NZ286213A

NZ286213A - Organo-silicon compound having at least one hydrolysable group being an extract of reproductive organs and/or young tissues of algae, culturing algae in sea water, method of extraction of organo-silicon compounds

Info

Publication number: NZ286213A
Application number: NZ286213A
Authority: NZ
Inventors: Marie Christine Seguin; Andre Franco; Emile Feno; Jean-Yves Moigne; Fabienne Bresdin
Original assignee: Exsymol Sa; Algues Et Mer Sarl; Ocealys Sarl
Priority date: 1995-03-22
Filing date: 1996-03-19
Publication date: 1997-12-19
Also published as: EP0733636B1; DE69619148D1; CA2172226A1; AR002037A1; FR2732022A1; AU700210B2; BR9601105A; EP0733636A1; FR2732022B1; AU4815996A; KR960034208A; ES2172648T3; JPH0931079A; DE69619148T2

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number £86213 New Zealand No. 286213 International No. PCT/ TO BE ENTERED AFTER ACCEPTANCE AND PUBLICATION Priority dates: 22.03.1995; Complete Specification Filed: 19.03.1996 Classification:^) C07F7/02.04; C12P9/00; A61K7/48; A23L1/325.48 Publication date: 19 December 1997 Journal No.: 1423 NEW ZEALAND PATENTS ACT 1953 complete specification Title of Invention: Extraction of biological active organic silicon compounds of seaweed origin Name, address and nationality of applicant(s) as in international application form: EXSYMOL S.A.M., company of 4 Avenue Prince Hereditaire Albert, 98000 Monaco, France; ALGUES ET MER S.A.R.L., a French company of Kernigou> 29242 Me D'Ouessant, France; OCEALYS S.A.R.L., a French company of Technopole de Brest Iroise, 29200 Brest, France 286 213 NEW ZEALAND PATENTS ACT, 1953 No: Date: COMPLETE SPECIFICATION EXTRACTION OF BIOLOGICAL ACTIVE ORGANIC SILICON COMPOUNDS OF SEAWEED ORIGIN We, EXSYMOL S.A.M., a Monegasque company, 4 Avenue Prince Hereditaire Albert, 98000 Monaco, France, ALGUES ET MER SA.R.L., a French company, Kemigau, 29242 He D'Ouessant, France and OCEALYS S.A.R.L., a French company, Technopole de Brest Ircise, 29200 Brest, France, do hereby declare the invention for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement 2 8 6 2 1 3 Technical field The present invention relates to compounds based on biologically active silicon and particularly an extraction process of biologically active silicon compounds from 5 seaweeds, 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. State of the art 10 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 15 living organisms. 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 20 existing in aqueous solution in low molecular weight oligomers. Moreover, an other necessary characteristic for the activity 6f those oligomers in aqueous solution is to exhibit numerous Si-OH function,, Therefore it appears that the biological properties of those silicon compounds which 25 can be assimilated by the body, are only observed if they form, in a solution, soluble oligomers, coming from the linkage of 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 30 silicon : silicic acid, its formula being Si(OH)4. But knowing its strong ability to polycondensate for forming silica, this compound has to be stabilized immediately after the extraction;. One of the difficulties found by the applicants has 3 2 8 6 2 13 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 5 extract directly these compounds from vegetals. Unfortunately, all vegetals do not contain these type of silicon base compounds. Summary of the invention 10 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 15 multiplication mechanism between human embryo tissues and sea weed origin tissue. That is why the major object of the invention is to provide biologically active organic compounds from seaweed origin and to localize precisely which part of the 20 seaweeds contained said biologically active organic silicon compounds. Another 6bject of the invention is to carry out the extraction from seaweed of biologically active silicon organic compounds. 25 Another object of the invention is to carry out the cultivation of seaweed 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 30 formula is : r2 I - Rj—-Si—R4 R3 °' : c 4 28 62 13 in which Rl, R2, R3 and R4 are organic radicals, with at least one of which being hydrolysable, of an -X-R' type, X being a heteroatom and R' being H or an organic 5 radical, said compound being an extract of algal origin coming substantially from the reproductive organs and the young tissues of the alga. Another purpose of the this invention is a method of localisation of the parts of the sea weeds from which may 10 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 15 analysis of one of those cuts for detection of which parts contained silicon at a concentration above a predetermined level. Another purpose of the invention is an extraction method of organic silicon compound according to the 20 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 25 hydrolysing the Si-ORV bonds, and in purifying the concentrates obtained by an ultra centrifugation stabilisation. Another purpose of the invention is a process of cultivating the sea weeds from which will be extracted the 30 organic silicon compounds in which a sapling of seaweed whose cultivation is realized is sliced in many fragments so 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 35 so as for the microslips to hook on the cultivation 2 8 6 213 support and at the end, the cultivation support on which are hooked the microslips is installed on a cultivation field in the sea, at a predetermined depth and parallel to the sea surface. 5 Brief description of the drawings The objects, purposes and characteristics of the present invention will be better highlighted by reading the following description, being made in reference to the 10 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 15 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, 20 figure 4 is a sketch of a funnel bearing the casing enveloping the rope used as a cultivation support in the cultivation pfocess according to the invention. Detailed description of the invention 25 The applicants have found that certain parts of macroscopic seaweeds contained biologically active organic silicon compounds of general formula : Rz i Rt—Si—R4 R3 30 in which Rl, R2, R3 and R4 are organic radicals at least one of which is hydrolysable, of an -X-R' type, X 286213 6 being a heteroatom and R' being H or an organic radical. The heterotaom being preferably an oxygen atom. An original localization method, as described here under, proves that those silicon compounds are mainly 5 located in the reproductive organs and or the young tissues of certain sea weed species. LOCALIZATION METHOD Generally sea weeds contain silicon, whether as 10 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 15 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 20 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. 25 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 according to the above described method. For one given sea weed variety, the preferred method 30 of localization consist in setting the sea weed sample in a resin, then to slice the samples fixed in thin cuttings in order to realise a tissue microanalyse which consists in X-ray spectrometric analyse followed by an ionic analyse. r* ] 7 286213 To fix samples to the resin, the first step consists in placing them in an usual fixative as glutaraldehyde or in f ormaldheyde-glutaraldehyde mixture in sea water solution, and a slight vacuum corresponding to a pressure 5 less than 10~3 bar. Fixed samples can be post fixed by Osmium 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 10 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 in cells, when the vacuum disappears. This technique induces step by step a 15 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 m) and 20 semi thin layer (2 m) on which the microanalyse is realised in order to select specifically the part of the algae which cbntain silicon organic compounds. The micro analysis of fixed samples starts with an X Ray spectrometry assay on the thin cuttings (0,7 m), for 25 the silicon detection in samples. This method gives the constitutive elements identification of the biologic tissue 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 30 "electronic sonding" with an energy between 0,1 to 50 Kev. The X ray spectra emitted 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, all elements with an atomic 35 number upper than 4 are detected. The limit of 28 6 2 1 3 detection is around 10"14 to 10~15 g, which corresponds 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 5 localize the compartment of the sample in which silicon is. Because the secondary ionic analysis induces erosion of the cuttings, it will be preferable, for this technique, to use the semi thin cuttings (2 m) which results from the osmium tetraoxide post fixation. 10 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 15 function of their ration between mass and charge (m/z). 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 20 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 25 with bi-directional focusing gives a secondary ionic image filtered of the solid sample surface. The specificity of this method is that it gives an identification of all of the elements of the periodic classification and an image of the distribution with a spatial resolution under one 30 micrometer. Unfortunately, the spectrometric mass analyses induce artefacts and can be confounded between silicon and compounds with the .same mass 28. This compound could be only A1-H+ because the others, like C2H4, N2, CO,CNH2, are 9 286213 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 5 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, that means that, the located (during the micro analyse) part of sample really contains silicon. 10 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), 15 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 branches. 20 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. 25 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 species and is well indicated for Bonnemaisoniaceae 30 family, and especially for Asparagopsis Armata specie.As a matter of fact, some algae like Asparagopsis Armata have harpoons branches : each Asparagopsis plants have 5 to 10 harpoons branches such as the branch 10 with its harpoons 12 on the picture 1. In the natural environment, branches 10 2-8 6 2 1 3 away by the stream and fix themselves on new support to induce, by this way, species propagation and perenniality. to cut every harpoon branches (with 10 to 20 harpoons by 5 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 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 15 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. The support 24 which is fit up in the culture field by the moving boat is choose preferentially shaggy and smoothless to fix microslips 20 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 25 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 30 which possess a free mobility around its axe. The second one (32) is an empty plastic tube setting freely on a tight 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. 35 This limited contact is necessary to avoid The culture process uses the microslip which consists 10 very 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 11 2 8 6 213 microslips from ropes that should certainly arrives if the directing organ (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 5 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 "Power block" type (hydraulic driving) with spur surface to reduce the contact of the 10 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 15 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 20 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 pulled by a moving boat (not shown) in the same way as the rope 24. When 25 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 30 rope 24, the spout 44 outlet is equipped with a brake 48 made of a rubber ring. The sheath 46 is meant to maintain the hooking of the microcuttings to the culture support. It should be noted that it is made of. "degradable" matter so as to be 35 decomposed slowly into seawater in 10 to 20 days, the 12 286213 sufficient time to allow the fixing of the microcuttings on the support without obstructing their growth beginning. Then the culture support or rope 24, surrounding with its sheath, is submerged in a culture field, at an optimal 5 depth of about 1 meter to allow a good penetration of daylight. The supports are maintained parallel to the 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 10 equipped with floats. Therefore such a device allows to accommodate 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 15 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 20 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 25 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 30 the biological evolution of the algae leads to produce reproductive organs yearly which will be ready for picking in July and August. EXTRACTION PROCESS 13 286213 The localization method as described previously, allows to select the algae species which are interesting for extraction of organic compounds of silicon according to the invention, as well it allows a selective extraction 5 of the organic compounds of silicon from the alga parts the most 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 10 the ultra-high pressure. The cryopulverization consists of a previous freezing of the algae by 5-6 kg sheets. These sheets are introduced into a crusher where liquid nitrogen is injected so as to obtain a compound at minus 50 degrees celsium finally. 15 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 20 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 30 mn. So every alga cell undergoes a isostatic and omnidirectional high-pressure leading to its bursting. 25 As soon as the cell walls are bursted, it is essential to precipitate the alga thickeners (i.e. the carrageenans) by ammonium -like compounds or every other process Even if the organic compounds of silicon are 30 stabilized partly ->y 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 polycondensation according to the Si-OH 35 linkages. The used stabilizers could be carl 14 286213 hydroxyacid compounds/ particularly the a- and b-hydroxyacids, the glucuronides, hydroxyl or phenol aminoacids/ compounds with many alcohol (or phenol) groups. In this class we mention the glycols, the 5 catechols and the catecholamines, the polyalcohols such as the glycerol, the monosaccharides or phenol acids such as gallic acid. After the stabilization, the concentrates of organic compounds of silicon are purified at the end by ultra-10 centrifugation (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 content of the solution obtained with this extraction 15 method was about 0.3 g/1. APPLICATIONS In general, the compounds extracted with the aforesaid method will be applied in many ways owing to the 20 fact that they have the properties of the organic silicon 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 25 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 ,4). 30 Example 1 (dietetic therapy) Capsules have been made. One capsule contains 0.59 g of organic compound of silicon extracted from algae as the invention principles in a pulverulent form (the extract 15 286213 is adsorbed on a polyamide powder) that is to say 0.1 mg of silicone per capsule. The posology was 6 to 9 capsules per day. The daily absorption of algal silicon set back in good form with 5 reeducation or disappearance of bone pain, the treatment having to be continued for several months. Example 2 (tissue regeneration activity) We have carried out a cell culture of human 10 fibroblasts in microtiter plates. 8 ml of a titrated solution of organic silicon compound from algal origin was diluted in one litter of MEM medium added with FCS 2,5 %, and distributed in the wells. Two control series were made containing 10 % FCS. Cell 15 multiplication is monitored using the Neutral Red coloration technique (Borenfreund 1984). The dye-wavelength absorbance measurement is directly proportional to the number of living cells. The cell multiplication was increased of 55 % in the presence of algal organic 20 silicon. Example 3 (moisturizing activity) We have made an aqueous gel containing 12 % of algal organic silicon obtained according to the invention. This 25 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~l (C=0) absorption band to the surface of the amide 1550 cm~l(C-N) 30 absorption band. We have measured daily this hydration degree, between the 4th arid 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 35 are the followings : tfV </div>

Claims

<div class="application article clearfix printTableText" id="claims"> 286213 16 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 5 1,37 for the control. These results reveal well the immediate effect , and moreover a time-lasting effect, of an evident biological moisturizing activity (bound water). 10 Example 4 (lipolytic activity) We have maintained in a survival state human adipocytes. We added 10 jil of a solution of algal organic silicon (containing 30 mg per litter of silicon) diluted in 100 ill of NH4HCO3 0,01 M, and completed to 3 ml with a 15 Krebs-ringer buffer. 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 20 compound at the chosen dose can stimulate basal lipolysis between 0 and 60 inn. The improvement is 134 % as compared to the control. 17 28 6 2 1 3 WHAT WE CLAIM 13

1. Biologically active organic silicon compound having as a general formula: in which Rl/ R2, R3, and R4 are organic radicals with at least one of them being hydrolysable/ of the type -X-R', 10 where X is an hetero-atom and R' being an atom of hydrogen or any organic radical; characterized in that said compound is an extract of algal origin coming essentially from the reproductive organs and the young tissues of the alga. 15

2. Compound according to claim 1 in that said hetero- atom 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 performed on a sample of the 20 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 25 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. 5

4. Method according to claim 3/ in which said 30 fixation step of the, said sample of alga in a polymerized resin includes the following steps: 18 28 6 213 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 5 increasing degree of alcohol, 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. 10

5. 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 spectrometry, carry out an ionic analysis of the secondary ions in 15 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. 20

6. Procedure for the culture of the alga from which is extracted the organic silicium-containing compound according to the claim 1 or 2, charaterized in that it includes the following steps : the algal sapling to be cultured is cut into 25 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 30 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. /> jL Vi* 28 6 2 13 19

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 5 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 which said rope is shaggy in order to allow a good fixation of the said microslips on the said rope. 10

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.

10. Culture procedure according to any of the claims 15 6 to 9, in which the said algal sapling is choosen among algae that are bearing harpoon-like twigs , 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 20 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 25 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 30 hydrolysis of of the linkages Si-OR', and purify the concentrates resulting of the stabilization prcedure by ultra-centrifugation.

13. Extraction procedure according to the claim 12, in which the burst of the cells in the parts of the alga 20 28 6 2 1 3 that contain the said organic silicon compound/ is obtained by cryopounding at about -50°C.

14. 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 ultra high pressure, comprised between 4000 and 10000 bars.

15. Application of the organic silicon compound according to the claim 1 or 2, in order to obtain a therapeutical, dietary or cosmetic composition, having an 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 biologically active organic silicon compound of the general formula defined in claim 1 substantially as herein described.

17. A method of localization of the algae parts substantially as herein described with reference to the Localization Method description.

18. A process for the culture of algae substantially as herein described with reference to die accompanying drawings.

19. A process for the extraction of a biologically active organic silicon compound fiom algae substantially as herein described with reference to the Extraction Process description.

20. Application of a biologically active oiganic silicon compound substantially as herein described with reference to any example thereof

21. Any invention described or claimed herein. y the authorised agents •. J PARK & SON Ex^moi saMj AtgUiss eJ- me/ ^Afc-L. -QOC3.— 286213 Technical field \ The present invention relates to compounds based on biologically active silicon and particularly an extraction process of biologically active silicon compounds from 5 seaweeds, 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. State of the art 10 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 15 living organisms. 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 20 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 25 can be assimilated by the body, are only observed if they form, in a solution, soluble oligomers, coming from, the linkage of 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 30 silicon : silicic acid, its formula being Si(OH)4. But knowing its strong ability to polycondensate for forming silica, this compound has to be stabilized immediately after the extraction. One of the difficulties found by the applicants has 35 been to obtain silicon based compounds in which silicon is 3 28621 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 5 extract directly these compounds from vegetals. Unfortunately, all vegetals do not contain these type of silicon base compounds. Summary of the invention 10 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 15 multiplication mechanism between human embryo tissues and sea weed origin tissue. That is why the major object of the invention is to provide biologically active organic compounds from seaweed origin and to localize precisely which part of the 20 seaweeds contained said biologically active organic silicon compounds. Another object of the invention is to carry out the extraction from seaweed of biologically active silicon organic compounds. 25 Another object of the invention is to carry out the cultivation of seaweed 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 ft 30 formula is : X R—Si-R4 R3 28 6 211 in which Rl, R2, R3 and R4 are organic radicals, with at least one of which being hydrolysable, of an -X-R' type, X being a heteroatom and R' being H or an organic 5 radical, said compound being an extract of algal origin coming essentially from the reproductive organs and the young tissues of the alga. Another purpose of the this invention is a method of localisation of the parts of the sea weeds from which may 10 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 15 analysis of one of those cuts for detection of which parts contained silicon at a concentration above a predetermined level. Another purpose of the invention is an extraction method of organic silicon compound according to the 20 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 25 hydrolysing the Si-OR' bonds, and in purifying the concentrates obtained by an ultra centrifugation stabilisation. Another purpose of the invention is a process of cultivating the sea weeds from which will be extracted the 30 organic silicon compounds in which a sapling of seaweed whose, cultivation is realized is sliced in many fragments so 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 35 so as for the microslips to hook on the cultivation 5 2862 support and at the end, the cultivation support on which are hooked the microslips is installed on a cultivation field in the sea, at a predetermined depth and parallel to the sea surface. 5 Brief description of the drawings The objects, purposes and characteristics of the present invention will be better highlighted by reading the following description, being made in reference to the 10 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 15 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, 20 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. Detailed description of the invention 25 The iapplicants have found that certain parts of macroscopic seaweeds contained biologically active organic silicon compounds of general formula : R2 I Rt—Si—R4 R3 30 in which Rl, R2, R3 and R4 are organic radicals at least one of which is hydrolysable, of an -X-R1 type, X 28621 being a heteroatom and R' being H or an organic radical. The heterotaom being preferably an oxygen atom. An original localization method, as described here under, proves that those silicon compounds are mainly 5 located in the reproductive organs and or the young tissues of certain sea weed species. LOCALIZATION METHOD Generally sea weeds contain silicon, whether as 10 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 15 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 20 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. 25 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 according to the above described method. For one given sea weed variety, the preferred method 30 of localization consist in setting the sea weed sample in a resin, then to slice the samples fixed in thin cuttings in order to realise a tissue microanalyse which consists in X-ray spectrometric analyse followed by an ionic analyse. 7 28621 To fix samples to the resin, the first step consists in 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 5 less than 10"" bar. Fixed samples can be post fixed by Osmium 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 10 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 in cells, when the vacuum disappears. This technique induces step by step a 15 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 m) and 20 semi thin layer (2 m) 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 m), for 25 the silicon detection in samples. This method gives the constitutive elements identification of the biologic tissue 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 30 "electronic sonding" with ah energy between 0,1 to 50 Kev. The X ray spectra emitted 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, all elements with an atomic 35 number upper than 4 are detected. The limit of this 8 28621 detection is around 10~14 to 10~15 g, which corresponds 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 5 localize the compartment of the sample in which silicon is. Because the secondary ionic analysis induces erosion of the cuttings, it will be preferable, for this technique, to use the semi thin cuttings (2 m) which results from the osmium tetraoxide post fixation. 10 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 15 function of their ration between mass and charge (m/z). 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 20 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 25 with bi-directional focusing gives a secondary ionic image filtered of the solid sample surface. The specificity of this method is that it gives an identification of all of the elements of the periodic classification and an image of the distribution with a spatial resolution under one 30 micrometer. Unfortunately, the spectrometric mass analyses induce 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 35 usual constituents of the biological compounds and give a 9 2862 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 5 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, that means that,, the located (during the micro analyse) part of sample really contains silicon. 10 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), 15 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 branches. 20 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. 25 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 species and is well indicated for Bonnemaisoniaceae 30 family, and especially for Asparagopsis Armata specie.As a matter of fact, some algae like Asparagopsis Armata have harpoons branches : each Asparagopsis plants have 5 to 10 harpoons branches such as the branch 10 with its harpoons 12 on the picture 1. In the natural environment, branches 35 which come off by the sea mechanical action, are carried 10 286213 away by the stream 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 5 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 difficult to avoid) it is possible to obtain a potential plant multiplication up to 50 to 200 times. 10 One of the essential characteristic of this* pirocess 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 15 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. The support 24 which is fit up in the culture field by the moving boat is choose preferentially shaggy and smoothless to fix microslips 20 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 25 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 30 which possess a free mobility around its axe. The second one (32) is an empty plastic tube setting freely on a tight 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. 35 This limited contact is necessary to avoid taking off 11 2862 microslips from ropes that should certainly arrives if the directing organ (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 5 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 "Power block" type (hydraulic driving) with spur surface to reduce the contact of the 10 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 15 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 20 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 pulled by a moving boat (not shown) in the same way as the rope 24. When 25 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 30 rope 24, the spout 44 outlet is equipped with a brake 48 made of a rubber ring. The sheath 46 is meant to maintain the hooking of the microcuttings to the culture support. It should be noted that it is made of "degradable" matter so as to be 35 decomposed slowly into seawater in 10 to 20 days, the 12 2862 sufficient time to allow the fixing of the microcuttings on the support without obstructing their growth beginning. Then the culture support or rope 24, surrounding with its sheath, is submerged in a culture field, at an optimal 5 depth of about 1 meter to allow a good penetration of daylight. The supports are maintained parallel to the 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 10 equipped with floats. Therefore such a device allows to accommodate 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 15 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 20 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 25 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 30 the biological evolution of the algae leads to produce reproductive organs yearly which will be ready for picking in July and August. EXTRACTION PROCESS 13 2862 The localization method as described - previously, allows to select the algae species which are interesting for extraction of organic compounds of silicon according to the invention, as well it allows a selective extraction 5 of the organic compounds of silicon from the alga parts the most interesting according to thir ext>action. To realize the extraction of the oryai ic compounds of silicon, it is essential to burst th* • **xlular walls of the alga. 2 methods can be used : the cryopulverization or 10 the ultra-high pressure. The cryopulverization consists of a previous freezing of the algae by 5-6 kg sheets. These sheets are introduced into a crusher where liquid nitrogen is injected so as to obtain a compound at minus 50 degrees Celsium finally. 15 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 20 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 30 mn. So every alga cell undergoes a isostatic and omnidirectional high-pressure leading to its bursting. 25 As soon as the cell walls are bursted, it is essential to precipitate the alga thickeners (i.e. the carrageenans) by ammonium -like compounds or every other process Even if the organic compounds of silicon are 30 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 polycondensation according to the Si-OH 35 linkages. The used stabilizers could be carboxyl £86213 hydroxyacid compounds, particularly the fe- and - b-hydroxyacids, the glucuronides, hydroxyl or phenol aminoacids, compounds with many alcohol (or phenol) groups. In this class we mention the glycols, the 5 catechols and the catecholamines, the polyalcohols such as the glycerol, the monosaccharides or phenol acids such as gallic acid. After the stabilization, the concentrates of organic compounds of silicon are purified at the end by ultra-10 centrifugation (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 content of the solution obtained with this extraction 15 method was about 0,3 g/1. APPLICATIONS In general, the compounds extracted with the aforesaid method will be applied in many ways owing to the 20 fact that they have the properties of the organic silicon 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 25 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 ,4). 30 Example 1 (dietetic therapy) Capsules have been made. One capsule contains 0.59 g of organic compound of silicon extracted from algae as the invention principles in a pulverulent form (the extract 2862 is adsorbed on a polyamide powder) that is say 0.1. mg of silicone per capsule. The posology was 6 to 9 capsules per day. The daily absorption of algal silicon set back in good form with 5 reeducation or disappearance of bone pain, the treatment having to be continued for several months. Example 2 (tissue regeneration activity) We have carried out a cell culture of human 10 fibroblasts in microtiter plates. 8 ml of a titrated solution of organic silicon compound from algal origin was diluted in one litter of MEM medium added with FCS 2,5 %, and distributed in the wells. Two control series were made containing 10 % FCS. Cell 15 multiplication is monitored using the Neutral Red coloration technique (Borenfreund 1984) . The dye-wavelength absorbance measurement is directly proportional to the number of living cells. The cell multiplication was increased of 55 % in the presence of algal organic 20 silicon. Example 3 (moisturizing activity) We have made an aqueous gel containing 12 % of algal organic silicon obtained according to the invention. This 25 rrel 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 (C^O) absorption band to the surface of the amide 1550 cm"1(C-N) 30 absorption band. We have measured daily this 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 cut on six persons. The mean values 35 are the followings : 16 2862 73 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 5 1,37 for the control. These results reveal well the immediate effect , and moreover a time-lasting effect, of an evident biological moisturizing activity (bound water). 10 Example 4 (lipolytic activity) We have maintained in a survival state human adipocytes. We added 10 pi of a solution of algal organic silicon (containing 30 mg per litter of silicon) diluted in 100 ]il of NH4HCO3 0,01 M, and completed to 3 ml with a 15 Krebs-ringer buffer. 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 20 compound at the chosen dose can stimulate basal lipolysis between 0 and 60 mn. The improvement is 134 % as compared to the control.

1. Biologically active organic silicon compound having as a general formula: * R!—Sji-R, R3 in which Rl, R2, R3, and R4 are organic radicals with at least one of them being hydrolysable, of the type -X-R', 10 where X is an hetero-atom and R1 being an atom of hydrogen or any organic radical; characterized in that said compound is an extract of algal origin coming substantially from the reproductive organs and the young tissues of the alga. 15 2. Compound according to claim 1 in that said hetero- atom 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 performed on a sample of the 20 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 25 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 30 fixation step of the said sample of alga in a polymerized resin includes the following steps: 18 8 62 1 3 fixation of the said sample by th< fixative such as glutaraldehyde, and under a slight^ac dehydration of the said fixed sample by successive bathing in a series of alcohol-containing bath of 5 increasing degree of alcohol, 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. 10 5. 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 spectrometry, carry out an ionic analysis of the secondary ions in 15 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. 20 6. Procedure for the culture of the alga from which is extracted the organic silicon -.-containing compound according to the claim 1 or 2, charaterized in that it includes the following steps : the algal sapling to be cultured is cut into 25 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 30 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. 286213

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 5 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 which said rope is shaggy in order to allow a good fixation of the said microslips on the said rope. 10 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.

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 dbmp< obtained by cryopounding at about -50°C.

14. Extraction procedure according to the Cu-axm a*, 2 8 6 2 13;* in which the burst of the cells in the parts of the alga that contain the said organic silicon compound , is obtained by ultra high pressure, comprised between 4000 and 10000 bars.

18. A process for the culture of algae substantially as herein described with reference to the accompanying drawings.

20. Application of a biologically active organic silicon compound substantially as herein described with reference to any example thereof. Ri_CL,u£^ GT <\A£<^ rt.fc.-t- By the authorised agents Oc-G^i AJ RARK & SON 1/2 286213 <N d LL • • 2/2 2862j3 fig. 1 -34 fig. 3 fig. 4 </div>