05-BoNtA 568

BoNtA 568
Lets relax not paralyse muscles
Dr Philippe Deprez
Policlnica Esttica & anti aging National academy of Sciences Bucuresti Honour Member
Port Grec, 40, pb 17487 Empuriabrava - Espaa

Tel +34 972 45 01 51 Fax: + 34 972 45 23 23 http://www.estetik.com/centres/cmeempur.htm phd@estetik.com 1
What means BoNtA 568 ?

BoNtA is the acronym of Botulinic Neurotoxin Alpha
568 represent the number of Amino acids in the BoNtA-like oligopeptides contained in the product 5- pentapeptide 6- hexapeptide 8- octopeptide
Aims of the treatment

Botulinic toxin and BoNtA 568 have the same target: Interfere with Acetylcholine (Ach) synthesis and release in order to control muscle contraction
Is BoNtA 568 similar to
Botulinic toxin injection ?
Other Botox-like products of the market ?
BoNtA 568 www.aestheticdermal.com
Is BoNtA 568 identical to a Botulinic toxin ?

NO
Botulinic toxin
Destroys the axone that grows back in 3 to 4 months Totally blocks Ach Release in the synaptic clefts Induces a total flaccid palsy of treated area
BoNtA 568
Does not destroy axone Slows down Ach release in synaptic cleft Induces a transitory muscle relaxation
BoNtA 568 www.aestheticdermal.com 5
Is BoNtA 568 similar to other botox like products of the market ?
NO
Usual Botox-like products
Use only 1 oligopeptide (usually hexapeptide) In a low concentration ( 5 or 6% usually)
BoNtA 568
Uses 3 synergetic oligopeptides 3 x 20 % concentration in Medical Care 3 x 10 % concentration in Daily Care
BoNtA 568
Does not paralyse muscles like Botulinic toxin Uses a synergetic combination of 3 botox like oligopeptides in a much more concentrated way comparing to any other product of the market.
BoNtA 568 presentations

MEDICAL CARE Phials
To be injected (CE pending) No Needle mesotherapy use
DAILY CARE gel

2 times / day during the first 2 weeks Once daily as daily care
Efficacy
In Vivo silicone imprints ( around the eyes ) after topical application
D0
D 28 topical pentapeptide contour eyes
Blue = deep Red = High
D0
D 28 topical SNAP 6 contour eyes
10
DO
D 28 topical Actyl-hexa + Penta peptides Contour eyes
Blue = deep Red = High
DO
D 28 topical Snap 8 contour eyes
11
Efficacy has been proven clinically and experimantally

1. 2. Skin wrinkles silicone imprints Studies on glutamate neurotransmitter release in vivo Electrophoreses of SNARE complex and SNAP 25 protein Studies on catecholamine exocytosis with and without oligopeptides BoNtA like Studies on action of pentapeptide on external axonal enkephaline-like receptors Monitoring of Snare complex formation
Evidences 2 to 6 need more extensed explanation
3.
4. 5. 6.
There is no toxicity
-cytotoxicity: human keratinocytes and human fibroblasts : NO -Ames test (geno toxicity) . NO -Eyes irritation text /HET-CAM test (Hens Egg Test-Chorio Allantioc Membrane)
and NRU: (Mouse Fibroblast Neural Red Uptake TEst): NO EYES IRRITATION
-LD50 rats: > 2.500 mg/kg = strictly NO TOXICITY -Occlusive patch tests on human skin : NOT IRRITATIVE -Intraepidermal abrasion + occlusive application: NOT IRRITATIVE (0 / max score 8) -HRIPT (Human Repeated Insult Patch Test) -Hemolysis test : NOT HEMOLYTIC -Subcutaneous injection (SNAP6) rabbits: Extremely low irritating power ( 0.55 on a 0 to 8 score) -Intraperitoneal injection (snap 6): 1-100 mg/kg: NO TOXICITY
13
3 ways to use BoNtA 568
Classical Mesotherapy
(CE as medical device pending)
Botox prolongator
No needle mesotherapy
BoNtA 568 Injections

as a skin relaxing anti wrinkles treatment
INJECTIONS
Inject more superficially compared to Btx Do 2 times more injection points

NOTES
Meso botox is less efficient than classical botox, except if similar doses are used BoNtA-like 5-6-8 will have the same level of efficacy than low doses meso botox BoNtA-like 5-6-8 are absolutely not toxic CE for injection pending
NO PALSY will occur, the skin will be relaxed, no risk of blepharochalasis, or eyebrows ptosis No risk of headache or other side effect linked to botulinic toxin injection
BoNtA 568 daily care as Botulic toxin prolongator

Botulinic toxin is injected as usual for blocking completely the muscles BoNtA 568 daily care is applied every day to prolong the result (30% maximum)
Transcutaneous penetration of BoNtA 568

Botulinic Neurotoxin A
Skin is virtually impermeable to this macromolecule BoNtA is sensitive to oxidation and would not survive long on the surface of the skin
BoNtA-like 5-6-8
Low sensibility to oxidation: survive on the surface of the skin Low molecular weight: simple transcutaneous penetration has been proven (Franz Cell)
Franz Cell : penetration of hexapeptide through human skin

Chamber 1
Chamber 2
After 2H: 36% of chamber 1 passed in chamber 2 After 4H: 48% Ch 1 After 6H: 78% Human Skin After 24H: 90%
Ch 2
2-3 mm thick human skin
17
Trans epidermic penetration

AD ROLL or Superficial abrasion dramatically increase skin permeability
Products penetrate into the dermis . Is it true ? Evidences: -Application of adrenalinated lidocaine after brasion -Or after AD roll -Induce -Anaesthesy: Nerves termination are located inside of deeper than the epidermal basal layer -Vasoconstriction : Blood vessels are located inside of papillary and reticulary dermis. In the same time, vasoconstriction slows down vascular resorption
BoNtA 568 No Needle mesotherapy

a skin relaxing anti wrinkles treatment
3 steps no needle meso technique
1- skin permeabilization Skin abrasion ( AD abrasive kit) Or ROLLER ( AD ROLL)
2- Product application Apply the product on the skin Cover using a kling film ( Avoids Evaporation) Let the active products penetrate into the dermis
3- Induce an Intracellular penetration for avoiding vascular fast resorption EXCELLDERM
In short, about no needle meso

1- disinfect the skin 2- permeabilize the skin AD roll (Aesthetic Dermal) Or ABRASION (Aesthetic Dermal) 3- apply the active products on the skin BoNtA 568 as unique product
Or
More complex mixture ( see later) 4- cover with kling film . Intradermal penetration 5- intracellular penetration: Excellderm . Action every day
6- repeat 2 times month ( 4 sessions total) and use BoNtA daily care
How oligopeptides works
The detailed science under the cosmetic product
1- Action potential
Simplified Mechanism of Muscle contraction

2- depolarisation
BoNtA & BoNtA 568 act on membranes fusion phenomenons
axone
3- SNARE complex formation 4- attraction of Ach vesicle to axone membrane 5- Membranes
ACh Vesicle
6- liberation of ACh in synaptic cleft 7- activation of muscle receptors
fusion
Synaptic Cleft
Muscle
8- Muscle contractions =>wrinkles
Actors playing a role in membrane fusion

2x4 proteins are needed for membrane fusion
Axone
Neurotransmitter
Muscle
Vesicle containing Neurotransmitter Phospholipids bilayer
V-SNARE = VAMP protein = Vesicle Associated Membrane Protein
Goal can This is: neurotransmitter be done thanks to has membrane to pass through fusion porosome mechanism
SNAP-25 SyNaptosomal Associated Protein
T-SNARE= Target SNARE Syntaxin Axone POROSOME
Axon Synaptic membrane Phospholipids bilayer
23
Nerve Action potential induces combination of proteins

Axone
Neurotransmitter
Muscle
Vesicle containing Neurotransmitter Phospholipids bilayer
4 proteins will combine in a

TORSION HELIX
Building the SNARE complex

T-SNARE= Target SNARE Syntaxin (vue dartiste)
SNARE = SNAp REceptor SNAP-25 or SyNaptosomal Associated Soluble N-ethylaleimideProtein sensitive fusion factor Attachment proteins REceptors
Synaptic membrane Phospholipids bilayer
24
Nerve Action potential: combination of proteins

And calcium flux into the axon.
This is SNARE complex
In response to a motor action potential, the 3 proteins (VAMP- Syntaxin and SNAP 25) combine and form a four helix protein complex called SNARE
This combination captures the neurotransmitter-filled vesicle, attract it and dock them very close to the synaptic membrane
But there is still water molecules [Ca(H20)n]2+ between the 2 bilayers and fusion is not possible as it BoNtA 568 www.aestheticdermal.com 25
= water = Ca-P bridge
Thanks to proteinic helix torsion forces, vesicle membrane and axone bilayer membranes are brought to within 2-3 Angtrms of each other. This allows calcium phosphate bridges between the 2 (-) surfaces Vesicle-axonal plasmatic membrane that usually repel each other [Ca(H20)n]2+ is more than 6 Angstrms => is physically explused
= water = Ca-P bridge
Ca-P bridges simply expel the water still separing vesicle membrane and
axon plasmatic membrane The little space left now between the 2 bilayers also displaces water bound to the P group of phospholipids => membrane destabilization and fusion
Membranes fusion
Synaptic cleft
Muscle membrane receptor Ach sensitive
Opening cation channel => muscle contraction
28
How can BoNtA and BoNtA-like oligopeptides Interfere with this mechanism ?
All serotypes of Botulic Neutotoxins type A (BoNtA) are known polypeptides 2 PHASES OF ACTIVATION ARE NEEDED FOR A NEUROTOXICITY
BoNtA is a folded polypeptide chain, linked by a disulfure bond.
Heavy chain and light chain

As it: Not toxic Neurotoxicity needs 2 structural modifications 1st = cleavage between 2 amino acids 448-449 ? 418-419? 422-423?
30
2nd phase of BoNtA activation

This cleavage allows VERY RAPID entry of BoNtA into axon (endocytosis)
Into the axon: 2nd modification: Disulfure bond is cut
31
BoNtA lyses SNAP 25 protein

Heavy and light chains are liberated LIGHT CHAIN IS A METALLO PROTEASE That lyses SNARE PROTEINS
32
BoNtA lyses SNAP 25 protein

Botulinic Toxins Light chain, a Zn dependant endoprotease lyse VAMP i.e.
SNARE is impossible MEMBRANE FUSION is impossible Ach release is impossible
BoNtA lyses SNAP 25 And kills the axone
Muscle contraction does not occur
X
BoNtA destroys axon

BoNtA destroys axon Axon regenerates by sprouting Sprouting begins within the first few days after BoNtA has been injected It takes 10 20 weeks for the muscle mobility to be completely regained thanks to the formation of new synapses BoNtA-like oligopeptides 5-6-8 have other modes of action on SMARE complex formation and do not destroy the synapse.
Oligopeptides BoNtA like

Will mimick this action mode
What are BoNtA - like oligopeptides ?
The actual trend to use transdermal therapies without injections has led to the creation of new products derived from BoNtA: oligopeptides BoNtA like
They do not block the muscle completely as do BoNtA but, acting at the level of SNARE complex, they MODULATE muscle contraction, giving a relaxation effect
What types of peptides are used and how do they work ?
36
Types of peptides
Pentapeptide 3 (5 amino acids) acts as an inhibitory enkephalin Acetyl Hexapeptide 3 (6 amino acids) competitor for SNAP 25 protein Octopeptide or SNAP-8 (8 amino acids) competitor for SNAP 25 protein
=> The name oligopeptides BoNtA Like 5-6-8)
Pentapeptide 3 ( 5 amino acids)
Pentapeptide
Pentapeptide 3 is a modified enkephalin that couples to enkephalin receptors on the outer membrane of axons, that are coupled to a Gi protein (inhibitory G protein). Stimulation of Gi receptors => Ca channel are closed and K channels are opened. Membrane fusion is a process strictly Ca dependent BoNtA 568 38 => less ACh exocytose. =>www.aestheticdermal.com REDUCTION OF NEURON EXCITABILITY
Acetyl Hexapeptide 3 and Octopeptide (SNAP8)

Acetyl Hexapeptide 3 (SNAP 6) is similar to the last 6 amino acids of N-terminal of SNAP 25 protein Octopeptide (SNAP-8) is an elongation of acetylhexapeptide 3
SNARE cannot be stable with such a short polypeptide
SNAP 6 or 8
Competition in SNARE complex formation
Axone POROSOME
SNARE complex destabilized by SNAP-8 or SNAP 6 reemplacing SNAP 25

Less exocytosis Muscle relaxation
Efficacy of BoNtA like oligopeptides Glutamate release monitoring

Glutamate is the most common brain neurotransmitter and its essay is used to estimate that of ACH (membrane fusion phenomenons are constant)
Synergy HEXA + PENTA
40
Efficacy of BoNtA like oligopeptides Neuronal exocytosis and catecholamines release

Studies carried at University Miguel Hernandez, Alicante , Spain
L glutamate release after depolarization of cultured neurons: Hippocampus cells of rats embryos
Acetyl Hexapeptide-3 (SNAP6) and SNAP 8

Are more effective than Pentapeptide 3 But Pentapeptide is synergic with SNAP 6 and SNAP 8
Exocytosis of catecholamines from chromaffines cells monotor release of catecholamins (adrenalin noradrenalin)
SNAP 8 is also more effective than Snap 6

Efficacy of BoNtA like oligopeptides SNARE complex formation monitoring
SNARE complex cannot be formed correctly in presence of SNAP 6 or SNAP 8 (proteins electrophoresis) SNARE complex formation in vitro
Efficacy of BoNtA like oligopeptides SNARE complex formation monitoring
SNAP 8 (octopeptide) is more effective than SNAP6 (Acetyl Hexapeptide 3) in SNARE formation inhibition in vitro
43
Thank you
For Your attention
Policlnica Esttica & anti aging
Dr Philippe Deprez
National Academy of Sciences - Bucuresti Honour member
Ed dunitz English version (450pp) available february 2007 Korean version (600pp): available septembre 2006 See on www.estetik.com
Port Grec, 40, pb 17487 Empuriabrava - Espaa

Tel +34 972 45 01 51 Fax: + 34 972 45 23 23 http://www.estetik.com/centres/cmeempur.htm phd@estetik.com 44

05-BoNtA 568

Cargado por

Información del documento

Descripción original:

Derechos de autor

Formatos disponibles

Compartir este documento

Compartir o incrustar documentos

Opciones para compartir

¿Le pareció útil este documento?

¿Este contenido es inapropiado?

Copyright:

Formatos disponibles

05-BoNtA 568

Cargado por

Copyright:

Formatos disponibles

BoNtA 568

Lets relax not paralyse muscles

Port Grec, 40, pb 17487 Empuriabrava - Espaa

What means BoNtA 568 ?

Aims of the treatment

Is BoNtA 568 similar to

Botulinic toxin injection ?

Other Botox-like products of the market ?

BoNtA 568 www.aestheticdermal.com

Is BoNtA 568 identical to a Botulinic toxin ?

Is BoNtA 568 similar to other botox like products of the market ?

BoNtA 568 www.aestheticdermal.com

BoNtA 568 presentations

DAILY CARE gel

BoNtA 568 www.aestheticdermal.com

In Vivo silicone imprints ( around the eyes ) after topical application

D 28 topical pentapeptide contour eyes

Blue = deep Red = High

D 28 topical SNAP 6 contour eyes

D 28 topical Actyl-hexa + Penta peptides Contour eyes

Blue = deep Red = High

D 28 topical Snap 8 contour eyes

Efficacy has been proven clinically and experimantally

3 ways to use BoNtA 568

BoNtA 568 Injections

Inject more superficially compared to Btx Do 2 times more injection points

BoNtA 568 daily care as Botulic toxin prolongator

Transcutaneous penetration of BoNtA 568

Franz Cell : penetration of hexapeptide through human skin

2-3 mm thick human skin

BoNtA 568 www.aestheticdermal.com

Trans epidermic penetration

BoNtA 568 No Needle mesotherapy

1- skin permeabilization Skin abrasion ( AD abrasive kit) Or ROLLER ( AD ROLL)

3- Induce an Intracellular penetration for avoiding vascular fast resorption EXCELLDERM

In short, about no needle meso

How oligopeptides works

The detailed science under the cosmetic product

Simplified Mechanism of Muscle contraction

BoNtA & BoNtA 568 act on membranes fusion phenomenons

Actors playing a role in membrane fusion

Vesicle containing Neurotransmitter Phospholipids bilayer

V-SNARE = VAMP protein = Vesicle Associated Membrane Protein

T-SNARE= Target SNARE Syntaxin Axone POROSOME

Axon Synaptic membrane Phospholipids bilayer

BoNtA 568 www.aestheticdermal.com

Nerve Action potential induces combination of proteins

V-SNARE = VAMP protein = Vesicle Associated Membrane Protein

Vesicle containing Neurotransmitter Phospholipids bilayer

4 proteins will combine in a

Building the SNARE complex

Synaptic membrane Phospholipids bilayer

BoNtA 568 www.aestheticdermal.com

Nerve Action potential: combination of proteins

= water = Ca-P bridge

= water = Ca-P bridge

Opening cation channel => muscle contraction

BoNtA is a folded polypeptide chain, linked by a disulfure bond.

Heavy chain and light chain

BoNtA 568 www.aestheticdermal.com

2nd phase of BoNtA activation

Into the axon: 2nd modification: Disulfure bond is cut

BoNtA 568 www.aestheticdermal.com