AU700210B2 - Extraction of biological active organic silicon compounds of seaweed origin - Google Patents

Description

AUSTRALIA

Patents Act COMPLETE SPECIFICATION

(ORIGINAL)

Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: 444* t 4 t o t4 44 *4 I o 44 I 4 or 4.4.

44A 44 4 4I '4.4 I 4 0444 *40 4 f4 44 i Id 00 Name of Applicant: Exsymol Algues Et Mer Ocealys S.A.R.L.

Actual Inventor(s): Marie-Christine Seguin Andre Franco Emile Feno Jean-Yves Moigne Fabienne Bresdin Address for Service: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: EXTRACTION OF BIOLOGICAL ACTIVE ORGANIC SILICON COMPOUNDS OF SEAWEED ORIGIN Our Ref 445438 POF Code: 218871/277742,277768,277776 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1itn 7l 1 1 k z200 396 i 2 Technical field The present invention concerns compounds based on biologically active silicon and particularly an extraction process of biologically active silicon compounds from seaweed, the localization process of such compounds on specific parts of seaweeds and the culture process of the seaweeds used for the extraction of these compounds.

Previous state of the art Silicon, a widely present element in nature is mostly known under its natural inorganic forms such as silica and silicates or also as synthetic polymers silicones. These silicon compounds are slightly soluble or insoluble in aqueous medium which explains their small occurrence in living organism.

The works of the applicants have demonstrated that silicon compounds may constitute a form of silicon which can be assimilated by the body (by opposition to mineral silicon or to silicones), if they have the property of existing in aqueous solution in low molecular weight oligomers.

Moreover, an other necessary characteristic for the activity of those oligomers in aqueous solution is to exhibit numerous Si-OH function. Therefore it appears that the biological properties of those silicon compounds which can 25 be assimilated by the body, are only observed if they form, in a solution, soluble oligomers, coming from the linkage of i Si-O-Si siloxane bond, rich in Si-OH functions.

The chemical species implied in most of the above mentioned biological mechanism is a soluble form of silicon silicic acid, its formula being Si(OH)4. But knowing its strong ability to polycondensate for forming i silica, this compound has to be stabilized immediately after .the extraction.

One of the difficulties found by the applicants has been to obtain silicon based compounds in which silicon is

~_II_

3 linked by bonds which may be hydrolysed for obtaining biologically active compounds with one or many Si-OH functions. This kind of compound may of course be synthesised, but it is more advantageous to be able to extract directly these compounds from vegetals.

Unfortunately, all vegetals do not contain these type of silicon base compounds.

Statement of the invention The basic idea which prevailed in the birth of this invention is based on two statements. On one hand, there is a qualitative equivalence between the mineral content of human physiological liquid and the content of sea water, on the other hand, there is an analogy of cell multiplication mechanism between human embryo tissues and sea weed origin tissue.

That is why the major goals of the invention is to provide biologically active organic compounds from seaweed origin and to localize precisely which part of the seaweeds contained said biologically active organic silicon :compounds.

An other goal of the invention was to carry out the extraction from sea weed of biologically active silicon organic compounds.

25 An other goal of the invention is to carry out the cultivation of sea weed from which may be extracted biologically active silicon organic compounds.

The main purpose of the invention is therefore a biologically active silicon organic compound whose general S 30 formula is: R---Si-R 4 R3

EEP-

I i s;~~ns:rrr~lsl ~llll-nlllin which RI, R2, R3 and R4 are organic radicals, one of which at least is hydrolysable, of an type, X being a heteroatom and R' being H or an organic radical, said compound being a seaweed organs extract proceeding essentially from reproductive origin or young tissue.

An other purpose of the this invention is a method of localisation of the parts of the sea weeds from which may be extracted the compound according to the invention, including the following steps taken from a sample of the above mentioned sea weed parts fixation of the sample in a block of polymerized resin, slicing of fine cuts in the block of polymerized resin, then spectrophotometric micro analysis of one of those cuts for detection of which parts contained silicon at a concentration above a predetermined level.

An other purpose of the invention is an extraction method of organic silicon compound according to the invention including the steps consisting in causing the cell containing the compounds to be extracted to be blown up, in getting rid of thickening agents by precipitation, to :stabilising within the organic silicon compounds the naturally occurring Si-OH bonds or those obtained by hydrolysing the Si-OR' bonds, and in purifying the concentrates obtained by an ultra centrifugation 25 stabilisation.

An other purpose of the invention is a process of cultivating the sea weeds from which will be extracted the organic silicon compounds in which a sapling of seaweed whose cultivation is realized is sliced in many fragments so S" 30 as to form many microslips which will be immersed in a sea water filled tank, then a strech out shaped cultivation support such as a rope is placed in the tank so as for the 0 microslips to hook on the cultivation support and at the end, the cultivation support on which are hooked the I microslips is installed on a cultivation field in the sea, at a predetermined depth and parallel to the sea surface.

Brief description of the drawings The goals, targets and characteristics of the present invention will be better highlighted by reading the following description, being made in reference to the drawings in which figure 1 is a sketch of a harpoon branch of the sea weed Asparagopsis Armata in which some parts contain organic silicon compounds according to the invention.

figure 2 is a sketch of the fixed boat on which is installed the seaweed cultivation support before its installation in a sea cultivation field, figure 3 is a sketch of a roll based for passing the cord used as a cultivation support on a sea cultivation field, figure 4 is a sketch of a funnel bearing the casing enveloping the rope used as a cultivation support in the cultivation process according to the invention.

i Detailed description of the invention The applicants have found that certain part of macroscopic seaweeds contained biologically active organic 25 silicon compounds of general formula R-Si-R 4 ci in which Rl, R2, R3 and R4 are organic radicals at 30 least one of which is hydrolysable, of an type, X being a heteroatom and R' being H or an organic radical. The heterotaom being preferably an oxygen atom.

-I C 6 An original localization method, as described here under, proves that those silicon compounds are mainly located in the reproductive organs and or the young tissues of certain sea weed species.

LOCALIZATION METHOD Generally sea weeds contain silicon, whether as inorganic or as organic compounds. Therefore the occurrence of inorganic silicon is well known in unicellular sea weeds of diatomea type, in which the silicified shell is made by an extra cellular silicon deposit. But the applicants have discovered the organic compounds according to the invention are mostly inside the sea weed cells although improper silicon compounds, in the frame of the invention, have been localized in the intercellular cement.

Organic silicon compounds, the only interesting compounds in the frame of this invention, exist in variable quantity according to the sea weed species and are not found in every part of the sea weed. Therefore, certain species as those from the family of red algae or Rhodophyceas seem to be more interesting than others.

~It has been necessary to devise a method for localising the sea weed parts, and to determine which species will be selected for extraction of organic silicon compounds o .r 25 according to the above described method.

For one given sea weed variety, the preferred method of localization consist in setting the sea weed sample in a resin, then to slice the samples fixed in thin cuttings in 0order to realise a tissue microanalyse which consists in Xray spectrometric analyse followed by an ionic analyse.

To fix samples to the resin, the first step consists in 00 o- placing them in an usual fixative as glutaraldehyde or in formaldheyde-glutaraldehyde mixture in sea water solution, and a slight vacuum corresponding to a pressure less than 10 3 bar. Fixed samples can be post fixed by Osmium

I

7 tetraoxyde. During the second step, samples are dehydrated in successive baths with an increasing alcohol ratio. The use of sea water is better than pure water to maintain the same osmotic pressure as the natural medium of the cells. A third step consist in the inclusion of samples in a liquid thermohardening resin, under a little vacuum, in order to obtain a better diffusion of the alcohol and the resin ir cells, when the vacuum disappears. This technique induces step by step a substitution of alcohol by resin in samples.

The resin used is preferentially an epoxyde resin. During the last step, we realise the polymerisation of the resin more particularly by heating under a 70 *C.

The fixed samples are cut in thin layer (0,7 and semi thin layer (2 pL) on which the microanalyse is realised in order to select specifically the part of the algae which contain silicon organic compounds.

The micro analysis of fixed samples starts with an X Ray spectrometry assay on the thin cuttings (0,7 for the silicon detection in samples. This method gives the constitutive elements identification of the biologic tissue t. according to their specifics X bands. In this purpose, the sample is deposited on a copper bars. The sample is shelling by a thin beam of electron named "electronic sonding" with an energy between 0,1 to 50 Kev. The X ray spectra emitted 25 from the sample are analysed by an energy spectrometer.

Impulses are selected in function of their energy and an expert system interprets the results. With this technique, o.r all elements with an atomic number upper than 4 are °detected. The limit of this detection is around 10 14 to S 30 10-15 g, which corresponding to a concentration less than 100 ppm.

°If the analysed sample contains silicon, we proceed to an ionic analyse of the secondary ion, in order to localize the compartment of the sample in which silicon is. Because the secondary ionic analysis induces erosion of the 1 IIPLE~i~~~- cuttings, it will be preferable, for this technique, to use the semi thin cuttings (2 t) which results from the osmium tetraoxide post fixation.

The analyse of the secondary ions is a microanalytic method which combine the Mass spectrometry and the corpuscular microscopy. With the mass spectrometry, molecules are ionising by a chemical reaction. The different ions are separated in an analyser system in function of their ration between mass and charge The mechanism of the secondary ionic emission is obtained after the shelling of the samples by a primary ionic beam (the potential of this primary beam could be adjusted until about 12 kV). The primary ions strikes the sample's atoms and atoms which are next the surface. Samples stripped atoms are charged or in a neutral state. The charged particles (secondary ions) are used for the analyse. The secondary ions could have monoatomic structure or polyatomic structure. The corpuscular optic with bi-directional focusing gives a secondary ionic image filtered of the solid sample surface.

20 The specificity of this method is that it gives an 00PQ 0.6° identification of all of the elements of the periodic .00.

classification and an image of the distribution with a spatial resolution under one micrometer.

Unfortunately, the spectrometric mass analyses induce 25 artefacts and can be confounded between silicon and compounds with the same mass 28. This compound could be only Al-H+ because the others, like C2H4, N2, CO,CNH2, are usual 060: constituents of the biological compounds and give a diffuse image in the whole samples. It is necessary to realise an ionic analyse of Aluminium (Atomic mass 27) to reveal the parts of the sample containing Al. If the result is the same than the one obtained by the silicon analysis. It means that .it is the AL-H+ compound and not silicon. This sample must be thrown away. If there is no superposition of the results, 9 that means that, the located (during the micro analyse) part of sample really contains .ilicon.

The above method of localization allows the silicon localization in different parts of specific species of alga and the realisation of culture process in order to produce the parts of the alga which are the more interesting. Thus, in Asparagopsis Armata (Rhodophyceae), we have localised in an important way silicon species in the cystocarp (reproduction organ) and in apical tissue (young tissue), the localization of silicon is weak in the thalle and in the old branchea.

CULTURE PROCESS.

The organic compounds of silicon (according to the invention) possess a great economical interest which justifies the intensive culture of algae species and varieties from which we can extract them.

That's why an original culture process have been developed by the plaintiff according to the above described original culture (refer to figures 1 to 4).

Although the following process could be used for many o :species and is well indicated for Bonnemaisoniaceae family, and especially for Asparagopsis Armata specie.As a matter of fact, some algae like Asparagopsis Armata have harpoons 25 branches each Asparagopsis plants have 5 to 10 harpoons branches such as the branch 10 with its harpoons 12 on the 0 o*o picture 1. In the natural environment, branches which come On off by the sea mechanical action, are carried away by the 30stream and fix themselves on new support to induce, by this way, species propagation and perenniality.

The culture process uses the microslip which consists to cut every harpoon branches (with 10 to 20 harpoons by branches) in small parts of one or two centimetres. Every cutting of branch contains 1 or more harpoons. Thus supposing that there is no sterile fragments (which is very 1 difficult to avoid) it is possible to obtain a potential plant multiplication up to 50 to 200 times.

One of the essential characteristic of this process is to place microslips on a support before the immersion in the sea. In this way, we use a fixed boat (20) (illustrated in picture 2 where the tank full of sea water containing microslips Asparagopsis Armata is 5 litres of sea water for 1 Kg of microslips). A support 24 like a rope or a little net trackted by a moving boat (not shown) pass through this tank 22. Tile support 24 which is fit up in the culture field by the moving boat is choose preferentially shaggy and smoothless to fix microslips during its crossing in the bag 22. That's why a worn polyethylene rope is preferable to a new one. The diameter of the rope must be included between 25 to 50 mm to have an important surface for the development of numerous population of algae. The stock 26 of the rope (or net) is located in a tank of the stock 28.

The tank (or net 24) which undergo a traction power, because it is pulling by an extern one, goes though the 22 tank using 3 directing organs the first one (30) (to guide) is a simple cylinder tube made of smooth plastic which possess a free mobility around its axe. The second one (32) is an empty plastic tube setting freely on a tight 25 ball-bearing. This kind of rope (illustrated figure 3) has preferentially an hexagonal form and lines (34) which offers a limited contact with the rope number 24. This limited I contact is necessary to avoid taking off microslips from ropes that should certainly arrives if the directing organ 30 (32) was smooth. The third one (36) has the same shape as the one represented on the picture 3 in the same way as to avoid the disconnection of the microslips. But we can forecast that this directory organ (36) is driving in rotation (not free) in order to help the rope 24 to assure the crossing of the culture support in tank 22. It can be a F4 4 11 "Power block" type (hydraulic driving) with spur surface to reduce the contact of the culture support.

We must notice that the culture support number 24, (rope or net) must be first wet with sea water in order to avoid mop up of sea water contained in the tank 22. In order to do so, a solution consist in keeping the rope stock in the sea water which fills the stocks 28.

An enveloping sheath must be deposed around the culture support by the outlet device (40) (see the detailed figure 4 so as to avoid that the cuttings go away from the support when being submerged by sea. Such a device may be realized with a fine-mesh cotton net (1/10 mm). Its reserve 42 surrounds a polyvinyl chloride curved-ends sleeve or spout 44. The sheath 46 is pil led by a moving boat (not shown) in the same way as the rope 24. When going away from the sleeve 44, the sheath 46 tends to draw nearer to the rope and finds itself close to it owing to the fact that the meshes are tightening lengthwise. So as to obtain a right tension of the sheath 46 et then a reduction of its diameter to get a right adhesion to the rope 24, the spout 44 outlet o.a is equipped with a brake 48 made of a rubber ring.

o The sheath 46 is meant to maintain the hooking of the umicrocuttings to the culture support. It should be noted 6 :00 that it is made of "degradable" matter so as to be 00. 25 decomposed slowly into seawater in 10 to 20 days, the sufficient time to allow the fixing of the microcuttings on a the support without obstructing their growth beginning.

ij i Then the culture support or rope 24, surrounding with its sheath, is submerged in a culture field, at an optimal depth of about 1 meter to allow a good penetration of daylight. The supports are maintained parallel to the g surface at these depth by a floats set. In a culture field (a 100 meters-sided square), the culture supports in groups of four are separated by a plastic (PVC) barrs set equipped with floats. Therefore such a device allows to accommodate 12 up to 80 culture supports 50 meters in length, that is to say 4 km of culture supports for a 100m x 100m field.

Even though the preferred support for this culture process is made of a continuous rope passing through a container of microcuttings, variable devices are possible without going beyond the limits of the invention. So instead of using a continuous rope, it is possible to use ropes lengths (10, 20 or 50 meters) which are submerged into the container of microcuttings before being set up in a culture field.

As seen previously, the organic compounds of silicon are localized in the young organs of the alga. So owing to this fact and because the alga is able to produce new branches quickly after a first cut, the culture will be executed by successive crops spaced 1 month apart. So most of new branches rich in organic compounds of silicon will be harvested. This process of monthly crop of branches will be possible to use quite all the year round. Moreover the biological evolution of the algae leads to produce reproductive organs yearly which will be ready for picking 1° in July and August.

EXTRACTION PROCESS C The localization method as described previously, allows S 25 to select the algae species which are interesting for

CC.,

extraction of organic compounds of silicon according to the invention, as well it allows a selective extraction of the organic compounds of silicon from the alga parts the most 0* 30 interesting according to this extraction.

To realize the extraction of the organic compounds of silicon, it is essential to burst the cellular walls of the alga. 2 methods can be used the cryopulverization or the SI. C ultra-high pressure.

The cryopulverization consists of a previous freezing of the algae by 5-6 kg sheets. These sheets are introduced 13 into a crusher where liquid nitrogen is injected so as to obtain a compound at minus 50 degrees Celsium finally. This compound is particle-shaped, in a vacuolar form (the particles are separated by wide spaces) and therefore voluminous since the cryopulverization has led to a bursting of a great number of cells and consequently a release of intracellular content. It shall be noted that other techniques can be used to induce the cells bursting.

With ultra-high pressure, the algae are put through pressures ranging from 4 000 to 10 000 bars during 6 to mn. So every alga cell undergoes a isostatic and omnidirectional high-pressure leading to its bursting.

As soon as the cell walls are bursted, it is essential to precipitate the alga thickeners the carrageenans) by ammonium -like compounds or every other process Even if the organic compounds of silicon are stabilized partly by natural stabilizers from the alga, this stabilization has to be completed by using of these natural stabilizers preferably. This stabilization results of the creation of weak linkages (hydrogen bonds) preventing the S4. polycondensation according to the Si-OH linkages. The used So stabilizers could be carboxyl hydroxyacid compounds, o particularly the a- and b- hydroxyacids, the glucuronides, S: o hydroxyl or phenol aminoacids, compounds with many alcohol (or phenol) groups. In this class we mention the glycols, the catechols and the catecholamines, the polyalcohols such S.as the glycerol, the monosaccharides or phenol acids such as gallic acid.

o After the stabilization, the concentrates of organic compounds of silicon are purified at the end by ultracentrifugation (very-full-speedy centrifugation) and dialysis. The silicon by-products other than the organic compounds wanted are eliminated within the centrifugation residues. In case of the aforesaid species, the silicone 14 content of the solution obtained with this extraction method was about 0.3 g/l.

APPLICATIONS

In general, the compounds extracted with the aforesaid method will be applied in many ways owing to the fact that they have the properties of the organic cilicon forms (therapeutic, dietetic and cosmetic properties) following from their anti-inflammatory, hydrating, tissues regenerative, anti-degeneration, normalizing, lipolytic, metabolic stimulating, anti free radical or anti-glycation activity, or in a more general way an activity of stimulation of the organisms defenses.

So the further on examples illustrate the activity of the organic compounds of silicon extracted from algae in cosmetology (ex.1) and in dietetics (ex. 2, 3 Example 1 (dietetic therapy) Capsules have been made. One capsule contains 0.59 g of 20 organic compound of silicon extracted from algae as the o m« o invention principles in a pulverulent form (the extract is S• adsorbed on a polyamide powder) that is to say 0.1 mg of o :silicone per capsule.

The posology was 6 to 9 capsules per day. The daily o 25 absorption of algal silicon set back in good form with reeducation or disappearance of bone pain, the treatment o .having to be continued for several months.

00O Example 2 (tissue regeneration activity) 0. 30 We have carried out a cell culture of human fibroblasts in microtiter plates. 8 ml of a titrated solution of organic 00 g silicon compound from algal origin was diluted in one litter of MEM medium added with FCS 2,5 and distributed in the wells.

A

5' I a U Two control series were made containing 10 FCS. Cell multiplication is monitored using the Neutral Red coloration technique (Borenfreund 1984). The dye-wavelength absorbance measurement is directly proportional to the numiber of living cells. The cell multiplication was increased of 55 in the presence of algal organic silicon.

Example 3 (moisturizing activity) We have made an aqueous gel containing 12 of algal organic silicon obtained according to the invention. This gel was applied on the right foreharm for 3 days. The moisturizing effect was measured by a Fourrier Transformed Infra Red technique, the hydration degree being given by the ratio of the surface of the amide 1650 cm- 1 absorption band to the surface of the amnide 1550 cm-I(C-N) absorption band. We have measured daily thi. hydration degree, between the 4th and the 10th day, and compared it to a reference gel without silicon applied on the left foreharm.

This test was carried out on six persons. The mean values 20 are the followings *0 at D 1 after application, we measured an hydration degree of 2,3 as compared to a degree of 1,5 for the control.

at D 7 That hydration degree reached 2,09 while 1,37 for the control.

These results reveal well the immediate effect and 4: moreover a time-lasting effect, of an evident biological moisturizing activity (bound water).

Example 4 (lipolytic activity) oWe have maintained in a survival state human adipocytes. We added 10 pil of a solution of algal organic 4 0 silicon (containing 30 mg per litter of silicon) diluted in 100 jil of NH4HCO3 0,01 M, and completed to 3 ml with a Krebs-ringer buffer.

16 The lipolytic activity was quantified by the measurement of the release of glycerol using an enzymatic method. Results were reported in nanomoles of glycerol per grams of total lipids. The algal silicon-containing compound at the chosen dose can stimulate basal lipolysis between 0 and 60 mn. The improvement is 134 as compared to the control.

1 a j Th:ioy ciiy a uniid b h o B

Claims (17)

1. Organic silicon compound biologically active having as a general formula: R9 I RL-Si-R 4 C a a* Co i o r o a I 4 i€0 in which R1, R2, R3, and R4 are organic radicals with at least one of them being hydrolysable, of the type where X is an hetero-atom and R' being an atom of hydrogen or any organic radical; wherein said compound is an extract of algal origin coming essentially from the reproductive organs and the young tissues of the alga.

2. Compound according to claim 1 in that said heteroatom is the atom of oxygen.

3. Method of location of the algae parts from which can be extracted the compound according to claim 1 or 2, including the following steps preformed on a sample of the said parts of the algae: fixation of the said sample in a block of a polymerized resin slicing in fine cuts of the said block of a polymerized resin, and elemental analysis of the said fine cuts by spectrometry in order to locate the parts of the algae having a content of silicon superior to an already chosen level.

4. Method according to claim 3, in which said fixation step of the said sample of alga in the polymerized resin includes the following steps: fixation of the said sample by the means of a fixative such as glutaraldehyde, and under a slight vacuum dehydration of the said fixed sample by successive bathing in a series of alcohol-containing bath of increasing degree of alcohol, *C a C \WHODPUA\PCeu( e DO I a ao. a a Ia..a a 1~ 18 placing said dehydrated sample in a non polymerized resin in such a way that said resin can replace slowly the alcohol contained in the said sample, and proceed to the polymerization of the resin.

Method according to claim 3 or 4, in which the said step of elemental analysis consists in: detect if the analysed sample contains some silicon using x ray spectro metry, carry out an ionic analysis of the secondary ions in order to visualize the parts of the sample where silicon is located, and withdraw the said parts from the rest of the sample when they are the same that the one identified by ionic analysis of aluminium.

6. Procedure for the culture of the alga from which is extracted the organic silicium-containing compound according to the claim 1 or 2, wherein it includes the following steps: the algal sapling to be cultured is cut into fragments in order to obtain as many microslips, and said microslips are immersed in a tank filled with sea water, a long-shape culture rest such as a rope is dipped in the said tank in such a way that the microslips can fix to the said culture rest, and said culture rest on which are fixed the said microslips, are then placed in a culture field in open sea at a predetermined depth and kept parallel to the water surface.

7. Culture procedure according to the claim 6, in which the said tank containing the said microslips is a fixed boat and the said culture rest is a continuous rope pulled by a moving boat being forced to remain immersed in the said container as it progressively induces the microslips to fix to the rope.

8. Culture procedure according to the claim 6, in wNhich said rope is shaggy in order to allow a good fixation of the said microslips on the said rope.

9. Culture procedure according to one of the claims 6, 7 or 8, in which the said culture rest is covered with a biodegradable wrapping sheath before being place in the said culture field.

Culture procedure according to any of the claims 6 to 9, in which the said algal sapling is choosen among algae that are bearing harpoon-like twigs, C %WNWOR"AULAPiECIES85996 DC t k- 19 each of the said microslips presenting at least one harpoon thus allowing an easy fixation on the culture rest.

11. Culture procedure according to claim 9, in which the said alga bearing harpoon-like twigs belongs to the species Asparagopsis armata.

12. Extraction procedure of the said organic silicon compound according to the claim 1 or 2 consisting in: induce the burst of the cells in the parts of the algae that contain the said compounds, withdraw the thickening agents of the alga by selective precipitation using ammonium-type agents, stabilize in the said organic silicon compound, the natural Si-OH linkages or linkages induced by the hydrolysis of the linkages Si-OR', and purify the concentrates resulting of the stabilization procedure by ultra- I centrifugation. 15

13. Extraction procedure according to the claim 12, in which the burst of the cells in the parts of the alga that contain the said organic silicon compound, is obtained by cryopounding at about -50 0

14. Extraction procedure according to the claim 12, in which the burst of 0 0 the cells in the parts of the alga that contain the said organic silicon compound, is oo So 20 obtained by ultra high pressure, comprised between 4,000 and 10,000 bars.

15. Application of the organic silicon compound according to the claim 1 0 or 2, in order to obtain a therapeutical, dietary or cosmetic composition, having an i 2" 5 anti-inflammatory tissue regenerative, normalizing, lipolytic, metabolic stimulating, anti-free radical or anti-glycation activity, or in a more general way an activity of stimulation of the defense mechanisms of the human or animal organisms.

16. A compound according to claim 1 substantially as hereinbefore described with reference to any of the examples.

17. A procedure according to claim 6 substantially as hereinbefore described with reference to any of the examples. CA\WINWORDWICHELLe1NODELTE\SPECIES\4815A-96DOC 4 0 444 4 o 44 DATED: 30th June, 1998 PHILLIPS ORMONDE FITZPATRICK Attorneys for: EXSYMOL ALGUES ET MER S.A.R.L. and OCEALYS S.A.R.L. C IVANWORO\PAULASPECIES\43lSQ .g4DOC 44 I 9