CM-Glucan

Sodium Carboxymethyl Betaglucan

 CM-Glucan

New potent biological agent

Cosmetic and dermatological applications:
- Anti-aging
- Sun Care
- Baby Products
- Wound healing
- Post laser treatments
 CM-Glucan

Meets the demands of today's

consumers for active natural
compounds
Acts in a holistic way by inducing the
body's own defense mechanisms
Is prepared from renewable resources
 CM-Glucan
Sodium Carboxymethyl Betaglucan

CH2OCH2COO Na
O CH2OCH2COO Na
O
O O
OH
O
OH OH
OH

CH2
CH2OCH2COO Na

O O

O O
OH CH2OCH2COO Na

OH O O
OH
OH O
OH
OH

Polysaccharide from baker's yeast (Saccharomyces cerevisiae)

β-(1-3)-linked polyglucose
Carboxymethylation (water soluble derivative)
 Glucan

1941 first defined pharmaceutical yeast product

(Zymosan)
Identification of glucan as the immunologicaly
active part of Zymosan
Glucan belongs to the class of drugs known as
biological response modifiers (BRMs)
BRMs stimulate the body own defense mechanism
Glucan preparations enhance macrophage-
mediated phagocytosis and the production of
cytolytic factors
 Cytolytic Cytostatic Factors
glucan
receptor
GLUCAN inflammatory
response
macrophage

leukotrienes

enhanced
activated macrophage phagocytosis

white blood cell

proliferation
T-cell stimulation
interleukin 1 colony stimulating
factors
Beta-1,3-Glucan in Research

Indication:
Acceleration of wound healing
Higher resistance to bacterial infections
Drug delivery system

Publications (last ten years):

more than 40 patents
more than 1000 papers
 Activity of CM-Glucan

Increase of life-span of L-1210 leukemia bearing cells in mice at a

dose of 1450 mg/kg
Kery V. et al., Int. J. Biochem., 22, 1203-1207, 1990

CM-Glucan potentiates an antibacterial effect of antibiotics and has

radioprotective effects
Horvathova M., et al., Carbohydrate Polymers, 15, 79-87, 1991

Cyclophosphamide treated mice exhibited less pronounced

immunosupression and more rapid hematopoietic recovery when
administered with CM-Glucan
Wagnerova J., et al., Immunopharm. and Immunotox., 15, 227-242, 1993
Wound Healing Properties of Glucan Preparations

Significant acceleration of wound healing could be detected in the animal leg

model by topical application of yeast glucan preparations at different
concentrations (20-100 ug/ml)
Wolk, M. and Danon, D. Med. Biol. 63, p 73-80 (1985)

Glucan exhibits a particularly marked enhancement of the proliverative

phase of wound healing in the mice skin model at a concentration of 7.5 mg/ml
Leibovich, S. J. and Danon, D. J. Reticuloendothel. Soc. 27, p 1-11(1980)

Topical application of glucan preparations enhance wound breaking strength

in rats by 270% due to an increased macrophage function.
Browder, W., Williams, D., Lucore, P., Pretus, H., Jones, E. and McNamee, R. Surgery
104, p 224-230(1988)
Preparation of CM-Glucan

Isolation of Glucan from

the cell wall of baker's yeast

Derivation to carboxymethyl glucan

Degree of substitution 0.75
Characterization of the polymer structure
by 13C-NMR
Determination of the active structure
 Active Structure of CM-Glucan

Helix-Coil Transition
Wavelength (nm)
515
510
505
500
495
490
485
480
475
470
0.001

0.01

0.1

Concentration

1.0
NaOH (M)
CM-Glucan Laminarin
Dextran Congo Red
 Solar Radiation

sunlight

UVC UVB UVA visible light infrared

290 320 400 750 wavelength (nm)

UV-C Screened out by the atmosphere

UV-B Melanization
Sunburn
Premature skin aging
Immuno-Suppression
Skin tumors
 Solar Radiation

sunlight

UVC UVB UVA visible light infrared

290 320 400 750 wavelength (nm)

UV-A Oxidative Stress, Photo Aging

Photo Toxicity, Photo Allergy

Visible and Unknown, not investigated

infrared
Immuno-Suppression

UV-B radiation indues a suppression of the

immunological activity within the epidermis

The viability of the immunocompetent cells

(keratinocytes and Langerhans cells) is
impaired

Immuno-suppression supports the

proliferation of skin cancer
 Immuno-Suppression

Sunscreens do not offer protection against all

biological effects of sunlight

Based on this fact the use of sunscreen products

with high SPF values might be dangerous

The use of stimulators of the immune system in

sun care formulations is a new approach to lend
beneficial functions to these products
Protection of Human Cultured Cells Against Oxidative Stress
Including Ultraviolet Radiation

Cell Cultures
Fibroblasts and keratinocytes
of normal adult human skin
UV-Radiation
Samples
UV-A (320 nm - 450 nm)
Glucan dispersions and CM-
300 W/m2
Glucan solutions at a final
concentration of 100 μg/ml
(corresponding to a 0.5% End Points
application of the product CM- Intracellular concentrations of
Glucan (2% solution) Glutathione (GSH) Ferritin
Influence of CM-Glucan regarding glutathione
in human keratinocytes

400
GSH (nmol/mg protein)

300
Non treatment
Alpha-tocopherol
CM-Glucan
200

100
0 2 4 6 8 10
5 2
UVA Fluence 10 J/m
320 - 450 nm
Influence of CM-Glucan regarding ferritin
in human fibroblasts

500

Non treatment
400
Ferritin (ng/mg protein)

Alpha-tocopherol
CM-Glucan
300

200

100
0 2 4 6 8 10
5 2
UVA Fluence 10 J/m
320-450 nm
Photoprotective Effects

CM-Glucan in combination with UV-A radiation

does not cause any additional photo toxicity to
keratinocytes and fibroblasts

CM-Glucan clearly has protective effects regarding

oxidative stress induced by UV-A radiation

CM-Glucan is active at concentrations as low as

100 ug / ml (0.01%)
Inhibition of lipid peroxidation

Squalene peroxides [units CL] 7

6
5
4
3
2
1
0
nt nt 0% 0,04% 0,2%
CM-Glucan
in o/w emulsions
2
nt UV-A 10 J/cm
Inhibition of squalene peroxidation by formula containing
different antioxidants or CM-Glucan

‹ Caffeine
% inhibition of squalene peroxidation

100
our study ‹ Tetramethylpiperidine
‹ Tocopherol Acetate
80 ‹ t Butylphenylnitrone
Colin et al. ‹ Butylhydroxytoluene
‹ 4-hydroxy TPO
60
‹ Propylgallate
‹ Doxy cyclohexane
40 ‹ Ascorbic acid
‹ DLa-tocopherol
20
‹ b-carotene
‹ butulated hydroxy anisole
‹ Da-tocopherol
0 ‹ mixture of tocopherols
TPO

HTPO

DHNO
TBP
BHT

0.2% CMG
Car
Vit C

Toc
Gallate

Vit En

0.04% CMG
BHA
Vit E Ac

Vit Es
CAFF

‹ 0.04% CM-Glucan
‹ 0.2% CM-Glucan
antioxidants at 0.2%
 Oxidative Stress

UVA UVB

UV-A photons are present in sunlight in

much higher quantities than those of
UV-B and penetrate deep into the
dermis.

UV-A generates active oxygen

molecules (singlet oxygen, hydroxyl
radicals).
 Oxidative Stress

Active oxygen molecules damage skin cells

and cause lipid peroxidation, immuno-
suppression, loss of skin elasticity and even
carcinogenesis.
Long-term UV-A irradiation is responsible for
skin aging.
 Oxidative Stress

CM-Glucan activates the skin’s self defense

mechanisms. The endogenous antioxidant
levels (e.g. glutathione) are enhanced and
secondary stress markers (e.g. ferritin) are
induced.
CM-Glucan's profile in cosmetic formulations

Immune stimulatory activities

Sun care products, daily creams

Wound healing properties

After sun, after shave, impure skin treatment

Treatment
prophylactic and therapeutic activity
daily use
long-term activity
 Possible role of keratinocytes on
the skin's immune system

Endotoxins
UV
Chemicals

KC
KC
ICAM-1
Expression
IL-1 Macrophages
EC
TNF- a Activation of
IFN- g
T-Cells

Recruitment of

Leucocytes
 Oxidative Stress

UV-A Irradiation

CMG
CMG CMG CMG Stratum
CMG
CMG CMG Corneum
Keratinocytes CMG CMG
Glutathione
Vitamine E
Epidermis
Langerhans Cells CMG

Heme Oxygenase Cytokines

Dermis
Fibroblasts Ferritin
Placebo controlled Studies with CM-Glucan
on Human Skin

Concentration range:
0.04 - 0.4% CM-Glucan
Formulation as hydrogel or o/w emulsion
Application twice daily
Control panel five to ten healthy volunteers
Enhanced Skin Proliferation
40
(%) Increase of the cell renewal rate µ

µ = ln 2 / tr
30

20

10

0
0% 0.1% 0.4% 0.04% 0%
hydrogel o/w emulsion

CM-Glucan
 Growth of Keratinocytes
with CM-Glucan

150
relative cell count in %

control
20 mg/L
125
6.6 mg/L
5 mg/L
4 mg/L

100

75
24 48 72 96 120
time (h)
 Skin firmness

100
Increase in skin firmness [%]

0.04% CM-Glucan
80
Placebo
60 untreated

40

20

0
14 days 28 days
-20
Reduction of wrinkles

[%]
20
0.04% CM-Glucan
15 Placebo

10

-5
14 days 28 days
 Post laser treatment

Improvement of damaged skin after tape stripping

measured by TEWL values
70

60
Improvement (%)

50

40

30

20
Untreated Competitor Fameliocutin
 Post laser treatment

Improvement of skin irritation induced

by tape stripping
45

40
Improvement (%)

35

30

25

20
Untreated Competitor Fameliocutin
 Wound healing properties
Enhancement of TEWL after skin damage and
[ %] treatment with products
250
Market leader

200
Cream with
0.1% CM-Glucan
untreated
150

100

50

0
stripping day 2 day 4