay followed
by immunofluorescence microscopy. The number of
­cellular MAMs was compared to normal fibroblasts which
do not have elevated ER stress.

Reduction of Mitochondrial Associated Membrane (MAM) Contact Points in Sleep-Deprived Fibroblasts

MAM contact points Cell nuclei

250 Control healthy

compared to healthy cells (=100) in %

200
Change in protein interactions

150
10 μm

100
Sleep-deprived cells
0.01 %
50 Control Iodobacter ssp. extract

0
Control healthy Control 0.01 % Iodobacter
ssp. extract
Sleep-deprived cells

*p<0.05 versus control sleep-deprived cells

S-809 / © Mibelle Biochemistry S-809 / © Mibelle Biochemistry
 Improvement of Wrinkle Depth and Radiance
in Asian Volunteers with a Hectic Lifestyle
Twenty-three Asian women aged between These results demonstrate that after just 14 days of
41 and 57 years (mean age: 50.7 years) who were over- ­application, IceAwake™ rejuvenates the appearance
worked and/or had little and bad quality sleep were of the face despite a hectic lifestyle by decreasing
­selected for the study. They applied a cream with 2 % ­wrinkles and improving skin radiance.
IceAwake™ on one half of their face and the corre-
sponding placebo on the other half of their face twice
daily for a period of 14 days. The wrinkle depth on
the crow’s feet wrinkles was determined by using
­PRIMOS. In addition, a clinical expert grading of skin
radiance was performed based on photographs taken
of the volunteers using Visia CR.

Treatment with 2 % IceAwake™ for 14 days increased

skin radiance by 9.2 % and reduced wrinkle depth
by 11.4 %. Both results were statistically significant
­compared to initial conditions and placebo.

Decrease in Wrinkle Depth after 14 Days Increase in Radiance after 14 Days

Placebo 2 % IceAwake™ Placebo 2 % IceAwake™

5 10
*
Change in maximum wrinkle depth
compared to initial conditions in %

compared to initial conditions in %

8
Improvement of skin radiance

-5

-10
2
*

-15 0

*p<0.05 versus initial conditions and placebo *p<0.05 versus initial conditions and placebo

S-808 / © Mibelle Biochemistry S-808 / © Mibelle Biochemistry

IceAwake™
Study results

Improvement of Wrinkles and Tired

­Appearance in Female and Male Volunteers
Twenty-one Caucasian men and women aged After 14 days of treatment with 2 % IceAwake™, a signif-
between 44 and 66 years (mean age: 53.7 years) who icant reduction of wrinkle depth by 15.9 % compared to
had a tired appearance (presenting dark circles around initial conditions was observed.
the eyes and medium-deep crow’s feet wrinkles) were
selected for the study. They applied a cream with 2 % In addition, treatment with 2 % IceAwake™ significantly
IceAwake™ on one half of their face and the corre- reduced visible tiredness in 71 % of the volunteers as
sponding placebo on the other half of their face twice ­assessed by clinical-grade evaluation.
daily for a period of 14 days. The wrinkle depth on the
crow’s feet wrinkles was determined by using PRIMOS.
In addition, a clinical expert grading of skin tiredness
was performed on photographs taken of the volunteers
using ColorFace.

Decrease in Wrinkle Depth after 14 Days Decrease in Tired Appearance after 14 Days

Placebo 2 % IceAwake™ Placebo 2 % IceAwake™

10 100
Change in wrinkle depth compared to initial

Volunteers showing visible improvements

75
*
0
of tiredness in %
conditions in %

50

-10
25

-20 0

*p<0.05 versus initial conditions *p<0.05 versus initial conditions

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Furthermore, the wrinkle and tiredness reduction The perception of each volunteer regarding efficacy
was visible in the ColorFace photographs of both was also evaluated via a questionnaire at the end
male and female volunteers. of the study.

95 % of the volunteers agreed with the following

­statements:
“The product helps my skin to look healthy”
“The product makes my skin feel soft and smooth”
“The regular use of the product has had an overall
­benefit on my skin”

90 % of the volunteers agreed with the following

­statements:
“The product provides a healthy radiance (‘glow’)”
“The product instantly provides me a skin that looks
both healthy and beautiful”
“The product helps to make my skin feel fit for the day”

Visible Improvement of Crow’s Feet Wrinkles in Male and Female Volunteers

Before After 14 days

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 IceAwake™
Fighting aging
Mibelle AG Biochemistry, 5033 Buchs  /  Switzerland, Phone + 41 62 836 17 31, info@mibellebiochemistry.com, www.mibellebiochemistry.com

in sleep-deprived skin

IceAwake™
• Energizes tired skin
• Reduces wrinkles after only two weeks
• Increases radiance despite a hectic lifestyle

Applications
• Energizing serums for fatigued skin
• Concentrates for glowing, revitalized skin
• Radiance enhancing face masks
• Anti-aging treatments

Marketing benefits
• Based on a unique extremophile bacterium found under a Swiss glacier
• Sustainable sourcing of raw material (biotechnology)
• Novel aging mechanism based on protein folding and chaperone activity
• Without preservatives
• COSMOS approved

Innovating for your success

Mibelle Biochemistry designs and develops innovative, high-quality actives based on
naturally derived compounds and profound scientific know-how. Inspired by nature –
Realized by science.

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erty rights. Mibelle AG Biochemistry will not assume any expressed or implied liability in connection with any use
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